WO2024100919A1 - Agent containing sphingoid base for use in prevention of pruritus, and use thereof - Google Patents

Agent containing sphingoid base for use in prevention of pruritus, and use thereof Download PDF

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WO2024100919A1
WO2024100919A1 PCT/JP2023/022157 JP2023022157W WO2024100919A1 WO 2024100919 A1 WO2024100919 A1 WO 2024100919A1 JP 2023022157 W JP2023022157 W JP 2023022157W WO 2024100919 A1 WO2024100919 A1 WO 2024100919A1
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gene
acid
sphingosine
expression
agent
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PCT/JP2023/022157
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French (fr)
Japanese (ja)
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達治 高橋
知也 岡本
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一丸ファルコス株式会社
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Publication of WO2024100919A1 publication Critical patent/WO2024100919A1/en

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  • the present disclosure relates to an agent containing a sphingoid base for use in suppressing pruritus and its use.
  • the stratum corneum the outermost layer of the skin, has a barrier function that prevents evaporation of moisture from inside the skin and protects against the intrusion of foreign substances such as allergens and bacteria from the outside world.
  • the stratum corneum In normal skin, the stratum corneum has intercellular lipids in the gaps between the keratinocytes, with ceramide making up approximately 50% of the intercellular lipids.
  • a decrease in ceramide correlates with atopic dermatitis, dry winter skin, aging, and the like, and causes a decline in the barrier function. For this reason, various cosmetics and topical agents that focus on ceramide have been investigated with the aim of improving the skin's barrier function (Patent Document 1).
  • the present disclosure therefore aims to provide a new agent that can, for example, induce the effect of suppressing the expression of pruritus-related genes and that can be applied primarily to human skin, etc.
  • the agent for use in suppressing pruritus of the present disclosure contains a sphingoid base.
  • TARC gene expression inhibitor contains a sphingoid base.
  • the agent disclosed herein for use in inhibiting the expression of the thymic stromal lymphopoietin (TSLP) gene contains a sphingoid base.
  • the agent disclosed herein for use in suppressing expression of the IL-31RA gene contains a sphingoid base.
  • the present disclosure provides a new agent that can induce the inhibitory effect of pruritus-related genes and can be applied primarily to human skin, etc.
  • FIG. 1 is a graph showing the amount of TARC produced in HaCaT cells in Example 1.
  • FIG. 2 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 1.
  • FIG. 3 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 1.
  • FIG. 4 is a graph showing the amount of TSLP produced in NHEK cells in Example 1.
  • FIG. 5 is a graph showing the expression level of IL-31RA gene in NHEK cells in Example 1.
  • FIG. 6 is a graph showing the amount of LTB4 produced in NHEK cells in Example 1.
  • FIG. 7 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 2.
  • FIG. 8 is a graph showing the expression level of IL-31RA gene in NHEK cells in Example 2.
  • FIG. 9 is a graph showing the expression level of IL-8 gene in NHEK cells in Example 2.
  • the present disclosure provides an agent or composition for use in suppressing pruritus.
  • the agent for use in suppressing pruritus of the present disclosure includes a sphingoid base.
  • the composition for use in suppressing pruritus of the present disclosure includes a sphingoid base.
  • sphingoid bases which are components of ceramide, exhibit an inhibitory effect on the expression of genes associated with pruritus (pruritus-related genes, biomarker genes) in keratinocytes, and have established the present disclosure.
  • the agent or composition for use in suppressing pruritus of the present disclosure is expected to be able to suppress pruritus in the skin, for example, by suppressing the expression of genes that have been shown to be associated with pruritus.
  • the term "sphingoid base” refers to a long-chain alcohol having an amino group, i.e., a long-chain amino alcohol.
  • the sphingoid base may be derived from, for example, an animal or a plant.
  • the sphingoid base may be represented, for example, by the following formula (1).
  • R 1 is a saturated or unsaturated hydrocarbon group.
  • Examples of the sphingoid base include sphingosine, sphinganine, sphingadienine, sphingenine, and the like.
  • Examples of the sphingadienine include isomers.
  • Examples of the sphingadienine isomers include 4-8-sphingadienine, in which the double bonds at the 4- and 8-positions are cis:cis, cis:trans, trans:cis, or trans:trans.
  • Examples of the sphingoid base include 4-hydroxy-8-sphingenin. The number of carbon atoms in the sphingoid base is, for example, 12 to 20, and more preferably 18.
  • the sphingoid base include animal-derived sphingoid bases such as C18-sphingosine (d18:1), Sph3), sphinganine (d18:0), and phytosphingosine (t18:0); trans-4-cis-8-sphingadienine (d18:2), Sph1, 4-hydroxy-cis-8-sphingenine (4-Hyd
  • sphingoid bases derived from plants include C20-sphingosine (C20-sphingosine, (d20:1), Sph4), and the like; preferably, 4-8-sphingadienine is used because it can more strongly suppress the expression of the biomarker gene, and more preferably, trans-4-cis-8-sphingadienine is used.
  • the sphingoid base may be, for example, a component capable of releasing a sphingoid base by being decomposed by an enzyme or the like in a living body.
  • components capable of releasing a sphingoid base include ceramide and sphingolipids.
  • the ceramide is decomposed by ceramidase in a living body to release the sphingoid base.
  • the sphingolipid is decomposed by sphingomyelinase or glycosylceramidase in a living body to release ceramide, and the ceramide is further decomposed by the ceramidase to release the sphingoid base.
  • the ceramide and the sphingolipid can be referred to as, for example, a precursor, precursor substance, or prodrug of the sphingoid base.
  • the "ceramide” refers to a compound that is composed of a sphingoid base portion and a fatty acid portion, and in which the sphingoid base portion and the fatty acid portion are bound to each other.
  • the sphingoid base portion and the fatty acid portion are covalently bound to each other by an amide bond, as shown in the following formula (6).
  • the ceramide can be represented, for example, by the following formula (6).
  • the sphingoid base portion refers to a portion of the ceramide that is derived from a sphingoid base.
  • the sphingoid base portion can be, for example, a group in which a hydrogen atom of an amino group (NH 2 ) has been eliminated.
  • the fatty acid portion refers to a portion of the ceramide that is derived from a fatty acid.
  • R 2 is, for example, a saturated or unsaturated linear hydrocarbon group, a saturated or unsaturated linear hydrocarbon group having an ⁇ -hydroxylated end, or a saturated or unsaturated linear hydrocarbon group having an ester ⁇ -hydroxylated end.
  • the number of carbon atoms in the fatty acid moiety is, for example, 16 to 24.
  • the fatty acid moiety is derived from, for example, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), and nervonic acid (24:1).
  • the fatty acid moiety is, for example, a group obtained by eliminating a hydroxyl group (OH) from the examples of the fatty acids.
  • the ceramide is, for example, N-acylsphingosine (ceramide), N-acyldihydrosphingosine (dihydroceramide), and N-acylphytosphingosine (phytoceramide).
  • the ceramide is preferably a ceramide in which the sphingoid base moiety is derived from 4-8-sphingadienine, and more preferably a ceramide in which the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, because this can more strongly suppress the expression of the biomarker gene, for example.
  • the sphingolipid means a lipid (complex lipid) in which the ceramide is further bound to a group containing a sugar or phosphoric acid.
  • the sphingolipid is a lipid in which the ceramide is bound to a sugar
  • the primary alcohol of the ceramide and the group containing the sugar are covalently bound by a glycoside bond.
  • the sphingolipid is a lipid in which the ceramide is bound to a group containing a phosphoric acid
  • the primary alcohol of the ceramide and the group containing the phosphoric acid are covalently bound by an ester bond as shown in the following formula (7).
  • the sphingolipid include sphingoglycolipids and sphingophospholipids.
  • the sphingolipid can be represented by the following formula (7), for example.
  • the sphingolipid when R 3 is a group derived from a sugar, the sphingolipid can be called a sphingoglycolipid.
  • the sphingolipid when R 3 is a group containing a phosphoric acid, the sphingolipid can be called a sphingophospholipid.
  • the sphingoglycolipid can be, for example, a compound in which the ceramide further has a sugar moiety.
  • the sugar moiety refers to the portion of the sphingoglycolipid that is derived from a sugar. Examples of the sugar include glucose, galactose, and lactose.
  • glucose glucose
  • galactose lactose
  • lactosylceramide lactosylceramide
  • the sugar moiety is preferably derived from glucose.
  • the sphingophospholipid can also be said to be a compound in which a phosphate compound is ester-bonded to the ceramide.
  • An example of the phosphate compound is phosphocholine.
  • the sphingophospholipid is called sphingomyelin.
  • the sphingoid base, the ceramide, or the sphingolipid may be an isolated or purified compound, or a composition containing a sphingoid base component.
  • the sphingoid base component may be, for example, self-prepared or a commercially available product.
  • trans-4-cis-8-sphingadienine (Nagara Science Co., Ltd.), 4-Hydroxy-cis-8-sphingenin (Nagara Science Co., Ltd.), C18-sphingosine (Nagara Science Co., Ltd.), C20-sphingosine (Nagara Science Co., Ltd.), etc.
  • a commercially available product is used as the ceramide
  • natural plant glucosylceramide Glucosylceramide, Nagara Science Co., Ltd. (URL: https://www.funakoshi.co.jp/contents/3459)
  • Glucosylceramide Nagara Science Co., Ltd. (URL: https://www.funakoshi.co.jp/contents/3459)
  • sphingomyelin When using a commercially available product as the sphingolipid, sphingomyelin (Orbract Surdary Research Laboratories (URL: https://www.funakoshi.co.jp/contents/3311)) can be used.
  • the composition containing the sphingoid base component include an extract containing the sphingoid base component; a crude product of the extract, a dried product of the extract, a freeze-dried product of the extract, a spray-dried product, and the like.
  • the extract containing the sphingoid base component can be produced, for example, by performing a solvent extraction on a plant containing the sphingoid base component.
  • the plant containing the sphingoid base include rice, corn, konnyaku, soybeans, wheat, and yeast.
  • the plant containing the ceramide include rice, corn, konnyaku, soybeans, wheat, and yeast.
  • the plant containing the sphingolipid include rice, corn, konnyaku, soybeans, wheat, and yeast.
  • One type of plant may be used, or two or more types may be used.
  • the plant material to be subjected to the extraction may be an individual plant or a part of a plant.
  • the part of the plant examples include roots, rhizomes, leaves, stems, whole flowers, or a mixture thereof.
  • the material may be the plant itself as collected, or a processed product that has been dried and/or crushed.
  • the extract containing the sphingoid base may further be contacted with ceramidase to produce the sphingoid base from the ceramide or the sphingolipid in the extract, for example.
  • the solvent used to extract the sphingoid base components is, for example, an aqueous solvent such as water or a buffer solution; a lower alcohol or a water-containing lower alcohol such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, or isobutanol; propylene glycol, 1,3-butylene glycol, 1,2-butylene glycol, 1,4-butylene glycol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, 1,3,5-pentanetriol, glycol
  • the solvent include polyhydric alcohols or hydrous polyhydric alcohols such as serine and polyethylene glycol (e.g., molecular weight 100 to 100,000); organic solvents such as acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, diox
  • the treatment of the treated material includes, for example, decomposition by adding acid (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, organic acid, etc.) or alkali (sodium hydroxide, calcium hydroxide, ammonia, etc.); fermentation or metabolic conversion by microorganisms; component adsorption by ion exchange resins, activated carbon, diatomaceous earth, etc.; fractionation using chromatography with various separation modes (ion exchange, hydrophilic adsorption, hydrophobic adsorption, size exclusion, ligand exchange, affinity, etc.); filtration using filter paper, membrane filters, ultrafiltration membranes, etc.; pressurization or decompression; heating or cooling; drying or freeze-drying; pH adjustment; deodorization; decolorization; long-term static storage; etc.
  • the above treatments may be performed alone or in combination of two or more.
  • pruritus refers to itching.
  • skin diseases accompanied by pruritus include dry skin, atopic dermatitis, contact dermatitis, and fungal skin infections.
  • the "suppression of itch” means that itch is suppressed.
  • the suppression of itch can be evaluated, for example, directly or indirectly.
  • the suppression of itch can be evaluated, for example, by the cessation of aggravation, worsening, or intensification of itch, or the improvement of itch, in a subject administered with the subject, using a model animal (e.g., an atopic dermatitis model) or a patch test.
  • the suppression of itch can be evaluated, for example, by measuring the degree of expression of itch-related genes, specifically, by the suppression of expression of the itch-related genes.
  • the pruritus-related gene refers to a gene that is related to the induction of pruritus or the formation of a skin condition that is prone to pruritus.
  • Examples of the pruritus-related gene include the TARC gene, the TSLP gene, and the IL-31RA gene.
  • TARC Thymus and Activation-Regulated Chemokine
  • TARC gene is the mRNA encoded by the human TARC gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_002987.3.
  • the expression level of the TARC gene the expression level of any one or more isoforms of the TARC gene may be measured, or the expression of all isoforms may be measured.
  • TSLP thymic stromal lymphopoietin
  • TSLP gene is the mRNA encoded by the human TSLP gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_033035.5.
  • the expression level of the TSLP gene the expression level of any one or more isoforms of the TSLP gene may be measured, or the expression of all isoforms may be measured.
  • IL-31RA Interleukin-31 receptor
  • T cell cytokine IL-31 T cell cytokine IL-31, which is secreted from Th2 cells and the like. It is known that when IL-31 binds to IL-31RA, it induces pruritus.
  • IL-31RA gene is the mRNA encoded by the human IL-31RA gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_001242636.2.
  • the expression level of the IL-31RA gene the expression level of any one or more isoforms of the IL-31RA gene may be measured, or the expression of all isoforms may be measured.
  • IL-8 Interleukin-8
  • IL-8 is a type of cytokine (chemokine) that induces neutrophil migration.
  • IL-8 is known to be the cause of inflammatory reactions.
  • IL-8 is also known as CXCL8.
  • IL-8 gene is the mRNA encoded by the human IL-8 gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_001354840.3.
  • the expression level of the IL-8 gene the expression level of any one or more isoforms of the IL-8 gene may be measured, or the expression of all isoforms may be measured.
  • suppression of gene expression means that the expression level of a target gene is suppressed or reduced, and may mean that the expression of the target gene changes from a state in which it is expressed to a state in which it is not expressed.
  • the expression of the target gene can be evaluated, for example, by measuring the expression level of the mRNA of the target gene by quantitative PCR in accordance with Examples 1 or 2 described below.
  • the expression level of the target gene may be measured by measuring the expression level of one or more isoforms of the target gene, or the expression of all isoforms, with the latter being preferred.
  • the “gene” may exist in the form of RNA (e.g., mRNA) or DNA (e.g., cDNA or genomic DNA).
  • the DNA may be double-stranded or single-stranded.
  • the “gene” may include additional sequences, such as sequences of untranslated regions (UTRs).
  • keratinocytes refers to cells that have the ability to differentiate into basal cells, spinous cells, granular cells, and keratinocytes that make up the epidermis.
  • the pruritus inhibitor of the present disclosure can, for example, inhibit the production of leukotriene B4 from keratinocytes.
  • the inhibition of leukotriene B4 production can be evaluated, for example, in accordance with Example 1 (6) described below.
  • the pruritus inhibitor of the present disclosure can inhibit pruritus, for example, by inhibiting the production of leukotriene B4.
  • the antipruritic agent of the present disclosure can suppress pruritus, for example, by administering it to a subject.
  • the conditions for use (administration conditions) of the antipruritic agent of the present disclosure are not particularly limited, and the administration form, administration timing, dosage, etc. can be appropriately set depending on, for example, the type of subject to be administered.
  • the antipruritic agent of the present disclosure may be used, for example, in vivo or in vitro .
  • the subject of administration of the pruritus suppressor of the present disclosure is not particularly limited.
  • the subject of administration can be, for example, a human or a non-human animal other than a human.
  • the non-human animal can be, for example, a mammal such as a mouse, a rat, a rabbit, a dog, a sheep, a horse, a cat, a goat, a monkey, or a guinea pig, or a bird.
  • the subject of administration can be, for example, a cell, a tissue, or an organ
  • the cell can be, for example, a cell collected from a living body, a cultured cell, or the like
  • the tissue or organ can be, for example, a tissue (living tissue) or an organ collected from a living body.
  • the amount of the sphingoid base may be within a range that exhibits an antipruritus effect, i.e., an effective amount.
  • the amount of the sphingoid base (upper and lower limits) is, for example, as follows. In the following examples, the upper and lower limits can be combined in any combination. Lower limit: 0.1 ⁇ g/ml Upper limit: 5 ⁇ g/ml, preferably 1 ⁇ g/ml
  • the administration form of the antipruritic agent of the present disclosure may be oral or parenteral.
  • parenteral administration include transdermal administration and application (contact) to the skin.
  • the application to the skin may include application to the oral mucosa, i.e., application to or contact with epithelial cells in the oral cavity.
  • the application to the skin may include administration or injection into the skin or subcutaneously via the skin surface, in addition to or instead of application to the skin surface.
  • the administration or injection into the skin via the skin surface may be performed, for example, using a microneedle.
  • the dosage form of the antipruritic agent of the present disclosure is not particularly limited and can be appropriately determined depending on, for example, the administration form.
  • Examples of the dosage form include liquid and solid forms.
  • examples of the dosage form include tablets, pills, capsules, granules, powders, liquids, etc.
  • the antipruritic agent of the present disclosure may contain, for example, additives as necessary.
  • the additives When used as a composition, the additives preferably contain pharma- ceutically acceptable additives or pharma-ceutically acceptable carriers.
  • the additives are not particularly limited, and examples include base raw materials, excipients, colorants, lubricants, binders, disintegrants, stabilizers, coating agents, preservatives, flavoring agents such as fragrances, etc.
  • the amount of the additives to be added is not particularly limited as long as they do not interfere with the function of sphingoid bases or ceramides, etc.
  • excipients examples include sugar derivatives such as lactose, lactose hydrate, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, ⁇ -starch, and dextrin; cellulose derivatives such as crystalline cellulose; organic excipients such as gum arabic, dextran, and pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, and magnesium aluminometasilicate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; and sulfates such as calcium sulfate.
  • the colorants include yellow ferric oxide.
  • Examples of the lubricants include metal stearates such as stearic acid, calcium stearate, and magnesium stearate; talc; polyethylene glycol; silica; and hydrogenated vegetable oil.
  • Examples of the flavoring agents include flavorings such as cocoa powder, peppermint, aromatic powder, peppermint oil, borneol, and cinnamon powder, as well as sweeteners and acidulants.
  • Examples of the binder include hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone, macrogol, etc.
  • disintegrant examples include cellulose derivatives such as carboxymethyl cellulose and calcium carboxymethyl cellulose; chemically modified starches and chemically modified celluloses such as carboxymethyl starch, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, and sodium starch glycolate.
  • stabilizer examples include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; sorbic acid, etc.
  • the coating agent examples include macrogols such as hypromellose and macrogol 6000, talc, titanium oxide, etc.
  • the anti-pruritus composition of the present disclosure is an orally administered composition
  • specific examples of the orally administered composition include beverages, foods, medicines (or similar), quasi-drugs (or similar), etc.
  • topical skin preparation When the pruritus suppressant or composition disclosed herein is used for transdermal administration or application to the skin (hereinafter also referred to as "topical skin preparation"), the form of the topical skin preparation may be an ampoule, capsule, powder, granule, liquid, gel, foam, emulsion, sheet, mist, spray, etc., depending on the form of use.
  • the form of use include pharmaceutical product(s); quasi-drug product(s); topical skin preparations for local or whole body use; medicinal and/or cosmetic preparations applied to the scalp and hair; bath additives for use in bath water; other preparations; etc.
  • topical or systemic skin preparations examples include basic cosmetics such as lotions, milky lotions, creams, ointments, lotions, oils and packs; face washes or skin cleansers such as solid soap, liquid soap and hand wash; massage preparations, cleansing preparations, hair removal preparations, depilatories, shaving treatments, aftershave lotions, pre-shave lotions, shaving creams, makeup cosmetics such as foundations, lipsticks, blushers, eye shadows, eyeliners and mascaras; perfumes, nail beautifying preparations, nail beautifying enamel, nail beautifying enamel removers, poultices, plasters, tapes, sheets, patches, aerosols, toothpastes and mouthwashes.
  • basic cosmetics such as lotions, milky lotions, creams, ointments, lotions, oils and packs
  • face washes or skin cleansers such as solid soap, liquid soap and hand wash
  • massage preparations cleansing preparations, hair removal preparations, depilatories, shaving treatments, aftershave lotions
  • Examples of medicinal and/or cosmetic preparations to be applied to the scalp and hair include shampoos, rinses, hair treatments, pre-hair treatments, permanent solutions, hair dyes, hair styling products, hair tonics, hair growth and care products, poultices, plasters, tapes, sheets, aerosols, etc.
  • Examples of other preparations include underarm odor prevention or deodorants, antiperspirants, sanitary products, sanitary cotton, wet tissues, etc.
  • the topical skin preparation can be produced by arbitrarily selecting and/or combining the following components and/or additives as necessary, provided that the antipruritic effect is not impaired.
  • Waxes beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax, etc.
  • Mineral oils liquid paraffin, vaseline, paraffin, ozokerite, ceresin, microcrystalline wax, etc.
  • Natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acid; and synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid, and isopentanoic acid.
  • Alcohols Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, and phenoxyethanol; and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol.
  • Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, and the like.
  • Gums, sugars or water-soluble polymer compounds Gum arabic, gum benzoin, gum dammar, guaiac butter, Irish moss, gum karaya, gum tragacanth, carob gum, quince seed, agar, casein, lactose, fructose, sucrose or esters thereof, trehalose or derivatives thereof, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C2-C4) chitin or chitosan having an alkylene (C2-C4) oxide such as ethylene oxide added thereto, low molecular weight chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or a salt thereof, Hyaluronic acid or a salt thereof, chondroitin sulf
  • Surfactants Anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkylamine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylate type amphoteric surfactants (amino type, betaine type), sulfate type amphoteric surfactants, sulfonic acid type amphoteric surfactants, phosphate type amphoteric surfactants, nonionic surfactants (ether type nonionic surfactants, ether ester type nonionic surfactants, ester type nonionic surfactants, block polymer type nonionic surfactants, nitrogen-containing type nonionic surfactants), other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, surfactants containing titanium/silicon, fluorocarbon surfactants), etc.
  • Vitamin A group retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), Vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, inositol, Vitamin C group: vitamin C acid or its derivatives, Vitamin D group: ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotachysterol, vitamin E group: vitamin E or its derivatives, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadi
  • Various Amino Acids valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine, and the like, as well as amino acid derivatives thereof, such as sulfates, phosphates, nitrates, citrates, or pyrrolidone carboxylic acid.
  • the skin topical preparation may further contain various additives derived from animals or plants.
  • the additives may be added by, for example, performing processing that is conventionally performed according to the type and form of the product to which they are to be added, and may be selected from various materials.
  • the processing may be, for example, any of the following processes selected and/or combined: crushing, milling, washing, hydrolysis, fermentation, purification, squeezing, extraction, fractionation, filtration, drying, powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc.
  • the solvent used for the extraction can be selected taking into consideration the purpose and type of the product to be used, or the subsequent processing.
  • the extraction solvent is preferably one or a mixture of two or more selected from water, lower alcohols or water-containing lower alcohols such as water, methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, and isobutanol, polyhydric alcohols or water-containing polyhydric alcohols such as propylene glycol, 1,3-butylene glycol, and glycerin, and various organic solvents such as acetone and ethyl acetate.
  • the extraction solvent may be water alone or ethanol, which is easy to remove after extraction, may be used alone or in any mixture with water, or may be extracted by squeezing.
  • the external preparations for the skin can be expected to have, for example, cosmetic effects such as protection of the skin and hair, moisturizing, improving feel and texture, imparting softness, reducing irritation, relieving stress through fragrance, activating cells (preventing cell aging), suppressing inflammation, improving skin and hair quality, preventing and improving rough skin, promoting hair growth, hair care, preventing hair loss, imparting shine, cleansing effects, relieving fatigue, promoting blood flow, and providing a warm bath effect, as well as fragrance, deodorizing, thickening, preserving, and buffering effects.
  • cosmetic effects such as protection of the skin and hair, moisturizing, improving feel and texture, imparting softness, reducing irritation, relieving stress through fragrance, activating cells (preventing cell aging), suppressing inflammation, improving skin and hair quality, preventing and improving rough skin, promoting hair growth, hair care, preventing hair loss, imparting shine, cleansing effects, relieving fatigue, promoting blood flow, and providing a warm bath effect, as well as fragrance, deodorizing, thickening, preserving, and buffering
  • the topical skin preparation may, for example, be a product that is expected to have various cosmetic and medicinal effects from the various raw materials that are known to date, and by combining these, it is possible to enhance the intended effect of this disclosure and create a product that is expected to have multifunctional effects.
  • the present disclosure provides an agent or composition that can suppress the expression of TARC gene.
  • the agent for use in suppressing the expression of TARC gene of the present disclosure comprises a sphingoid base.
  • the composition for use in suppressing the expression of TARC gene of the present disclosure comprises a sphingoid base.
  • the agent or composition for suppressing the expression of TARC gene of the present disclosure can obtain an antipruritic effect.
  • the "inhibition of expression of the TARC gene” means that the expression level of the TARC gene is inhibited or decreased, and may mean that the expression of the TARC gene changes from a state in which it is expressed to a state in which it is not expressed.
  • the expression of the TARC gene can be evaluated, for example, by measuring the expression level of the mRNA of the TARC gene according to Example 1 described below.
  • the TARC gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the TARC gene. This allows the TARC gene expression inhibitor of the present disclosure to obtain, for example, an effect of suppressing pruritus.
  • the conditions for use (administration conditions) of the TARC gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
  • the present disclosure provides an agent or composition capable of suppressing the expression of the TSLP gene.
  • the agent for use in suppressing the expression of the TSLP gene of the present disclosure comprises a sphingoid base.
  • the composition for use in suppressing the expression of the TSLP gene of the present disclosure comprises a sphingoid base.
  • the agent or composition for suppressing the expression of the TSLP gene of the present disclosure can provide an antipruritic effect.
  • the "inhibition of expression of the TSLP gene” means that the expression level of the TSLP gene is suppressed or reduced, and may mean that the expression of the TSLP gene changes from a state in which the TSLP gene is expressed to a state in which the TSLP gene is not expressed.
  • the expression of the TSLP gene can be evaluated, for example, by measuring the expression level of the mRNA of the TSLP gene in accordance with Example 1 or 2 described below.
  • the TSLP gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the TSLP gene. This allows the TSLP gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus.
  • the conditions for use (administration conditions) of the TSLP gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
  • the present disclosure provides an agent or composition capable of suppressing the expression of the IL-31RA gene.
  • the agent for use in suppressing the expression of the IL-31RA gene of the present disclosure comprises a sphingoid base.
  • the composition for use in suppressing the expression of the IL-31RA gene of the present disclosure comprises a sphingoid base.
  • the agent or composition for suppressing the expression of the IL-31RA gene of the present disclosure can provide an effect of suppressing pruritus.
  • the "suppression of expression of the IL-31RA gene” means that the expression level of the IL-31RA gene is suppressed or decreased, and may mean that the expression of the IL-31RA gene changes from a state in which it is expressed to a state in which it is not expressed.
  • the expression of the IL-31RA gene can be evaluated, for example, by measuring the expression level of the mRNA of the IL-31RA gene according to Example 1 or 2 described below.
  • the IL-31RA gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the IL-31RA gene. This allows the IL-31RA gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus.
  • the conditions for use (administration conditions) of the IL-31RA gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
  • the present disclosure provides an agent or composition capable of suppressing the expression of the IL-8 gene.
  • the agent for use in suppressing the expression of the IL-8 gene of the present disclosure comprises a sphingoid base.
  • the composition for use in suppressing the expression of the IL-8 gene of the present disclosure comprises a sphingoid base.
  • the agent or composition for suppressing the expression of the IL-8 gene of the present disclosure can provide an anti-pruritus effect.
  • the "suppression of expression of the IL-8 gene” means that the expression level of the IL-8 gene is suppressed or decreased, and may mean that the expression of the IL-8 gene changes from a state in which it is expressed to a state in which it is not expressed.
  • the expression of the IL-8 gene can be evaluated, for example, by measuring the expression level of the mRNA of the IL-8 gene according to Example 2 described below.
  • the IL-8 gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress IL-8 gene expression. This allows the IL-8 gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus.
  • the conditions for use (administration conditions) of the IL-8 gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
  • the present disclosure discloses a method capable of suppressing pruritus.
  • the method for suppressing pruritus of the present disclosure uses the agent and/or composition for use in suppressing pruritus of the present disclosure. According to the method for suppressing pruritus of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained.
  • the method for suppressing pruritus of the present disclosure includes a step of using the agent and/or composition for use in suppressing pruritus of the present disclosure on a subject.
  • the use may be, for example, contact with the skin or the like, or administration.
  • the using step may be carried out, for example, in vitro or in vivo .
  • the subject (administration subject) and administration conditions of the method for suppressing pruritus of the present disclosure the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus of the present disclosure can be cited, for example.
  • the present disclosure discloses a method for suppressing the expression of TARC gene.
  • the method for suppressing the expression of TARC gene of the present disclosure uses the expression inhibitor and/or composition of the TARC gene of the present disclosure.
  • According to the method for suppressing the expression of TARC gene of the present disclosure for example, it is expected that the effect of suppressing pruritus can be obtained by suppressing the expression of TARC gene.
  • the method for suppressing expression of the TARC gene disclosed herein includes a step of using the expression inhibitor and/or composition of the TARC gene disclosed herein on a subject.
  • the use may be, for example, contact with the skin or the like, or administration.
  • the using step may be carried out, for example, in vitro or in vivo .
  • the subject (administration subject) and administration conditions of the method for suppressing the expression of the TARC gene of the present disclosure the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus of the present disclosure can be cited, for example.
  • the present disclosure discloses a method capable of suppressing the expression of the TSLP gene.
  • the method for suppressing the expression of the TSLP gene of the present disclosure uses the expression inhibitor and/or composition of the TSLP gene of the present disclosure. According to the method for suppressing the expression of the TSLP gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing the expression of the TSLP gene.
  • the method of suppressing expression of the TSLP gene disclosed herein includes a step of using the TSLP gene expression inhibitor and/or composition disclosed herein on a subject.
  • the use may be, for example, contact with the skin or the like, or administration.
  • the using step may be carried out, for example, in vitro or in vivo .
  • the subject (administration subject) and administration conditions of the method for suppressing expression of the TSLP gene disclosed herein the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
  • the present disclosure discloses a method capable of suppressing expression of the IL-31RA gene.
  • the method for suppressing expression of the IL-31RA gene of the present disclosure uses the IL-31RA gene expression inhibitor and/or composition of the present disclosure. According to the method for suppressing expression of the IL-31RA gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing expression of the IL-31RA gene.
  • the method for suppressing expression of the IL-31RA gene disclosed herein includes a step of using the IL-31RA gene expression inhibitor and/or composition disclosed herein on a subject.
  • the use may be, for example, contact with the skin or the like, or administration.
  • the using step may be carried out, for example, in vitro or in vivo .
  • the subject (administration subject) and administration conditions of the method for suppressing expression of the IL-31RA gene disclosed herein the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
  • the present disclosure discloses a method capable of suppressing expression of the IL-8 gene.
  • the method for suppressing expression of the IL-8 gene of the present disclosure uses the IL-8 gene expression inhibitor and/or composition of the present disclosure. According to the method for suppressing expression of the IL-8 gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing expression of the IL-8 gene.
  • the method for suppressing expression of the IL-8 gene disclosed herein includes a step of using the IL-8 gene expression inhibitor and/or composition disclosed herein on a subject.
  • the use may be, for example, contact with the skin or the like, or administration.
  • the using step may be carried out, for example, in vitro or in vivo .
  • the subject (administration subject) and administration conditions of the method for suppressing expression of the IL-8 gene disclosed herein the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
  • the present disclosure relates to the use of an agent and/or composition for use in suppressing pruritus.
  • the present disclosure relates to the use of a TARC gene expression inhibitor and/or composition for use in suppressing TARC gene expression.
  • the present disclosure relates to the use of a TSLP gene expression inhibitor and/or composition for use in suppressing TSLP gene expression.
  • the present disclosure relates to the use of an IL-31RA gene expression inhibitor and/or composition for use in suppressing IL-31RA gene expression.
  • the present disclosure relates to the use of an IL-8 gene expression inhibitor and/or composition for use in suppressing IL-8 gene expression.
  • the present disclosure relates to the use of an agent and/or composition for use in suppressing pruritus, for producing an agent for use in suppressing pruritus, or a composition for use in suppressing pruritus.
  • the present disclosure relates to the use of an agent and/or composition for suppressing TARC gene expression, for producing an agent for use in suppressing TARC gene expression, or a composition for use in suppressing TARC gene expression.
  • the present disclosure relates to the use of an agent and/or composition for suppressing TSLP gene expression, for producing an agent for use in suppressing TSLP gene expression, or a composition for use in suppressing TSLP gene expression.
  • the present disclosure relates to the use of an agent and/or composition for suppressing IL-31RA gene expression, for producing an agent for use in suppressing IL-31RA gene expression, or a composition for use in suppressing IL-31RA gene expression.
  • the present disclosure relates to the use of an agent and/or composition for suppressing IL-8 gene expression, for producing an agent for use in suppressing IL-8 gene expression, or a composition for use in suppressing IL-8 gene expression.
  • TARC Production Test HaCaT cells which are human keratinocytes, were seeded on a 24-well plate.
  • the medium used was a 10% FBS-containing DMEM medium (041-29775, Fujifilm Wako Pure Chemical Industries, Ltd., the same applies below in the Examples). After the seeding, the HaCaT cells were cultured until they were 90% confluent. After the culture, the medium in each well was replaced with serum-free DMEM. After the replacement, the HaCaT cells were cultured for 24 hours.
  • Fig. 1 is a graph showing the amount of TARC produced in HaCaT cells, in which the vertical axis indicates the amount of TARC produced (pg/ml) and the horizontal axis indicates the type of sample.
  • the values on the vertical axis in FIG. 1 indicate the values for each group as the amount of TARC produced corrected by the cell number.
  • the values on the vertical axis in FIG. 1 indicate the values for each group as the amount of TARC produced corrected by the cell number.
  • the medium used was KBM (trademark) medium (calcium-phenol red-free, 00195769, manufactured by Lonza Co., Ltd., the same applies in the following examples).
  • KBM trademark
  • BPE bovine pituitary extract
  • insulin insulin
  • EGF epidermal growth factor
  • antibiotics G-1000
  • 0.06 mmol/l calcium 0.06 mmol/l calcium.
  • Fcer or Sph1 was added at a concentration of 1 ⁇ g/ml, and the cells were cultured for 1 hour.
  • the stimulatory factors TNF ⁇ and INF ⁇ were added to 10 ng/ml (final concentration) and 10 ng/ml (final concentration), respectively, and the cells were cultured for 6 hours.
  • the culture supernatant was collected, and mRNA was purified using RNeasy Mini Kit (QIAGEN) according to the attached protocol.
  • reverse transcription reaction was performed using PrimeScript Master Mix (TaKaRa).
  • qRT-PCR was performed using the following primer set for the TSLP gene and primer set for the RPS gene according to the method described in TB Green Master Mix (manufactured by TaKaRa).
  • the PCR device used was LightCycler (registered trademark) 96 (manufactured by Roche).
  • the expression level of the TSLP gene was taken as a relative value to the expression level of the internal standard gene (RPS18 gene). Data was analyzed using the accompanying software. Statistical analysis was performed using JMP8 by the Turky test and Dunnett's test. These results are shown in FIG. 2.
  • Primer set for TSLP gene Forward primer (SEQ ID NO: 1) 5'-TAGCAATCGGCCACATTGCCT-3' Reverse primer (SEQ ID NO:2) 5'-GAAGCGACGCCACAATCCTTG-3' Primer set for RPS18 gene Forward primer (SEQ ID NO: 5) 5'-TTTGCAGTACTCAACACCAACA-3' Reverse primer (SEQ ID NO:6) 5'-GAGCATATCTTCGGCCCACAC-3'
  • Fig. 2 is a graph showing the expression level of the TSLP gene in NHEK cells.
  • the vertical axis shows the relative value of the expression level of the TSLP gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of the TSLP gene, and the horizontal axis shows the type of sample.
  • the values on the vertical axis in Figures 2 (A) and (B) indicate the values for each group, with the value for the control group being 1.0 (1.00).
  • the NHEK cells were cultured for 24 hours. After the culture, 1 ⁇ g/ml of phytoceramide (Fcer) or sphingoid bases (Sph1) were added and cultured for 1 hour. After the culture, Poly(I:C) was added to 1 ⁇ g/ml, and cultured for 16 hours. After the culture, the culture supernatant was collected, and mRNA was purified using RNeasy Mini Kit (QIAGEN) according to the protocol described. After the purification, reverse transcription reaction was performed using PrimeScript Master Mix (TaKaRa).
  • qRT-PCR was performed using the primer set for the TSLP gene and the primer set for the RPS gene according to the method described in TB Green Master Mix (TaKaRa).
  • LightCycler (registered trademark) 96 (Roche) was used as the PCR device.
  • the gene expression of the TSLP gene was a relative value to the expression level of the internal standard gene (RPS18 gene).
  • centrifugation was performed after the culture supernatant was collected, and debris was removed.
  • ELISA was performed using Human TSLP Quantikine ELISA Kit (R&D System). The amount of TSLP produced was quantified from the results of the ELISA.
  • Statistical analysis was performed using the Turky test with JMP 8. The results are shown in Figures 3 and 4.
  • FIG. 3 is a graph showing the expression level of TSLP gene in NHEK cells.
  • the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
  • the values on the vertical axis in Figures 3 (A) and (B) indicate the values for each group, with the value for the control group being 1.0 (1.00).
  • Fig. 4 is a graph showing the amount of TSLP produced in NHEK cells.
  • the vertical axis shows the amount of TSLP produced, and the horizontal axis shows the type of sample.
  • Fig. 4(B) the vertical axis shows the amount of TSLP produced, and the horizontal axis shows the type of sample.
  • the values on the vertical axis in FIG. 4 indicate the values for each group as the amount of TSLP produced corrected by the cell number.
  • IL-31RA IL-31 receptor A
  • LTB4 leukotriene B4
  • Sph1 contributes to the suppression of IL-31RA gene expression in addition to the suppression of TSLP gene expression and protein production.
  • NHEK cells were seeded in a 24-well plate at 1.0 x 10 5 cells/well.
  • KBM (trademark) medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent.
  • KBM trademark
  • BPE bovine pituitary extract
  • insulin insulin
  • EGF epidermal growth factor
  • antibiotics G-1000
  • 0.06 mmol/l calcium 0.06 mmol/l calcium.
  • the NHEK cells were cultured for 24 hours.
  • 0.1 ⁇ g/ml, 0.5 ⁇ g/ml, or 1 ⁇ g/ml of phytoceramide (Fcer) or sphingoid bases (Sph1) were added and cultured for 1 hour.
  • TNF ⁇ and INF ⁇ were added to a final concentration of 10 ng/ml and cultured for 6 hours.
  • Fig. 5 is a graph showing the expression level of IL-31RA gene in NHEK cells.
  • the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the concentration of Sph1 added.
  • the numerical values on the vertical axis in Figures 5 (A) and (B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
  • NHEK cells were seeded in a 24-well plate at 1.0 ⁇ 10 5 cells/well. KBMTM medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBMTM medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotic (GA-1000), and 0.06 mM calcium. After the replacement, the NHEK cells were cultured for 24 hours.
  • sphingoid bases Sph1
  • TNF ⁇ and INF ⁇ were added to a final concentration of 10 ng/ml, and the cells were cultured for 16 hours.
  • the medium was replaced with IL-31 (100 ng/ml), BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotics (GA-1000), and 0.06 mM calcium KBMTM medium.
  • the NHEK cells were cultured for 1 hour. After the culture, the culture supernatant was collected and centrifuged to remove debris.
  • Fig. 6 is a graph showing the amount of LTB4 produced in NHEK cells, in which the vertical axis indicates the amount of LTB4 produced (pg/ml) and the horizontal axis indicates the type of sample.
  • the values on the vertical axis in FIG. 6 indicate the values for each group as the amount of LTB4 produced corrected by the cell number.
  • the amount of LTB4 production was significantly suppressed in the Sph1 group compared to the No add (negative control) group.
  • Example 2 The structural type of glucosylceramide contained in rice-derived glucosylceramide was analyzed, and the composition ratio of sphingoid base species contained in glucosylceramide was inferred from the analysis results.Furthermore, it was confirmed whether the sphingoid base contained in the rice-derived glucosylceramide exhibits an antipruritic effect.
  • glucosylceramide structure The structural type of glucosylceramide contained in rice-derived glucosylceramide was analyzed, and the type of sphingoid base contained in the rice-derived glucosylceramide was inferred. Specifically, 100 mg of rice-derived glucosylceramide (Phytocera, NW-10, Lot No. 121520, Ichimaru Falcos) was added to 3 ml of purified water, and sonicated to dissolve. After the dissolution, 18 ml of methanol was gradually added and sonicated. Then, the mixture was allowed to cool, and chloroform was added to a total volume of 50 ml to perform extraction.
  • rice-derived glucosylceramide was found to contain 81% sphingadienine (d18:2) (Sph1) and 12.6% sphingenine (t18:1) (Sph2). From the above, it was found that rice-derived glucosylceramide (Phytosera, NW-10, Lot No. 121520, Ichimaru Pharcos) mainly contains sphingadienine.
  • KBM trademark
  • BPE bovine pituitary extract
  • EGF epidermal growth factor
  • G-1000 antibiotics
  • tSph rice-derived glucosylceramide
  • the stimulatory factors TNF ⁇ and INF ⁇ were added at 1 ng/ml (final concentration) and 1 ng/ml (final concentration), respectively, and the cells were cultured for 6 hours. After the culture, qRT-PCR and analysis were performed in the same manner as in Example 1(2). The results are shown in FIG. 7.
  • FIG. 7 is a graph showing the expression level of TSLP gene in NHEK cells.
  • the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
  • the numerical values on the vertical axis in Figures 7(A) and 7(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
  • qRT-PCR and analysis were performed in the same manner as in Example 1(4) above, except that NHEK cells (KK-4009, frozen NHEK (NB), derived from a newborn, manufactured by Kurabo) were used as cells, Sph1 or rice-derived glucosylceramide (tSph) at a concentration of 0.1 ⁇ g/ml, 0.5 ⁇ g/ml, or 1 ⁇ g/ml was used as ceramide, and TNF ⁇ and INF ⁇ were used at 1 ng/ml (final concentration) as cytokines.
  • NHEK cells KK-4009, frozen NHEK (NB), derived from a newborn, manufactured by Kurabo
  • Sph1 or rice-derived glucosylceramide tSph
  • TNF ⁇ and INF ⁇ were used at 1 ng/ml (final concentration) as cytokines.
  • Fig. 8 is a graph showing the gene expression level of IL-31RA in NHEK cells.
  • the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample.
  • the numerical values on the vertical axis in FIG. 8(A) and FIG. 8(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
  • Figure 8(A) in the Sph1 group, the gene expression level of IL-31RA was significantly suppressed at all concentrations compared to the No add (negative control) group.
  • NHEK cells (KK-4009, frozen NHEK (NB), derived from a newborn, manufactured by Kurabo Industries, Ltd.) were seeded on a 24-well plate at 1.0 x 10 5 cells/well. KBM (trademark) medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent.
  • KBM trademark
  • BPE bovine pituitary extract
  • insulin insulin
  • EGF epidermal growth factor
  • antibiotic G-1000
  • 0.06 mmol/l calcium 0.06 mmol/l calcium.
  • the NHEK cells were cultured for 24 hours.
  • Sph1 at a concentration of 0.5 ⁇ g/ml or 1 ⁇ g/ml
  • rice-derived glucosylceramide at 1 ⁇ g/ml was added and cultured for 1 hour.
  • TNF ⁇ and INF ⁇ were added to a final concentration of 1 ng/ml and cultured for 6 hours.
  • Fig. 9 is a graph showing the expression level of IL-8 gene in NHEK cells.
  • the vertical axis shows the relative value of the expression level of IL-8 gene, and the horizontal axis shows the type of sample.
  • the vertical axis shows the relative value of the expression level of IL-8 gene, and the horizontal axis shows the type of sample.
  • the numerical values on the vertical axis in FIG. 9(A) and FIG. 9(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
  • ⁇ Agent containing a sphingoid base for use in suppressing pruritus> (Appendix 1) An agent for use in suppressing pruritus, comprising a sphingoid base. (Appendix 2) The agent according to claim 1, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 3 The agent described in Appendix 1, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 4 An agent for use in suppressing pruritus, comprising a ceramide having a sphingoid base portion and a fatty acid portion. (Appendix 5) 5.
  • Appendix 6 The agent according to claim 4 or 5, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition for use in suppressing pruritus containing a sphingoid base A composition for use in suppressing pruritus, comprising a sphingoid base.
  • the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • compositions for use in suppressing pruritus comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • Appendix 16 16
  • the composition of claim 15, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • composition of claim 15 or 16 wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 15 or 16 wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 19 19. The composition of any one of claims 15 to 18, wherein the ceramide further comprises a sugar moiety.
  • Appendix 20 20.
  • composition of claim 19, wherein the sugar moiety is derived from glucose is derived from glucose.
  • Appendix 21 21.
  • a composition for application to the skin comprising a composition according to any one of claims 12 to 20.
  • a composition for oral administration comprising a composition according to any one of claims 12 to 20.
  • Appendix 23 A composition described in any one of appendices 12 to 22, comprising an agent for use in suppressing pruritus described in any one of appendices 1 to 11.
  • ⁇ Agent for use in suppressing expression of TARC gene> Appendix 24
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • An agent for use in inhibiting expression of the TARC gene comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • (Appendix 28) 28 The agent of claim 27, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • (Appendix 29) The agent according to claim 27 or 28, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • An agent for oral administration comprising an agent according to any one of claims 24 to 32.
  • ⁇ Composition for use in suppressing expression of TARC gene> (Appendix 35) A composition for use in inhibiting expression of the TARC gene, comprising a sphingoid base. (Appendix 36) The composition of claim 35, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 35 wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition for use in inhibiting expression of the TARC gene comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • composition of claim 38 wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • Appendix 40 The composition of claim 38 or 39, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 38 or 39 wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 42 42.
  • the composition of any one of claims 38 to 41, wherein the ceramide further comprises a sugar moiety.
  • Appendix 43 43.
  • the composition of claim 42, wherein the sugar moiety is derived from glucose.
  • Appendix 44 44.
  • a composition for application to the skin comprising a composition according to any one of claims 35 to 43.
  • compositions for oral administration comprising a composition according to any one of claims 35 to 43.
  • Appendix 46 A composition described in any one of appendices 35 to 45, comprising an agent for use in inhibiting expression of the TARC gene described in any one of appendices 24 to 34.
  • ⁇ Agent for use in inhibiting expression of TSLP gene> An agent for use in inhibiting expression of thymic stromal lymphopoietin (TSLP) gene, comprising a sphingoid base.
  • TSLP thymic stromal lymphopoietin
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • An agent for use in inhibiting expression of the TSLP gene comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • (Appendix 51) 51 The agent of claim 50, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • (Appendix 52) 52 The agent according to claim 50 or 51, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 53 52.
  • Appendix 56 56.
  • An agent for application to the skin comprising an agent according to any one of claims 47 to 55. (Appendix 57) 56.
  • composition for use in inhibiting expression of the TSLP gene comprising a sphingoid base.
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 58 wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition for use in inhibiting expression of the TSLP gene comprising a ceramide comprising a sphingoid base moiety and a fatty acid moiety.
  • composition of claim 61 wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • (Appendix 64) The composition of claim 61 or 62, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • the composition of any one of claims 61 to 64, wherein the ceramide further comprises a sugar moiety.
  • (Appendix 67) 67 A composition for application to the skin comprising a composition according to any of claims 58 to 66. (Appendix 68) 67.
  • a composition for oral administration comprising a composition according to any one of claims 58 to 66.
  • Appendix 69 A composition described in any one of appendices 58 to 68, comprising an agent for use in inhibiting the expression of a TSLP gene described in any one of appendices 47 to 57.
  • ⁇ Agent for use in suppressing expression of IL-31RA gene> An agent for use in suppressing the expression of the IL-31RA gene, comprising a sphingoid base.
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • An agent for use in suppressing expression of the IL-31RA gene comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • An agent for oral administration comprising an agent according to any one of claims 70 to 78.
  • ⁇ Composition for use in suppressing expression of IL-31RA gene> (Appendix 81) A composition for use in suppressing expression of the IL-31RA gene, comprising a sphingoid base. (Appendix 82) The composition of claim 81, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 81 wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition for use in suppressing expression of the IL-31RA gene comprising a ceramide having a sphingoid base moiety and a fatty acid moiety.
  • Appendix 85 85.
  • composition of claim 84 wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • Appendix 86 The composition of claim 84 or 85, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • a composition for oral administration comprising a composition according to any one of claims 81 to 89.
  • Appendix 92 The composition according to any one of appendices 81 to 91, comprising an agent for use in suppressing the expression of the IL-31RA gene according to any one of appendices 70 to 80.
  • ⁇ Agent for use in suppressing expression of IL-8 gene> (Appendix 93) An agent for use in suppressing the expression of the IL-8 gene, comprising a sphingoid base.
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • An agent for use in suppressing expression of the IL-8 gene comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  • (Appendix 97) 97 The agent of claim 96, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • Appendix 98 The agent according to claim 96 or 97, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition for use in suppressing expression of IL-8 gene A composition for use in suppressing expression of the IL-8 gene, comprising a sphingoid base.
  • sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 104 wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 107 A composition for use in suppressing expression of the IL-8 gene, comprising a ceramide having a sphingoid base moiety and a fatty acid moiety.
  • Appendix 108 108.
  • composition of claim 107 wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  • the composition of claim 107 or 108, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • composition of claim 107 or 108 wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  • Appendix 111) 111.
  • the composition of any one of claims 107 to 110, wherein the ceramide further comprises a sugar moiety.
  • the composition of claim 111, wherein the sugar moiety is derived from glucose.
  • Appendix 113 A composition for application to the skin comprising the composition of any of claims 104 to 112.
  • a composition for oral administration comprising the composition of any of claims 104 to 112.
  • Appendix 115 A composition according to any one of claims 104 to 114, comprising an agent for use in suppressing expression of an IL-8 gene according to any one of claims 93 to 103.
  • Method for suppressing pruritus (Appendix 116) A method for suppressing pruritus, comprising using an agent for use in suppressing pruritus described in any one of Appendices 1 to 11 and/or a composition for use in suppressing pruritus described in any one of Appendices 12 to 23.
  • Appendix 117 The method for suppressing itch described in Appendix 116, comprising a step of using the agent for use in suppressing itch and/or the composition for use in suppressing itch on a subject.
  • Appendix 118 118.
  • ⁇ Method for inhibiting expression of TARC gene> (Appendix 119) A method for inhibiting expression of the TARC gene, comprising using an agent for inhibiting expression of the TARC gene described in any of Appendices 24 to 34 and/or a composition for inhibiting expression of the TARC gene described in any of Appendices 35 to 46.
  • Appendix 120 The method for suppressing expression of the TARC gene described in Appendix 119, comprising a step of using an agent for suppressing expression of the TARC gene and/or a composition for suppressing expression of the TARC gene in a subject.
  • Appendix 121) The method of suppression according to claim 119 or 120, for use in vitro or in vivo .
  • Appendix 122 A method for inhibiting expression of the TSLP gene, comprising using an agent for use in inhibiting expression of the TSLP gene described in any of Appendices 47 to 57, and/or a composition for use in inhibiting expression of the TSLP gene described in any of Appendices 58 to 69.
  • Appendix 123 The method of suppression described in Appendix 122, comprising a step of using an agent for use in suppressing expression of the TSLP gene and/or a composition for use in suppressing expression of the TSLP gene in a subject.
  • Appendix 124) 124.
  • Appendix 128 A method for suppressing expression of the IL-8 gene, comprising using an agent for use in suppressing expression of the IL-8 gene described in any of Appendices 93 to 103 and/or a composition for use in suppressing expression of the IL-8 gene described in any of Appendices 104 to 115.
  • Appendix 134 Use of an agent for use in suppressing expression of the IL-31RA gene according to any one of appendices 70 to 80 and/or a composition for use in suppressing expression of the IL-31RA gene according to any one of appendices 81 to 92
  • Appendix 135) Use of an agent for use in suppressing the expression of the IL-8 gene according to any one of appendices 93 to 103 and/or a composition for use in suppressing the expression of the IL-8 gene according to any one of appendices 104 to 115
  • the present disclosure can provide a new agent that can induce the effect of suppressing the expression of pruritus-related genes and can be applied primarily to human skin. For this reason, the present disclosure can be said to be extremely useful, for example, in the fields of pharmaceuticals, quasi-drugs, and topical skin preparations.

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Abstract

Provided is a novel agent which can induce an effect of inhibiting the expression of a pruritus-related gene and is mainly applicable to human skin or the like.　The agent for use in the prevention of pruritus according to the present disclosure contains a sphingoid base.

Description

スフィンゴイド塩基を含む掻痒抑制に用いるための剤およびその用途Agent for use in suppressing pruritus containing a sphingoid base and its use
　本開示は、スフィンゴイド塩基を含む掻痒抑制に用いるための剤およびその用途に関する。 The present disclosure relates to an agent containing a sphingoid base for use in suppressing pruritus and its use.
　皮膚の最外層である角質層は、皮膚内部からの水分の蒸散を防ぎ、外界からのアレルゲンや細菌等の異物の侵入を防御するバリア機能を有する。正常な皮膚において、角質層は、角質細胞の隙間に細胞間脂質を有し、細胞間脂質の約５０％をセラミドが占めている。セラミドの減少は、アトピー性皮膚炎、冬季の乾燥肌、および加齢等と相関しており、バリア機能の低下を引き起こす。このため、皮膚のバリア機能の改善を目的として、セラミドに着目した化粧料や外用剤等が種々検討されている（特許文献１）。 The stratum corneum, the outermost layer of the skin, has a barrier function that prevents evaporation of moisture from inside the skin and protects against the intrusion of foreign substances such as allergens and bacteria from the outside world. In normal skin, the stratum corneum has intercellular lipids in the gaps between the keratinocytes, with ceramide making up approximately 50% of the intercellular lipids. A decrease in ceramide correlates with atopic dermatitis, dry winter skin, aging, and the like, and causes a decline in the barrier function. For this reason, various cosmetics and topical agents that focus on ceramide have been investigated with the aim of improving the skin's barrier function (Patent Document 1).
特許第６９５９２０４号Patent No. 6959204
　そこで、本開示は、例えば、掻痒（そうよう）関連遺伝子の発現抑制効果を誘導でき、主に、ヒトの皮膚等に適用可能な新たな剤を提供すること、などを目的とする。 The present disclosure therefore aims to provide a new agent that can, for example, induce the effect of suppressing the expression of pruritus-related genes and that can be applied primarily to human skin, etc.
　前記目的を達成するために、本開示の掻痒抑制に用いるための剤（以下、「掻痒抑制剤」ともいう。）は、スフィンゴイド塩基を含む。 In order to achieve the above objective, the agent for use in suppressing pruritus of the present disclosure (hereinafter also referred to as the "pruritus suppressant") contains a sphingoid base.
　本開示のＴＡＲＣ（Thymus and Activation-Regulated Chemokine）遺伝子の発現の抑制に用いるための剤（以下、「ＴＡＲＣ遺伝子の発現抑制剤」ともいう。）は、スフィンゴイド塩基を含む。 The agent disclosed herein for use in inhibiting expression of the Thymus and Activation-Regulated Chemokine (TARC) gene (hereinafter also referred to as a "TARC gene expression inhibitor") contains a sphingoid base.
　本開示のＴＳＬＰ（Thymic stromal lymphopoietin）遺伝子の発現の抑制に用いるための剤（以下、「ＴＳＬＰ遺伝子の発現抑制剤」ともいう。）は、スフィンゴイド塩基を含む。 The agent disclosed herein for use in inhibiting the expression of the thymic stromal lymphopoietin (TSLP) gene (hereinafter also referred to as the "TSLP gene expression inhibitor") contains a sphingoid base.
　本開示のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤（以下、「ＩＬ－３１ＲＡ遺伝子の発現抑制剤」ともいう。）は、スフィンゴイド塩基を含む。 The agent disclosed herein for use in suppressing expression of the IL-31RA gene (hereinafter also referred to as the "IL-31RA gene expression suppressor") contains a sphingoid base.
　本開示によれば、掻痒関連遺伝子の抑制効果を誘導でき、主に、ヒトの皮膚等に適用可能な新たな剤を提供できる。 The present disclosure provides a new agent that can induce the inhibitory effect of pruritus-related genes and can be applied primarily to human skin, etc.
図１は、実施例１における、ＨａＣａＴ細胞における、ＴＡＲＣ産生量を示すグラフである。FIG. 1 is a graph showing the amount of TARC produced in HaCaT cells in Example 1. 図２は、実施例１における、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。FIG. 2 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 1. 図３は、実施例１における、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。FIG. 3 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 1. 図４は、実施例１における、ＮＨＥＫ細胞における、ＴＳＬＰ産生量を示すグラフである。FIG. 4 is a graph showing the amount of TSLP produced in NHEK cells in Example 1. 図５は、実施例１における、ＮＨＥＫ細胞における、ＩＬ－３１ＲＡの遺伝子発現量を示すグラフである。FIG. 5 is a graph showing the expression level of IL-31RA gene in NHEK cells in Example 1. 図６は、実施例１における、ＮＨＥＫ細胞における、ＬＴＢ４産生量を示すグラフである。FIG. 6 is a graph showing the amount of LTB4 produced in NHEK cells in Example 1. 図７は、実施例２における、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。FIG. 7 is a graph showing the expression level of the TSLP gene in NHEK cells in Example 2. 図８は、実施例２における、ＮＨＥＫ細胞における、ＩＬ－３１ＲＡの遺伝子発現量を示すグラフである。FIG. 8 is a graph showing the expression level of IL-31RA gene in NHEK cells in Example 2. 図９は、実施例２における、ＮＨＥＫ細胞における、ＩＬ－８の遺伝子発現量を示すグラフである。FIG. 9 is a graph showing the expression level of IL-8 gene in NHEK cells in Example 2.
　以下、本開示について、例をあげて具体的に説明する。以下、特に言及しない限り、各開示は、他の開示の説明を援用できる。 The present disclosure will be explained in detail below with examples. Unless otherwise specified, each disclosure may refer to the explanations of other disclosures.
＜スフィンゴイド塩基を含む掻痒抑制に用いるための剤または組成物＞
　ある態様において、本開示は、掻痒抑制に用いるための剤または組成物を提供する。本開示の掻痒抑制に用いるための剤は、スフィンゴイド塩基を含む。また、本開示の掻痒抑制に用いるための組成物は、スフィンゴイド塩基を含む。以下の説明において、特に言及しない限り、本開示の各剤の説明は、対応する各組成物の説明に援用できる。 <Agent or composition containing a sphingoid base for use in suppressing pruritus>
In one embodiment, the present disclosure provides an agent or composition for use in suppressing pruritus. The agent for use in suppressing pruritus of the present disclosure includes a sphingoid base. Also, the composition for use in suppressing pruritus of the present disclosure includes a sphingoid base. In the following description, unless otherwise specified, the description of each agent of the present disclosure can be used to refer to the description of the corresponding composition.
　本発明者らは、鋭意研究の結果、セラミドの構成部分であるスフィンゴイド塩基が、角化細胞において、掻痒と関連する遺伝子（掻痒関連遺伝子、バイオマーカー遺伝子）の発現抑制作用を示すことを見出し、本開示を確立するに至った。このため、本開示の掻痒抑制に用いるための剤または組成物によれば、例えば、掻痒との関連性が示されている遺伝子の発現を抑制できるため、皮膚における掻痒を抑制できると期待される。 As a result of intensive research, the present inventors discovered that sphingoid bases, which are components of ceramide, exhibit an inhibitory effect on the expression of genes associated with pruritus (pruritus-related genes, biomarker genes) in keratinocytes, and have established the present disclosure. For this reason, the agent or composition for use in suppressing pruritus of the present disclosure is expected to be able to suppress pruritus in the skin, for example, by suppressing the expression of genes that have been shown to be associated with pruritus.
　本開示において、「スフィンゴイド塩基」は、アミノ基を有する長鎖アルコール、すなわち、長鎖アミノアルコールを意味する。前記スフィンゴイド塩基は、例えば、動物由来でもよいし、植物由来でもよい。前記スフィンゴイド塩基は、例えば、下記式（１）で表すことができる。下記式（１）において、Ｒ１は、飽和または不飽和の炭化水素基である。前記スフィンゴイド塩基は、例えば、スフィンゴシン（Sphingosine）、スフィンガニン（Sphinganine）、スフィンガジエニン（Sphingadienine）、スフィンゲニン（sphingenin）等があげられる。前記スフィンガジエニンは、例えば、異性体があげられる。前記スフィンガジエニンの異性体は、例えば、４位および８位の二重結合が、cis:cis、cis:trans、trans:cis、またはtrans:transである、４－８－スフィンガジエニン（4-8-sphingadienine）等があげられる。前記スフィンゲニンは、例えば、４－ヒドロキシ－８－スフィンゲニン（4-Hydroxy-8-sphingenin）等があげられる。前記スフィンゴイド塩基における炭素数は、例えば、１２～２０であり、より好ましくは、１８である。具体例として、前記スフィンゴイド塩基は、例えば、Ｃ１８－スフィンゴシン（C18-sphingosine、（d18:1）、Sph3）、スフィンガニン（d18:0）、フィトスフィンゴシン（t18:0）等の動物由来のスフィンゴイド塩基；トランス－４－シス－８－スフィンガジエニン（trans-4-cis-8-sphingadienine、（d18:2）、Sph1）、４－ヒドロキシ－シス－８－スフィンゲニン（4-Hydroxy-cis-8-sphingenin、（t18:1）、Sph2）、８－スフィンゲニン（d18:1）等の前記植物由来のスフィンゴイド塩基；Ｃ２０－スフィンゴシン（C20-sphingosine、（d20:1）、Sph4）等があげられ、好ましくは、前記バイオマーカー遺伝子の発現をより強く抑制できることから、４－８－スフィンガジエニンであり、さらに好ましくは、トランス－４－シス－８－スフィンガジエニンである。
 In the present disclosure, the term "sphingoid base" refers to a long-chain alcohol having an amino group, i.e., a long-chain amino alcohol. The sphingoid base may be derived from, for example, an animal or a plant. The sphingoid base may be represented, for example, by the following formula (1). In the following formula (1), R 1 is a saturated or unsaturated hydrocarbon group. Examples of the sphingoid base include sphingosine, sphinganine, sphingadienine, sphingenine, and the like. Examples of the sphingadienine include isomers. Examples of the sphingadienine isomers include 4-8-sphingadienine, in which the double bonds at the 4- and 8-positions are cis:cis, cis:trans, trans:cis, or trans:trans. Examples of the sphingoid base include 4-hydroxy-8-sphingenin. The number of carbon atoms in the sphingoid base is, for example, 12 to 20, and more preferably 18. Specific examples of the sphingoid base include animal-derived sphingoid bases such as C18-sphingosine (d18:1), Sph3), sphinganine (d18:0), and phytosphingosine (t18:0); trans-4-cis-8-sphingadienine (d18:2), Sph1, 4-hydroxy-cis-8-sphingenine (4-Hyd Examples of sphingoid bases derived from plants include C20-sphingosine (C20-sphingosine, (d20:1), Sph4), and the like; preferably, 4-8-sphingadienine is used because it can more strongly suppress the expression of the biomarker gene, and more preferably, trans-4-cis-8-sphingadienine is used.
　本開示において、前記スフィンゴイド塩基は、例えば、生体内で酵素等により分解されることによって、スフィンゴイド塩基を遊離可能な成分であってもよい。前記スフィンゴイド塩基を遊離可能な成分としては、例えば、セラミド、スフィンゴ脂質等があげられる。前記セラミドは、生体内において、セラミダーゼにより分解されることによって、前記スフィンゴイド塩基を遊離する。また、前記スフィンゴ脂質は、生体内において、スフィンゴミエリナーゼまたはグリコシルセラミダーゼにより分解されることによって、セラミドを遊離し、前記セラミドは、さらに、前記セラミダーゼにより分解されることによって、前記スフィンゴイド塩基を遊離する。このため、前記セラミドおよび前記スフィンゴ脂質は、例えば、前記スフィンゴイド塩基の前駆体、前駆物質、またはプロドラッグということもできる。 In the present disclosure, the sphingoid base may be, for example, a component capable of releasing a sphingoid base by being decomposed by an enzyme or the like in a living body. Examples of components capable of releasing a sphingoid base include ceramide and sphingolipids. The ceramide is decomposed by ceramidase in a living body to release the sphingoid base. The sphingolipid is decomposed by sphingomyelinase or glycosylceramidase in a living body to release ceramide, and the ceramide is further decomposed by the ceramidase to release the sphingoid base. For this reason, the ceramide and the sphingolipid can be referred to as, for example, a precursor, precursor substance, or prodrug of the sphingoid base.
　前記「セラミド」は、スフィンゴイド塩基部と、脂肪酸部とから構成され、前記スフィンゴイド塩基部と、前記脂肪酸部とが結合している化合物を意味する。前記セラミドにおいて、前記スフィンゴイド塩基部と前記脂肪酸部とは、下記式（６）で示すように、アミド結合で共有結合している。前記セラミドは、例えば、下記式（６）で表すことができる。前記スフィンゴイド塩基部は、前記セラミドのうち、スフィンゴイド塩基に由来する部分を意味する。前記スフィンゴイド塩基部は、例えば、前記スフィンゴイド塩基の例示において、アミノ基（ＮＨ２）の水素原子が脱離した基があげられる。また、前記脂肪酸部は、前記セラミドうち、脂肪酸に由来する部分を意味する。下記式（６）において、Ｒ２は、例えば、飽和または不飽和の直鎖炭化水素基、末端がω－ヒドロキシ化された飽和または不飽和の直鎖炭化水素基、または末端がエステルω－ヒドロキシ化された飽和または不飽和の直鎖炭化水素基である。前記脂肪酸部における炭素数は、例えば、１６～２４である。前記脂肪酸部の由来は、例えば、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、ネルボン酸（24:1）等があげられる。前記脂肪酸部は、例えば、前記脂肪酸の例示において、ヒドロキシ基（ＯＨ）が脱離した基があげられる。前記セラミドは、例えば、Ｎ－アシルスフィンゴシン（セラミド）、Ｎ－アシルジヒドロスフィンゴシン（ジヒドロセラミド）、Ｎ－アシルフィトスフィンゴシン（フィトセラミド）等があげられる。前記セラミドは、例えば、前記バイオマーカー遺伝子の発現をより強く抑制できることから、好ましくは、前記スフィンゴイド塩基部が、４－８－スフィンガジエニンに由来するセラミドであり、さらに好ましくは、前記スフィンゴイド塩基部が、トランス－４－シス－８－スフィンガジエニンに由来するセラミドである。
 The "ceramide" refers to a compound that is composed of a sphingoid base portion and a fatty acid portion, and in which the sphingoid base portion and the fatty acid portion are bound to each other. In the ceramide, the sphingoid base portion and the fatty acid portion are covalently bound to each other by an amide bond, as shown in the following formula (6). The ceramide can be represented, for example, by the following formula (6). The sphingoid base portion refers to a portion of the ceramide that is derived from a sphingoid base. In the examples of the sphingoid base, the sphingoid base portion can be, for example, a group in which a hydrogen atom of an amino group (NH 2 ) has been eliminated. In addition, the fatty acid portion refers to a portion of the ceramide that is derived from a fatty acid. In the following formula (6), R 2 is, for example, a saturated or unsaturated linear hydrocarbon group, a saturated or unsaturated linear hydrocarbon group having an ω-hydroxylated end, or a saturated or unsaturated linear hydrocarbon group having an ester ω-hydroxylated end. The number of carbon atoms in the fatty acid moiety is, for example, 16 to 24. The fatty acid moiety is derived from, for example, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), and nervonic acid (24:1). The fatty acid moiety is, for example, a group obtained by eliminating a hydroxyl group (OH) from the examples of the fatty acids. The ceramide is, for example, N-acylsphingosine (ceramide), N-acyldihydrosphingosine (dihydroceramide), and N-acylphytosphingosine (phytoceramide). The ceramide is preferably a ceramide in which the sphingoid base moiety is derived from 4-8-sphingadienine, and more preferably a ceramide in which the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, because this can more strongly suppress the expression of the biomarker gene, for example.
　前記スフィンゴ脂質は、前記セラミドが、さらに、糖またはリン酸を含む基と結合した脂質（複合脂質）を意味する。前記スフィンゴ脂質が前記セラミドと糖とが結合した脂質の場合、前記セラミドの第１アルコールと前記糖を含む基とは、グリコシド結合で共有結合している。前記スフィンゴ脂質が前記セラミドとリン酸を含む基とが結合した脂質の場合、前記セラミドの第１アルコールと前記リン酸基を含む基とは、下記式（７）で示すように、エステル結合で共有結合している。前記スフィンゴ脂質は、例えば、スフィンゴ糖脂質およびスフィンゴリン脂質があげられる。前記スフィンゴ脂質は、例えば、下記式（７）で表すことができる。下記式（７）において、Ｒ３が糖に由来する基である場合、前記スフィンゴ脂質は、スフィンゴ糖脂質ということができる。また、下記式（７）において、Ｒ３がリン酸を含む基である場合、前記スフィンゴ脂質は、スフィンゴリン脂質ということができる。
 The sphingolipid means a lipid (complex lipid) in which the ceramide is further bound to a group containing a sugar or phosphoric acid. When the sphingolipid is a lipid in which the ceramide is bound to a sugar, the primary alcohol of the ceramide and the group containing the sugar are covalently bound by a glycoside bond. When the sphingolipid is a lipid in which the ceramide is bound to a group containing a phosphoric acid, the primary alcohol of the ceramide and the group containing the phosphoric acid are covalently bound by an ester bond as shown in the following formula (7). Examples of the sphingolipid include sphingoglycolipids and sphingophospholipids. The sphingolipid can be represented by the following formula (7), for example. In the following formula (7), when R 3 is a group derived from a sugar, the sphingolipid can be called a sphingoglycolipid. In addition, in the following formula (7), when R 3 is a group containing a phosphoric acid, the sphingolipid can be called a sphingophospholipid.
　前記スフィンゴ糖脂質は、例えば、前記セラミドが、さらに、糖部を有する化合物ということもできる。前記糖部は、前記スフィンゴイド糖脂質のうち、糖に由来する部分を意味する。前記糖は、例えば、グルコース、ガラクトース、およびラクトース等があげられる。前記セラミドがグルコースと結合している場合、前記スフィンゴ糖脂質は、グルコシルセラミドという。前記セラミドがガラクトースと結合している場合、前記スフィンゴ糖脂質は、ガラクトシルセラミドという。前記セラミドがラクトースと結合している場合、前記スフィンゴ糖脂質は、ラクトシルセラミドという。前記糖部は、グルコースに由来することが好ましい。 The sphingoglycolipid can be, for example, a compound in which the ceramide further has a sugar moiety. The sugar moiety refers to the portion of the sphingoglycolipid that is derived from a sugar. Examples of the sugar include glucose, galactose, and lactose. When the ceramide is bound to glucose, the sphingoglycolipid is called glucosylceramide. When the ceramide is bound to galactose, the sphingoglycolipid is called galactosylceramide. When the ceramide is bound to lactose, the sphingoglycolipid is called lactosylceramide. The sugar moiety is preferably derived from glucose.
　前記スフィンゴリン脂質は、前記セラミドに、リン酸化合物がエステル結合した化合物ということもできる。前記リン酸化合物は、例えば、ホスホコリン等があげられる。前記セラミドがホスホコリンと結合している場合、前記スフィンゴリン脂質は、スフィンゴミエリンという。 The sphingophospholipid can also be said to be a compound in which a phosphate compound is ester-bonded to the ceramide. An example of the phosphate compound is phosphocholine. When the ceramide is bonded to phosphocholine, the sphingophospholipid is called sphingomyelin.
　前記スフィンゴイド塩基、前記セラミド、または前記スフィンゴ脂質（以下、併せて「スフィンゴイド塩基成分」ともいう）は、単離または精製された化合物でもよいし、スフィンゴイド塩基成分を含む組成物でもよい。また、前記スフィンゴイド塩基成分は、例えば、自家調製してもよいし、市販品を用いてもよい。前記スフィンゴイド塩基として市販品を用いる場合、trans-4-cis-8-sphingadienine（長良サイエンス社）、4-Hydroxy-cis-8-sphingenin（長良サイエンス社）、C18-sphingosine（長良サイエンス社）、C20-sphingosine（長良サイエンス社）等が利用できる。前記セラミドとして市販品を用いる場合、天然植物性グルコシルセラミド（Glucosylceramide、長良サイエンス社（URL：https://www.funakoshi.co.jp/contents/3459））等が利用できる。前記スフィンゴ脂質として市販品を用いる場合、スフィンゴミエリン（オルブラクト　サーダリー　リサーチ　ラボラトリーズ（URL：https://www.funakoshi.co.jp/contents/3311））等が利用できる。前記スフィンゴイド塩基成分を組成物は、例えば、スフィンゴイド塩基成分を含む抽出物；前記抽出物の粗精製物、前記抽出物の乾燥物、前記抽出物の凍結乾燥物、スプレードライ等の前記抽出物の処理物；等があげられる。 The sphingoid base, the ceramide, or the sphingolipid (hereinafter collectively referred to as "sphingoid base component") may be an isolated or purified compound, or a composition containing a sphingoid base component. The sphingoid base component may be, for example, self-prepared or a commercially available product. When a commercially available product is used as the sphingoid base, trans-4-cis-8-sphingadienine (Nagara Science Co., Ltd.), 4-Hydroxy-cis-8-sphingenin (Nagara Science Co., Ltd.), C18-sphingosine (Nagara Science Co., Ltd.), C20-sphingosine (Nagara Science Co., Ltd.), etc. can be used. When a commercially available product is used as the ceramide, natural plant glucosylceramide (Glucosylceramide, Nagara Science Co., Ltd. (URL: https://www.funakoshi.co.jp/contents/3459)) can be used. When using a commercially available product as the sphingolipid, sphingomyelin (Orbract Surdary Research Laboratories (URL: https://www.funakoshi.co.jp/contents/3311)) can be used. Examples of the composition containing the sphingoid base component include an extract containing the sphingoid base component; a crude product of the extract, a dried product of the extract, a freeze-dried product of the extract, a spray-dried product, and the like.
　前記スフィンゴイド塩基成分を含む抽出物は、例えば、前記スフィンゴイド塩基成分を含む植物に対して溶媒抽出を実施することにより製造できる。前記スフィンゴイド塩基を含む植物は、例えば、コメ、トウモロコシ、蒟蒻、大豆、小麦、および酵母等があげられる。前記セラミドを含む植物は、例えば、コメ、トウモロコシ、蒟蒻、大豆、小麦、および酵母等があげられる。前記スフィンゴ脂質を含む植物は、例えば、コメ、トウモロコシ、蒟蒻、大豆、小麦、および酵母等があげられる。前記植物は、１種類を用いてもよいし、２種類以上を用いてもよい。前記抽出に供する植物の材料は、植物個体でもよいし、植物の部分でもよい。前記植物の部分は、例えば、根、根茎、葉、茎、花全草、またはこれらの混合物があげられる。前記材料は、採取した植物そのものでもよいし、乾燥および／または粉砕等を行なった加工品でもよい。前記スフィンゴイド塩基を含む抽出物が、前記セラミドまたは前記スフィンゴ脂質を含む場合、前記スフィンゴイド塩基を含む抽出物は、例えば、さらに、前記抽出物をセラミダーゼと接触させることにより、前記抽出物中の前記セラミドまたは前記スフィンゴ脂質から、前記スフィンゴイド塩基を生成してもよい。 The extract containing the sphingoid base component can be produced, for example, by performing a solvent extraction on a plant containing the sphingoid base component. Examples of the plant containing the sphingoid base include rice, corn, konnyaku, soybeans, wheat, and yeast. Examples of the plant containing the ceramide include rice, corn, konnyaku, soybeans, wheat, and yeast. Examples of the plant containing the sphingolipid include rice, corn, konnyaku, soybeans, wheat, and yeast. One type of plant may be used, or two or more types may be used. The plant material to be subjected to the extraction may be an individual plant or a part of a plant. Examples of the part of the plant include roots, rhizomes, leaves, stems, whole flowers, or a mixture thereof. The material may be the plant itself as collected, or a processed product that has been dried and/or crushed. When the extract containing the sphingoid base contains the ceramide or the sphingolipid, the extract containing the sphingoid base may further be contacted with ceramidase to produce the sphingoid base from the ceramide or the sphingolipid in the extract, for example.
　前記スフィンゴイド塩基成分の抽出に用いる溶媒は、例えば、水、緩衝液等の水性溶媒；メタノール、エタノール、プロピルアルコール、イソプロピルアルコール、ブタノール、イソブタノール等の低級アルコールまたは含水低級アルコール；プロピレングリコール、１，３－ブチレングリコール、１，２－ブチレングリコール、１，４－ブチレングリコール、１，５－ペンタンジオール、１，２－ペンタンジオール、１，３－ペンタンジオール、１，４－ペンタンジオール、１，３，５－ペンタントリオール、グリセリン、ポリエチレングリコール（例えば、分子量１００～１０万）等の多価アルコールまたは含水多価アルコール；アセトン、酢酸エチル、ジエチルエーテル、ジメチルエーテル、エチルメチルエーテル、ジオキサン、ヘキサン、アセトニトリル、キシレン、ベンゼン、クロロホルム、四塩化炭素、フェノール、トルエン等の有機溶媒；適宜規定度を調製した酸（塩酸、硫酸、硝酸、リン酸、ギ酸、酢酸等）またはアルカリ（水酸化ナトリウム、水酸化カリウム、水酸化カルシウム、アンモニア等）；等があげられる。前記溶媒は、１種類を単独で用いてもよいし、２種類以上を併用してもよい。 The solvent used to extract the sphingoid base components is, for example, an aqueous solvent such as water or a buffer solution; a lower alcohol or a water-containing lower alcohol such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, or isobutanol; propylene glycol, 1,3-butylene glycol, 1,2-butylene glycol, 1,4-butylene glycol, 1,5-pentanediol, 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, 1,3,5-pentanetriol, glycol Examples of the solvent include polyhydric alcohols or hydrous polyhydric alcohols such as serine and polyethylene glycol (e.g., molecular weight 100 to 100,000); organic solvents such as acetone, ethyl acetate, diethyl ether, dimethyl ether, ethyl methyl ether, dioxane, hexane, acetonitrile, xylene, benzene, chloroform, carbon tetrachloride, phenol, and toluene; acids (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, etc.) or alkalis (sodium hydroxide, potassium hydroxide, calcium hydroxide, ammonia, etc.) whose normality has been appropriately adjusted; and the like. The solvents may be used alone or in combination of two or more kinds.
　前記処理物における処理は、例えば、酸（塩酸、硫酸、硝酸、リン酸、有機酸等）またはアルカリ（水酸化ナトリウム、水酸化カルシウム、アンモニア等）添加による分解；微生物による発酵または代謝変換；イオン交換樹脂、活性炭、ケイ藻土等による成分吸着；種々の分離モード（イオン交換、親水性吸着、疎水性吸着、サイズ排除、配位子交換、アフィニティー等）を有するクロマトグラフィーを用いた分画；濾紙、メンブランフィルター、または限外濾過膜等を用いた濾過；加圧または減圧；加温または冷却：乾燥または凍結乾燥；ｐＨ調整；脱臭；脱色；長時間の静置保管；等があげられる。前記処理は、１種類を単独で実施でもよいし、２種類以上を実施してもよい。 The treatment of the treated material includes, for example, decomposition by adding acid (hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, organic acid, etc.) or alkali (sodium hydroxide, calcium hydroxide, ammonia, etc.); fermentation or metabolic conversion by microorganisms; component adsorption by ion exchange resins, activated carbon, diatomaceous earth, etc.; fractionation using chromatography with various separation modes (ion exchange, hydrophilic adsorption, hydrophobic adsorption, size exclusion, ligand exchange, affinity, etc.); filtration using filter paper, membrane filters, ultrafiltration membranes, etc.; pressurization or decompression; heating or cooling; drying or freeze-drying; pH adjustment; deodorization; decolorization; long-term static storage; etc. The above treatments may be performed alone or in combination of two or more.
　前記「掻痒」は、痒みを意味する。前記掻痒を伴う皮膚疾患は、例えば、乾燥皮膚、アトピー性皮膚炎、接触皮膚炎、または皮膚真菌感染症等があげられる。 The term "pruritus" refers to itching. Examples of skin diseases accompanied by pruritus include dry skin, atopic dermatitis, contact dermatitis, and fungal skin infections.
　前記「掻痒の抑制」は、痒みが抑制されることを意味する。前記掻痒の抑制は、例えば、直接的または間接的に評価できる。前記直接的な評価の場合、前記掻痒の抑制は、例えば、モデル動物（例えば、アトピー性皮膚炎モデル）またはパッチテスト等を用いて、対象物の投与者における痒みの増悪、悪化、もしくは増強の停止、または痒みの改善により評価できる。前記間接的な評価の場合、前記掻痒の抑制は、例えば、掻痒関連遺伝子の発現の程度を測定することによって評価でき、具体的には、前記掻痒関連遺伝子の発現抑制により評価できる。 The "suppression of itch" means that itch is suppressed. The suppression of itch can be evaluated, for example, directly or indirectly. In the case of the direct evaluation, the suppression of itch can be evaluated, for example, by the cessation of aggravation, worsening, or intensification of itch, or the improvement of itch, in a subject administered with the subject, using a model animal (e.g., an atopic dermatitis model) or a patch test. In the case of the indirect evaluation, the suppression of itch can be evaluated, for example, by measuring the degree of expression of itch-related genes, specifically, by the suppression of expression of the itch-related genes.
　前記掻痒関連遺伝子は、掻痒の惹起、または掻痒が生じやすい皮膚状態の形成に関連する遺伝子を意味する。前記掻痒関連遺伝子は、例えば、ＴＡＲＣ遺伝子、ＴＳＬＰ遺伝子、およびＩＬ－３１ＲＡ遺伝子等があげられる。 The pruritus-related gene refers to a gene that is related to the induction of pruritus or the formation of a skin condition that is prone to pruritus. Examples of the pruritus-related gene include the TARC gene, the TSLP gene, and the IL-31RA gene.
　前記「ＴＡＲＣ（Thymus and Activation-Regulated Chemokine）」は、白血球等の細胞を組織へ遊走させるケモカインの一種である。ＴＡＲＣ遺伝子は、アトピー性皮膚炎等において発現が誘発され、ＴＡＲＣの産生が誘導および増加することが知られている。また、ＴＡＲＣは、アレルギー性炎症を惹起するリンパ球（Ｔｈ２細胞）を誘引し、アレルギー反応を亢進させることが知られている。前記ＴＡＲＣは、アトピー性皮膚炎におけるマーカーとしても利用されている。 The aforementioned "Thymus and Activation-Regulated Chemokine (TARC)" is a type of chemokine that induces migration of cells such as white blood cells to tissues. It is known that the expression of the TARC gene is induced in atopic dermatitis and the like, inducing and increasing the production of TARC. TARC is also known to attract lymphocytes (Th2 cells) that cause allergic inflammation and enhance allergic reactions. The aforementioned TARC is also used as a marker for atopic dermatitis.
　前記ＴＡＲＣ遺伝子の一例として、ヒトＴＡＲＣ遺伝子がコードするｍＲＮＡは、例えば、Genbankにおいてアクセッション番号：ＮＭ＿００２９８７．３で登録されている塩基配列からなるポリヌクレオチド等があげられる。前記ＴＡＲＣ遺伝子の発現量としては、いずれか１または２つ以上のＴＡＲＣ遺伝子のアイソフォームの発現量を測定してもよいし、全てのアイソフォームの発現を測定してもよい。 An example of the TARC gene is the mRNA encoded by the human TARC gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_002987.3. As the expression level of the TARC gene, the expression level of any one or more isoforms of the TARC gene may be measured, or the expression of all isoforms may be measured.
　前記「ＴＳＬＰ（Thymic stromal lymphopoietin、胸腺間質性リンパ球新生因子）」は、サイトカインの一種である。ＴＳＬＰ遺伝子は、アトピー性皮膚炎等において発現が誘発され、ＴＳＬＰの産生が誘導および増加することが知られている。また、ＴＳＬＰは、アレルギー性炎症を惹起するリンパ球（Ｔｈ２細胞）を誘引し、アレルギー反応を亢進させることが知られている。 The aforementioned "TSLP (thymic stromal lymphopoietin)" is a type of cytokine. It is known that the expression of the TSLP gene is induced in atopic dermatitis and the like, inducing and increasing the production of TSLP. TSLP is also known to attract lymphocytes (Th2 cells) that cause allergic inflammation, enhancing allergic reactions.
　前記ＴＳＬＰ遺伝子の一例として、ヒトＴＳＬＰ遺伝子がコードするｍＲＮＡは、例えば、Genbankにおいてアクセッション番号：ＮＭ＿０３３０３５．５で登録されている塩基配列からなるポリヌクレオチド等があげられる。前記ＴＳＬＰ遺伝子の発現量としては、いずれか１または２つ以上のＴＳＬＰ遺伝子のアイソフォームの発現量を測定してもよいし、全てのアイソフォームの発現を測定してもよい。 An example of the TSLP gene is the mRNA encoded by the human TSLP gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_033035.5. As the expression level of the TSLP gene, the expression level of any one or more isoforms of the TSLP gene may be measured, or the expression of all isoforms may be measured.
　前記「ＩＬ－３１ＲＡ（Interleukin-31、インターロイキン３１受容体）」は、Ｔｈ２細胞等から分泌されるＴ細胞サイトカインＩＬ－３１の受容体である。ＩＬ－３１が、ＩＬ－３１ＲＡに結合すると、掻痒を誘発することが知られている。 The aforementioned "IL-31RA (Interleukin-31 receptor)" is a receptor for the T cell cytokine IL-31, which is secreted from Th2 cells and the like. It is known that when IL-31 binds to IL-31RA, it induces pruritus.
　前記ＩＬ－３１ＲＡ遺伝子の一例として、ヒトＩＬ－３１ＲＡ遺伝子がコードするｍＲＮＡは、例えば、Genbankにおいてアクセッション番号：ＮＭ＿００１２４２６３６．２で登録されている塩基配列からなるポリヌクレオチド等があげられる。前記ＩＬ－３１ＲＡ遺伝子の発現量としては、いずれか１または２つ以上のＩＬ－３１ＲＡ遺伝子のアイソフォームの発現量を測定してもよいし、全てのアイソフォームの発現を測定してもよい。 An example of the IL-31RA gene is the mRNA encoded by the human IL-31RA gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_001242636.2. As the expression level of the IL-31RA gene, the expression level of any one or more isoforms of the IL-31RA gene may be measured, or the expression of all isoforms may be measured.
　前記「ＩＬ－８（Interleukin-8、インターロイキン８）」は、好中球を遊走するサイトカイン（ケモカイン）の一種である。ＩＬ－８は、炎症反応の原因となることが知られている。ＩＬ－８は、ＣＸＣＬ８ともいう。 The aforementioned "IL-8 (Interleukin-8)" is a type of cytokine (chemokine) that induces neutrophil migration. IL-8 is known to be the cause of inflammatory reactions. IL-8 is also known as CXCL8.
　前記ＩＬ－８遺伝子の一例として、ヒトＩＬ－８遺伝子がコードするｍＲＮＡは、例えば、Genbankにおいてアクセッション番号：ＮＭ＿００１３５４８４０．３で登録されている塩基配列からなるポリヌクレオチド等があげられる。前記ＩＬ－８遺伝子の発現量としては、いずれか１または２つ以上のＩＬ－８遺伝子のアイソフォームの発現量を測定してもよいし、全てのアイソフォームの発現を測定してもよい。 An example of the IL-8 gene is the mRNA encoded by the human IL-8 gene, which may be, for example, a polynucleotide consisting of the base sequence registered in Genbank under accession number NM_001354840.3. As the expression level of the IL-8 gene, the expression level of any one or more isoforms of the IL-8 gene may be measured, or the expression of all isoforms may be measured.
　本開示において、「遺伝子の発現抑制」は、対象遺伝子の発現量が抑制または低下することを意味し、前記対象遺伝子の発現がある状態から発現がない状態へ変化することを意味してもよい。前記対象遺伝子の発現は、例えば、後述の実施例１または２に準じて、対象遺伝子のｍＲＮＡの発現量を、定量的ＰＣＲにより測定することにより評価できる。前記対象遺伝子が複数のアイソフォームを有する場合、前記対象遺伝子の発現量としては、いずれか１または２つ以上の対象遺伝子のアイソフォームの発現量を測定してもよいし、全てのアイソフォームの発現を測定してもよいが、好ましくは、後者である。 In the present disclosure, "suppression of gene expression" means that the expression level of a target gene is suppressed or reduced, and may mean that the expression of the target gene changes from a state in which it is expressed to a state in which it is not expressed. The expression of the target gene can be evaluated, for example, by measuring the expression level of the mRNA of the target gene by quantitative PCR in accordance with Examples 1 or 2 described below. When the target gene has multiple isoforms, the expression level of the target gene may be measured by measuring the expression level of one or more isoforms of the target gene, or the expression of all isoforms, with the latter being preferred.
　前記「遺伝子」は、ＲＮＡ（例えば、ｍＲＮＡ）の形態、またはＤＮＡの形態（例えば、ｃＤＮＡまたはゲノムＤＮＡ）で存在し得る。ＤＮＡは、二本鎖であっても、一本鎖であってもよい。本明細書において、前記「遺伝子」は、非翻訳領域（ＵＴＲ）の配列等の付加的な配列を含むものであってもよい。 The "gene" may exist in the form of RNA (e.g., mRNA) or DNA (e.g., cDNA or genomic DNA). The DNA may be double-stranded or single-stranded. As used herein, the "gene" may include additional sequences, such as sequences of untranslated regions (UTRs).
　前記「角化細胞（ケラチノサイト）」は、表皮を構成する基底細胞、有棘細胞、顆粒細胞、および角質細胞に分化する能力を有する細胞を意味する。 The term "keratinocytes" refers to cells that have the ability to differentiate into basal cells, spinous cells, granular cells, and keratinocytes that make up the epidermis.
　本開示の掻痒抑制剤によれば、例えば、角化細胞からのロイコトリエンＢ４の産生を抑制できる。前記ロイコトリエンＢ４の産生抑制は、例えば、後述の実施例１（６）に準じて評価できる。本開示の掻痒抑制剤は、例えば、ロイコトリエンＢ４の産生を抑制することにより、掻痒を抑制できる。 The pruritus inhibitor of the present disclosure can, for example, inhibit the production of leukotriene B4 from keratinocytes. The inhibition of leukotriene B4 production can be evaluated, for example, in accordance with Example 1 (6) described below. The pruritus inhibitor of the present disclosure can inhibit pruritus, for example, by inhibiting the production of leukotriene B4.
　本開示の掻痒抑制剤は、例えば、投与対象に対して使用することにより、掻痒を抑制することができる。本開示の掻痒抑制剤の使用条件（投与条件）は、特に制限されず、例えば、投与対象の種類等に応じて、投与形態、投与時期、投与量等を適宜設定できる。 The antipruritic agent of the present disclosure can suppress pruritus, for example, by administering it to a subject. The conditions for use (administration conditions) of the antipruritic agent of the present disclosure are not particularly limited, and the administration form, administration timing, dosage, etc. can be appropriately set depending on, for example, the type of subject to be administered.
　本開示の掻痒抑制剤は、例えば、ｉｎ　ｖｉｖｏで使用してもよいし、ｉｎ　ｖｉｔｒｏで使用してもよい。 The antipruritic agent of the present disclosure may be used, for example, in vivo or in vitro .
　本開示の掻痒抑制剤の投与対象は、特に制限されない。本開示の掻痒抑制剤をｉｎ　ｖｉｖｏで使用する場合、前記投与対象は、例えば、ヒト、またはヒトを除く非ヒト動物があげられる。前記非ヒト動物としては、例えば、マウス、ラット、ウサギ、イヌ、ヒツジ、ウマ、ネコ、ヤギ、サル、モルモット等の哺乳類、鳥類等があげられる。本開示の掻痒抑制剤をｉｎ　ｖｉｔｒｏで使用する場合、前記投与対象は、例えば、細胞、組織、器官等があげられ、前記細胞は、例えば、生体から採取した細胞、培養細胞等があげられ、前記組織または器官は、例えば、生体から採取した組織（生体組織）または器官等があげられる。 The subject of administration of the pruritus suppressor of the present disclosure is not particularly limited. When the pruritus suppressor of the present disclosure is used in vivo , the subject of administration can be, for example, a human or a non-human animal other than a human. The non-human animal can be, for example, a mammal such as a mouse, a rat, a rabbit, a dog, a sheep, a horse, a cat, a goat, a monkey, or a guinea pig, or a bird. When the pruritus suppressor of the present disclosure is used in vitro , the subject of administration can be, for example, a cell, a tissue, or an organ, and the cell can be, for example, a cell collected from a living body, a cultured cell, or the like, and the tissue or organ can be, for example, a tissue (living tissue) or an organ collected from a living body.
　本開示の掻痒抑制剤を含む、下記皮膚外用剤（例えば、経皮投与もしくは皮膚への塗布剤または組成物）または下記経口投与剤もしくは経口投与組成物において、前記スフィンゴイド塩基の配合量は、掻痒抑制効果を奏する範囲、すなわち、有効量であればよい。前記スフィンゴイド塩基の配合量（上限と下限）は、例えば、以下である。なお、下記の例示において、上限値と下限値とは、任意に組合せ可能である。
・下限値：０．１μｇ／ｍｌ
・上限値：５μｇ／ｍｌ、好ましくは１μｇ／ｍｌ In the following skin topical preparations (e.g., transdermal or skin application preparations or compositions) or the following orally administered preparations or compositions containing the antipruritus agent of the present disclosure, the amount of the sphingoid base may be within a range that exhibits an antipruritus effect, i.e., an effective amount. The amount of the sphingoid base (upper and lower limits) is, for example, as follows. In the following examples, the upper and lower limits can be combined in any combination.
Lower limit: 0.1 μg/ml
Upper limit: 5 μg/ml, preferably 1 μg/ml
　本開示の掻痒抑制剤の投与形態は、経口投与または非経口投与があげられる。前記非経口投与は、経皮投与、皮膚への塗布（接触）等があげられる。前記皮膚への塗布は、口腔粘膜への塗布、すなわち、口腔内の上皮細胞への塗布または接触の意味を含んでもよい。また、前記皮膚への塗布は、皮膚表面への塗布に加えてまたは代えて、前記皮膚表面を介した皮膚内または皮下への投与もしくは注入の意味を含んでもよい。前記皮膚表面を介した皮膚内への投与または注入は、例えば、マイクロニードルを用いて実施できる。 The administration form of the antipruritic agent of the present disclosure may be oral or parenteral. Examples of the parenteral administration include transdermal administration and application (contact) to the skin. The application to the skin may include application to the oral mucosa, i.e., application to or contact with epithelial cells in the oral cavity. Furthermore, the application to the skin may include administration or injection into the skin or subcutaneously via the skin surface, in addition to or instead of application to the skin surface. The administration or injection into the skin via the skin surface may be performed, for example, using a microneedle.
　本開示の掻痒抑制剤の剤型は、特に制限されず、例えば、前記投与形態に応じて適宜決定できる。前記剤型は、例えば、液体状、固体状があげられる。前記投与形態が経口投与の場合、前記剤型は、例えば、錠剤、丸剤、カプセル剤、顆粒剤、散剤、液剤等があげられる。 The dosage form of the antipruritic agent of the present disclosure is not particularly limited and can be appropriately determined depending on, for example, the administration form. Examples of the dosage form include liquid and solid forms. When the administration form is oral administration, examples of the dosage form include tablets, pills, capsules, granules, powders, liquids, etc.
　本開示の掻痒抑制剤は、例えば、必要に応じて、添加剤を含んでもよく、組成物として使用する場合、前記添加剤は、薬学的に許容可能な添加剤または薬学的に許容可能な担体を含むことが好ましい。前記添加剤は、特に制限されず、例えば、基剤原料、賦形剤、着色剤、滑沢剤、結合剤、崩壊剤、安定化剤、コーティング剤、保存剤、香料等の矯味矯臭剤等があげられる。本開示において、前記添加剤の配合量は、スフィンゴイド塩基またはセラミド等の機能を妨げるものでなければ、特に制限されない。 The antipruritic agent of the present disclosure may contain, for example, additives as necessary. When used as a composition, the additives preferably contain pharma- ceutically acceptable additives or pharma-ceutically acceptable carriers. The additives are not particularly limited, and examples include base raw materials, excipients, colorants, lubricants, binders, disintegrants, stabilizers, coating agents, preservatives, flavoring agents such as fragrances, etc. In the present disclosure, the amount of the additives to be added is not particularly limited as long as they do not interfere with the function of sphingoid bases or ceramides, etc.
　前記賦形剤は、例えば、乳糖、乳糖水和物、白糖、ブドウ糖、マンニトール、ソルビトール等の糖誘導体；トウモロコシデンプン、バレイショデンプン、αデンプン、デキストリン等のデンプン誘導体；結晶セルロース等のセルロース誘導体；アラビアゴム；デキストラン；プルラン等の有機系賦形剤；軽質無水珪酸、合成珪酸アルミニウム、珪酸カルシウム、メタ珪酸アルミン酸マグネシウム等のケイ酸塩誘導体；リン酸水素カルシウム等のリン酸塩；炭酸カルシウム等の炭酸塩；硫酸カルシウム等の硫酸塩等の無機系賦形剤があげられる。前記着色剤は、例えば、黄色三二酸化鉄等があげられる。前記滑沢剤は、例えば、ステアリン酸、ステアリン酸カルシウム、ステアリン酸マグネシウム等のステアリン酸金属塩；タルク；ポリエチレングリコール；シリカ；硬化植物油等があげられる。前記矯味矯臭剤は、例えば、ココア末、ハッカ脳、芳香散、ハッカ油、竜脳、桂皮末等の香料、甘味料、酸味料等があげられる。前記結合剤は、例えば、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロース、ポリビニルピロリドン、マクロゴール等があげられる。前記崩壊剤は、例えば、カルボキシメチルセルロース、カルボキシメチルセルロースカルシウム等のセルロース誘導体；カルボキシメチルスターチ、カルボキシメチルスターチナトリウム、架橋ポリビニルピロリドン、デンプングリコール酸ナトリウム等の化学修飾デンプンおよび化学修飾セルロース類等があげられる。前記安定化剤は、例えば、メチルパラベン、プロピルパラベン等のパラオキシ安息香酸エステル類；クロロブタノール、ベンジルアルコール、フェニルエチルアルコール等のアルコール類；塩化ベンザルコニウム；フェノール、クレゾール等のフェノール類；チメロサール；デヒドロ酢酸；ソルビン酸等があげられる。前記コーティング剤は、例えば、ヒプロメロース、マクロゴール6000等のマクロゴール、タルク、酸化チタン等があげられる。 Examples of the excipients include sugar derivatives such as lactose, lactose hydrate, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, and dextrin; cellulose derivatives such as crystalline cellulose; organic excipients such as gum arabic, dextran, and pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, and magnesium aluminometasilicate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; and sulfates such as calcium sulfate. Examples of the colorants include yellow ferric oxide. Examples of the lubricants include metal stearates such as stearic acid, calcium stearate, and magnesium stearate; talc; polyethylene glycol; silica; and hydrogenated vegetable oil. Examples of the flavoring agents include flavorings such as cocoa powder, peppermint, aromatic powder, peppermint oil, borneol, and cinnamon powder, as well as sweeteners and acidulants. Examples of the binder include hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinylpyrrolidone, macrogol, etc. Examples of the disintegrant include cellulose derivatives such as carboxymethyl cellulose and calcium carboxymethyl cellulose; chemically modified starches and chemically modified celluloses such as carboxymethyl starch, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, and sodium starch glycolate. Examples of the stabilizer include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; thimerosal; dehydroacetic acid; sorbic acid, etc. Examples of the coating agent include macrogols such as hypromellose and macrogol 6000, talc, titanium oxide, etc.
　本開示の掻痒抑制組成物が経口投与組成物である場合、前記経口投与組成物の具体例は、例えば、飲料、食品、医薬品（類）、医薬部外品（類）等があげられる。 When the anti-pruritus composition of the present disclosure is an orally administered composition, specific examples of the orally administered composition include beverages, foods, medicines (or similar), quasi-drugs (or similar), etc.
　本開示の掻痒抑制剤または組成物が経皮投与または皮膚への塗布に用いられるもの（以下、「皮膚外用剤」ともいう）である場合、前記皮膚外用剤の形態は、利用形態に応じて、アンプル、カプセル、粉末、顆粒、液体、ゲル、気泡、エマルジョン、シート、ミスト、スプレー剤等があげられる。前記利用形態は、例えば、医薬品（類）；医薬部外品（類）；局所用または全身用の皮膚外用剤類；頭皮・頭髪に適用する薬用および／または化粧用の製剤類；浴湯に投じて使用する浴用剤；その他製剤；等があげられる。前記局所用または全身用の皮膚外用剤類は、例えば、化粧水、乳液、クリーム、軟膏、ローション、オイル、パック等の基礎化粧料、固形石鹸、液体ソープ、ハンドウォッシュ等の洗顔料または皮膚洗浄料、マッサージ用剤、クレンジング用剤、除毛剤、脱毛剤、髭剃り処理料、アフターシェーブローション、プレショーブローション、シェービングクリーム、ファンデーション、口紅、頬紅、アイシャドウ、アイライナー、マスカラ等のメークアップ化粧料、香水類、美爪剤、美爪エナメル、美爪エナメル除去剤、パップ剤、プラスター剤、テープ剤、シート剤、貼付剤、エアゾール剤、歯磨きおよびマウスウォッシュ等の含嗽剤（がんそうざい）等があげられる。前記頭皮・頭髪に適用する薬用および／または化粧用の製剤類は、例えば、シャンプー剤、リンス剤、ヘアートリートメント剤、プレヘアートリートメント剤、パーマネント液、染毛料、整髪料、ヘアートニック剤、育毛・養毛料、パップ剤、プラスター剤、テープ剤、シート剤、エアゾール剤等があげられる。前記その他製剤は、例えば、腋臭防止剤または消臭剤、制汗剤、衛生用品、衛生綿類、ウエットティシュ等があげられる。 When the pruritus suppressant or composition disclosed herein is used for transdermal administration or application to the skin (hereinafter also referred to as "topical skin preparation"), the form of the topical skin preparation may be an ampoule, capsule, powder, granule, liquid, gel, foam, emulsion, sheet, mist, spray, etc., depending on the form of use. Examples of the form of use include pharmaceutical product(s); quasi-drug product(s); topical skin preparations for local or whole body use; medicinal and/or cosmetic preparations applied to the scalp and hair; bath additives for use in bath water; other preparations; etc. Examples of the topical or systemic skin preparations include basic cosmetics such as lotions, milky lotions, creams, ointments, lotions, oils and packs; face washes or skin cleansers such as solid soap, liquid soap and hand wash; massage preparations, cleansing preparations, hair removal preparations, depilatories, shaving treatments, aftershave lotions, pre-shave lotions, shaving creams, makeup cosmetics such as foundations, lipsticks, blushers, eye shadows, eyeliners and mascaras; perfumes, nail beautifying preparations, nail beautifying enamel, nail beautifying enamel removers, poultices, plasters, tapes, sheets, patches, aerosols, toothpastes and mouthwashes. Examples of medicinal and/or cosmetic preparations to be applied to the scalp and hair include shampoos, rinses, hair treatments, pre-hair treatments, permanent solutions, hair dyes, hair styling products, hair tonics, hair growth and care products, poultices, plasters, tapes, sheets, aerosols, etc. Examples of other preparations include underarm odor prevention or deodorants, antiperspirants, sanitary products, sanitary cotton, wet tissues, etc.
　前記皮膚外用剤は、必要に応じて、掻痒抑制効果を妨げない範囲で以下に例示する成分および／または添加剤を、任意に選択および／または併用して製造することができる。 The topical skin preparation can be produced by arbitrarily selecting and/or combining the following components and/or additives as necessary, provided that the antipruritic effect is not impaired.
（１）各種油脂類
　アボカド油、アーモンド油、ウイキョウ油、エゴマ油、オリーブ油、オレンジ油、オレンジラファー油、ゴマ油、カカオ脂、カミツレ油、カロット油、キューカンバー油、牛脂脂肪酸、ククイナッツ油、サフラワー油、シア脂、液状シア脂、大豆油、ツバキ油、トウモロコシ油、ナタネ油、パーシック油、ヒマシ油、綿実油、落花生油、タートル油、ミンク油、卵黄油、パーム油、パーム核油、モクロウ、ヤシ油、牛脂、豚脂、スクワレン、スクワラン、プリスタンまたはこれら油脂類の水素添加物（硬化油等）等。 (1) Various Oils and Fats Avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange roughage oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, kukui nut oil, safflower oil, shea butter, liquid shea butter, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, Japan wax, coconut oil, beef tallow, lard, squalene, squalane, pristane, and hydrogenated products of these oils and fats (hardened oils, etc.).
（２）ロウ類
　ミツロウ、カルナバロウ、鯨ロウ、ラノリン、液状ラノリン、還元ラノリン、硬質ラノリン、カンデリラロウ、モンタンロウ、セラックロウ、ライスワックス等。 (2) Waxes: beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax, etc.
（３）鉱物油
　流動パラフィン、ワセリン、パラフィン、オゾケライド、セレシン、マイクロクリスタンワックス等。 (3) Mineral oils: liquid paraffin, vaseline, paraffin, ozokerite, ceresin, microcrystalline wax, etc.
（４）脂肪酸類
　ラウリン酸、ミリスチン酸、パルミチン酸、ステアリン酸、ベヘン酸、オレイン酸、リノール酸、リノレン酸、ドコサヘキサエン酸、エイコサペンタエン酸、１２－ヒドロキシステアリン酸、ウンデシレン酸、トール油、ラノリン脂肪酸等の天然脂肪酸、イソノナン酸、カプロン酸、２－エチルブタン酸、イソペンタン酸、２－メチルペンタン酸、２－エチルヘキサン酸、イソペンタン酸等の合成脂肪酸。 (4) Fatty Acids Natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acid; and synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid, and isopentanoic acid.
（５）アルコール類
　エタノール、イソプロパノール、ラウリルアルコール、セタノール、ステアリルアルコール、オレイルアルコール、ラノリンアルコール、コレステロール、フィトステロール、フェノキシエタノール等の天然アルコール、２－ヘキシルデカノール、イソステアリルアルコール、２－オクチルドデカノール等の合成アルコール。 (5) Alcohols Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, and phenoxyethanol; and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol.
（６）多価アルコール類
　酸化エチレン、エチレングリコール、ジエチレングリコール、トリエチレングリコール、エチレングリコールモノエチルエーテル、エチレングリコールモノブチルエーテル、ジエチレングリコールモノメチルエーテル、ジエチレングリコールモノエチルエーテル、ポリエチレングリコール、酸化プロピレン、プロピレングリコール、ポリプロピレングリコール、１，３－ブチレングリコール、ペンチルグリコール、グリセリン、ペンタエリトリトール、トレイトール、アラビトール、キシリトール、リビトール、ガラクチトール、ソルビトール、マンニトール、ラクチトール、マルチトール等。 (6) Polyhydric Alcohols Ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol, and the like.
（７）エステル類
　ミリスチン酸イソプロピル、パルミチン酸イソプロピル、ステアリン酸ブチル、ラウリン酸ヘキシル、ミリスチン酸ミリスチル、オレイン酸オレイル、オレイン酸デシル、ミリスチン酸オクチルドデシル、ジメチルオクタン酸ヘキシルデシル、乳酸セチル、乳酸ミリスチル、フタル酸ジエチル、フタル酸ジブチル、酢酸ラノリン、モノステアリン酸エチレングリコール、モノステアリン酸プロピレングリコール、ジオレイン酸プロピレングリコール等。 (7) Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, and the like.
（８）金属セッケン類
　ステアリン酸アルミニウム、ステアリン酸マグネシウム、ステアリン酸亜鉛、ステアリン酸カルシウム、パルミチン酸亜鉛、ミリスチン酸マグネシウム、ラウリン酸亜鉛、ウンデシレン酸亜鉛等。 (8) Metallic Soaps Aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, zinc undecylenate, and the like.
（９）ガム質、糖類または水溶性高分子化合物
　アラビアゴム、ベンゾインゴム、ダンマルゴム、グアヤク脂、アイルランド苔、カラヤゴム、トラガントゴム、キャロブゴム、クインシード、寒天、カゼイン、乳糖、果糖、ショ糖またはそのエステル、トレハロースまたはその誘導体、デキストリン、ゼラチン、ペクチン、デンプン、カラギーナン、カルボキシメチルキチンまたはキトサン、エチレンオキサイド等のアルキレン（Ｃ２～Ｃ４）オキサイドが付加されたヒドロキシアルキル（Ｃ２～Ｃ４）キチンまたはキトサン、低分子キチンまたはキトサン、キトサン塩、硫酸化キチンまたはキトサン、リン酸化キチンまたはキトサン、アルギン酸またはその塩、ヒアルロン酸またはその塩、コンドロイチン硫酸またはその塩、ヘパリン、エチルセルロース、メチルセルロース、カルボキシメチルセルロース、カルボキシエチルセルロース、カルボキシエチルセルロースナトリウム、ヒドロキシエチルセルロース、ヒドロキシプロピルセルロース、ニトロセルロース、結晶セルロース、ポリビニルアルコール、ポリビニルメチルエーテル、ポリビニルピロリドン、ポリビニルメタアクリレート、ポリアクリル酸塩、ポリエチレンオキサイドやポリプロピレンオキサイド等のポリアルキレンオキサイドまたはその架橋重合物、カルボキシビニルポリマー、ポリエチレンイミン等。 (9) Gums, sugars or water-soluble polymer compounds Gum arabic, gum benzoin, gum dammar, guaiac butter, Irish moss, gum karaya, gum tragacanth, carob gum, quince seed, agar, casein, lactose, fructose, sucrose or esters thereof, trehalose or derivatives thereof, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C2-C4) chitin or chitosan having an alkylene (C2-C4) oxide such as ethylene oxide added thereto, low molecular weight chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or a salt thereof, Hyaluronic acid or a salt thereof, chondroitin sulfate or a salt thereof, heparin, ethyl cellulose, methyl cellulose, carboxymethyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinylpyrrolidone, polyvinyl methacrylate, polyacrylates, polyalkylene oxides such as polyethylene oxide and polypropylene oxide or crosslinked polymers thereof, carboxyvinyl polymers, polyethyleneimine, and the like.
（１０）界面活性剤
　アニオン界面活性剤（アルキルカルボン酸塩、アルキルスルホン酸塩、アルキル硫酸エステル塩、アルキルリン酸エステル塩）、カチオン界面活性剤（アルキルアミン塩、アルキル四級アンモニウム塩）、両性界面活性剤：カルボン酸型両性界面活性剤（アミノ型、ベタイン型）、硫酸エステル型両性界面活性剤、スルホン酸型両性界面活性剤、リン酸エステル型両性界面活性剤、非イオン界面活性剤（エーテル型非イオン界面活性剤、エーテルエステル型非イオン界面活性剤、エステル型非イオン界面活性剤、ブロックポリマー型非イオン界面活性剤、含窒素型非イオン界面活性剤）、その他の界面活性剤（天然界面活性剤、タンパク質加水分解物の誘導体、高分子界面活性剤、チタン・ケイ素を含む界面活性剤、フッ化炭素系界面活性剤）等。 (10) Surfactants Anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkylamine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylate type amphoteric surfactants (amino type, betaine type), sulfate type amphoteric surfactants, sulfonic acid type amphoteric surfactants, phosphate type amphoteric surfactants, nonionic surfactants (ether type nonionic surfactants, ether ester type nonionic surfactants, ester type nonionic surfactants, block polymer type nonionic surfactants, nitrogen-containing type nonionic surfactants), other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, surfactants containing titanium/silicon, fluorocarbon surfactants), etc.
（１１）各種ビタミン類
　ビタミンＡ群：レチノール、レチナール（ビタミンＡ１）、デヒドロレチナール（ビタミンＡ２）、カロチン、リコピン（プロビタミンＡ）、ビタミンＢ群：チアミン塩酸塩、チアミン硫酸塩（ビタミンＢ１）、リボフラビン（ビタミンＢ２）、ピリドキシン（ビタミンＢ６）、シアノコバラミン（ビタミンＢ１２）、葉酸類、ニコチン酸類、パントテン酸類、ビオチン類、コリン、イノシトール類、ビタミンＣ群：ビタミンＣ酸またはその誘導体、ビタミンＤ群：エルゴカルシフェロール（ビタミンＤ２）、コレカルシフェロール（ビタミンＤ３）、ジヒドロタキステロール、ビタミンＥ群：ビタミンＥまたはその誘導体、ユビキノン類、ビタミンＫ群：フィトナジオン（ビタミンＫ１）、メナキノン（ビタミンＫ２）、メナジオン（ビタミンＫ３）、メナジオール（ビタミンＫ４）、その他、必須脂肪酸（ビタミンＦ）、カルニチン、フェルラ酸、γ－オリザノール、オロット酸、ビタミンＰ類（ルチン、エリオシトリン、ヘスペリジン）、ビタミンＵ等。 (11) Various Vitamins Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), Vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, inositol, Vitamin C group: vitamin C acid or its derivatives, Vitamin D group: ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotachysterol, vitamin E group: vitamin E or its derivatives, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadiol (vitamin K4), essential fatty acids (vitamin F), carnitine, ferulic acid, γ-oryzanol, orotic acid, vitamin P group (rutin, eriocitrin, hesperidin), vitamin U, etc.
（１２）各種アミノ酸類
　バリン、ロイシン、イソロイシン、トレオニン、メチオニン、フェニルアラニン、トリプトファン、リジン、グリシン、アラニン、アスパラギン、グルタミン、セリン、システイン、シスチン、チロシン、プロリン、ヒドロキシプロリン、アスパラギン酸、グルタミン酸、ヒドロキシリジン、アルギニン、オルニチン、ヒスチジン等や、それらの硫酸塩、リン酸塩、硝酸塩、クエン酸塩、或いはピロリドンカルボン酸のごときアミノ酸誘導体等。 (12) Various Amino Acids: valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine, and the like, as well as amino acid derivatives thereof, such as sulfates, phosphates, nitrates, citrates, or pyrrolidone carboxylic acid.
（１３）添加物
　前記皮膚外用剤は、さらに、動物または植物由来の各種添加物を添加してもよい。前記添加物は、例えば、添加しようとする製品種別、形態に応じて常法的に行われる加工を行い、各種の素材から任意に選択して添加できる。前記加工は、例えば、粉砕、製粉、洗浄、加水分解、醗酵、精製、圧搾、抽出、分画、ろ過、乾燥、粉末化、造粒、溶解、滅菌、ｐＨ調整、脱臭、脱色等を任意に選択および／または組み合わせた処理とできる。 (13) Additives The skin topical preparation may further contain various additives derived from animals or plants. The additives may be added by, for example, performing processing that is conventionally performed according to the type and form of the product to which they are to be added, and may be selected from various materials. The processing may be, for example, any of the following processes selected and/or combined: crushing, milling, washing, hydrolysis, fermentation, purification, squeezing, extraction, fractionation, filtration, drying, powdering, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc.
　前記抽出に用いる溶媒は、供する製品の使用目的、種類、または後に行う加工処理等を考慮した上で選択できる。前記抽出溶媒は、例えば、水、メタノール、エタノール、プロピルアルコール、イソプロピルアルコール、ブタノール、イソブタノール等の低級アルコールもしくは含水低級アルコール、プロピレングリコール、１，３－ブチレングリコール、グリセリン等の多価アルコールもしくは含水多価アルコール、アセトン、酢酸エチル等の各種有機溶媒の中から選ばれる１種もしくは２種以上の混液を用いるのが望ましい。ただし、前記抽出溶媒は、例えば、用途により有機溶媒の含有が好ましくない場合においては、水のみを使用したり、もしくは抽出後に除去しやすいエタノールを採用し、単独または水との任意の混液で用いてもよく、または、搾取抽出したものを用いてもよい。 The solvent used for the extraction can be selected taking into consideration the purpose and type of the product to be used, or the subsequent processing. The extraction solvent is preferably one or a mixture of two or more selected from water, lower alcohols or water-containing lower alcohols such as water, methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, and isobutanol, polyhydric alcohols or water-containing polyhydric alcohols such as propylene glycol, 1,3-butylene glycol, and glycerin, and various organic solvents such as acetone and ethyl acetate. However, when the inclusion of an organic solvent is not desirable depending on the application, the extraction solvent may be water alone or ethanol, which is easy to remove after extraction, may be used alone or in any mixture with water, or may be extracted by squeezing.
　前記植物または動物系原料由来の添加物を、全身用または局所用の外用剤、化粧品類に供する場合、前記皮膚外用剤は、例えば、皮膚や頭髪の保護をはじめ、保湿、感触・風合いの改善、柔軟性の付与、刺激の緩和、芳香によるストレスの緩和、細胞賦活（細胞老化防止）、炎症の抑制、肌質・髪質の改善、肌荒れ防止およびその改善、発毛、育毛、脱毛防止、光沢の付与、清浄効果、疲労の緩和、血流促進、温浴効果等の美容的効果のほか、香付け、消臭、増粘、防腐、緩衝等の効果も期待できる。 When the additives derived from plant or animal raw materials are used in external preparations or cosmetics for systemic or local use, the external preparations for the skin can be expected to have, for example, cosmetic effects such as protection of the skin and hair, moisturizing, improving feel and texture, imparting softness, reducing irritation, relieving stress through fragrance, activating cells (preventing cell aging), suppressing inflammation, improving skin and hair quality, preventing and improving rough skin, promoting hair growth, hair care, preventing hair loss, imparting shine, cleansing effects, relieving fatigue, promoting blood flow, and providing a warm bath effect, as well as fragrance, deodorizing, thickening, preserving, and buffering effects.
　前記皮膚外用剤は、例えば、さらにこの他にも、これまでに知られている各原料素材の様々な美容的、薬剤的効果を期待し、これらを組み合わせることによって、本開示の目的とする効果の増進を図り、多機能的な効果を期待した製品とすることも可能である。 The topical skin preparation may, for example, be a product that is expected to have various cosmetic and medicinal effects from the various raw materials that are known to date, and by combining these, it is possible to enhance the intended effect of this disclosure and create a product that is expected to have multifunctional effects.
＜ＴＡＲＣ遺伝子の発現抑制に用いるための剤または組成物＞
　別の態様において、本開示は、ＴＡＲＣ遺伝子を発現抑制可能な剤または組成物を提供する。本開示のＴＡＲＣ遺伝子の発現抑制に用いるための剤は、スフィンゴイド塩基を含む。本開示のＴＡＲＣ遺伝子の発現抑制に用いるための組成物は、スフィンゴイド塩基を含む。本開示のＴＡＲＣ遺伝子の発現抑制剤または組成物によれば、掻痒抑制効果を得ることができる。 <Agent or composition for use in suppressing expression of TARC gene>
In another aspect, the present disclosure provides an agent or composition that can suppress the expression of TARC gene.The agent for use in suppressing the expression of TARC gene of the present disclosure comprises a sphingoid base.The composition for use in suppressing the expression of TARC gene of the present disclosure comprises a sphingoid base.The agent or composition for suppressing the expression of TARC gene of the present disclosure can obtain an antipruritic effect.
　前記「ＴＡＲＣ遺伝子の発現抑制」は、前記ＴＡＲＣ遺伝子の発現量が抑制または低下することを意味し、前記ＴＡＲＣ遺伝子の発現がある状態から発現がない状態へ変化することを意味してもよい。前記ＴＡＲＣ遺伝子の発現は、例えば、後述の実施例１に準じて、ＴＡＲＣ遺伝子のｍＲＮＡの発現量を測定することにより評価できる。 The "inhibition of expression of the TARC gene" means that the expression level of the TARC gene is inhibited or decreased, and may mean that the expression of the TARC gene changes from a state in which it is expressed to a state in which it is not expressed. The expression of the TARC gene can be evaluated, for example, by measuring the expression level of the mRNA of the TARC gene according to Example 1 described below.
　本開示のＴＡＲＣ遺伝子の発現抑制剤は、例えば、投与対象に対して使用することにより、ＴＡＲＣ遺伝子の発現を抑制することができる。これにより、本開示のＴＡＲＣ遺伝子の発現抑制剤は、例えば、掻痒抑制効果を得ることができる。本開示のＴＡＲＣ遺伝子の発現抑制剤の使用条件（投与条件）は、前記本開示の掻痒抑制に用いるための剤の使用条件の説明を援用できる。 The TARC gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the TARC gene. This allows the TARC gene expression inhibitor of the present disclosure to obtain, for example, an effect of suppressing pruritus. The conditions for use (administration conditions) of the TARC gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
＜ＴＳＬＰ遺伝子の発現抑制に用いるための剤または組成物＞
　別の態様において、本開示は、ＴＳＬＰ遺伝子を発現抑制可能な剤または組成物を提供する。本開示のＴＳＬＰ遺伝子の発現抑制に用いるための剤は、スフィンゴイド塩基を含む。本開示のＴＳＬＰ遺伝子の発現抑制に用いるための組成物は、スフィンゴイド塩基を含む。本開示のＴＳＬＰ遺伝子の発現抑制剤または組成物によれば、掻痒抑制効果を得ることができる。 <Agent or composition for use in suppressing expression of TSLP gene>
In another aspect, the present disclosure provides an agent or composition capable of suppressing the expression of the TSLP gene. The agent for use in suppressing the expression of the TSLP gene of the present disclosure comprises a sphingoid base. The composition for use in suppressing the expression of the TSLP gene of the present disclosure comprises a sphingoid base. The agent or composition for suppressing the expression of the TSLP gene of the present disclosure can provide an antipruritic effect.
　前記「ＴＳＬＰ遺伝子の発現抑制」は、前記ＴＳＬＰ遺伝子の発現量が抑制または低下することを意味し、前記ＴＳＬＰ遺伝子の発現がある状態から発現がない状態へ変化することを意味してもよい。前記ＴＳＬＰ遺伝子の発現は、例えば、後述の実施例１または２に準じて、ＴＳＬＰ遺伝子のｍＲＮＡの発現量を測定することにより評価できる。 The "inhibition of expression of the TSLP gene" means that the expression level of the TSLP gene is suppressed or reduced, and may mean that the expression of the TSLP gene changes from a state in which the TSLP gene is expressed to a state in which the TSLP gene is not expressed. The expression of the TSLP gene can be evaluated, for example, by measuring the expression level of the mRNA of the TSLP gene in accordance with Example 1 or 2 described below.
　本開示のＴＳＬＰ遺伝子の発現抑制剤は、例えば、投与対象に対して使用することにより、ＴＳＬＰ遺伝子の発現を抑制することができる。これにより、本開示のＴＳＬＰ遺伝子の発現抑制剤は、例えば、掻痒抑制効果を得ることができる。本開示のＴＳＬＰ遺伝子の発現抑制剤の使用条件（投与条件）は、前記本開示の掻痒抑制に用いるための剤の使用条件の説明を援用できる。 The TSLP gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the TSLP gene. This allows the TSLP gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus. The conditions for use (administration conditions) of the TSLP gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
＜ＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための剤または組成物＞
　別の態様において、本開示は、ＩＬ－３１ＲＡ遺伝子を発現抑制可能な剤または組成物を提供する。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための剤は、スフィンゴイド塩基を含む。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための組成物は、スフィンゴイド塩基を含む。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤または組成物によれば、掻痒抑制効果を得ることができる。 <Agent or composition for use in suppressing expression of IL-31RA gene>
In another aspect, the present disclosure provides an agent or composition capable of suppressing the expression of the IL-31RA gene. The agent for use in suppressing the expression of the IL-31RA gene of the present disclosure comprises a sphingoid base. The composition for use in suppressing the expression of the IL-31RA gene of the present disclosure comprises a sphingoid base. The agent or composition for suppressing the expression of the IL-31RA gene of the present disclosure can provide an effect of suppressing pruritus.
　前記「ＩＬ－３１ＲＡ遺伝子の発現抑制」は、前記ＩＬ－３１ＲＡ遺伝子の発現量が抑制または低下することを意味し、前記ＩＬ－３１ＲＡ遺伝子の発現がある状態から発現がない状態へ変化することを意味してもよい。前記ＩＬ－３１ＲＡ遺伝子の発現は、例えば、後述の実施例１または２に準じて、ＩＬ－３１ＲＡ遺伝子のｍＲＮＡの発現量を測定することにより評価できる。 The "suppression of expression of the IL-31RA gene" means that the expression level of the IL-31RA gene is suppressed or decreased, and may mean that the expression of the IL-31RA gene changes from a state in which it is expressed to a state in which it is not expressed. The expression of the IL-31RA gene can be evaluated, for example, by measuring the expression level of the mRNA of the IL-31RA gene according to Example 1 or 2 described below.
　本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤は、例えば、投与対象に対して使用することにより、ＩＬ－３１ＲＡ遺伝子の発現を抑制することができる。これにより、本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤は、例えば、掻痒抑制効果を得ることができる。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤の使用条件（投与条件）は、前記本開示の掻痒抑制に用いるための剤の使用条件の説明を援用できる。 The IL-31RA gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress the expression of the IL-31RA gene. This allows the IL-31RA gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus. The conditions for use (administration conditions) of the IL-31RA gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
＜ＩＬ－８遺伝子の発現抑制に用いるための剤または組成物＞
　別の態様において、本開示は、ＩＬ－８遺伝子を発現抑制可能な剤または組成物を提供する。本開示のＩＬ－８遺伝子の発現抑制に用いるための剤は、スフィンゴイド塩基を含む。本開示のＩＬ－８遺伝子の発現抑制に用いるための組成物は、スフィンゴイド塩基を含む。本開示のＩＬ－８遺伝子の発現抑制剤または組成物によれば、掻痒抑制効果を得ることができる。 <Agent or composition for use in suppressing expression of IL-8 gene>
In another aspect, the present disclosure provides an agent or composition capable of suppressing the expression of the IL-8 gene. The agent for use in suppressing the expression of the IL-8 gene of the present disclosure comprises a sphingoid base. The composition for use in suppressing the expression of the IL-8 gene of the present disclosure comprises a sphingoid base. The agent or composition for suppressing the expression of the IL-8 gene of the present disclosure can provide an anti-pruritus effect.
　前記「ＩＬ－８遺伝子の発現抑制」は、前記ＩＬ－８遺伝子の発現量が抑制または低下することを意味し、前記ＩＬ－８遺伝子の発現がある状態から発現がない状態へ変化することを意味してもよい。前記ＩＬ－８遺伝子の発現は、例えば、後述の実施例２に準じて、ＩＬ－８遺伝子のｍＲＮＡの発現量を測定することにより評価できる。 The "suppression of expression of the IL-8 gene" means that the expression level of the IL-8 gene is suppressed or decreased, and may mean that the expression of the IL-8 gene changes from a state in which it is expressed to a state in which it is not expressed. The expression of the IL-8 gene can be evaluated, for example, by measuring the expression level of the mRNA of the IL-8 gene according to Example 2 described below.
　本開示のＩＬ－８遺伝子の発現抑制剤は、例えば、投与対象に対して使用することにより、ＩＬ－８遺伝子の発現を抑制することができる。これにより、本開示のＩＬ－８遺伝子の発現抑制剤は、例えば、掻痒抑制効果を得ることができる。本開示のＩＬ－８遺伝子の発現抑制剤の使用条件（投与条件）は、前記本開示の掻痒抑制に用いるための剤の使用条件の説明を援用できる。 The IL-8 gene expression inhibitor of the present disclosure can, for example, be used on a subject to suppress IL-8 gene expression. This allows the IL-8 gene expression inhibitor of the present disclosure to have, for example, an effect of suppressing pruritus. The conditions for use (administration conditions) of the IL-8 gene expression inhibitor of the present disclosure can be the same as those described above for the use conditions of the agent for use in suppressing pruritus.
＜掻痒抑制方法＞
　別の態様において、本開示は、掻痒を抑制可能な方法を開示する。本開示の掻痒抑制方法は、前記本開示の掻痒抑制に用いるための剤および／または組成物を使用する。本開示の掻痒抑制方法によれば、例えば、掻痒抑制効果が得られると期待される。 Method for suppressing pruritus
In another aspect, the present disclosure discloses a method capable of suppressing pruritus. The method for suppressing pruritus of the present disclosure uses the agent and/or composition for use in suppressing pruritus of the present disclosure. According to the method for suppressing pruritus of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained.
　本開示の掻痒抑制方法は、対象に、前記本開示の掻痒抑制に用いるための剤および／または組成物を使用する使用工程を含む。前記使用は、例えば、皮膚等への接触でもよいし、投与でもよい。 The method for suppressing pruritus of the present disclosure includes a step of using the agent and/or composition for use in suppressing pruritus of the present disclosure on a subject. The use may be, for example, contact with the skin or the like, or administration.
　本開示の掻痒抑制方法において、前記使用工程は、例えば、ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで行なってもよい。本開示の掻痒抑制方法の対象（投与対象）および投与条件は、例えば、本開示の掻痒抑制に用いるための剤および／または組成物における投与対象および投与条件の説明を援用できる。 In the method for suppressing pruritus of the present disclosure, the using step may be carried out, for example, in vitro or in vivo . For the subject (administration subject) and administration conditions of the method for suppressing pruritus of the present disclosure, the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus of the present disclosure can be cited, for example.
＜ＴＡＲＣ遺伝子の発現抑制方法＞
　別の態様において、本開示は、ＴＡＲＣ遺伝子の発現を抑制可能な方法を開示する。本開示のＴＡＲＣ遺伝子の発現抑制方法は、前記本開示のＴＡＲＣ遺伝子の発現抑制剤および／または組成物を使用する。本開示のＴＡＲＣ遺伝子の発現抑制方法によれば、例えば、ＴＡＲＣ遺伝子の発現を抑制することにより、掻痒抑制効果が得られると期待される。 <Method for inhibiting expression of TARC gene>
In another aspect, the present disclosure discloses a method for suppressing the expression of TARC gene.The method for suppressing the expression of TARC gene of the present disclosure uses the expression inhibitor and/or composition of the TARC gene of the present disclosure.According to the method for suppressing the expression of TARC gene of the present disclosure, for example, it is expected that the effect of suppressing pruritus can be obtained by suppressing the expression of TARC gene.
　本開示のＴＡＲＣ遺伝子の発現抑制方法は、対象に、前記本開示のＴＡＲＣ遺伝子の発現抑制剤および／または組成物を使用する使用工程を含む。前記使用は、例えば、皮膚等への接触でもよいし、投与でもよい。 The method for suppressing expression of the TARC gene disclosed herein includes a step of using the expression inhibitor and/or composition of the TARC gene disclosed herein on a subject. The use may be, for example, contact with the skin or the like, or administration.
　本開示のＴＡＲＣ遺伝子の発現抑制方法において、前記使用工程は、例えば、ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで行なってもよい。本開示のＴＡＲＣ遺伝子の発現抑制方法の対象（投与対象）および投与条件は、例えば、本開示の掻痒抑制に用いるための剤および／または組成物における投与対象および投与条件の説明を援用できる。 In the method for suppressing the expression of the TARC gene of the present disclosure, the using step may be carried out, for example, in vitro or in vivo . For the subject (administration subject) and administration conditions of the method for suppressing the expression of the TARC gene of the present disclosure, the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus of the present disclosure can be cited, for example.
＜ＴＳＬＰ遺伝子の発現抑制方法＞
　別の態様において、本開示は、ＴＳＬＰ遺伝子の発現を抑制可能な方法を開示する。本開示のＴＳＬＰ遺伝子の発現抑制方法は、前記本開示のＴＳＬＰ遺伝子の発現抑制剤および／または組成物を使用する。本開示のＴＳＬＰ遺伝子の発現抑制方法によれば、例えば、ＴＳＬＰ遺伝子の発現を抑制することにより、掻痒抑制効果が得られると期待される。 <Method for inhibiting expression of TSLP gene>
In another aspect, the present disclosure discloses a method capable of suppressing the expression of the TSLP gene. The method for suppressing the expression of the TSLP gene of the present disclosure uses the expression inhibitor and/or composition of the TSLP gene of the present disclosure. According to the method for suppressing the expression of the TSLP gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing the expression of the TSLP gene.
　本開示のＴＳＬＰ遺伝子の発現抑制方法は、対象に、前記本開示のＴＳＬＰ遺伝子の発現抑制剤および／または組成物を使用する使用工程を含む。前記使用は、例えば、皮膚等への接触でもよいし、投与でもよい。 The method of suppressing expression of the TSLP gene disclosed herein includes a step of using the TSLP gene expression inhibitor and/or composition disclosed herein on a subject. The use may be, for example, contact with the skin or the like, or administration.
　本開示のＴＳＬＰ遺伝子の発現抑制方法において、前記使用工程は、例えば、ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで行なってもよい。本開示のＴＳＬＰ遺伝子の発現抑制方法の対象（投与対象）および投与条件は、例えば、本開示の掻痒抑制に用いるための剤および／または組成物における投与対象および投与条件の説明を援用できる。 In the method for suppressing expression of the TSLP gene disclosed herein, the using step may be carried out, for example, in vitro or in vivo . For the subject (administration subject) and administration conditions of the method for suppressing expression of the TSLP gene disclosed herein, the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
＜ＩＬ－３１ＲＡ遺伝子の発現抑制方法＞
　別の態様において、本開示は、ＩＬ－３１ＲＡ遺伝子の発現を抑制可能な方法を開示する。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制方法は、前記本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤および／または組成物を使用する。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制方法によれば、例えば、ＩＬ－３１ＲＡ遺伝子の発現を抑制することにより、掻痒抑制効果が得られると期待される。 <Method for inhibiting expression of IL-31RA gene>
In another aspect, the present disclosure discloses a method capable of suppressing expression of the IL-31RA gene. The method for suppressing expression of the IL-31RA gene of the present disclosure uses the IL-31RA gene expression inhibitor and/or composition of the present disclosure. According to the method for suppressing expression of the IL-31RA gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing expression of the IL-31RA gene.
　本開示のＩＬ－３１ＲＡ遺伝子の発現抑制方法は、対象に、前記本開示のＩＬ－３１ＲＡ遺伝子の発現抑制剤および／または組成物を使用する使用工程を含む。前記使用は、例えば、皮膚等への接触でもよいし、投与でもよい。 The method for suppressing expression of the IL-31RA gene disclosed herein includes a step of using the IL-31RA gene expression inhibitor and/or composition disclosed herein on a subject. The use may be, for example, contact with the skin or the like, or administration.
　本開示のＩＬ－３１ＲＡ遺伝子の発現抑制方法において、前記使用工程は、例えば、ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで行なってもよい。本開示のＩＬ－３１ＲＡ遺伝子の発現抑制方法の対象（投与対象）および投与条件は、例えば、本開示の掻痒抑制に用いるための剤および／または組成物における投与対象および投与条件の説明を援用できる。 In the method for suppressing expression of the IL-31RA gene disclosed herein, the using step may be carried out, for example, in vitro or in vivo . For the subject (administration subject) and administration conditions of the method for suppressing expression of the IL-31RA gene disclosed herein, the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
＜ＩＬ－８遺伝子の発現抑制方法＞
　別の態様において、本開示は、ＩＬ－８遺伝子の発現を抑制可能な方法を開示する。本開示のＩＬ－８遺伝子の発現抑制方法は、前記本開示のＩＬ－８遺伝子の発現抑制剤および／または組成物を使用する。本開示のＩＬ－８遺伝子の発現抑制方法によれば、例えば、ＩＬ－８遺伝子の発現を抑制することにより、掻痒抑制効果が得られると期待される。 <Method for inhibiting expression of IL-8 gene>
In another aspect, the present disclosure discloses a method capable of suppressing expression of the IL-8 gene. The method for suppressing expression of the IL-8 gene of the present disclosure uses the IL-8 gene expression inhibitor and/or composition of the present disclosure. According to the method for suppressing expression of the IL-8 gene of the present disclosure, for example, it is expected that an effect of suppressing pruritus can be obtained by suppressing expression of the IL-8 gene.
　本開示のＩＬ－８遺伝子の発現抑制方法は、対象に、前記本開示のＩＬ－８遺伝子の発現抑制剤および／または組成物を使用する使用工程を含む。前記使用は、例えば、皮膚等への接触でもよいし、投与でもよい。 The method for suppressing expression of the IL-8 gene disclosed herein includes a step of using the IL-8 gene expression inhibitor and/or composition disclosed herein on a subject. The use may be, for example, contact with the skin or the like, or administration.
　本開示のＩＬ－８遺伝子の発現抑制方法において、前記使用工程は、例えば、ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで行なってもよい。本開示のＩＬ－８遺伝子の発現抑制方法の対象（投与対象）および投与条件は、例えば、本開示の掻痒抑制に用いるための剤および／または組成物における投与対象および投与条件の説明を援用できる。 In the method for suppressing expression of the IL-8 gene disclosed herein, the using step may be carried out, for example, in vitro or in vivo . For the subject (administration subject) and administration conditions of the method for suppressing expression of the IL-8 gene disclosed herein, the explanation of the administration subject and administration conditions of the agent and/or composition for use in suppressing pruritus disclosed herein can be cited, for example.
＜使用＞
　本開示は、掻痒抑制に用いるための、掻痒抑制に用いるための剤および／または組成物の使用である。本開示は、ＴＡＲＣ遺伝子の発現の抑制に用いるための、ＴＡＲＣ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＴＳＬＰ遺伝子の発現の抑制に用いるための、ＴＳＬＰ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための、ＩＬ－３１ＲＡ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＩＬ－８遺伝子の発現の抑制に用いるための、ＩＬ－８遺伝子の発現抑制剤および／または組成物の使用である。 <Use>
The present disclosure relates to the use of an agent and/or composition for use in suppressing pruritus. The present disclosure relates to the use of a TARC gene expression inhibitor and/or composition for use in suppressing TARC gene expression. The present disclosure relates to the use of a TSLP gene expression inhibitor and/or composition for use in suppressing TSLP gene expression. The present disclosure relates to the use of an IL-31RA gene expression inhibitor and/or composition for use in suppressing IL-31RA gene expression. The present disclosure relates to the use of an IL-8 gene expression inhibitor and/or composition for use in suppressing IL-8 gene expression.
　本開示は、掻痒抑制に用いるための剤、または掻痒抑制に用いるための組成物を製造するための、掻痒抑制に用いるための剤および／または組成物の使用である。本開示は、ＴＡＲＣ遺伝子の発現抑制に用いるための剤、またはＴＡＲＣ遺伝子の発現抑制に用いるための組成物を製造するための、ＴＡＲＣ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＴＳＬＰ遺伝子の発現抑制に用いるための剤、またはＴＳＬＰ遺伝子の発現抑制に用いるための組成物を製造するための、ＴＳＬＰ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための剤、またはＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための組成物を製造するための、ＩＬ－３１ＲＡ遺伝子の発現抑制剤および／または組成物の使用である。本開示は、ＩＬ－８遺伝子の発現抑制に用いるための剤、またはＩＬ－８遺伝子の発現抑制に用いるための組成物を製造するための、ＩＬ－８遺伝子の発現抑制剤および／または組成物の使用である。 The present disclosure relates to the use of an agent and/or composition for use in suppressing pruritus, for producing an agent for use in suppressing pruritus, or a composition for use in suppressing pruritus. The present disclosure relates to the use of an agent and/or composition for suppressing TARC gene expression, for producing an agent for use in suppressing TARC gene expression, or a composition for use in suppressing TARC gene expression. The present disclosure relates to the use of an agent and/or composition for suppressing TSLP gene expression, for producing an agent for use in suppressing TSLP gene expression, or a composition for use in suppressing TSLP gene expression. The present disclosure relates to the use of an agent and/or composition for suppressing IL-31RA gene expression, for producing an agent for use in suppressing IL-31RA gene expression, or a composition for use in suppressing IL-31RA gene expression. The present disclosure relates to the use of an agent and/or composition for suppressing IL-8 gene expression, for producing an agent for use in suppressing IL-8 gene expression, or a composition for use in suppressing IL-8 gene expression.
　つぎに、本発明の実施例について説明する。ただし、本発明は、以下の実施例により制限されない。市販の試薬は、特に示さない限り、それらのプロトコルに基づいて使用した。なお、「ｍｏｌ／ｌ」は、「Ｍ」と標記することもある。 Next, examples of the present invention will be described. However, the present invention is not limited to the following examples. Commercially available reagents were used according to their protocols unless otherwise specified. "mol/l" may also be abbreviated as "M."
［実施例１］
　スフィンゴイド塩基が、掻痒抑制効果を示すことを確認した。 [Example 1]
It was confirmed that sphingoid bases exhibit an antipruritic effect.
（１）ＴＡＲＣ産生試験
　２４ウェルプレートに、ヒト角化細胞であるＨａＣａＴ細胞を播種した。培地は、１０％ＦＢＳ含有ＤＭＥＭ培地（041-29775、富士フイルム和光純薬社製、実施例において以下同じ）を使用した。前記播種後、前記ＨａＣａＴ細胞が９割コンフルエントになるまで培養した。前記培養後、各ウェルの培地を、無血清のＤＭＥＭに交換した。前記交換後、前記ＨａＣａＴ細胞を２４時間培養した。前記培養後、１μｇ／ｍｌのフィトセラミド（Ｆｃｅｒ、フィトセラ　NW-10、一丸ファルコス社製）、０．１μｇ／ｍｌのグルコシルセラミド（Ｇｃｅｒ、長良サイエンス社製）、または１μｇ／ｍｌのスフィンゴイド塩基類（Ｓｐｈ１：trans-4-cis-8-sphingadienine、Ｓｐｈ２：4-Hydroxy-cis-8-sphingenin、長良サイエンス社製）を添加し、１時間培養した。前記培養後、刺激因子であるＴＮＦαおよびＩＦＮγをそれぞれ１０ｎｇ／ｍｌになるように添加し、１６時間培養した。前記培養後、培養上清を回収し、Human IL-6 Quantikine ELISA Kit（R&D System社製）を用いて、ＥＬＩＳＡを行った。前記ＥＬＩＳＡの結果から、ＴＡＲＣの産生量を定量化した。各群は、３サンプルで実施した（ｎ＝３）。なお、統計解析はＴｕｒｋｙ’ｓ　ｔｅｓｔにより実施した。これらの結果を図１に示す。 (1) TARC Production Test HaCaT cells, which are human keratinocytes, were seeded on a 24-well plate. The medium used was a 10% FBS-containing DMEM medium (041-29775, Fujifilm Wako Pure Chemical Industries, Ltd., the same applies below in the Examples). After the seeding, the HaCaT cells were cultured until they were 90% confluent. After the culture, the medium in each well was replaced with serum-free DMEM. After the replacement, the HaCaT cells were cultured for 24 hours. After the culture, 1 μg/ml of phytoceramide (Fcer, Phytocera NW-10, Ichimaru Falcos), 0.1 μg/ml of glucosylceramide (Gcer, Nagara Science), or 1 μg/ml of sphingoid bases (Sph1: trans-4-cis-8-sphingadienine, Sph2: 4-Hydroxy-cis-8-sphingenin, Nagara Science) were added and cultured for 1 hour. After the culture, stimulating factors TNFα and IFNγ were added to 10 ng/ml each and cultured for 16 hours. After the culture, the culture supernatant was collected and ELISA was performed using Human IL-6 Quantikine ELISA Kit (R&D System). The amount of TARC produced was quantified from the results of the ELISA. Each group was performed with 3 samples (n=3). Statistical analysis was performed using the Turkey's test. The results are shown in FIG.
　図１は、ＨａＣａＴ細胞における、ＴＡＲＣ産生量を示すグラフである。図１において、縦軸は、ＴＡＲＣ産生量（ｐｇ／ｍｌ）を示し、横軸は、試料の種類を示す。
　図１における縦軸の数値は、細胞数で補正したＴＡＲＣ産生量として、各群の値を示している。
　図１の縦軸の数値は、以下の通りである。
・Control群：6.59、No add群：695.05、1μg/ml Fcer群：596.88、0.1μg/ml Gcer群：665.62、1μg/ml Sph1群：590.90、1μg/ml Sph2群：646.62
 
　図１に示すように、Ｆｃｅｒ（コメ由来グルコシルセラミド）群またはＳｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＡＲＣ産生量が有意に減少した。これらの結果から、角化細胞において、コメ由来グルコシルセラミドおよびＳｐｈ１がＴＡＲＣ産生を抑制することがわかった。 Fig. 1 is a graph showing the amount of TARC produced in HaCaT cells, in which the vertical axis indicates the amount of TARC produced (pg/ml) and the horizontal axis indicates the type of sample.
The values on the vertical axis in FIG. 1 indicate the values for each group as the amount of TARC produced corrected by the cell number.
The values on the vertical axis in FIG. 1 are as follows:
Control group: 6.59, No add group: 695.05, 1μg/ml Fcer group: 596.88, 0.1μg/ml Gcer group: 665.62, 1μg/ml Sph1 group: 590.90, 1μg/ml Sph2 group: 646.62

As shown in Figure 1, the amount of TARC produced was significantly reduced in the Fcer (rice-derived glucosylceramide) group or Sph1 group compared to the No add (negative control) group. These results demonstrated that rice-derived glucosylceramide and Sph1 suppress TARC production in keratinocytes.
（２）ＴＳＬＰの遺伝子発現量の試験（Ｉ）
　角化細胞において、コメ由来グルコシルセラミドおよびＳｐｈ１が、ＴＡＲＣ産生抑制効果を示すことがわかったが、通常の角化細胞においてＴＡＲＣ産生は生じない現象であるという報告がある。そこで、掻痒のバイオマーカーの一種として用いられているＴＳＬＰの遺伝子発現の試験を行った。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ヒト角化細胞であるＮＨＥＫ細胞（KK-4009、凍結NHEK(NB)、新生児由来、クラボウ社製）を播種した。培地は、ＫＢＭ（商標）培地（カルシウム・フェノールレッド不含、00195769、ロンザ株式会社製、以下実施例において同じ。）を使用した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍｍｏｌ／ｌカルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、１μｇ／ｍｌの濃度のＦｃｅｒまたはＳｐｈ１を添加し、１時間培養した。前記培養後、刺激因子であるＴＮＦαを１０ｎｇ／ｍｌ（最終濃度）およびＩＮＦγを１０ｎｇ／ｍｌ（最終濃度）となるように添加し、６時間培養した。前記培養後、培養上清を回収し、RNeasy Mini Kit（QIAGEN社製）を用いて、添付のプロトコルに従ってｍＲＮＡを精製した。前記精製後、PrimeScript Master Mix（TaKaRa社製）を用いて逆転写反応を行った。つぎに、TB Green Master Mix（TaKaRa社製）記載の方法に従い、下記ＴＳＬＰ遺伝子用プライマーセットおよびＲＰＳ遺伝子用プライマーセットを用いて、ｑＲＴ－ＰＣＲを行った。ＰＣＲ装置には、LightCycler（登録商標）96（Roche社製）を用いた。ＴＳＬＰ遺伝子の発現量は、内部標準遺伝子（ＲＰＳ１８遺伝子）の発現量に対する相対値とした。データの解析は、付属のソフトウェアを用いて解析した。統計解析は、ＪＭＰ８を用い、Ｔｕｒｋｙ検定およびＤｕｎｎｅｔｔ’ｓ　ｔｅｓｔにより実施した。これらの結果を図２に示す。 (2) Test of TSLP gene expression level (I)
It has been found that rice-derived glucosylceramide and Sph1 have an inhibitory effect on TARC production in keratinocytes, but it has been reported that TARC production does not occur in normal keratinocytes. Therefore, a test was performed on the gene expression of TSLP, which is used as a kind of biomarker for pruritus. Specifically, human keratinocytes, NHEK cells (KK-4009, frozen NHEK (NB), derived from newborns, manufactured by Kurabo Industries, Ltd.) were seeded on a 24-well plate at 1.0 x 10 5 cells/well. The medium used was KBM (trademark) medium (calcium-phenol red-free, 00195769, manufactured by Lonza Co., Ltd., the same applies in the following examples). After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM (trademark) medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotics (GA-1000), and 0.06 mmol/l calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, Fcer or Sph1 was added at a concentration of 1 μg/ml, and the cells were cultured for 1 hour. After the culture, the stimulatory factors TNFα and INFγ were added to 10 ng/ml (final concentration) and 10 ng/ml (final concentration), respectively, and the cells were cultured for 6 hours. After the culture, the culture supernatant was collected, and mRNA was purified using RNeasy Mini Kit (QIAGEN) according to the attached protocol. After the purification, reverse transcription reaction was performed using PrimeScript Master Mix (TaKaRa). Next, qRT-PCR was performed using the following primer set for the TSLP gene and primer set for the RPS gene according to the method described in TB Green Master Mix (manufactured by TaKaRa). The PCR device used was LightCycler (registered trademark) 96 (manufactured by Roche). The expression level of the TSLP gene was taken as a relative value to the expression level of the internal standard gene (RPS18 gene). Data was analyzed using the accompanying software. Statistical analysis was performed using JMP8 by the Turky test and Dunnett's test. These results are shown in FIG. 2.
ＴＳＬＰ遺伝子用プライマーセット
　フォワードプライマー（配列番号１）
　　5’-TAGCAATCGGCCACATTGCCT-3’
　リバースプライマー（配列番号２）
　　5’-GAAGCGACGCCACAATCCTTG-3’
ＲＰＳ１８遺伝子用プライマーセット
　フォワードプライマー（配列番号５）
　　5’-TTTGCGAGTACTCAACACCAACA-3’
　リバースプライマー（配列番号６）
　　5’-GAGCATATCTTCGGCCCACAC-3’ Primer set for TSLP gene Forward primer (SEQ ID NO: 1)
5'-TAGCAATCGGCCACATTGCCT-3'
Reverse primer (SEQ ID NO:2)
5'-GAAGCGACGCCACAATCCTTG-3'
Primer set for RPS18 gene Forward primer (SEQ ID NO: 5)
5'-TTTGCAGTACTCAACACCAACA-3'
Reverse primer (SEQ ID NO:6)
5'-GAGCATATCTTCGGCCCACAC-3'
　図２は、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。図２（Ａ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図２（Ｂ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。
　図２（Ａ）および（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図２（Ａ）の縦軸の数値は、以下の通りである。
・No add群：6.99、1μg/ml Fcer群：7.03
　図２（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：6.99、1μg/ml Sph1群：4.72
 
　図２（Ａ）に示すように、Ｆｃｅｒ群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量は変わらなかった。これに対して、図２（Ｂ）に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１は、ＴＳＬＰの遺伝子発現を抑制することがわかった。 Fig. 2 is a graph showing the expression level of the TSLP gene in NHEK cells. In Fig. 2(A), the vertical axis shows the relative value of the expression level of the TSLP gene, and the horizontal axis shows the type of sample. In Fig. 2(B), the vertical axis shows the relative value of the expression level of the TSLP gene, and the horizontal axis shows the type of sample.
The values on the vertical axis in Figures 2 (A) and (B) indicate the values for each group, with the value for the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 6.99, 1μg/ml Fcer group: 7.03
The values on the vertical axis in FIG.
No add group: 6.99, 1μg/ml Sph1 group: 4.72

As shown in Figure 2(A), the gene expression level of TSLP was unchanged in the Fcer group compared to the No add (negative control) group. In contrast, as shown in Figure 2(B), the gene expression level of TSLP was significantly suppressed in the Sph1 group compared to the No add (negative control) group. From the above, it was found that Sph1 suppresses the gene expression of TSLP in keratinocytes.
（３）ＴＳＬＰの遺伝子発現量の試験（ＩＩ）
　つぎに、炎症性サイトカイン（ＴＮＦαおよびＩＮＦγ）の代わりに、ＴＬＲ３に作用してＴＳＬＰ産生を促すＰｏｌｙ（Ｉ：Ｃ）を用いて、ＴＳＬＰの遺伝子発現の試験を行った。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ＮＨＥＫ細胞を播種した。培地は、ＫＢＭ（商標）培地を使用した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍＭカルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、１μｇ／ｍｌのフィトセラミド（Ｆｃｅｒ）またはスフィンゴイド塩基類（Ｓｐｈ１）を添加し、１時間培養した。前記培養後、Ｐｏｌｙ（Ｉ：Ｃ）を１μｇ／ｍｌになるように添加し、１６時間培養した。前記培養後、培養上清を回収し、RNeasy Mini Kit（QIAGEN社製）を用いて、記載のプロトコルに従ってｍＲＮＡを精製した。前記精製後、PrimeScript Master Mix（TaKaRa社製）を用いて逆転写反応を行った。つぎに、TB Green Master Mix（TaKaRa社製）記載の方法に従い、前記ＴＳＬＰ遺伝子用プライマーセットおよび前記ＲＰＳ遺伝子用プライマーセットを用いて、ｑＲＴ－ＰＣＲを行った。ＰＣＲ装置には、LightCycler（登録商標）96（Roche社製）を用いた。前記ＴＳＬＰ遺伝子の遺伝子は、内部標準遺伝子（ＲＰＳ１８遺伝子）の発現量に対する相対値とした。また、タンパク質レベルでの検出は、前記培養上清回収後、遠心分離を行い、デブリを除去した。前記除去後、Human TSLP Quantikine ELISA Kit（R&D System社製）を用いて、ＥＬＩＳＡを行った。前記ＥＬＩＳＡの結果から、ＴＳＬＰの産生量を定量化した。統計解析は、ＪＭＰ８を用い、Ｔｕｒｋｙ検定により実施した。これらの結果を図３および図４に示す。 (3) Test of TSLP gene expression level (II)
Next, instead of inflammatory cytokines (TNFα and INFγ), Poly(I:C) that acts on TLR3 to promote TSLP production was used to test the gene expression of TSLP. Specifically, NHEK cells were seeded in a 24-well plate at 1.0×10 5 cells/well. KBM (trademark) medium was used. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM (trademark) medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotic (GA-1000), and 0.06 mM calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, 1 μg/ml of phytoceramide (Fcer) or sphingoid bases (Sph1) were added and cultured for 1 hour. After the culture, Poly(I:C) was added to 1 μg/ml, and cultured for 16 hours. After the culture, the culture supernatant was collected, and mRNA was purified using RNeasy Mini Kit (QIAGEN) according to the protocol described. After the purification, reverse transcription reaction was performed using PrimeScript Master Mix (TaKaRa). Next, qRT-PCR was performed using the primer set for the TSLP gene and the primer set for the RPS gene according to the method described in TB Green Master Mix (TaKaRa). LightCycler (registered trademark) 96 (Roche) was used as the PCR device. The gene expression of the TSLP gene was a relative value to the expression level of the internal standard gene (RPS18 gene). In addition, for detection at the protein level, centrifugation was performed after the culture supernatant was collected, and debris was removed. After the removal, ELISA was performed using Human TSLP Quantikine ELISA Kit (R&D System). The amount of TSLP produced was quantified from the results of the ELISA. Statistical analysis was performed using the Turky test with JMP 8. The results are shown in Figures 3 and 4.
　図３は、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。図３（Ａ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図３（Ｂ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。
　図３（Ａ）および（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図３（Ａ）の縦軸の数値は、以下の通りである。
・No add群：389.03、1μg/ml Fcer群：331.34
　図３（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：389.03、1μg/ml Sph1群：231.40
 
　図３（Ａ）に示すように、Ｆｃｅｒ群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量は変わらなかった。これに対して、図３（Ｂ）に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１は、ＴＳＬＰの遺伝子発現を抑制することがわかった。 3 is a graph showing the expression level of TSLP gene in NHEK cells. In Fig. 3(A), the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample. In Fig. 3(B), the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
The values on the vertical axis in Figures 3 (A) and (B) indicate the values for each group, with the value for the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 389.03, 1μg/ml Fcer group: 331.34
The values on the vertical axis in FIG.
No add group: 389.03, 1μg/ml Sph1 group: 231.40

As shown in Figure 3(A), the gene expression level of TSLP was unchanged in the Fcer group compared to the No add (negative control) group. In contrast, as shown in Figure 3(B), the gene expression level of TSLP was significantly suppressed in the Sph1 group compared to the No add (negative control) group. From the above, it was found that Sph1 suppresses the gene expression of TSLP in keratinocytes.
　図４は、ＮＨＥＫ細胞における、ＴＳＬＰ産生量を示すグラフである。図４（Ａ）において、縦軸は、ＴＳＬＰ産生量を示し、横軸は、試料の種類を示す。図４（Ｂ）において、縦軸は、ＴＳＬＰ産生量を示し、横軸は、試料の種類を示す。
　図４における縦軸の数値は、細胞数で補正したＴＳＬＰ産生量として、各群の値を示している。
　図４（Ａ）の縦軸の数値は、以下の通りである。
・Control群：9.02、No add群：56.51、1μg/ml Fcer群：54.69
　図４（Ｂ）の縦軸の数値は、以下の通りである。
・Control群：9.02、No add群：56.51、1μg/ml Sph1群：42.68
 
　図４（Ａ）に示すように、Ｆｃｅｒ群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの産生量は変わらなかった。これに対して、図４（Ｂ）に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの産生量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１は、ＴＳＬＰ産生を抑制することがわかった。 Fig. 4 is a graph showing the amount of TSLP produced in NHEK cells. In Fig. 4(A), the vertical axis shows the amount of TSLP produced, and the horizontal axis shows the type of sample. In Fig. 4(B), the vertical axis shows the amount of TSLP produced, and the horizontal axis shows the type of sample.
The values on the vertical axis in FIG. 4 indicate the values for each group as the amount of TSLP produced corrected by the cell number.
The values on the vertical axis in FIG.
Control group: 9.02, No add group: 56.51, 1μg/ml Fcer group: 54.69
The values on the vertical axis in FIG.
Control group: 9.02, No add group: 56.51, 1μg/ml Sph1 group: 42.68

As shown in Figure 4(A), the amount of TSLP produced in the Fcer group was unchanged compared to the No add (negative control) group. In contrast, as shown in Figure 4(B), the amount of TSLP produced in the Sph1 group was significantly suppressed compared to the No add (negative control) group. From the above, it was found that Sph1 suppresses TSLP production in keratinocytes.
（４）ＩＬ－３１ＲＡの遺伝子発現量の試験
　表皮細胞にはＩＬ－３１受容体Ａ（ＩＬ－３１ＲＡ）が発現しており、表皮細胞へのＩＬ－３１添加はロイコトリエンＢ４（ＬＴＢ４）の産生を促進すること、ＩＬ－３１ＲＡ抗体や阻害剤等が掻痒を改善およびＬＴＢ４産生を抑制することが報告されている。そこで、Ｓｐｈ１が、ＴＳＬＰの遺伝子発現の抑制およびタンパク質産生の抑制に加えて、ＩＬ－３１ＲＡの遺伝子発現の抑制に寄与するか検討した。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ＮＨＥＫ細胞を播種した。培地は、ＫＢＭ（商標）培地を使用した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍｍｏｌ／ｌ　カルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、０．１μｇ／ｍｌ、０．５μｇ／ｍｌ、または１μｇ／ｍｌのフィトセラミド（Ｆｃｅｒ）またはスフィンゴイド塩基類（Ｓｐｈ１）を添加し、１時間培養した。前記培養後、ＴＮＦαおよびＩＮＦγを１０ｎｇ／ｍｌ（最終濃度）になるように添加し、６時間培養した。前記培養後、培養上清を回収し、RNeasy Mini Kit（QIAGEN社製）を用いて、記載のプロトコルに従ってｍＲＮＡを精製した。前記精製後、PrimeScript Master Mix（TaKaRa社製）を用いて逆転写反応を行った。つぎに、TB Green Master Mix（TaKaRa社製）記載の方法に従い、下記ＩＬ－３１ＲＡ遺伝子用プライマーセットおよび前記ＲＰＳ遺伝子用プライマーセットを用いて、ｑＲＴ－ＰＣＲを行った。ＰＣＲ装置には、LightCycler（登録商標）96（Roche社製）を用いた。前記ＩＬ－３１ＲＡ遺伝子の発現量は、内部標準遺伝子（ＲＰＳ１８遺伝子）の発現量に対する相対値とした。データの解析は、付属のソフトウェアを用いた。統計解析は、ＪＭＰ８を用い、Ｔｕｒｋｙ検定により実施した。これらの結果を図５に示す。 (4) Test of gene expression level of IL-31RA It has been reported that IL-31 receptor A (IL-31RA) is expressed in epidermal cells, that the addition of IL-31 to epidermal cells promotes the production of leukotriene B4 (LTB4), and that IL-31RA antibodies and inhibitors improve itching and suppress LTB4 production. Therefore, we investigated whether Sph1 contributes to the suppression of IL-31RA gene expression in addition to the suppression of TSLP gene expression and protein production. Specifically, NHEK cells were seeded in a 24-well plate at 1.0 x 10 5 cells/well. KBM (trademark) medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM (trademark) medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotics (GA-1000), and 0.06 mmol/l calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, 0.1 μg/ml, 0.5 μg/ml, or 1 μg/ml of phytoceramide (Fcer) or sphingoid bases (Sph1) were added and cultured for 1 hour. After the culture, TNFα and INFγ were added to a final concentration of 10 ng/ml and cultured for 6 hours. After the culture, the culture supernatant was collected, and mRNA was purified using RNeasy Mini Kit (QIAGEN) according to the protocol described. After the purification, reverse transcription reaction was performed using PrimeScript Master Mix (TaKaRa). Next, qRT-PCR was performed using the following primer set for the IL-31RA gene and the primer set for the RPS gene according to the method described in TB Green Master Mix (manufactured by TaKaRa). The PCR device used was a LightCycler (registered trademark) 96 (manufactured by Roche). The expression level of the IL-31RA gene was determined as a relative value to the expression level of the internal standard gene (RPS18 gene). Data was analyzed using the accompanying software. Statistical analysis was performed by the Turky test using JMP8. These results are shown in FIG. 5.
ＩＬ－３１ＲＡ遺伝子用プライマーセット
　フォワードプライマー（配列番号３）
　　5’-TGAGTTGGCGCCTGTTTCAT-3’
　リバースプライマー（配列番号４）
　　5’-TTTGACTCCTTGACCGCACA-3’ Primer set for IL-31RA gene Forward primer (SEQ ID NO: 3)
5'-TGAGTTGGCGCCTGTTTCAT-3'
Reverse primer (SEQ ID NO: 4)
5'-TTTGACTCCTTGACCGCACA-3'
　図５は、ＮＨＥＫ細胞における、ＩＬ－３１ＲＡの遺伝子発現量を示すグラフである。図５（Ａ）において、縦軸は、ＩＬ－３１ＲＡ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図５（Ｂ）において、縦軸は、ＩＬ－３１ＲＡ遺伝子の発現量の相対値を示し、横軸は、添加したＳｐｈ１の濃度を示す。
　図５（Ａ）および（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図５（Ａ）の縦軸の数値は、以下の通りである。
・No add群：18.76、0.1μg/ml Fcer群：17.68、0.5μg/ml Fcer群：15.76、1μg/ml Fcer群：15.38
　図５（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：18.76、0.1μg/ml Sph1群：15.05、0.5μg/ml Sph1群：12.55、1μg/ml Sph1群：10.06
 
　図５（Ａ）に示すように、Ｆｃｅｒ群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、Ｆｃｅｒを０．５μｇ／ｍｌまたは１μｇ／ｍｌの濃度で添加した場合、ＩＬ－３１ＲＡの遺伝子発現量が有意に抑制された。また、図５（Ｂ）に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、０．１μｇ／ｍｌ、０．５μｇ／ｍｌ、または１μｇ／ｍｌの濃度で添加した場合、ＩＬ－３１ＲＡの遺伝子発現量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１は、ＩＬ－３１ＲＡの遺伝子発現を抑制することがわかった。 Fig. 5 is a graph showing the expression level of IL-31RA gene in NHEK cells. In Fig. 5(A), the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample. In Fig. 5(B), the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the concentration of Sph1 added.
The numerical values on the vertical axis in Figures 5 (A) and (B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 18.76, 0.1 μg / ml Fcer group: 17.68, 0.5 μg / ml Fcer group: 15.76, 1 μg / ml Fcer group: 15.38
The values on the vertical axis in FIG.
No add group: 18.76, 0.1 μg/ml Sph1 group: 15.05, 0.5 μg/ml Sph1 group: 12.55, 1 μg/ml Sph1 group: 10.06

As shown in Figure 5(A), in the Fcer group, the gene expression level of IL-31RA was significantly suppressed when Fcer was added at a concentration of 0.5 µg/ml or 1 µg/ml, compared to the No add (negative control) group. Also, as shown in Figure 5(B), in the Sph1 group, the gene expression level of IL-31RA was significantly suppressed when Fcer was added at a concentration of 0.1 µg/ml, 0.5 µg/ml, or 1 µg/ml, compared to the No add (negative control) group. From the above, it was found that Sph1 suppresses the gene expression of IL-31RA in keratinocytes.
（５）ロイコトリエンＢ４の産生試験
　Ｓｐｈ１によるＩＬ－３１ＲＡの遺伝子発現の抑制が、掻痒を引き起こすロイコトリエンＢ４（ＬＴＢ４）の産生を抑制するか検討した。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ＮＨＥＫ細胞を播種した。培地は、ＫＢＭ（商標）培地を使用した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍＭカルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、０．５μｇ／ｍｌまたは１μｇ／ｍｌのスフィンゴイド塩基類（Ｓｐｈ１）を添加し、１時間培養した。前記培養後、ＴＮＦαおよびＩＮＦγを１０ｎｇ／ｍｌ（最終濃度）になるように添加し、１６時間培養した。前記培養後、ＩＬ－３１（１００ｎｇ／ｍｌ）、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍＭカルシウムＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を１時間培養した。前記培養後、前記培養上清を回収し、遠心分離を行い、デブリを除去した。前記除去後、Leukotoriene B4 EIA Kit（520111、Cayman Chemical社製（販売元：富士フイルム和光純薬、513-92091））を用いて、ＥＬＩＳＡを行った。前記ＥＬＩＳＡの結果から、ＬＴＢ４産生量を定量化した。統計解析は、Ｓｔｕｄｅｎｔ’ｓ　ｔ－ｔｅｓｔにより実施した。これらの結果を図６に示す。 (5) Leukotriene B4 Production Test We investigated whether the inhibition of IL-31RA gene expression by Sph1 inhibits the production of leukotriene B4 (LTB4), which causes itching. Specifically, NHEK cells were seeded in a 24-well plate at 1.0×10 5 cells/well. KBM™ medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM™ medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotic (GA-1000), and 0.06 mM calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, 0.5 μg/ml or 1 μg/ml of sphingoid bases (Sph1) were added and cultured for 1 hour. After the culture, TNFα and INFγ were added to a final concentration of 10 ng/ml, and the cells were cultured for 16 hours. After the culture, the medium was replaced with IL-31 (100 ng/ml), BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotics (GA-1000), and 0.06 mM calcium KBM™ medium. After the replacement, the NHEK cells were cultured for 1 hour. After the culture, the culture supernatant was collected and centrifuged to remove debris. After the removal, ELISA was performed using Leukotoriene B4 EIA Kit (520111, Cayman Chemical (distributor: Fujifilm Wako Pure Chemical, 513-92091)). The amount of LTB4 produced was quantified from the results of the ELISA. Statistical analysis was performed using Student's t-test. These results are shown in FIG. 6.
　図６は、ＮＨＥＫ細胞における、ＬＴＢ４産生量を示すグラフである。図６において、縦軸は、ＬＴＢ４産生量（ｐｇ／ｍｌ）を示し、横軸は、試料の種類を示す。
　図６における縦軸の数値は、細胞数で補正したＬＴＢ４産生量として、各群の値を示している。
　図６の縦軸の数値は、以下の通りである。
・Control群：31.65、No add群：47.60、0.5μg/ml Sph1群：30.42、1μg/ml Sph1群：26.68
 
　図６に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＬＴＢ４の産生量が有意に抑制された。以上のことから、表皮角化細胞において、Ｓｐｈ１は、ＬＴＢ４産生を抑制することがわかった。これらの結果から、Ｓｐｈ１が、ＩＬ－３１ＲＡの発現を抑制することにより、ＩＬ－３１を介したＬＴＢ４の産生を抑制し、これにより掻痒を抑制することが示唆された。 Fig. 6 is a graph showing the amount of LTB4 produced in NHEK cells, in which the vertical axis indicates the amount of LTB4 produced (pg/ml) and the horizontal axis indicates the type of sample.
The values on the vertical axis in FIG. 6 indicate the values for each group as the amount of LTB4 produced corrected by the cell number.
The values on the vertical axis in FIG.
Control group: 31.65, No add group: 47.60, 0.5μg/ml Sph1 group: 30.42, 1μg/ml Sph1 group: 26.68

As shown in Figure 6, the amount of LTB4 production was significantly suppressed in the Sph1 group compared to the No add (negative control) group. From the above, it was found that Sph1 suppresses LTB4 production in epidermal keratinocytes. These results suggest that Sph1 suppresses IL-31-mediated LTB4 production by suppressing IL-31RA expression, thereby suppressing pruritus.
　なお、上述の実施例１（２）、実施例１（３）、および実施例１（４）で用いたプライマーの配列等情報を下記表１にも示す。 The sequences and other information of the primers used in the above-mentioned Examples 1(2), 1(3), and 1(4) are also shown in Table 1 below.
［実施例２］
　コメ由来グルコシルセラミドに含まれるグルコシルセラミドの構造タイプを分析し、分析結果からグルコシルセラミドに含有されるスフィンゴイド塩基種の構成比率を推察した。さらに、前記コメ由来グルコシルセラミドに含まれるスフィンゴイド塩基が、掻痒抑制効果を発揮するかどうかを確認した。 [Example 2]
The structural type of glucosylceramide contained in rice-derived glucosylceramide was analyzed, and the composition ratio of sphingoid base species contained in glucosylceramide was inferred from the analysis results.Furthermore, it was confirmed whether the sphingoid base contained in the rice-derived glucosylceramide exhibits an antipruritic effect.
（１）グルコシルセラミドの構造の解析
　コメ由来のグルコシルセラミドに含まれるグルコシルセラミドの構造タイプを解析し、前記コメ由来のグルコシルセラミドに含まれるスフィンゴイド塩基のタイプを推察した。具体的には、１００ｍｇのコメ由来のグルコシルセラミド（フィトセラ、NW-10、Lot No.121520、一丸ファルコス社製）を３ｍｌの精製水に添加し、超音波処理を行い、溶解させた。前記溶解後、１８ｍｌのメタノールを徐々に添加し、超音波処理を行った。その後、放冷し、全体が５０ｍｌになるようにクロロホルムを添加し、抽出を行った。前記抽出後、抽出液をろ過し、ろ液の濃縮乾固を行った。前記濃縮乾固後、５０ｍｌのメタノールを添加し、溶解させた。前記溶解後、再び濃縮乾固を行った。その後、全体が５０ｍｌになるようにメタノールを添加し、０．４５μｍのメンブレンフィルターを用いてろ過した。前記ろ過で得たろ液を検体として用いた。前記ろ液および下記表２に示す標準試薬（長良サイエンス株式会社製）を用いて、液体クロマトグラフィーを行い、各グルコシルセラミド量を測定した。前記液体クロマトグラフィーは、下記表３に示す条件で行った。これらの結果を、下記表４および表５に示す。 (1) Analysis of glucosylceramide structure The structural type of glucosylceramide contained in rice-derived glucosylceramide was analyzed, and the type of sphingoid base contained in the rice-derived glucosylceramide was inferred. Specifically, 100 mg of rice-derived glucosylceramide (Phytocera, NW-10, Lot No. 121520, Ichimaru Falcos) was added to 3 ml of purified water, and sonicated to dissolve. After the dissolution, 18 ml of methanol was gradually added and sonicated. Then, the mixture was allowed to cool, and chloroform was added to a total volume of 50 ml to perform extraction. After the extraction, the extract was filtered, and the filtrate was concentrated and dried. After the concentration and drying, 50 ml of methanol was added and dissolved. After the dissolution, the mixture was concentrated and dried again. Then, methanol was added to a total volume of 50 ml, and filtered using a 0.45 μm membrane filter. The filtrate obtained by the filtration was used as a specimen. The filtrate and the standard reagents (manufactured by Nagara Science Co., Ltd.) shown in Table 2 below were used to perform liquid chromatography to measure the amount of each glucosylceramide. The liquid chromatography was performed under the conditions shown in Table 3 below. The results are shown in Tables 4 and 5 below.
　前記表４および表５に示すように、コメ由来グルコシルセラミドは、８１％のスフィンガジエニン（ｄ１８：２）（Sph1）および１２．６％のスフィンゲニン（ｔ１８：１）（Sph2）を含むことがわかった。以上のことから、コメ由来のグルコシルセラミド（フィトセラ、NW-10、Lot No.121520、一丸ファルコス社製）は、スフィンガジエニンを主に含有することがわかった。 As shown in Tables 4 and 5, rice-derived glucosylceramide was found to contain 81% sphingadienine (d18:2) (Sph1) and 12.6% sphingenine (t18:1) (Sph2). From the above, it was found that rice-derived glucosylceramide (Phytosera, NW-10, Lot No. 121520, Ichimaru Pharcos) mainly contains sphingadienine.
（２）ＴＳＬＰの遺伝子発現量の試験
　スフィンガジエニンを主に含有するコメ由来グルコシルセラミドのスフィンゴイド塩基が、掻痒のバイオマーカーの一種であるＴＳＬＰの遺伝子の発現を抑制するか検討した。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ヒト角化細胞であるＮＨＥＫ細胞（KK-4009、凍結NHEK(NB)、新生児由来、クラボウ社製）を播種した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍｍｏｌ／ｌ　カルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、０．１μｇ／ｍｌ、０．５μｇ／ｍｌ、または１μｇ／ｍｌの濃度のＳｐｈ１またはコメ由来グルコシルセラミド（ｔＳｐｈ）を添加し、１時間培養した。前記培養後、刺激因子であるＴＮＦαを１ｎｇ／ｍｌ（最終濃度）およびＩＮＦγを１ｎｇ／ｍｌ（最終濃度）となるように添加し、６時間培養した。前記培養後、前記実施例１（２）と同様にして、ｑＲＴ－ＰＣＲおよび解析を行った。これらの結果を図７に示す。 (2) Test of TSLP gene expression level We investigated whether the sphingoid base of rice-derived glucosylceramide, which mainly contains sphingadienine, suppresses the expression of the gene for TSLP, which is a type of biomarker for pruritus. Specifically, human keratinocytes, NHEK cells (KK-4009, frozen NHEK (NB), derived from newborns, manufactured by Kurabo Industries, Ltd.), were seeded on a 24-well plate at 1.0 x 10 5 cells/well. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM (trademark) medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotics (GA-1000), and 0.06 mmol/l calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, Sph1 or rice-derived glucosylceramide (tSph) was added at a concentration of 0.1 μg/ml, 0.5 μg/ml, or 1 μg/ml, and the cells were cultured for 1 hour. After the culture, the stimulatory factors TNFα and INFγ were added at 1 ng/ml (final concentration) and 1 ng/ml (final concentration), respectively, and the cells were cultured for 6 hours. After the culture, qRT-PCR and analysis were performed in the same manner as in Example 1(2). The results are shown in FIG. 7.
　図７は、ＮＨＥＫ細胞における、ＴＳＬＰの遺伝子発現量を示すグラフである。図７（Ａ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図７（Ｂ）において、縦軸は、ＴＳＬＰ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。
　図７（Ａ）および図７（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図７（Ａ）の縦軸の数値は、以下の通りである。
・No add群：7.31、0.1μg/ml Sph1群：4.37、0.5μg/ml Sph1群：3.98、1μg/ml Sph1群：3.87
　図７（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：7.31、0.1μg/ml tSph群：4.02、0.5μg/ml tSph群：4.79、1μg/ml tSph群：3.59
 
　図７（Ａ）に示すように、Ｓｐｈ１群では、いずれの濃度でも、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量は有意に抑制された。また、図７（Ｂ）に示すように、ｔＳｐｈ群では、いずれの濃度でも、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＴＳＬＰの遺伝子発現量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１だけでなく、ｔＳｐｈも、ＴＳＬＰの遺伝子発現を抑制することがわかった。 7 is a graph showing the expression level of TSLP gene in NHEK cells. In Fig. 7(A), the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample. In Fig. 7(B), the vertical axis shows the relative value of the expression level of TSLP gene, and the horizontal axis shows the type of sample.
The numerical values on the vertical axis in Figures 7(A) and 7(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 7.31, 0.1 μg/ml Sph1 group: 4.37, 0.5 μg/ml Sph1 group: 3.98, 1 μg/ml Sph1 group: 3.87
The values on the vertical axis in FIG.
No add group: 7.31, 0.1 μg/ml tSph group: 4.02, 0.5 μg/ml tSph group: 4.79, 1 μg/ml tSph group: 3.59

As shown in Figure 7(A), in the Sph1 group, the gene expression level of TSLP was significantly suppressed at all concentrations compared to the No add (negative control) group. Also, as shown in Figure 7(B), in the tSph group, the gene expression level of TSLP was significantly suppressed at all concentrations compared to the No add (negative control) group. From the above, it was found that not only Sph1 but also tSph suppresses the gene expression of TSLP in keratinocytes.
（３）ＩＬ－３１ＲＡの遺伝子発現量の試験
　スフィンガジエニンを主に含有するコメ由来グルコシルセラミドのスフィンゴイド塩基が、ＴＳＬＰの遺伝子発現の抑制に加えて、ＩＬ－３１ＲＡの遺伝子発現の抑制に寄与するか検討した。具体的には、細胞にＮＨＥＫ細胞（KK-4009、凍結NHEK(NB)、新生児由来、クラボウ社製）を使用したこと、セラミドとして０．１μｇ／ｍｌ、０．５μｇ／ｍｌ、または１μｇ／ｍｌの濃度のＳｐｈ１またはコメ由来グルコシルセラミド（ｔＳｐｈ）を使用したこと、および、サイトカインとしてＴＮＦαおよびＩＮＦγを１ｎｇ／ｍｌ（最終濃度）で使用した以外は、前記実施例１（４）と同様にして、ｑＲＴ－ＰＣＲおよび解析を行った。これらの結果を図８に示す。 (3) Test of IL-31RA gene expression level We investigated whether the sphingoid base of rice-derived glucosylceramide, which mainly contains sphingadienine, contributes to the suppression of IL-31RA gene expression in addition to the suppression of TSLP gene expression. Specifically, qRT-PCR and analysis were performed in the same manner as in Example 1(4) above, except that NHEK cells (KK-4009, frozen NHEK (NB), derived from a newborn, manufactured by Kurabo) were used as cells, Sph1 or rice-derived glucosylceramide (tSph) at a concentration of 0.1 μg/ml, 0.5 μg/ml, or 1 μg/ml was used as ceramide, and TNFα and INFγ were used at 1 ng/ml (final concentration) as cytokines. These results are shown in FIG. 8.
　図８は、ＮＨＥＫ細胞における、ＩＬ－３１ＲＡの遺伝子発現量を示すグラフである。図８（Ａ）において、縦軸は、ＩＬ－３１ＲＡ遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図８（Ｂ）において、縦軸は、ＩＬ－３１ＲＡ遺伝子の発現量の相対値を示し、横軸は、試料の種類を示す。
 
　図８（Ａ）および図８（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図８（Ａ）の縦軸の数値は、以下の通りである。
・No add群：9.50、0.1μg/ml Sph1群：5.84、0.5μg/ml Sph1群：5.04、1μg/ml Sph1群：4.55
　図８（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：9.50、0.1μg/ml tSph群：5.74、0.5μg/ml tSph群：5.67、1μg/ml tSph群：4.09
 
　図８（Ａ）に示すように、Ｓｐｈ１群では、いずれの濃度でも、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＩＬ－３１ＲＡの遺伝子発現量は有意に抑制された。また、図８（Ｂ）に示すように、ｔＳｐｈ群では、いずれの濃度でも、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ＩＬ－３１ＲＡの遺伝子発現量が有意に抑制された。以上のことから、角化細胞において、Ｓｐｈ１だけでなく、ｔＳｐｈも、ＩＬ－３１ＲＡの遺伝子発現を抑制することがわかった。 Fig. 8 is a graph showing the gene expression level of IL-31RA in NHEK cells. In Fig. 8(A), the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample. In Fig. 8(B), the vertical axis shows the relative value of the expression level of IL-31RA gene, and the horizontal axis shows the type of sample.

The numerical values on the vertical axis in FIG. 8(A) and FIG. 8(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 9.50, 0.1 μg/ml Sph1 group: 5.84, 0.5 μg/ml Sph1 group: 5.04, 1 μg/ml Sph1 group: 4.55
The values on the vertical axis in FIG.
No add group: 9.50, 0.1 μg/ml tSph group: 5.74, 0.5 μg/ml tSph group: 5.67, 1 μg/ml tSph group: 4.09

As shown in Figure 8(A), in the Sph1 group, the gene expression level of IL-31RA was significantly suppressed at all concentrations compared to the No add (negative control) group. Also, as shown in Figure 8(B), in the tSph group, the gene expression level of IL-31RA was significantly suppressed at all concentrations compared to the No add (negative control) group. From the above, it was found that not only Sph1 but also tSph suppresses the gene expression of IL-31RA in keratinocytes.
（４）ＩＬ－８の遺伝子発現量の試験
　Ｓｐｈ１、およびスフィンガジエニンを主に含有するコメ由来グルコシルセラミドのスフィンゴイド塩基が、炎症のバイオマーカーの一種であるＩＬ－８の遺伝子の発現を抑制するか検討した。具体的には、２４ウェルプレートに、１．０×１０５ cells/wellとなるように、ＮＨＥＫ細胞（KK-4009、凍結NHEK(NB)、新生児由来、クラボウ社製）を播種した。培地は、ＫＢＭ（商標）培地を使用した。前記播種後、前記ＮＨＥＫ細胞がコンフルエントになるまで培養した。前記培養後、各ウェルの培地を、ＢＰＥ（ウシ下垂体抽出液）、インスリン、ＥＧＦ（上皮成長因子）、抗生剤（ＧＡ－１０００）、および０．０６ｍｍｏｌ／ｌ　カルシウムを含有したＫＢＭ（商標）培地に交換した。前記交換後、前記ＮＨＥＫ細胞を２４時間培養した。前記培養後、０．５μｇ／ｍｌまたは１μｇ／ｍｌの濃度のＳｐｈ１、または１μｇ／ｍｌのコメ由来グルコシルセラミド（ｔＳｐｈ）を添加し、１時間培養した。前記培養後、ＴＮＦαおよびＩＮＦγを１ｎｇ／ｍｌ（最終濃度）になるように添加し、６時間培養した。前記培養後、前記培養後、培養上清を回収し、RNeasy Mini Kit（QIAGEN社製）を用いて、添付のプロトコルに従ってｍＲＮＡを精製した。前記精製後、PrimeScript Master Mix（TaKaRa社製）を用いて逆転写反応を行った。つぎに、TB Green Master Mix（TaKaRa社製）記載の方法に従い、下記ＩＬ－８遺伝子用プライマーセットおよびＲＰＳ遺伝子用プライマーセットを用いて、ｑＲＴ－ＰＣＲを行った。ＰＣＲ装置には、LightCycler（登録商標）96（Roche社製）を用いた。ＩＬ－８遺伝子の発現量は、内部標準遺伝子（ＲＰＳ１８遺伝子）の発現量に対する相対値とした。データの解析は、付属のソフトウェアを用いて解析した。統計解析は、ＪＭＰ８を用い、Ｔｕｒｋｙ検定およびＤｕｎｎｅｔｔ’ｓ　ｔｅｓｔにより実施した。これらの結果を図９に示す。 (4) Test of IL-8 gene expression level We investigated whether the sphingoid base of rice-derived glucosylceramide, which mainly contains Sph1 and sphingadienine, suppresses the expression of the IL-8 gene, which is a type of biomarker for inflammation. Specifically, NHEK cells (KK-4009, frozen NHEK (NB), derived from a newborn, manufactured by Kurabo Industries, Ltd.) were seeded on a 24-well plate at 1.0 x 10 5 cells/well. KBM (trademark) medium was used as the medium. After the seeding, the NHEK cells were cultured until they became confluent. After the culture, the medium in each well was replaced with KBM (trademark) medium containing BPE (bovine pituitary extract), insulin, EGF (epidermal growth factor), antibiotic (GA-1000), and 0.06 mmol/l calcium. After the replacement, the NHEK cells were cultured for 24 hours. After the culture, Sph1 at a concentration of 0.5 μg/ml or 1 μg/ml, or rice-derived glucosylceramide (tSph) at 1 μg/ml was added and cultured for 1 hour. After the culture, TNFα and INFγ were added to a final concentration of 1 ng/ml and cultured for 6 hours. After the culture, the culture supernatant was collected, and mRNA was purified using an RNeasy Mini Kit (manufactured by QIAGEN) according to the attached protocol. After the purification, reverse transcription was performed using PrimeScript Master Mix (manufactured by TaKaRa). Next, qRT-PCR was performed using the following primer set for the IL-8 gene and primer set for the RPS gene according to the method described in TB Green Master Mix (manufactured by TaKaRa). The PCR device used was LightCycler (registered trademark) 96 (manufactured by Roche). The expression level of the IL-8 gene was taken as a relative value to the expression level of the internal standard gene (RPS18 gene). Data analysis was performed using the attached software. Statistical analysis was performed using Turky's test and Dunnett's test with JMP 8. The results are shown in FIG.
ＩＬ－８遺伝子用プライマーセット
　フォワードプライマー（配列番号７）
　　5’-AAATTGAGGCCAAGGGCCA-3’
　リバースプライマー（配列番号８）
　　5’-AACCAAGGCACAGTGGAACA-3’ Primer set for IL-8 gene Forward primer (SEQ ID NO: 7)
5'-AAATTGAGGCCAAGGGCCA-3'
Reverse primer (SEQ ID NO:8)
5'-AACCAAGGCACAGTGGAACA-3'
　図９は、ＮＨＥＫ細胞における、ＩＬ－８の遺伝子発現量を示すグラフである。図９（Ａ）において、縦軸は、ＩＬ－８遺伝子発現量の相対値を示し、横軸は、試料の種類を示す。図９（Ｂ）において、縦軸は、ＩＬ－８遺伝子の発現量の相対値を示し、横軸は、試料の種類を示す。
 
　図９（Ａ）および図９（Ｂ）における縦軸の数値は、control群の値を1.0（1.00）として、各群の値を示している。
　図９（Ａ）の縦軸の数値は、以下の通りである。
・No add群：110.61、0.5μg/ml Sph1群：64.96、1μg/ml Sph1群：47.49
　図９（Ｂ）の縦軸の数値は、以下の通りである。
・No add群：110.61、1μg/ml tSph群：58.25
 
　図９（Ａ）に示すように、Ｓｐｈ１群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、Ｓｐｈ１を０．５μｇ／ｍｌまたは１μｇ／ｍｌの濃度で添加した場合、ＩＬ－８の遺伝子発現量は有意に抑制された。また、図９（Ｂ）に示すように、ｔＳｐｈ群では、Ｎｏ　ａｄｄ（ネガティブコントロール）群と比べ、ｔＳｐｈを１μｇ／ｍｌの濃度で添加した場合、ＩＬ－８の遺伝子発現量が有意に抑制された。これらの結果から、角化細胞において、Ｓｐｈ１およびｔＳｐｈは、ＩＬ－８の遺伝子発現を抑制すること、すなわち炎症を抑制することがわかった。 Fig. 9 is a graph showing the expression level of IL-8 gene in NHEK cells. In Fig. 9(A), the vertical axis shows the relative value of the expression level of IL-8 gene, and the horizontal axis shows the type of sample. In Fig. 9(B), the vertical axis shows the relative value of the expression level of IL-8 gene, and the horizontal axis shows the type of sample.

The numerical values on the vertical axis in FIG. 9(A) and FIG. 9(B) indicate the values of each group, with the value of the control group being 1.0 (1.00).
The values on the vertical axis in FIG.
No add group: 110.61, 0.5 μg/ml Sph1 group: 64.96, 1 μg/ml Sph1 group: 47.49
The values on the vertical axis in FIG.
No add group: 110.61, 1μg/ml tSph group: 58.25

As shown in Figure 9 (A), in the Sph1 group, the gene expression level of IL-8 was significantly suppressed when Sph1 was added at a concentration of 0.5 μg/ml or 1 μg/ml, compared to the No add (negative control) group. Also, as shown in Figure 9 (B), in the tSph group, the gene expression level of IL-8 was significantly suppressed when tSph was added at a concentration of 1 μg/ml, compared to the No add (negative control) group. From these results, it was found that Sph1 and tSph suppress the gene expression of IL-8 in keratinocytes, i.e., suppress inflammation.
　なお、上述の実施例２（４）で用いたプライマーの配列等情報を下記表６にも示す。 The sequences and other information of the primers used in Example 2(4) above are also shown in Table 6 below.
　以上、実施形態および実施例を参照して本開示を説明したが、本開示は、上記実施形態および実施例に限定されるものではない。本開示の構成や詳細には、本開示のスコープ内で当業者が理解しうる様々な変更をすることができる。 The present disclosure has been described above with reference to embodiments and examples, but the present disclosure is not limited to the above embodiments and examples. Various modifications that can be understood by a person skilled in the art can be made to the configuration and details of the present disclosure within the scope of the present disclosure.
　本明細書において引用した特許、特許出願および文献は、その内容自体が具体的に本明細書に記載されているのと同様にその内容が本明細書に対する参考として援用される。 　The patents, patent applications, and literature cited in this specification are incorporated by reference into this specification in the same manner as if the contents were specifically set forth in this specification.
　この出願は、２０２２年１１月１１日に出願された日本出願特願２０２２－１８１３２７を基礎とする優先権を主張し、その開示の全てをここに取り込む。 This application claims priority based on Japanese Patent Application No. 2022-181327, filed on November 11, 2022, the entire disclosure of which is incorporated herein by reference.
＜付記＞
　上記の実施形態および実施例の一部または全部は、以下の付記のように記載されうるが、以下には限られない。
＜スフィンゴイド塩基含む掻痒抑制に用いるための剤＞
（付記１）
スフィンゴイド塩基を含む、掻痒抑制に用いるための剤。
（付記２）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１に記載の剤。
（付記３）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１に記載の剤。
（付記４）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、掻痒抑制に用いるための剤。
（付記５）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記４に記載の剤。
（付記６）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記４または５に記載の剤。
（付記７）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記４または５に記載の剤。
（付記８）
前記セラミドは、さらに、糖部を含む、付記４から７のいずれかに記載の剤。
（付記９）
前記糖部は、グルコースに由来する、付記８に記載の剤。
（付記１０）
付記１から９のいずれかに記載の剤を含む、皮膚に塗布するための剤。
（付記１１）
付記１から９のいずれかに記載の剤を含む、経口投与に用いるための剤。
＜スフィンゴイド塩基を含む掻痒抑制に用いるための組成物＞
（付記１２）
スフィンゴイド塩基を含む、掻痒抑制に用いるための組成物。
（付記１３）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１２に記載の組成物。
（付記１４）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１２に記載の組成物。
（付記１５）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、掻痒抑制に用いるための組成物。
（付記１６）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記１５に記載の組成物。
（付記１７）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記１５または１６に記載の組成物。
（付記１８）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記１５または１６に記載の組成物。
（付記１９）
前記セラミドは、さらに、糖部を含む、付記１５から１８のいずれかに記載の組成物。
（付記２０）
前記糖部は、グルコースに由来する、付記１９に記載の組成物。
（付記２１）
付記１２から２０のいずれかに記載の組成物を含む、皮膚に塗布するための組成物。
（付記２２）
付記１２から２０のいずれかに記載の組成物を含む、経口投与に用いるための組成物。
（付記２３）
付記１から１１のいずれかに記載の掻痒抑制に用いるための剤を含む、付記１２から２２のいずれかに記載の組成物。
＜ＴＡＲＣ遺伝子の発現抑制に用いるための剤＞
（付記２４）
スフィンゴイド塩基を含む、ＴＡＲＣ（Thymus and Activation-Regulated Chemokine）遺伝子の発現の抑制に用いるための剤。
（付記２５）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記２４に記載の剤。
（付記２６）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記２４に記載の剤。
（付記２７）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＡＲＣ遺伝子の発現の抑制に用いるための剤。
（付記２８）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記２７に記載の剤。
（付記２９）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記２７または２８に記載の剤。
（付記３０）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記２７または２８に記載の剤。
（付記３１）
前記セラミドは、さらに、糖部を含む、付記２７から３０のいずれかに記載の剤。
（付記３２）
前記糖部は、グルコースに由来する、付記３１に記載の剤。
（付記３３）
付記２４から３２のいずれかに記載の剤を含む、皮膚に塗布するための剤。
（付記３４）
付記２４から３２のいずれかに記載の剤を含む、経口投与に用いるための剤。
＜ＴＡＲＣ遺伝子の発現抑制に用いるための組成物＞
（付記３５）
スフィンゴイド塩基を含む、ＴＡＲＣ遺伝子の発現の抑制に用いるための組成物。
（付記３６）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記３５に記載の組成物。
（付記３７）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記３５に記載の組成物。
（付記３８）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＡＲＣ遺伝子の発現の抑制に用いるための組成物。
（付記３９）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記３８記載の組成物。
（付記４０）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記３８または３９に記載の組成物。
（付記４１）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記３８または３９に記載の組成物。
（付記４２）
前記セラミドは、さらに、糖部を含む、付記３８から４１のいずれかに記載の組成物。
（付記４３）
前記糖部は、グルコースに由来する、付記４２に記載の組成物。
（付記４４）
付記３５から４３のいずれかに記載の組成物を含む、皮膚に塗布するための組成物。
（付記４５）
付記３５から４３のいずれかに記載の組成物を含む、経口投与に用いるための組成物。
（付記４６）
付記２４から３４のいずれかに記載のＴＡＲＣ遺伝子の発現の抑制に用いるための剤を含む、付記３５から４５のいずれかに記載の組成物。
＜ＴＳＬＰ遺伝子の発現抑制に用いるための剤＞
（付記４７）
スフィンゴイド塩基を含む、ＴＳＬＰ（Thymic stromal lymphopoietin）遺伝子の発現の抑制に用いるための剤。
（付記４８）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記４７に記載の剤。
（付記４９）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記４７に記載の剤。
（付記５０）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＳＬＰ遺伝子の発現の抑制に用いるための剤。
（付記５１）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記５０に記載の剤。
（付記５２）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記５０または５１に記載の剤。
（付記５３）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記５０または５１に記載の剤。
（付記５４）
前記セラミドは、さらに、糖部を含む、付記５０から５３のいずれかに記載の剤。
（付記５５）
前記糖部は、グルコースに由来する、付記５４に記載の剤。
（付記５６）
付記４７から５５のいずれかに記載の剤を含む、皮膚に塗布するための剤。
（付記５７）
付記４７から５５のいずれかに記載の剤を含む、経口投与に用いるための剤。
＜ＴＳＬＰ遺伝子の発現抑制に用いるための組成物＞
（付記５８）
スフィンゴイド塩基を含む、ＴＳＬＰ遺伝子の発現の抑制に用いるための組成物。
（付記５９）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記５８に記載の組成物。
（付記６０）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記５８に記載の組成物。
（付記６１）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＳＬＰ遺伝子の発現の抑制に用いるための組成物。
（付記６２）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記６１に記載の組成物。
（付記６３）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記６１または６２に記載の組成物。
（付記６４）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記６１または６２に記載の組成物。
（付記６５）
前記セラミドは、さらに、糖部を含む、付記６１から６４のいずれかに記載の組成物。
（付記６６）
前記糖部は、グルコースに由来する、付記６５に記載の組成物。
（付記６７）
付記５８から６６のいずれかに記載の組成物を含む、皮膚に塗布するための組成物。
（付記６８）
付記５８から６６のいずれかに記載の組成物を含む、経口投与に用いるための組成物。
（付記６９）
付記４７から５７のいずれかに記載のＴＳＬＰ遺伝子の発現の抑制に用いるための剤を含む、付記５８から６８のいずれかに記載の組成物。
＜ＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための剤＞
（付記７０）
スフィンゴイド塩基を含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤。
（付記７１）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記７０に記載の剤。
（付記７２）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記７０に記載の剤。
（付記７３）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤。
（付記７４）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記７３に記載の剤。
（付記７５）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記７３または７４に記載の剤。
（付記７６）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記７３または７４に記載の剤。
（付記７７）
前記セラミドは、さらに、糖部を含む、付記７３から７６のいずれかに記載の剤。
（付記７８）
前記糖部は、グルコースに由来する、付記７７に記載の剤。
（付記７９）
付記７０から７８のいずれかに記載の剤を含む、皮膚に塗布するための剤。
（付記８０）
付記７０から７８のいずれかに記載の剤を含む、経口投与に用いるための剤。
＜ＩＬ－３１ＲＡ遺伝子の発現抑制に用いるための組成物＞
（付記８１）
スフィンゴイド塩基を含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物。
（付記８２）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記８１に記載の組成物。
（付記８３）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記８１に記載の組成物。
（付記８４）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物。
（付記８５）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記８４に記載の組成物。
（付記８６）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記８４または８５に記載の組成物。
（付記８７）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記８４または８５に記載の組成物。
（付記８８）
前記セラミドは、さらに、糖部を含む、付記８４から８７のいずれかに記載の組成物。
（付記８９）
前記糖部は、グルコースに由来する、付記８８に記載の組成物。
（付記９０）
付記８１から８９のいずれかに記載の組成物を含む、皮膚に塗布するための組成物。
（付記９１）
付記８１から８９のいずれかに記載の組成物を含む、経口投与に用いるための組成物。
（付記９２）
付記７０から８０のいずれかに記載のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤を含む、付記８１から９１のいずれかに記載の組成物。
＜ＩＬ－８遺伝子の発現抑制に用いるための剤＞
（付記９３）
スフィンゴイド塩基を含む、ＩＬ－８遺伝子の発現の抑制に用いるための剤。
（付記９４）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記９３に記載の剤。
（付記９５）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記９３に記載の剤。
（付記９６）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＩＬ－８遺伝子の発現の抑制に用いるための剤。
（付記９７）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記９６に記載の剤。
（付記９８）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記９６または９７に記載の剤。
（付記９９）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記９６または９７に記載の剤。
（付記１００）
前記セラミドは、さらに、糖部を含む、付記９６から９９のいずれかに記載の剤。
（付記１０１）
前記糖部は、グルコースに由来する、付記１００に記載の剤。
（付記１０２）
付記９３から１０１のいずれかに記載の剤を含む、皮膚に塗布するための剤。
（付記１０３）
付記９３から１０１のいずれかに記載の剤を含む、経口投与に用いるための剤。
＜ＩＬ－８遺伝子の発現抑制に用いるための組成物＞
（付記１０４）
スフィンゴイド塩基を含む、ＩＬ－８遺伝子の発現の抑制に用いるための組成物。
（付記１０５）
前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１０４に記載の組成物。
（付記１０６）
前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、付記１０４に記載の組成物。
（付記１０７）
スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＩＬ－８遺伝子の発現の抑制に用いるための組成物。
（付記１０８）
前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、付記１０７に記載の組成物。
（付記１０９）
前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記１０７または１０８に記載の組成物。
（付記１１０）
前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、付記１０７または１０８に記載の組成物。
（付記１１１）
前記セラミドは、さらに、糖部を含む、付記１０７から１１０のいずれかに記載の組成物。
（付記１１２）
前記糖部は、グルコースに由来する、付記１１１に記載の組成物。
（付記１１３）
付記１０４から１１２のいずれかに記載の組成物を含む、皮膚に塗布するための組成物。
（付記１１４）
付記１０４から１１２のいずれかに記載の組成物を含む、経口投与に用いるための組成物。
（付記１１５）
付記９３から１０３のいずれかに記載のＩＬ－８遺伝子の発現の抑制に用いるための剤を含む、付記１０４から１１４のいずれかに記載の組成物。
＜掻痒抑制方法＞
（付記１１６）
付記１から１１のいずれかに記載の掻痒抑制に用いるための剤、および／または、付記１２から２３のいずれかに記載の掻痒抑制に用いるための組成物を使用する、掻痒抑制方法。
（付記１１７）
対象に、前記掻痒抑制に用いるための剤、および／または、前記掻痒抑制に用いるための組成物を使用する使用工程を含む、付記１１６に記載の抑制方法。
（付記１１８）
ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで使用する、付記１１６または１１７に記載の抑制方法。
＜ＴＡＲＣ遺伝子の発現抑制方法＞
（付記１１９）
付記２４から３４のいずれかに記載のＴＡＲＣ遺伝子の発現の抑制に用いるための剤、および／または、付記３５から４６のいずれかに記載のＴＡＲＣ遺伝子の発現の抑制に用いるための組成物を使用する、ＴＡＲＣ遺伝子の発現抑制方法。
（付記１２０）
対象に、前記ＴＡＲＣ遺伝子の発現の抑制に用いるための剤、および／または、前記ＴＡＲＣ遺伝子の発現の抑制に用いるための組成物を使用する使用工程を含む、付記１１９に記載の抑制方法。
（付記１２１）
ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで使用する、付記１１９または１２０に記載の抑制方法。
＜ＴＳＬＰ遺伝子の発現抑制方法＞
（付記１２２）
付記４７から５７のいずれかに記載のＴＳＬＰ遺伝子の発現の抑制に用いるための剤、および／または、付記５８から６９のいずれかに記載のＴＳＬＰ遺伝子の発現の抑制に用いるための組成物を使用する、ＴＳＬＰ遺伝子の発現抑制方法。
（付記１２３）
対象に、前記ＴＳＬＰ遺伝子の発現の抑制に用いるための剤、および／または、前記ＴＳＬＰ遺伝子の発現の抑制に用いるための組成物を使用する使用工程を含む、付記１２２に記載の抑制方法。
（付記１２４）
ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで使用する、付記１２２または１２３に記載の抑制方法。
＜ＩＬ－３１ＲＡ遺伝子の発現抑制方法＞
（付記１２５）
付記７０から８０のいずれかに記載のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤、および／または、付記８１から９２のいずれかに記載のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物を使用する、ＩＬ－３１ＲＡ遺伝子の発現抑制方法。
（付記１２６）
対象に、前記ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤、および／または、前記ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物を使用する使用工程を含む、付記１２５に記載の抑制方法。
（付記１２７）
ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで使用する、付記１２５または１２６に記載の抑制方法。
＜ＩＬ－８遺伝子の発現抑制方法＞
（付記１２８）
付記９３から１０３のいずれかに記載のＩＬ－８遺伝子の発現の抑制に用いるための剤、および／または、付記１０４から１１５のいずれかに記載のＩＬ－８遺伝子の発現の抑制に用いるための組成物を使用する、ＩＬ－８遺伝子の発現抑制方法。
（付記１２９）
対象に、前記ＩＬ－８遺伝子の発現の抑制に用いるための剤、および／または、前記ＩＬ－８遺伝子の発現の抑制に用いるための組成物を使用する使用工程を含む、付記１２８に記載の抑制方法。
（付記１３０）
ｉｎ　ｖｉｔｒｏまたはｉｎ　ｖｉｖｏで使用する、付記１２８または１２９に記載の抑制方法。
＜使用＞
（付記１３１）
掻痒抑制に用いるための、付記１から１１のいずれかに記載の掻痒抑制に用いるための剤、および／または、付記１２から２３のいずれかに記載の掻痒抑制に用いるための組成物の使用。
（付記１３２）
ＴＡＲＣ遺伝子の発現の抑制に用いるための、付記２４から３４のいずれかに記載のＴＡＲＣ遺伝子の発現の抑制に用いるための剤、および／または、付記３５から４６のいずれかに記載のＴＡＲＣ遺伝子の発現の抑制に用いるための組成物の使用。
（付記１３３）
ＴＳＬＰ遺伝子の発現の抑制に用いるための、付記４７から５７のいずれかに記載のＴＳＬＰ遺伝子の発現の抑制に用いるための剤、および／または、付記５８から６９のいずれかに記載のＴＳＬＰ遺伝子の発現の抑制に用いるための組成物の使用。
（付記１３４）
ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための、付記７０から８０のいずれかに記載のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤、および／または、付記８１から９２のいずれかに記載のＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物の使用
（付記１３５）
ＩＬ－８遺伝子の発現の抑制に用いるための、付記９３から１０３のいずれかに記載のＩＬ－８遺伝子の発現の抑制に用いるための剤、および／または、付記１０４から１１５のいずれかに記載のＩＬ－８遺伝子の発現の抑制に用いるための組成物の使用 <Additional Notes>
Some or all of the above-described embodiments and examples may be described as follows, but are not limited to the following supplementary notes.
<Agent containing a sphingoid base for use in suppressing pruritus>
(Appendix 1)
An agent for use in suppressing pruritus, comprising a sphingoid base.
(Appendix 2)
The agent according to claim 1, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 3)
The agent described in Appendix 1, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 4)
An agent for use in suppressing pruritus, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 5)
5. The agent of claim 4, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 6)
The agent according to claim 4 or 5, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 7)
The agent according to claim 4 or 5, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 8)
The agent according to any one of claims 4 to 7, wherein the ceramide further comprises a sugar moiety.
(Appendix 9)
The agent according to claim 8, wherein the sugar moiety is derived from glucose.
(Appendix 10)
10. An agent for application to the skin comprising an agent according to any one of claims 1 to 9.
(Appendix 11)
10. An agent for oral administration comprising the agent according to any one of claims 1 to 9.
<Composition for use in suppressing pruritus containing a sphingoid base>
(Appendix 12)
A composition for use in suppressing pruritus, comprising a sphingoid base.
(Appendix 13)
The composition of claim 12, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 14)
The composition of claim 12, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 15)
A composition for use in suppressing pruritus, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 16)
16. The composition of claim 15, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 17)
The composition of claim 15 or 16, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 18)
The composition of claim 15 or 16, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 19)
19. The composition of any one of claims 15 to 18, wherein the ceramide further comprises a sugar moiety.
(Appendix 20)
20. The composition of claim 19, wherein the sugar moiety is derived from glucose.
(Appendix 21)
21. A composition for application to the skin comprising a composition according to any one of claims 12 to 20.
(Appendix 22)
21. A composition for oral administration comprising a composition according to any one of claims 12 to 20.
(Appendix 23)
A composition described in any one of appendices 12 to 22, comprising an agent for use in suppressing pruritus described in any one of appendices 1 to 11.
<Agent for use in suppressing expression of TARC gene>
(Appendix 24)
An agent for use in inhibiting expression of a Thymus and Activation-Regulated Chemokine (TARC) gene, comprising a sphingoid base.
(Appendix 25)
The agent according to claim 24, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 26)
The agent according to claim 24, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 27)
An agent for use in inhibiting expression of the TARC gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 28)
28. The agent of claim 27, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 29)
The agent according to claim 27 or 28, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 30)
The agent according to claim 27 or 28, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 31)
31. The agent according to any one of claims 27 to 30, wherein the ceramide further comprises a sugar moiety.
(Appendix 32)
32. The agent of claim 31, wherein the sugar moiety is derived from glucose.
(Appendix 33)
33. An agent for application to the skin comprising an agent according to any one of claims 24 to 32.
(Appendix 34)
33. An agent for oral administration comprising an agent according to any one of claims 24 to 32.
<Composition for use in suppressing expression of TARC gene>
(Appendix 35)
A composition for use in inhibiting expression of the TARC gene, comprising a sphingoid base.
(Appendix 36)
The composition of claim 35, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 37)
The composition of claim 35, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 38)
A composition for use in inhibiting expression of the TARC gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 39)
39. The composition of claim 38, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 40)
The composition of claim 38 or 39, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 41)
The composition of claim 38 or 39, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 42)
42. The composition of any one of claims 38 to 41, wherein the ceramide further comprises a sugar moiety.
(Appendix 43)
43. The composition of claim 42, wherein the sugar moiety is derived from glucose.
(Appendix 44)
44. A composition for application to the skin comprising a composition according to any one of claims 35 to 43.
(Appendix 45)
A composition for oral administration comprising a composition according to any one of claims 35 to 43.
(Appendix 46)
A composition described in any one of appendices 35 to 45, comprising an agent for use in inhibiting expression of the TARC gene described in any one of appendices 24 to 34.
<Agent for use in inhibiting expression of TSLP gene>
(Appendix 47)
An agent for use in inhibiting expression of thymic stromal lymphopoietin (TSLP) gene, comprising a sphingoid base.
(Appendix 48)
The agent according to claim 47, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 49)
The agent according to claim 47, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 50)
An agent for use in inhibiting expression of the TSLP gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 51)
51. The agent of claim 50, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 52)
52. The agent according to claim 50 or 51, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 53)
52. The agent according to claim 50 or 51, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 54)
54. The agent according to any one of claims 50 to 53, wherein the ceramide further comprises a sugar moiety.
(Appendix 55)
55. The agent of claim 54, wherein the sugar moiety is derived from glucose.
(Appendix 56)
56. An agent for application to the skin comprising an agent according to any one of claims 47 to 55.
(Appendix 57)
56. An agent for oral administration comprising an agent according to any one of claims 47 to 55.
<Composition for use in inhibiting expression of the TSLP gene>
(Appendix 58)
A composition for use in inhibiting expression of the TSLP gene, comprising a sphingoid base.
(Appendix 59)
The composition of claim 58, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 60)
The composition of claim 58, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 61)
A composition for use in inhibiting expression of the TSLP gene, comprising a ceramide comprising a sphingoid base moiety and a fatty acid moiety.
(Appendix 62)
62. The composition of claim 61, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 63)
The composition of claim 61 or 62, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 64)
The composition of claim 61 or 62, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 65)
65. The composition of any one of claims 61 to 64, wherein the ceramide further comprises a sugar moiety.
(Appendix 66)
66. The composition of claim 65, wherein the sugar moiety is derived from glucose.
(Appendix 67)
67. A composition for application to the skin comprising a composition according to any of claims 58 to 66.
(Appendix 68)
67. A composition for oral administration comprising a composition according to any one of claims 58 to 66.
(Appendix 69)
A composition described in any one of appendices 58 to 68, comprising an agent for use in inhibiting the expression of a TSLP gene described in any one of appendices 47 to 57.
<Agent for use in suppressing expression of IL-31RA gene>
(Appendix 70)
An agent for use in suppressing the expression of the IL-31RA gene, comprising a sphingoid base.
(Appendix 71)
The agent according to claim 70, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 72)
The agent according to claim 70, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 73)
An agent for use in suppressing expression of the IL-31RA gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 74)
74. The agent of claim 73, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 75)
The agent according to claim 73 or 74, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 76)
The agent according to claim 73 or 74, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 77)
77. The agent according to any one of claims 73 to 76, wherein the ceramide further comprises a sugar moiety.
(Appendix 78)
78. The agent of claim 77, wherein the sugar moiety is derived from glucose.
(Appendix 79)
80. An agent for application to the skin comprising an agent according to any one of claims 70 to 78.
(Appendix 80)
80. An agent for oral administration comprising an agent according to any one of claims 70 to 78.
<Composition for use in suppressing expression of IL-31RA gene>
(Appendix 81)
A composition for use in suppressing expression of the IL-31RA gene, comprising a sphingoid base.
(Appendix 82)
The composition of claim 81, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 83)
The composition of claim 81, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 84)
A composition for use in suppressing expression of the IL-31RA gene, comprising a ceramide having a sphingoid base moiety and a fatty acid moiety.
(Appendix 85)
85. The composition of claim 84, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 86)
The composition of claim 84 or 85, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 87)
The composition of claim 84 or 85, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 88)
88. The composition of any one of claims 84 to 87, wherein the ceramide further comprises a sugar moiety.
(Appendix 89)
90. The composition of claim 88, wherein the sugar moiety is derived from glucose.
(Appendix 90)
90. A composition for application to the skin comprising a composition according to any one of claims 81 to 89.
(Appendix 91)
90. A composition for oral administration comprising a composition according to any one of claims 81 to 89.
(Appendix 92)
92. The composition according to any one of appendices 81 to 91, comprising an agent for use in suppressing the expression of the IL-31RA gene according to any one of appendices 70 to 80.
<Agent for use in suppressing expression of IL-8 gene>
(Appendix 93)
An agent for use in suppressing the expression of the IL-8 gene, comprising a sphingoid base.
(Appendix 94)
The agent according to claim 93, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 95)
The agent according to claim 93, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 96)
An agent for use in suppressing expression of the IL-8 gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
(Appendix 97)
97. The agent of claim 96, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 98)
The agent according to claim 96 or 97, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 99)
The agent according to claim 96 or 97, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 100)
100. The agent of any one of claims 96 to 99, wherein the ceramide further comprises a sugar moiety.
(Appendix 101)
101. The agent of claim 100, wherein the sugar moiety is derived from glucose.
(Appendix 102)
102. An agent for application to the skin comprising an agent according to any one of claims 93 to 101.
(Appendix 103)
102. An agent for oral administration comprising the agent according to any one of claims 93 to 101.
<Composition for use in suppressing expression of IL-8 gene>
(Appendix 104)
A composition for use in suppressing expression of the IL-8 gene, comprising a sphingoid base.
(Appendix 105)
The composition of claim 104, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 106)
The composition of claim 104, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 107)
A composition for use in suppressing expression of the IL-8 gene, comprising a ceramide having a sphingoid base moiety and a fatty acid moiety.
(Appendix 108)
108. The composition of claim 107, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
(Appendix 109)
The composition of claim 107 or 108, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 110)
The composition of claim 107 or 108, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-Hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
(Appendix 111)
111. The composition of any one of claims 107 to 110, wherein the ceramide further comprises a sugar moiety.
(Appendix 112)
112. The composition of claim 111, wherein the sugar moiety is derived from glucose.
(Appendix 113)
113. A composition for application to the skin comprising the composition of any of claims 104 to 112.
(Appendix 114)
113. A composition for oral administration comprising the composition of any of claims 104 to 112.
(Appendix 115)
A composition according to any one of claims 104 to 114, comprising an agent for use in suppressing expression of an IL-8 gene according to any one of claims 93 to 103.
Method for suppressing pruritus
(Appendix 116)
A method for suppressing pruritus, comprising using an agent for use in suppressing pruritus described in any one of Appendices 1 to 11 and/or a composition for use in suppressing pruritus described in any one of Appendices 12 to 23.
(Appendix 117)
The method for suppressing itch described in Appendix 116, comprising a step of using the agent for use in suppressing itch and/or the composition for use in suppressing itch on a subject.
(Appendix 118)
118. The method of suppression according to claim 116 or 117, for use in vitro or in vivo .
<Method for inhibiting expression of TARC gene>
(Appendix 119)
A method for inhibiting expression of the TARC gene, comprising using an agent for inhibiting expression of the TARC gene described in any of Appendices 24 to 34 and/or a composition for inhibiting expression of the TARC gene described in any of Appendices 35 to 46.
(Appendix 120)
The method for suppressing expression of the TARC gene described in Appendix 119, comprising a step of using an agent for suppressing expression of the TARC gene and/or a composition for suppressing expression of the TARC gene in a subject.
(Appendix 121)
121. The method of suppression according to claim 119 or 120, for use in vitro or in vivo .
<Method for inhibiting expression of TSLP gene>
(Appendix 122)
A method for inhibiting expression of the TSLP gene, comprising using an agent for use in inhibiting expression of the TSLP gene described in any of Appendices 47 to 57, and/or a composition for use in inhibiting expression of the TSLP gene described in any of Appendices 58 to 69.
(Appendix 123)
The method of suppression described in Appendix 122, comprising a step of using an agent for use in suppressing expression of the TSLP gene and/or a composition for use in suppressing expression of the TSLP gene in a subject.
(Appendix 124)
124. The method of suppression according to claim 122 or 123, for use in vitro or in vivo .
<Method for inhibiting expression of IL-31RA gene>
(Appendix 125)
A method for suppressing expression of the IL-31RA gene, comprising using an agent for use in suppressing expression of the IL-31RA gene described in any of Appendices 70 to 80 and/or a composition for use in suppressing expression of the IL-31RA gene described in any of Appendices 81 to 92.
(Appendix 126)
The method for suppressing expression of the IL-31RA gene according to claim 125, comprising a step of using an agent for use in suppressing expression of the IL-31RA gene and/or a composition for use in suppressing expression of the IL-31RA gene in a subject.
(Appendix 127)
127. The method of suppression according to claim 125 or 126, for use in vitro or in vivo .
<Method for inhibiting expression of IL-8 gene>
(Appendix 128)
A method for suppressing expression of the IL-8 gene, comprising using an agent for use in suppressing expression of the IL-8 gene described in any of Appendices 93 to 103 and/or a composition for use in suppressing expression of the IL-8 gene described in any of Appendices 104 to 115.
(Appendix 129)
The method for suppressing expression of the IL-8 gene described in Appendix 128, comprising a step of using an agent for use in suppressing expression of the IL-8 gene and/or a composition for use in suppressing expression of the IL-8 gene in a subject.
(Appendix 130)
130. The method of suppression according to claim 128 or 129, for use in vitro or in vivo .
<Use>
(Appendix 131)
Use of an agent for use in suppressing pruritus described in any one of Appendices 1 to 11 and/or a composition for use in suppressing pruritus described in any one of Appendices 12 to 23 for use in suppressing pruritus.
(Appendix 132)
Use of an agent for use in suppressing expression of the TARC gene described in any of Appendices 24 to 34 and/or a composition for use in suppressing expression of the TARC gene described in any of Appendices 35 to 46.
(Appendix 133)
Use of an agent for use in suppressing expression of the TSLP gene described in any of Appendices 47 to 57 and/or a composition for use in suppressing expression of the TSLP gene described in any of Appendices 58 to 69 for use in suppressing expression of the TSLP gene.
(Appendix 134)
Use of an agent for use in suppressing expression of the IL-31RA gene according to any one of appendices 70 to 80 and/or a composition for use in suppressing expression of the IL-31RA gene according to any one of appendices 81 to 92 (Appendix 135)
Use of an agent for use in suppressing the expression of the IL-8 gene according to any one of appendices 93 to 103 and/or a composition for use in suppressing the expression of the IL-8 gene according to any one of appendices 104 to 115
　以上説明したように、本開示によれば、掻痒関連遺伝子の発現抑制効果を誘導でき、主に、ヒトの皮膚等に適用可能な新たな剤を提供できる。このため、本開示は、例えば、医薬品、医薬部外品、皮膚外用剤等の分野において、極めて有用といえる。 As explained above, the present disclosure can provide a new agent that can induce the effect of suppressing the expression of pruritus-related genes and can be applied primarily to human skin. For this reason, the present disclosure can be said to be extremely useful, for example, in the fields of pharmaceuticals, quasi-drugs, and topical skin preparations.

Claims (48)

  1. スフィンゴイド塩基を含む、掻痒抑制に用いるための剤。 An agent containing a sphingoid base for use in suppressing pruritus.
  2. 前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項１に記載の剤。 The agent according to claim 1, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  3. 前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項１に記載の剤。 The agent according to claim 1, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  4. スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、掻痒抑制に用いるための剤。 An agent for use in suppressing pruritus, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  5. 前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、請求項４に記載の剤。 The agent according to claim 4, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  6. 前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項４または５に記載の剤。 The agent according to claim 4 or 5, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  7. 前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項４または５に記載の剤。 The agent according to claim 4 or 5, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  8. 前記セラミドは、さらに、糖部を含む、請求項４から７のいずれか一項に記載の剤。 The agent according to any one of claims 4 to 7, wherein the ceramide further comprises a sugar moiety.
  9. 前記糖部は、グルコースに由来する、請求項８に記載の剤。 The agent according to claim 8, wherein the sugar portion is derived from glucose.
  10. 請求項１から９のいずれか一項に記載の剤を含む、皮膚に塗布するための剤。 An agent for application to the skin, comprising an agent according to any one of claims 1 to 9.
  11. 請求項１から９のいずれか一項に記載の剤を含む、経口投与に用いるための剤。 A preparation for oral administration comprising the preparation according to any one of claims 1 to 9.
  12. スフィンゴイド塩基を含む、掻痒抑制に用いるための組成物。 A composition for use in suppressing pruritus, comprising a sphingoid base.
  13. スフィンゴイド塩基を含む、ＴＡＲＣ（Thymus and Activation-Regulated Chemokine）遺伝子の発現の抑制に用いるための剤。 An agent containing a sphingoid base for use in inhibiting the expression of the TARC (Thymus and Activation-Regulated Chemokine) gene.
  14. 前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項１３に記載の剤。 The agent according to claim 13, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  15. 前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項１３に記載の剤。 The agent according to claim 13, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  16. スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＡＲＣ遺伝子の発現の抑制に用いるための剤。 An agent for use in suppressing the expression of the TARC gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  17. 前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、請求項１６記載の剤。 The agent according to claim 16, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  18. 前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項１６または１７に記載の剤。 The agent according to claim 16 or 17, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  19. 前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項１６または１７に記載の剤。 The agent according to claim 16 or 17, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  20. 前記セラミドは、さらに、糖部を含む、請求項１６から１９のいずれか一項に記載の剤。 The agent according to any one of claims 16 to 19, wherein the ceramide further comprises a sugar moiety.
  21. 前記糖部は、グルコースに由来する、請求項２０に記載の剤。 The agent according to claim 20, wherein the sugar portion is derived from glucose.
  22. 請求項１３から２１のいずれか一項に記載の剤を含む、皮膚に塗布するための剤。 An agent for application to the skin, comprising an agent according to any one of claims 13 to 21.
  23. 請求項１３から２１のいずれか一項に記載の剤を含む、経口投与に用いるための剤。 A preparation for oral administration comprising the preparation according to any one of claims 13 to 21.
  24. スフィンゴイド塩基を含む、ＴＡＲＣ遺伝子の発現の抑制に用いるための組成物。 A composition for use in inhibiting expression of the TARC gene, comprising a sphingoid base.
  25. スフィンゴイド塩基を含む、ＴＳＬＰ（Thymic stromal lymphopoietin）遺伝子の発現の抑制に用いるための剤。 An agent containing a sphingoid base for use in inhibiting the expression of the TSLP (thymic stromal lymphopoietin) gene.
  26. 前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項２５に記載の剤。 The agent according to claim 25, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  27. 前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項２５に記載の剤。 The agent according to claim 25, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  28. スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＴＳＬＰ遺伝子の発現の抑制に用いるための剤。 An agent for use in suppressing the expression of the TSLP gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  29. 前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、請求項２８に記載の剤。 The agent of claim 28, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  30. 前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項２８または２９に記載の剤。 The agent according to claim 28 or 29, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  31. 前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項２８または２９に記載の剤。 The agent according to claim 28 or 29, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  32. 前記セラミドは、さらに、糖部を含む、請求項２８から３１のいずれか一項に記載の剤。 The agent according to any one of claims 28 to 31, wherein the ceramide further comprises a sugar moiety.
  33. 前記糖部は、グルコースに由来する、請求項３２に記載の剤。 The agent according to claim 32, wherein the sugar portion is derived from glucose.
  34. 請求項２５から３３のいずれか一項に記載の剤を含む、皮膚に塗布するための剤。 An agent for application to the skin, comprising an agent according to any one of claims 25 to 33.
  35. 請求項２５から３３のいずれか一項に記載の剤を含む、経口投与に用いるための剤。 A preparation for oral administration comprising the preparation according to any one of claims 25 to 33.
  36. スフィンゴイド塩基を含む、ＴＳＬＰ遺伝子の発現の抑制に用いるための組成物。 A composition for use in inhibiting expression of the TSLP gene, comprising a sphingoid base.
  37. スフィンゴイド塩基を含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤。 An agent containing a sphingoid base for use in suppressing the expression of the IL-31RA gene.
  38. 前記スフィンゴイド塩基は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項３７に記載の剤。 The agent according to claim 37, wherein the sphingoid base is 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  39. 前記スフィンゴイド塩基は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineである、請求項３７に記載の剤。 The agent according to claim 37, wherein the sphingoid base is trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  40. スフィンゴイド塩基部と、脂肪酸部とを含むセラミドを含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための剤。 An agent for use in suppressing the expression of the IL-31RA gene, comprising a ceramide having a sphingoid base portion and a fatty acid portion.
  41. 前記脂肪酸部は、パルミチン酸（16:0）、ステアリン酸（18:0）、オレイン酸（18:1）、アラキジン酸（20:0）、ベヘン酸（22:0）、エルカ酸（22：1）、トリコシル酸（23:0）、リグノセリン酸（24:0）、またはネルボン酸（24:1）に由来する、請求項４０に記載の剤。 The agent of claim 40, wherein the fatty acid moiety is derived from palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), arachidic acid (20:0), behenic acid (22:0), erucic acid (22:1), tricosylic acid (23:0), lignoceric acid (24:0), or nervonic acid (24:1).
  42. 前記スフィンゴイド塩基部は、4-8-sphingadienine、4-Hydroxy-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項４０または４１に記載の剤。 The agent according to claim 40 or 41, wherein the sphingoid base moiety is derived from 4-8-sphingadienine, 4-hydroxy-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  43. 前記スフィンゴイド塩基部は、trans-4-cis-8-sphingadienine、4-Hydroxy-cis-8-sphingenin、C18-sphingosine、および／またはC20-sphingosineに由来する、請求項４０または４１に記載の剤。 The agent according to claim 40 or 41, wherein the sphingoid base moiety is derived from trans-4-cis-8-sphingadienine, 4-hydroxy-cis-8-sphingenin, C18-sphingosine, and/or C20-sphingosine.
  44. 前記セラミドは、さらに、糖部を含む、請求項４０から４３のいずれか一項に記載の剤。 The agent according to any one of claims 40 to 43, wherein the ceramide further comprises a sugar moiety.
  45. 前記糖部は、グルコースに由来する、請求項４４に記載の剤。 The agent according to claim 44, wherein the sugar portion is derived from glucose.
  46. 請求項３７から４５のいずれか一項に記載の剤を含む、皮膚に塗布するための剤。 An agent for application to the skin, comprising an agent according to any one of claims 37 to 45.
  47. 請求項３７から４５のいずれか一項に記載の剤を含む、経口投与に用いるための剤。 A preparation for oral administration comprising the preparation according to any one of claims 37 to 45.
  48. スフィンゴイド塩基を含む、ＩＬ－３１ＲＡ遺伝子の発現の抑制に用いるための組成物。

          A composition for use in suppressing expression of the IL-31RA gene, comprising a sphingoid base.
PCT/JP2023/022157 2022-11-11 2023-06-14 Agent containing sphingoid base for use in prevention of pruritus, and use thereof WO2024100919A1 (en)

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