Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/13—Amines
  • A61K31/133—Amines having hydroxy groups, e.g. sphingosine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/164—Amides, e.g. hydroxamic acids of a carboxylic acid with an aminoalcohol, e.g. ceramides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
  • A61K31/7032—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a polyol, i.e. compounds having two or more free or esterified hydroxy groups, including the hydroxy group involved in the glycosidic linkage, e.g. monoglucosyldiacylglycerides, lactobionic acid, gangliosides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P17/00—Drugs for dermatological disorders
  • A61P17/04—Antipruritics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• 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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