JP6514980B2 - Itching prevention or remedy - Google Patents

Description

  The present invention relates to an MrpprX1 activity inhibitor, an inhibitor of calcium ion influx into cells via MrpprX1, and an agent for the prevention or improvement of itch.

  Skin diseases such as atopic dermatitis, dry skin and urticaria are accompanied by one of the cutaneous sensations, itching. Such itching is temporarily eliminated by scratching the skin. However, scratching the skin may form secondary skin lesions. Therefore, it is clinically important to prevent or improve itching and to control it.

Histamine is known as a stimulant substance in vivo. When histamine is released in vivo to bind to the histamine receptor, the histamine receptor is activated. Histamine receptors are expressed on sensory nerves such as C fibers that recognize itching. And, when histamine receptors are activated by histamine, itching of the skin is caused via sensory nerves.
Antihistamines are widely used to treat such itching caused by histamine. This antihistamine is effective for the treatment of acute skin diseases such as urticaria.

However, there are also non-histamine dependent itching that are resistant to antihistamines. As non-histamine dependent itching, chronic itching such as atopic dermatitis, dry skin and itching caused by perspiration are known.
There are many receptors other than histamine receptors in sensory nerves that recognize itching, one of which is known as MrpprX1. This Mrgpr X1 is activated by chloroquine and bovine adrenal medullary peptide (8-22) (hereinafter, also referred to as "BAM (8-22)"). Further, it is described in Non-patent document 1 that itching is caused via sensory nerves such as C fibers when MrgprX1 is activated, and that the antihistamine is less effective against this itching. Further, it is described in Non-Patent Document 2 that mice lacking mouse MrgprA3, which corresponds to human MrgprX1, do not cause chronic itching caused by atopic dermatitis or dry skin.

  Therefore, inhibiting MrgprX1 activity is very important in preventing or ameliorating chronic itching. To date, 3-substituted-2- (diphenylmethyl) -1-azabicyclo [2.2.2] octane (3-substituted-2- (diphenylmethyl) -1-) was used to inhibit MrgprX1 activity and alleviate itching. The administration of compounds such as azabicyclo [2.2.2] octane) is described in US Pat.

International Publication No. 2010/065085

J. Neurosci., 2011, 31 (20), p. 7563-7567 Nat. Neurosci., 2013, 16 (2), p. 174-182

An object of the present invention is to provide an MrpprX1 activity inhibitor that inhibits MrpprX1 activity.
Another object of the present invention is to provide an inhibitor of calcium ion influx into cells via MrpprX1, which inhibits calcium ions from influx into cells via MrpprX1.
Another object of the present invention is to provide an itch preventing or ameliorating agent capable of preventing or improving skin itching at an early stage.

The present inventors diligently studied in view of the above problems. As a result, it has been found that the compounds represented by the following formulas (1) to (4) inhibit the activity of MrgprX1 and have an effect of suppressing the influx of calcium ions into cells via MrgprX1. Furthermore, it has been found that these compounds have the effect of preventing or improving skin itching.
The present invention has been completed based on these findings.

The present invention comprises a compound represented by the following formula (1), a compound represented by the following formula (2), a compound represented by the following formula (3), and a compound represented by the following formula (4) The present invention relates to a Mrppr X1 activity inhibitor comprising, as an active ingredient, at least one compound selected from the group.
The present invention also relates to a compound represented by the following formula (1), a compound represented by the following formula (2), a compound represented by the following formula (3), and a compound represented by the following formula (4) The present invention relates to an inhibitor of calcium ion influx into cells via Mrgpr X1, comprising as an active ingredient at least one compound selected from the group consisting of
Furthermore, the present invention relates to a compound represented by the following formula (1), a compound represented by the following formula (2), a compound represented by the following formula (3), and a compound represented by the following formula (4) The present invention relates to an itch preventing or improving agent comprising, as an active ingredient, at least one compound selected from the group consisting of

The MrpprX1 activity inhibitor of the present invention can inhibit MrgprX1 activity.
The inhibitor of calcium ion influx into cells via MrpprX1 of the present invention can inhibit calcium ions from influx into cells via MrpprX1.
Furthermore, the itch preventing or improving agent of the present invention can prevent or improve skin itching at an early stage.

7 is a graph showing the results of Test Example 3; FIG. 1 (a) is a graph showing a change in relative fluorescence intensity ratio when chloroquine is added to a cell expressing MrgprX1. FIG. 1 (b) is a graph showing the change in relative fluorescence intensity ratio when BAM (8-22) is added to a cell expressing MrgprX1. FIG. 1 (c) is a graph showing the change in relative fluorescence intensity ratio when histamine is added to cells in which H1R is expressed. FIG. 1 (d) is a graph showing the change in relative fluorescence intensity ratio when ATP is added to cells in which P2Y2 is expressed. 7 is a graph showing the results of Test Example 5; Fig.2 (a) is a graph which shows the MrpprX1 activity inhibitory effect of the compound represented by Formula (1)-(4). FIG.2 (b) is a graph which shows the H1R activity inhibitory effect of a compound represented by Formula (1)-(4). FIG.2 (c) is a graph which shows the P2Y2 activity inhibitory effect of the compound represented by Formula (1)-(4). 7 is a graph showing the results of Test Example 6; FIG. 3 (a) is a graph showing the change in the total score of itching when the compound of formula (1) is applied to the skin on which itching has been induced. FIG. 3 (b) is a graph showing a change in the total score of itching when Musk ketone is applied to the skin on which itching is caused.

As used herein, “MrgprX1” means Mas-related G-protein coupled receptor member X1 (Mas-related G-protein coupled receptor member X1). And this MrpprX1 is a protein encoded by the MRGPRX1 gene in humans.
MrgprX1 is a receptor specific for sensory neurons involved in itching and pain and is known to control noxious stimuli. Specifically, when an agonist such as chloroquine or BAM (8-22) binds to MrgprX1, MrgprX1 is activated. Activation of MrgprX1 stimulates the activation of the ion channel and the endoplasmic reticulum, which are involved in the control of intracellular calcium concentration. These series of processes induce an increase in intracellular calcium ion concentration in cells expressing MrgprX1, depolarization occurs, and pain and itch are caused by the firing of sensory nerves involved in nociception.
In the present specification, “in that calcium ions flow into cells via MrgprX1” refers to activation of MrgprX1 to stimulate the activation of ion channels and stimulation of the endoplasmic reticulum, resulting in an increase in intracellular calcium ion concentration. Refers to being triggered.

As used herein, "preventing" refers to preventing or delaying the onset of a disease or condition in an individual, or reducing the risk of developing an individual's disease or condition.
Moreover, in the present specification, “improvement” refers to amelioration of disease, symptom or condition, prevention or delay of deterioration of disease, symptom or condition, or reversal, prevention or delay of progression of disease, symptom or condition.
Furthermore, in the present specification, “non-therapeutic” is a concept that does not include medical practice, that is, treatment of the human body by treatment.

  The MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells via MrpprX1, and the itch-preventing or alleviating agent are compounds represented by the formula (1), represented by the formula (2) At least one compound selected from the group consisting of a compound, a compound represented by Formula (3), and a compound represented by Formula (4) is used as an active ingredient. In addition, at least one selected from the group consisting of a compound represented by the formula (1), a compound represented by the formula (2), a compound represented by the formula (3), and a compound represented by the formula (4) By applying a compound of a species, it is possible to inhibit MrgprX1 activity, to suppress the influx of calcium ions into cells through MrpprX1, and to prevent or improve itching.

The compounds used in the present invention may be commercially available products, or may be produced according to a conventional method. In the present invention, one of the above compounds may be used alone, or two or more may be used in combination. In addition, isomers such as geometric isomers, optical isomers, stereoisomers, etc. exist in the compound. Here, the compounds used in the present invention are not limited to the compounds represented by formulas (1) to (4), but also include isomers of these compounds. Furthermore, the compound used in the present invention may be any isomer or a mixture of isomers.
Each of the compounds is a highly safe compound conventionally used as a fragrance. Here, the common name of the said compound, an IUPAC name, etc. are shown below.

Compound Represented by Formula (1) Galaxolide: International Flavor and Fragrance
1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta (γ) -2-benzopyran (1,3,4,6,7,8- Hexahydro-4,6,6,7,8,8-hexamethylcyclopenta (γ) -2-benzopyran)

Compound represented by formula (2): Diacetyl (Diacetyl)
2,3-butanedione

The compound represented by the formula (3) : Suzalal: Takasago Fragrance Industry Co., Ltd. Silvial: Silvial
2-Methyl-3- [4- (2-methylpropyl) phenyl] propanal (2-methyl-3- [4- (2-methylpropyl) phenyl] propanal)

Compound represented by Formula (4) : Trans-2-decalal (Trans-2-decenal)
Deck-2-enal (Dec-2-enal)

As demonstrated in the below-mentioned Example, the compound represented by each of Formula (1)-(4) inhibits MrgprX1 activity, and suppresses the influx of the calcium ion in the cell via MrgprX1. Furthermore, the compounds represented by each of the formulas (1) to (4) having the action of inhibiting MrpprX1 activity suppress and alleviate the itching of the skin caused by activating MrpprX1 in a short time. Therefore, the compounds represented by each of the formulas (1) to (4) are useful for the inhibition of MrgprX1 activity, the suppression of the influx of calcium ions through MrgprX1 and the prevention or amelioration of itch.
In addition, as demonstrated in the below-mentioned Example, the compound represented by each Formula (1)-(4) acts on MrgprX1, and inhibits MrgprX1 activity. However, the compounds represented by each of the formulas (1) to (4) do not act on histamine receptor H1R or ATP receptor which is the same G protein as MrgprX1. Here, the histamine receptor H1R or ATP receptor is a receptor that induces itching due to inflammation or cell injury or the like in which the production of histamine or ATP is induced. MrgprX1 contrast, a receptor that induces pruritus by atopic dermatitis. Therefore, the compounds represented by each of the formulas (1) to (4) which do not act on histamine receptor H1R or ATP receptor but inhibit MrgprX1 activity on MrgprX1 are particularly nonhistamines which are not effective against antihistamines dependent chronic itching (atopic dermatitis, senile pruritus, contact dermatitis, prurigo, pruritus accompanying sweating, itching in sensitive areas (genital)) is particularly useful for prevention or amelioration of such.

  The form of the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells through MrpprX1, and the agent for preventing or improving pruritus can be appropriately selected. For example, the active ingredient alone may be used as the MrpprX1 activity inhibitor of the present invention, an inhibitor of calcium ion influx into cells via MrpprX1, and a pruritus-preventing or improving agent. Alternatively, a pharmaceutical composition of the MrpprX1 activity inhibitor of the present invention comprising the active ingredient and a pharmaceutically acceptable carrier, an inhibitor of calcium ion influx into cells through MrpprX1, and an agent for the prevention or improvement of itch You may use it as a thing. Alternatively, the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells through MrpprX1, and the agent for preventing or improving itch may be used as a cosmetic composition, or may be contained in them. .

  When preparing a pharmaceutical composition, it is usually prepared as a preparation containing the active ingredient and preferably a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier generally refers to an inert, non-toxic, solid, liquid, bulking agent, diluent or encapsulating material which does not react with the active ingredient, such as water And solvents such as ethanol, polyols (eg, propylene glycol, butylene glycol, glycerin, polyethylene glycol and the like), suitable mixtures thereof, solvents or dispersion media such as vegetable oils, and the like.

  The pharmaceutical composition may be orally, parenterally, for example, in the oral cavity, in the skin, subcutaneously, in the mucous membrane, intravenously, in artery, intramuscularly, intraperitoneally, vaginally, in the lung, Administered in the brain, in the eye, or in the nasal cavity. The oral administration preparations include tablets, granules, fine granules, powders, capsules, chewables, pellets, syrups, solutions, suspensions and inhalants. For parenteral administration, suppositories, holding enemas, drips, eye drops, nasal drops, pessaries, injections, mouthwashes, as well as ointments, creams, lotions, gels, controlled release patches And external preparations for skin such as patches. The pharmaceutical composition may be administered parenterally in the form of a sustained release subcutaneous implant, or in the form of a targeted delivery system (eg, monoclonal antibody, vector delivery, ion injection, polymer matrix, liposomes and microspheres) .

  The pharmaceutical composition may further contain conventional additives in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, coloring agents, flavoring agents and the like, which can be used if necessary. In order to make a sustained release so as to be able to act for a long time, it can be coated with a known retarder or the like. As the excipient, for example, sodium carboxymethylcellulose, agar, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, magnesium aluminometasilicate, calcium hydrogen phosphate etc. It can be used. Examples of binders include gum arabic, sodium alginate, ethylcellulose, sodium caseinate, sodium carboxymethylcellulose, agar, purified water, gelatin, starch, tragacanth, lactose and the like. As the disintegrant, for example, carboxymethylcellulose, carboxymethylcellulose sodium, carboxymethylcellulose calcium, crystalline cellulose, starch, hydroxypropyl starch and the like can be mentioned. As the lubricant, for example, stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, waxes and the like can be mentioned. Examples of the antioxidant include tocopherol, gallic acid ester, dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), ascorbic acid and the like. Other additives and drugs as required, such as antacid (sodium hydrogen carbonate, magnesium carbonate, precipitated calcium carbonate, synthetic hydrotalcite, etc.), gastric mucosa protective agent (synthetic aluminum silicate, sucralfate, sodium copper chlorophyllin etc.) ) May be added.

  When the cosmetic composition is prepared, the form can be appropriately selected, and solutions, emulsions, powders, water-oil two-layer systems, water-oil-powder three-layer systems, gels, tablets such as solids, aerosols, It can be in any form such as mist, capsule and sheet. In addition, the product form of the cosmetic composition is also optional, for example, cleanser, makeup remover, lotion, cosmetic solution, pack, milk, skin care cosmetic such as cream and sunscreen, foundation, makeup base, lipstick, eye Makeup cosmetics such as shadows, eyeliners, mascara, eyebrows, blushers and nail enamels, hair shampoos, hair rinses, hair conditioners, hair cosmetics such as hair dyes and hair restorers, soaps, body soaps, deodorant cosmetics and bath preparations And body cosmetics, oral cosmetics such as dentifrices and mouthwashes, and aromatic cosmetics such as perfumes. Also, the cosmetic may belong to either cosmetic or quasi-drug under the Japanese Pharmaceutical Affairs Law.

The cosmetic composition includes other components commonly used in cosmetics, quasi-drugs and pharmaceuticals, such as powder components, liquid fats and oils, solid fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, anions Surfactant, Cationic surfactant, Amphoteric surfactant, Non-ionic surfactant, Moisturizer, Water-soluble polymer, Thickener, Coating agent, UV absorber, Sequestering agent, Lower alcohol, Polyhydric alcohol Sugar, amino acids, organic amines, polymer emulsions, pH adjusters, skin nutrients, vitamins, antioxidants, antioxidant aids, perfumes, water, etc., if necessary, and manufactured by a conventional method it can.
Examples of components that can be added to other cosmetic compositions include preservatives (eg, ethyl paraben, butyl paraben, etc.), anti-inflammatory agents (eg, glycyrrhizinic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin etc. ), Whitening agents (eg, ascorbic acid and derivatives thereof, placenta extract, yukinoshita extract, arbutin etc.), various extracts (eg, oubac, ouren, sikon, peony, cerebrum, birch, sage, loquat, carrot, aloe) , Zebra oasis, iris, grapes, yokinin, oats, lilies, saffron, sage, sage, oonilis, ononis, garlic, chili pepper, chili, seaweed etc., activators (eg royal jelly, photosensitizer, cholesterol derivative etc) , Blood circulation enhancer For example, nonyl acid warylamide, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, zingerone, cantalistinqui, ictamol, tannic acid, α-borneol, nicotinic acid tocopherol, inositol hexanicotinate, cichrandelate, cinnarizine , Trazoline, acetylcholine, verapamil, cephalantin, γ-oryzanol etc., antiseborrhoeic agent (eg sulfur, thianthol etc.), anti-inflammatory agent (eg tranexamic acid, thiotaurine, hypotaurine etc) and bactericidal agent (eg triclosan, And cetyl pyridinium chloride, thymols, benzalkonium chloride and the like.

The pharmaceutical composition and the cosmetic composition can be applied in the form of an oral composition, an external composition, an internal use composition, etc., and it is preferable to apply in the form of a skin external composition.
When used in the form of a composition for external application to the skin, in addition to the above-mentioned active ingredients, components used for the composition for external application to the skin, such as surfactants, oily substances, polymer compounds, preservatives, various medicinal components, powders An ultraviolet absorber, a pigment, a flavor, an emulsion stabilizer, a pH adjuster, etc. can be suitably blended.

  The MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells through MrpprX1, and the agent for preventing or improving pruritus are used by being added or formulated to foods, beverages, feeds, pet foods Can. Alternatively, it was displayed as a concept for treating, preventing or ameliorating a disease or condition that may be treated, prevented or ameliorated by inhibition of MrgprX1 activity and / or suppression of calcium ion influx into cells via MrgprX1. It can be used by being added to or mixed with food and drink, that is, health food, functional food, food for the sick, food for specified health and the like. The above-mentioned health food, functional food, food for sick persons, food for specified health use, specifically, various kinds of fine granules, tablets, granules, powders, capsules, syrups, solutions, liquid foods, etc. It can be used as a pharmaceutical form. The food in the form of a preparation can be produced in the same manner as a pharmaceutical preparation, and the active ingredient and a food-acceptable carrier such as a suitable excipient (eg starch, processed starch, lactose, glucose, water etc.), etc. And can be manufactured using conventional means. Furthermore, liquid food compositions such as soups, juices, milk drinks, tea drinks, coffee drinks, cocoa drinks, jelly drinks, sports drinks, diet drinks, semi-solid food compositions such as puddings and yogurts, breads, For any noodles, cookies, chocolates, candies, gums, confections such as rice crackers, spreads such as sprinkles, butter, jams, etc., the MrgprX1 activity inhibitor of the present invention, calcium ion into cells via MrpprX1 A food composition can be produced by adding or blending inflow inhibitors, or itch-preventing or improving agents.

The content of the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells via MrpprX1, and the content of the active ingredient in the agent for preventing or improving itch can be determined appropriately.
For example, the content of the active ingredient is 0.001% by mass or more in the total amount of the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells through MrpprX1, and the itch preventing or improving agent. 0.05 mass% or more is more preferable, 0.1 mass% or more is more preferable, 10 mass% or less is preferable, 1 mass% or less is more preferable, 0.5 mass% or less is more preferable, 0.001 -10 mass% is preferable, 0.05-1 mass% is more preferable, 0.1-0.5 mass% is further more preferable.

  The subject of administration or intake of the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells via MrpprX1, and the agent for the prevention or improvement of itch is preferably a warm-blooded vertebrate, more preferably a mammal It is an animal. In the present specification, mammals include, for example, humans and non-human mammals such as monkeys, mice, rats, rabbits, dogs, cats, cattle, horses, and pigs. The MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells via MrpprX1, and the agent for preventing or improving pruritus are suitable for human administration.

The above-mentioned compounds used in the present invention, the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells through MrpprX1 and the itch preventing or improving agent are itchy skin, preferably non-histamine on skin It can be preferably applied to a subject who desires to suppress dependence on chronic itching. The compound used in the present invention, the MrpprX1 activity inhibitor of the present invention, an inhibitor of calcium ion influx into cells through MrpprX1, and an itch preventing or improving agent are itchy skin, preferably skin itch conditions chronic itching non histamine independent, is raised, preferably to sweating conditions, in preferred applications. Furthermore, the compound used in the present invention, the MrpprX1 activity inhibitor of the present invention, the inhibitor of calcium ion influx into cells via MrpprX1, and the itch-preventing or alleviating agent are preferably applied to the skin , skin itching due to sweating is occurring, or is more preferable to apply to the skin that tends to occur is itching associated with sweating.

In the method for inhibiting MrpprX1 activity of the present invention, a method for inhibiting calcium ion influx into cells via MrpprX1, and a method for preventing or improving itch, the effective amount of the active ingredient to be applied by administration or ingestion is The condition, body weight, sex, age, activity of material, administration or intake route, administration or intake schedule, formulation form or other factors can be appropriately determined. For example, the effective amount of the active ingredient is preferably 0.0015 mg or more, more preferably 0.075 mg or more, preferably 22.5 mg or less, more preferably 2.4 mg or less, per kg of body weight per day. Is 0.0015 to 22.5 mg, more preferably 0.075 to 2.4 mg. The active ingredient may be taken / administered once to several times a day, or at any time and interval.
In the present invention, the dose of the active ingredient is as described in J. Neurosci., 2011, 31 (20), p. It can be determined as appropriate according to a conventional method such as the method described in 7563-7567. In addition, administration or intake of the active ingredient may be systemic administration or intake, or may be local administration or intake.

  With regard to the above-described embodiment, the present invention further discloses the following MrgprX1 activity inhibitor, an inhibitor of calcium ion influx into cells through MrpprX1, an agent for preventing or improving pruritus, uses, and methods.

<1> A Compound Represented by Formula (1), a Compound Represented by Formula (2), a Compound Represented by Formula (3), and a Compound Represented by Formula (4) An MrpprX1 activity inhibitor, an inhibitor of calcium ion influx into cells through MrpprX1, or an agent for preventing or improving pruritus, which comprises one compound as an active ingredient.

<2> The Mrppr X1 activity inhibitor according to <1>, wherein the MrpprX1 is human-derived MrpprX1, or an inhibitor of the influx of calcium ions into cells via MrpprX1.
The itch preventing or ameliorating agent according to <1>, which prevents or improves itching by inhibiting <3> MrgprX1 activity.
<4> the itching, chronic itching non histamine-dependent, preferably atopic dermatitis, senile pruritus, contact dermatitis, prurigo, pruritus accompanying sweating or itching in sensitive areas, and more preferably itching associated with sweating, and the <1> or <3> itching preventive or ameliorating agent according to claim.
The content of the said active ingredient is 0.001 mass% or more in the total amount of <5> said MrpprX1 activity inhibitor, the inhibitor of the inflow of the calcium ion to the inside of a cell through MrpprX1, or itch prevention or improvement agent, Preferably, it is 0.05% by mass or more, more preferably 0.1% by mass or more, and 10% by mass or less, preferably 1% by mass or less, more preferably 0.5% by mass or less. The MrpprX1 activity inhibitor according to any one of <1> to <4>, an inhibitor of calcium ion influx into cells via MrpprX1, or an itch-preventing or ameliorating agent.

The compound represented by Formula (1) as a <6> MrgprX1 activity inhibitor, the inhibitor of the inflow of the calcium ion to the inside of a cell through MrpprX1, or itch, or it is represented by Formula (2) by Formula (2) Use of at least one compound selected from the group consisting of compounds represented by formula (3), compounds represented by formula (3), and compounds represented by formula (4).
<7> MrpprX1 activity inhibitor, inhibitor of calcium ion influx into cells through MrpprX1, or a compound represented by the formula (1) for the production of an itch preventing or improving agent, the formula (2) Use of at least one compound selected from the group consisting of a compound represented by :, a compound represented by Formula (3), and a compound represented by Formula (4).
<8> Compound Represented by Formula (1), Compound Represented by Formula (2), Compound Represented by Formula (3), and Compound Represented by Formula (4) A method of using one type of compound as an MrpprX1 activity inhibitor, an inhibitor of intracellular calcium ion influx through MrpprX1, or an agent for preventing or improving pruritus.
<9> Compound Represented by Formula (1), Compound Represented by Formula (2), Compound Represented by Formula (3), and Compound Represented by Formula (4) An MRppr X1 activity inhibition method, a method of suppressing calcium ion influx into cells via Mrppr X1, or a method of preventing or improving pruritus, which comprises applying one compound.
<10> The use or the method according to any one of the above <6> to <9>, wherein the MrpprX1 is human-derived MrgprX1.
The use or method of any one of the above <6> to <10>, which prevents or ameliorates itching by inhibiting <11> Mrgpr X1 activity.
<12> the itching, chronic itching non histamine-dependent, preferably atopic dermatitis, senile pruritus, contact dermatitis, prurigo, pruritus accompanying sweating or itching in sensitive areas, and more preferably itching associated with sweating, and the <6> - use or method according to any one of <11>.
<13> The compound according to any one of <8> to <12>, which is applied to human desiring suppression of skin itching, preferably non-histamine dependent chronic itching of the skin. Method.
<14> itchy skin, preferably itching non histamine-dependent skin conditions were elicited, preferably sweating conditions, in applying, in any one of the above <8> ~ <13> Method described.
<15> applying the compounds skin, preferably skin itching accompanying sweating occurs, or itching prone skin due to sweating, in the <8> ~ any one of <14> The method described in.
<16> The content of the compound is preferably 0.001% by mass or more in the total amount of the MrpprX1 activity inhibitor, the inhibitor of calcium ion influx into cells through MrpprX1, or itch-preventing or alleviating agents. Is 0.05% by mass or more, more preferably 0.1% by mass or more, and 10% by mass or less, preferably 1% by mass or less, more preferably 0.5% by mass or less. -The use or method of any one of <15>.

<17> A method of inhibiting MrpprX1 activity, a method of inhibiting calcium ion influx into cells via MrpprX1, or a compound represented by formula (1), used for preventing or improving itch, At least one compound selected from the group consisting of a compound represented by the present invention, a compound represented by the formula (3), and a compound represented by the formula (4).
<18> MrpprX1 activity inhibitor, inhibitor of calcium ion influx into cells through MrpprX1, or a compound represented by the formula (1) for the preparation of a drug for preventing or improving itch, formula (2) Use of at least one compound selected from the group consisting of a compound represented by :, a compound represented by Formula (3), and a compound represented by Formula (4).
<19> A compound represented by the formula (1), which is used for non-therapeutic treatment method of inhibition of MrgprX1 activity, suppression of calcium ion influx into cells through MrpprX1, or prevention or amelioration of itch, Use of at least one compound selected from the group consisting of a compound represented by Formula (2), a compound represented by Formula (3), and a compound represented by Formula (4).
<20> The compound or use according to any one of the above <17> to <19>, wherein the MrpprX1 is human-derived MrgprX1.
The compound or use of any one of the above <17> to <20>, which prevents or ameliorates itching by inhibiting <21> Mrgpr X1 activity.
<22> the itch, non-histamine-dependent itching, preferably atopic dermatitis, senile pruritus, contact dermatitis, prurigo, pruritus accompanying sweating or itching in sensitive areas, and more preferably in perspiration The compound or use according to any one of the above <17> to <21>, which is itching along with it.
<23> The compound according to any one of <17> to <22>, wherein the compound is applied to humans in which it is desired to suppress skin itching, preferably non-histamine dependent chronic itching of the skin. Compound or use.
<24> itchy skin the compound, preferably under conditions non-histamine-dependent chronic itchy skin, is raised, preferably applied in sweating conditions, the <17> - <23> Or a compound according to any one of the preceding claims.
<25> applying the compounds skin, preferably skin itching accompanying sweating occurs, or itching prone skin due to sweating, in the <17> - any one of <24> The compound or use as described in.
<26> The compound or use according to any one of the above <17> to <25>, wherein the compound is applied in the form of a pharmaceutical composition or a cosmetic composition.
<27> The compound or use according to the above <26>, wherein the compound is applied in the form of a composition for external application to the skin.
The compound or use of any one of the said <17>-<25> which applies the <28> above-mentioned compound in the form of foodstuffs, a drink, or feed.
The content of the <29> compound is 0.001% by mass or more, preferably 0.05% by mass or more, more preferably 0.1% by mass or more, and 10% by mass or less, preferably 1% by mass or less The use according to any one of <17> to <28>, which is more preferably 0.5% by mass or less.

<30> At least one selected from the group consisting of a compound represented by Formula (1), a compound represented by Formula (2), a compound represented by Formula (3), and a compound represented by Formula (4) A nontherapeutic method for inhibiting MrpprX1 activity, a nontherapeutic method for inhibiting the influx of calcium ions into cells via MrpprX1, or a nontherapeutic method for preventing or ameliorating itch, wherein an effective amount of one compound is applied.
<31> The method according to the above <30>, wherein the MrpprX1 is human-derived MrgprX1.
The method according to <30> or <31>, wherein itch is prevented or ameliorated by inhibiting <32> MrgprX1 activity.
<33> the itching, chronic itching non histamine-dependent, preferably atopic dermatitis, senile pruritus, contact dermatitis, prurigo, pruritus accompanying sweating or itching in sensitive areas, and more preferably itching associated with sweating, and the <30> - the method according to any one of <32>.
<34> The compound according to any one of <30> to <33>, which is applied to human desiring suppression of skin itching, preferably non-histamine dependent chronic itching of skin. Method.
<35> itchy skin, preferably a non-histamine-dependent itchy skin, conditions were elicited, preferably sweating conditions, in applying, the <30> - to any one of <34> Method described.
<36> the skin of said compounds, is preferably applied skin itching accompanying sweating occurs, or itching prone skin due to sweating, in the <30> - any one of <35> The method described in.
<37> The effective amount of the compound is 0.0015 mg or more, preferably 0.075 mg or more, and 22.5 mg or less, preferably 2.4 mg or less, per 1 kg of body weight per day. 30. The method according to any one of <36>.

  EXAMPLES The present invention will be described in more detail based on examples given below, but the invention is not meant to be limited by these.

(Test Example 1) Cloning of Receptor Gene Using cDNA obtained by reverse transcribing total RNA of human neuroblastoma SK-N-MC cells as a template, PrimeSTAR GXL DNA Polymerase (trade name, manufactured by TaKaRa) is used. The MRGPRX1 gene, the H1R gene and the P2Y2 gene were amplified by polymerase chain reaction (PCR).
The PCR conditions were: 98 ° C. 2 minutes → [98 ° C. 15 seconds, 68 ° C. 1.5 minutes] × 30 cycles → 68 ° C. 2 minutes. The base sequences of various primers used in PCR are shown in Table 1 below.

The obtained PCR product was purified using High Pure PCR Product Purification Kit (trade name, manufactured by Roche). Then, using the pcDNA3.1 Directional TOPO Expression Kit (trade name, manufactured by Life technologies), the purified PCR product is inserted into an expression vector (trade name: pcDNA3.1D / V5His-TOPO, manufactured by Life technologies). Then, a plasmid for receptor gene expression was constructed. The sequence analysis of the insert of the receptor gene was consigned to Takara Bio Inc., and it was confirmed that the human MRGPRX1 gene, H1R gene and P2Y2 gene were correctly inserted into the expression vector.

Test Example 2 Introduction of Receptor Gene into HEK 293 Cells and Strong Expression of Receptor Gene The E. coli JM109 strain was transformed using each of the receptor gene expression plasmids constructed in Test Example 1. Then, using NucleBond Xtra Midi EF (trade name, manufactured by TaKaRa), a large amount of a plasmid for receptor gene expression was extracted and purified.

Human embryonic kidney cells HEK293 were cultured in T-75 cell culture flasks. Then, 8 μg of the plasmid into which each receptor gene was inserted was introduced into 70-80% confluent HEK 293 using a transfection reagent (trade name: TransIT-293, manufactured by Mirus) and cultured for 24 hours.
The adhered cells were detached from the culture flask using an adherent cell dispersion reagent (trade name: Detachin, manufactured by Genlantis). Then, the detached cells were seeded on a 96 well Optical Bottom Plate (trade name, manufactured by Nunc) at 1.5 × 10 ⁴ cells / well and cultured for further 24 hours.
As a control, a plasmid into which lacZ has been inserted (trade name: pcDNA3.1D / V5-His / lacZ, manufactured by Life technologies) is introduced into human embryonic kidney cells HEK293, and cells are prepared by the same method as described above. did.

(Test Example 3) Measurement of Receptor Activity As an indicator of receptor activation, the influx of calcium ions into cells in which each receptor was strongly expressed was measured using Calcium Kit II-fluo-4 (trade name, DOJINDO). Measurement). The measurement operation was performed according to the protocol of the kit.
Loading buffer containing a calcium fluorescent indicator (trade name: fluo-4-AM) was added to the cells prepared in Test Example 2 and incubated at 37 ° C. for 1 hour. Thereafter, the fluorescence intensity was measured by FDSS (manufactured by Hamamatsu Photonics K.K.) at 37.degree.
Thirty seconds after the start of measurement, an agonist of the receptor shown in Table 2 was added to cells expressing any of the MrpprX1, H1R and P2Y2 receptors. Thereafter, the fluorescence intensity was measured every two seconds for 300 seconds. The relative fluorescence intensity ratio to the fluorescence intensity before adding the agonist was calculated assuming that the fluorescence intensity before adding the agonist was 1.
The results are shown in FIG. FIG. 1 (a) is a graph showing the change in relative fluorescence intensity ratio when chloroquine is added as a receptor agonist to cells in which MrpprX1 is expressed. FIG. 1 (b) is a graph showing the change in relative fluorescence intensity ratio when BAM (8-22) is added as a receptor agonist to cells in which MrgprX1 is expressed. FIG. 1 (c) is a graph showing the change in relative fluorescence intensity ratio when histamine is added as a receptor agonist to cells in which H1R is expressed. FIG. 1 (d) is a graph showing the change in relative fluorescence intensity ratio when ATP is added as a receptor agonist to cells in which P2Y2 is expressed.

The activity of human-derived MrgprX1, H1R and P2Y2 was measured based on the change in the amount of calcium flowing into cells. As a result, as shown in FIGS. 1 (a) to (d), it was confirmed that the addition of an agonist for each receptor increased the fluorescence intensity depending on the concentration of the agonist to be added. That is, each receptor was confirmed to be activated by adding an agonist.
When a plasmid into which lacZ was inserted (trade name: pcDNA3.1D / V5-His / lacZ) was introduced as a control, no change in fluorescence intensity was detected.

(Test Example 4) Search for MrpprX1 Activity Inhibitor In the above Test Example 3, when BAM (8-22) is added as an agonist of MrpprX1, the final concentration of BAM (8-22) at which MrgprX1 activity becomes around EC ₅₀ is It was 0.5 μM.
Therefore, Loading buffer containing a calcium fluorescent indicator (trade name: fluo-4-AM) and BAM (8-22) to a final concentration of 0.5 μM were added to the cells expressing MrgprX1. . Various materials were further added to this, and the relative fluorescence intensity ratio was measured in the same manner as in Test Example 3. Then, based on the relative fluorescence intensity ratio measured, a search was made for an MrpprX1 activity inhibitor.

As a result, the compound represented by Formula (1)-(4) was discovered as a raw material which inhibits 80% or more of MrpprX1 activity.
In addition, the compound represented by Formula (1)-(4) is conventionally used as a fragrance | flavor. On the other hand, Musk ketone (1- (4-tert-butyl-2,6-dimethyl-3,5-dinitrophenyl), which has been conventionally used as a flavor as in the compounds represented by the formulas (1) to (4) The inhibition rate of Mrppr X1 activity of Ethanone was 11.8%.

Test Example 5 Inhibition Test of Receptor Activity
A loading buffer containing a calcium fluorescent indicator (trade name: fluo-4-AM) was added to the cells in which MrgprX1 is expressed, the cells in which H1R is expressed, and the cells in which P2Y2 is expressed. Here, a mixed solution of BAM (8-22), histamine or ATP of each receptor agonist and each compound represented by formulas (1) to (4) was added. In addition, the addition amount of the agonist of each receptor was suitably set so that the activity of each receptor might be EC ₅₀ vicinity from the result of Experiment 4. Moreover, the addition amount of the compound represented by Formula (1)-(4) was set so that final concentration might be 0 to 0.01%.
Then, after the addition of the mixed solution, the fluorescence intensity was measured for 300 seconds, and the maximum value of the relative fluorescence intensity ratio was measured. Furthermore, the value of the maximum relative fluorescence intensity ratio at a compound concentration of 0% was set to 1.0, and each receptor activity was calculated. The results are shown in FIG.

As shown in FIG. 2, the MrpprX1 activity was inhibited depending on the concentrations of these compounds by adding the compounds represented by formulas (1) to (4). On the other hand, the compounds represented by the formulas (1) to (4) hardly acted on H1R and P2Y2, and no action to inhibit H1R activity and P2Y2 activity was observed.
From the above results, it was confirmed that the compounds represented by the formulas (1) to (4) specifically act on MrpprX1 among G protein-coupled receptors and inhibit MrgprX1 activity.

(Test Example 6) Itch induction test by application of BAM (8-22), and prevention or improvement test of itch
J. Neurosci., 2011, 31 (20), p. With reference to the method described in 7563-7567 (Non-Patent Document 1), itching test by application of BAM (8-22) and prevention or improvement test of itching were performed.

  After applying 10 μL of ethanol to human skin (the upper arm extension side) and leaving it to stand for 1 minute, prick lancet pierces the same site three times at 1 mm intervals, and after 30 seconds, 5 mM BAM (8-22) HEPES solution 5 μL was applied. Every 30 seconds from the application of BAM (8-22), it was evaluated for 5 minutes for a stagnation score of 0 to 10 (0: not considered, 10: the worst so far). Then, the sum of the stagnation scores evaluated every 30 seconds was calculated as a swallow total score.

On the other hand, 10 μL of an ethanol solution (concentration: 0.1%) of the compound represented by the formula (1) is applied before applying the above-mentioned BAM (8-22), and after standing for 1 minute, the above-mentioned BAM (8-22) was applied as per. Then, the same operation as described above was performed, the stagnation score of 0 to 10 was individually evaluated, and the stagnation total score was calculated.
These results are shown in FIG. 3 (a).

  The itch score was calculated in the same manner as described above, except that Musk ketone was used instead of the compound represented by Formula (1). The result is shown in FIG. 3 (b).

As shown in FIG. 3, itching was induced by applying BAM (8-22) to the skin. On the other hand, as shown in FIG. 3A, by applying the compound represented by the formula (1), the total score for stagnation decreased significantly in a short time. This indicates that the itching induced by the application of BAM (8-22) is relieved in a short time by applying the compound represented by the formula (1).
On the other hand, as shown in FIG. 3 (b), even if application of Musk ketone having no action to inhibit Mrgpr X1 activity in place of the compound represented by formula (1), the itch score is remarkable. No decrease was observed, and the itching induced by the application of BAM (8-22) was not alleviated.

(Test Example 7) itching of preventing or of compositions shown in improving test Table 3 Body lotion caused by sweating (Galaxolide (G) and Placebo (P)) was prepared,
The bottle was provided in a mist spray bottle (discharge volume per push: 0.1 mL).

Tests for preventing or improving itching were conducted in winter according to the schedule shown below for one woman who felt itching due to perspiration on the trunk more than once a day in summer.

From the weekend immediately prior to use a body lotion of the test product, stop the care of to the trunk, which is doing usually, Placebo (P) Body Lotion the torso (neck, chest, waist) to 5 days (month ~ Friday) used.
And next week (Mon-Fri), Galaxolide (G) body lotion was used as well.
The body lotion was used three times a day before work in the morning, during work in the daytime, and after work in the evening. The amount of body lotion used per day was limited to 2.0 mL each. Also, the prevention or amelioration test for itching was a double-blind test , the test item was encoded at the bottling stage, and subjects and data analysts were not notified of the contents of the test item .

<Itch evaluation>
During the period you are using a body lotion of the test product, it was recorded every night of the degree of itching (subjective evaluation). The "average degree of itch felt in one day", itching strength (10cmVAS, left end (0): itching do not feel anything the Mi, right edge (10): very strong feel itching) and, itching score (0: itching None, 1: slight itching, 2: weak itching, 3: moderate itching, 4: strong itching, 5: very strong itching . The results are shown in Table 5 .

As shown in Table 5 , as a result of analyzing the average value of 10 cm VAS of sweating itching and itching score for 5 days, the numerical values of both 10 cm VAS and itching score decreased when using Galaxolide as compared with using Placebo. . Furthermore, when using Placebo, "there was a sense of incongruity in the neck and accompanied by itching", when using Galaxolide, "the feeling of the neck was felt in the evening was relieved in the evening, and the itching was not bothersome Response was received. In other words, it was felt that Galaxoldie was effective in relieving itching.

Degree of degree of sweating
During the period of using the body lotion of the test product, scores of degree of perspiration (subjective evaluation) were recorded every night (0: no sweat, 1: slightly sweated, 2: slightly sweated, 3: moderate sweating I did, 4: I sweated strongly, 5: I sweated very strongly).
As a result, the average score of the degree of sweating felt during the test period was 1.4 in the week using Placebo and 1.6 in the week using Galaxolide, and the amount of sweating tends to be larger in the week using Galaxolide. The That is, when using Galaxolide, the itching caused by sweating was alleviated despite the large amount of sweating.

As mentioned above, itchy skin was relieved in a short time by applying the compound which has the effect | action which inhibits MrgprX1 activity. This indicates that the compound having the activity of inhibiting MrpprX1 activity is effective for preventing or ameliorating skin itching. Furthermore, it has been shown that the compound having the activity of inhibiting MrpprX1 activity has an immediate action on itching, which is a reduction of itching in a short time.
Furthermore, the results of the above embodiment has an effect of inhibiting the MrgprX1 activity, the compounds defined in the present invention, for non-histamine-dependent chronic itching such as pruritus due to sweating, preventing or ameliorating It shows that it has an effect.

Claims (4)

A compound represented by formula (1),
A compound represented by formula (2),
An Mrppr X1 activity inhibitor comprising, as an active ingredient, at least one compound selected from the group consisting of a compound represented by the formula (3) and a compound represented by the formula (4).
A compound represented by formula (1),
A compound represented by formula (2),
Inflow of calcium ion into cells via Mrgpr X1 comprising as an active ingredient at least one compound selected from the group consisting of a compound represented by the formula (3) and a compound represented by the formula (4) Inhibitor.
A compound represented by formula (1),
A compound represented by formula (2),
An itch preventing or improving agent comprising, as an active ingredient, at least one compound selected from the group consisting of a compound represented by the formula (3) and a compound represented by the formula (4).
The itching prevention or remedy according to claim 3, wherein the itching is itching associated with sweating.