Prophylactic or ameliorating agent for itching

Abstract

The present invention provides a novel raw material useful for preventing or ameliorating itching by inhibiting a C3a receptor signaling pathway. The present invention is a C3a receptor antagonist containing telukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

Description

Technical Field

[0001] The present invention relates to a C3a receptor antagonist, an agent for preventing or improving itching, and an agent for preventing or improving pruritic skin diseases. Background Art

[0002] Itching is a condition that appears not only in skin diseases such as atopic dermatitis but also in visceral diseases such as renal failure. Itching can also be caused by skin dryness, sun exposure, friction between the skin and clothing, etc. Scratching due to itching physically damages the skin and further worsens the symptoms. Therefore, solving itching helps prevent or improve skin diseases. For example, it has been reported that in mice whose hind limb nails were cut to inhibit physical damage to the skin caused by scratching, skin symptoms caused by atopic dermatitis were prevented and improved (Non-Patent Document 1).

[0003] The sensation of itching in peripheral tissues such as the skin is transmitted to the brain through afferent sensory nerves that connect the periphery to the posterior horn of the spinal cord. The cell bodies of the afferent sensory nerves are present in the dorsal root ganglion (DRG), and nerve fibers extend from the cell bodies to the peripheral tissues and the posterior horn of the spinal cord. The afferent sensory nerves have the function of receiving sensations in the skin and transmitting them to the second-order neurons in the posterior horn of the spinal cord.

[0004] Pruritogenic substances cause itching by binding to corresponding receptors. As pruritogenic substances, histamine, serotonin, chloroquine, etc. have been reported, and as itching-enhancing substances (sensitizing substances), Th2 cytokines such as IL-4 or IL-13 have been reported. Among them, the representative chemical pruritogenic substance is mainly histamine secreted by mast cells. Recently, it has been considered that histamine is basically only related to some acute itching, and its involvement in most diseases accompanied by chronic itching is not significant (Non-Patent Document 2). Therefore, currently, antihistamines (H1 receptor antagonists) are mostly used as drugs to inhibit itching, but the itching for which sufficient therapeutic effects can be obtained by antihistamines is extremely limited, and most itching conditions are intractable itching that is not easily relieved by antihistamines. The antihistamines mentioned here refer to H1 receptor antagonists such as diphenhydramine. For example, it has been reported that the effects of treating atopic dermatitis and xeroderma with antihistamines are not sufficient (Non-Patent Documents 3 to 4), and itching accompanying most skin diseases such as atopic dermatitis and xeroderma, and furthermore, internal medicine diseases such as renal failure is called intractable itching. The mechanisms of most intractable itching have not been elucidated, and it is necessary to elucidate the mechanisms and develop new target molecules.

[0005] On the other hand, it has been reported that tipelukast has leukotriene receptor antagonistic action, phosphodiesterase (PDE, mainly -3 and 4) inhibitory action, 5-lipoxygenase inhibitory action, etc. Tipelukast inhibits the biosynthesis of leukotrienes via the 5-lipoxygenase (5-LO) pathway through these multiple mechanisms, thereby exhibiting an anti-inflammatory effect or a fibrosis preventive effect. In addition, an effect of reducing neutral fat in the blood is observed (Patent Documents 1 and 2).

[0006] (Patent Document 1) Japanese Patent Application Laid-Open No. 2007-524624

[0007] (Patent Document 2) Japanese Patent Application Laid-Open No. 2016-525546

[0008] (Non-Patent Document 1) Hashimoto Y et al. Life Sciences. 2004 Dec 31; 76(7):783-94

[0009] (Non-Patent Document 2) Ikoma A et al. Nature Reviews Neuroscience. 2006 Jul; 7(7):535-47

[0010] (Non-Patent Document 3) J Am Acad Dermatol. 2014 Jul; 71(1):116-132

[0011] (Non-Patent Document 4) Future Oncol. 2018 Oct; 14(24):2531-2541

[0012] (Non-Patent Document 5) Lewis JE et al. Front Endocrinol (Lausanne). 2015 Feb 2; 6:3

[0013] (Non-Patent Document 6) Cero C et al Structure. 2014 Dec 2; 22(12):1744-1753 SUMMARY OF THE INVENTION

[0014] The present invention relates to the following 1) to 3).

[0015] 1) A C3a receptor antagonist, which contains tipelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

[0016] 2) A preventive or ameliorating agent for itching, which contains tipelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

[0017] 3) A prophylactic or ameliorating agent for pruritic skin diseases, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Showing the C3a receptor antagonistic effect of montelukast.

[0019] Figure 2 Showing the RNA-seq analysis results of DRG tissues in the AEW model.

[0020] Figure 3 Showing the real-time PCR analysis results of the Vgf expression level in the AEW model.

[0021] Figure 4 Showing the real-time PCR analysis results of the Vgf expression level in the AD model.

[0022] Figure 5 Showing the number of scratching times of C57BL / 6J mice after administration of TLQP-21.

[0023] Figure 6 Showing the number of scratching times of mast cell-deficient mice after administration of TLQP-21.

[0024] Figure 7 Showing the ameliorating effect on scratching in AD model mice by administration of a C3a receptor antagonist.

[0025] Figure 8 Showing the ameliorating effect on scratching in AEW model mice by administration of a C3a receptor antagonist.

[0026] Figure 9 Showing the ameliorating effect on scratching in AD model mice by montelukast.

[0027] Figure 10 Showing the ameliorating effect on scratching in ANIT model mice by montelukast. DETAILED DESCRIPTION OF THE INVENTION

[0028] The present inventors first collected dorsal root ganglia (DRGs) from model mice showing xerosis-like skin symptoms (hereinafter referred to as AEW model mice) using an acetone-ether mixture and water treatment, and comprehensively performed gene expression analysis. As a result, Vgf was identified as a gene showing expression changes. Furthermore, quantitative expression analysis of the Vgf gene in the DRGs of AEW model mice and atopic dermatitis model mice (hereinafter referred to as AD model mice) was performed. As a result, compared with control mice, Vgf expression in the DRG tissue was significantly increased in both model mice (Reference Example 1). Vgf is a gene encoding a neurosecretory factor (VGF nerve growth factor inducible, hereinafter referred to as VGF). It is known that VGF is decomposed by proteases in vivo to produce several bioactive peptides (Non-Patent Document 5). If TLQP-21, one of them, is injected into the skin of the nape of the neck of healthy mice, scratching behavior will occur in the mice, and it has been found that TLQP-21 induces itching (Reference Example 2). The scratching behavior caused by intradermal administration of this TLQP-21 also appears in mast cell-deficient mice, indicating that the induction of itching by TLQP-21 is a mast cell-independent response (Reference Example 3).

[0029] It is known that TLQP-21 binds to the complement factor C3a receptor, which is a GPCR-type receptor present on the cell membrane, and generates a signal (Non-Patent Document 6). Therefore, if a compound antagonistic to the C3a receptor is continuously administered to mice in parallel during the production of AD model mice and AEW model mice, it was confirmed that the scratching behavior was significantly reduced. From this, it was found that the C3a receptor becomes a target for suppressing itching, and by blocking the binding of TLQP-21 to the C3a receptor, itching can be suppressed (Reference Examples 4 and 5). It is considered that suppressing itching by blocking this C3a receptor signal transduction pathway is useful as a new technique for eliminating itching conditions that do not work with existing methods.

[0030] Therefore, the present invention relates to providing a novel raw material useful for preventing or improving itching by blocking the C3a receptor signal transduction pathway.

[0031] The present inventors studied effective raw materials for blocking the C3a receptor signal transduction pathway and found that montelukast has C3a receptor antagonistic activity.

[0032] So far, no reports have been made on the effects of montelukast on the C3a receptor and itching.

[0033] According to the present invention, by blocking the C3a receptor signal transduction pathway, itching and pruritic skin diseases can be prevented or improved.

[0034] In the present invention, the terutroban used is 4-[6-acetyl-3-[3-(4-acetyl-3-hydroxy-2-propylphenylthio)propoxy]-2-propylphenoxy]butyric acid (CAS Registry Number: 125961-82-2) represented by the following formula.

[0035]

[0036] Examples of pharmaceutically acceptable salts of terutroban include: alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salt; magnesium salt; aluminum salt; ammonium salt; organic amine salts such as trimethylamine salt, triethylamine salt, dicyclohexylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, procaine salt, N,N'-dibenzylethylenediamine salt, choline salt, ethylenediamine salt, meglumine (N-methylglucamine) salt, phenylethylbenzylamine (N-benzylphenethylamine) salt, diethylamine salt, piperazine salt, tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol) salt, or acid addition salts such as hydrochloride, hydrobromide, sulfate, acid sulfate, phosphate, acid phosphate, dihydrogen phosphate, acetate, succinate, citrate, mesylate (methanesulfonate), tosylate (p-toluenesulfonate).

[0037] In addition, terutroban can also be made into a prodrug. After the prodrug is administered to a living body, it becomes a pharmaceutically active compound through the action of enzymes or metabolic hydrolysis, etc. The prodrug only needs to be an acid derivative known to those skilled in the art. For example, esters produced by the reaction of terutroban with appropriate alcohols or amides produced by the reaction of terutroban with appropriate amines can be cited, and there is no particular limitation.

[0038] The terutroban, its pharmaceutically acceptable salt, or its prodrug can be a solvate (such as a hydrate, etc.) or a non-solvate.

[0039] Terutroban, its pharmaceutically acceptable salt, or its prodrug is a known compound and can be manufactured by known methods (Patent Documents 1 and 2 above) or can be obtained by purchasing commercially available products.

[0040] As shown in the following examples, terutroban shows C3a receptor antagonistic activity ( Figure 1 ). In addition, in AD model mice and pruritus model mice accompanied by cholestasis (ANIT model mice) administered with terutroban, it was confirmed that scratching behavior was significantly reduced ( Figure 9 and 10)。The C3a receptor is the receptor for C3a and TLQP-21 generated upon activation of the complement system. It has been reported that inhibiting the C3a receptor signaling pathway can suppress brain edema and intracerebral hemorrhage caused by stroke (PLoS One. 2017 Jul 10; 12(7): e0180822), and alleviate IgG-induced arthritis (J Pharmacol Sci. 2010; 112(1): 56-63).

[0041] Furthermore, as shown in Reference Example 2, when TLQP-21 generated by the decomposition of VGF is injected into the skin of the nape of the neck of healthy mice, scratching behavior is induced in the mice ( Figure 5 ). Based on this result, it was clarified that TLQP-21 induces itching. Scratching behavior generated by intradermal administration of TLQP-21 also appears in mast cell-deficient mice ( Figure 6 ), and thus, it was taught that the induction of itching by TLQP-21 is a mast cell-independent response.

[0042] As described above, it is known that TLQP-21 binds to the C3a receptor and generates a signal. Also, a known compound 1 that antagonizes the receptor for C3a was continuously administered to mice during the production of AD model mice and AEW model mice, and as a result, it was confirmed that scratching behavior was significantly reduced in the model mice administered with compound 1 ( Figure 7 and Figure 8 ). The reduction in this scratching behavior indicates the alleviation of itching. Therefore, it was clarified that the C3a receptor becomes a target for suppressing itching, particularly intractable itching.

[0043] Therefore, by using montelukast to antagonistically inhibit the binding of TLQP-21 to the C3a receptor, itching can be prevented or improved, and pruritic skin diseases can be prevented or improved.

[0044] That is, montelukast, its pharmaceutically acceptable salt, or its prodrug (hereinafter, also referred to as "the compound of the present invention") can become a C3a receptor antagonist, an agent for preventing or improving itching, or an agent for preventing or improving pruritic skin diseases (hereinafter, also referred to as "C3a receptor antagonist, etc."), and can be used for their use, etc. In addition, the compound of the present invention can be applied to animals including humans for use in inhibiting the C3a receptor signaling pathway, preventing or improving itching, and preventing or improving pruritic skin diseases.

[0045] Here, "use" can be a use in humans or non-human animals, and can be a therapeutic use or a non-therapeutic use. "Non-therapeutic" is a concept that does not include medical acts, that is, a concept that does not include methods of performing surgery, treatment, or diagnosis on the human body. More specifically, it is a concept that does not include methods of performing surgery, treatment, or diagnosis on the human body by a doctor or a person instructed by a doctor.

[0046] In the present specification, "C3a receptor antagonism" means hindering the action of TLQP-21 via the C3a receptor, for example, including hindering the C3a receptor signal transduction pathway.

[0047] "Itching" is a subjective feeling, and its cause is not particularly limited. The itchy part can be exemplified by a wide range or a specific part such as the whole body, scalp, face, back, wrist, fingernail, finger, foot, etc.

[0048] In the present invention, it is suitable for preventing or improving intractable itching.

[0049] "Intractable itching" means itching that cannot be resolved by antihistamines (H1 receptor antagonists). Examples of intractable itching include itching in atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, pruritus (for example, pruritus associated with chronic liver disorders, cholestasis, chronic kidney disorders and their dialysis treatments [hemodialysis or peritoneal dialysis], senile pruritus, winter pruritus, etc.), malignant neoplasms, and other diseases. Among them, itching in atopic dermatitis, itching in xeroderma, itching in nodular prurigo, or pruritus associated with cholestasis is preferred.

[0050] "Pruritic skin diseases" means skin diseases accompanied by itching. Examples include urticaria, atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, etc. Among these, skin diseases other than urticaria are skin diseases belonging to skin diseases accompanied by intractable itching. In the present invention, it is suitable for skin diseases accompanied by the above-mentioned intractable itching. Pruritic skin diseases are preferably atopic dermatitis, xeroderma, or nodular prurigo.

[0051] In addition, in the present specification, "prevention" means preventing or delaying the onset of symptoms in an individual, or reducing the risk of onset of symptoms in an individual.

[0052] "Improvement" means reversing, preventing, or delaying the progression of symptoms or conditions to a more severe state.

[0053] The C3a receptor antagonist of the present invention and the like can themselves be pharmaceuticals, quasi-pharmaceuticals, cosmetics, foods or feeds for blocking the C3a receptor signal transduction pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases, or can also be raw materials or preparations used in combination with such pharmaceuticals, quasi-pharmaceuticals, cosmetics, foods or feeds.

[0054] The pharmaceutical (including quasi-pharmaceutical, the same hereinafter) contains the compound of the present invention as an active ingredient for blocking the C3a receptor signal transduction pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases. Furthermore, the pharmaceutical may also contain a pharmaceutically acceptable carrier or other active ingredients, medicinal ingredients, etc., as needed, as long as the function of the active ingredient is not lost.

[0055] The administration method of the pharmaceutical containing the compound of the present invention is arbitrary, and examples include oral administration or parenteral administration. As dosage forms for oral administration, examples include tablets, capsules, granules, powders, syrups, etc. As dosage forms for parenteral administration, examples include various preparations such as topical skin use, transdermal, transmucosal, nasal, enteral, injection, suppository, inhalation, and patch. In the case of parenteral administration, a suitable dosage form is a topical skin preparation. Specifically, examples include forms such as ointments, emulsions, creams, lotions, washes, gels, and aerosols.

[0056] The cosmetic contains the compound of the present invention as an active ingredient for blocking the C3a receptor signal transduction pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases. Furthermore, the cosmetic may also contain a carrier or other active ingredients, cosmetic ingredients, etc. permitted in cosmetics, as needed, as long as the function of the active ingredient is not lost.

[0057] As a preferred example of the cosmetic containing the compound of the present invention, examples include cosmetics for the face and body (such as lotions, gels, creams, masks, etc.), cosmetics for makeup, cleansers for the face or body, etc.

[0058] Each preparation of the pharmaceutical or cosmetic can combine the compound of the present invention with a pharmaceutically or cosmetically acceptable carrier, the above-mentioned other active ingredients, medicinal ingredients, cosmetic ingredients, etc., as needed, and can be manufactured according to conventional methods.

[0059] As the pharmaceutically or cosmetically acceptable carrier, for example, examples include excipients, diluents, binders, disintegrants, coating agents, solubilizers, lubricant agents, glidants, solubilization aids, lubricants, various oils, surfactants, gelling agents, pH buffers, isotonic agents, preservatives, antioxidants, solvents, dispersants, chelating agents, thickeners, stabilizers, pH regulators, pigments, flavors, etc.

[0060] As such other active ingredients, pharmaceutical ingredients, or cosmetic ingredients, for example, plant extracts, bactericides, humectants, anti-inflammatory agents, antibacterial agents, keratolytic agents, ultraviolet absorbers, cooling agents, anti-seborrheic agents, detergents, makeup ingredients, etc. can be cited.

[0061] The food contains the compound of the present invention as an active ingredient for blocking the C3a receptor signaling pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases.

[0062] Food includes foods (such as foods for specified health use, foods with functional labeling, foods for special purposes, etc.) that require the prevention or improvement of itching, or the prevention or improvement of pruritic skin diseases, and have the indication approved or declared as such. As an example of the indication, there is "improve skin discomfort (itching)". Foods with approved or declared functional labeling can be distinguished from ordinary foods.

[0063] The form of the food can be solid, semi-solid, or liquid (such as beverages). As examples, various food compositions (breads, cakes, noodles, snacks, frozen foods, ice creams, sugars, seasoned flakes, soups, dairy products, milkshakes, beverages, seasonings, etc.) can be cited, and further nutritional supplement compositions in the same forms as the above oral administration preparations (solid preparations such as granules, powders, tablets, capsules, microcapsules, lozenges, etc.).

[0064] Foods in various forms can appropriately combine the compound of the present invention with any food materials or other active ingredients, or additives permitted in foods (such as solvents, softeners, oils, emulsifiers, preservatives, acidulants, sweeteners, bitter agents, pH regulators, stabilizers, colorants, ultraviolet absorbers, antioxidants, humectants, thickeners, fixatives, dispersants, flow improvers, wetting agents, fragrances, seasonings, flavor regulators), etc., and be prepared according to convention.

[0065] The feed contains the compound of the present invention as an active ingredient for blocking the C3a receptor signaling pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases.

[0066] As the form of the feed, it is preferably granular, flaky, pasty, or liquid. For example, livestock feeds for cows, pigs, chickens, sheep, horses, etc., small animal feeds for rabbits, rats, mice, etc., and pet foods for dogs, cats, birds, etc. can be cited.

[0067] The feed can appropriately combine the compound of the present invention with other feed materials, such as meats, proteins, grains, bran, meal, sugars, vegetables, vitamins, minerals, gelling agents, shaping agents, pH regulators, seasonings, preservatives, nutritional enhancers, etc., and be prepared according to convention.

[0068] The content of the compound of the present invention in the above-mentioned preparation varies depending on the form of the preparation and the like, and cannot be generalized. However, based on the total amount of the preparation, in terms of montelukast conversion, it is preferably 0.001% by mass or more, more preferably 0.005% by mass or more. In addition, it is preferably 80% by mass or less, more preferably 40% by mass or less. In addition, it is preferably 0.001 to 80% by mass, more preferably 0.005 to 40% by mass.

[0069] The dosage or usage amount of the compound of the present invention may be an amount that can achieve the effects of the present invention. This dosage or usage amount may vary depending on the type, body weight, gender, age, condition, or other factors of the subject. In the case of oral administration such as tablets or capsules, in terms of montelukast, it is preferably 0.005 mg or more, more preferably 1 mg or more per adult (60 kg) each time. In addition, it is preferably 3000 mg or less, more preferably 100 mg or less. In addition, it is preferably 0.005 mg to 3000 mg, more preferably 1 mg to 100 mg. In the present invention, this amount can be divided into 1 to multiple times within a day, and administered or used repeatedly and continuously for 1 day or more, preferably 7 days or more, more preferably 14 days or more, and even more preferably 42 days or more.

[0070] Examples of the subject for administering or using the C3a receptor antagonist of the present invention include humans or non-human animals that require or expect to block the C3a receptor signaling pathway, prevent or improve itching, or prevent or improve pruritic skin diseases. Specifically, examples include humans or non-human animals with refractory itching or suffering from pruritic skin diseases. Examples of non-human animals include non-human mammals such as anthropoid apes, other primates, and felines.

[0071] In addition, as the site for administering or using the C3a receptor antagonist of the present invention, in the case of parenteral administration such as a topical skin preparation, it may be any itchy site, and there is no particular limitation.

[0072] Regarding the above-mentioned embodiments, the present invention further discloses the following embodiments.

[0073] <1> A C3a receptor antagonist, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

[0074] <2> A preventive or ameliorating agent for itching, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

[0075] <3> A preventive or ameliorating agent for pruritic skin diseases, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

[0076] <4>The itching prevention or improvement agent as described in <2>, wherein the itching is preferably intractable itching, more preferably itching in diseases such as atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, pruritus (e.g., pruritus associated with chronic liver disorders, pruritus associated with cholestasis, pruritus associated with chronic renal disorders and their dialysis treatments [hemodialysis or peritoneal dialysis], senile pruritus, winter pruritus, etc.), malignant neoplasms, etc., and further preferably itching in atopic dermatitis, itching in xeroderma, itching in nodular prurigo, or pruritus associated with cholestasis.

[0077] <5>The itching skin disease prevention or improvement agent as described in <3>, wherein the itching skin disease is preferably urticaria, atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, and more preferably atopic dermatitis, xeroderma or nodular prurigo.

[0078] <6>The agent as described in any one of <1> to <5>, wherein the content of montelukast, its pharmaceutically acceptable salt, or its prodrug in the preparation is preferably 0.001% by mass or more, more preferably 0.005% by mass or more, based on the total amount of the preparation, and is preferably 80% by mass or less, more preferably 40% by mass or less, and is preferably 0.001 to 80% by mass, more preferably 0.005 to 40% by mass, in terms of montelukast conversion.

[0079] <7>The agent as described in any one of <1> to <6>, wherein it is a pharmaceutical preparation.

[0080] <8>The dosage or usage amount of montelukast, its pharmaceutically acceptable salt, or its prodrug, when administered orally, is preferably 0.005 mg or more, more preferably 1 mg or more, per adult (60 kg) each time, and is preferably 3000 mg or less, more preferably 100 mg or less, and is preferably 0.005 mg to 3000 mg, more preferably 1 mg to 100 mg, in terms of montelukast.

[0081] <9>The use of montelukast, its pharmaceutically acceptable salt, or its prodrug in the manufacture of a C3a receptor antagonist.

[0082] <10>Montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, which is used for C3a receptor antagonism.

[0083] <11>Non-therapeutic use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof for C3a receptor antagonism.

[0084] <12>A method for C3a receptor antagonism, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is administered or applied to a subject.

[0085] <13>Use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the manufacture of an agent for preventing or improving itching.

[0086] <14>Montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, which is used for preventing or improving itching.

[0087] <15>Non-therapeutic use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof for preventing or improving itching.

[0088] <16>A method for preventing or improving itching, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is administered or applied to a subject.

[0089] <17>The use as described in <13>, the montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof as described in <14>, the non-therapeutic use as described in <15>, or the method as described in <16>, wherein the itching is preferably intractable itching, more preferably itching in atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, pruritus (e.g., pruritus associated with chronic liver disorders, cholestasis, chronic kidney disorders and their dialysis treatments [hemodialysis or peritoneal dialysis], senile pruritus, winter pruritus, etc.), malignant neoplasms and other diseases, and further preferably itching in atopic dermatitis, itching in xeroderma, itching in nodular prurigo, or pruritus associated with cholestasis.

[0090] <18>Use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the manufacture of an agent for preventing or improving pruritic skin diseases.

[0091] <19>Montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, which is used for preventing or improving pruritic skin diseases.

[0092] <20>The non-therapeutic use of terutroban, its pharmaceutically acceptable salts, or its prodrugs in the prevention or improvement of pruritic skin diseases.

[0093] <21>A method for preventing or improving a pruritic skin disease, wherein terutroban, its pharmaceutically acceptable salts, or its prodrugs are administered or applied to a subject.

[0094] <22>The use according to <18>, terutroban according to <19>, its pharmaceutically acceptable salts, or its prodrugs, the non-therapeutic use according to <20>, the method according to <21>, wherein the pruritic skin disease is preferably urticaria, atopic dermatitis, xeroderma (including senile xeroderma, asteatotic dermatitis, asteatotic eczema), contact dermatitis, seborrheic dermatitis, nummular eczema, psoriasis, prurigo, nodular prurigo, chronic prurigo, pemphigoid, dermatomyositis, more preferably atopic dermatitis, xeroderma or nodular prurigo.

[0095] [Examples]

[0096] Example 1 C3a receptor antagonistic effect of terutroban

[0097] 1. C3a receptor ligand

[0098] Mouse TLQP-21 (Tocris Bioscience, sequence: TLQPPASSRRRHFHHALPPAR, the same hereinafter) and human C3a recombinant protein (R&D systems) were used. The solvent was set as HBSS (Gibco).

[0099] 2. Evaluation samples

[0100] As C3a receptor antagonists (antagonists), compound 1 (L-arginine, N2-[[5-(diphenylmethyl)-2-thienyl]carbonyl]) synthesized based on existing reports (Rowley, J.A. et al. Journal of Medicinal Chemistry, 2020; 63(2):529-541) and terutroban (Cayman Chemical) were used. At the following evaluation, compound 1 and terutroban were dissolved with DMSO and appropriately diluted with HBSS.

[0101] 3. Cells for C3a receptor response evaluation

[0102] Human embryonic kidney cells 293 (HEK293) into which the expression vector required for evaluation was introduced according to the following contents were used. Human C3a receptor expression vector (OriGene), GNA16 expression vector, and PEI-MAX (PolyScience) solution were added to serum-free DMEM (Dulbecco's Modified Eagle Medium) culture medium in sequence and mixed thoroughly, and then allowed to stand at room temperature for 20 minutes to prepare a transfection solution. The prepared transfection solution was added to HEK293 seeded in a 6-well cell culture plate, and the expression vector was introduced into HEK293 after culturing for more than 24 hours. After culturing, the culture medium was removed, and the cells were washed once with PBS, and the cells were detached with 0.05% trypsin / EDTA (Gibco) and supplied to the following Ca 2+ medium. 2+ Flow detection (Ca 2+ -flux assay). The human GNA16 expression vector was prepared by inserting a sequence identical to the cDNA sequence of human GNA16 (Accession No. M63904.1) from the base sequence database (GenBank) provided by the National Center for Biotechnology Information of the United States into a plasmid vector pIRESneo3 (manufactured by Clontech).

[0103] 4. Ca 2+ Flow Detection

[0104] The cells for evaluating the C3a receptor response were seeded in BioCoat (registered trademark) Poly-D-Lysine 96-well Black Flat Bottom Microplate (CORNING). The intracellular Ca2+ upon C3a receptor stimulation was measured using the Fluo-4 Calcium Kit (DOJINDO). 2+ Add the loading buffer prepared according to the manual of the kit and let it stand at 37°C for 1 hour to increase the Ca concentration. 2+The fluorescent indicator Fluo-4 was added to the cells. After washing once with HBSS, a recording buffer containing compound 1 or montelukast, which is a C3a receptor antagonist, was added. Furthermore, a recording buffer was added to the control. After 15 minutes, a C3a receptor ligand solution (mouse TLQP-21 solution or human C3a recombinant protein solution) was added for C3a receptor stimulation, and the Ca 2+ influx associated with the stimulation was measured over time with intracellular fluorescence intensity (Ex: 480 nm, Em: 540 nm). The value obtained by subtracting the measured value (blank) obtained by adding HBSS instead of the C3a receptor ligand solution was used as the measured value. Furthermore, the compound 1 solution, montelukast, mouse TLQP-21, and human C3a recombinant protein solution were appropriately diluted so that the final concentrations at the time of measuring the fluorescence intensity were 1 μM and 10 μM, 1 μM, and 300 nM, respectively, and the concentrations were adjusted and added. The addition of the C3a receptor antagonist solution and the C3a receptor ligand solution to the cells for C3a receptor response evaluation and the measurement of the fluorescence intensity were performed using a fluorescence microplate reader (FDSS / μCELL, manufactured by Hamamatsu Photonics K.K.) equipped with an automatic dispensing function, and were carried out for 3 minutes over time after adding the C3a receptor ligand solution. When the integrated value of the fluorescence intensity measured in the control was set to 100%, the results obtained by calculating the cumulative value (%) of the fluorescence intensity of montelukast are shown in Figure 1 .

[0105] As Figure 1 shown, the fluorescence intensity generated by C3a receptor stimulation by the C3a receptor ligand measured in the control was inhibited by adding compound 1 or montelukast, indicating that montelukast has the effect of a C3a receptor antagonist.

[0106] Reference Example 1 Comprehensive Gene Expression Analysis

[0107] 1. Experimental Animals

[0108] Male C57BL / 6J mice and male NC / Nga mice were used.

[0109] Xerosis model (AEW model) : Absorbent cotton soaked in a 1:1 (v / v) mixture of acetone and diethyl ether was pressed against the skin of the shaved C57BL / 6J mice and left for 15 seconds. Then, immediately, absorbent cotton soaked in water was pressed for 30 seconds. This treatment was repeated 2 times a day (morning and evening) for 7 days to produce AEW model mice. As a control, control mice were produced by pressing absorbent cotton soaked in water for 30 seconds only. n = 7 for each group was used for this analysis.

[0110] Atopic dermatitis model (AD model):Infest NC / Nga mice with the mouse louse (M. musculi) to spontaneously develop dermatitis, thereby establishing an AD model. As control mice, use mice of this system that are not infested with mites and are raised under SPF conditions. n = 8 mice were used for each group in this analysis.

[0111] 2. Purification of total RNA

[0112] Dissect the dorsal root ganglia from the cervical vertebrae of the mice. Homogenize the dissected tissues using a Polytron Homogenizer, and purify the total RNA using the RNeasy Mini Kit from QIAGEN.

[0113] 3. Gene expression analysis

[0114] RNA-seq : Use the total RNA purified from the AEW model mice and their control mice. For reverse transcription, use SuperScript VILO (Thermo fisher scientific), and subsequent processing is based on the Ion AmpliSeq standard protocol from Thermo fisher scientific. The sequencing is performed using the Ion S5 system. The results are shown in Figure 2 below.

[0115] As Figure 2 shown, in the AEW model, by Student's t-test, p < 0.05, compared with the control mice, the expression of Vgf in each fixed amount of sequence was significantly increased

[0116] Real-time PCR : Use the total RNA purified from the AEW model mice, AD model mice, and their control mice. Using Vgf as the target gene, perform quantitative PCR analysis of the Vgf gene by a Real-Time PCR System using specific TaqMan Gene Expression Assays. As an internal standard, use Rplp0. The relative expression levels of the Vgf gene, with the Vgf gene expression level of the control mice set to 1, are shown in Figure 3 and Figure 4 below.

[0117] As Figure 3 shown, in the AEW model mice, the expression level of Vgf in the DRG tissue was significantly increased compared with the control mice. Additionally, as Figure 4 shown, in the AD model mice, the expression level of Vgf in the DRG tissue was also significantly increased compared with the control mice.

[0118] Itching manifestation by TLQP-21 in Reference Example 2

[0119] 1. Experimental animals

[0120] Male C57BL / 6J mice were used as the mice.

[0121] 2. Administration of mediator candidate molecules

[0122] Mouse TLQP-21 (Tocris Bioscience) was prepared to be 15 nmol / 20 μL and 30 nmol / 20 μL. The solvent was set as physiological saline. 20 μL of it was administered intradermally to the nape of the shaved C57BL / 6J mice. Immediately after injection, the scratching behavior of the mice was measured, and the measured values 30 minutes after the start were analyzed.

[0123] 3. Measurement of scratching behavior

[0124] The scratching behavior after administration of TLQP-21 was measured using MicroAct (Neuroscience). The results are shown in Figure 5 . The total value of consecutive single scratching actions was recorded as Events, and the total value of the number of scratching actions among them was recorded as Beats.

[0125] As Figure 5 shown, after administration of TLQP-21, an increase in the scratching behavior of the mice was observed in the form of a significant increase in Events and Beats. The manifestation of the scratching behavior was an immediate response within 30 minutes.

[0126] Itching manifestation dependent on TLQP-21 in mast cell-deficient mice in Reference Example 3

[0127] 1. Experimental animals

[0128] Mice using the WBB6F1 / Kit- Kit W / Kit W-v system as mast cell-deficient mice and the WBB6F1+ / + system as wild-type control mice were used. Male mice were used in both cases.

[0129] 2. Administration of mediator molecules

[0130] Mouse TLQP-21 (Tocris Bioscience) was prepared to be 20 nmol / 20 μL. The solvent was set as physiological saline. 20 μL of it was administered intradermally to the nape of the shaved WBB6F1 / Kit- Kit W / Kit W - v system and WBB6F1+ / + system. Immediately after injection, the scratching behavior of the mice was measured, and the measured values 30 minutes after the start were analyzed.

[0131] 3. Measurement of scratching behavior

[0132] The scratching behavior after administration of TLQP-21 was measured using MicroAct (Neuroscience). The results are shown in Figure 6 . The total value of consecutive single scratching actions was recorded as an event.

[0133] As Figure 6 shown, an increase in the scratching behavior of mice caused by administration of TLQP-21 was observed in either system of mast cell-deficient mice and wild-type control mice.

[0134] Improvement of itching in AD model mice by the C3a receptor antagonist in Reference Example 4

[0135] 1. Experimental animals

[0136] Seven-week-old female NC / Nga mice were used.

[0137] Preparation of AD model mice : The hair on the nape of the neck was shaved, and under isoflurane anesthesia, 100 mg of mite allergen-containing ointment: Biostir AD (Biostir) was applied to the nape of the neck and auricles for initial sensitization. Four days after the first application of the mite allergen-containing ointment, 150 μL of 4% (w / v) SDS aqueous solution and 100 mg of the mite allergen-containing ointment were applied under isoflurane anesthesia. Thereafter, the same operation was performed 4 times at a frequency of once every 3 - 4 days to induce AD-like dermatitis.

[0138] 2. Sample preparation and administration method

[0139] As the C3a receptor antagonist (antagonist), Compound 1 that was confirmed to have a C3a receptor antagonistic effect in Example 1 above was used. Compound 1 was dissolved in physiological saline containing 20% polyethylene glycol 400, and 200 μL was subcutaneously administered to the nape of the neck. Only the same amount of the above solvent was administered to the control group. The concentration of the C3a receptor antagonist (antagonist) solution was set at 0.5 mg / mL. Sample administration was started on the day of the second application of the mite allergen-containing ointment and was performed 3 times a week.

[0140] 3. Measurement of scratching behavior

[0141] Four days after the last application of the mite allergen-containing ointment, the scratching behavior was measured for 3 hours after the final administration of the sample. The scratching behavior was measured using MicroAct (Neuroscience). The results are shown in Figure 7 .

[0142] As Figure 7As shown, in AD model mice administered with compound 1, which was confirmed to have C3a receptor antagonistic activity, in parallel with Dermatophagoides farinae allergen, it was confirmed that scratching behavior was significantly reduced compared to AD model mice administered only with the solvent.

[0143] Improvement of itching in AEW model mice by the C3a receptor antagonist in Reference Example 5

[0144] 1. Experimental animals

[0145] Male C57BL / 6J mice were used as animals.

[0146] Preparation of AEW model : The hair on the nape of the neck was shaved, and absorbent cotton soaked in a 1:1 (v / v) mixture of acetone and diethyl ether was pressed onto the nape of the neck and left for 15 seconds. Immediately afterwards, absorbent cotton soaked in water was pressed for 30 seconds. This treatment was repeated 2 times a day (morning and evening) for 7 days.

[0147] 2. Sample preparation and administration method

[0148] As the C3a receptor antagonist (antagonist), compound 1, which was confirmed to have C3a receptor antagonistic activity in Example 1 above, was used. Compound 1 was dissolved in physiological saline containing 20% polyethylene glycol 400, and 200 μL was subcutaneously administered to the nape of the neck. Only the same amount of the above solvent was administered to the control group. The concentration of the C3a receptor antagonist (antagonist) solution was set at 0.5 mg / mL. Sample administration started after the first AEW treatment and was carried out once a day for 8 days.

[0149] 3. Measurement of scratching behavior

[0150] Sixteen hours after the last AEW treatment, scratching behavior was measured for 3 hours after the final administration of the sample. MicroAct (Neuroscience) was used to measure scratching behavior. The results are shown in Figure 8 in.

[0151] As Figure 8 shown, in AEW model mice administered with compound 1, which was confirmed to have C3a receptor antagonistic activity, in parallel with AEW treatment, it was confirmed that scratching behavior was significantly reduced compared to AEW model mice administered only with the solvent.

[0152] Improvement of itching in AD model mice by terutroban in Example 2

[0153] 1. Experimental animals

[0154] M. musculi was parasitized on NC / Nga mice to spontaneously induce dermatitis, thereby establishing an AD model. Male mice at 14 weeks of age with dermatitis were used. After 1 week of acclimation, hair on the nape of the neck was shaved and the scratching behavior was measured, and the mice were divided into 2 groups of 8 each in such a way that the number of scratching behaviors was equal.

[0155] 2. Preparation of evaluation samples

[0156] Montelukast (Cayman Chemical) was used. In addition, carboxymethyl cellulose (Maruishi Pharmaceutical Co., Ltd.) prepared as 0.5% with water for injection was used as a solvent. The weighed montelukast was gently crushed using an agate mortar and pestle, and then the solvent was added dropwise and mixed repeatedly to prepare a suspension (5 mg / mL). It was prepared on the day of administration and stored at room temperature until the period of administration.

[0157] 3. Administration of evaluation samples

[0158] The body weight was measured, the dosage (10 mL / kg) was calculated, and the prepared evaluation samples were administered orally using a gastric tube and a disposable syringe for mice.

[0159] 4. Measurement of scratching behavior

[0160] MicroAct (Neuroscience) was used to measure the scratching behavior. The number of scratching behaviors was set as the total value (events) of consecutive single scratching actions, and the measurement time was set as 22 hours. It was measured on the day before administration of montelukast (before administration) and on the day of administration (after administration), and the number of scratching behaviors before and after administration was compared to evaluate the antipruritic effect of montelukast. The results are shown in Figure 9 below.

[0161] As Figure 9 shown, it was confirmed that after administration of montelukast, the scratching behavior in AD model mice was significantly reduced.

[0162] Example 3 Improvement of itching in cholestasis-associated pruritus model mice (ANIT model mice) caused by montelukast.

[0163] 1. Experimental animals

[0164] Use shaved male C57BL / 6J mice. Alpha-naphthyl isothiocyanate (ANIT) dissolved in olive oil (FUJIFILM Wako Pure Chemical Corporation) was orally administered once a day to 8-week-old C57BL / 6J mice for 10 days using a mouse gavage tube and a disposable syringe. At the time of administration, the body weight was measured in advance, and the administration was carried out in such a way as to be 25 mg / 10 mL / kg (ANIT model). It has been reported that mice orally administered ANIT for several consecutive days showed cholestasis-like symptoms and an increase in scratching behavior (eLife, 2019 8:e44116.).

[0165] 2. Preparation of evaluation samples

[0166] Use zileuton (Cayman Chemical). In addition, a solvent prepared by making carboxymethyl cellulose (Maruishi Pharmaceutical Co., Ltd.) into 0.5% with water for injection was used. The weighed zileuton was gently crushed using an agate mortar and pestle, and then the solvent was added dropwise and mixed repeatedly to prepare a suspension (5 mg / mL). It was prepared on the day of administration and stored at room temperature until the administration period.

[0167] 3. Administration of evaluation samples

[0168] Measure the body weight, calculate the dose (10 mL / kg), and orally administer the prepared evaluation sample using a mouse gavage tube and a disposable syringe.

[0169] 4. Measurement of scratching behavior

[0170] Use MicroAct (Neuroscience) to measure scratching behavior. The number of scratching behaviors was set as the total value (events) of consecutive single scratching actions, and the measurement time was set to 22 hours. It was measured on the day before the administration of zileuton (before administration) and on the day of administration (after administration), and the number of scratching behaviors before and after administration was compared to evaluate the antipruritic effect of zileuton. The results are shown in Figure 10 below.

[0171] As Figure 10 shown, it was confirmed that after the administration of zileuton, the scratching behavior in ANIT model mice was significantly reduced.

Claims

1. A C3a receptor antagonist, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

2. An agent for preventing or improving itching, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

3. The agent for preventing or improving itching according to claim 2, wherein the itching is intractable itching.

4. The agent for preventing or improving itching according to claim 2 or 3, wherein the itching is itching in atopic dermatitis, itching in xerosis, itching in prurigo nodularis, or pruritus associated with cholestasis.

5. An agent for preventing or improving pruritic skin diseases, wherein montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof is used as an active ingredient.

6. The agent for preventing or improving pruritic skin diseases according to claim 5, wherein the pruritic skin diseases are atopic dermatitis, xerosis, or prurigo nodularis.

7. Use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the manufacture of a C3a receptor antagonist.

8. Use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the manufacture of an agent for preventing or improving itching.

9. The use according to claim 8, wherein the itching is intractable itching.

10. The use according to claim 8 or 9, wherein the itching is itching in atopic dermatitis, itching in xerosis, itching in prurigo nodularis, or pruritus associated with cholestasis.

11. Use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the manufacture of an agent for preventing or improving pruritic skin diseases.

12. The use according to claim 11, wherein the pruritic skin diseases are atopic dermatitis, xerosis, or prurigo nodularis.

13. A montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, wherein it is used for C3a receptor antagonism.

14. A montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, wherein it is used for preventing or improving itching.

15. The montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof according to claim 14, wherein the itching is intractable itching.

16. The montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof according to claim 14 or 15, wherein the itching is itching in atopic dermatitis, itching in xerosis, itching in prurigo nodularis, or pruritus associated with cholestasis.

17. A montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof, wherein it is used for preventing or improving pruritic skin diseases.

18. The montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof according to claim 17, wherein the pruritic skin diseases are atopic dermatitis, xerosis, or prurigo nodularis.

19. Non-therapeutic use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in C3a receptor antagonism.

20. Non-therapeutic use of montelukast, a pharmaceutically acceptable salt thereof, or a prodrug thereof in preventing or improving itching.

21. The non-therapeutic use according to claim 20, wherein the itching is intractable itching.

22. The non-therapeutic use according to claim 20 or 21, wherein the itching is itching in atopic dermatitis, itching in xeroderma, itching in prurigo nodularis, or pruritus accompanied by cholestasis.

23. The non-therapeutic use of terutroban, its pharmaceutically acceptable salt, or its prodrug for preventing or improving pruritic skin diseases.

24. The non-therapeutic use according to claim 23, wherein the pruritic skin disease is atopic dermatitis, xeroderma or prurigo nodularis.

25. A method for antagonizing C3a receptor, wherein terutroban, its pharmaceutically acceptable salt, or its prodrug is administered or applied to a subject.

26. A method for preventing or improving itching, wherein terutroban, its pharmaceutically acceptable salt, or its prodrug is administered or applied to a subject.

27. The method according to claim 26, wherein the itching is intractable itching.

28. The method according to claim 26 or 27, wherein the itching is itching in atopic dermatitis, itching in xeroderma, itching in prurigo nodularis, or pruritus accompanied by cholestasis.

29. A method for preventing or improving a pruritic skin disease, wherein terutroban, its pharmaceutically acceptable salt, or its prodrug is administered or applied to a subject.

30. The method according to claim 29, wherein the pruritic skin disease is atopic dermatitis, xeroderma or prurigo nodularis.

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