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 ozanimod, 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 trimmed 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 function to receive sensations in the skin and transmit them to the second-order neurons in the posterior horn of the spinal cord.

[0004] Itch-causing substances cause itching by binding to corresponding receptors. As itch-causing substances, histamine, serotonin, chloroquine, etc. have been reported, and as itch-enhancing substances (sensitizing substances), Th2 cytokines such as IL-4 or IL-13 have been reported. Among them, the representative chemical itch-causing 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 for suppressing 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 further 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, ozanimod is known as an oral agonist for subtypes 1 and 5 of sphingosine-1-phosphate (S1P) receptors (Patent Document 1), and it has been reported that ozanimod may reduce the ability of lymphocytes to exit lymph nodes, thereby reducing the number of circulating lymphocytes in peripheral blood. In Japan, ozanimod is being studied as an oral therapeutic drug for multiple sclerosis and inflammatory bowel disease.

[0006] (Patent Document 1) Japanese Patent Application Laid-Open No. 2011-523412

[0007] (Non-Patent Document 1) Hashimoto Y et al. Life Sciences. 2004 Dec 31; 76(7):783-94

[0008] (Non-Patent Document 2) Ikoma A et al. Nature Reviews Neuroscience. 2006 Jul; 7(7):535-47

[0009] (Non-Patent Document 3) J Am Acad Dermatol. 2014 Jul; 71(1):116-132

[0010] (Non-Patent Document 4) Future Oncol. 2018 Oct; 14(24):2531-2541

[0011] (Non-Patent Document 5) Lewis JE et al. Front Endocrinol (Lausanne). 2015 Feb 2; 6:3

[0012] (Non-Patent Document 6) Cero C et al Structure. 2014 Dec 2; 22(12):1744-1753 SUMMARY OF THE INVENTION

[0013] The present invention relates to the following 1) to 3).

[0014] 1) A C3a receptor antagonist, which contains ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

[0015] 2) An agent for preventing or improving itching, which contains ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient.

[0016] 3) An agent for preventing or improving pruritic skin diseases, which contains ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof as an active ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Indicates the C3a receptor antagonistic effect of ozanimod.

[0018] Figure 2 Indicates the RNA-seq analysis results of DRG tissues in the AEW model.

[0019] Figure 3 Indicates the real-time PCR analysis results of the Vgf expression level in the AEW model.

[0020] Figure 4 Indicates the real-time PCR analysis results of the Vgf expression level in the AD model.

[0021] Figure 5 Indicates the scratching frequency of C57BL / 6J mice after administration of TLQP-21.

[0022] Figure 6 Indicates the scratching frequency of mast cell-deficient mice after administration of TLQP-21.

[0023] Figure 7 Indicates the improvement effect of scratching in AD model mice by administration of a C3a receptor antagonist.

[0024] Figure 8 Indicates the improvement effect of scratching in AEW model mice by administration of a C3a receptor antagonist.

[0025] Figure 9 Indicates the improvement effect of scratching in AD model mice by ozanimod.

[0026] Figure 10 Indicates the improvement effect of scratching in ANIT model mice by ozanimod. Detailed implementation manners

[0027] 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 neuropeptide (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 posterior neck of healthy mice, scratching behavior will occur in the mice, and it is 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).

[0028] 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 determined 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 signaling pathway is useful as a new technique for eliminating itching conditions that do not respond to existing methods.

[0029] Therefore, the present invention relates to providing a novel raw material useful for preventing or improving itching by blocking the C3a receptor signaling pathway.

[0030] The present inventors studied effective raw materials for blocking the C3a receptor signaling pathway and found that ozanimod has C3a receptor antagonistic activity.

[0031] So far, no reports have been made on the effects of ozanimod on the C3a receptor and itching.

[0032] According to the present invention, by blocking the C3a receptor signaling pathway, itching and pruritic skin diseases can be prevented or improved.

[0033] In the present invention, the ozanimod used is 5-[3-[1-(2-hydroxyethylamino)-2,3-dihydro-1H-inden-4-yl]-1,2,4-oxadiazol-5-yl]-2-isopropoxybenzonitrile (CAS Registry Number: 1306760-87-1) represented by the following formula. There are optical isomers in ozanimod, which can be any optical isomer or a mixture of optical isomers.

[0034]

[0035] Examples of pharmaceutically acceptable salts of ozanimod 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, glucosamine (N-methylglucosamine) salt, phenethylbenzylamine (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, tosylate, etc.

[0036] In addition, ozanimod 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 ozanimod with appropriate alcohols or amides produced by the reaction of ozanimod with appropriate amines can be cited, and there is no particular limitation.

[0037] The ozanimod, its pharmaceutically acceptable salt, or its prodrug can be a solvate (such as a hydrate, etc.) or a non-solvate.

[0038] Ozanimod, its pharmaceutically acceptable salt, or its prodrug is a known compound and can be manufactured by known methods (Patent Document 1 above) or can be obtained by purchasing commercially available products.

[0039] As shown in the following examples, ozanimod 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 ozanimod, it was confirmed that the scratching behavior was significantly reduced ( Figure 9 and 10 ).

[0040] The C3a receptor is the receptor for C3a and TLQP-21 produced upon activation of the complement system. It has been reported that blocking the C3a receptor signaling pathway inhibits brain edema and cerebral hemorrhage caused by stroke (PloS One. 2017 Jul 10; 12(7): e0180822) and alleviates IgG-induced arthritis (J Pharmacol Sci. 2010; 112(1): 56-63).

[0041] Furthermore, as shown in Reference Example 2, when TLQP-21 produced by the decomposition of VGF was injected into the skin of the nape of the neck of healthy mice, scratching behavior was induced in the mice ( Figure 5 ). From this result, it was clarified that TLQP-21 induces itching. The scratching behavior produced by intradermal administration of TLQP-21 also occurred 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 compound 1 known to antagonize 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 in the model mice administered with compound 1, the scratching behavior was significantly reduced ( Figure 7 and Figure 8 ). The reduction of 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 ozanimod to antagonistically block 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, ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof (hereinafter, also referred to as "the compound of the present invention") can be 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 in the manufacture thereof. In addition, the compound of the present invention can be applied to animals including humans and used for blocking the C3a receptor signaling pathway, preventing or improving itching, and preventing or improving pruritic skin diseases.

[0045] Here, the "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, and more specifically, 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 sensation, and its cause is not particularly limited. The itchy part can be, for example, a wide range such as the whole body, scalp, face, back, wrists, fingernails, fingers, feet, etc., or a specific part.

[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, dry skin (including senile xerosis, 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 dry skin, 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, dry skin (including senile xerosis, 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, dry skin, 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 inhibiting the C3a receptor signaling 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 inhibiting the C3a receptor signaling pathway, for preventing or improving itching, and for preventing or improving pruritic skin diseases. Furthermore, the pharmaceutical may also contain, as required, a pharmaceutically acceptable carrier or other active ingredients, medicinal ingredients, etc., as long as the function of the active ingredient is not impaired.

[0055] The administration route 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, aerosols, etc.

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

[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 washes, 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 permitted carrier, the above-mentioned other active ingredients, medicinal ingredients, cosmetic ingredients, etc., as required, and can be manufactured according to conventional methods.

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

[0060] As such other active ingredients, pharmaceutical ingredients, or cosmetic ingredients, examples include plant extracts, bactericides, humectants, anti-inflammatory agents, antibacterial agents, keratolytics, ultraviolet absorbers, cooling agents, anti-seborrheic agents, detergents, and makeup ingredients.

[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] Foods include those that require the prevention or improvement of itching, or the prevention or improvement of pruritic skin diseases, and have a label indicating this as required and approved or declared as such (such as foods for specified health uses, foods with functional labeling, and foods for special purposes). As an example of a label, 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 a beverage). Examples include various food compositions (breads, cakes, noodles, snacks, frozen foods, ice creams, sugars, savory snacks, soups, dairy products, milkshakes, beverages, seasonings, etc.), and furthermore, nutritional supplement compositions in the same form as the above oral dosage forms (solid dosage forms 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, 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, fixing agents, dispersants, fluidity 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, paste-like, or liquid. Examples include livestock feeds for cattle, pigs, chickens, sheep, horses, etc., small animal feeds for rabbits, rats, mice, etc., and pet foods for dogs, cats, birds, etc.

[0067] The feed can be prepared by appropriately combining the compound of the present invention with other feed materials, such as meat, protein, grains, bran, meal, sugars, vegetables, vitamins, minerals, gelling agents, shape retainers, pH regulators, flavoring agents, preservatives, nutritional enhancers, etc., according to convention.

[0068] The content of the compound of the present invention in the above preparations varies depending on the form of the preparation, etc., and cannot be generalized. However, based on, for example, the total amount of the preparation, in terms of ozanimod conversion, it is preferably 0.01% by mass or more, more preferably 0.05% by mass or more. Additionally, it is preferably 1.2% by mass or less, more preferably 0.6% by mass or less. Further, it is preferably 0.01 - 1.2% by mass, more preferably 0.05 - 0.6% by mass.

[0069] The dosage or usage amount of the compound of the present invention can be an amount capable of achieving 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 ozanimod, for each adult (60 kg) each time, it is preferably 0.01 mg or more, more preferably 0.1 mg or more. Additionally, it is preferably 100 mg or less, more preferably 10 mg or less. Further, it is preferably 0.01 mg - 100 mg, more preferably 0.1 mg - 10 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] As the subject for administering or using the C3a receptor antagonist of the present invention, etc., examples can include humans or non - human animals that require or expect to block the C3a receptor signaling pathway, prevent or improve itching, and prevent or improve pruritic skin diseases. Specifically, examples can include humans or non - human animals with refractory itching or suffering from pruritic skin diseases. As non - human animals, examples can include non - human mammals such as anthropoid apes, other primates, and felines, etc.

[0071] Additionally, as the site for administering or using the C3a receptor antagonist of the present invention, etc., in the case of non - oral administration such as a topical skin agent, etc., it can be any itchy site without particular limitation.

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

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

[0074] <2> An agent for preventing or improving itching, wherein ozanimod, its pharmaceutically acceptable salt, or its prodrug is used as an active ingredient.

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

[0076] <4> The prophylactic or ameliorating agent for itching as described in <2>, 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, 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 prophylactic or ameliorating agent for pruritic skin diseases as described in <3>, 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.

[0078] <6> The agent as described in any one of <1> to <5>, wherein the content of ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof in the preparation is preferably 0.01% by mass or more, more preferably 0.05% by mass or more, based on the total amount of the preparation and in terms of ozanimod, and is preferably 1.2% by mass or less, more preferably 0.6% by mass or less, and is preferably 0.01% to 1.2% by mass, more preferably 0.05% to 0.6% by mass.

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

[0080] <8> The agent as described in any one of <1> to <7>, wherein when ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof is administered orally, the dosage or usage amount, in terms of ozanimod, is preferably 0.01 mg or more, more preferably 0.1 mg or more, for each adult (60 kg) per administration, and is preferably 100 mg or less, more preferably 10 mg or less, and is preferably 0.01 mg to 100 mg, more preferably 0.1 mg to 10 mg.

[0081] <9> Use of ozanimod, its pharmaceutically acceptable salts, or its prodrugs in the manufacture of a C3a receptor antagonist.

[0082] <10> An ozanimod, its pharmaceutically acceptable salts, or its prodrugs, which is used for C3a receptor antagonism.

[0083] <11> A non-therapeutic use of an ozanimod, its pharmaceutically acceptable salts, or its prodrugs, which is used for C3a receptor antagonism.

[0084] <12> A method for C3a receptor antagonism, wherein an ozanimod, its pharmaceutically acceptable salts, or its prodrugs are administered or applied to a subject.

[0085] <13> Use of ozanimod, its pharmaceutically acceptable salts, or its prodrugs in the manufacture of an agent for preventing or improving itching.

[0086] <14> An ozanimod, its pharmaceutically acceptable salts, or its prodrugs, which is used for preventing or improving itching.

[0087] <15> A non-therapeutic use of ozanimod, its pharmaceutically acceptable salts, or its prodrugs in preventing or improving itching.

[0088] <16> A method for preventing or improving itching, wherein an ozanimod, its pharmaceutically acceptable salts, or its prodrugs are administered or applied to a subject.

[0089] <17> The use as described in <13>, the ozanimod, its pharmaceutically acceptable salts, or its prodrugs 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, pruritus associated with chronic renal 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 ozanimod, its pharmaceutically acceptable salts, or its prodrugs in the manufacture of an agent for preventing or improving pruritic skin diseases.

[0091] <19> An ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof, which is used for the prevention or improvement of pruritic skin diseases.

[0092] <20> The non-therapeutic use of ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof for the prevention or improvement of pruritic skin diseases.

[0093] <21> A method for the prevention or improvement of pruritic skin diseases, wherein ozanimod, a pharmaceutically acceptable salt thereof, or a prodrug thereof is administered or applied to a subject.

[0094] <22> The use as described in <18>, the ozanimod as described in <19>, a pharmaceutically acceptable salt thereof, or a prodrug thereof, the non-therapeutic use as described in <20>, the method as described in <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, and more preferably atopic dermatitis, xeroderma or nodular prurigo.

[0095] [Examples]

[0096] Example 1 C3a receptor antagonistic effect of ozanimod

[0097] 1. C3a receptor ligand

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

[0099] 2. Evaluation samples

[0100] As a C3a receptor antagonist (antagonist), 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 ozanimod (Cayman Chemical) were used. At the following evaluation, compound 1 and ozanimod 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 an expression vector required for evaluation has been introduced according to the following is used. In a serum-free DMEM medium, a human C3a receptor expression vector (OriGene), a GNA16 expression vector, and a PEI-MAX (PolyScience) solution are sequentially added and thoroughly mixed, and then left at room temperature for 20 minutes to prepare a transfection solution. The prepared transfection solution is added to HEK293 seeded in a 6-well cell culture plate, and cultured for 24 hours or more to introduce the expression vector into HEK293. After culturing, the medium is removed, washed once with PBS, the cells are detached using 0.05% trypsin / EDTA (Gibco), and supplied to the following Ca 2+ flow detection (Ca 2+ -flux assay). Furthermore, the human GNA16 expression vector is prepared by inserting a sequence identical to the cDNA sequence of human GNA16 (Accession No. M63904.1) in the GenBank, a base sequence database provided by the National Center for Biotechnology Information, into the plasmid vector pIRESneo3 (manufactured by Clontech).

[0103] 4. Ca 2+ flow detection

[0104] The cells for evaluating the C3a receptor response are seeded in a BioCoat (registered trademark) Poly-D-Lysine 96-well Black Flat Bottom Microplate (CORNING). The intracellular Ca 2+ concentration upon C3a receptor stimulation is measured using a Fluo-4 Calcium Kit (Calcium Kit-Fluo4) (DOJINDO). A loading buffer prepared according to the kit manual is added and left at 37°C for 1 hour, whereby the Ca 2+ fluorescent indicator Fluo-4 is introduced into the cells. After washing once with HBSS, a recording buffer in which compound 1 or Ozanimod, a C3a receptor antagonist, is dissolved is added. Furthermore, a recording buffer is added to the control. After 15 minutes, a C3a receptor ligand solution (mouse TLQP-21 solution or human C3a recombinant protein solution) is added to stimulate the C3a receptor, and the Ca2+ The value obtained by subtracting the measured value (blank) obtained by adding HBSS instead of the C3a receptor ligand solution from the inflow of will be used as the measured value. Furthermore, the compound 1 solution, ozanimod, 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, 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 evaluating the C3a receptor response and the measurement of the fluorescence intensity were performed using a fluorescence microplate reader (FDSS / μCELL, Hamamatsu Photonics) equipped with an automatic dispensing function, and were carried out 3 minutes after the addition of the C3a receptor ligand solution over time. When the integrated value of the fluorescence intensity measured in the control was set to 100%, the cumulative value (%) of the fluorescence intensity of ozanimod was calculated, and the results are shown in Figure 1 .

[0105] As Figure 1 shown, the fluorescence intensity generated by the C3a receptor stimulation caused by the C3a receptor ligand measured in the control was inhibited by the addition of compound 1 or ozanimod, indicating that ozanimod has the effect of acting as 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 as animals.

[0109] Atopic dermatitis-like eczema model (AEW model): Absorbent cotton soaked in a 1:1 (v / v) mixture of acetone and diethyl ether was pressed onto the shaved skin of C57BL / 6J mice and left for 15 seconds. Immediately thereafter, 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) : Mites (M. musculi) were parasitized on NC / Nga mice to spontaneously develop dermatitis, thereby producing an AD model. As control mice, mice of the same strain raised under SPF conditions without mite parasitization were used. n = 8 for each group was used for this analysis.

[0111] 2. Purification of Total RNA

[0112] The dorsal root ganglia were removed from the cervical vertebrae of mice. The excised tissues were homogenized using a Polytron Homogenizer, and total RNA was purified using the RNeasy Mini Kit from QIAGEN.

[0113] 3. Gene expression analysis

[0114] RNA-seq : Total RNA purified from AEW model mice and their control mice was used. Reverse transcription was performed using SuperScript VILO (Thermo fisher scientific), and subsequent processing was based on the Ion AmpliSeq standard protocol of Thermo fisher scientific. Sequencing was performed using the Ion S5 system. The results are shown in Figure 2 in.

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

[0116] Real-time PCR : Total RNA purified from AEW model mice, AD model mice, and their control mice was used. Vgf was used as the target gene, and quantitative PCR analysis of the Vgf gene was performed using a real-time PCR system (Real-Time PCR System) with specific TaqMan Gene Expression Assays. Rplp0 was used as an internal standard. The relative expression levels of the Vgf gene, with the Vgf gene expression level of control mice set to 1, are shown in Figure 3 and Figure 4 in.

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

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

[0119] 1. Experimental animals

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

[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 normal saline. It was administered intradermally at 20 μL to the nape of the neck of 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 mice was observed in the form of a significant increase in Events and Beats. The manifestation of the scratching behavior is an immediate response within 30 minutes.

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

[0127] 1. Experimental animals

[0128] Mice of the WBB6F1 / Kit- Kit W / Kit W-v system were used as mast cell-deficient mice, and mice of the WBB6F1+ / + system were used as wild-type control mice. 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 normal saline. It was administered intradermally at 20 μL to the nape of the neck of 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 6In this case, the total value of consecutive scratching actions is recorded as an event.

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

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

[0135] 1. Experimental animals

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

[0137] Production 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 Co., Ltd.) 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 to 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, scratching behavior was measured for 3 hours after the final administration of the sample. MicroAct (Neuroscience Co., Ltd.) was used to measure scratching behavior. The results are shown in Figure 7 In this case.

[0142] As Figure 7 shown, in AD model mice that were administered Compound 1 which was confirmed to have a C3a receptor antagonistic effect in parallel with the application of the mite allergen, it was confirmed that scratching behavior was significantly reduced compared to AD model mice that were only administered the solvent.

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

[0144] 1. Experimental animals

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

[0146] Production of AEW model : Shave the back of the neck, press a piece of absorbent cotton soaked in a 1:1 (v / v) mixture of acetone and diethyl ether onto the back of the neck, and let it stand for 15 seconds. Immediately afterwards, press a piece of absorbent cotton soaked in water onto it for 30 seconds. Repeat this treatment twice 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 back of the neck. Only the above solvent was administered in equal amounts 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 (manufactured by Neuroscience) was used to measure scratching behavior. The results are shown in Figure 8 below.

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

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

[0153] 1. Experimental animals

[0154] Mice of the NC / Nga strain were infested with the house mouse mite (M. musculi) to spontaneously develop dermatitis, thereby establishing an AD model. Male mice at 14 weeks of age with dermatitis were used. After 1 week of acclimation, the back of the neck was shaved and the scratching behavior was measured, and they were divided into 2 groups of 8 each in such a way that the number of scratching behaviors was equal.

[0155] 2. Preparation of the evaluation sample

[0156] Use ozanimod (Selleck Biotech). Additionally, use the one prepared by making carboxymethyl cellulose (Maruishi Pharmaceutical Co., Ltd.) into 0.5% with water for injection as the solvent. The weighed ozanimod was gently crushed using an agate mortar and pestle, and then the solvent was added dropwise and mixed repeatedly to prepare a suspension (3 mg / mL). It was prepared on the day of administration and stored at room temperature until the time of administration.

[0157] 3. Administration of the evaluation sample

[0158] Measure the body weight, calculate the dosage (10 mL / kg), and orally administer the prepared evaluation sample to the mice using a mouse gavage tube and a disposable syringe.

[0159] 4. Measurement of scratching behavior

[0160] Use MicroAct (Neuroscience) to measure the scratching behavior. The number of scratching behaviors was set as the total value (events) of consecutive 1 scratching action, and the measurement time was set as 22 hours. It was measured on the day before the administration of ozanimod (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 ozanimod. The results are shown in Figure 9 in.

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

[0162] Example 3 Improvement of itching in pruritus model mice (ANIT model mice) accompanied by cholestasis caused by ozanimod

[0163] 1. Experimental animals

[0164] Use shaved male C57BL / 6J mice. α-Naphthyl isothiocyanate (ANIT) dissolved in olive oil (FUJIFILM Wako Pure Chemical Corporation) was orally administered to 8-week-old C57BL / 6J mice once a day for 10 days using a mouse gavage tube and a disposable syringe. When administering, the body weight was measured in advance, and the dosage was adjusted to 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 the evaluation sample

[0166] Ozanimod (Selleck Biotech) was used. Additionally, a solvent prepared by making carboxymethyl cellulose (Maruishi Pharmaceutical Co., Ltd.) into 0.5% with water for injection was used. The weighed ozanimod was gently crushed using an agate mortar and pestle, and then the solvent was added dropwise and mixed repeatedly to prepare a suspension (3 mg / mL). It was prepared on the day of administration and stored at room temperature until administration.

[0167] 3. Administration of the evaluation sample

[0168] The body weight was measured, and the dosage (10 mL / kg) was calculated. The prepared evaluation sample was administered orally using a mouse gavage tube and a disposable syringe.

[0169] 4. Measurement of scratching behavior

[0170] MicroAct (Neuroscience) was used to measure the scratching behavior. The number of scratching behaviors was set as the total value (events) of consecutive 1 scratching action, and the measurement time was set as 22 hours. It was measured on the day before administration of ozanimod (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 ozanimod. The results are shown in Figure 10 below.

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

Claims

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

2. An agent for preventing or improving itching, wherein ozanimod, 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 xeroderma, itching in prurigo nodularis, or pruritus associated with cholestasis.

5. An agent for preventing or improving pruritic skin diseases, wherein ozanimod, 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, xeroderma or prurigo nodularis.

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

8. Use of ozanimod, 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 xeroderma, itching in prurigo nodularis, or pruritus associated with cholestasis.

11. Use of ozanimod, 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, xeroderma or prurigo nodularis.

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

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

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

16. The ozanimod, 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 xeroderma, itching in prurigo nodularis, or pruritus associated with cholestasis.

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

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

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

20. Non-therapeutic use of ozanimod, 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 ozanimod, 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 ozanimod, its pharmaceutically acceptable salt, or its prodrug is administered or applied to a subject.

26. A method for preventing or improving itching, wherein ozanimod, 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 ozanimod, 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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