Treatment for eosinophilic granulomatosis with polyangiitis

Abstract

To provide a novel agent for treating eosinophilic granulomatosis with polyangiitis (EGPA). [Solution] A treatment agent for EGPA comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate, its pharmaceutically acceptable salt, or its hydrate.

Description

[Technical Field]

[0001] The present invention relates to a therapeutic agent for eosinophilic granulomatosis with polyangiitis, etc. [Background technology]

[0002] Eosinophilic granulomatosis with polyangiitis (EGPA) is a rare, intractable disease of unknown etiology. It is preceded by asthma and allergic rhinitis, both of which are characterized by a marked increase in eosinophils, several years before the onset of the disease. This is followed by a marked increase in peripheral blood eosinophils, which leads to eosinophilic inflammation in various organs throughout the body and granulomatous necrotizing vasculitis of small to medium-sized blood vessels. Symptoms associated with vasculitis include fever, joint pain, muscle pain, and weight loss, with heart, gastrointestinal, and neurological damage particularly common. Skin symptoms such as purpura and kidney damage also occur. Severe damage to the heart and gastrointestinal tract can be fatal, so prompt treatment is essential.

[0003] Drug therapy for EGPA involves administering high doses of glucocorticoids alone or in combination with immunosuppressants, etc., to induce remission, and then gradually reduce the dose of glucocorticoids to maintain remission (Non-patent Document 1, Non-patent Document 2).

[0004] In recent years, it has been confirmed that the anti-IL-5 antibody mepolizumab (NUCALA (registered trademark), GlaxoSmithKline) and the anti-IL-5 receptor antibody benralizumab (FASENRA (registered trademark), AstraZeneca) have the effect of suppressing EGPA relapse and reducing glucocorticoid dosage, and these have been approved as drugs for the treatment of EGPA (Non-patent Document 3, Non-patent Document 4).

[0005] However, it has been shown that patients without high eosinophil counts have limited response to mepolizumab (Non-Patent Document 3). Because pathways other than eosinophilic inflammation and various cytokines are thought to be involved in the pathogenesis of EGPA, more optimal therapeutic approaches that can overcome this problem are still needed (Non-Patent Document 5).

[0006] On the other hand, the following structural formula: [ka] Methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate, represented by the formula (I), is a Janus kinase (JAK) 1 inhibitor involved in the transmission of various cytokine signals. Therefore, this compound is expected to be applicable to diseases involving JAK1, such as inflammatory diseases, autoimmune diseases, and proliferative diseases (Patent Document 1). However, it has not been reported that this compound improves vascular lesions associated with EGPA, and therefore can be used to treat EGPA. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2017 / 150477 [Patent Document 2] International Publication No. 2016 / 035814 [Non-patent literature]

[0008] [Non-Patent Document 1] Chung et al., Arthritis & Rheumatology Vol. 73, No. 8, August 2021, pp 1366-1383 [Non-patent document 2] Hellmich B et al., Ann Rheum Dis 2024;83:30-47 [Non-patent document 3] Wechsler et al., N Engl J Med 2017;376:1921-32 [Non-patent document 4] Wechsler et al., N Engl J Med 2024;390:911-21 [Non-patent document 5] Raffray and Guillevin, Presse Med 49 (2020) 104036 [Non-patent document 6] Yamauchi et al., Experimental Lung Research, 36, 227-236, 2010 Summary of the Invention [Problem to be solved by the invention]

[0009] The problem to be solved by the present invention is to provide a novel agent for treating EGPA, in particular an agent for treating EGPA that is highly effective against EGPA and has few side effects. [Means for solving the problem]

[0010] The present inventors have discovered a compound having the following structural formula: [ka] The present inventors have found that methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate (hereinafter referred to as "compound (1)") represented by the formula (I), a pharmaceutically acceptable salt thereof, or a hydrate thereof can suppress vascular lesions associated with EGPA in mice with few side effects, and have thus completed the present invention.

[0011] That is, the present invention includes the following aspects: [1] A therapeutic agent for eosinophilic granulomatosis with polyangiitis (EGPA), comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate, a pharmaceutically acceptable salt thereof, or a hydrate thereof. [2] A treatment for EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate. [3] A treatment for EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate. [4] The agent for treating EGPA according to any one of [1] to [3] above, wherein the treatment of EGPA includes suppression of vascular lesions associated with EGPA. [5] A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and a pharmaceutically acceptable carrier. [6] A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate. [7] A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate. [8] The pharmaceutical composition for treating EGPA according to any one of [5] to [7] above, wherein the treatment of EGPA includes suppression of vascular lesions associated with EGPA. [Effects of the Invention]

[0012] According to the present invention, it is possible to provide a treatment agent for EGPA that has few side effects and can be used safely, more specifically, a treatment agent for EGPA that has little effect on platelets, red blood cells, and body weight and can significantly suppress vascular lesions associated with EGPA. [Brief explanation of the drawings]

[0013] [Figure 1] 1 shows a powder X-ray diffraction spectrum chart of a monohydrate crystal of the tosylate salt of Compound (1). The vertical axis represents peak intensity (cps) and the horizontal axis represents diffraction angle (2θ [°]). [Figure 2] Representative photographs of hematoxylin-eosin stained specimens showing each vascular lesion grade are shown. The scale bar indicates 50 μm. [Figure 3] Ovalbumin (OVA)-induced eosinophilic vasculitis mouse model was administered 50 and 200 mg / kg of the monohydrate tosylate of compound (1) (Examples 1 and 2), 50 and 150 mg / kg of tofacitinib citrate (Comparative Examples 1 and 2), and 15 and 45 mg / kg of prednisolone (Comparative Examples 3 and 4), and the results of histopathological examination are shown. [Figure 4] 1 shows the results of evaluating changes in blood platelets when the monohydrate tosylate of compound (1), tofacitinib citrate, and prednisolone were administered to a mouse model of OVA-induced eosinophilic vasculitis. [Figure 5] 1 shows the results of evaluating changes in the number of red blood cells in the blood when the monohydrate tosylate of compound (1), tofacitinib citrate, and prednisolone were administered to a mouse model of OVA-induced eosinophilic vasculitis. [Figure 6]1 shows the results of evaluating changes in body weight when the monohydrate tosylate of Compound (1), tofacitinib citrate, and prednisolone were administered to a mouse model of OVA-induced eosinophilic vasculitis. DETAILED DESCRIPTION OF THE INVENTION

[0014] Compound (1) herein, a pharmaceutically acceptable salt thereof, or a hydrate thereof can be prepared according to the method described in Patent Document 1 or Patent Document 2, for example.

[0015] As used herein, pharmaceutically acceptable salts of compound (1) include, for example, salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc., and salts of organic acids such as acetic acid, citric acid, tartaric acid, maleic acid, succinic acid, fumaric acid, p-toluenesulfonic acid (tosylate), benzenesulfonic acid, methanesulfonic acid, etc. Preferably, p-toluenesulfonic acid (tosylate) salt of compound (1) is used. As used herein, the hydrate of compound (1) or a pharmaceutically acceptable salt thereof may include, for example, a monohydrate or a dihydrate. When a hydrate is formed, it may be coordinated with any type and number of water molecules. A preferred example is the p-toluenesulfonic acid (tosylic acid) monohydrate of compound (1).

[0016] In the present invention, compound (1), its pharmaceutically acceptable salt, or its hydrate may be crystalline or amorphous. Preferred examples include a crystalline form, such as a crystalline monohydrate of the tosylate salt of compound (1), which exhibits diffraction peaks at the following diffraction angles 2θ: 12.6 degrees, 13.3 degrees, 17.2 degrees, 20.6 degrees, and 21.8 degrees in a powder X-ray diffraction spectrum obtained using Cu Kα radiation. An example of a powder X-ray diffraction spectrum chart of the crystal of the above form is shown in Figure 1. The powder X-ray diffraction spectrum chart was measured using SmartLab (manufactured by Rigaku Corporation) (target: Cu, voltage: 45 kV, current: 200 mA, scan speed: 47.3 degrees / min).

[0017] Compound (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof can inhibit vascular lesions associated with EGPA. Thus, one embodiment of the present invention includes a method for treating EGPA, comprising administering compound (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein, in some embodiments, the treatment of EGPA includes inhibiting vascular lesions associated with EGPA.

[0018] As used herein, "treatment" of EGPA refers to any treatment administered to a patient suffering from EGPA, including, for example, preventing, suppressing, delaying, or halting the onset of one or more symptoms of EGPA (e.g., vascular lesions), alleviating, mitigating, ameliorating, or eliminating symptoms, suppressing, delaying, or halting the progression or worsening of symptoms, etc. Treatment may be administered before the onset of symptoms as a preventative measure, or alternatively, treatment may be administered after the onset of the disease.

[0019] The agent for treating EGPA according to the present invention can contain compound (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof, either as is or in a pharmaceutically acceptable carrier, in an amount of, for example, 0.01 to 99.5%, preferably 0.5 to 90%. Accordingly, one embodiment of the present invention includes a pharmaceutical composition for treating EGPA, comprising Compound (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof, and a pharmaceutically acceptable carrier, wherein, in some embodiments, treating EGPA includes inhibiting vascular lesions associated with EGPA.

[0020] As used herein, the term "pharmaceutically acceptable carrier" refers to various carriers commonly used in the field of formulation, such as solid, semi-solid, or liquid diluents, fillers, and other formulation auxiliaries. One or more of these may be used.

[0021] The agent or pharmaceutical composition for treating EGPA or vascular lesions associated with EGPA according to the present invention (hereinafter collectively referred to as "agent") can take the form of any of oral preparations such as powders, capsules, tablets, sugar-coated preparations, granules, powders, suspensions, liquids, syrups, elixirs, and lozenges, or parenteral preparations such as injections and suppositories, in solid or liquid dosage units. It may also be a sustained-release preparation. Of these, oral preparations such as tablets are particularly preferred.

[0022] The powdered formulation can be produced by crushing compound (1), a pharmaceutically acceptable salt thereof, or a hydrate thereof (hereinafter collectively referred to as "compound (1), etc.") into an appropriate fine powder by a method commonly used in the pharmaceutical field.

[0023] Powders can be prepared by crushing Compound (1) or the like into suitable fine particles using a method commonly used in the pharmaceutical field, and then mixing the resulting mixture with a similarly crushed pharmaceutically acceptable carrier, such as an edible carbohydrate such as starch or mannitol. Optionally, flavoring agents, preservatives, dispersing agents, coloring agents, fragrances, etc. can be added.

[0024] Capsules can be prepared by first filling the powdered or powdered formulations, or the granulated formulations described above, into capsule shells, such as gelatin capsules, using methods commonly used in the pharmaceutical field. Capsules can also be prepared by mixing a lubricant or a flow agent, such as colloidal silica, talc, magnesium stearate, calcium stearate, or solid polyethylene glycol, with the powdered or powdered formulations and then filling the capsules. The addition of a disintegrant or solubilizer, such as carboxymethylcellulose, carboxymethylcellulose calcium, low-substituted hydroxypropylcellulose, croscarmellose sodium, carboxymethyl starch sodium, calcium carbonate, or sodium carbonate, can improve the efficacy of the drug when the capsule is ingested. Alternatively, a fine powder of Compound (1) or the like can be suspended in vegetable oil, polyethylene glycol, glycerin, or a surfactant, and then enclosed in a gelatin sheet to form a soft capsule.

[0025] Tablets can be produced by adding an excipient to powdered compound (1) or the like to prepare a powder mixture, granulating or slugging the mixture, adding a disintegrant or lubricant, and then compressing the mixture into tablets, according to a method commonly used in the pharmaceutical field.

[0026] The powder mixture can be prepared by mixing appropriately powdered Compound (1) or the like with a diluent or base by a method commonly used in the pharmaceutical field. If necessary, binders (e.g., sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, gelatin, polyvinylpyrrolidone, polyvinyl alcohol), dissolution retardants (e.g., paraffin), resorbents (e.g., quaternary salts), adsorbents (e.g., bentonite, kaolin), etc. can be added.

[0027] The powder mixture can be first moistened with a binder, for example, syrup, starch paste, gum arabic, cellulose solution, or polymer solution, and then mixed by stirring, dried, and milled to form granules, as is conventionally done in the pharmaceutical arts. Instead of granulating the powder in this way, it is also possible to first run the mixture through a tablet press, and then break up the resulting imperfectly formed slugs into granules. The granules thus produced can be lubricated with the addition of stearic acid, a stearate salt, talc, mineral oil, or the like to prevent them from sticking together.

[0028] Alternatively, tablets can be produced by mixing compound (1) or the like with a fluid inert carrier and then directly compressing the mixture into tablets, without going through the granulation or slugging steps as described above.

[0029] The tablets thus produced can be film-coated or sugar-coated by a method commonly used in the pharmaceutical field. Transparent or semi-transparent protective coatings such as a sealing coating of shellac, coatings of sugars or polymeric materials, and polishing coatings made of wax can also be used.

[0030] Other oral administration preparations, such as liquids, syrups, troches, and elixirs, can be prepared in dosage unit forms so that a given amount thereof contains a given amount of Compound (1), etc.

[0031] Syrups can be prepared by dissolving Compound (1) or the like in a suitable flavored aqueous solution by a method commonly used in the pharmaceutical field, and elixirs can be prepared by using a pharmaceutically acceptable alcoholic carrier.

[0032] Suspensions can be prepared by dispersing compound (1) or the like in a pharmaceutically acceptable carrier using a method commonly used in the pharmaceutical field. If necessary, solubilizers, emulsifiers (e.g., ethoxylated isostearyl alcohols, polyoxyethylene sorbitol esters), preservatives, flavoring agents (e.g., peppermint oil, saccharin), etc. can be added.

[0033] If necessary, the dosage unit formulation for oral administration can be microencapsulated by methods commonly used in pharmaceutical formulations. The formulation can also be coated or embedded in polymers, waxes, etc. to provide extended action or sustained release.

[0034] Parenteral preparations can be in the form of liquid dosage units, such as solutions or suspensions, for subcutaneous, intramuscular, or intravenous injection. These parenteral preparations can be prepared by suspending or dissolving a predetermined amount of compound (1) or the like in a pharmaceutically acceptable liquid carrier suitable for the purpose of injection, such as an aqueous or oily medium, and then sterilizing the suspension or solution, using methods commonly used in the pharmaceutical field. Pharmaceutically acceptable salts or salt solutions can be added to make the injection isotonic. Stabilizers, preservatives, emulsifiers, etc. can also be added.

[0035] Suppositories can be prepared by dissolving or suspending compound (1) or the like in a low-melting, water-soluble or insoluble solid, such as polyethylene glycol, cocoa butter, semi-synthetic fats and oils (e.g., Witepsol (registered trademark)), higher esters (e.g., myristyl palmitate), or a mixture thereof.

[0036] The dosage of the agent for treating EGPA according to the present invention varies depending on the patient's condition, such as body weight and age, the route of administration, the severity of symptoms, etc., but generally, for an adult, the appropriate amount is 0.001 mg to 100 mg per day of compound (1) or the like. In some cases, a lower dose may be sufficient, while in other cases a higher dose may be required. The agent can be administered once or several times a day, or at intervals of one day to several days. [Example]

[0037] The present invention will be explained in more detail below with reference to Examples, Reference Examples, Comparative Examples and Test Examples, but the present invention is not limited to the scope shown below.

[0038] Reference Example 1 3-amino-4-hydroxy-1,3-thiazolidine-2-thione [ka] Sodium borohydride (19.1 g) was added to tetrahydrofuran (750 mL), and a slurry of N-aminorhodanine (250 g) in tetrahydrofuran (500 mL) was added in portions at 5°C or below. After stirring for 30 minutes at 5°C or below, methanol (111 mL) was added dropwise and the mixture was stirred for 2 hours. Concentrated hydrochloric acid (44 mL) diluted with water (500 mL) was added dropwise, followed by the dropwise addition of water (1000 mL), and the mixture was stirred for 1 hour at 10°C or below. The precipitated crystals were filtered, washed with water (600 mL), and dried under reduced pressure at 40°C to obtain the title compound (209.1 g). MS(m / z): 151[M+H] +

[0039] Reference Example 2 Sodium 2-chloro-2-cyanoethene-1-oleate [ka] To a slurry of sodium methoxide (14.3 g) in cyclopentyl methyl ether (300 mL), methyl formate (17.5 g) was added dropwise at 20° C. or below, followed by dropwise addition of chloroacetonitrile (20 g), and the mixture was stirred for 3 hours at 30° C. or below. After completion of the reaction, the precipitated crystals were filtered, washed with cyclopentyl methyl ether (40 mL), and then dried under reduced pressure at 40° C. to obtain the title compound (29.2 g).

[0040] Reference Example 3 Methyl pyrazolo[5,1-b][1,3]thiazole-7-carboxyimidate hydrochloride [ka] Concentrated sulfuric acid (48.97 g) was added to a slurry of 3-amino-4-hydroxy-1,3-thiazolidine-2-thione (50 g) obtained in Reference Example 1 in 2-propanol (250 mL), and the mixture was heated and stirred at 80°C for 1 hour. After cooling, acetonitrile (500 mL) and sodium 2-chloro-2-cyanoethene-1-olate (62.65 g) obtained in Reference Example 2 were added at 40°C or below, and the mixture was stirred at 80°C for 4 hours. After cooling, activated carbon (10 g) was added, and the mixture was stirred at room temperature for 30 minutes. The insoluble matter was filtered and washed three times with acetonitrile (100 mL). The filtrate was concentrated under reduced pressure, followed by azeotropic distillation three times with methanol (100 mL) to remove the acetonitrile. To the concentrate, methanol (150 mL) and tetrahydrofuran (150 mL) were added. This mixture was added to a solution prepared from methanol (350 mL) and acetyl chloride (209 g) at 20°C or below. After stirring overnight at room temperature, tetrahydrofuran (350 mL) was added and the mixture was further stirred for 1 hour at below 10° C. The precipitated crystals were filtered, washed with tetrahydrofuran (300 mL), and then dried under reduced pressure at 50° C. to obtain the title compound (45.5 g). MS(m / z):182[M+H] +

[0041] Reference Example 4 Pyrazolo[5,1-b][1,3]thiazole-7-carboximidamide acetate [ka] To a slurry of methyl pyrazolo[5,1-b][1,3]thiazole-7-carboxyimidate hydrochloride (200 g) obtained in Reference Example 3 in methanol (1000 mL) was added ammonium acetate (85.15 g), followed by DIPEA (142.82 g), and the mixture was stirred at 65°C for 1 hour. After the reaction was completed, the mixture was cooled, and acetonitrile (2000 mL) was added dropwise at room temperature, followed by stirring at 10°C or below for 1 hour. The precipitated crystals were filtered, washed with acetonitrile (400 mL), and then dried under reduced pressure at 50°C to obtain the title compound (185.12 g). Elemental analysis (as C6H6N4S·C2H4O2) Calculated values ​​(%) C: 42.47, H: 4.46, N: 24.76 Actual measured values ​​(%) C: 42.18, H: 4.25, N: 24.41

[0042] Reference Example 5 6-Hydroxy-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidine-4-carboxylic acid [ka] Sodium diethyl oxalacetate (130.65 g) was added to an aqueous solution (900 mL) of sodium hydroxide (39.08 g) at 10°C or below, and the mixture was stirred for 1 hour. To this solution, pyrazolo[5,1-b][1,3]thiazole-7-carboximidamide acetate (90 g) obtained in Reference Example 4 was added, and the mixture was heated and stirred at 50°C for 3 hours. The mixture was then cooled, and concentrated hydrochloric acid (138 g) was added at 30°C or below. After confirming that the pH of the mixture had reached 1 to 2, the mixture was stirred overnight at room temperature. The precipitated crystals were collected by filtration, washed with water (360 mL), and dried at 60°C to obtain the title compound (107.17 g). MS(m / z):263[M+H] +

[0043] Reference Example 6 Methyl {1-[6-hydroxy-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidine-4-carbonyl]piperidin-4-yl}carbamate [ka] Triethylamine (275.51 g) was added to a slurry of 6-hydroxy-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidine-4-carboxylic acid (238 g) obtained in Reference Example 5 in DMF (714 mL) and the mixture was stirred at 50°C for 30 minutes. After cooling, 1,1'-carbonyldiimidazole (323.76 g) was added at 20°C or below. After stirring for 30 minutes, methyl piperidin-4-ylcarbamate tosylate (449.79 g) was added and the mixture was stirred for 30 minutes. After completion of the reaction, acetonitrile (3570 mL) was added dropwise at room temperature and the mixture was stirred overnight. The precipitated crystals were filtered, washed with acetonitrile (480 mL), and then dried under reduced pressure at 60°C to obtain the title compound (377.79 g). MS(m / z):403[M+H] +

[0044] Reference Example 7 6-{4-[(methoxycarbonyl)amino]piperidine-1-carbonyl}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl 4-methylbenzene-1-sulfonate [ka] Triethylamine (275.33 g) was added to a slurry of methyl {1-[6-hydroxy-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidine-4-carbonyl]piperidin-4-yl}carbamate (365 g) obtained in Reference Example 6 in acetonitrile (1825 mL), followed by the addition of 4-toluenesulfonyl chloride (259.36 g), and the mixture was heated and stirred at 50 ° C. for 1 hour. After the reaction was completed, the mixture was cooled, and water (3650 mL) was added dropwise at room temperature, followed by stirring at 10 ° C. or below for 1 hour. The precipitated crystals were filtered, washed with water (730 mL), and then dried under reduced pressure at 60 ° C. to obtain the title compound (442.08 g). MS(m / z):557[M+H] +

[0045] Reference Example 8 Methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate [Step 1] Preparation of methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate (Compound 1) To a solution of 6-{4-[(methoxycarbonyl)amino]piperidine-1-carbonyl}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl 4-methylbenzene-1-sulfonate (1.0 g) obtained in Reference Example 7 in acetonitrile (7.0 mL), DIPEA (0.69 g) and (1S)-1-cyclopropylethanamine (460 mg) were added, the tube was sealed, and the mixture was heated and stirred at 100°C for 2 hours. After completion of the reaction, the reaction solution was diluted with ethyl acetate and washed with water (50 mL) and saturated brine (30 mL). The organic layer was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, and then purified by column chromatography to give the title compound (720 mg). [Step 2] Methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate Acetonitrile (5 mL) was added to methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate (201 mg) obtained in Step 1, and the mixture was heated to 50°C. p-Toluenesulfonic acid monohydrate (81 mg) was added, and the mixture was stirred at room temperature overnight. The precipitated crystals were collected by filtration and dried under reduced pressure to obtain crystals of the title compound (237 mg). Elemental analysis of the obtained crystals showed that they were monohydrate. Elemental analysis value (C 22 H 27 (as N7O3S·C7H8O3S·1.0H2O) Calculated values ​​(%) C: 52.79, H: 5.65, N: 14.86 Actual measured values ​​(%) C: 52.72, H: 5.54, N: 14.82 Specific rotation [α] D 25 =-44.4 (c=1.00, DMSO) The powder X-ray diffraction spectrum of the obtained crystals was measured using a SmartLab (Rigaku Corporation) (target: Cu, voltage: 45 kV, current: 200 mA, scan speed: 47.3 degrees / min). The results are shown in Figure 1. As can be seen from Figure 1, the crystals exhibited diffraction peaks at the following diffraction angles 2θ: 12.6 degrees, 13.3 degrees, 17.2 degrees, 20.6 degrees, and 21.8 degrees.

[0046] Test Example 1: Vascular lesion suppression effect in a mouse model of ovalbumin (OVA)-induced eosinophilic vasculitis To evaluate the efficacy and safety of compound (1) in EGPA, we investigated the OVA-induced eosinophilic vasculitis in a mouse model using the pan-JAK inhibitor tofacitinib citrate and prednisolone as controls.

[0047] (1) Preparation of the medium Methylcellulose (Metolose (registered trademark), SM-400, Shin-Etsu Chemical Co., Ltd.) was dissolved in distilled water to prepare a 0.5 w / v % aqueous methylcellulose solution.

[0048] (2) Preparation method of the drug solution (a) Monohydrate of the tosylate salt of compound (1) (Examples 1 and 2) A 5 mg / mL suspension (Example 1) or a 20 mg / mL suspension (Example 2) of Compound (1) in a 0.5 w / v% aqueous methylcellulose solution was prepared by adding a 0.5 w / v% aqueous methylcellulose solution to the monohydrate of tosylate of Compound (1) obtained in Step 2 of Reference Example 8. (b) Tofacitinib Citrate (Comparative Examples 1 and 2) Tofacitinib citrate was obtained from Lanzhou Boc Chemical (Shanghai, China). A 5 mg / mL suspension (Comparative Example 1) or a 15 mg / mL suspension (Comparative Example 2) of tofacitinib citrate in a 0.5 w / v% aqueous methylcellulose solution was prepared by adding a 0.5 w / v% aqueous methylcellulose solution to tofacitinib citrate. (c) Prednisolone (Comparative Examples 3 and 4) Prednisolone was obtained from Nacalai Tesque (Kyoto, Japan). A 0.5 w / v % aqueous solution of methylcellulose was added to prednisolone to prepare a 1.5 mg / mL prednisolone suspension (Comparative Example 3) or a 4.5 mg / mL prednisolone suspension (Comparative Example 4).

[0049] (3) Test method We optimized the OVA-induced eosinophilic vasculitis mouse model reported by Yamauchi et al. (Non-Patent Document 6). The day of the first immunization was defined as day 0. OVA sensitization solution (0.5 mL) was administered intraperitoneally to female C57BL / 6NCrSlc mice (6 weeks old) on days 0 and 5. OVA sensitization solution was prepared by adsorbing OVA (8 μg / mouse; Sigma-Aldrich, Steinheim, Germany) onto 20% aluminum gel (Imject® Alum, Thermo Fisher Scientific, Sunnyvale, CA, USA) in phosphate-buffered saline (PBS). All mice, including those in the saline inhalation group (saline / vehicle group), were immunized. To confirm the OVA sensitization status, 50 μL of blood was collected from each mouse's tail on day 12, and plasma was obtained. Plasma anti-OVA-IgE levels were measured using an ELISA kit (DS Mouse IgE ELISA (OVA), DS Pharma Biomedical, Osaka). On day 14, the top 80 mice with the highest anti-OVA-IgE levels were selected and divided equally into eight groups (10 mice per group). From days 17 to 23, 0.5 w / v% methylcellulose aqueous solution (vehicle) or drug was orally administered at 10 mL / kg twice daily (6 h intervals). Body weight was measured daily from days 17 to 23. After the first administration of each day, mice were placed in a plastic chamber (23 × 23 × 27 cm, custom-made, Natsume Seisakusho Co., Ltd., Tokyo) and exposed to aerosolized 1% w / v OVA in saline using a nebulizer (PARI BOY SX, Pari GmbH, Starnberg, Germany) for 1 hour per day for 7 consecutive days. The saline inhalation group (saline / vehicle group) was exposed to aerosolized saline.

[0050] On day 24, mice were anesthetized with isoflurane and blood was collected via the heart. Blood samples were analyzed using a hematology testing system (ADVIA 2120i; Siemens Healthineers, Tokyo) to measure lymphocyte, eosinophil, platelet, and red blood cell counts. The bronchioles leading to the left upper lobe of the lung were tied with a string, and a tube was inserted into the trachea. The lungs were washed with a buffer solution (0.1% bovine serum albumin (w / v) and 50 μM ethylenediaminetetraacetic acid-2Na in PBS(-)). The left upper lobe of the lung was fixed in 10% neutral-buffered formalin. Hematoxylin-eosin-stained specimens were analyzed to evaluate pathological changes in the pulmonary arteries. Ten arteries (40–70 μm in diameter) were analyzed and graded on a scale of 0 to 4 according to a previous study by Yamauchi et al. Grade 0: no abnormalities; Grade 1: Minimal (no abnormalities in the arteries, very mild inflammatory cell infiltration around the arteries); Grade 2: mild (no abnormalities in the arteries, inflammatory cell infiltration around the arteries); Grade 3: Moderate (arterial wall thickening regardless of the degree of inflammatory cell infiltration); Grade 4: Severe (there are tissue changes indicating arterial damage, such as destruction of arterial elastic fibers and proliferation of mesenchymal cells into the arterial lumen). The average grade of the 10 arteries was calculated and used as the grade for each mouse (Figure 2).

[0051] (4) Results The effects of the compounds on pulmonary arterial vasculitis were examined histopathologically. Vasculitis formation, as expressed by vascular lesion grade, was significantly induced in the OVA / vehicle group compared to the saline / vehicle group. Administration of compound (1) tosylate monohydrate at 50 and 200 mg / kg (Examples 1 and 2), tofacitinib citrate at 50 and 150 mg / kg (Comparative Examples 1 and 2), or prednisolone at 15 and 45 mg / kg (Comparative Examples 3 and 4) significantly suppressed the vascular lesion grade compared to the OVA / vehicle group (Figure 3). Additionally, changes in blood platelet and red blood cell counts and body weight were evaluated as indicators of side effects. At the same doses that demonstrated efficacy, compound (1) tosylate monohydrate had no effect on platelets, red blood cells, or body weight (Figures 4, 5, and 6). In contrast, tofacitinib citrate significantly increased platelet count and body weight (Figures 4 and 6). Prednisolone significantly increased red blood cell count and decreased body weight (Figures 5 and 6).

Claims

1. A therapeutic agent for eosinophilic granulomatosis with polyangiitis (EGPA), comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

2. A treatment for EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate.

3. A treatment for EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate.

4. The agent for treating EGPA according to any one of claims 1 to 3, wherein the treatment of EGPA includes suppression of vascular lesions associated with EGPA.

5. A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate, a pharmaceutically acceptable salt thereof, or a hydrate thereof, and a pharmaceutically acceptable carrier.

6. A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate.

7. A pharmaceutical composition for treating EGPA, comprising methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate tosylate monohydrate.

8. The pharmaceutical composition for treating EGPA according to any one of claims 5 to 7, wherein the treatment of EGPA includes suppressing vascular lesions associated with EGPA.

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