Crystal of pyrrolidine compound

Abstract

The present invention relates to a crystal of a pyrrolidine compound. Provided is a crystal of 1-{2-[(3S, 4R)-1-[(3R, 4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl) pyrrolidine-3-carbonyl]-4-(methoxymethyl) pyrrolidine-3-yl]-5-(trifluoromethyl) phenyl} piperidine-4-carboxylic acid, which has a certain quality and can be used as a raw material for a pharmaceutical product. Specifically, the present invention provides a crystal comprising an equal molar amount of 1-{2-[(3S, 4R)-1-[(3R, 4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl) pyrrolidin-3-carbonyl]-4-(methoxymethyl) pyrrolidin-3-yl]-5-(trifluoromethyl) phenyl} piperidine-4-carboxylic acid and phosphoric acid.

Description

[0001] This application is a divisional application of a patent application for an invention titled "Crystal of Pyrrolidine Compound" with an application date of December 27, 2019, an application number of 201980086473.9 (PCT / JP2019 / 051570). Technical Field

[0002] The present invention relates to a crystal comprising 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid (hereinafter, sometimes referred to as "pyrrolidine compound A" or "compound A"). More specifically, it relates to a crystal comprising equimolar amounts of pyrrolidine compound A and phosphoric acid (hereinafter, sometimes referred to as "the crystal of the present invention"), which has excellent properties as a pharmaceutical raw material, and a pharmaceutical composition containing the same as an active ingredient. Background Art

[0003] Pamphlet of International Publication WO2015 / 182723 (hereinafter referred to as Patent Document 1) discloses a pyrrolidine compound having melanocortin receptor 1 (MC1R) agonistic activity (agonist activity) or a pharmaceutically acceptable salt thereof, and these compounds and a pharmaceutical composition containing these compounds as an active ingredient are useful in the treatment or prevention of various diseases that are expected to improve the condition through the activation of MC1R. Example 19 describes the hydrochloride salt of pyrrolidine compound A. However, Patent Document 1 does not disclose or teach the crystal of pyrrolidine compound A.

[0004] Prior Art Documents

[0005] Patent Documents

[0006] Patent Document 1: Pamphlet of International Publication WO2015 / 182723 Summary of the Invention

[0007] Problems to be Solved by the Invention

[0008] The problem of the present invention is to provide a crystal of pyrrolidine compound A having a certain quality that can be used as a pharmaceutical raw material.

[0009] Means for Solving the Problems

[0010] It was found that the hydrochloride salt of pyrrolidine compound A does not crystallize, is deliquescent, and is not suitable as a pharmaceutical active ingredient. Therefore, for pyrrolidine compound A, in order to obtain crystals with a certain quality suitable for use as a pharmaceutical active ingredient, the inventors of the present application tried crystallization under more than 1000 conditions. As a result, it was found that, considering purity, thermal stability, hygroscopicity, deliquescence, chemical stability, and safety, crystals containing equimolar amounts of pyrrolidine compound A and phosphoric acid are crystals with a certain quality suitable for use as a pharmaceutical active ingredient, thus completing the present invention.

[0011] In addition, it was found that it is difficult to crystallize the crystals containing equimolar amounts of pyrrolidine compound A and phosphoric acid. Therefore, the inventors of the present application studied conditions with good reproducibility that can obtain crystals with sufficient purity in a short time. It was confirmed that depending on the crystallization temperature and the composition of the crystallization solvent, there are problems such as an increase in impurities, a delay in precipitation, and deterioration of stirring fluidity and filterability due to the refinement of the crystals. To solve these problems, the inventors of the present application not only studied in depth the types or amounts, ratios, etc. of the reagents and solvents used in crystallization, but also studied in depth the crystallization steps, etc., and found a method for efficiently obtaining crystals with few impurities, good operability such as filtration, and having a quality suitable for a pharmaceutical active ingredient.

[0012] The present invention relates to the following. [1]

[0014] Crystals comprising equimolar amounts of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidin-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid. [2]

[0016] The crystals according to [1], which are formed from equimolar amounts of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidin-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid. [3]

[0018] The crystals according to any one of [1] or [2], which are co-crystals of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidin-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid. [4]

[0020] The crystal according to any one of [1] to [3], wherein in the powder X-ray diffraction spectrum, peaks are shown at diffraction angles represented by 2θ of 5.7°, 11.5°, 13.9°, 19.0° and 21.9° (±0.2° respectively). [5]

[0022] The crystal according to any one of [1] to [4], wherein in the differential scanning calorimetry analysis, an endothermic peak is present at 230 °C to 240 °C. [6]

[0024] The crystal according to any one of [1] to [5], which is obtained by adding a seed crystal to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidine-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid and a good solvent. [7]

[0026] The crystal according to any one of [1] to [6], which is obtained by the following method: adding a poor solvent to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidine-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid and a good solvent, adding a seed crystal, and then further adding a poor solvent. [8]

[0028] A melanocortin receptor 1 agonist, which contains the crystal according to any one of [1] to [7] as an active ingredient. [9]

[0030] A pharmaceutical composition, which contains the crystal according to any one of [1] to [7] and a pharmaceutically acceptable additive.

[10]

[0032] The pharmaceutical composition according to [9], which is used for preventing or treating a disease whose condition is expected to be improved by activation of the melanocortin receptor 1.

[11]

[0034] The pharmaceutical composition according to

[10] , wherein the disease is one or more diseases selected from rheumatoid arthritis, gouty arthritis, osteoarthrosis, inflammatory bowel disease, systemic scleroderma, psoriasis, fibrosis, protoporphyria, systemic lupus erythematosus, melanoma, skin cancer, vitiligo, alopecia, pain, ischemia / reperfusion injury, inflammatory diseases of the brain, hepatitis, sepsis / septic shock, nephritis, transplantation, exacerbation of HIV disease, vasculitis, uveitis, retinitis pigmentosa, age-related macular degeneration, microbial infection, celiac disease, nephrotic syndrome, and melanoma infiltration.

[12]

[0036] A method for preventing or treating a disease whose condition is expected to be improved by activation of the melanocortin receptor 1, which comprises administering to a patient an effective amount of the crystal according to any one of [1] to [7].

[13]

[0038] Use of the crystal according to any one of [1] to [7] in the manufacture of a medicament for preventing or treating a disease whose condition is expected to be improved by activation of the melanocortin receptor 1.

[14]

[0040] The crystal according to any one of [1] to [7], which is used for preventing or treating a disease whose condition is expected to be improved by activation of the melanocortin receptor 1.

[0041] The present invention also relates to the following.

[15]

[0043] A method for manufacturing the crystal according to any one of [1] to [5], which comprises a step of adding a seed crystal to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidine-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid, and a good solvent.

[16]

[0045] The manufacturing method according to

[15] , which comprises the following steps: adding a poor solvent to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidine-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid, and a good solvent, adding a seed crystal, and then further adding a poor solvent.

[0046] Effects of the invention

[0047] A crystal containing equimolar amounts of a pyrrolidine compound A and phosphoric acid is the following crystal: the solvent used in obtaining the crystal does not remain; it has excellent thermal stability; it has little weight change and is stable with respect to humidity; it does not deliquesce; it has excellent chemical stability; it does not contain a compound that may have an adverse effect on a living body from the viewpoint of safety, and since the crystal can be obtained with good reproducibility by an industrially suitable method, it is useful as a pharmaceutical raw material. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 Figure 1 It is a figure showing the result of powder X-ray diffraction measurement of the crystal of the present invention.

[0049] Figure 2 Figure 2 It is a figure showing the result of differential scanning calorimetry measurement of the crystal of the present invention.

[0050] Figure 3 Figure 3 It is a figure showing an ORTEP diagram of single crystal X-ray diffraction measurement based on the molecules in the crystal of the present invention.

[0051] Figure 4 Figure 4 It is a figure showing a packing diagram (a-axis projection diagram) of single crystal X-ray diffraction measurement based on the molecules in the crystal of the present invention.

[0052] Figure 5-1 Figure 5-1 It is a figure showing the particle size distribution of the crystal obtained under Condition 1 in Experimental Example 7.

[0053] Figure 5-2 Figure 5-2 It is a figure showing the particle size distribution of the crystal obtained under Condition 3 in Experimental Example 7. DETAILED DESCRIPTION OF THE INVENTION

[0054] The present invention relates to a crystal containing equimolar amounts of a pyrrolidine compound A represented by the following formula and phosphoric acid, a pharmaceutical composition containing the crystal as an active ingredient, and the like.

[0055] [Chemical Formula 1]

[0056]

[0057] The pyrrolidine compound A and / or phosphoric acid in the crystal of the present invention include compounds labeled with isotopes (for example, 3 H, 13 C, 14 C, 15 N, 18 F, 32 P, etc.) and deuterium-converted forms. ​​​​​​​​​​​​

[0058] In the present invention, in the case of a crystal containing equimolar amounts of pyrrolidine compound A and phosphoric acid, no other molecules such as the solvent used for obtaining the crystal remain, and pyrrolidine compound A and phosphoric acid form a crystal in a molar ratio of 1:1, that is, 1 molar equivalent of phosphoric acid is contained relative to pyrrolidine compound A.

[0059] As a preferred mode of the crystal of the present invention, there can be mentioned a co-crystal in which pyrrolidine compound A and phosphoric acid are not formed into a salt, but pyrrolidine compound A and phosphoric acid are bonded by non-ionic bonds and / or non-covalent bonds in a molar ratio of 1:1. In one mode, the crystal of the present invention exhibits Figure 1 the powder X-ray diffraction pattern shown, and in terms of the diffraction angle represented by 2θ, characteristic peaks can be mentioned at 5.7°, 11.5°, 13.9°, 19.0° and / or 21.9° (±0.2° respectively). More specifically, in terms of the diffraction angle represented by 2θ, peaks are present at 5.7°, 11.5°, 13.9°, 17.4°, 19.0°, 20.4° and / or 21.9° (±0.2° respectively). More specifically, in terms of the diffraction angle represented by 2θ, peaks are present at 5.7°, 7.4°, 11.5°, 12.3°, 13.9°, 17.4°, 19.0°, 20.4° and / or 21.9° (±0.2° respectively). Particularly specifically, in terms of the diffraction angle represented by 2θ, the peaks described in Table 2 below are present (hereinafter, the crystal having these peaks may sometimes be referred to as a B-type crystal or a phosphoric acid B-type crystal). In another mode, the above B-type crystal shows Figure 2 the differential scanning calorimetry (hereinafter, sometimes referred to as DSC) curve shown, and has an endothermic peak at 230°C to 240°C.

[0060] The crystal of the present invention is a crystal in which the residual solvent is below the reference value specified by the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (hereinafter referred to as ICH), and has an advantageous effect in this regard. In addition, as other advantages, it can be mentioned that it is a crystal in which organic impurities, inorganic impurities, residual metals, residual solvents, genotoxic impurities, etc. are below the reference values specified by the guiding principles of ICH.

[0061] In addition, as the crystal of the present invention, it is preferably manufactured into a crystal having a mode particle size of 7 μm or more for non-aggregated crystals, and more preferably 7 to 15 μm, particularly preferably 8 to 12 μm. The mode particle size refers to the particle size at which the volume% of the particle size distribution takes a maximum value. In the manufacturing method of the crystal of the present invention, by making at least 70% or more, preferably 80% or more, and more preferably 90% or more of all the crystals have such a mode particle size, the operability such as the ease of crystal filtration is improved.

[0062] The crystal of the present invention can be obtained, for example, by reacting 1 to 10 moles, preferably 1 to 5 moles of phosphoric acid with respect to 1 mole of the pyrrolidine compound A that can be manufactured according to the method described in Example 19 of Patent Document 1. In addition, it can also be manufactured by the method shown in the examples described later.

[0063] The solvent used to obtain the crystal of the present invention can be appropriately selected. For example, a good solvent or a poor solvent can be used alone or in an appropriate combination. As the good solvent, any solvent with a high solubility of the pyrrolidine compound A can be used without limitation. For example, ketones (e.g., acetone, 2-butanone, etc.), esters (e.g., ethyl acetate, methyl acetate, etc.), alcohols (e.g., methanol, ethanol, isopropyl alcohol, etc.), and mixtures of these solvents can be cited. As the poor solvent, any solvent with a low solubility of the pyrrolidine compound A can be used without limitation. For example, water, alkanes (e.g., hexane, heptane, etc.), aromatic hydrocarbons (e.g., benzene, toluene, etc.), ethers (e.g., diethyl ether, dimethyl ether, diisopropyl ether, etc.), and mixtures of these solvents can be cited.

[0064] As one mode of the method for obtaining the crystal of the present invention, the following method can be cited: adding phosphoric acid to a mixture of the pyrrolidine compound A and a good solvent, adding a seed crystal to the obtained mixture, and performing filtration. Preferably, the following method can be cited: dissolving the pyrrolidine compound A in a good solvent, adding phosphoric acid, adding a seed crystal to the obtained mixture, and then performing filtration. More preferably, the following method can be cited: dissolving the pyrrolidine compound A in ethyl acetate or ethanol, adding phosphoric acid, adding a seed crystal to the obtained mixture, and then performing filtration.

[0065] Alternatively, as another method, the following method can be cited: Phosphoric acid is added to a mixture of a pyrrolidine compound A and a good solvent, a poor solvent is added to the resulting mixture, and filtration is carried out. Preferably, the following method can be cited: The pyrrolidine compound A is dissolved in a good solvent, phosphoric acid is added, a poor solvent is added to the resulting mixture, and then a seed crystal is added, followed by filtration. More preferably, the following method can be cited: The pyrrolidine compound A is dissolved in a good solvent, phosphoric acid is added, a poor solvent is added to the resulting mixture, a seed crystal is added, and then a poor solvent is further added, followed by filtration. Even more preferably, the following method can be cited: The pyrrolidine compound A is dissolved in a good solvent, phosphoric acid is added, water is added to the resulting mixture, a seed crystal is added, and then water is further added, followed by filtration.

[0066] As suitable combinations and ratios when using a good solvent and a poor solvent in combination, for example, ethanol:toluene = 1:9, ethanol:diisopropyl ether = 3:7, acetone:toluene = 3:7, etc. can be cited. As examples of suitable good solvents when using a good solvent and water in combination, ethyl acetate, ethanol, etc. can be cited. When using a good solvent and a poor solvent (specifically water) in combination and adding water twice before and after adding the seed crystal, it is preferable that the amount of water added before adding the seed crystal (the first addition of water) is 3 times in volume ratio relative to the weight of the pyrrolidine compound A, and the amount of water added after adding the seed crystal (the second addition of water) is 4.5 times in volume ratio relative to the weight of the pyrrolidine compound A. As another method, the following method can be cited: The amount of water added is 5 to 10 times in volume ratio relative to the weight of the pyrrolidine compound A, and it is added in batches before and after adding the seed crystal. More preferably, the amount of water is 6 to 9 times in volume ratio relative to the weight of the pyrrolidine compound A, and even more preferably 7.5 times. Alternatively, as another example, the volume ratio of the amount of water added for the first time to the amount of water added for the second time is 1:1 to 1:2, preferably 2:3. As another example, water is added before adding the seed crystal to such an extent that 70% or more, preferably 80% or more of the crystals precipitate after adding the seed crystal and before adding water. In addition, the following example can be cited: The temperature when adding the seed crystal is set to 28 to 32 °C, preferably set to 30 °C. This temperature is also preferably selected as the temperature at which 70% or more, preferably 80% or more of the crystals precipitate after adding the seed crystal and before adding water.

[0067] The seed crystal of the crystal of the present invention can be obtained by the methods described in Example 3, Experimental Example 2, or Experimental Example 3 below. Alternatively, the crystals obtained by these methods can also be used as the seed crystal of the crystal of the present invention, and the seed crystal can be obtained by, for example, the methods described in Example 1 or 2 or the methods based on the methods described in Example 1 or 2.

[0068] The crystal of the present invention has human MC1R agonist activity, and thus can be used as an active ingredient of a melanocortin receptor 1 agonist. In addition, the crystal of the present invention and a pharmaceutical composition containing the same as an active ingredient are useful in treating or preventing various diseases whose conditions are expected to be improved by activation of MC1R. Examples of such diseases include one or more diseases selected from rheumatoid arthritis, gouty arthritis, osteoarthrosis, inflammatory bowel disease, systemic sclerosis, psoriasis, fibrosis, porphyria (e.g., erythropoietic protoporphyria, etc.), systemic lupus erythematosus, melanoma, skin cancer, vitiligo, alopecia, pain, ischemia / reperfusion injury, inflammatory diseases of the brain, hepatitis, sepsis / septic shock, nephritis, transplantation, deterioration of HIV disease, vasculitis, uveitis, retinitis pigmentosa, age-related macular degeneration, microbial infection, coeliac disease, nephrotic syndrome, and melanoma infiltration. In particular, it is useful in treating or preventing one or more diseases selected from systemic sclerosis, psoriasis, porphyria, melanoma, skin cancer, vitiligo, alopecia, retinitis pigmentosa, age-related macular degeneration, and nephrotic syndrome. Especially, it is useful in treating or preventing one or more diseases selected from systemic sclerosis, porphyria, melanoma, vitiligo, retinitis pigmentosa, age-related macular degeneration, and nephrotic syndrome.

[0069] The pharmaceutical composition containing the crystal of the present invention as an active ingredient can be obtained by mixing the crystal of the present invention with pharmaceutically acceptable additives such as excipients, disintegrants, binders, lubricants, coating agents, pigments, diluents, bases, and isotonic agents.

[0070] For the crystal of the present invention and the pharmaceutical composition containing the same as an active ingredient, after being prepared into an appropriate administration form (e.g., powder, injection, tablet, capsule, and topical agent, etc.), they can be administered to a patient by an appropriate administration method corresponding to the administration form (e.g., intravenous administration, oral administration, and transdermal administration, etc.). The term "patient" in the present invention is an individual who is the subject to be prevented or treated by the crystal of the present invention, preferably a mammal, more preferably a human.

[0071] Regarding the dosage, it can be determined based on considerations of these factors or other factors such as the patient's age, weight, general health status, gender, diet, administration time, administration method, excretion rate, combination of drugs, and the degree of the patient's medical condition being treated at the time of administration. The crystals of the present invention and the pharmaceutical composition containing the same as an active ingredient have low toxicity and can be safely used. The daily dosage (i.e., the effective amount) varies depending on the patient's condition, weight, administration route, etc. For example, in the case of parenteral administration, it is preferably administered in an amount of about 0.0001 to 1000 mg / person / day, more preferably about 0.001 to 1000 mg / person / day, and particularly preferably 0.01 to 500 mg / person / day. Additionally, in the case of oral administration, it is preferably administered in an amount of about 0.0001 to 1000 mg / person / day, more preferably 0.01 to 500 mg / person / day.

[0072] In the present invention, "prevention" refers to the act of administering the crystals of the present invention or a pharmaceutical composition containing the same to an individual who has not developed a disease, illness, or symptom. Additionally, "treatment" refers to the act of administering the crystals of the present invention or a pharmaceutical composition containing the same to an individual who has developed a disease, illness, or symptom. Therefore, the act of administering to an individual who has developed a disease, illness, or symptom in order to prevent the deterioration of symptoms, prevent the onset of a disease, or prevent the recurrence of a disease is a form of "treatment".

[0073] Examples

[0074] Hereinafter, the present invention will be described in detail by way of examples and experimental examples, but the present invention is not limited by any of them. It should be noted that "equivalent" refers to "molar equivalent".

[0075] Example 1 Synthesis of the crystals of the present invention

[0076] [Chemical formula 2]

[0077]

[0078] Dissolve compound 1 (26.18 g) in dichloromethane (207 mL), add compound 2 (4.4 mL) and acetic acid (3.56 mL) thereto, and stir at room temperature for 30 minutes. Then, further add sodium triacetoxyborohydride (13.2 g) and stir at room temperature for 1 hour. After adding a saturated aqueous sodium bicarbonate solution and stirring, extract with dichloromethane. Wash the obtained organic layer with a saturated brine solution, dry with magnesium sulfate, and concentrate under reduced pressure. Purify the residue by NH silica gel column chromatography (hexane:ethyl acetate = 75:25 to 55:45), and then by silica gel column chromatography (chloroform:methanol = 100:0 to 95:5) to obtain the compound as a colorless powder3 (24.25 g) (MS(ESI): m / z 690 [M+H] + )。

[0079] Dissolve compound 3 (24.24 g) in methanol (240 mL), add aqueous sodium hydroxide solution (2 mol / L, 70.2 mL) thereto, and stir at room temperature for 19 hours. Then, further add aqueous hydrochloric acid solution (2 mol / L, 74 mL), and then concentrate the reaction solution under reduced pressure. Add water and ethyl acetate to the concentrated residue, stir, and extract with ethyl acetate. Wash the obtained organic layer with phosphate buffer solution (0.1 mol / L, 300 mL) and brine, dry over magnesium sulfate, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography (chloroform:methanol = 100:0 to 90:10), dissolve it in ethyl acetate, add phosphate buffer solution (0.1 mol / L, 200 mL), stir at room temperature, and extract with ethyl acetate. Wash the organic layer with water and saturated brine, dry over magnesium sulfate, and concentrate under reduced pressure to obtain compound 4 (23.7 g) (MS(ESI): m / z 676 [M+H] + )。

[0080] Dissolve compound 4 (135 mg) in ethanol (0.7 mL), and add a small amount of the crystal of the present invention as a seed crystal. Add a phosphoric acid solution obtained by dissolving phosphoric acid (25 mg) in ethanol (0.5 mL) thereto, further add ethanol (0.2 mL), and stir at room temperature overnight. Filter out the precipitated crystals, wash with ethyl acetate (0.6 mL), and dry under reduced pressure at 50 °C for 4 hours to obtain 112.7 mg of the crystal of the present invention. Confirm the presence or absence of residual solvent by 1H-NMR, and no residual solvent was observed. The measurement results of elemental analysis are shown in Table 1 below.

[0081] [Table 1]

[0082] Table 1 <Elemental analysis measurement results>

[0083] C H N P Measured value 55.21 6.14 5.31 3.96 Theoretical value 55.49 6.29 5.39 3.97

[0084] (The theoretical values represent the theoretical values when 0.3 equivalents of water are attached.)

[0085] [Powder X-ray diffraction (hereinafter sometimes referred to as XRPD) measurement]

[0086] Use a powder X-ray diffraction measurement device X’PertPro (manufactured by PANalytical B.V.) to perform measurement under the following conditions.

[0087] X-ray generating device: X-ray tube (anticathode: copper, tube voltage: 45 kV, tube current: 40 mA)

[0088] Incident optical system: focusing condenser

[0089] Light-receiving optical system: high-speed semiconductor array detector (X-Celerator), expanded light-receiving side arm

[0090] Sample stage: HTS sample stage (vibrating with a width of 4 mm in the X-axis direction)

[0091] Number of accumulations: 5 times (the incident angles are changed to -2, -1, 0, 1, and 2° respectively)

[0092] Measurement range: 2θ = 3 to 40°

[0093] Scanning speed: 0.668451° / second

[0094] Interval: 0.0167°

[0095] The results are shown in Figure 1 . When the peak intensity at a diffraction angle of 5.7° indicated by 2θ is set to 100, the peaks with a relative peak intensity of 5 or more are shown in Table 2 below.

[0096] [Table 2]

[0097] Table 2

[0098]

[0099] <Differential scanning calorimetry (DSC) measurement>

[0100] Using a differential scanning calorimetry measurement device X-DSC7000 (SII NanoTechnology Inc.), the measurement was carried out under the following conditions.

[0101] Heating rate: 10 °C / min (25 °C to 300 °C)

[0102] Atmosphere: nitrogen 100 mL / min

[0103] The results are shown in Figure 2 . An endothermic peak was observed at approximately 230 °C to 240 °C.

[0104] <Single crystal X-ray diffraction measurement>

[0105] Add about half of the crystals of the present invention with a spatula (small) to 2 mL of ethanol, dissolve, and let stand at room temperature for 4 days for crystallization. Using a single crystal X-ray diffractometer R-AXIS RAPID / R (Rigaku Corporation) (CuKα line), the lattice constants of the obtained crystals were determined and the diffraction peak intensities were measured at 23 °C. Then, the phase was determined using the direct method, and the structure was refined using the full matrix least squares method to analyze the structure. The obtained crystallographic data and the results of the crystal structure analysis are shown in Table 3. It should be noted that the reliability factor (R value) is 3.06%, and various other parameters also show that the crystal structure analysis is a highly reliable analysis result.

[0106] [Table 3]

[0107] Table 3 Crystallographic data and results of crystal structure analysis

[0108]

[0109] The ORTEP diagram of the molecules in the crystals of the present invention is shown in Figure 3 , and the packing diagram is shown in Figure 4 . In the crystal, one molecule of pyrrolidine compound A and one molecule of phosphoric acid each independently exist in the asymmetric unit.

[0110] The absolute configuration of the crystals of the present invention was verified, and as a result, the Flack parameter was 0.02(3). Therefore, it was confirmed that the crystals of the present invention are composed of one molecule of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidine-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid shown in Figure 3 and one molecule of phosphoric acid as the structural units.

[0111] The guidelines issued by the US Food and Drug Administration (FDA) (Regulatory Classification of Pharmaceutical Co-Crystals Guidance for Industry) state that a co-crystal is a crystal that contains two or more different molecules in the crystal lattice and these molecules are bonded by non-ionic and / or non-covalent bonds. It has been confirmed by the above crystal structure analysis that there is no covalent bond between pyrrolidine compound A and phosphoric acid. Therefore, a study was conducted on whether ionic interactions were found between pyrrolidine compound A and phosphoric acid.

[0112] Since phosphoric acid is a Brønsted acid, in the case where an ionic interaction with pyrrolidine compound A is considered, pyrrolidine compound A must accept a proton as a Brønsted base. In the case of pyrrolidine compound A, the three nitrogens (N18, N19, N20) are the ones that can potentially accept a proton from phosphoric acid. Here, all the hydrogen bond sites in the crystal of the present invention are shown in Table 4.

[0113] [Table 4]

[0114] Table 4 Hydrogen bond sites in the crystal of the present invention

[0115]

[0116] (1) N18

[0117] N18 is an sp3 hybridized nitrogen, which is the nitrogen with the highest basicity in pyrrolidine compound A and can potentially accept a proton from phosphoric acid. However, it is judged that N18 (more precisely, the hydrogen atom bonded to N18) forms a hydrogen bond with the oxygen (O16) of the carboxylic acid of the adjacent pyrrolidine compound A (symmetry operators: -X + 1 / 2 + 2, -Y + 2, Z + 1 / 2), and thus the proton is donated. Therefore, N18 cannot accept a proton from phosphoric acid. At the same time, the interatomic distance between N18 and any oxygen of the adjacent phosphoric acid exceeds the sum of the van der Waals radii, so it is judged that there is no phosphoric acid within the distance where an ionic interaction can occur.

[0118] (2) N19

[0119] N19 is an sp2 hybridized nitrogen, so its basicity is weak and it is considered that a salt cannot be formed. At the same time, the interatomic distance between N19 and the oxygen (O13) of the nearest phosphoric acid is the sum of the van der Waals radii, so it is judged that it is not within the distance where an ionic interaction can occur.

[0120] (3) N20

[0121] N20 is also an sp2 hybridized nitrogen, so its basicity is weak and it is considered that a salt cannot be formed. At the same time, the interatomic distance between N20 and any oxygen of the adjacent phosphoric acid also exceeds the sum of the van der Waals radii, so it is judged that there is no phosphoric acid within the distance where an ionic interaction can occur.

[0122] Based on the above results, it can be seen that the binding between the nitrogen (N18, N19, N20) of pyrrolidine compound A and phosphoric acid in the crystal of the present invention is not based on ionic interaction, pyrrolidine compound A and phosphoric acid do not form a salt, and it is judged that the crystal of the present invention is a co-crystal. In this specification, the crystal obtained in this example is referred to as Type B crystal.

[0123] Example 2 Synthesis of the crystal of the present invention (2)

[0124] [Chemical formula 3]

[0125]

[0126] To a solution of the compound 3 (276 mg) in ethanol (1.4 mL) was added 1,2-ethanedisulfonic acid hydrate (38 mg), and the mixture was stirred at room temperature for 40 minutes. The crystals were filtered out and washed twice with ethanol (0.84 mL). Drying was carried out at 40 °C or below to obtain the compound 5 (178 mg). To a suspension of the compound 5 (37.1 kg) in ethyl acetate (167.5 kg) were successively added at room temperature a solution of potassium carbonate (6.5 kg) in water (148.3 L) and water (36.8 L). The mixture was stirred at room temperature for 15 minutes. The aqueous layer was removed, and the organic layer was washed twice with water (186 L). Ethyl acetate (67.2 kg) was added, and the insoluble matter was filtered. After concentration to 78 L, ethanol (146.5 kg) was added, and the mixture was concentrated to 78 L. Ethanol (146.9 kg) was added, and the mixture was concentrated to 56 L. Dilution was carried out with ethanol (44 kg), and a 24% aqueous sodium hydroxide solution (8.7 kg) and water (30.1 kg) were successively added at room temperature. The mixture was stirred at 40 °C for 5 hours to obtain the compound 4 . At 30 °C, to a solution of the compound 4 were successively added a solution of phosphoric acid (12.0 kg) in water (55.7 L) and water (55.7 L). Seeds of the crystals of the present invention (928 g) were added, and the mixture was stirred for 14 hours. Water (167.0 L) was added, and after stirring for 4 hours, the mixture was cooled to 25 °C. The solid was filtered out and washed with water (182 L). The solid was dried at 50 °C or below to obtain the crystals of the present invention (35.2 kg). These crystals have excellent operability such as filterability.

[0127] Experimental Example 1 Crystallization Study of Pyrrolidine Compound A

[0128] Using pyrrolidine compound A, the following studies were conducted: crystallization studies through long-term storage using 25 single solvents and 44 mixed solvents; crystallization studies through 1-month stirring using 96 mixed solvents (prepared by using 2 anti-solvents and 12 good-solvents and changing the mixing ratio of the solvents); and crystallization studies through grinding using 26 added solvents. As a result, crystals could be obtained through the crystallization studies based on long-term storage and 1-month stirring. The obtained crystals were subjected to powder X-ray diffraction measurement. As a result, in all the crystals, the peaks were consistent and they were of the same crystal form. The crystallization conditions were studied, and in all the conditions where crystals could be obtained, solvents remained in the crystals. The process of drying the crystals was also studied, but it was not possible to reduce the residual solvents below the residual solvent reference values established by ICH.

[0129] Therefore, although pyrrolidine compound A as a free body crystallized, the solvents used when obtaining the crystals remained, and in addition, the chemical stability was low, so it could not be used as a pharmaceutical raw material.

[0130] Experimental Example 2 Crystallization study of a mixture containing pyrrolidine compound A (1)

[0131] Approximately 900 mg of pyrrolidine compound A was dissolved in 30 mL of tetrahydrofuran, and 100 μL (about 3 mg / vial) was dispensed into each vial of a 96-well plate. In addition, 21 acids including phosphoric acid, hydrochloric acid, L(-)-malic acid, L(+)-tartaric acid, maleic acid, sulfuric acid, and malonic acid, and 8 bases including sodium hydroxide and L-arginine (hereinafter sometimes referred to as counter compounds) were dissolved in 8 solvents described below, and 45 μL of the resulting 0.1 mol / L solution (for some acids, 90 μL of a 0.05 mol / L solution) was dispensed into each vial. It was left open for one day and night to allow the solvent to evaporate, and then dried under reduced pressure for 4 hours. After 250 μL of 8 solvents including ethyl acetate, acetone, and toluene were dispensed into each vial, it was stirred ultrasonically for 5 minutes, sealed, and stirred at room temperature for 6 days. For the vials with precipitates, the precipitates were filtered out and XRPD was measured. For the vials without precipitates, the solvent was allowed to evaporate at room temperature, and if a solid was observed, it was filtered out and XRPD was measured. The XRPD measurement apparatus and measurement conditions were the same as in Example 1.

[0132] From vials containing combinations of phosphoric acid and ethyl acetate, L(-)-malic acid and toluene, L(+)-tartaric acid and acetone, and maleic acid and toluene, phosphoric acid A-type crystals, L(-)-malic acid D-type crystals, L(+)-tartaric acid E-type crystals, and maleic acid F-type crystals were obtained respectively. It should be noted that in addition to these, there were also vials in which only the crystals of the paired compounds, that is, the crystals containing no pyrrolidine compound A, could be confirmed. On the other hand, using all 8 solvents, no crystals could be obtained from the vials containing hydrochloric acid. In addition, using all solvents, no crystals could be obtained from the vials containing sulfuric acid, malonic acid, and L-arginine either.

[0133] Experimental Example 3 Crystallization Study of a Mixture Containing Pyrrolidine Compound A (2)

[0134] Dissolve approximately 800 mg of pyrrolidine compound A in 40 mL of tetrahydrofuran, and dispense 100 μL (approximately 2 mg / vial) into each vial of a 96-well plate. In addition, dissolve 22 acids including phosphoric acid, hydrochloric acid, L(-)-malic acid, L(+)-tartaric acid, maleic acid, sulfuric acid, and malonic acid, and 8 bases including sodium hydroxide and L-arginine (hereinafter sometimes referred to as paired compounds) in 12 solvents described below, and dispense 30 μL of the resulting 0.1 mol / L solution (for some acids, 60 μL of 0.05 mol / L solution) into each vial. After dispensing, blow nitrogen to evaporate the solvent. Dispense 200 μL of 12 solvents including a mixed solvent of ethanol and toluene with a mixing ratio of 3:7 and 1:9, a mixed solvent of ethanol and diisopropyl ether with a mixing ratio of 3:7, a mixed solvent of acetone and toluene with a mixing ratio of 3:7 and 1:9, and a mixed solvent of ethyl acetate and heptane with a mixing ratio of 3:7 into each vial, seal, and stir at room temperature for 3 days. For the vials with precipitates, filter out the precipitates and measure XRPD. For the vials without precipitates, evaporate the solvent at room temperature, and if a solid is observed after about 1 month, collect the solid and measure XRPD. The XRPD measurement apparatus and measurement conditions are the same as those in Example 1.

[0135] The vials containing the crystal combinations obtained are shown in Table 5 below. In the table, "-" indicates that no crystal was obtained, and A, B, etc. indicate the obtained A-type crystal, B-type crystal, etc., respectively. That is, it is shown that the XRPD patterns of the F-type maleate crystals obtained this time are consistent with those of the F-type maleate crystals obtained in Experimental Example 2, and the B-type phosphate crystals obtained this time are different crystals from the A-type phosphate crystals obtained in Experimental Example 2. It should be noted that, similar to Experimental Example 2, in addition to the above, there are also vials in which only the crystals of the paired compounds could be confirmed. On the other hand, using all 12 solvents, no crystals could be obtained from the vials containing hydrochloric acid. In addition, regarding L(-)-malic acid and L(+)-tartaric acid, crystals were obtained in Experimental Example 2, but no crystals could be obtained this time using all 12 solvents.

[0136] [Table 5]

[0137] Table 5 Results of the crystallization study (2) of the mixture containing pyrrolidine compound A

[0138]

[0139] Experimental Example 4 Crystallization study (3) of the mixture containing pyrrolidine compound A

[0140] Among the crystals obtained in Experimental Examples 2 and 3, for the crystals other than the A-type phosphate crystals and the B-type phosphate crystals, the reproducibility was confirmed. As a result, the D-type L(-)-malic acid crystals, the E-type L(+)-tartaric acid crystals, the J-type sulfuric acid crystals, and the K-type L-arginine crystals could not be obtained again. On the other hand, crystals could be obtained again for the combination of maleic acid and malonic acid.

[0141] <L(-)-Malic acid, L(+)-tartaric acid, and sulfuric acid>

[0142] Dissolve approximately 80 mg of pyrrolidine compound A in 4 mL of tetrahydrofuran, and dispense 100 μL (approximately 2 mg / vial) into each vial of a 96-well plate. In addition, dispense 30 μL each of 0.1 mol / L solutions obtained by dissolving L(-)-malic acid, L(+)-tartaric acid, and sulfuric acid in the solvents described below into each vial. After dispensing, evaporate the solvent by blowing nitrogen, and then dispense 200 μL of toluene into the vial containing L(-)-malic acid, 200 μL of acetone into the vial containing L(+)-tartaric acid, and a mixed solvent with a mixing ratio of 3:7 of ethyl acetate and heptane into the vial containing sulfuric acid, and seal. For each sample, 8 were prepared. A total of 24 vials were stirred at room temperature for 7 days, but no precipitates were obtained in all the vials.

[0143] <L-arginine>

[0144] Weigh approximately 65 mg of pyrrolidine compound A and dissolve it in 0.3 mL of ethanol and 2.1 mL of toluene at room temperature. Weigh approximately 19 mg of L-arginine, dissolve it in 0.6 mL of ethanol and 0.6 mL of water, and add it to the solution of the above pyrrolidine compound A. Since it is a solution, the solvent is evaporated by blowing nitrogen gas, and as a result, a sugar-like substance containing some white powder is obtained. It is dissolved again in 0.3 mL of ethanol, and 0.6 mL of diisopropyl ether is added dropwise. As a seed crystal, the L-arginine K-type crystal obtained in Experimental Example 3 is added, but it dissolves, so 1.2 mL of toluene is added and stirred overnight. Since it is a solution, the solvent is evaporated by blowing nitrogen gas, and the residue is subjected to XRPD measurement, and the result is amorphous. Add 0.6 mL of heptane thereto, stir at room temperature overnight, and then perform microscopic observation. No crystal components are observed.

[0145] <Maleic acid F-type crystal>

[0146] Weigh approximately 65 mg of pyrrolidine compound A and dissolve it in 0.5 mL of toluene at room temperature. Add thereto a substance obtained by dissolving approximately 13 mg of maleic acid in 75 μL of ethanol and a small amount of the previously obtained maleic acid F-type crystal as a seed crystal. As a result, the reaction solution becomes gel-like (agar-like) and cannot be stirred. Further add 1.0 mL of toluene and vigorously rub it with a spatula. As a result, it becomes a suspension. The filtered crystal is dried under reduced pressure at 40 °C for 5 hours to obtain 57 mg of maleic acid F-type crystal. It was confirmed by 1H-NMR that 0.1 equivalent of toluene remained in the obtained crystal.

[0147] <Malonic acid G-type crystal>

[0148] Weigh approximately 325 mg of pyrrolidine compound A and dissolve it in 1 mL of acetone at room temperature. Weigh 55 mg of malonic acid and dissolve it in 0.5 mL of acetone at room temperature, and add it to the solution of the above pyrrolidine compound A. Add 6 mL of toluene dropwise to this solution. Add a small amount of the malonic acid I-type crystal described below as a seed crystal and stir at room temperature for two nights. Filter the whole amount and dry it under reduced pressure at 40 °C for 3.5 hours to obtain 317 mg of crystal. It was confirmed by 1H-NMR that the obtained crystal contained 1 equivalent of toluene. In order to replace toluene with water, it was stored for 72 hours under the conditions of 25 °C and 70% RH using a Dynamic Vapour Sorption (DVS) apparatus, and as a result, amorphousization was observed.

[0149] <Malonic acid H-type crystal>

[0150] Weigh approximately 65 mg of pyrrolidine compound A and dissolve it in 2 mL of a mixed solvent of ethyl acetate and heptane with a mixing ratio of 3:7 at room temperature. Weigh 11 mg of malonic acid and dissolve it in 0.3 mL of ethyl acetate at room temperature, and add it to the solution of the above pyrrolidine compound A. Since a hard rubbery substance was formed, 3 mL of ethyl acetate was added to make a suspension. After stirring at room temperature for 3 days, the solvent was evaporated by blowing nitrogen gas. 3 mL of ethyl acetate was added to the dried solid and stirred, and 1.2 mL of heptane was slowly added. When a small amount of the malonic acid G-type crystals obtained in Experimental Example 3 was added as a seed crystal, precipitates were rapidly generated. After stirring at room temperature for two nights, the whole amount was filtered and dried under reduced pressure at 40 °C for 3.5 hours to obtain 38 mg of crystals.

[0151] <Malonic acid type I crystals>

[0152] Weigh approximately 65 mg of pyrrolidine compound A and dissolve it in 2.3 mL of a mixed solvent of acetone and toluene with a mixing ratio of 1:9 at room temperature. Weigh 11 mg of malonic acid and dissolve it in 0.1 mL of acetone at room temperature, and add it to the solution of the above pyrrolidine compound A. Stir at room temperature for 3 nights, filter the whole amount, and dry it under reduced pressure at 40 °C for 2 hours to obtain 51 mg of crystals. It was confirmed by 1H-NMR that the obtained crystals contained 1 equivalent of toluene. Since these crystals might be a toluene adduct, for the purpose of confirming whether solvent-free crystals could be obtained by further drying under reduced pressure, approximately 5 mg was dried under reduced pressure at 60 °C for 4 hours, and as a result, approximately 0.5 equivalent of toluene remained, and it was confirmed that the crystal form did not change.

[0153] Example 3 Synthesis of the crystals of the present invention (3)

[0154] Add the amounts of pyrrolidine compound A and phosphoric acid shown in Table 6 below to the solvents shown below and stir for the times shown below. The results are shown in Table 6 below. The XRPD measurement apparatus and measurement conditions are the same as those in Example 1. The XRPD results show that A is the same crystal as the phosphoric acid A-type crystals obtained in Experimental Example 2, and B is the same crystal as the phosphoric acid B-type crystals obtained in Experimental Example 3. In addition, the phosphoric acid B-type crystals are the same crystals as the crystals of the present invention obtained in Example 1.

[0155] [Table 6]

[0156] Table 6

[0157]

[0158] In method (a), a mixture of A-type crystals and B-type crystals was obtained. From the results of elemental analysis, it was determined that the crystals contained 1.5 equivalents of phosphoric acid. In method (f) using an acetone / toluene solvent, new C-type crystals were obtained. From the results of elemental analysis, it was determined that the C-type crystals contained 2 equivalents of phosphoric acid. In addition, regardless of which method was used, A-type crystals could not be obtained alone.

[0159] In methods (a), (b), (c), (e) and (f), an amorphous substance or a sugary substance was immediately formed after adding phosphoric acid, but by stirring the amorphous substance at room temperature for 7 to 10 days, or by frequently rubbing the sugary substance with a spatula for a long time, crystals could be obtained. In addition, in method (d), a suspension of crystals was obtained 1 day after the start of crystallization, but it became a sugary substance after continuous stirring at room temperature for 4 days. Furthermore, the solvent was evaporated with nitrogen, and it was stirred at room temperature in ethanol overnight, and as a result, B-type crystals were obtained.

[0160] Experimental Example 5 Synthesis of hydrochloride of pyrrolidine compound A

[0161] [Chemical formula 4]

[0162]

[0163] To a chloroform (157 mL) solution of compound 1 (19.43 g) was added compound 2 (3.3 mL) and acetic acid (2.7 mL). After stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (10.0 g) was added, and the mixture was stirred at room temperature for 20 hours. After adding a saturated aqueous sodium bicarbonate solution and stirring, extraction was carried out with chloroform. The obtained organic layer was washed with a saturated aqueous sodium bicarbonate solution and saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (hexane:ethyl acetate = 75:25 to 55:45), then by silica gel column chromatography (chloroform:methanol = 100:0 to 95:5) and silica gel column chromatography (chloroform:methanol = 99:1 to 96:4) to obtain compound 3 (21.92 g) (MS(ESI): m / z 690 [M+H] + ).

[0164] To compound 3To a solution of methanol (200 mL, 21.91 g) was added an aqueous sodium hydroxide solution (2 mol / L, 63.6 mL), and the mixture was stirred at room temperature for 3 hours. After adding an aqueous hydrochloric acid solution (2 mol / L, 63.6 mL), the reaction solution was concentrated under reduced pressure. Water and ethyl acetate were added to the concentrated residue, and after stirring, extraction was carried out with ethyl acetate. The obtained organic layer was washed with a saturated brine solution, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform:methanol = 98:2 to 90:10), then dissolved in ethyl acetate, and a phosphate buffer solution (0.1 mol / L, 300 mL) was added, and the mixture was stirred at room temperature for 3 hours. After extraction with ethyl acetate, the organic layer was washed with a saturated brine solution, dried over magnesium sulfate, and concentrated under reduced pressure, whereby the compound was obtained as a colorless powder 4 (17.40 g) (MS(ESI): m / z 676 [M+H] + )

[0165] To a solution of the compound 4 (17.40 g) in ethyl acetate (250 mL) was added a hydrochloric acid-ethyl acetate solution (4 mol / L, 31.8 mL), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, ether was added, and after stirring, filtration was carried out and drying was carried out under reduced pressure, whereby the hydrochloride (17.43 g) of the compound 4 was obtained as a colorless powder (MS(ESI): m / z 676 [M+H] + )

[0166] Experimental Example 6 Comparison of the amorphous form of the hydrochloride of pyrrolidine compound A, the amorphous form of pyrrolidine compound A, the crystal of pyrrolidine compound A, and the crystal of the mixture containing pyrrolidine compound A for which reproducibility has been confirmed

[0167] Since the hydrochloride of pyrrolidine compound A described in Example 19 of Patent Document 1 is amorphous and the crystal of the hydrochloride of pyrrolidine compound A has not been obtained in subsequent studies (see Experimental Examples 2 and 3 above), the amorphous hydrochloride of pyrrolidine compound A was used for the comparative test. In addition, in the comparative test results shown below, the hydrochloride is the hydrochloride of pyrrolidine compound A, and the results of the compound obtained by the method described in Experimental Example 5 are shown.

[0168] In the comparative test results shown below, the free amorphous form is the free amorphous form of pyrrolidine compound A, and the results of the compound obtained by the method described in Example 1 are shown 4 of

[0169] In the comparative test results shown below, the free crystal is a crystal of the free pyrrolidine compound A. As shown in Experimental Example 1, for this crystal, it is impossible to reduce the residual solvent in the crystal below the reference value specified by the ICH, and it has been confirmed that this crystal is not a crystal suitable as a pharmaceutical active ingredient.

[0170] In the comparative test results shown below, for the phosphoric acid A + B type crystal, it represents the result of the crystal obtained by the method (a) described in Example 3; for the phosphoric acid B type crystal, it represents the result of the crystal of the present invention obtained by the method described in Example 1; for the phosphoric acid C type crystal, it represents the result of the crystal obtained by the method (f) described in Example 3; for the maleic acid F type crystal, it represents the result of the maleic acid F type crystal obtained by the method described in Experimental Example 4.

[0171] In the comparative test results shown below, for the malonic acid H type crystal, it represents the result of the malonic acid H type crystal obtained by the method described in Experimental Example 4. For malonic acid, in addition to the H type, G type and I type crystals can also be obtained, and these all contain toluene in the molecule. Toluene is a compound known to cause disorders to the central nervous system, etc. From the perspective of safety, it is not desirable to be included in a pharmaceutical active ingredient. It can be seen that the G type and I type crystals are not suitable as pharmaceutical active ingredient crystals, and therefore comparative tests were not conducted on them.

[0172] For each of the above crystals, the thermal stability, hygroscopicity / deliquescence, and chemical stability were evaluated.

[0173] <Thermal stability evaluation>

[0174] Using a thermogravimetry / differential thermal simultaneous measurement device TG / DTA7200 (SII NanoTechnology Inc.), the evaluation was carried out under the following conditions.

[0175] Heating rate: 10 K / min

[0176] Atmosphere: Nitrogen 200 mL / min

[0177] <Hygroscopicity / deliquescence evaluation>

[0178] Using a moisture adsorption measurement device DVS-1 or DVS-intrinsic (Surface Measurement Systems Limited), the evaluation was carried out in the following manner. The sample was taken into a dish whose tare weight had been previously calibrated, hung on the precision balance of the device, and the weight at the start of the measurement was precisely determined. The weight change when the humidity was changed stepwise over time was recorded, and the equilibrium weight at each humidity was determined. Taking the anhydride obtained by drying (0% RH) or the anhydrous substance converted from the initial moisture content confirmed by other methods as the reference, the change rate of the weight at each humidity was determined.

[0179] <Chemical stability evaluation>

[0180] The sample was stored for 1 week under sealed conditions at 60 °C and under the conditions of 60 °C and 75% RH. Using high performance liquid chromatography, based on the area percentage of each peak, the increase or decrease of related substances before and after storage was calculated, and the state after storage was further observed.

[0181] The results are shown in Table 7 below.

[0182] [Table 7]

[0183]

[0184] As reproducible solids in pyrrolidine compound A and mixtures containing the same, hydrochloride amorphous, free form amorphous, free form crystal, phosphoric acid type A + type B crystal, phosphoric acid type B crystal as a mode of the crystal of the present invention, phosphoric acid type C crystal, maleic acid type F crystal, malonic acid type G crystal, malonic acid type H crystal, and malonic acid type I crystal were found. Among these, for the free form crystal, the residual solvent could not be reduced to below the reference value specified by ICH. For the malonic acid type G crystal and malonic acid type I crystal, toluene was contained in the molecule, and from the viewpoint of safety, they were not suitable as crystals for pharmaceutical raw materials. The maleic acid type F crystal also could not completely remove toluene as the residual solvent. On the other hand, no residual solvent was confirmed in the crystal of the present invention, and toluene was not contained in the molecule. Therefore, from the viewpoint of safety, it was considered to be a crystal without problems.

[0185] In addition, hygroscopicity was observed in hydrochloride amorphous, phosphoric acid type C crystal, maleic acid type F crystal, and malonic acid type H crystal, but hygroscopicity was not observed in phosphoric acid type B crystal, the weight change at 90% RH was less than 1%, and even when stored for 1 week under the conditions of 60 °C and 75% RH, the increase in related substances was only 0.05%. It was a crystal that was stable to humidity and very stable chemically.

[0186] In addition, when the free amorphous substance and the crystalline forms of phosphoric acid type A + B type are heated to 130 °C, a weight change of more than 2% is observed in both cases. However, no weight change is observed for the crystalline form of phosphoric acid type B even at temperatures above 200 °C, indicating that it is a crystal with excellent thermal stability.

[0187] Experimental Example 7 Study on the precipitation conditions of the crystals of the present invention

[0188] [Chemical formula 5]

[0189]

[0190] Condition 1

[0191] To a solution of compound 5 (15.00 g) in ethyl acetate (67.75 g), an aqueous potassium carbonate solution (2.64 g of potassium carbonate, 75 mL of purified water) was added dropwise at 25 °C, and the mixture was stirred at 300 rpm for 30 minutes. The organic layer was separated, washed with purified water, and then concentrated under reduced pressure. Ethanol (59.03 g) was added, and the mixture was concentrated under reduced pressure until the volume became 27 mL. This operation was repeated twice.

[0192] To the resulting concentrated mixture, ethanol (24.32 g) was added until the volume became 45 mL. At 25 °C, a 24% aqueous sodium hydroxide solution (4.79 g) and purified water (12 mL) were added dropwise, and the mixture was stirred at 300 rpm for 6 hours. To the reaction mixture, an 85% aqueous phosphoric acid solution (6.61 g) and purified water (22.5 mL, 1.5 times the volume based on the weight of compound 4 were added dropwise over 5 minutes, and then purified water (90 mL, 6.0 times the volume based on the weight of compound 4 ) was added dropwise over 30 minutes. After the addition of purified water was completed, the mixture was stirred for 30 minutes, the reaction temperature was adjusted to 35 °C, and a seed crystal (0.3746 g, 0.025 times the weight based on the weight of compound 4 ) was added. Ten hours after the addition of the seed crystal, the reaction temperature was adjusted to 20 °C, and after further stirring for 30 minutes, the insoluble matter was filtered off and dried under reduced pressure to obtain the crystals of the present invention (7.48 g).

[0193] The obtained crystals had poor filterability, taking a long time to filter, and the wet material before drying was in a slurry state, with poor operability.

[0194] When measuring the particle size of the obtained crystals, the mode particle size was about 6 μm. The particle size distribution of the crystals obtained under Condition 1 was measured ( Figure 5-1 ).

[0195] Condition 2

[0196] To compound3 (17.58 g) of ethanol solution (53.20 g) was added with an aqueous sodium hydroxide solution (24%, 4.67 g) and purified water (16.20 g), and stirred at 40 °C for 3 hours to obtain a solution of compound 4 (73.14 g). To 18.28 g of the obtained solution of compound 4 , phosphoric acid (1.91 g, 2.6 equivalents) and purified water (7.49 g, 1.5 times the volume based on the weight of compound 4 ) were added at 35 °C, and after stirring for 1 hour, seed crystals (125 mg, 0.025 times the weight based on the weight of compound 4 ) were added. 17 hours and 30 minutes after the addition of the seed crystals, purified water (30.0 mL, 6.0 times the volume based on the weight of compound 4 ) was added over 2 hours. After 10 hours, the reaction temperature was set to 20 °C and stirring was continued. After 27 hours, the solid was filtered out and washed with purified water (25.23 g, 5 times the volume based on the weight of compound 4 ). The solid was dried at 50 °C to obtain the crystal of the present invention (4.01 g).

[0197] Sampling was carried out 16 hours and 21 hours after the addition of the seed crystals, and quantitative analysis was performed by HPLC to calculate the precipitation rate of the crystals, and the results were 4% and 97% respectively.

[0198] It can be seen that under Condition 2, when the amount of purified water before seeding is small, the crystal precipitation rate becomes extremely low. That is, it can be known that in order to precipitate the crystal of the present invention, the amount of purified water added to the ethanol solution, that is, the composition of the crystallization solvent, is important. In addition, when further purified water is added to the solution with a low precipitation rate, although crystals precipitate, as in Condition 1, crystals with poor operability such as filtration are obtained.

[0199] Condition 3

[0200] To a solution of compound 3 (26.37 g) in ethanol solution (79.82 g), an aqueous sodium hydroxide solution (24%, 7.01 g) and purified water (24.30 g) were added, and stirred at 40 °C for 4 hours and 30 minutes to obtain a solution of compound 4 . To the obtained solution of compound 4 , phosphoric acid (9.70 g, 2.2 equivalents) and purified water (45.00 g, 1.5 times the volume based on the weight of compound 4 ) were added at 20 °C, and further purified water (45.13 g, relative to compound 4In terms of the weight, it is 1.5 times the amount by volume), and after stirring at 30 °C for 40 minutes, seed crystals (751 mg, 0.025 times the amount by weight in terms of the weight of the compound 4 were added. After 16 hours, purified water (135 mL, 4.5 times the amount by volume in terms of the weight of the compound 4 ) was added over 2 hours. After 6 hours, the temperature was set to 20 °C and stirring was continued for 1 hour. The solid was filtered out and washed with purified water (150.02 g, 5 times the amount by volume in terms of the weight of the compound 4 ). The solid was dried at 50 °C to obtain the crystal of the present invention (26.95 g).

[0201] When measuring the particle size of the obtained crystal, the mode particle size was about 10 μm. The operability such as filtration of the obtained crystal was good. The particle size distribution of the crystal obtained under Condition 3 was measured ( Figure 5-2 ).

[0202] As described above, by setting the temperature to around 30 °C, adding seed crystals after adding purified water in the amount described in Condition 3, allowing the crystal to grow significantly by leaving it for a certain period of time, and then further adding purified water, crystals can be obtained with a good precipitation rate and operability.

[0203] [Table 8]

[0204] Table 8

[0205] Relationship between the amount of purified water added, the seeding temperature, and the precipitation rate under Experimental Conditions 2 and 3

[0206]

[0207] Reaction apparatus and stirring conditions

[0208] Condition 2

[0209] Reaction apparatus: EasyMax (registered trademark) (Mettler-Toledo Co., Ltd.)

[0210] Stirring conditions: 300 rpm

[0211] Condition 3

[0212] Reaction apparatus: OptyMax (registered trademark) (Mettler-Toledo Co., Ltd.)

[0213] Stirring conditions: 250 rpm

[0214] HPLC measurement conditions

[0215] Equipment name:

[0216] Column: GL Science, Inertsil ODS-3V (5 μm, 4.6 X 150 mm)

[0217] Mobile phase A: water / acetonitrile / trifluoroacetic acid = 1900:100:1

[0218] Mobile phase B: water / acetonitrile / trifluoroacetic acid = 100:1900:1

[0219] Particle size determination conditions

[0220] Equipment name: Particle Track (registered trademark) MALVERN, Mastersizer 2000 (wet type)

[0221] Measurement range: 0.020 - 2000.000 μm

[0222] Measurement time: 10 seconds

[0223] Measurement intensity range: 3.0 - 20.0%

[0224] Stirring speed: approximately 2000 rpm

[0225] As described above, like in Condition 3, after adding sufficient purified water, seeds are added, left for a certain period of time to allow the crystals to grow significantly, and then further purified water is added. Thereby, a sufficient precipitation amount can be ensured, and at the same time, crystals containing a large amount of crystals with good fluidity, filterability, and large chord length can be obtained. Thus, the operability such as filtration is improved.

[0226] Experimental Example 8 Determination of human MC1R agonist

[0227] Using the crystals of the present invention, according to the following method described in Experimental Example 1 of Patent Document 1, the intracellular cAMP concentration was measured to calculate the EC 50 value.

[0228] (1) Cell culture method

[0229] In the determination of human MC1R agonist activity, the human melanoma cell line HBL was used. Cultivation of HBL: It was cultivated using F-10 Nutrient Mixture containing 10% FCS, Penicillin-streptomycin.

[0230] (2) cAMP detection and data calculation

[0231] Mix the compound solutions at various concentrations with the cAMP detection buffer (HBSS (Hank's Balanced Salt Solution) containing 10 mM HEPES and 0.1% BSA), and dispense them into a 96-well plate. HBL was suspended in the cAMP detection buffer containing 0.5 mM IBMX at a concentration of 5×10 4 / mL, dispensed into the above 96-well plate and then mixed. After standing at 37°C for 30 minutes, the intracellular cAMP concentration was measured by fluorescence method using Envision (ex. 320 nm, em. 590 nm and 665 nm). For the obtained data, according to the ratio (measurement value at 665 nm / measurement value at 590 nm × 10000), the quantitative value of the cAMP concentration was calculated using Prism 5.02, and the induction% value (induction% value) was calculated (the average cAMP concentration of the background medium (vehicle) was set as 0%, and the average cAMP concentration of αMSH at 10 -6 M was set as 100% for each sample), and the EC 50 value was calculated.

[0232] As a result, the crystal of the present invention showed an EC 50 value of 5.3 nM and was a crystal having strong human MC1R agonist activity.

[0233] Industrial Applicability

[0234] Regarding the crystal of the present invention, the solvent used in obtaining the crystal does not remain, it has excellent thermal stability, little weight change with respect to humidity, is stable, does not deliquesce, has excellent chemical stability, and from the viewpoint of safety, it is also a crystal that does not contain compounds that may have an adverse effect on organisms. In addition, the crystal can be obtained with good reproducibility by an industrially suitable method, and thus is an excellent crystal as a pharmaceutical raw material.

Claims

1. A crystal comprising equimolar amounts of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid.

2. The crystal according to claim 1, which is formed from equimolar amounts of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid.

3. The crystal according to any one of claims 1 or 2, which is a cocrystal of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid and phosphoric acid.

4. The crystal according to any one of claims 1 to 3, wherein, in the powder X-ray diffraction spectrum, peaks are shown at diffraction angles represented by 2θ of 5.7°, 11.5°, 13.9°, 19.0° and 21.9° (±0.2° respectively).

5. The crystal according to any one of claims 1 to 4, wherein, in the differential scanning calorimetry analysis, an endothermic peak is present at 230 °C to 240 °C.

6. The crystal according to any one of claims 1 to 5, which is obtained by adding a seed crystal to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid and a good solvent.

7. The crystal according to any one of claims 1 to 6, which is obtained by the following method: adding a poor solvent to a mixture of 1-{2-[(3S,4R)-1-[(3R,4R)-1-cyclopentyl-3-fluoro-4-(4-methoxyphenyl)pyrrolidine-3-carbonyl]-4-(methoxymethyl)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid and a good solvent, adding a seed crystal, and then further adding a poor solvent.

8. A melanocortin receptor 1 agonist containing the crystal according to any one of claims 1 to 7 as an active ingredient.

9. A pharmaceutical composition containing the crystal according to any one of claims 1 to 7 and a pharmaceutically acceptable additive.

10. The pharmaceutical composition according to claim 9, which is used for preventing or treating a disease whose condition is expected to be improved by activation of melanocortin receptor 1.

11. The pharmaceutical composition according to claim 10, wherein, The disease is one or more diseases selected from rheumatoid arthritis, gouty arthritis, osteoarthrosis, inflammatory bowel disease, systemic scleroderma, psoriasis, fibrosis, protoporphyria, systemic lupus erythematosus, melanoma, skin cancer, vitiligo, alopecia, pain, ischemia / reperfusion injury, inflammatory diseases of the brain, hepatitis, sepsis / septic shock, nephritis, transplantation, exacerbation of HIV disease, vasculitis, uveitis, retinitis pigmentosa, age-related macular degeneration, microbial infection, celiac disease, nephrotic syndrome, and melanoma infiltration.

12. A method for preventing or treating a disease whose symptoms are expected to be improved by activation of melanocortin receptor 1, which comprises administering to a patient an effective amount of the crystal according to any one of claims 1 to 7.

13. Use of the crystal according to any one of claims 1 to 7 in the manufacture of a medicament for preventing or treating a disease whose symptoms are expected to be improved by activation of melanocortin receptor 1.

14. The crystal according to any one of claims 1 to 7, which is used for preventing or treating a disease whose symptoms are expected to be improved by activation of melanocortin receptor 1.

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