Crystal of pyrrolidine compound

Abstract

To provide a crystal of pyrrolidine compound A having a certain quality that can be used as a drug substance.SOLUTION: The present invention provides a crystal 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 having a certain quality that can be used as a drug substance. Specifically, the present invention provides a crystal comprising an equimolar amount 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.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a crystal containing 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") and phosphoric acid. More specifically, the present invention relates to a crystal containing 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 active ingredient, and a pharmaceutical composition containing the same as an active ingredient. [Background technology]

[0002] International Publication WO2015 / 182723 pamphlet (hereinafter referred to as Patent Document 1) discloses that pyrrolidine compounds having melanocortin receptor 1 (MC1R) agonistic activity (agonist activity) or pharmacologically acceptable salts thereof, as well as these compounds and pharmaceutical compositions containing these compounds as active ingredients, are useful for the treatment or prevention of various diseases whose pathological conditions are expected to be improved by activation of MC1R, and describes the hydrochloride salt of pyrrolidine compound A in Example 19. However, Patent Document 1 does not describe or suggest anything about crystals of pyrrolidine compound A. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication WO2015 / 182723 Brochure Summary of the Invention [Problem to be solved by the invention]

[0004] An object of the present invention is to provide a crystal of pyrrolidine compound A having a certain quality that allows it to be used as a pharmaceutical active ingredient. [Means for solving the problem]

[0005] It was found that the hydrochloride of pyrrolidine compound A does not crystallize, has deliquescence, and is not suitable as a drug substance. Therefore, the present inventors have attempted crystallization of pyrrolidine compound A under more than 1000 conditions in order to obtain crystals having a certain quality that can be used as a drug substance. As a result, it was found that a crystal containing equimolar amounts of pyrrolidine compound A and phosphoric acid is a crystal having a certain quality that can be used as a drug substance from the viewpoints of purity, thermal stability, hygroscopicity, deliquescence, chemical stability, and safety, and thus the present invention was completed. Furthermore, it was found that the crystals containing the equimolar amounts of pyrrolidine compound A and phosphoric acid were difficult to crystallize. Therefore, the present inventors investigated conditions for obtaining crystals having sufficient purity with good reproducibility in a short time. Problems such as an increase in impurities, delayed precipitation, and deterioration of stirring fluidity and filterability due to fine crystals were observed depending on the crystallization temperature and the composition of the crystallization solvent. In order to solve these problems, the present inventors have intensively investigated the types, amounts, and ratios of reagents and solvents used for crystallization, as well as the crystallization procedures, and have found a method for efficiently obtaining crystals of a quality suitable for pharmaceutical raw materials that have few impurities and good operability such as filtration.

[0006] The present invention relates to the following: [1] An equimolar amount 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 Crystals containing lysine-4-carboxylate and phosphate. [2] The crystal according to [1] is composed of 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 either [1] or [2], which is a co-crystal 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 [1] to [3], which exhibits peaks at diffraction angles 2θ of 5.7°, 11.5°, 13.9°, 19.0° and 21.9° (each ±0.2°) in a powder X-ray diffraction spectrum. [5] The crystal according to any one of [1] to [4], which has an endothermic peak at 230°C to 240°C in differential scanning calorimetry. [6] 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)pyrrolidin-3-yl]-5-(trifluoromethyl)phenyl}piperidine-4-carboxylic acid, phosphoric acid, and a good solvent. [7] The crystal according to any one of [1] to [6] can be obtained by 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 comprising the crystal according to any one of [1] to [7] as an active ingredient. [9] A pharmaceutical composition comprising the crystal according to any one of [1] to [7] and a pharma- ceutical acceptable excipient.

[10] The pharmaceutical composition described in [9] for preventing or treating a disease whose pathological condition is expected to be improved by activation of melanocortin receptor 1.

[11] The disease is selected from rheumatoid arthritis, gouty arthritis, osteoarthritis, inflammatory bowel disease, systemic sclerosis, psoriasis, fibrosis, protoporphyria, systemic lupus erythematosus, melanoma, skin cancer, vitiligo, hair loss, 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. The pharmaceutical composition according to

[10] , for treating the above diseases.

[12] A method for preventing or treating a disease whose pathology is expected to be improved by activation of melanocortin receptor 1, comprising administering to a patient an effective amount of a crystal described in any one of [1] to [7].

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

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

[0007] The present invention also relates to the following:

[15] A method for producing the crystal according to any one of [1] to [5], comprising 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.

[16] The production method according to

[15] , which comprises 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 seed crystals, and then further adding the poor solvent. Effect of the Invention

[0008] The crystals containing equimolar amounts of pyrrolidine compound A and phosphoric acid are free of residual solvent used in obtaining the crystals, have excellent thermal stability, are stable with little weight change against humidity, do not deliquesce, have excellent chemical stability, and do not contain compounds that may have adverse effects on living bodies from a safety perspective. The crystals can be obtained reproducibly by an industrially suitable method, and therefore are useful as pharmaceutical ingredients. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows the results of powder X-ray diffraction measurement of the crystals of the present invention. [Diagram 2] FIG. 2 is a diagram showing the results of differential scanning calorimetry of the crystal of the present invention. [Diagram 3] FIG. 3 is a diagram showing an Orthopaedic diagram based on single crystal X-ray diffraction measurement of the molecules in the crystal of the present invention. [Figure 4] FIG. 4 is a diagram showing a packing diagram (a-axis projection diagram) based on single crystal X-ray diffraction measurement of the molecules in the crystal of the present invention. [Figure 5-1] FIG. 5-1 is a graph showing the particle size distribution of the crystals obtained under condition 1 in Experimental Example 7. [Figure 5-2] FIG. 5-2 is a graph showing the particle size distribution of the crystals obtained under condition 3 in Experimental Example 7. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The present invention relates to a crystal containing equimolar amounts of pyrrolidine compound A represented by the following formula and phosphoric acid, and a pharmaceutical composition containing the same as an active ingredient. [ka]

[0011] The pyrrolidine compound A and / or phosphoric acid in the crystal of the present invention may be isotope-doped (e.g., 3 H, 13 C. 14 C. 15 N, 18 F, 32 This includes compounds labeled with fluorine (e.g., P) and deuterium conversions.

[0012] In the present invention, the crystals containing equimolar amounts of pyrrolidine compound A and phosphoric acid are free of other molecules such as the solvent used in obtaining the crystals, and the crystals are constituted by pyrrolidine compound A and phosphoric acid in a molar ratio of 1:1, i.e., 1 molar equivalent of phosphoric acid is contained relative to pyrrolidine compound A.

[0013] A preferred embodiment of the crystal of the present invention is a cocrystal in which pyrrolidine compound A and phosphoric acid do not form a salt, but are bound to each other by a nonionic bond and / or a noncovalent bond at a molar ratio of 1:1. In one embodiment, the crystal of the present invention has a powder X-ray diffraction pattern as shown in FIG. 1, and characteristic peaks include 5.7°, 11.5°, 13.9°, 19.0°, and / or 21.9° (each ±0.2°) in terms of diffraction angle indicated by 2θ, and more specifically, 5.7°, 11.5°, 13.9°, 17.4°, 19.0°, 20.4°, and / or 21.9° (each ±0.2°) in terms of diffraction angle indicated by 2θ. ), more specifically, at diffraction angles of 5.7°, 7.4°, 11.5°, 12.3°, 13.9°, 17.4°, 19.0°, 20.4°, and / or 21.9° (each ±0.2°) at 2θ, and particularly, at diffraction angles of 2θ, the peaks shown in Table 2 below (hereinafter, crystals having these peaks may be referred to as B-type crystals or B-type phosphate crystals). In another embodiment, the B-type crystals show the differential scanning calorimetry (hereinafter, may be referred to as DSC) curve shown in FIG. 2, and have an endothermic peak at 230°C to 240°C.

[0014] The crystals of the present invention have a residual solvent that satisfies the requirements of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (hereinafter referred to as "the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use"). The crystals are referred to as ICH.) It has an advantageous effect in that the crystals are below the standard values ​​set by the ICH guidelines. Another advantage is that the crystals are below the standard values ​​set by the ICH guidelines for organic impurities, inorganic impurities, residual metals, residual solvents, genotoxic impurities, etc.

[0015] The crystals of the present invention are preferably produced as crystals having a mode diameter of non-aggregated crystals of 7 μm or more, more preferably 7 to 15 μm, and particularly preferably 8 to 12 μm. The mode diameter is the particle diameter when the volume percentage of the particle size distribution is at a maximum value. In the method for producing crystals of the present invention, by making at least 70% or more, preferably 80% or more, and more preferably 90% or more of the total crystals have such a mode diameter, the operability, such as the ease of filtering the crystals, is improved.

[0016] The crystals of the present invention can be obtained by reacting 1 to 10 moles, preferably 1 to 5 moles, of phosphoric acid with 1 mole of pyrrolidine compound A, which can be produced, for example, in accordance with the method described in Example 19 of Patent Document 1. In addition, the crystals can also be produced by the method shown in the Examples described later.

[0017] The solvent used to obtain the crystal of the present invention can be appropriately selected, and for example, a good solvent or a poor solvent can be used alone or in appropriate combination. The good solvent is not limited as long as it is a solvent in which the pyrrolidine compound A has a high solubility, and examples thereof include ketones (e.g., acetone, 2-butanone, etc.), esters (e.g., ethyl acetate, methyl acetate, etc.), alcohols (e.g., methanol, ethanol, i-propanol, etc.), and mixtures of these solvents. The poor solvent is not limited as long as it is a solvent in which the pyrrolidine compound A has a low solubility, and examples thereof include 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.

[0018] As one embodiment of the method for obtaining the crystal of the present invention, phosphoric acid is added to a mixture of pyrrolidine compound A and a good solvent, seed crystals are added to the obtained mixture, and the mixture is filtered. A preferred method is to dissolve pyrrolidine compound A in a good solvent, add phosphoric acid, add seed crystals to the obtained mixture, and then filter. A more preferred method is to dissolve pyrrolidine compound A in ethyl acetate or ethanol, add phosphoric acid, add seed crystals to the obtained mixture, and then filter.

[0019] In another embodiment, phosphoric acid is added to a mixture of pyrrolidine compound A and a good solvent, a poor solvent is added to the resulting mixture, and the mixture is filtered. Preferably, 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 the mixture is filtered. More preferably, 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, a poor solvent is further added, and the mixture is filtered. Even more preferably, 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 water is further added, and the mixture is filtered.

[0020] Examples of suitable combinations and ratios of a good solvent and a poor solvent when used in combination include ethanol:toluene=1:9, ethanol:diisopropyl ether=3:7, and acetone:toluene=3:7. Examples of suitable good solvents when used in combination with water include ethyl acetate and ethanol. When a good solvent and a poor solvent, specifically water, are used in combination, and water is added twice before and after the addition of seed crystals, the amount of water added before the addition of seed crystals (first water addition) is preferably 3 times the weight of pyrrolidine compound A in volume ratio, and the amount of water added after the addition of seed crystals (second water addition) is preferably 4.5 times the weight of pyrrolidine compound A in volume ratio. In another embodiment, the amount of water added is 5 to 10 times the weight of pyrrolidine compound A in volume ratio, and is added in portions before and after the addition of seed crystals. More preferably, the amount of water is 6 to 9 times the weight of pyrrolidine compound A in volume ratio, and more preferably 7.5 times. In another embodiment, the volume ratio of the amount of water added in the first step to the amount of water added in the second step is 1:1 to 1:2, preferably A preferred example is 2:3. Another embodiment is to add water before the addition of seed crystals to such an extent that 70% or more, preferably 80% or more, of the crystals have precipitated before the addition of water after the addition of seed crystals. The temperature at which the seed crystals are added is set to 28 to 32°C, preferably 30°C. It is also preferred to select such a temperature that 70% or more, preferably 80% or more, of the crystals have precipitated before the addition of water after the addition of seed crystals.

[0021] The seed crystals of the present invention can be obtained by the methods described later in Example 3, Experimental Example 2, or Experimental Example 3. The seed crystals of the present invention can also be obtained by using the crystals obtained by these methods as seed crystals of the present invention, for example, by the methods described in Examples 1 or 2 or methods similar to the methods described in Examples 1 or 2.

[0022] Since the crystal of the present invention has human MC1R agonist activity, it can be used as an active ingredient of a melanocortin receptor 1 agonist, and the crystal of the present invention and a pharmaceutical composition containing the crystal as an active ingredient are useful for treating or preventing various diseases whose pathology is expected to be improved by activating MC1R. Examples of such diseases include one or more diseases selected from rheumatoid arthritis, gouty arthritis, osteoarthritis, inflammatory bowel disease, systemic sclerosis, psoriasis, fibrosis, protoporphyria (e.g., erythroblastic protoporphyria, etc.), systemic lupus erythematosus, melanoma, skin cancer, vitiligo, hair loss, pain, ischemia / reperfusion injury, brain inflammatory disease, 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. In particular, it is useful for the treatment or prevention of one or more diseases selected from systemic sclerosis, psoriasis, protoporphyria, melanoma, skin cancer, vitiligo, alopecia, retinitis pigmentosa, age-related macular degeneration, and nephrotic syndrome, etc. In particular, it is useful for the treatment or prevention of one or more diseases selected from systemic sclerosis, protoporphyria, melanoma, vitiligo, retinitis pigmentosa, age-related macular degeneration, and nephrotic syndrome, etc.

[0023] A 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 pharma- ceutical acceptable additives, such as excipients, disintegrants, binders, lubricants, coating agents, dyes, diluents, bases, and isotonicity agents.

[0024] The crystal of the present invention and the pharmaceutical composition containing it as an active ingredient can be prepared into an appropriate dosage form, such as a powder, injection, tablet, capsule, topical preparation, etc., and then administered to a patient by an appropriate administration method according to the dosage form, such as intravenous administration, oral administration, transdermal administration, etc. In the present invention, the term "patient" refers to an individual who is the target of prevention or treatment by the crystal of the present invention, and is preferably a mammal, more preferably a human.

[0025] The dosage is determined in consideration of the age, weight, general health condition, sex, diet, administration time, administration method, excretion rate, drug combination, and the degree of the disease of the patient being treated at the time of administration, or other factors. The crystal of the present invention and the pharmaceutical composition containing it as an active ingredient are low toxic and can be used safely, and the daily dosage (i.e., effective amount) varies depending on the condition and weight of the patient, administration route, etc., but for example, it is preferably administered parenterally at about 0.0001 to 1000 mg / person / day, preferably about 0.001 to 1000 mg / person / day, particularly preferably 0.01 to 500 mg / person / day, and orally at about 0.0001 to 1000 mg / person / day, preferably 0.01 to 500 mg / person / day.

[0026] In the present invention, "prevention" refers to the act of administering the crystal of the present invention or a pharmaceutical composition containing the same to an individual who does not have a disease, disorder, or symptom. It means administering the crystal of the present invention or a pharmaceutical composition containing the same to an individual who has already developed a disease, disorder, or symptom. Therefore, administering to an individual who has already developed a disease, disorder, or symptom in order to prevent the deterioration of symptoms, attacks, or recurrence is one aspect of "treatment". EXAMPLES

[0027] The present invention will be described in detail below with reference to examples and experimental examples, but the present invention is not limited thereto. In addition, "equivalent" means "molar equivalent".

[0028] Example 1 Synthesis of the crystal of the present invention [ka] compound 1 (26.18 g) was dissolved in dichloromethane (207 mL) and the compound 2 (4.4 mL) and acetic acid (3.56 mL) were added and stirred at room temperature for 30 minutes. Sodium triacetoxyborohydride (13.2 g) was then added and stirred at room temperature for 1 hour. A saturated aqueous solution of sodium bicarbonate was added and stirred, and then extracted with dichloromethane. The organic layer obtained was washed with saturated saline, 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) and then silica gel column chromatography (chloroform:methanol=100:0 to 95:5) to obtain compound 1. 3 (24.25 g) was obtained as a colorless powder (MS (ESI): m / z 690 [M+H] + ).

[0029] compound 3(24.24 g) was dissolved in methanol (240 mL), and an aqueous sodium hydroxide solution (2 mol / L, 70.2 mL) was added thereto and stirred at room temperature for 19 hours. After that, hydrochloric acid water (2 mol / L, 74 mL) was further added, and the reaction solution was concentrated under reduced pressure. Water and ethyl acetate were added to the concentrated residue, and the mixture was stirred and extracted with ethyl acetate. The obtained organic layer was washed with phosphate buffer solution (0.1 mol / L, 300 mL) and saline, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform:methanol = 100:0 to 90:10), dissolved in ethyl acetate, and phosphate buffer solution (0.1 mol / L, 200 mL) was added thereto and stirred at room temperature, and extracted with ethyl acetate. The organic layer was washed with water and saturated saline, dried over magnesium sulfate, and concentrated under reduced pressure to obtain compound 1. 4 (23.7 g) was obtained as a colorless powder (MS (ESI): m / z 676 [M+H] + ).

[0030] compound 4 (135 mg) was dissolved in ethanol (0.7 mL), and a small amount of the crystals of the present invention was added as seed crystals. A phosphoric acid solution prepared by dissolving phosphoric acid (25 mg) in ethanol (0.5 mL) was added thereto, and ethanol (0.2 mL) was further added and stirred at room temperature overnight. The precipitated crystals were collected by filtration, washed with ethyl acetate (0.6 mL), and then dried under reduced pressure at 50°C for 4 hours to obtain 112.7 mg of the crystals of the present invention. The presence or absence of residual solvent was confirmed by 1H-NMR, but no residual solvent was found. The elemental analysis measurement results are shown in Table 1 below.

[0031] [Table 1]

[0032] <X-ray powder diffraction (hereinafter sometimes referred to as XRPD) measurement> The measurements were carried out using a powder X-ray diffraction measuring device X'PertPro (manufactured by PANalytical B.V.) under the following conditions. X-ray generator: X-ray tube (anticathode: copper, tube voltage: 45 kV, tube current: 40 mA) Incident optical system: focusing mirror Receiving optical system: High-speed semiconductor array detector (X-Celerator), extended receiving arm Sample stage: HTS sample stage (oscillates in the X-axis direction with a width of 4 mm) Number of measurements: 5 (angle of incidence changed to -2, -1, 0, 1, and 2°) Measurement range: 2θ=3~40° Scan speed: 0.668451° / sec Step: 0.0167°

[0033] The results are shown in Figure 1. When the peak intensity at a diffraction angle of 5.7°, indicated by 2θ, is taken as 100, the peaks having a relative peak intensity of 5 or more are as shown in Table 2 below.

[0034] [Table 2]

[0035] <Differential scanning calorimetry (DSC) measurement> Measurements were performed under the following conditions using a differential scanning calorimeter X-DSC7000 (SII NanoTechnology Co., Ltd.). Heating rate: 10℃ / min (25℃~300℃) Atmosphere: Nitrogen 100mL / min The results are shown in Figure 2. An endothermic peak was observed at approximately 230°C to 240°C.

[0036] <Single crystal X-ray diffraction measurement> Add about half a small spatula of the present invention to 2 mL of ethanol and dissolve it. The crystals were left to stand for days to allow crystallization. The obtained crystals were analyzed by determining the lattice constants and measuring the diffraction peak intensities at 23°C using a single crystal X-ray diffractometer R-AXIS RAPID / R (Rigaku Corporation) (CuKα radiation), followed by phase determination by a direct method and refinement of the structure by the full-matrix least squares method. The obtained crystallographic data and crystal structure analysis results are shown in Table 3. The reliability factor (R value) was 3.06%, and other various parameters also showed that the results of this crystal structure analysis were sufficiently reliable.

[0037] [Table 3]

[0038] The Orthopaedic diagram of the molecules in the crystal of the present invention is shown in Figure 3, and the packing diagram is shown in Figure 4. In the crystal, one molecule each of pyrrolidine compound A and phosphoric acid was present independently in the asymmetric unit.

[0039] As a result of verifying the absolute configuration of the crystal of the present invention, the Flack parameter was 0.02 (3), and therefore it was confirmed that the crystal of the present invention is a crystal having one molecule 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 one molecule of phosphoric acid as constituent units as shown in FIG. 3.

[0040] The guidance issued by the U.S. Food and Drug Administration (FDA) (Regulatory Classification of Pharmaceutical Co-Crystals Guidance for Industry) states that co-crystals are classified into crystal lattices. It is described as a crystal in which two or more different molecules are contained within the crystal, and they are bound by nonionic and / or noncovalent bonds. Since the absence of a covalent bond between pyrrolidine compound A and phosphate has already been confirmed by the above-mentioned crystal structure analysis, we examined whether or not an ionic interaction is observed between pyrrolidine compound A and phosphate.

[0041] Since phosphoric acid is a Bronsted acid, when an ionic interaction with pyrrolidine compound A is observed, pyrrolidine compound A must accept a proton as a Bronsted base. In the case of pyrrolidine compound A, there are three nitrogens (N18, N19, and N20) that can accept a proton from phosphoric acid. Here, all hydrogen bonds in the crystal of the present invention The binding sites are shown in Table 4.

[0042] [Table 4]

[0043] (1) N18 N18 is an sp3 hybridized nitrogen, and is the most basic nitrogen in pyrrolidine compound A, which may accept a proton from phosphate. However, 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 it was determined that a proton was donated. Therefore, N18 cannot accept a proton from phosphate. At the same time, since the interatomic distance between N18 and any oxygen of the adjacent phosphate exceeds the sum of the van der Waals radii, it was determined that no phosphate exists within a distance that allows ionic interaction.

[0044] (2) N19 Since N19 is an sp2 hybridized nitrogen, it is considered to be weakly basic and unable to form a salt. At the same time, because the interatomic distance between N19 and the closest oxygen atom (O13) of the phosphate is the sum of the van der Waals radii, it was determined that the distance is not close enough for ionic interaction.

[0045] (3) N20 Since N20 is also an sp2 hybridized nitrogen, it is considered to be weakly basic and unable to form a salt. At the same time, since the interatomic distance between N20 and any oxygen atom of the adjacent phosphate exceeds the sum of the van der Waals radii, it was determined that no phosphate exists within a distance that would allow ionic interaction.

[0046] From the above results, it was clarified that the bond between the nitrogen (N18, N19, N20) of pyrrolidine compound A and phosphoric acid in the crystal of the present invention is not due to ionic interaction, pyrrolidine compound A and phosphoric acid do not form a salt, and it was determined that the crystal of the present invention is a cocrystal. The crystal obtained in this example is referred to as type B crystal in this specification.

[0047] Example 2 Synthesis of the crystal of the present invention (2) [ka] compound 3 A solution of (276 mg) in ethanol (1.4 mL) was added to 1,2-ethane disulfone The acid hydrate (38 mg) was added and stirred at room temperature for 40 minutes. The crystals were collected by filtration and washed twice with ethanol (0.84 mL). The crystals were dried at 40° C. or less to obtain compound (III). 5 (178mg ) was obtained. 5To a suspension of 37.1 kg of ethyl acetate (167.5 kg), a solution of potassium carbonate (6.5 kg) in water (148.3 L) and water (36.8 L) were added at room temperature in that order, and 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 off. After concentrating 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. The mixture was diluted with ethanol (44 kg), and 24% aqueous sodium hydroxide solution (8.7 kg) and water (30.1 kg) were added at room temperature in that order, and the mixture was stirred at 40° C. for 5 hours to obtain compound (1,2-dichlorophenyl)-1,2-dichlorophenyl ... 4 The compound was obtained. 4 To the solution, a solution of phosphoric acid (12.0 kg) in water (55.7 L) and water (55.7 L) were added in this order at 30°C. The crystals of the present invention were added as seed crystals (928 g) and stirred for 14 hours. Water (167.0 L) was added, stirred for 4 hours, and then cooled to 25°C. The solid was collected by filtration and washed with water (182 L). The solid was dried at 50°C or less to obtain the crystals of the present invention (35.2 kg). These crystals had excellent operability, such as filterability.

[0048] Experimental Example 1 Crystallization study of pyrrolidine compound A Using pyrrolidine compound A, crystallization studies were carried out using 25 single solvents and 44 mixed solvents by long-term storage, 96 mixed solvents prepared by changing the mixing ratio of the solvents using 2 anti-solvents and 12 good solvents by one month of stirring, and 26 added solvents by grinding. As a result, crystals were obtained by long-term storage and one month of stirring. Powder X-ray diffraction of the obtained crystals showed that the peaks of all crystals were the same and they had the same crystal form. Crystallization conditions were examined, but solvent remained in the crystals under all conditions under which crystals were obtained. A process for drying the crystals was also examined, but it was not possible to reduce the residual solvent to below the residual solvent standard value set by ICH. Therefore, although the free pyrrolidine compound A crystallizes, the solvent used to obtain the crystals remains, and the compound has low chemical stability, so that the compound cannot be used as a pharmaceutical ingredient.

[0049] Experimental Example 2 Crystallization study of mixture containing pyrrolidine compound A (1) About 900 mg of pyrrolidine compound A was dissolved in 30 mL of tetrahydrofuran, and 100 μL was dispensed into each vial of a 96-well plate (about 3 mg / vial). In addition, 21 kinds of acids including phosphoric acid, hydrochloric acid, L(-)-malic acid, L(+)-tartaric acid, maleic acid, sulfuric acid, and malonic acid, and 8 kinds of bases including sodium hydroxide and L-arginine (hereinafter sometimes referred to as counter compounds) were dissolved in 8 kinds of solvents described below, and 45 μL of 0.1 mol / L solution (90 μL of 0.05 mol / L solution for some acids) was dispensed into each vial. After evaporating the solvent by leaving it open for 1 day and night, it was dried under reduced pressure for 4 hours. After dispensing 250 μL of 8 kinds of solvents including ethyl acetate, acetone, and toluene into each vial, it was stirred by ultrasonic for 5 minutes, sealed, and stirred at room temperature for 6 days. For vials with precipitates, the precipitates were collected by filtration, and XRPD was measured. For vials without precipitate, the solvent was evaporated at room temperature, and if a solid was observed, it was filtered off and subjected to XRPD measurement. The XRPD measurement device and measurement conditions were the same as those in Example 1.

[0050] Phosphoric acid type A crystals, L(-)-malic acid type D crystals, L(+)-tartaric acid type E crystals, and maleic acid type F crystals were obtained from the vials containing phosphoric acid and ethyl acetate, L(-)-malic acid and toluene, L(+)-tartaric acid and acetone, and maleic acid and toluene, respectively. In addition to these, crystals of only the counter compound, i.e., There were some vials in which crystals not containing pyrrolidine compound A were observed. On the other hand, no crystals were obtained from the vials containing hydrochloric acid in any of the eight solvents. Furthermore, no crystals were obtained from the vials containing sulfuric acid, malonic acid, or L-arginine in any of the solvents.

[0051] Experimental Example 3 Crystallization study of mixture containing pyrrolidine compound A (2) About 800 mg of pyrrolidine compound A was dissolved in 40 mL of tetrahydrofuran, and 100 μL was dispensed into each vial of a 96-well plate (about 2 mg / vial). In addition, 22 kinds of acids including phosphoric acid, hydrochloric acid, L(-)-malic acid, L(+)-tartaric acid, maleic acid, sulfuric acid and malonic acid, and 8 kinds of bases including sodium hydroxide and L-arginine (hereinafter sometimes referred to as counter compounds) were dissolved in 12 kinds of solvents described below, and 30 μL of 0.1 mol / L solution (60 μL of 0.05 mol / L solution for some acids) was dispensed into each vial. After dispensing, nitrogen was sprayed to evaporate the solvent. 200 μL of 12 kinds of 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 were dispensed into each vial, sealed, and stirred at room temperature for 3 days. For vials with precipitates, the precipitates were filtered and XRPD was measured. For vials without precipitates, the solvent was evaporated at room temperature, and if a solid was observed after about one month, the solid was collected and XRPD was measured. The XRPD measurement device and measurement conditions were the same as those in Example 1.

[0052] The vials from which crystals were obtained are shown in Table 5 below. In the table, - indicates that no crystals were obtained, and A and B indicate that A-type crystals and B-type crystals were obtained, respectively. That is, the maleic acid F-type crystals obtained this time matched the maleic acid F-type crystals obtained in Experimental Example 2 in XRPD pattern, and the phosphoric acid B-type crystals obtained this time were different from the phosphoric acid A-type crystals obtained in Experimental Example 2. As in Experimental Example 2, there were also vials in which crystals of only the counter compound were confirmed. On the other hand, no crystals were obtained from the vials to which hydrochloric acid was added in all 12 solvents. In addition, for L(-)-malic acid and L(+)-tartaric acid, crystals were obtained in Experimental Example 2, but no crystals were obtained in all 12 solvents this time.

[0053]

Table 5

[0054] Experimental Example 4 Examination of Crystallization of Mixture Containing Pyrrolidine Compound A (3) Among the crystals obtained in Experimental Examples 2 and 3, except for the A-form phosphate crystals and B-form phosphate crystals, the reproducibility was confirmed. As a result, the D-form crystals of L(-)-malic acid, the E-form crystals of L(+)-tartaric acid, the J-form sulfate crystals, and the K-form crystals of L-arginine could not be obtained again. On the other hand, crystals could be obtained again in the combination with maleic acid and malonic acid.

[0055] <L(-)-Malic Acid, L(+)-Tartaric Acid, and Sulfuric Acid> Approximately 80 mg of pyrrolidine compound A was dissolved in 4 mL of tetrahydrofuran, and 100 μL each was dispensed into each vial of a 96-well plate (about 2 mg / vial). Also, 30 μL of a 0.1 mol / L solution of L(-)-malic acid, L(+)-tartaric acid, and sulfuric acid dissolved in the solvents described below was dispensed into each vial. After dispensing, the solvent was evaporated by nitrogen blowing. Then, 200 μL of toluene was dispensed into the vial containing L(-)-malic acid, 200 μL of acetone was dispensed into the vial containing L(+)-tartaric acid, and 200 μL of a mixed solvent of ethyl acetate and heptane with a mixing ratio of 3:7 was dispensed into the vial containing sulfuric acid. Eight sealed samples were prepared for each sample. A total of 24 vials were stirred at room temperature for 7 days, but no precipitate was obtained in any of the vials.

[0056] <L-Arginine> Approximately 65 mg of pyrrolidine compound A was weighed out and dissolved in 0.3 mL of ethanol and 2.1 mL of toluene at room temperature. Approximately 19 mg of L-arginine was weighed out and dissolved in 0.6 mL of ethanol and 0.6 mL of water, and added to the solution of pyrrolidine compound A. Since it was a solution, the solvent was evaporated by blowing nitrogen, and a candy-like substance containing some white powder was obtained. This was redissolved in 0.3 mL of ethanol, and 0.6 mL of diisopropyl ether was added dropwise. The L-arginine K-type crystal obtained in Experimental Example 3 was added as a seed crystal, but it dissolved, so 1.2 mL of toluene was added and stirred for a day and night. Since it was a solution, the solvent was evaporated by blowing nitrogen, and the residue was measured by XRPD, and it was amorphous. 0.6 mL of heptane was added to this, and it was stirred at room temperature overnight, and then observed under a microscope. No crystalline components were observed.

[0057] <Maleic acid F crystals> About 65 mg of pyrrolidine compound A was weighed out and dissolved in 0.5 mL of toluene at room temperature. When about 13 mg of maleic acid was dissolved in 75 μL of ethanol and a small amount of the previously obtained maleic acid F type crystals were added as seed crystals, the reaction solution became gel-like (agar-like) and could not be stirred. When 1.0 mL of toluene was further added and the mixture was rubbed vigorously with a spatula, a suspension was formed. The filtered crystals were dried under reduced pressure at 40°C for 5 hours to obtain 57 mg of maleic acid F type crystals. It was confirmed by 1H-NMR that 0.1 equivalent of toluene remained in the obtained crystals.

[0058] <Malonic acid G crystals> Approximately 325 mg of pyrrolidine compound A was weighed out and dissolved in 1 mL of acetone at room temperature. 55 mg of malonic acid was weighed out and dissolved in 0.5 mL of acetone at room temperature, and added to the solution of pyrrolidine compound A. 6 mL of toluene was added dropwise to this solution. A small amount of malonic acid type I crystals described below was added as seed crystals, and the mixture was stirred at room temperature for two nights. The entire amount was filtered and dried under reduced pressure at 40°C for 3.5 hours to obtain 317 mg of crystals. It was confirmed by 1H-NMR that the obtained crystals contained 1 equivalent of toluene. In order to replace toluene with water, a dynamic vapor sorption measurement (DVS) device was used under conditions of 25°C and 70% RH. After storage for 72 hours, amorphization was observed.

[0059] <Malonic acid H-type crystals> Approximately 65 mg of pyrrolidine compound A was weighed out and dissolved in 2 mL of a mixed solvent of ethyl acetate and heptane in a mixing ratio of 3:7 at room temperature. 11 mg of malonic acid was weighed out and dissolved in 0.3 mL of ethyl acetate at room temperature, and added to the solution of pyrrolidine compound A. A hard gum-like substance was formed, so 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, 3 mL of ethyl acetate was added to the dried product, and the mixture was stirred, and 1.2 mL of heptane was gradually added. When a small amount of malonic acid G-type crystals obtained in Experimental Example 3 was added as seed crystals, a precipitate was rapidly formed. After stirring at room temperature for 2 nights, the entire amount was filtered and dried under reduced pressure at 40°C for 3.5 hours, and 38 mg of crystals was obtained.

[0060] <Malonic acid type I crystal> Approximately 65 mg of pyrrolidine compound A was weighed out and dissolved in 2.3 mL of a mixed solvent of acetone and toluene in a mixing ratio of 1:9 at room temperature. 11 mg of malonic acid was weighed out and dissolved in 0.1 mL of acetone at room temperature, and added to the solution of pyrrolidine compound A. The mixture was stirred at room temperature for three nights, filtered, and dried under reduced pressure at 40°C for two hours to obtain 51 mg of crystals. It was confirmed by 1H-NMR that the obtained crystals contained 1 equivalent of toluene. Since this crystal may be a toluene solvate, in order to confirm whether a solvent-free crystal can be obtained by further drying under reduced pressure, approximately 5 mg was dried under reduced pressure at 60°C for four hours, and it was confirmed that approximately 0.5 equivalents of toluene remained and that there was no change in the crystal form.

[0061] Example 3 Synthesis of the crystal of the present invention (3) Pyrrolidine compound A and phosphoric acid in the amounts shown in Table 6 below were added to the solvent shown below and stirred for the time shown below. The results are shown in Table 6 below. The XRPD measurement device and measurement conditions were the same as those in Example 1. The XRPD results show that A is the same crystal as the phosphoric acid type A crystal obtained in Experimental Example 2, and B is the same crystal as the phosphoric acid type B crystal obtained in Experimental Example 3. Furthermore, the phosphoric acid type B crystal was the same as the crystal of the present invention obtained in Example 1.

[0062] [Table 6]

[0063] Method (a) yielded a mixture of type A crystals and type B crystals. Elemental analysis of these crystals revealed that they contained 1.5 equivalents of phosphoric acid. Method (f) using an acetone / toluene solvent yielded new type C crystals. Elemental analysis of these type C crystals revealed that they contained 2 equivalents of phosphoric acid. Furthermore, neither method yielded type A crystals alone.

[0064] In methods (a), (b), (c), (e) and (f), an amorphous or syrupy substance was formed immediately after the addition of phosphoric acid, but crystals could be obtained by stirring the amorphous substance at room temperature for 7 to 10 days or by rubbing the syrupy substance frequently for a long time with a spatula. In method (d), a crystal suspension was formed one day after the start of crystallization, but it became a syrupy substance after four days of continued stirring at room temperature. The solvent was evaporated with nitrogen, and the mixture was stirred in ethanol at room temperature overnight, resulting in type B crystals.

[0065] Experimental Example 5 Synthesis of the hydrochloride salt of pyrrolidine compound A [ka] compound 1 (19.43 g) in chloroform (157 mL) 2 (3.3 mL) and acetic acid (2.7 mL) were added and stirred at room temperature for 30 minutes, after which sodium triacetoxyborohydride (10.0 g) was added and stirred at room temperature for 20 hours. An aqueous solution of thorium was added and stirred, and then extracted with chloroform. The resulting organic layer was washed with a saturated aqueous solution of sodium bicarbonate and saturated saline, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by NH silica gel column chromatography (hexane:ethyl acetate=75:25-55:45), followed by silica gel column chromatography (chloroform:methanol=100:0-95:5) and silica gel column chromatography (chloroform:methanol=99:1-96:4) to obtain compound 1. 3 (21.92 g) was obtained as a colorless powder (MS (ESI): m / z 690 [M+H] + ).

[0066] compound 3Aqueous sodium hydroxide solution (2 mol / L, 63.6 mL) was added to a solution of (21.91 g) in methanol (200 mL) and stirred at room temperature for 3 hours. After adding aqueous hydrochloric acid (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 the mixture was stirred and extracted with ethyl acetate. The obtained organic layer was washed with saturated saline, 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), dissolved in ethyl acetate, and phosphate buffer solution (0.1 mol / L, 300 mL) was added and stirred at room temperature for 3 hours. After extraction with ethyl acetate, the organic layer was washed with saturated saline, dried over magnesium sulfate, and concentrated under reduced pressure to obtain compound 1. 4 (17.40 g) was obtained as a colorless powder (MS (ESI): m / z 676 [M+H] + ).

[0067] compound 4 To a solution of (17.40 g) in ethyl acetate (250 mL), hydrochloric acid-ethyl acetate solution (4 mol / L, 31.8 mL) was added and stirred at room temperature for 30 minutes. The solvent was removed by distillation under reduced pressure, diethyl ether was added, and the mixture was stirred, filtered, and dried under reduced pressure to obtain compound 4 The hydrochloride salt (17.43 g) was obtained as a colorless powder (MS (ESI): m / z 676 [M + H] + ).

[0068] Experimental Example 6 Comparison of amorphous pyrrolidine compound A hydrochloride, amorphous pyrrolidine compound A, crystals of pyrrolidine compound A, and crystals of a mixture containing pyrrolidine compound A with reproducibility The hydrochloride of pyrrolidine compound A described in Example 19 of Patent Document 1 is amorphous, and no crystals of the hydrochloride of pyrrolidine compound A were obtained in the subsequent studies (see Experimental Examples 2 and 3 above), so the amorphous hydrochloride of pyrrolidine compound A was used in the comparative test. 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.

[0069] In the comparative test results shown below, the free amorphous form is the amorphous form of pyrrolidine compound A, and the compound obtained by the method described in Example 1. 4 The results were as follows.

[0070] In the comparative test results shown below, the free form crystals are crystals of the free form of pyrrolidine compound A, and it has been confirmed in Experimental Example 1 that the residual solvent in these crystals cannot be reduced to below the standard value set by ICH, and that these crystals are not suitable as pharmaceutical active ingredients.

[0071] In the comparative test results shown below, phosphoric acid type A+B crystals show the results of crystals obtained by method (a) described in Example 3, phosphoric acid type B crystals show the results of crystals of the present invention obtained by the method described in Example 1, phosphoric acid type C crystals show the results of crystals obtained by method (f) described in Example 3, and maleic acid type F crystals show the results of maleic acid type F crystals obtained by the method described in Experimental Example 4.

[0072] In the comparative test results shown below, the results of the malonic acid H-type crystals obtained by the method described in Experimental Example 4 are shown. In addition to the H-type, malonic acid has been able to obtain G-type and I-type crystals, all of which contain toluene in the molecule. Toluene has an effect on the central nervous system. These compounds are known to cause damage to the nervous system, and their inclusion in pharmaceutical ingredients is undesirable from a safety standpoint. Furthermore, it is clear that forms G and I are not suitable as pharmaceutical ingredients, so comparative studies were not conducted on these forms.

[0073] The thermal stability, hygroscopicity, deliquescence, and chemical stability of each of the above crystals were evaluated.

[0074] <Thermal stability evaluation> The evaluation was performed under the following conditions using a thermogravimetry / differential thermal simultaneous measurement device TG / DTA7200 (SII NanoTechnology Inc.). Heating rate: 10K / min Atmosphere: Nitrogen 200mL / min

[0075] <Evaluation of hygroscopicity and deliquescence> The moisture adsorption was evaluated using a moisture adsorption measuring device DVS-1 or DVS-intrinsic (Surface Measurement Systems Limited) as follows. The tare weight of the sample was corrected in advance. The weight was then precisely measured at the start of the measurement by placing the sample in a cell and hanging it on the precision balance of the device. The weight change was recorded over time as the humidity was gradually changed, and the equilibrium weight at each humidity was calculated. The rate of weight change at each humidity was calculated based on the anhydrous weight calculated from the moisture content at the start of the measurement, either when dry (0% RH) or as determined by a different method.

[0076] <Chemical stability evaluation> The samples were stored at 60°C in a sealed state and at 60°C, 75% RH for one week. The increase or decrease in the amount of related substances before and after storage was calculated from the area percentage of each peak by high performance liquid chromatography, and the condition after storage was also observed. The results are shown in Table 7 below.

[0077] [Table 7]

[0078] As reproducible solids of pyrrolidine compound A and mixtures containing it, hydrochloride amorphous, free form amorphous, free form crystalline, phosphoric acid type A+B crystalline, phosphoric acid type B, which is one embodiment of the crystal of the present invention, Acid B type crystals, phosphoric acid C type crystals, maleic acid F type crystals, malonic acid G type crystals, malonic acid H type crystals, and malonic acid I type crystals have been found. Among these, the free form crystals could not reduce the residual solvent to below the standard value set by ICH, and malonic acid G type crystals and malonic acid I type crystals contain toluene in the molecule, and are not suitable crystals as pharmaceutical raw materials from the viewpoint of safety. Similarly, maleic acid F type crystals could not completely remove the residual solvent toluene. On the other hand, no residual solvent was confirmed in the crystals of the present invention, and since they do not contain toluene in the molecule, they are considered to be crystals that are not problematic from the viewpoint of safety.

[0079] In addition, the amorphous hydrochloride, phosphate C type crystals, maleic acid F type crystals, and malonic acid H type crystals were all found to be deliquescent, but the phosphate B type crystals did not show any deliquescent properties, and the weight change at 90% RH was less than 1%, and even after storage at 60°C and 75% RH for one week, the increase in related substances was 0.05%, making them stable to humidity and very chemically stable crystals.

[0080] Furthermore, both the free amorphous form and the phosphoric acid A+B crystals showed a weight change of 2% or more when heated to 130°C, but the phosphoric acid B crystals showed no weight change even at temperatures above 200°C, indicating that they were crystals with excellent thermal stability.

[0081] Experimental Example 7: Study of precipitation conditions for the crystals of the present invention [ka]

[0082] Condition 1 compound 5 To a solution of (15.00 g) in ethyl acetate (67.75 g), an aqueous potassium carbonate solution (potassium carbonate 2.64 g, purified water 75 mL) was added dropwise at 25°C and the mixture was stirred at 300 rpm for 30 minutes. The mixture was stirred at RT. 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 reached 27 mL. This process was repeated twice. Ethanol (24.32 g) was added to the resulting concentrated mixture until the volume reached 45 mL. 24% sodium hydroxide aqueous solution (4.79 g) and purified water (12 mL) were added dropwise at 25°C. The reaction mixture was stirred at 300 rpm for 6 hours. An 85% aqueous solution of phosphoric acid (6.61 g) and purified water (22.5 mL, 4 (1.5 times the volume of the weight of the sample) was dripped over 5 minutes. Then, purified water (90 mL, 4 (6.0 times the weight of the product by volume) for 30 minutes. After the dropwise addition of purified water was completed, the mixture was stirred for 30 minutes, the reaction temperature was increased to 35°C, and seed crystals (0.3746 g, compound 4 After 10 hours from the addition of the seed crystals, the reaction temperature was increased to 20° C. and the mixture was stirred for another 30 minutes. The insoluble matter was filtered off and dried under reduced pressure to obtain the present invention crystals (7.48 g). The obtained crystals had poor filterability, and filtration took a long time. In addition, the wet material before drying became a slurry, which made it difficult to work with. The particle size of the crystals obtained was measured, and the mode diameter was found to be about 6 μm. The particle size distribution of the crystals obtained under condition 1 was measured (Figure 5-1).

[0083] Condition 2 compound 3 To a solution (53.20 g) of (17.58 g) in ethanol, an aqueous solution of sodium hydroxide (24%, 4.67 g) and purified water (16.20 g) were added and stirred at 40°C for 3 hours to obtain compound (17.58 g). 4 The resulting solution (73.14 g) was 4 To 18.28 g of the solution, add phosphoric acid (1.91 g, 2.6 equivalents) and purified water (7.49 g, compound 4 For weight After stirring for 1 hour, seed crystals (125 mg, compound 4 After 17 hours and 30 minutes from the addition of the seed crystals, purified water (30.0 mL, compound 4 (6.0 times the volume of the product) After 10 hours, the reaction temperature was increased to 20° C. and stirring was continued. After 27 hours, the solid was collected by filtration and dissolved in purified water (25.23 g, 4 The solid was washed with water (5 times the volume of the solid relative to the weight of the solid) at 50° C. to obtain the present invention crystal (4.01 g). Sampling was performed 16 and 21 hours after the addition of the seed crystals, and the crystal precipitation rates were calculated by HPLC and were found to be 4% and 97%, respectively. It was found that when the amount of purified water before seed inoculation was small as in condition 2, the crystal precipitation rate was extremely low. That is, it was found that the amount of purified water added to the ethanol solution, i.e., the composition of the crystallization solvent, is important for precipitating the crystals of the present invention. In addition, when purified water was further added to a solution with a low precipitation rate, crystals were precipitated, but the obtained crystals had poor operability such as filtration, as in condition 1.

[0084] Condition 3 compound 3 To a solution (79.82 g) of (26.37 g) in ethanol, add an aqueous solution of sodium hydroxide (24%, 7.01 g) and purified water (24.30 g) and stir at 40°C for 4 hours and 30 minutes to obtain compound (III). 4 The solution of the compound obtained was 4 To a solution of phosphoric acid (9.70 g, 2.2 equivalents) and purified water (45.00 g, compound 4 Add purified water (45.13 g, 1.5 times the volume of the compound) and 4 (1.5 times the volume of the compound) was added and stirred at 30°C for 40 minutes, and then seed crystals (751 mg, compound 4 After 16 hours, purified water (135 mL, compound A) was added over a period of 2 hours. 4 After 6 hours, the temperature was increased to 20° C., and the mixture was stirred for another hour. The solid was collected by filtration, and then purified water (150.02 g, 4 The solid was dried at 50° C. to obtain the present invention crystals (26.95 g). The particle size of the obtained crystals was measured, and the mode diameter was about 10 μm. The obtained crystals had good operability for filtration, etc. The particle size distribution of the crystals obtained under condition 3 was measured (Figure 5-2). As described above, the temperature was set to about 30°C, and the amount of purified water described in condition 3 was added before the seed crystals were added. After allowing a certain amount of time for the crystals to grow large, further purified water was added, thereby obtaining crystals with good precipitation rate and ease of operation.

[0085] [Table 8]

[0086] Reactor and stirring conditions Condition 1 Reactor: EasyMax (registered trademark) (Mettler Toledo) Stirring conditions: 300 rpm

[0087] Condition 2 Reactor: OptyMax (registered trademark) (Mettler Toledo) Stirring conditions: 250 rpm

[0088] HPLC measurement conditions Equipment name: Column: GL Science, Inertsil ODS-3V (5 μm, 4.6 x 150 mm) Fluidized bed A: Water / acetonitrile / trifluoroacetic acid = 1900:100:1 Fluidized bed B: Water / acetonitrile / trifluoroacetic acid = 100:1900:1

[0089] Particle trend measurement conditions Equipment name: Particle Track (registered trademark) MALVERN, Mastersizer 2000 (wet type) Measurement range: 0.020~2000.000μm Measurement time: 10 seconds Measurement intensity range: 3.0~20.0% Stirring speed: approx. 2000 rpm

[0090] As mentioned above, by adding sufficient purified water before adding seed crystals as in condition 2, and then allowing the crystals to grow for a certain period of time before adding more purified water, it was possible to obtain crystals containing many large-cord crystals with good fluidity and filterability while ensuring a sufficient amount of precipitation. This improved operability such as filtration.

[0091] Experimental Example 8 Human MC1R agonist measurement Using the crystals of the present invention, the intracellular cAMP concentration was measured according to the following method described in Experimental Example 1 of Patent Document 1, and EC 50 The value was calculated. (1) Cell culture method The human melanoma cell line HBL was used to measure human MC1R agonist activity. HBL was cultured in F-10 containing 10% FCS and penicillin-streptomycin. Culture was performed using Nutrient Mixture. (2) cAMP assay and data calculation Compound solutions of each concentration were mixed with cAMP assay buffer (10 mM HEPES, 0.1% BSA-containing HBSS (Hank's Balanced Salt Solution)) and plated in a 96-well plate. HBL was 5 × 10 4 The cells were suspended in cAMP assay buffer containing 0.5 mM IBMX to a concentration of 1 / mL, dispensed into the 96-well plate, mixed, and left to stand at 37°C for 30 minutes, after which the intracellular cAMP concentration was measured by a fluorescence method using Envision (ex. 320 nm, em. 590 nm and 665 nm). The quantitative value of the cAMP concentration was calculated using Prism 5.02 from the obtained data ratio value (665 nm measurement value / 590 nm measurement value x 10000), and the induction% value (the average cAMP concentration of the vehicle was 0%, and that of αMSH was 100%) was used to calculate the cAMP concentration. -6 The EC% of each sample was calculated based on the average cAMP concentration at M taken as 100%. 50 values ​​were calculated. As a result, the crystal of the present invention was EC 50 The agonistic activity of the crystals was 5.3 nM, indicating that the crystals had strong human MC1R agonist activity. [Industrial Applicability]

[0092] The crystal of the present invention is free of residual solvent used in obtaining the crystal, has excellent thermal stability, is stable with little weight change in relation to humidity, does not deliquesce, has excellent chemical stability, and from the viewpoint of safety, is a crystal that does not contain compounds that may have adverse effects on living organisms. Furthermore, the crystal can be obtained by an industrially suitable method with good reproducibility, and therefore is an excellent crystal to be used as a pharmaceutical active ingredient.

Claims

[Claim 1] The invention described herein

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