Crystalline form of cyclohexenone compounds

Abstract

The purpose of the present disclosure is to provide a crystal form of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one, this crystal form being of superior stability and desirable in terms of production. The present disclosure provides a crystal form which, in the powder X-ray diffraction spectrum (CuKα), demonstrates diffraction angles (2θ ± 0.2°) that have characteristic peaks at 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8°.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims priority based on Japanese Patent Application No. 2022 - 019058 filed on February 9, 2022 (the entire disclosure of which is incorporated herein by reference). The present disclosure relates to a crystalline form of a cyclohexenone compound useful as a pharmaceutical active ingredient and a pharmaceutical composition containing the crystalline form.

Background Art

[0002] Neurodegenerative diseases are roughly classified into central and peripheral types depending on the affected nerve site. Representative central nervous system diseases include Alzheimer's disease, amyotrophic lateral sclerosis, and spinal cord injury. On the other hand, representative diseases in which peripheral nerves are damaged include sensory nerve disorders such as neuropathic pain and sensory dulling, and autonomic nerve disorders accompanied by constipation, urinary disorders, etc. Although the pathogenesis of the pathological conditions in neurodegenerative diseases is various, it is considered that degeneration and atrophy of neurites and / or neuronal cell death occur. At present, there are few fundamental therapeutic drugs for improving neuropathy for these diseases.

[0003] Patent Document 1 discloses that 3-(15 - hydroxypentadecyl)-2,4,4 - trimethylcyclohex - 2 - en - 1 - one exhibits a nerve growth promoting effect and is useful as a pharmaceutical for preventing and treating brain diseases such as dementia. Patent Document 2 discloses that the same compound is useful as a therapeutic agent for urinary disorders. However, 3-(15 - hydroxypentadecyl)-2,4,4 - trimethylcyclohex - 2 - en - 1 - one has insufficient pharmacokinetics in normal formulations (Patent Document 3).

[0004] Also, generally, when a compound is used as an active ingredient or a pharmaceutical active ingredient of a pharmaceutical product, chemical and physical stability of the compound is required to maintain quality stably and / or to facilitate storage management.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] International Publication No. WO99 / 08987 [Patent Document 2] International Publication No. WO2002 / 066024 [Patent Document 3] International Publication No. WO2013 / 147072 [Overview of the project] [Problems that the invention aims to solve]

[0006] This disclosure aims to provide a novel compound having excellent neurite growth activity. Furthermore, this disclosure aims to provide a stable crystalline form of the novel compound that is useful as an active ingredient or drug substance for pharmaceuticals. [Means for solving the problem]

[0007] In order to solve the aforementioned problem, after diligent consideration, the following formula (I) was obtained.

[0008] [ka]

[0009] A novel cyclohexenone compound represented by (chemical name: 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one, hereinafter also referred to as "compound (I)") has been found to have nerve growth-promoting effects (neurite growth and / or effects that increase the proportion of neurite cells) and is useful as a medicine for treating diseases that improve by promoting nerve growth, including neurodegenerative diseases, pain, and / or lower urinary tract disorders. Furthermore, compound (I) has been found to exist in three crystalline forms (Type I crystal, Type II crystal, and Type III crystal), and these crystalline forms have low hygroscopicity and excellent solid stability.

[0010] In other words, this disclosure provides the following [1] to

[15] .

[0011] [1] A crystalline form of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one having at least three peaks with diffraction angles (2θ±0.2°) selected from 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8° in its powder X-ray diffraction spectrum (CuKα).

[0012] [2] The crystal form described in [1], wherein the powder X-ray diffraction spectrum (CuKα) has peaks at diffraction angles (2θ±0.2°) of 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8°.

[0013] [3] The crystal form according to [1] or [2], wherein the powder X-ray diffraction spectrum is substantially the same as that shown in Figure 1.

[0014] [4] A crystal form according to any of [1] to [3], having an endothermic peak with a peak temperature of around 35°C in differential scanning calorimetry (DSC measurement).

[0015] A pharmaceutical composition containing the crystalline form described in any of [5], [1], to [4].

[0016] [6] A crystalline form of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one having at least five peaks with diffraction angles (2θ±0.2°) selected from 6.1°, 11.4°, 12.5°, 15.1°, 19.4°, and 24.0° in powder X-ray diffraction spectrum (CuKα).

[0017] [7] The crystal form described in [6], wherein the powder X-ray diffraction spectrum (CuKα) has peaks at diffraction angles (2θ±0.2°) of 6.1°, 11.4°, 12.5°, 15.1°, 19.4°, and 24.0°.

[0018] [8] The crystal form according to [6] or [7], wherein the powder X-ray diffraction spectrum is substantially the same as that shown in Figure 3.

[0019] 〔9〕A crystalline form as described in any one of 〔6〕~〔8〕, having an endothermic peak with a peak temperature (peak top value) around 34 °C in differential scanning calorimetry (DSC measurement).

[0020] 〔10〕A pharmaceutical composition containing the crystalline form as described in any one of 〔6〕~〔9〕.

[0021] 〔11〕A crystalline form of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohex-2-en-1-one having at least 5 or more peaks selected from diffraction angles (2θ±0.2°) of 11.2°, 12.4°, 14.1°, 14.9°, 18.8° and 19.4° in a powder X-ray diffraction spectrum (CuKα).

[0022] 〔12〕The crystalline form as described in 〔11〕, having peaks at diffraction angles (2θ±0.2°) of 11.2°, 12.4°, 14.1°, 14.9°, 18.8° and 19.4° in a powder X-ray diffraction spectrum (CuKα).

[0023] 〔13〕The crystalline form as described in 〔11〕 or 〔12〕, wherein the powder X-ray diffraction spectrum is substantially the same as that shown in FIG. 5.

[0024] 〔14〕A crystalline form as described in any one of 〔11〕~〔13〕, having an endothermic peak with a peak temperature (peak top value) around 34 °C in differential scanning calorimetry (DSC measurement).

[0025] 〔15〕A pharmaceutical composition containing the crystalline form as described in any one of 〔11〕~〔14〕.

Advantages of the Invention

[0026] This disclosure provides compound (I), which is useful as a compound having nerve growth promoting activity. It also provides compound (I), which is useful as a compound having excellent pharmacokinetic and / or urinary function improving activity. Furthermore, compound (I) in this disclosure is a novel compound, and its crystal form, stable crystal form of compound (I), and method of producing it are currently unknown. In this disclosure, the crystal form of compound (I) (Type I crystal, Type II crystal, Type III crystal) is a useful form when using the compound as a pharmaceutical active pharmaceutical ingredient because it is superior to other crystal forms in terms of, for example, handling (lower hygroscopicity) and / or quality control.

[0027] Furthermore, the Type I, Type II, and Type III crystals of this disclosure have residual solvent levels below the standard values ​​set forth in the ICH guidelines for residual solvents in pharmaceuticals, and are therefore safe as pharmaceuticals. [Brief explanation of the drawing]

[0028] [Figure 1] The powder X-ray diffraction spectrum (CuKα) of the type I crystal of compound (I) obtained in Example 1 is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)). [Figure 2] The differential scanning calorimetry (DSC) curve of the type I crystal of compound (I) obtained in Example 1 is shown. [Figure 3] The powder X-ray diffraction spectrum (CuKα) of the type II crystal of compound (I) obtained in Example 2 is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)). [Figure 4] The differential scanning calorimetry (DSC) curve of the type II crystal of compound (I) obtained in Example 2 is shown. [Figure 5] The powder X-ray diffraction spectrum (CuKα) of the type III crystal of compound (I) obtained in Example 3 is shown (the vertical axis represents intensity (counts), and the horizontal axis represents diffraction angle (2θ)). [Figure 6] The differential scanning calorimetry (DSC) curve of the type III crystal of compound (I) obtained in Example 3 is shown. [Figure 7]The water adsorption / desorption isotherm of the type I crystal of compound (I) obtained in Example 1 is shown. [Figure 8] The water adsorption / desorption isotherm of the type II crystal of compound (I) obtained in Example 2 is shown. [Figure 9] The water adsorption / desorption isotherm of the type III crystal of compound (I) obtained in Example 3 is shown. [Figure 10] Graphs showing residual urine volume in each group. Values ​​represent the mean ± SE. The n size for each group is 10, 10, 12, and 12 from left to right. *: p<0.05, Dunnett's test shows a significant difference compared to Vehicle. NS: Dunnett's test shows no significant difference compared to Vehicle. #: p<0.05, Student's t-test shows a significant difference compared to compound (I).##: p<0.01, Student's t-test shows a significant difference compared to Vehicle. [Modes for carrying out the invention]

[0029] In this specification, the singular form (a, an, the, etc.) includes both singular and plural forms unless otherwise explicitly stated herein or the context clearly contradicts it. Unless otherwise explicitly stated otherwise, when "compound (I)" is mentioned herein, it means 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one and is used to mean both "amorphous" and "crystalline" forms.

[0030] Compound (I) can be synthesized, for example, by the method described in Synthesis Example 1 below, but is not limited to this method.

[0031] In this specification, the terms “crystal” and “amorphous” are used in their ordinary sense within the art of the pharmaceutical field to which this disclosure belongs. In this specification, the term “crystal” and related terms used herein, such as “crystalline form” and “crystalline morphology,” are used interchangeably. Similarly, in this specification, the term “amorphous” and related terms used herein, such as “amorphous body” and “amorphous morphology,” are used interchangeably.

[0032] The crystals of this disclosure can be isolated and purified by well-known separation and purification techniques such as recrystallization, crystallization, distillation, and column chromatography.

[0033] Where used herein, and unless otherwise specified, the term “crystal” and related terms herein, when used to describe a compound, substance, modification, material, component, or product, mean that the compound, substance, modification, material, component, or product is substantially crystalline as determined by X-ray diffraction. See, for example, Remington: The Science and Practice of Pharmacy, 21st edition; Lippincott, Williams and Wilkins, Baltimore, MD (2005); United States Pharmacopeia, 23rd edition, 1843–1844 (1995).

[0034] As used herein, and unless otherwise specified, the term “crystalline form” and related terms herein mean a crystalline solid form. Crystalline forms include single-component and multi-component crystalline forms and may optionally, but not limited to, cocrystals, salts (including pharmaceutically acceptable salts), polymorphs, solvates, hydrates, and / or other molecular complexes. In certain embodiments, the crystalline form of a substance may substantially not include amorphous and / or other crystalline forms. In certain embodiments, the crystalline form of a substance may contain one or more amorphous and / or other crystalline forms by weight of less than 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50%.

[0035] The term "cocrystal" refers to a molecular complex derived from numerous cocrystal-forming materials known in the art. Unlike salts, cocrystals typically do not involve hydrogen transfers between the cocrystal-forming material and the drug; instead, they involve intermolecular interactions such as hydrogen bonds, aromatic ring stacks, or dispersion forces between the cocrystal-forming material and compound (I) in the crystalline structure.

[0036] Where used herein, and unless otherwise specified, the terms “polymorph,” “polymorphic form,” and related terms herein mean two or more crystalline forms consisting of substantially the same molecule, group of molecules, and / or ions. Different crystalline forms, as well as different polymorphs, may have different properties, such as melting point, heat of fusion, solubility, solubility characteristics, and / or vibrational spectrum (as a result of the arrangement or conformation of molecules and / or ions in the crystal lattice). Differences in properties may affect pharmaceutical parameters such as storage stability, compressibility and density (important in formulation and product manufacturing), and dissolution rate (an important factor in bioavailability). Differences in stability can arise from changes in chemical reactivity (e.g., differential oxidation, where the dosage form discolors faster when composed of one polymorph than when composed of another), mechanical changes (e.g., a tablet disintegrating during storage if a kinetically preferred polymorph transforms into a thermodynamically more stable polymorph), or both (e.g., tablets of one polymorph are more prone to decomposition at high humidity). As a result of differences in solubility, in extreme cases, some solid transitions may lead to a loss of potency, and in other extreme cases, toxicity may result. In addition, physical properties can be important in processing (e.g., some polymorphs may be relatively prone to forming solvates, or may be difficult to filter and wash to remove impurities, and particle shape and size distribution may differ between polymorphs).

[0037] As used herein, and unless otherwise specified, the terms “amorphous,” “amorphous form,” and related terms used herein mean that the substance, component, or product is substantially non-crystalline as determined by X-ray diffraction. In particular, the term “amorphous form” describes an irregular solid form, i.e., a solid form lacking long-range crystalline order.

[0038] The term "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions known in the art. Pharmaceutically acceptable salts may be safe for ingestion by animals or humans.

[0039] Techniques for characterizing crystalline and amorphous morphologies include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single-crystal X-ray diffraction, vibrational spectroscopy, such as infrared (IR) and Raman spectroscopy, solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot-stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility measurement, dissolution measurement, atomic analysis, and Karl Fischer analysis. Characteristic unit cell parameters can be determined using one or more techniques, including, but are not limited to, single-crystal diffraction and powder diffraction, as well as X-ray diffraction and neutron diffraction. Useful techniques for analyzing powder diffraction data include profile refinement, such as Rietveld refinement, which can be used, for example, to analyze diffraction peaks associated with a single phase in a sample containing two or more solid phases. Another useful method for analyzing powder diffraction data is unit cell indexing, which allows those skilled in the art to determine unit cell parameters from a sample containing crystalline powder.

[0040] Furthermore, due to the nature of the data, the diffraction angle and overall pattern of powder X-ray diffraction patterns may be important when determining the identity of a crystal. The relative intensity of the powder X-ray diffraction pattern can vary somewhat depending on the crystal growth direction, particle size, measurement conditions, etc., and therefore should not be interpreted strictly.

[0041] The numerical values ​​obtained from various patterns may have some error depending on the direction of crystal growth, particle size, measurement conditions, etc. Therefore, the numerical values ​​of the diffraction angle (2θ) in the powder X-ray diffraction patterns in this specification may have a measurement error of approximately ±0.2°.

[0042] Furthermore, the endothermic peak in the differential scanning calorimetry (DSC measurement) curve may vary depending on the measurement temperature, such as the rate of temperature increase per minute and the purity of the sample. In this specification, the term "around" in relation to the DSC measurement temperature value means a temperature value within ±2, 3, 4, or 5°C of that value.

[0043] In one embodiment of the present disclosure, a type I crystal of compound (I) has, for example, the powder X-ray diffraction spectrum (CuKα) shown in Figure 1, and the differential scanning calorimetry (DSC measurement) curve shown in Figure 2.

[0044] Here, characteristic peaks in the powder X-ray diffraction spectrum (CuKα) of the type I crystal of compound (I) can be found at diffraction angles (2θ±0.2°) of 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8°.

[0045] The type I crystal of compound (I) according to this disclosure is a crystal form having at least three or more peaks selected from the above peaks, preferably a crystal form having at least five or more peaks selected from the above peaks, and particularly preferably a crystal form having any of the above peaks.

[0046] Furthermore, examples of endothermic peaks in the differential scanning calorimetry (DSC measurement) curve of the type I crystal of compound (I) include those around 33°C to 37°C, preferably around 34°C to 36°C, and more preferably around 35°C.

[0047] The crystalline morphology relating to this disclosure may be characterized by a combination of the embodiments described above, and these combinations are not inconsistent. For example, the crystalline morphology may be characterized by a combination of the above-described peaks in the X-ray diffraction pattern and the above-described endothermic peaks measured by DSC.

[0048] In some embodiments, the crystalline morphology disclosed herein may comprise a plurality of crystals (polymorphs) having a spatially regular atomic arrangement, resulting in different physicochemical properties. In some embodiments, the crystalline morphology disclosed herein may be a mixture containing polymorphs.

[0049] In some embodiments, the crystalline form of compound (I) disclosed herein may be physically and / or chemically pure. In certain embodiments, the crystalline form of the compound disclosed herein may be at least about 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, or 80% physically and / or chemically pure. In some embodiments, the crystalline form of the compound disclosed herein is substantially pure. As used herein, and unless otherwise specified, a sample containing a particular crystalline or amorphous form that is "substantially pure," for example, substantially free of other solid forms and / or other compounds, contains, in certain embodiments, one or more other solid forms and / or other compounds in amounts of about 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25%, or less than 0.1% by weight.

[0050] In some embodiments, the crystalline form of compound (I) disclosed herein is stable upon exposure to conditions of approximately 1, 2, 3, 4, 5, 6, 7, 8, 12, 16, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 144, or 156 weeks or longer and approximately 160, 156, 150, 138, 126, 114, 102, 90, 78, 66, 54, 42, 30, 24, 18, 12, or 6 weeks or less at approximately 25, 30, 35°C and approximately 65, 70, 75, 80, or 85% relative humidity. These conditions may be closed or open conditions. As used herein, “closed” conditions may mean closing or sealing the lid of the bottle containing the sample during the stability experiment, and “open” conditions may mean the lid is open. In some embodiments, the crystalline form of compound (I) disclosed herein is stable when exposed to conditions of approximately 25°C and approximately 60% relative humidity for approximately 4 weeks. In some embodiments, these conditions are closed conditions. In other words, the crystalline form of the compounds disclosed herein exhibits excellent storage stability over long periods. In this specification, “stable” means that the increase in impurities compared to the initial amount of impurities is approximately 1.0, 0.5, 0.3, 0.1, 0.05, or 0.01% or less. Stability can also be demonstrated, for example, by the X-ray diffraction pattern maintaining 10, 20, 30, 40, 50, 60, 70, 80, or 90% or more of the initial peak at (2θ±0.2°).

[0051] Furthermore, the crystalline form of the disclosed material may be a solvate (e.g., a hydrate) or a non-solvate. Compounds labeled with isotopes (e.g., deuterium, 3H, 14C, 35S, 125I, etc.) are also included in the compounds described herein or their pharmaceutically acceptable salts.

[0052] In one embodiment of the present disclosure, type I crystals of compound (I) are obtained, for example, by step (i-1) adding compound (I) to solvent I-1 and dissolving it, and Step (i-2): Add solvent I-2 to the solution of compound (I) obtained in step (i-1), stir, and crystallize compound (I). It can be obtained by a method including [a certain component]. In such embodiments, examples of solvent I-1 include methanol, ethanol, 1-propanol, 2-propanol, acetone, and the like.

[0053] In step (i-1), the amount of compound (I) added is not particularly limited, but for example, 0.1 to 20 g, preferably 0.5 to 5 g, of compound (I) by mass can be added to 10 mL of solvent I-1. The temperature in step (i-1) is set appropriately depending on the solvent used, and is set between 20°C and the boiling point of the solvent.

[0054] In this embodiment, the solvent I-2 can be, for example, water. The ratio (v / v) of solvent I-1 to solvent I-2 can be, for example, 0 to 50 ml, preferably 0 to 20 ml, of solvent I-2 to 10 ml of solvent I-1. The temperature in the crystallization of step (i-2) is set appropriately depending on the solvent used, and is set between -20°C and 30°C.

[0055] Alternatively, solvents I-1 and I-2 can be added simultaneously, heated to dissolve, and then cooled and stirred to crystallize compound (I).

[0056] Stirring in the crystallization process (i-2) is carried out using a stirrer, stirring blades, magnetic stirrer, etc., as appropriate depending on the amount of solvent, the size of the reaction vessel, etc. The stirring speed is usually 1 to 1200 rpm, and preferably 10 to 600 rpm.

[0057] Furthermore, the type I crystals obtained as described above have a chemical purity of 90% or more of compound (I) and can be measured by high-performance liquid chromatography (HPLC). Preferably, the type I crystals have a chemical purity of 95% or more of compound (I), and more preferably, 99% or more are type I crystals.

[0058] The type I crystals of compound (I) may be any type I crystals, and may be single type I crystals or polymorphic mixtures containing other types of crystals. Preferably, 90% by weight or more are type I crystals, and more preferably 95% or more are type I crystals.

[0059] In one embodiment of the present disclosure, a type II crystal of compound (I) has, for example, the powder X-ray diffraction spectrum (CuKα) shown in Figure 3, and the differential scanning calorimetry (DSC measurement) curve shown in Figure 4.

[0060] Here, characteristic peaks in the powder X-ray diffraction spectrum (CuKα) of type II crystals of compound (I) can be found at diffraction angles (2θ±0.2°) of 6.1°, 11.4°, 12.5°, 15.1°, 19.4°, and 24.0°.

[0061] The type II crystal of compound (I) according to this disclosure is a crystal having at least five peaks selected from the above peaks, and is particularly preferably a crystal having any of the above peaks.

[0062] Furthermore, the endothermic peak in the differential scanning calorimetry (DSC measurement) curve of the type II crystal of compound (I) can be found, for example, around 32°C to 36°C, preferably around 33°C to 35°C, and more preferably around 34°C.

[0063] In one embodiment of the present disclosure, a type II crystal of compound (I) is obtained, for example, by step (ii-1) adding compound (I) to solvent II and dissolving it, and Step (ii-2): A step to cool the solution of compound (I) obtained in step (ii-1) and crystallize compound (I). It can be obtained by a method including [a certain component]. In such embodiments, solvent II may be, for example, acetonitrile.

[0064] In step (ii-1), the amount of compound (I) added is not particularly limited, but for example, 1 to 4 g, preferably 1.5 to 3.5 g, of compound (I) can be added by mass to 10 mL of solvent II. The temperature in step (ii-1) is set as appropriate, between 15 and 40°C. The temperature in crystallization in step (ii-2) is set between -10 and 5°C. Preferably, the temperature is cooled from the temperature in step (ii-1) to less than 3°C and maintained at a temperature below 3°C.

[0065] Crystallization in step (ii-2) may be carried out under stirring. In this case, stirring should be performed using a stirrer, stirring blades, etc., as appropriate depending on the amount of solvent, the size of the reaction vessel, etc. Stirring can also be performed, for example, by shaking the reaction vessel. The stirring speed is usually 1 to 600 rpm, and 10 to 100 rpm is preferred.

[0066] Furthermore, the type II crystals obtained as described above have a chemical purity of 90% or more of compound (I) and can be measured by high-performance liquid chromatography (HPLC). Preferably, the type II crystals have a chemical purity of 95% or more of compound (I), and more preferably, 99% or more are type II crystals.

[0067] The type II crystals of compound (I) may be any type containing type II crystals, and may be single type II crystals or polymorphic mixtures containing other types of crystals. Preferably, 90% by weight or more are type II crystals, and more preferably 95% or more are type II crystals.

[0068] In one embodiment of the present disclosure, a type III crystal of compound (I) has, for example, the powder X-ray diffraction spectrum (CuKα) shown in Figure 5, and the differential scanning calorimetry (DSC measurement) curve shown in Figure 6.

[0069] Here, characteristic peaks in the powder X-ray diffraction spectrum (CuKα) of compound (I) type III crystals can be found at diffraction angles (2θ±0.2°) of 11.2°, 12.4°, 14.1°, 14.9°, 18.8°, and 19.4°.

[0070] The type III crystal of compound (I) according to this disclosure is a crystal having at least five peaks selected from the above peaks, and is particularly preferably a crystal having any of the above peaks.

[0071] Furthermore, the endothermic peak in the differential scanning calorimetry (DSC measurement) curve of the type III crystal of compound (I) can be found, for example, around 32°C to 36°C, preferably around 33°C to 35°C, and more preferably around 34°C.

[0072] In one embodiment of the present disclosure, the type III crystal of compound (I) is obtained, for example, by step (iii-1) adding the type II crystal of compound (I) to solvent III-1 and stirring. Step (iii-2) A step of recovering the solid of compound (I) from the suspension of compound (I) obtained in step (iii-1) above. It can be obtained by a method including the above. In such embodiments, solvent III-1 is a mixed solvent of methanol and water. When a mixed solvent of methanol and water is used, the ratio of use (v / v) can be, for example, 1 to 15 ml, preferably 5 to 11 ml, of water per 100 ml of methanol.

[0073] In step (iii-1), the amount of type II crystals of compound (I) added is not particularly limited, but for example, 1 to 5 g, preferably 1.5 to 2.5 g, of compound (I) can be added by mass to 10 mL of solvent III-1. The temperature in step (iii-1) is set appropriately depending on the solvent used, and is set between 0 and 8°C.

[0074] The recovery method in step (iii-2) can be a wide range of methods that can be used in the art to which this disclosure belongs, and includes filtration, washing with an organic solvent, and vacuum drying.

[0075] The stirring in step (iii-1) is carried out using a stirrer, stirring blades, magnetic stirrer, etc., as appropriate depending on the amount of solvent, the size of the reaction vessel, etc. The stirring speed is usually 1 to 600 rpm, and 10 to 300 rpm is preferred.

[0076] Furthermore, the type III crystals obtained as described above have a chemical purity of compound (I) of 90% or more and can be measured by high-performance liquid chromatography (HPLC). Preferably, the type III crystals have a chemical purity of 95% or more of compound (I), and more preferably, 99% or more are type III crystals.

[0077] The type III crystals of compound (I) may be any type that contains type III crystals, and may be single type III crystals or polymorphic mixtures containing other types of crystals. Preferably, 90% by weight or more are type III crystals, and more preferably 95% or more are type III crystals.

[0078] As shown in the examples below, the type I, type II, and type III crystals of compound (I) relating to this disclosure all have low hygroscopicity and excellent solid stability (storage stability, etc.). Solid stability of the active pharmaceutical ingredient and the active ingredient in pharmaceuticals is important for both industrial operations and maintaining quality.

[0079] Furthermore, solvents used in the manufacture of pharmaceuticals can be toxic, and from a safety standpoint, it is desirable to minimize the amount of solvent remaining in the manufacturing process. Compound (I)'s type I, type II, and type III crystals do not contain residual solvents exceeding the regulatory limits set by the ICH (International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use) guidelines.

[0080] Therefore, the type I, type II, and type III crystals of compound (I) relating to this disclosure possess excellent properties required for use as a pharmaceutical or pharmaceutical active pharmaceutical ingredient.

[0081] The type I, type II, and type III crystals of compound (I) relating to this disclosure have excellent neurite growth effects and are useful as pharmaceuticals for treating diseases that improve by promoting nerve growth, including neurodegenerative diseases, pain, and / or lower urinary tract disorders, through effects such as promoting neurite growth and / or increasing the proportion of neurite-forming cells.

[0082] When compound (I) is used as a pharmaceutical, type I, type II, and / or type III crystals can be administered in various forms depending on the preventive or therapeutic purpose, with or without grinding the type I, type II, and / or type III crystals. These forms may include oral preparations such as tablets, capsules, granules, fine granules, powders, and dry syrups, as well as parenteral preparations such as suppositories, inhalants, nasal drops, ointments, patches, and injections, with oral preparations being preferred. These pharmaceutical compositions can be manufactured using pharmaceutically acceptable carriers by pharmaceutical methods known and practiced to those skilled in the art.

[0083] As pharmaceutical carriers, various organic or inorganic carrier substances commonly used as formulation materials are used, and are incorporated as excipients, binders, disintegrants, lubricants, and coatings in solid formulations, and as solvents, solubilizers, suspending agents, isotonic agents, buffers, and analgesics in liquid formulations. Furthermore, formulation additives such as preservatives, antioxidants, colorants, sweeteners, and stabilizers may be used as needed.

[0084] Excipients include lactose, sucrose, D-mannitol, starch, crystalline cellulose, and calcium silicate.

[0085] Examples of binders include hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone, syrup powder, and hypromellose.

[0086] Examples of disintegrants include sodium starch glycolate, carmellose calcium, croscarmellose sodium, crospovidone, low-substituted hydroxypropyl cellulose, and partially pregelatinized starch.

[0087] Examples of lubricants include talc, magnesium stearate, sucrose fatty acid esters, stearic acid, and sodium stearyl fumarate.

[0088] Examples of coating agents include ethylcellulose, aminoalkyl methacrylate copolymer RS, hypromellose, and sucrose.

[0089] Examples of solvents include water, propylene glycol, and physiological saline.

[0090] Examples of solubilizers include polyethylene glycol, ethanol, α-cyclodextrin, macrogol 400, and polysorbate 80.

[0091] Examples of suspending agents include carrageenan, crystalline cellulose / carmellose sodium, and polyoxyethylene hydrogenated castor oil.

[0092] Examples of isotonic agents include sodium chloride, glycerin, and potassium chloride.

[0093] Examples of pH adjusters and buffering agents include sodium citrate, hydrochloric acid, lactic acid, phosphoric acid, and sodium dihydrogen phosphate.

[0094] Examples of pain relievers include procaine hydrochloride and lidocaine.

[0095] Examples of preservatives include ethyl parahydroxybenzoate, cresol, and benzalkonium chloride.

[0096] Examples of antioxidants include sodium sulfite, ascorbic acid, and natural vitamin E.

[0097] Examples of coloring agents include titanium dioxide, ferric oxide, food coloring blue No. 1, and copper chlorophyll.

[0098] Examples of flavoring and odor-masking agents include aspartame, saccharin, sucralose, l-menthol, and mint flavor.

[0099] Examples of stabilizers include sodium pyrosulfite, sodium edetate, erythorbic acid, magnesium oxide, and dibutylhydroxytoluene.

[0100] When preparing oral solid dosage forms, type I, type II, and / or type III crystals of compound (I) are mixed with excipients, and optionally binders, disintegrants, lubricants, colorants, flavorings, and odor-masking agents, and then tablets, coated tablets, granules, powders, capsules, etc., can be manufactured by conventional methods.

[0101] The amounts of type I, type II, and / or type III crystals of compound (I) to be included in each dosage unit are not fixed and depend on the symptoms of the patient to whom it is administered, or on the dosage form, etc. However, it is generally desirable to include approximately 0.05 to 1000 mg per dosage unit for oral preparations, approximately 0.1 to 500 mg for injectable preparations, and approximately 1 to 1000 mg for suppositories or topical preparations.

[0102] Furthermore, the daily dose of type I, type II, and / or type III crystals of compound (I) of the drug having each dosage form varies depending on the patient's symptoms, weight, age, sex, etc., and cannot be determined in general terms. However, for adults (weighing 50 kg), it is usually sufficient to administer approximately 0.05 to 5000 mg per day, preferably 0.1 to 1000 mg, and it is preferable to administer this once a day or divided into 2 to 3 doses. [Examples]

[0103] The present disclosure will be further described below with reference to examples, but the disclosure is not limited thereto. Although the present disclosure has been adequately illustrated by the examples, it will be understood that various modifications and / or modifications are possible for those skilled in the art. Accordingly, such modifications and / or modifications are incorporated into the present disclosure unless they deviate from the scope of the present disclosure.

[0104] Unless otherwise specified, commercially available reagents were used in the examples. NMR spectra were measured using an AL400 (400 MHz; JEOL (JEOL)) NMR spectrum. When tetramethylsilane was present in the deuterated solvent, tetramethylsilane was used as the internal reference; otherwise, the non-deuterated proton peak remaining in the NMR solvent was used as the internal reference, and the total δ value is expressed in ppm.

[0105] The meanings of the abbreviations are shown below. s: singlet t: triplet m: Multiplet CDCl3: Deuterated chloroform

[0106] Powder X-ray diffraction measurement Powder X-ray diffraction was performed by lightly grinding an appropriate amount of the test material in an agate mortar as needed, and then measuring it according to one of the following test conditions.

[0107] Equipment: EMPYREAN manufactured by PANalytical Reflection method (concentration method) Target: Cu X-ray tube current: 40mA X-ray tube voltage: 45kV Scanning range: 2θ = 5.0~40.0° Step: 2θ = 0.0131° Average time / step: 8.670s Scan speed: 0.0015° / s Divergence slit: 1° Scattering slit: 2.0 mm Light-receiving slit: 8.0 mm

[0108] The handling of equipment, including data processing devices, followed the methods and procedures specified for each device.

[0109] Furthermore, the numerical values ​​obtained from various spectra may vary slightly depending on the direction of crystal growth, particle size, measurement conditions, etc. Therefore, these values ​​should not be interpreted strictly.

[0110] Differential scanning calorimetry (DSC measurement) DSC measurements were performed according to the following test conditions.

[0111] Equipment: METTER TOLEDO DSC1 STAR System Sample: Approximately 1 mg Sample container: Made of aluminum Temperature range: 25~290℃ Heating rate: 10°C / min. Atmosphere gas: Nitrogen Nitrogen gas flow rate: 50 mL / min. The handling of equipment, including data processing devices, followed the methods and procedures specified for each device.

[0112] The compound 3-(15-hydroxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one, described later, is sometimes referred to as compound A.

[0113] Synthesis Example 1: Synthesis of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one (compound (I))

[0114] To a 550 mL solution of tetrahydrofuran containing 48.1 g of tert-butoxysodium, 72.9 g of 3-(15-hydroxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one in 150 mL of tetrahydrofuran was added dropwise under ice cooling and stirred for 30 minutes. 50 mL of tetrahydrofuran containing 56.8 g of methyl iodide was added to the reaction solution and stirred under ice cooling for 0.5 hours and then at room temperature for 1.5 hours. Under ice cooling, saturated ammonium chloride solution was added to the reaction solution and extracted with ethyl acetate. The mixture was washed with water and saturated brine and dried over anhydrous sodium sulfate. After filtering off the drying agent, the residue was removed by reduced-pressure distillation and purified by silka gel column chromatography (ethyl acetate / hexane) to obtain 61.5 g of the title compound.

[0115] Example 1: Preparation of type I crystals of compound (I) 1.00 g of compound (I) was mixed with 4 mL of methanol and 3 mL of water, heated and stirred, then allowed to cool to room temperature and stirred. The precipitated solid was collected by filtration and dried under reduced pressure at room temperature to obtain 892 mg of the labeled crystals. The obtained compound 1 The H-NMR spectrum was as follows: NMR(CDCl3) δppm 1.15 (s, 6H), 1.20-1.48 (m, 24H), 1.50-1.63 (m, 2H), 1.75 (s, 3H), 1.75-1.85 (m, 2H), 2.13-2.24 (m, 2H), 2.45 (t, J = 6.8 Hz, 2H), 3.33 (s, 3H), 3.36 (t, J = 6.8 Hz, 2H).

[0116] The powder X-ray diffraction spectrum (CuKα) is shown in Figure 1.

[0117] Furthermore, the characteristic diffraction angles are as follows: Characteristic diffraction angle (2θ±0.2°): 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8° (CuKα)

[0118] Furthermore, the differential scanning calorimetry (DSC) results for the Type I crystal are as follows: The differential scanning calorimetry curve is shown in Figure 2. Endothermic peak (peak top value) in differential scanning calorimetry curve: around 35°C

[0119] Example 2: Preparation of type II crystals of compound (I) 1000 mg of compound (I) was dissolved in 4 mL of acetonitrile. The mixture was cooled on ice, the precipitate was filtered off, the solid was recovered, and the resulting crystals were dried under reduced pressure at room temperature for approximately 45 minutes to obtain 747.0 mg of the indicated crystals.

[0120] The powder X-ray diffraction spectrum (CuKα) is shown in Figure 3.

[0121] Furthermore, the characteristic diffraction angles are as follows: Characteristic diffraction angle (2θ±0.2°): 6.1°, 11.4°, 12.5°, 15.1°, 19.4°, and 24.0° (CuKα)

[0122] Furthermore, the differential scanning calorimetry (DSC) results for the Type II crystal are as follows: The differential scanning calorimetry curve is shown in Figure 4. Endothermic peak (peak top value) in differential scanning calorimetry curve: around 34°C

[0123] Example 3: Preparation of Type III crystals of compound (I) 650 mg of type II crystals of compound (I) obtained in Example 2 were mixed with 3.25 mL of methanol / water mixture (9:1 v / v), and the suspension was stirred at 4°C for about 100 minutes. After that, the mixture was filtered, the solid was recovered, and it was dried under reduced pressure at room temperature for about 25 minutes. 500 mg of this solid was mixed with 2.5 mL of methanol / water mixture (9:1 v / v), and the suspension was stirred at 4°C for about 19 hours. After that, the mixture was filtered, the solid was recovered, and it was dried under reduced pressure at room temperature for about 25 minutes to obtain 335.6 mg of the labeled crystals.

[0124] The powder X-ray diffraction spectrum (CuKα) is shown in Figure 5.

[0125] Furthermore, the characteristic diffraction angles are as follows: Characteristic diffraction angle (2θ±0.2°): 11.2°, 12.4°, 14.1°, 14.9°, 18.8°, and 19.4°

[0126] Furthermore, the differential scanning calorimetry (DSC) results for the Type III crystal are as follows: The differential scanning calorimetry curve is shown in Figure 6. Endothermic peak (peak top value) in differential scanning calorimetry curve: around 34°C

[0127] Test Example 1: Solid Stability Test The solid stability of type I, type II, and type III crystals of compound (I) obtained in Examples 1-3 was evaluated after storage for 4 weeks. Storage conditions: 25℃ / 60%RH (closed system) Storage period: 4 weeks Storage amount: approx. 25mg Storage container: glass jar

[0128] Changes in the purity of compound (I) and the amount of related substances (amount of substances other than compound (I) detected) were analyzed by weighing approximately 1 mg of the sample, dissolving it in 2 mL of acetonitrile, accurately measuring 5 μL of this solution, and analyzing it by HPLC using the following method. The handling of the instruments, including data processing, followed the methods and procedures instructed for each instrument. Column: YMC-Triart C8 (4.6 x 150 mm, 3 μm) manufactured by YMC Corporation UV detection: 248nm Column temperature: 40℃ Flow rate: 1mL / min Sample cooler: 25℃ Sample concentration: 0.5 mg / mL Mobile phase A: Water / acetonitrile (1:1 v / v) mixture Mobile phase B: Acetonitrile The gradient is shown in Table 1.

[0129] [Table 1]

[0130] The results are shown in Table 2 below.

[0131] [Table 2]

[0132] As shown in Table 2, the purity of the type I, type II, and type III crystals of compound (I) showed almost no change. Furthermore, there was almost no increase in related substances in any of the type I, type II, and type III crystals of compound (I). The type I, type II, and type III crystals of compound (I) were found to be extremely stable crystals.

[0133] Test Example 2: Dynamic Moisture Absorption and Desorption Test Moisture adsorption and desorption tests were performed on type I, type II, and type III crystals of compound (I) obtained in Examples 1-3 after storage for 4 weeks. The moisture absorption and desorption test was performed according to the following conditions. Approximately 10 mg of the sample was placed in a dedicated quartz holder, and its weight at various humidity levels was continuously measured and recorded under the following conditions. The handling of the equipment, including data processing, followed the methods and procedures specified for each instrument. Equipment: VTI SA+ (manufactured by T.A. Instruments) Drying temperature: 25℃ Heating rate: 1°C / min Drying equilibrium: Ensure no decrease of 0.01 wt% in 5 minutes, within a range not exceeding 120 minutes. Measurement temperature: 25°C Humidification equilibrium: Ensure that humidity does not increase by 0.01 wt% in 5 minutes, within a range not exceeding 120 minutes. Relative humidity program: Increase humidity in 5% RH increments from 5% to 95% RH, and decrease it in 5% RH increments from 95% RH to 5% RH. The weight changes within the measurement condition range obtained in these tests are shown in Figures 7 to 9.

[0134] As shown in Figures 7-9, it was revealed that the type I, type II, and type III crystals of compound (I) exhibit almost no hygroscopicity in moisture adsorption / desorption tests at 95% relative humidity.

[0135] Therefore, the type I, type II, and type III crystals of compound (I) have low hygroscopicity, making them superior in terms of stable quality for industrial production of pharmaceuticals as candidate compounds for drug development.

[0136] Furthermore, as mentioned above, the type I, type II, and type III crystals of compound (I) are highly stable. On the other hand, by adding these crystals to water together with amphiphilic compounds such as hydroxypropyl methylcellulose and dimethyl sulfoxide (DMSO), a drug suspension or drug solution of compound (I) can be prepared.

[0137] Study Example 3: Pharmacokinetics Compound (I) was weighed in the required amount, and a drug suspension was prepared in 0.5% hydroxypropyl methylcellulose. Two rats were used in each group, and each drug suspension was orally administered to male rats (Crl:CD(SD)) using an oral gland (the dosage of the compound was 10 mg / kg). Blood was collected from the jugular vein using a syringe and needle at various blood collection points (0.5, 1, 2, 4, 8, and 24 hours after administration) (heparin sodium was used as the anticoagulant). The collected blood was centrifuged (13000 rpm, 2 min, 4°C) to prepare plasma, and after deproteinization, the compound concentration in the plasma was measured by LC / MS / MS (LCMS-8040 (shimadzu) or API4000 (AB SCIEX), LC:30-A, 20-A series (shimadzu) or Waters Acquity (waters)). Except for using compound A instead of compound (I), the drug suspension was prepared, administered, and the compound concentration was measured in the same manner as described above. The results are shown in Table 3.

[0138] As shown in Table 3, compound (I) showed a more than 1900-fold improvement in the area under the drug concentration-time curve (AUC) and an 1100-fold improvement in the maximum blood concentration (Cmax) compared to compound A. Therefore, compound (I) was shown to have unexpectedly superior pharmacokinetics compared to the known compound A.

[0139] [Table 3]

[0140] Test Example 4: Evaluation of drug efficacy in a diabetic (DM) model rat. To investigate the effects of the compound in a diabetes model, bladder function was analyzed using cystometry, referencing the method of Saitoh et al. (European Journal of Pharmacology 501(2004)143-149).

[0141] A streptozotocin (STZ) solution was prepared by dissolving STZ in citrate buffer (0.1 M citric acid, pH 4.2) to a concentration of 32.5 mg / mL. This solution was administered intraperitoneally to SD rats (65 mg / kg) to induce a DM model. The same procedure was performed on sham rats, except that citrate buffer was used instead of the STZ solution.

[0142] Blood was collected from the tail vein under awake conditions the day after model creation and four weeks later, and blood glucose levels were measured. AntSense III [Horiba, Ltd.] was used for blood glucose measurement. Individuals with blood glucose levels of 300 mg / dL or higher the day after model creation and four weeks later were judged to be successful DM models and were used in subsequent operations.

[0143] Administration of the test substances (compound A, compound (I)) began the day after the model was created. Vehicle or compound administration solution was administered orally twice daily, morning and evening, at a volume of 10 mL / kg for 4 weeks. The day after the final administration of the test substance, the rats were laparotomyed under isoflurane anesthesia and a catheter for measuring bladder pressure (PE-90, Becton Dickinson and Company) was inserted through the apex of the bladder. After closing the abdomen, the rats were placed in a restraint cage, and after waiting at least 30 minutes from the release of isoflurane anesthesia, the urethane solution was administered subcutaneously (0.8 g / kg).

[0144] A pressure transducer (DX-360, Nihon Kohden Corporation) and an infusion pump (TE-331S, Terumo Corporation) were connected to the other end of a catheter inserted into the bladder via a three-way stopcock. Bladder pressure during continuous intravesical infusion of physiological saline solution (infusion rate: 12 mL / hr) was measured using the pressure transducer and a polygraph (AP-641G, Nihon Kohden Corporation), and continuously recorded to a computer via PowerLab (ML866, AD Instruments) (sampling rate: 20 / sec). In addition, urine volume was automatically measured using an electronic balance [GX-200, A&D Corporation] set directly below the restraint cage, and continuously recorded to a computer via PowerLab (sampling rate: 20 / sec). Recording and analysis of urinary function parameters were performed using PowerLab's dedicated analysis software, Chart 5 (ver. 5.5.6, AD Instruments).

[0145] Immediately after urethane anesthesia, continuous intravesical infusion of physiological saline (infusion rate: 3 mL / hr) was performed for approximately 1.5 hours to allow the patient to acclimate to the cystometry environment. After acclimatization to the cystometry environment, urinary function (bladder pressure, urine volume, and residual urine volume) was measured by cystometry (infusion rate: 12 mL / hr).

[0146] As shown in Figure 10, it became clear that compound (I) exhibits significantly greater pharmacological efficacy compared to compound A.

[0147] Test Example 5: Measurement of Nerve Growth Promoting Effect To evaluate the nerve growth promoting effect, we analyzed the neurite bearing assay (percentage of degenerated cells) and neurite length assay (neurite length) based on the method of Topalli et al. (Brain Research 1030 (2004) 116-124).

[0148] ND3 neuronal cell lines (European Collection of Authenticated Cell Cultures) were seeded in 24-well plates in DMEM medium containing 10% fetal bovine serum (0.6 × 10⁶). 4 Cells (1 mL). After 24 hours, the culture medium of the seeded cells was removed, and a solution of compound (I), serially diluted with dimethyl sulfoxide (DMSO), was added to the cells in DMEM containing 0.1% bovine serum albumin to a final DMSO concentration of 0.1% (evaluation was performed using duplicates). Six hours after addition, the proportion of cells with protrusions longer than the diameter of the cell body (number of cells with protrusions longer than the diameter of the cell body / total number of cells) and the neurite length per cell were analyzed. The same test as above was performed, except that compound A was used instead of compound (I). The MetaMorph Neurite Outgrowth Application Module (Molecular Devices Japan Co., Ltd.) was used to analyze the total number of cells, the proportion of cells with protrusions, and the neurite length per cell. The cell number is shown as the relative number of cells, with the number of cells in the well to which DMSO was added set to 1 (Tables 4 and 5). Table 5 shows the relative activity of compound A in each test, with the maximum activity of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one (compound (I)) set to 1. Multiple tests were conducted, and the average value is shown. Data with a cell count of 0.5 or more were used. Therefore, if the cell count was less than 0.5, the protrusion length per cell and the percentage of cells with protrusions are indicated as "-".

[0149] As shown in Table 4, compound (I) resulted in a relative cell count of 0.5 or higher when added at concentrations of 1 to 100 μM.

[0150] [Table 4]

[0151] As shown in Table 5, compound (I) was confirmed to have superior nerve growth-promoting effects compared to compound A.

[0152] Table 5

Claims

1. Crystals of 3-(15-methoxypentadecyl)-2,4,4-trimethylcyclohexa-2-en-1-one having at least five peaks with diffraction angles (2θ ± 0.2°) selected from 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8° in powder X-ray diffraction spectrum (CuKα).

2. The crystal according to claim 1, wherein the powder X-ray diffraction spectrum (CuKα) has peaks at diffraction angles (2θ ± 0.2°) of 7.0°, 14.0°, 17.5°, 19.5°, 21.0°, 23.7°, and 24.8°.

3. The crystal according to claim 1 or 2, wherein the powder X-ray diffraction spectrum is the same as that shown in Figure 1 below. 。

4. A crystal according to any one of claims 1 to 3, having an endothermic peak with a peak temperature of 33°C to 37°C in differential scanning calorimetry (DSC measurement).

5. A pharmaceutical composition containing the crystal described in any one of claims 1 to 4.

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