Crystals of a boronic acid derivative, a method for producing the same, and use thereof

Abstract

The present disclosure relates to crystals of boric acid derivatives, a method for producing the same, and use thereof. Specifically, the present disclosure provides crystals of a compound of Formula I, a method for producing the same, and use thereof. JPEG2025522567000021.jpg3864

Description

【Technical Field】 【0001】 The present disclosure belongs to the pharmaceutical field and relates to a crystal of a boronic acid derivative, a method for producing the same, and its use. 【Background Art】 【0002】 Phosphodiesterase (PDE) is a type of intracellular enzyme that cleaves the phosphodiester bond of 3′,5′-cyclic adenosine monophosphate (cAMP) and 3′,5′-cyclic guanosine monophosphate (cGMP) as second messenger molecules. Cyclic nucleotides cAMP and cGMP act as second messengers in various cell pathways. Among them, PDE4 is highly specific for cAMP and has four subtypes: PDE4A, PDE4B, PDE4C, and PDE4D. PDE4 is involved in the activation of monocytes and macrophages, neutrophil infiltration, vascular smooth muscle proliferation, vasodilation and myocardial contraction, and the promotion of other related physiological and pathological processes, and affects central nervous system function, cardiovascular function, inflammatory / immune system, cell adhesion, etc. PDE4 plays a major role in regulating the expression of inflammatory and anti-inflammatory mediators, and PDE4 inhibitors can inhibit the release of harmful mediators by inflammatory cells. 【0003】 In recent years, many PDE4 inhibitors have been identified. For example, roflumilast has been approved to reduce the frequency of sudden exacerbations of severe chronic obstructive pulmonary disease (COPD) and prevent the exacerbation of COPD symptoms, and apremilast has been approved as a therapeutic agent for adults with active psoriatic arthritis. Although PDE4 inhibitors exhibit excellent pharmacological activity, these PDE inhibitors have side effects such as inducing gastrointestinal symptoms such as vomiting and diarrhea. The development of selective PDE4 inhibitors, particularly selective PDE4 inhibitors with affinity for PDE4B and PDE4D, is still needed. 【0004】 Crisaborole, a boron (B)-containing agent, was approved by the FDA on December 14, 2016, as a topical treatment for mild to moderate atopic dermatitis. Boron atoms promote penetration into the skin and bind to the two-metal center of phosphodiesterase 4 (PDE4). In addition, other small molecule boron-containing PDE inhibitors have been reported, such as Chinese Patent Application Publication No. 102014927 and International Publication No. 2020070651. However, the compounds of the present disclosure are not disclosed in any literature, and such compounds exhibit a specific PDE4 inhibitory effect. 【0005】 PCT / CN2021 / 141010 provides a novel PDE4 inhibitor named (R)-4-(6-(8-methoxy-2,2-dimethylbenzodihydropyran-5-yl)pyrazin-2-yl)-1,2-oxaborolane-2-ol having the structure shown in Formula I: 【Chemical formula】 【0006】 The crystalline form of a pharmaceutical active ingredient often affects the chemical stability of the drug. When the crystallization conditions and storage conditions are different, the crystal structure of the compound may change, and other crystal forms may be generated. Generally, amorphous pharmaceuticals having no regular crystal structure often have other defects such as poor product stability, too fine crystals, difficult filtration, easy aggregation, and poor fluidity. The polymorphs of a drug have different requirements for the storage, manufacture, and amplification of the product. Therefore, in order to improve various properties of the above-mentioned compound, it is necessary to conduct a detailed study on the crystal form of the above-mentioned compound. 【Summary of the Invention】 【0007】 The present disclosure provides a crystalline form A of a compound of Formula I, characterized in that the X-ray powder diffraction spectrum represented by the diffraction angle 2θ has characteristic peaks at 7.537°, 15.219°, 21.798°, and 22.911°: 【Chemical formula】 ​ 【0008】 In some embodiments, the crystalline form A of the compound of formula I has characteristic peaks at 7.537°, 11.366°, 14.297°, 15.219°, 16.190°, 17.623°, 19.134°, 21.798°, 22.911° and 24.735° in the X-ray powder diffraction spectrum represented by the diffraction angle 2θ. 【0009】 In some embodiments, the crystalline form A of the compound of formula I has characteristic peaks at 7.537°, 11.366°, 14.018°, 14.297°, 15.219°, 16.190°, 17.623°, 19.134°, 21.062°, 21.798°, 22.911°, 24.735°, 25.123° and 26.884° in the X-ray powder diffraction spectrum represented by the diffraction angle 2θ. 【0010】 In some embodiments, the crystalline form A of the compound of formula I has characteristic peaks at 7.537°, 11.366°, 14.018°, 14.297°, 15.219°, 16.190°, 17.623°, 18.412°, 19.134°, 20.010°, 21.062°, 21.798°, 22.911°, 23.889°, 24.735°, 25.123°, 26.884°, 28.343°, 29.645°, 33.499° and 36.362° in the X-ray powder diffraction spectrum represented by the diffraction angle 2θ. 【0011】 In some embodiments, the crystalline form A of the compound of formula I has an X-ray powder diffraction spectrum represented by the diffraction angle 2θ as shown in FIG. 7. 【0012】 In some embodiments, the crystalline form A of the compound of formula I of the present disclosure has an error range of ±0.2° for the 2θ angle. 【0013】 The present disclosure also provides a method for producing the crystalline form A of the compound of formula (I) above, and the production method is Method I or Method II: Method I is: (a1) Formula I: 【Chemical formula】 The step of mixing the compound of with solvent (1); (b1) The step of crystallizing under stirring; comprising wherein the solvent (1) is one or more selected from the group consisting of water, nitromethane, N,N-dimethylformamide (DMF), acetonitrile, methanol, ethanol, acetone, isopropanol (IPA), methyl ethyl ketone (MEK), tetrahydrofuran (THF), 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether (MTBE), 1,2-xylene, toluene, dioxane and hexane; 【0014】 Method II comprises: (a2) The step of mixing the compound of formula I with solvent (2); (b2) The step of crystallizing under volatilization; comprising wherein the solvent (2) is one or more selected from the group consisting of acetone, acetonitrile, dichloromethane (DCM), dioxane, ethanol, ethyl acetate, heptane, isopropanol, methanol, methyl ethyl ketone, methyl tert-butyl ether, toluene and water. 【0015】 In some embodiments, the solvent (1) is water, nitromethane, N,N-dimethylformamide, acetonitrile, ethanol, acetone, isopropanol, methyl ethyl ketone, tetrahydrofuran, 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether, 1,2-xylene, toluene, dioxane, hexane or a mixed solvent of water and methanol. 【0016】 In some embodiments, when the solvent (1) is a mixed solvent of water and methanol, the molar ratio of water to methanol is 0.37 to 0.90, and in non-limiting embodiments, it may be 0.37, 0.40, 0.47, 0.50, 0.57, 0.60, 0.66, 0.70, 0.74, 0.80, 0.82 and 0.90. 【0017】 In some embodiments, the temperature of crystallization under stirring is from 10°C to 60°C, and in non-limiting embodiments, it may be 10°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, and 60°C. 【0018】 In some embodiments, the temperature of crystallization under stirring is room temperature. 【0019】 In some embodiments, when the temperature of crystallization under stirring is from 10°C to 40°C (e.g., room temperature), the solvent (1) is water, nitromethane, acetonitrile, ethanol, acetone, isopropanol, methyl ethyl ketone, tetrahydrofuran, 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether, 1,2-xylene, toluene, dioxane, hexane, or a mixed solvent of water and methanol. 【0020】 In some embodiments, when the temperature of crystallization under stirring is from 41°C to 60°C (e.g., 50°C), the solvent (1) is water, N,N-dimethylformamide, acetonitrile, acetone, methyl ethyl ketone, 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether, 1,2-xylene, toluene, hexane, or a mixed solvent of water and methanol. 【0021】 In some embodiments, the crystallization under stirring is crystallization by pulping. 【0022】 In some embodiments, the solvent (2) is acetone, acetonitrile, dichloromethane, dioxane, ethyl acetate, heptane, isopropanol, methyl ethyl ketone, methyl tert-butyl ether, toluene, water, or the following mixed solvents: acetone and acetonitrile, acetone and dichloromethane, acetone and dioxane, acetone and ethanol, acetone and ethyl acetate, acetone and heptane, acetone and isopropanol, acetone and methanol, acetone and methyl ethyl ketone, acetone and methyl tert-butyl ether, acetone and tetrahydrofuran, acetone and toluene, acetone and water, acetonitrile and dichloromethane, acetonitrile and dioxane, acetonitrile and ethanol, acetonitrile and ethyl acetate, acetonitrile and isopropanol, acetonitrile and methyl ethyl ketone, acetonitrile and methyl tert-butyl ether, acetonitrile and tetrahydrofuran, acetonitrile and toluene, dichloromethane and dioxane, dichloromethane and ethanol, dichloromethane and ethyl acetate, dichloromethane and heptane, dichloromethane and isopropanol, dichloromethane and methyl ethyl ketone, dichloromethane and methyl tert-butyl ether, dichloromethane and tetrahydrofuran, dichloromethane and toluene, dioxane and methyl ethyl ketone, dioxane and tetrahydrofuran, dioxane and toluene, ethanol and ethyl acetate, ethanol and methanol, ethanol and tetrahydrofuran, ethanol and toluene, ethanol and water, ethyl acetate and heptane, ethyl acetate and isopropanol, ethyl acetate and methanol, ethyl acetate and methyl ethyl ketone, ethyl acetate and methyl tert-butyl ether, ethyl acetate and toluene, heptane and methyl ethyl ketone, heptane and methyl tert-butyl ether, heptane and tetrahydrofuran, heptane and toluene, methanol and methyl ethyl ketone, methanol and methyl tert-butyl ether, methanol and toluene, methanol and water, methyl ethyl ketone and methyl tert-butyl ether, methyl ethyl ketone and tetrahydrofuran, methyl ethyl ketone and toluene, methyl tert-butyl ether and tetrahydrofuran, methyl tert-butyl ether and toluene, tetrahydrofuran and toluene,It is either a group of tetrahydrofuran or water. 【0023】 In some embodiments, when the solvent (2) is a mixed solvent, the volume ratio of the two is 1:3 to 3:1, preferably 1:1. 【0024】 In some embodiments, the temperature of crystallization by volatilization is 10°C to 40°C, and in non-limiting embodiments, it may be 10°C, 20°C, 25°C, 30°C, 35°C, and 40°C. 【0025】 In some embodiments, the temperature of crystallization by volatilization is room temperature. 【0026】 In some embodiments, the volume (μl) of the solvent (1) or (2) may be 1 to 200 times the mass (mg) of the compound of formula I, and in non-limiting embodiments, it may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 200 times or any value between the two numbers. 【0027】 In some embodiments, after crystallization under stirring or crystallization under volatilization, it further includes the steps of filtration, washing, or drying. 【0028】 The present disclosure also provides a pharmaceutical composition comprising the crystalline form A or the crystalline form A produced by the manufacturing method described above, and optionally a pharmaceutically acceptable excipient. 【0029】 The present disclosure also provides a method for manufacturing the pharmaceutical composition, which includes the step of mixing the crystalline form A or the crystalline form A produced by the method with a pharmaceutically acceptable excipient. 【0030】 The present disclosure also provides the use of the crystalline form A, or the crystalline form A produced by the production method, or the pharmaceutical composition in the manufacture of a medicament for the prevention and / or treatment of PDE-related diseases. In some embodiments, the PDE-related disease is asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes, allergic rhinitis, allergic conjunctivitis, ulcerative colitis, or rheumatism. 【0031】 The present disclosure also provides the use of the crystalline form A, or the crystalline form A produced by the production method, or the pharmaceutical composition in the manufacture of a medicament for the prevention and / or treatment of asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes, allergic rhinitis, allergic conjunctivitis, ulcerative colitis or rheumatism. 【0032】 The term "2θ or 2θ angle" in the present disclosure refers to the diffraction angle, where θ is the Bragg angle, and its unit is ° or degree. The error range of 2θ for each characteristic peak is ±0.2 (including the case of rounding to the first decimal place), which is -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19 or 0.20. 【0033】 In the "9103 Guidelines for Hygroscopicity of Medicaments" in Volume 4 of the Chinese Pharmacopoeia 2015 Edition, the hygroscopic characteristics are described, and the weight increase due to moisture absorption is defined as follows. Deliquescence: Absorbing sufficient moisture to form a liquid; Very highly hygroscopic: The weight increase due to moisture absorption is 15% or more; Hygroscopic: The weight increase due to moisture absorption is less than 15% but 2% or more; Slightly hygroscopic: The weight increase due to moisture absorption is less than 2% but 0.2% or more; Non-hygroscopic or almost non-hygroscopic: The weight increase due to moisture absorption is less than 0.2%. 【0034】 In the present disclosure, the term "differential scanning calorimetry (DSC)" means measuring the temperature difference and heat flow difference between a sample and a reference while the temperature of the sample is rising or constant, characterizing all physical and chemical changes related to the heat effect, and obtaining phase change information of the sample. 【0035】 The drying temperature in the present disclosure is usually in the range of 25°C to 100°C, preferably 40°C to 70°C. Drying can be carried out under normal pressure or under reduced pressure of less than -0.08 MPa. 【0036】 "Room temperature" in the present disclosure is 25°C ± 5°C. 【0037】 The term "excipient" in the present disclosure includes, but is not limited to, adjuvants, carriers, glidants, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents or emulsifiers approved by the US Food and Drug Administration for use in humans or livestock. 【0038】 The term "pulping" in the present disclosure refers to a purification method that utilizes the property that the compound has low solubility in the solvent but the impurities have high solubility in the solvent. By pulping purification, color can be removed, crystal form can be changed, and trace amounts of impurities can be removed. 【0039】 The starting materials used in the method for producing the crystalline form of the present disclosure can be compounds in any form, including, but not limited to, amorphous forms, any crystalline forms, hydrates, solvates, etc. 【0040】 In the present disclosure, values related to the determination and calculation of the substance content necessarily have a certain degree of error. Generally, ±10% belongs to a reasonable error range. The degree of error varies depending on the situation in which it is used, and the error may be ±10% or less, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2% or ±1%, preferably ±5%. 【0041】 In the present disclosure, the compound of formula I (R)-4-(6-(8-methoxy-2,2-dimethylbenzo[d][1,3]dioxol-5-yl)pyrazin-2-yl)-1,2-oxaborolane-2-ol (amorphous, hereinafter referred to as compound I) is produced according to the method described in PCT / CN2021 / 141010, and the related content thereof is incorporated herein by reference for the purpose of explanation. 【Brief Description of the Drawings】 【0042】 【Figure 1】 Figure 1 shows the XRPD spectrum of amorphous compound I. 【Figure 2】 Figure 2 shows a graph comparing the clinical scores of the compounds in each group for the disease model in Test Example 4. 【Figure 3】 Figure 3 shows a graph comparing the clinical scores of the compounds in each group for the erythema disease model in Test Example 4. 【Figure 4】 Figure 4 shows a graph comparing the clinical scores of the compounds in each group for the psoriasis disease model in Test Example 4. 【Figure 5】 Figure 5 shows a graph comparing the inhibitory effects of the compounds in each group on the increase in skin thickness in Test Example 4. 【Figure 6】 Figure 6 shows a graph comparing the effects of the compounds in each group on the spleen weight to body weight ratio in Test Example 4. 【Figure 7】 Figure 7 shows the XRPD spectrum of crystalline form A. 【Figure 8】 Figure 8 shows the DSC spectrum of crystalline form A. 【Figure 9】 Figure 9 shows the TGA spectrum of crystalline form A. 【Figure 10】Figure 10 shows the DVS spectrum of crystalline form A. In Figures 1 to 6, * indicates p < 0.05 compared with the model group, ** indicates p < 0.01 compared with the model group, and *** indicates p < 0.001 compared with the model group. The symbols ▼, ●, ◆, and # are used only to distinguish groups and have no actual meaning. 【Modes for Carrying Out the Invention】 【0043】 Hereinafter, the present disclosure will be described in more detail through examples. The examples or experimental examples of the present disclosure are only intended to explain the technical solutions of the present disclosure and do not limit the spirit and scope of the present disclosure. 【0044】 The abbreviations used in the present disclosure are explained below. XRPD X-ray powder diffraction DSC Differential scanning calorimetry TGA Thermogravimetric analysis DVS Dynamic vapor sorption 1 H-NMR Solution nuclear magnetic resonance hydrogen spectroscopy DMF N,N-Dimethylformamide MEK Methyl ethyl ketone MTBE Methyl tert-butyl ether THF Tetrahydrofuran IPA Isopropanol DCM Dichloromethane ACN Acetonitrile 【0045】 Test conditions of the equipment used in the experiments of the present disclosure: 1. X-ray powder diffraction, XRPD Equipment model: Malver Panalytical Aeris X-ray powder diffractometer Radiation: Monochromatic Cu-Kα ray (λ = 1.54188) Scan mode: θ / 2θ, scan range (2θ range): 3.5 - 50° Voltage: 40 kV, current: 15 mA 【0046】 2. Differential scanning calorimetry, DSC Machine model: TA DSC250 Purge gas: nitrogen; Nitrogen purge rate: 50 mL / min Heating rate: 10 °C / min Temperature range: 25 °C to 250 °C 【0047】 3. Thermogravimetric analysis, TGA Machine model: TA TGA550 Purge gas: nitrogen; Nitrogen purge rate: 20 ml / min Heating rate: 10 °C / min Temperature range: 30 °C to 350 °C 【0048】 4. Dynamic vapor sorption (DVS) Using SMS Intrinsic PLUS, at 25 °C, with a humidity change of 50% - 0% - 90% and a step width of 10%, detection is performed, and the judgment criteria are that the mass change dM / dT of each gradient is less than 0.0002%, TMAX is 360 minutes, and 2 cycles are executed. 【0049】 5. The high-performance liquid chromatography (HPLC) described in the stability test of crystalline form A of the present disclosure is measured with an Agilent 1260 Infinity II under the following HPLC conditions; Agilen XDB C18 5μm 4.6×250mm chromatography column; mobile phase, A - 0.05% TFA (in water) and B - ACN; flow rate 1.0 ml / min, wavelength 220 nm. 【0050】 6. The structure of the compound is identified by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The NMR shift (δ) is expressed in 10 -6 (ppm) units. NMR is measured with a Bruker AVANCE-400 nuclear magnetic spectrometer. The measurement solvents are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (methanol-d4), and the internal standard is tetramethylsilane (TMS). 【0051】 7. The HPLC in the production example of Compound I was measured with an Agilent 1100 high-pressure liquid chromatograph equipped with a GAS15B DAD UV detector and a Waters Vbridge C18 150×4.6 mm 5 μm column. 【0052】 8. MS was measured with an Agilent 6120 triple quadrupole mass spectrometer equipped with a G1315D DAD detector and a Waters Xbridge C18 4.6×50 mm, 5 μM column, and scanned in positive / negative ion mode with a mass scan range of 80 - 1200. 【0053】 9. Preparative HPLC conditions: water; Sunfire column (Prep C18 OBD 19×250 mm 10 μm). 【0054】 10. Chiral column separation conditions: Chiralpak IG column 5 μm 30×250 mm; mobile phase Hex:EtOH = 35:65, 15 mL / min; temperature 30 °C; wavelength 254 nm. 【0055】 11. For thin-layer chromatography, a Yantai Huanghai HSGF254 silica gel plate is used. The specifications of the gel plate used are 0.2 mm ± 0.03 mm for thin-layer chromatography (TLC) and 0.4 mm - 0.5 mm for separation and purification of products by thin-layer chromatography. 【0056】 12. For the high-speed column purification system, Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage) is used. 【0057】 13. As the carrier for normal-phase column chromatography, Yantai Huanghai 200 - 300 mesh silica gel or 300 - 400 mesh silica gel is generally used. Alternatively, a Changzhou Santai pre-packed ultra-high purity normal-phase silica gel column (40 - 63 μm, 60 g, 24 g, 40 g, 120 g or other specifications) is used. 【0058】 Known starting materials of the present disclosure can be synthesized by or according to methods known in the art, or can be purchased from Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Bide Pharm and other companies. 【0059】 Unless otherwise specified in the examples, all reactions can be carried out under a nitrogen atmosphere. 【0060】 A nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon with a volume of about 1 L. 【0061】 A hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1 L. 【0062】 Hydrogen is generated by a QPH-1L type hydrogen generator from Shanghai Quan Pu Scientific Instruments. 【0063】 A nitrogen atmosphere or a hydrogen atmosphere is usually generated by repeating evacuation and filling with nitrogen or hydrogen three times. 【0064】 Unless otherwise specified in the examples, the solution refers to an aqueous solution. 【0065】 Unless otherwise specified in the examples, the reaction temperature is room temperature of 20°C to 30°C. 【0066】 The reaction process in the examples is traced by thin layer chromatography (TLC). The developing agent used in the reaction, the eluent system of column chromatography used for purification of the compound, the developing system of thin layer chromatography and the volume ratio of the solvents are adjusted according to the polarity of the compound. It can also be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid. 【0067】 Example 1: Preparation of Compound I 【Chem.】 【Chem.】 【0068】 Step 1): Compound 1a (2.0 g, 14.6 mmol) and triethylamine (1.8 g, 17.8 mmol) were dissolved in N,N-dimethylformamide (30 mL), and the solution was cooled to 0 °C. tert-Butyldiphenylchlorosilane (4.0 g, 14.6 mmol) was added dropwise to the reaction system under a nitrogen atmosphere. The reaction system was warmed to room temperature and continuously stirred until the completion of the reaction was detected by TLC. The reaction solution was poured into water and extracted with ethyl acetate (100 mL × 3). The organic phase was washed with water (50 mL × 2), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether) to obtain Compound 1b (5.1 g), which was used directly in the next reaction. 【0069】 Step 2): A mixture of Compound 1b (5.1 g, 13.6 mmol), bis(pinacolato)diboron (4.2 g, 16.3 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (512 mg, 0.7 mmol), and potassium acetate (2.0 g, 20.4 mmol) in dioxane (100 mL) was warmed to 80 °C and stirred overnight under a nitrogen atmosphere. The reaction solution was poured into water and extracted with ethyl acetate (100 mL × 2). The organic phase was washed with water (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate) to obtain Compound 1c (2.0 g). LCMS: m / z 423.2 (M+H) + . 【0070】 Step 3): Compound a (100 g, 492 mmol), DMF (1000 mL), potassium iodide (92.6 g, 837 mmol), copper(I) iodide (3.13 g, 9.85 mmol), and potassium carbonate (136 g, 985 mmol) were successively added to a 2000 mL one-neck flask at room temperature. 3-Chloro-3-methyl-1-butyne (100 mL, 886 mmol) was added dropwise under a nitrogen atmosphere. The reaction mixture was stirred at 70 °C for 16 h. The resulting mixture was cooled to room temperature. Water (1000 mL) was added, and the mixture was extracted with petroleum ether (1000 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate) to obtain compound b (50 g). 1 1H NMR (400 MHz, CDCl3) δ 7.57 (d, J = 2.4 Hz, 1H), 7.15 (dd, J = 8.8, 2.4 Hz, 1H), 6.76 (d, J = 8.8 Hz, 1H), 3.79 (s, 3H), 2.58 (s, 1H), 1.65 (s, 6H). 【0071】 Step 4): Compound b (20.0 g, 74.4 mmol), n-hexane (200 mL), and calcium carbonate palladium (1.95 g, 18.8 mmol) were successively added to a 500 mL one-neck flask at room temperature. The reaction mixture was stirred at room temperature for 16 h under a hydrogen atmosphere. The resulting mixture was filtered and concentrated under reduced pressure to obtain compound c (19 g). 1 1H NMR (400 MHz, CDCl3) δ 7.15 (d, J = 2.4 Hz, 1H), 7.09 (dd, J = 8.8, 2.4 Hz, 1H), 6.73 (d, J = 8.8 Hz, 1H), 6.12 (dd, J = 17.6, 10.8 Hz, 1H), 5.14 (dd, J = 20.0, 9.2 Hz, 2H), 3.79 (s, 3H), 1.46 (s, 6H). 【0072】 Step 5): Compound c (10.0 g, 36.9 mmol) and diethylaniline (10 mL, 62.5 mmol) were sequentially added to a 50 mL one-neck flask at room temperature. The reaction mixture was stirred at 210 °C for 1 hour. The resulting mixture was cooled to room temperature, adjusted to neutral pH with 1 M HCl, extracted with ethyl acetate (200 mL × 3), and washed with water (200 mL × 3). The organic phase was concentrated under reduced pressure to obtain Compound d (9.1 g). 1 1H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 6.97 (d, J = 8.8 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 5.14 - 5.01 (m, 1H), 3.78 (s, 3H), 3.40 (d, J = 6.8 Hz, 2H), 1.74 (s, 3H), 1.63 (s, 3H). 【0073】 Step 6): Compound d (5.00 g, 18.5 mmol), toluene (25 mL) and Amberlyst® 15 (5.00 g, 15.9 mmol) were sequentially added to a 100 mL one-neck flask at room temperature. The reaction mixture was stirred at 100 °C for 2 hours under a nitrogen atmosphere. The resulting mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure to obtain Compound e (3.89 g). 1 1H NMR (400 MHz, DMSO-d6) δ 7.04 (d, J = 8.8 Hz, 1H), 6.75 (d, J = 8.8 Hz, 1H), 3.71 (s, 3H), 2.63 (t, J = 6.8 Hz, 2H), 1.78 (t, J = 6.8 Hz, 2H), 1.26 (s, 6H). 【0074】 Step 7): Compound e (28.4 g, 105 mmol), dioxane (300 mL), bis(pinacolato)diboron (31.9 g, 126 mmol), potassium acetate (20.6 g, 209 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (7.66 g, 10.5 mmol) were sequentially added to a 500 mL one-neck flask at room temperature. The reaction mixture was stirred at 105 °C for 16 hours under a nitrogen atmosphere. The resulting mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate) to obtain compound f (28.4 g). LCMS: m / z 319 (M+H) + 。 【0075】 Step 8): Compound f (6.20 g, 19.5 mmol), 1,4-dioxane (80 mL), water (16 mL), 2,6-dichloropyrazine (2.90 g, 39.0 mmol), potassium carbonate (5.39 g, 39.0 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.43 g, 1.95 mmol) were sequentially added to a 250 mL one-neck flask at room temperature. The reaction mixture was stirred at 110 °C for 16 hours under a nitrogen atmosphere. The resulting mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate) to obtain compound g (5.7 g). LCMS: m / z 305 (M+H) + 。 【0076】 Step 9): Compound g (35.0 g, 115 mmol), Compound 1c (44.5 g, 149 mmol), potassium carbonate (23.8 g, 172 mmol), potassium acetate (16.9 g, 172 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (8.41 g, 11.5 mmol) were added to a mixed solvent of 1,4-dioxane (250 mL) and water (5 mL) in a 500 mL one-neck flask at room temperature. The reaction mixture was stirred until dissolved, purged with nitrogen three times, and stirred at 110 °C for 16 h. The resulting mixture was cooled to room temperature, extracted with ethyl acetate (300 mL × 3), and dried over anhydrous sodium sulfate. The organic phase was filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate) to obtain Compound h (37 g). LCMS: m / z 441.1 (M+H) + 。 【0077】 Step 10): 1,2-Bis(diphenylphosphino)ethane (1.81 g, 4.54 mmol), (1,5-cyclooctadienyl)methoxyiridium(I) dimer (1.50 g, 2.27 mmol), and anhydrous 1,2-dichloroethane (100 mL) were added to a 250 mL three-neck flask at room temperature. The mixture was stirred at room temperature for 10 min, and Compound h (10.0 g, 22.7 mmol) was added. The mixture was warmed to 70 °C, and pinacol borane (20.3 g, 159 mmol) was added dropwise. The reaction was carried out at 95 °C for 4 h. The resulting mixture was cooled to 0 °C, and methanol (50 mL) was added dropwise to stop the reaction. The resulting mixture was concentrated, and the residue was purified by column chromatography (petroleum ether / ethyl acetate) to obtain Compound i (1.4 g). LCMS: m / z 569.3 (M+H) + 。 【0078】 Step 11): Compound i (9.00 g, 15.8 mmol) and tetrahydrofuran (30 mL) were added to a 50 mL one-neck flask at room temperature. The mixture was cooled to 0 °C, and 2 M hydrochloric acid (50 mL) was added. The reaction mixture was stirred at 50 °C for 16 h. The resulting mixture was concentrated under reduced pressure, extracted with ethyl acetate (300 mL × 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (acetonitrile / water / trifluoroacetic acid) to obtain Compound I-A (3.82 g). 【0079】 The product was subjected to chiral separation (column: Chiralpak IG 5 μm 30 × 250 mm, mobile phase: Hex:EtOH = 35:65; flow rate: 15 mL / min; temperature: 30 °C; wavelength: 254 nm) to obtain Compound I (with a shorter retention time). 【0080】 Compound I Compound I was characterized by XRPD, and as a result, it was shown that the product was amorphous. The XRPD spectrum is shown in Figure 1. LCMS: m / z 355.0 (M+H) + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 1H), 8.48 (s, 1H), 6.98 (d, J = 8.4 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 4.29 (dd, J = 8.8, 7.6 Hz, 1H), 3.96 (dd, J = 8.8, 6.8 Hz, 1H), 3.77 (s, 3H), 3.74 - 3.62 (m, 1H), 2.93 - 2.67 (m, 2H), 1.68 (t, J = 6.8 Hz, 2H), 1.38 - 1.24 (m, 7H), 1.21 - 1.08 (m, 1H). 【0081】 Example 2: Preparation of Crystal Form A of Compound I 10 mg of Compound I was added to 0.2 ml of the solvent as shown in Table 1 below. The compound was kept in a slurry state in the solvent and stirred at room temperature (25 °C) or 50 °C for 7 days. The solvent was quickly removed using filter paper, and the resulting solid was determined to be Crystal Form A by XRPD. 【0082】 【Table 1】 【0083】 Example 3: Preparation of Crystal Form A of Compound I 5 mg of Compound I was added to the solvent as shown in Table 2 below. Evaporation was carried out at room temperature. After 7 days, the resulting mixture was placed in a vacuum drying oven and the solvent was removed at 50 °C. The resulting solid was determined to be Crystal Form A by XRPD. 【0084】 【Table 2】 【0085】 The solids obtained in Examples 3 and 4 were detected by X-ray powder diffraction, and the product was defined as Crystal Form A. The XRPD spectrum is shown in Figure 7, and the positions of the characteristic peaks are shown in Table 3. The DSC spectrum shown in Figure 8 shows an endothermic peak at 140.43 °C. The TGA spectrum shown in Figure 8 shows a weight loss of 1.53% between 25 °C and 100 °C. 【0086】 As shown in Figure 10, DVS detection shows that the weight increase due to moisture absorption of the sample is about 0.08% under normal storage conditions (i.e., 25 °C, 60% RH), about 0.15% under accelerated experimental conditions (i.e., 70% RH), and about 0.70% under extreme conditions (i.e., 90% RH). During the change of humidity from 0% to 90% RH, the desorption process of the sample coincides with its adsorption process. After DVS detection, the crystal form was redetermined, and XRPD detection shows that the crystal form does not change before and after DVS detection. 【0087】 【Table 3】 【0088】 Example 4: Stability Test I of Crystal Form A Determine whether temperature and humidity affect the stability of crystalline form A of the compound of formula I. 【0089】 Crystalline form A was placed in a stability chamber and an accelerated stability test was conducted under a series of temperature and humidity combinations. After being placed for different days under various temperature and humidity conditions, the samples were taken out and compared with the initial crystalline form A. The content of the compound of formula I was measured by high performance liquid chromatography and XRPD detection was performed. The test results are shown in Table 4 below. 【0090】 The results indicate that crystalline form A remains unchanged and has good stability. 【0091】 [Table 4] 【0092】 Example 5: Stability Test II of Crystalline Form A Determine whether light affects the stability of crystalline form A of the compound of formula I. 【0093】 Crystalline form A was placed in a light stability chamber and taken out after reaching the ICH intensity. The obtained solid was compared with the initial crystalline form A. The content of the compound of formula I was determined by high performance liquid chromatography and XRPD detection was performed. The test results are shown in Table 5 below. 【0094】 [Table 5] 【0095】 Biological evaluation Hereinafter, the present disclosure will be further described with reference to test examples, but these test examples do not limit the scope of the present disclosure. 【0096】 Structure of Compound A: [Chemical Formula] 【0097】 Compound A was produced by the method disclosed in Example 4 on page 181 of the specification of International Publication No. 2020070651 of the patent application. 【0098】 Test Example 1. In vitro PDE4B Enzyme Activity Detection Assay 1. Experimental Materials 【Table 6】 【0099】 2. Experimental Procedures A stock solution of the compound at a concentration of 10 mM was prepared in vitro using 90% DMSO (10% water), and using this, a series of dilutions with a dilution factor of 1:5, starting from a final concentration of 100 μM and decreasing to 0.05 nM, were prepared. 【0100】 0.2 μl of the compound solution was transferred to a 384-well reaction plate, and 0.2 μl of 100% DMSO was transferred to both the negative control and the positive control. Next, 10 μl of 2×PDE4B1 enzyme solution (final concentration 0.04 nM) was added to the wells, and for the control wells without enzyme activity, 10 μl of 1× reaction buffer (50 mM HEPES, pH 7.5, 0.0015% Brij-35) was added instead of the enzyme solution. The plate was centrifuged at 1,000 rpm for 1 minute and incubated at room temperature for 15 minutes. Next, 10 μl of 2× FAM-cAMP substrate solution (final substrate concentration 0.1 μM) was added to each well of the 384-well reaction plate. The plate was centrifuged at 1,000 rpm for 1 minute and incubated at room temperature for 15 minutes. Next, 10 μl of 2× FAM-cAMP substrate solution (final substrate concentration 0.1 μM) was added to each well of the 384-well reaction plate. The plate was centrifuged at 1,000 rpm for 1 minute and the reaction was carried out at 25°C for 30 minutes. After the reaction was completed, 60 μL of reaction stop solution was added to each well of the 384-well reaction plate to stop the reaction, and it was incubated in the dark at room temperature for 60 minutes while shaking at 600 rpm on a shaker. 【0101】 After the incubation was completed, the RLU data was read and the inhibition rate was calculated. IC50 The values are calculated from the concentration inhibition approximation curve, with the maximum value referring to the reading of the DMSO control and the minimum value referring to the reading of the control without enzyme activity. 【0102】 The in vitro inhibition of PDE4B1 enzyme activity by Compound I of the present disclosure was determined by the said test, and the measured IC 50 values are shown in Table 6. 【0103】 [Table 7] 【0104】 Test Example 2. Inhibitory effect of the compound on the release of inflammatory cytokines in peripheral blood mononuclear cells (PBMCs) The frozen PBMCs were thawed, and the cell viability and cell number were detected by trypan blue staining. The thawed PBMCs were washed with RPMI1640 complete medium (RPMI1640 + 10% FBS + 1% PS), centrifuged, and the supernatant was discarded. The PBMCs were resuspended in RPMI1640 complete medium, and the cell density was adjusted to 2×10 6 cells / mL. 2×10 5 The PBMC cells were seeded in a 96-well cell culture plate, and the compound to be tested was added at different concentrations in two wells each at 9-step concentrations with a 1:5 serial dilution starting from the maximum compound concentration of 100 μM. LPS was added at a final concentration of 0.1 ng / mL to make the total volume 200 μL. For the negative control and positive control, only LPS and DMSO were added to the negative control wells, and in addition to cells and LPS, 1 μg / mL of dexamethasone was added as a positive control to the positive control wells. The cells were incubated in an incubator at 37 °C for 24 hours. After the incubation, 100 μL of the cell culture supernatant was collected, and the TNF-α level was detected by ELISA. 100 μL of CellTiter-Glo was added to the remaining cells in each well to detect the cell viability. The IC 50 values of TNF-α release inhibition by the compound were calculated. 【0105】 The in vitro inhibition of the release of inflammatory cytokines in PBMC of Compound I of the present disclosure was determined by the said test, and the measured IC 50 values are shown in Table 7. 【0106】 [Table 8] 【0107】 Test Example 3. In vitro inhibition of the secretion of IL-23 by a compound in DC cells differentiated from human monocytes Day 0: Monocytes were isolated and purified from fresh human peripheral blood and resuspended in RPMI-1640 complete medium. When differentiation into DC cells occurred, 50 ng / ml of IL-4 and 100 ng / ml of GM-CSF were added to the medium, and the cells were cultured at a density of 1×10 6 cells / mL in a 100-mm diameter culture dish and differentiated in an incubator at 37°C with a carbon dioxide concentration of 5%. Day 3: Half of the amount of RPMI complete medium was replaced with fresh RPMI complete medium, and the concentrations of IL-4 and GM-CSF were maintained at 50 ng / ml and 100 ng / ml, respectively. Day 6: Non-adherent cells (DC cells) in the culture dish were collected and washed with PBS. The washed DC cells were resuspended in RPMI complete medium at a density of 1×10 6 cells / mL. Next, 1×10 5 DC cells were added to each well of a 96-well cell culture plate and pre-incubated for 1 hour with different dilutions of the compound to be tested (or DMSO blank negative control at an equivalent concentration), followed by addition of 200 μg / ml of the TLR2 agonist Zymosan to stimulate the DC cells for 24 hours. Day 7: After the DC cells were stimulated with Zymosan for 24 hours, the supernatant of each well of the 96-well plate was collected, and the concentration of IL-23 was detected by ELISA. 【0108】 By the said test, the inhibition of the secretion of IL-23 in DC cells differentiated from human monocytes by the compound of the present disclosure was determined, and the measured IC 50 values are shown in Table 8. 【0109】 【Table 9】 【0110】 Test Example 4. Imiquimod-induced Psoriasis Inhibition Assay An appropriate amount of Compound I was incorporated into an ointment containing the following components: 0.1% of Compound I, 9% of hexanediol, 78.8% of white petrolatum, 5% of paraffin, 7% of glyceryl monostearate and glyceryl distearate, and 0.1% of dihydroxybutyltoluene, and mechanically stirred until the ointment was formed. 【0111】 1) Establishment of Model and Drug Administration Seven-week-old female Balb / c mice were used, and the hair on the back in the range of 2 cm × 3 cm was shaved on the day before the experiment. From the first day to the seventh day of the experiment, the test compound was applied to the skin, and 6 hours later, imiquimod (IMQ) ointment (Aldara (5%)) was continuously applied to the skin on the back of the mice for 7 days to establish a mouse model of psoriasis, and the same amount of petrolatum ointment was administered to the control group. On the 3rd, 5th, and 7th days, the severity of dermatitis was evaluated, the skin thickness, crust, and erythema were measured, and scored on a 5-point scale (0 - 5) respectively. The total score was used to evaluate the severity of dermatitis. On the 7th day of the experiment, the spleen was collected and weighed, and the ratio of the spleen to the body weight was calculated to evaluate the immunosuppressive effect of the drug. 【0112】 In this experiment, a normal control group, a model control group, a low-dose group, a medium-dose group, a high-dose group (0.01%, 0.03%, 0.1%) of Compound I, and a 0.03% reference compound A group were set up. 【0113】 2) Evaluation Index The total score of the degree of skin inflammation is the clinical score, and its index is relatively subjective. The higher the score, the more severe the disease. The degree of increase in skin thickness is an objective evaluation index. The greater the increase in thickness, the more severe the disease. The spleen-to-body weight ratio is an objective evaluation index. The smaller the spleen-to-body weight ratio, the stronger the immunosuppressive effect of the drug. 【0114】 3) Experimental Results 3.1) Clinical score Compound A and Compound I at each volume significantly reduced the clinical score at the end of the experiment (Figs. 2 - 5). As can be seen from the scores of each item, each dose group of Compound I mainly improved the scales in the psoriasis model and reduced the skin thickness. In terms of the skin thickness score, both the low - dose group and the high - dose group of Compound I significantly inhibited the increase in skin thickness, while Compound A had no significant effect. 0.03% of Compound I had a significantly higher inhibitory effect on the increase in skin thickness than Compound A at the same dose (Fig. 5). The results indicate that Compound I is significantly superior to the reference compound A in improving the symptoms of psoriasis. The specific score data are shown in Table 9 below. 【0115】 【Table 10】 【0116】 3.2) Spleen - to - body weight ratio Under the influence of IMQ, there was no significant difference in the body weight of animals between the model group and each administration group, but the spleen - to - total body weight ratio of the model group was significantly increased (Fig. 6), indicating splenomegaly. Compound A had no significant effect on the spleen - to - body weight ratio, while the three administration groups of Compound I significantly decreased the spleen - to - body weight ratio, suggesting that these compounds have an immunosuppressive effect and Compound I shows a dose - response effect. At the same dose (0.03%), the effect of Compound I on the spleen - to - body weight ratio was significantly different from that of Compound A, and these results indicate that the effect of Compound I on immunity is stronger than that of Compound A. The specific data are shown in Table 10 below. 【0117】 【Table 11】

Claims

[Claim 1] The X-ray powder diffraction spectrum, expressed as a diffraction angle of 2θ, has characteristic peaks at 7.537°, 15.219°, 21.798°, and 22.911°, preferably at 7.537°, 11.366°, 14.297°, 15.219°, 16.190°, 17.623°, 19.134°, 21.798°, 22.911°, and 24.735°, more preferably at 7.537°, 11.366°, 14.018°, 14.297°, 15.219°, 16.190°, 17.623°, 19.134°, 21.062°, 21.798°, 22.911°, and 24.735°. , having characteristic peaks at 25.123° and 26.884°, more preferably having characteristic peaks at 7.537°, 11.366°, 14.018°, 14.297°, 15.219°, 16.190°, 17.623°, 18.412°, 19.134°, 20.010°, 21.062°, 21.798°, 22.911°, 23.889°, 24.735°, 25.123°, 26.884°, 28.343°, 29.645°, 33.499° and 36.362°, and even more preferably having an X-ray powder diffraction spectrum represented by a diffraction angle of 2θ as shown in Figure 7, formula I: 【Chemistry 1】 The crystalline form A of the compound. [Claim 2] Crystal form A according to claim 1, wherein the error range of the 2θ angle is ±0.2°. [Claim 3] A method for producing crystal form A according to claim 1, which is method I or method II: Method I is, (a1) Equation I: 【Chemistry 2】 I A step of mixing the compound with solvent (1); (b1) A step of crystallization under stirring; Includes, The solvent (1) is one or more selected from the group consisting of water, nitromethane, N,N-dimethylformamide, acetonitrile, methanol, ethanol, acetone, isopropanol, methyl ethyl ketone, tetrahydrofuran, 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether, 1,2-xylene, toluene, dioxane, and hexane. Method II is, (a2) A step of mixing the compound of formula I with solvent (2); (b2) A step of crystallization under volatilization; Includes, The solvent (2) is one or more selected from the group consisting of acetone, acetonitrile, dichloromethane, dioxane, ethanol, ethyl acetate, heptane, isopropanol, methanol, methyl ethyl ketone, methyl tert-butyl ether, tetrahydrofuran, toluene, and water. [Claim 4] The method according to claim 3, wherein the solvent (1) is water, nitromethane, N,N-dimethylformamide, acetonitrile, ethanol, acetone, isopropanol, methyl ethyl ketone, tetrahydrofuran, 1,2-dimethoxyethane, chloroform, methyl tert-butyl ether, 1,2-xylene, toluene, dioxane, hexane, or a mixed solvent of water and methanol. [Claim 5] The method according to claim 4, wherein solvent (1) is a mixed solvent of water and methanol, and the molar ratio of water to methanol is 0.37 to 0.90, preferably 0.37, 0.40, 0.47, 0.50, 0.57, 0.60, 0.66, 0.70, 0.74, 0.80, 0.82, or 0.

90. [Claim 6] Solvent (2) is acetone, acetonitrile, dichloromethane, dioxane, ethyl acetate, heptane, isopropanol, methyl ethyl ketone, methyl tert-butyl ether, toluene, water, or a mixture of the following: acetone and acetonitrile, acetone and dichloromethane, acetone and dioxane, acetone and ethanol, acetone and ethyl acetate, acetone and heptane, acetone and isopropanol, acetone and methanol, acetone and methyl ethyl ketone, acetone and methyl tert-butyl ether, acetone and tetrahydrofuran, acetone and toluene, acetone and water, acetonitrile and dichloromethane, acetonitrile and dioxane, acetonitrile and ethanol, acetonitrile and ethyl acetate, acetonitrile and isopropanol, acetonitrile and methyl ethyl ketone, acetonitrile and methyl tert-butyl ether, acetonitrile and tetrahydrofuran, acetonitrile and toluene, dichloromethane and dioxane, dichloromethane and ethanol, dichloromethane and ethyl acetate, dichloromethane and heptane, dichloromethane and isopropanol, dichloromethane and methyl ethyl ether Dichloromethane and methyl tert-butyl ether, dichloromethane and tetrahydrofuran, dichloromethane and toluene, dioxane and methyl ethyl ketone, dioxane and tetrahydrofuran, dioxane and toluene, ethanol and ethyl acetate, ethanol and methanol, ethanol and tetrahydrofuran, ethanol and toluene, ethanol and water, ethyl acetate and heptane, ethyl acetate and isopropanol, ethyl acetate and methanol, ethyl acetate and methyl ethyl ketone, ethyl acetate and methyl tert-butyl ether, ethyl acetate and toluene, heptane and methyl ethyl ketone, heptane and methyl tert-butyl ether, heptane and tetrahydrofuran, heptane and toluene, methanol and methyl ethyl ketone, methanol and methyl tert-butyl ether, methanol and toluene, methanol and water, methyl ethyl ketone and methyl tert-butyl ether, methyl ethyl ketone and tetrahydrofuran, methyl tert-butyl ether and toluene, tetrahydrofuran and toluene,The method according to claim 3, wherein the solvent (2) is either tetrahydrofuran or water, and preferably, when the solvent (2) is a mixed solvent, the volume ratio of the two is 1:3 to 3:1, preferably 1:

1. [Claim 7] Crystal form A or according to claim 1 or 2 A pharmaceutical composition comprising crystalline form A produced by the method of any one of claims 3 to 6, and optionally a pharmaceutically acceptable excipient. [Claim 8] Crystal form A or according to claim 1 or 2 A method for producing a pharmaceutical composition, comprising the step of mixing crystalline form A, produced by the method of any one of claims 3 to 6, with a pharmaceutically acceptable excipient. [Claim 9] Use of crystalline form A as described in claim 1 or 2, or crystalline form A produced by the method of any one of claims 3 to 6, in the manufacture of a pharmaceutical product for the prevention and / or treatment of PDE-related diseases, preferably asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes mellitus, allergic rhinitis, allergic conjunctivitis, ulcerative colitis, or rheumatoid arthritis. [Claim 10] Use of the pharmaceutical composition according to claim 7 in the manufacture of a pharmaceutical for the prevention and / or treatment of a PDE-related disease, preferably asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes mellitus, allergic rhinitis, allergic conjunctivitis, ulcerative colitis, or rheumatism. [Claim 11] Use of crystal form A as described in claim 1 or 2, or crystal form A produced by the method of any one of claims 3 to 6, in the manufacture of a pharmaceutical product for the prevention and / or treatment of asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes mellitus, allergic rhinitis, allergic conjunctivitis, ulcerative colitis, or rheumatism. [Claim 12] Use of the pharmaceutical composition according to claim 7 in the manufacture of a pharmaceutical for the prevention and / or treatment of asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes mellitus, allergic rhinitis, allergic conjunctivitis, ulcerative colitis, or rheumatism.

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