Crystal Form and Preparation Method of Triazinedione Derivatives

Abstract

The present disclosure relates to crystal forms of triazinedione derivatives and methods for their preparation. Specifically, the present disclosure relates to different crystal forms of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione and methods for their preparation. The crystal forms of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione provided by the present disclosure have good stability and can be better used in clinical treatment.

Description

Technical Field

[0001] The present disclosure belongs to the pharmaceutical field and relates to crystal forms of triazinedione derivatives. Specifically, it relates to crystal forms of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione and a method for preparing the same.

Background Art

[0002] Hypertrophic cardiomyopathy (HCM) is an autosomal dominant genetic myocardial disease associated with gene mutations. The worldwide incidence is approximately 0.2%, and it is the most important cause of sudden death in young people under 35 years old (Tuohy, C.V. et al., Eur J Heart Fail, 22, 2020, 228-240). The clinical findings are characterized by asymmetric hypertrophy of the left ventricular wall, which always invades the ventricular septum, resulting in a smaller ventricular cavity, obstruction of left ventricular blood filling, and decreased ventricular diastolic compliance. Depending on the presence or absence of obstruction in the left ventricular outflow tract, it is divided into obstructive and non-obstructive hypertrophic cardiomyopathy. Currently, clinically, the treatment of hypertrophic cardiomyopathy often uses β-blockers and calcium channel blockers to reduce cardiac contraction and relieve symptoms. However, these treatments only attempt to address the surface symptoms and do not fundamentally treat the lesions that cause them. As HCM progresses, heart transplantation is the only option (Ramaraj, R. Cardiol Rev, 16, 2008, 172-180). Therefore, finding a treatment method for the root cause of HCM onset is extremely urgent.

[0003] In previous studies, it was found that 70% of HCM patients were all due to myotilin gene mutations. Among these patients, multiple-site mutations were found in 5% - 7% of them. Currently, more than about 70 pathogenic mutations have been identified, but most of these mutations are family-specific, and only a few hotspots have been confirmed, such as the MYH7 R403Q and R453C mutations (Frey, N. et al., Nat Rev Cardiol, 9, 2011, 91 - 100, Sabater-Molina, M. et al., Clin Genet, 93, 2018, 3 - 14). Studies on the incidence probability caused by gene mutations have found that patients with MYH7 gene mutations account for about 30%. Compared with other myotilin genes, MYH7 causes earlier onset of the disease and more severe myocardial hypertrophy. Myosin is a constituent unit of the thick filaments of myofibrils and plays an important role in muscle movement. Its molecular shape is like vermicelli and is composed of two heavy chains and multiple light chains. The head of myosin binds to actin to form cross-bridges, which greatly improves the ATPase activity of myosin, catalyzes the ATP hydrolysis reaction, generates energy to promote the sliding of cross-bridges, and causes muscle contraction. The research results show that MYH7 gene mutations can cause an increase in myosin ATPase activity, a decrease in the proportion of myosin super-relaxed state (SRX), and an increase in cross-bridges between myosin and actin, thereby causing abnormal cardiac contraction function (Green, EM. et al., Science, 351, 2016, 617 - 621, Sommese, RF. et al., Proc Natl Acad Sci U S A, 110, 2013, 12607 - 12612). Therefore, myosin is an important target for treating hypertrophic cardiomyopathy.

[0004] In the application with the application number WO2022105852, a series of triazinedione derivatives containing (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione are provided and structurally characterized. Further, in this application, a biological evaluation was carried out on (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the results show that the compound has excellent inhibitory effects on the myosin ATPase enzyme.

[0005] The crystal structure of a pharmaceutically active ingredient tends to affect the chemical stability of the drug, and differences in crystallization conditions and storage conditions may cause changes in the crystal structure of the compound, and sometimes may be accompanied by the formation of other crystal forms. Generally, amorphous pharmaceutical products do not have a regular crystal structure and tend to have other defects such as relatively poor product stability, relatively fine crystallization, relatively difficult filtration, easy caking, and low fluidity. Therefore, it is necessary to improve the properties of each aspect of the above products, and it is necessary to conduct in-depth research to find a crystal form with relatively high crystal purity and good physical and chemical stability.

Summary of the Invention

[0006] The present disclosure provides a crystal form of the myosin inhibitor (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and a preparation method and use thereof.

[0007] The present disclosure provides a crystal form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione.

[0008] In some embodiments, the present disclosure provides crystalline Form A of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the X-ray powder diffraction pattern represented by diffraction angle 2θ has peaks characteristic of 9.2, 10.9, 18.6, 20.1, 22.0, 26.9, optionally peaks characteristic of 9.2, 10.9, 13.3, 16.9, 18.6, 19.2, 20.1, 22.0, 26.9, 35.6, and optionally peaks characteristic of 9.2, 10.9, 13.3, 16.9, 18.6, 19.2, 20.1, 20.6, 22.0, 26.9, 28.0, 35.6.

[0009] In some embodiments, the present disclosure provides crystalline Form A of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the X-ray powder diffraction pattern represented by diffraction angle 2θ has peaks characteristic of 9.2, 10.9, 18.5, 20.1, 22.0, 26.9, optionally peaks characteristic of 9.2, 10.9, 13.3, 16.8, 18.5, 19.1, 20.1, 22.0, 26.9, 35.5, and optionally peaks characteristic of 9.2, 10.9, 13.3, 16.8, 18.5, 19.1, 20.1, 20.6, 22.0, 26.9, 27.9, 35.5.

[0010] In some embodiments, the present disclosure provides crystalline form B of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the X-ray powder diffraction pattern represented by diffraction angle 2θ has characteristic peaks at 11.1, 15.9, 18.8, 20.4, 22.2, 26.8, optionally at 6.6, 9.5, 11.1, 15.9, 18.8, 20.4, 21.0, 22.2, 26.8, and optionally at 6.6, 9.5, 11.1, 13.3, 14.0, 15.9, 18.8, 19.0, 19.8, 20.4, 21.0, 22.2, 26.8, 28.0.

[0011] In some embodiments, the present disclosure provides crystalline form C of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the X-ray powder diffraction pattern represented by diffraction angle 2θ has characteristic peaks at 8.3, 14.4, 16.8, 17.8, 19.3, 22.3, 23.1.

[0012] In some embodiments, the present disclosure provides crystalline form D of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the X-ray powder diffraction pattern represented by diffraction angle 2θ has characteristic peaks at 7.8, 8.7, 9.7, 13.5, 14.0, 22.9, optionally at 7.8, 8.7, 9.7, 13.5, 14.0, 16.7, 19.6, 20.2, 22.9, and optionally at 7.8, 8.7, 9.7, 13.5, 14.0, 15.5, 16.7, 17.5, 19.6, 20.2, 22.9, 23.9.

[0013] In some embodiments, the present disclosure provides crystalline form E of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione. The X-ray powder diffraction pattern represented by diffraction angle 2θ has characteristic peaks at 6.9, 9.3, 14.2, 17.1, 18.0, 25.0, and optionally, has characteristic peaks at 6.9, 9.3, 13.5, 14.2, 17.1, 18.0, 22.4, 23.2, 25.0, and optionally, has characteristic peaks at 6.9, 9.3, 13.5, 14.2, 16.1, 17.1, 18.0, 21.1, 22.4, 23.2, 23.8, 25.0.

[0014] In alternative embodiments, the present disclosure provides a crystalline form of the compound represented by formula (1), wherein the error range of the above 2θ angle is ±0.2.

[0015] The present disclosure provides a method for preparing crystalline form A of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, which is any one of the following methods, namely, Method 1: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (1), and stir or heat to dissolve it. Solvent (1) is at least one selected from 7% water / ethanol, tetrahydrofuran, 10% water / isopropanol, 10% water / acetone, methanol, ethanol, 10% water / methanol, (b) Perform crystallization, Or, Method 2: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (2), and stir to dissolve or heat to dissolve. Solvent (2) is selected from 10% water / acetone and dichloromethane. (b) Add solvent (3) and crystallize. Solvent (3) is at least one selected from water, methanol, and ethanol. Or, Method Three: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (4). Solvent (4) is at least one selected from water, methanol, ethanol, isopropanol, acetone, ethyl acetate, acetonitrile, isopropyl acetate, methyl tert-butyl ether, methyl isobutyl ketone, n-heptane, cyclohexane, 1,4-dioxane, 10% water / methanol, and 7% water / ethanol. (b) Select from slurrying and crystallizing.

[0016] The present disclosure provides a method for preparing crystalline form B of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and any one of the following methods, namely, Method One: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (5), and stir to dissolve or heat to dissolve. Solvent (5) is selected from ethanol. (b) Crystallize. Or, Method Two: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (6), and stir to dissolve or heat to dissolve. Solvent (6) is selected from dichloromethane. (b) Add solvent (7) and perform crystallization. Solvent (7) is at least one selected from ethyl acetate, acetone, acetonitrile, and methyl isobutyl ketone.

[0017] The present disclosure provides a method for preparing crystalline form C of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione. The above method includes: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (9), and stir to dissolve or heat to dissolve. Solvent (9) is selected from dichloromethane. (b) Add solvent (10) and perform crystallization. Solvent (10) is selected from isopropyl acetate.

[0018] The present disclosure provides a method for preparing crystalline form D of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione. The above method includes: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with solvent (11). Solvent (11) is selected from dichloromethane. (b) Add solvent (12) to form a slurry and perform crystallization. Solvent (12) is selected from methyl tert-butyl ether.

[0019] The present disclosure provides a method for preparing crystalline form E of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, and the method comprises (a) mixing the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with a solvent (13), wherein the solvent (13) is selected from dichloromethane, (b) adding a solvent (14) to form a slurry, followed by crystallization, wherein the solvent (14) is selected from n-heptane.

[0020] In certain embodiments, the method for preparing the crystalline form described in the present disclosure further comprises one or more of filtration, washing, or drying steps.

[0021] The present disclosure further provides a pharmaceutical composition prepared from the crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione.

[0022] The present disclosure further provides a pharmaceutical composition comprising the crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione or a mixture thereof, or the crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione prepared by the above method, and optionally a pharmaceutically acceptable excipient.

[0023] The present disclosure further provides a method for preparing a pharmaceutical composition, which includes a step of mixing a crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione or a mixture thereof, or a crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione prepared by the above method with a pharmaceutically acceptable excipient.

[0024] The present disclosure further provides the use of a crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione or a mixture thereof, or a crystalline form or a mixture thereof prepared by the above method, or the above composition, or the composition prepared by the above method in the preparation of a medicament for treating a disease or condition associated with myosin regulation.

[0025] The present disclosure further provides the use of a crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione or a mixture thereof, or a crystalline form prepared by the above method or a mixture thereof, or the above composition, or a composition prepared by the above method, in the preparation of a medicament for treating a disease or medical condition, wherein the disease or medical condition is selected from heart failure with preserved ejection fraction, ischemic heart disease, angina pectoris, restrictive cardiomyopathy, diastolic dysfunction, hypertrophic cardiomyopathy (HCM), non-obstructive hypertrophic cardiomyopathy (nHCM), obstructive hypertrophic cardiomyopathy (oHCM), valvular disease, heart failure with preserved ejection fraction (HFpEF), heart failure with mid-range ejection fraction (HFmREF), aortic stenosis, inflammatory cardiomyopathy, rheumatic endocarditis, endomyocardial fibrosis, infiltrative cardiomyopathy, hemochromatosis, Fabry disease, glycogenosis, congenital heart disease, tetralogy of Fallot, left ventricular hypertrophy, refractory angina pectoris, and Chagas disease; alternatively, selected from ischemic heart disease, restrictive cardiomyopathy, hypertrophic cardiomyopathy (HCM), non-obstructive hypertrophic cardiomyopathy (nHCM), obstructive hypertrophic cardiomyopathy (oHCM), inflammatory cardiomyopathy, infiltrative cardiomyopathy, congenital heart disease, and left ventricular hypertrophy; alternatively, the disease or medical condition is hypertrophic cardiomyopathy (HCM).

[0026] As used herein, the "2θ or 2θ angle" refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degrees. The error range of each characteristic peak 2θ is ±0.20 (including the case where numbers exceeding one decimal place are rounded), specifically, -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, 0.20.

[0027] As used herein, "crystallization" or "crystallization" includes, but is not limited to, stirred crystallization, slurry crystallization, cooling crystallization, and volatile crystallization.

[0028] As used herein, "differential scanning calorimetry or DSC" refers to measuring the temperature difference and heat flow difference between a sample and a reference to characterize all physical and chemical changes related to heat effects during heating or constant temperature of the sample and obtain information on the phase transition of the sample.

[0029] As used herein, "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, vector, solubilizer, sweetener, diluent, preservative, dye / colorant, flavorant, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, or emulsifier that has already been approved by the US Food and Drug Administration (FDA) and is acceptable for use in humans or livestock animals.

Brief Description of the Drawings

[0030]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

MODE FOR CARRYING OUT THE INVENTION

[0031] The present disclosure will be described in more detail by the following examples and experimental examples. These examples and experimental examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0032] Test conditions of the equipment used in the experiment: The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is in units of 10 -6 (ppm). The NMR measurement is performed using a nuclear magnetic resonance apparatus Bruker AVANCE-400, and the measurement solvents are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD 3 OD), and the internal standard is tetramethylsilane (TMS).

[0033] For MS measurement, liquid chromatograph mass spectrometers Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS (manufacturer: Agilent, MS model number: 6110 / 6120 Quadrupole MS), waters ACQuity UPLC-QD / SQD (manufacturer: waters, MS model number: waters ACQuity Qda Detector / waters SQ Detector), and THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model number: THERMO Q Exactive) were used.

[0034] For high performance liquid chromatography (HPLC) analysis, high performance liquid chromatographs Agilent HPLC 1200DAD, Agilent HPLC 1200VWD, and Waters HPLC e2695-2489 were used.

[0035] For chiral HPLC analysis measurement, high performance liquid chromatograph Agilent 1260 DAD was used.

[0036] For preparative high performance liquid chromatography, preparative chromatographs Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 were used.

[0037] For chiral preparative, preparative chromatograph Shimadzu LC-20AP was used.

[0038] As the CombiFlash high-speed preparative chromatograph, Combiflash Rf200 (TELEDYNE ISCO) was used.

[0039] As the silica gel plate for thin-layer chromatography, Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate is used. The specification of the silica gel plate used in thin-layer chromatography (TLC) is 0.15 mm to 0.2 mm, and the specification for the separation and purification of products by thin-layer chromatography is 0.4 mm to 0.5 mm.

[0040] For silica gel column chromatography, generally, 200 - 300 mesh silica gel made by Yantai Huanghai was used as the vector.

[0041] The known starting materials according to the present disclosure can be synthesized by or in accordance with known methods in this field, or can be purchased from companies such as ABCR GmbH&Co.KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0042] In the examples, unless otherwise specified, all reactions can be carried out under an argon or nitrogen atmosphere.

[0043] An argon or nitrogen atmosphere means that an argon or nitrogen balloon with a volume of about 1 L is connected to the reaction flask.

[0044] A hydrogen atmosphere means that a hydrogen balloon with a volume of about 1 L is connected to the reaction flask.

[0045] For the hydrogenation reaction under pressure, a Parr 3916EKX type hydrogenation apparatus and a Qinglan QL - 500 type hydrogen generator or an HC2 - SS type hydrogenation apparatus were used.

[0046] The hydrogenation reaction generally involved repeating the operation of evacuating and filling with hydrogen three times.

[0047] For the microwave reaction, a CEM Discover - S 908860 type microwave reactor was used.

[0048] In the examples, unless otherwise specified, the solution refers to an aqueous solution.

[0049] In the examples, unless otherwise specified, the reaction temperature is room temperature of 20 °C to 30 °C.

[0050] In monitoring the progress of the reaction in the examples, thin layer chromatography (TLC) is used. The developing solvent used in the reaction, the eluent system for column chromatography for purifying the compound, and the developing solvent system for thin layer chromatography include A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvents is adjusted according to the polarity of the compound and can be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

[0051] When the compounds in the examples contain two or more chiral centers, the relative stereochemistry of these compounds is identified by NMR studies and / or X-ray diffraction. In these cases, the prefix "rel" is used, and then these compounds are identified by the R / S nomenclature. At this time, R / S only provides relative stereochemical information without indicating absolute stereochemistry.

[0052] XRPD is X-ray powder diffraction detection: measurements are performed using a BRUKER D8 type X-ray diffractometer. Specific collection information: Cu anode (40 kV, 40 mA), radiation: monochromatic Cu-Kα ray (λ = 1.5418 Å). Scanning mode: θ / 2θ, scanning range (2θ range): 5° to 50° (or 3° to 48°).

[0053] DSC is differential scanning calorimetry: measurements are performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter. The heating rate is 10 °C / min. The specific temperature range refers to the corresponding spectrum (mainly 25 °C to 300 °C), and the purge rate of nitrogen gas is 50 mL / min.

[0054] TGA is thermogravimetric analysis: Detection was performed using a METTLER TOLEDO TGA 2 type thermogravimetric analyzer. The heating rate was 10 °C / min, and the specific temperature range was referred to the corresponding spectrum (mainly 25 °C to 300 °C), and the purge rate of nitrogen gas was 50 mL / min.

[0055] DVS is dynamic vapor sorption: Detection was performed using an SMS DVS Advantage. At 25 °C, the humidity change was 50% - 95% - 0% - 95% - 50%, with a step of 10% (the last step was 5%) (the specific humidity range was based on the corresponding pattern, which is mostly the usage method described here), and the judgment criterion was dm / dt ≤ 0.002%.

[0056] Example 1. (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione Preparation (Refer to the preparation method of Example 16 in the application with application number WO2022105852)

Chemical formula

[0057] Step 1 4-Isocyanatotetrahydro-2H-pyran 1b At 15 °C, a solution of tetrahydro-2H-pyran-4-amine 1a (10.0 g, 100.0 mmol, Shanghai Shaoyuan Technology Co., Ltd.) and N,N-diisopropylethylamine (28.4 g, 220.0 mmol, Shanghai Titan Technology Co., Ltd.) in anhydrous dichloromethane (120 mL) was slowly added dropwise to a solution of bis(trichloromethyl) carbonate (11.9 g, 40.0 mmol, Shanghai Titan Technology Co., Ltd.) in anhydrous dichloromethane (30 mL). The reaction was carried out at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product of the title product 1b. The crude product was used directly in the next step without purification.

[0058] Step 2 6-(1H-Pyrazol-1-yl)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione 1d At -10 °C, 1,8-diazabicyclo[5.4.0]undec-7-ene (15.2 g, 100.0 mmol, Shanghai Shaoyuan Technology Co., Ltd.) was slowly added dropwise to a solution of compound 1c (10.5 g, 95.2 mmol) and the crude product of compound 1b in anhydrous N,N-dimethylacetamide (120 mL). The reaction was stirred at 0 °C for 1 hour. Then, at 0 °C, carbonyldiimidazole (23.2 g, 142.8 mmol, Shanghai Bide Technology Co., Ltd.) and 1,8-diazabicyclo[5.4.0]undec-7-ene (21.7 g, 142.8 mmol, Shanghai Shaoyuan Technology Co., Ltd.) were added. The reaction was stirred at 60 °C for 16 hours. The reaction solution was concentrated under reduced pressure. Dichloromethane was added to the obtained residue, stirred, filtered, the filter cake was collected, and dried under reduced pressure to obtain the title product 1d (16.6 g, two-step yield: 63.4%). MS m / z (ESI): 264.1 [M+1].

[0059] Step 3 (R)-N-((S)-1-(2-Fluoro-5-methylphenyl)ethyl)-2-methylpropane-2-sulfenamide 1f (R)-N-(2-Fluoro-5-methylphenylmethylene)-2-methylpropan-2-sulfenamide 1e (6.90 g, 28.59 mmol, prepared by the method disclosed in Intermediate 3B on page 56 of the specification of patent application "WO2020092208A1") was dissolved in dichloromethane (100 mL) and replaced with nitrogen gas three times. The reaction was cooled to -60 °C, and a 2-methyltetrahydrofuran solution of 3 M methylmagnesium bromide (19.1 mL, 57.18 mmol) was added dropwise. Under nitrogen protection, the reaction was stirred at room temperature for 2 hours. Saturated ammonium chloride solution (100 mL) was added, and the organic phase was separated. The aqueous phase was extracted with ethyl acetate (80 mL × 2), the organic phases were combined, washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by high-performance liquid chromatography (Boston Phlex C18 150 mm * 30 mm, 5 μm, eluent: water (10 mmol ammonium bicarbonate), acetonitrile, 20% - 95% acetonitrile, gradient elution for 20 minutes, flow rate: 30 mL / min) to obtain the title product 1f (5.60 g, yield: 76.1%). MS m / z (ESI): 258.0 [M + 1].

[0060] Step 4 (S)-1-(2-Fluoro-5-methylphenyl)ethylamine hydrochloride 1g Compound 1f (670 mg, 2.60 mmol) was dissolved in ethanol (10 mL), cooled to 0 °C, and thionyl chloride (620 mg, 5.21 mmol) was added dropwise. The reaction was stirred at room temperature for 1 hour. It was concentrated under reduced pressure to obtain the crude product of the title product 1g (493 mg, yield: 99.9%). The crude product was used directly in the next step without purification. MS m / z (ESI): 153.9 [M + 1].

[0061] Step 5 (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione 1 Compound 1d (425 mg, 1.61 mmol) and compound 1g (297 mg, 1.94 mmol) were dissolved in 1,4 - dioxane (10 mL), and the reaction was stirred at 120 °C for 16 h. The reaction solution was concentrated under reduced pressure and purified by high - performance liquid chromatography (Boston Phlex C18 150 mm * 30 mm, 5 μm, eluent: water (10 mmol ammonium bicarbonate), acetonitrile, 20% - 95% acetonitrile, gradient elution for 20 min, flow rate: 30 mL / min), and the title product 1 (310 mg, yield: 55.1%) was obtained. MS m / z (ESI): 349.0 [M + 1].

[0062] 1 H NMR (500 MHz, DMSO - d 6 ) δ 10.50 (brs, 1H), 7.28 - 7.05 (m, 4H), 5.22 (m, 1H), 4.64 (m, 1H), 3.90 - 3.86 (m, 2H), 3.33 - 3.27 (m, 2H), 2.54 - 2.42 (m, 2H), 2.28 (s, 3H), 1.45 - 1.39 (m, 5H).

[0063] Example 2. Preparation of amorphous (S) - 6 - ((1 - (2 - fluoro - 5 - methylphenyl) ethyl) amino) - 3 - (tetrahydro - 2H - pyran - 4 - yl) - 1,3,5 - triazine - 2,4(1H,3H) - dione The substance obtained by purification by high - performance liquid chromatography in Example 1 was lyophilized to obtain a product. Detected by X - ray powder diffraction, the product was defined as an amorphous form of (S) - 6 - ((1 - (2 - fluoro - 5 - methylphenyl) ethyl) amino) - 3 - (tetrahydro - 2H - pyran - 4 - yl) - 1,3,5 - triazine - 2,4(1H,3H) - dione. In the X - ray powder diffraction pattern represented by the diffraction angle 2θ, it has no characteristic peaks in the range of 3° - 48°, and the XRD pattern is as shown in Figure 1.

[0064] Example 3. Inhibitory effect of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione on myosin ATPase activity The following method is for measuring the inhibitory effect of a compound on myosin ATPase activity, and the experimental method will be briefly described as follows.

[0065] 1. Experimental materials and equipment a. Cardiac actin (Cytoskeleton, AD99) b. Myosin motor protein S1 fragment (Cytoskeleton, CS-MYS03) c. ATP (Sigma, A7699-1G) d. UltraPureTM 1M Tris-HCl buffer, pH 7.5 (Thermo, 15567027) e. CytoPhosTM phosphate detection bio-reagent kit (Cytoskeleton, BK054) f. Magnesium chloride solution (Sigma, 68475-100ML-F) g. Potassium chloride solution (Sigma, 60142-100ML-F) h. EGTA (Sigma, E3889-100G) i. 96-well plate (Corning, 3697) j. U-bottom 96-well plate (Corning, 3795) k. Plate reader (BMG, PHERAstar) l. Constant temperature incubator (Shanghai Boxun, SPX-100B-Z) 2. Experimental steps 1.61 μM of cardiac actin and 0.07 μM of myosin motor protein S1 fragment were mixed with different concentrations of small molecule compounds (initial concentration 100 μM, diluted in a 3-fold gradient to 9 concentrations), and incubated at 37 °C for 1 hour. Then, 120 μM of ATP was added and incubated at 37 °C for 2 hours. Finally, the detection solution (70 μL / well) in the CytoPhosTM Phosphate Detection Bio Reagent Kit was added to each well and incubated at room temperature for 10 min. The OD value at a wavelength of 650 nM was read using a plate reader, the amount of Pi was calculated according to the standard curve, the data was processed with GraphPad software, an inhibition curve was drawn based on each concentration of the compound and the corresponding inhibition rate, and the IC 50 value was calculated, which is the concentration of the compound when the inhibition rate reaches 50%.

[0066] Conclusion: (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione has an IC 50 of 1.06 μM for the inhibitory activity against myosin ATPase and has excellent inhibitory effects on myosin ATPase.

[0067] Example 4. Toxicokinetic Evaluation of Repeated Intragastric Administration of (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione to SD Rats for 14 Days 1. Summary Using SD rats as test animals and the LC / MS / MS method, the concentrations of the drug prototype in plasma and administration solution at different time points of SD rats administered with the compound to be measured intragastrically were measured. The toxicokinetic behavior of (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione in the body of SD rats was studied and its toxicokinetic characteristics were evaluated.

[0068] 2. Experimental Plan 2.1 Test drug (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, compound MYK-461( [Chemical formula] , Example 1 of WO2014205223A1).

[0069] 2.2 Experimental animals 24 male and female Sprague-Dawley rats provided by Vital River Laboratory Animal Technology Co., Ltd., equally divided into 6 groups, with 4 rats in each group.

[0070] 2.3 Preparation of drugs Weighed a certain amount of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 15% PEG400 and 85% (10% TPGS + 1% HPMC K100LV) to prepare a light yellow homogeneous suspension.

[0071] Weighed a certain amount of compound MYK-461, added 0.5% MC to prepare a colorless transparent solution.

[0072] 2.4 Administration Administered by gavage. The administration doses of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione were 5 mg / kg, 15 mg / kg, and 30 mg / kg respectively, and the administration volumes were all 10 mL / kg. The administration doses of compound MYK-461 were 0.5 mg / kg, 1.5 mg / kg, and 3 mg / kg respectively, and the administration volumes were all 10 mL / kg.

[0073] 3. Procedure On the first day, 0.2 mL of blood was collected from the orbital cavity at 0.5 h, 1.0 h, 2.0 h, 4.0 h, 8.0 h, and 24.0 h after administration. On the 7th and 14th days, 0.2 mL of blood was collected from the orbital cavity at 0.5 h, 1.0 h, 2.0 h, 4.0 h, 8.0 h, and 24.0 h before and after administration. The blood was placed in an EDTA-K2 anticoagulant test tube and centrifuged at 10,000 rpm for 1 min (4 °C). Plasma was separated within 1 h and stored at -20 °C for measurement. The process from blood collection to centrifugation was operated under ice bath conditions. Food was given 2 h after administration.

[0074] The content of the compound to be measured in the plasma of SD rats after intragastric administration of different concentrations of the drug was measured. That is, 20 μL of plasma from SD rats at each time point after administration was taken, 50 μL of internal standard solution ((S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione internal standard: verapamil 100 ng / mL, compound MYK-461 internal standard: camptothecin 100 ng / mL), and 200 μL of acetonitrile were added. It was vortexed for 5 min and centrifuged for 10 min (3700 revolutions / min). 1 μL of the supernatant was collected from the plasma sample for LC / MS / MS analysis.

[0075] 4. Results of pharmacokinetic parameters

Table 1-1

Table 1-2

[0076] Conclusion: After repeated intragastric administration to SD rats for 14 days, the accumulation of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione in the SD rat body was not significant, while the accumulation of compound MYK-461 in the SD rat body was serious, increasing the administration risk. Obviously, (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione has obvious pharmacokinetic advantages over compound MYK-461.

[0077] Example 5. Preparation of Crystal Form A of the Compound Weighed 300 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 6 mL of 7% water / ethanol, heated to clarify at 80 °C, cooled to room temperature, stirred overnight, filtered, and then dried the solid under vacuum to obtain the product. Detected by X-ray powder diffraction, the product was defined as crystal form A, the X-ray powder diffraction data was shown as in Table 2, and the X-ray powder diffraction pattern was shown as in Figure 2.

[0078] The DSC pattern showed that the peak value of the endothermic peak was 256.71 °C. The TGA pattern showed a weight loss of 0.59% at 30 °C - 100 °C.

[0079] DVS detection showed that under normal storage conditions (i.e., at room temperature and 60% RH), the moisture absorption weight increase of the sample was about 0.12%, under accelerated experiment conditions (i.e., 70% RH), the moisture absorption weight increase was about 0.15%, and under extreme conditions (i.e., 90% RH), the moisture absorption weight increase was about 0.24%. And the crystal form did not transform after redetection after DVS detection.

[0080]

Table 2-1

Table 2-2

[0081] Example 6. Preparation of Crystal Form A of the Compound (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was added to a solvent to form a slurry. After centrifugation, the solid was dried under vacuum to obtain the product. The crystal form was detected and confirmed by X-ray powder diffraction as shown in Table 3.

[0082]

Table 3-1

Table 3-2

Table 3-3

[0083] Example 7. Preparation of Crystal Form A of the Compound (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was added to a solvent to form a slurry. After centrifugation, the solid was dried under vacuum to obtain the product. The crystal form was detected and confirmed by X-ray powder diffraction as shown in Table 4.

[0084]

Table 4-1

Table 4-2

[0085] Example 8. Preparation of Crystal Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 0.1 mL of tetrahydrofuran was added, and the mixture was stirred until clear. The product was obtained by volatile crystallization. The product was crystalline form A detected by X-ray powder diffraction.

[0086] Example 9. Preparation of Crystalline Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of 10% water / isopropanol was added, and the mixture was stirred until clear. The product was obtained by volatile crystallization. The product was crystalline form A detected by X-ray powder diffraction.

[0087] Example 10. Preparation of Crystalline Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of 10% water / acetone was added, and the mixture was stirred until clear. The product was obtained by volatile crystallization. The product was crystalline form A detected by X-ray powder diffraction.

[0088] Example 11. Preparation of Crystalline Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of methanol was added, and the mixture was stirred at 50 °C until clear. The product was obtained by cooling crystallization. The product was crystalline form A detected by X-ray powder diffraction.

[0089] Example 12. Preparation of Crystalline Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of ethanol was added, and the mixture was stirred at 50 °C until clarified. The product was obtained by cooling crystallization. The product was crystal form A detected by X-ray powder diffraction.

[0090] Example 13. Preparation of Crystal Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of 10% water / methanol was added, and the mixture was stirred at 50 °C until clarified. The product was obtained by cooling crystallization. The product was crystal form A detected by X-ray powder diffraction.

[0091] Example 14. Preparation of Crystal Form A of the Compound 10 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 1 mL of 7% water / ethanol was added, and the mixture was stirred at 50 °C until clarified. The product was obtained by cooling crystallization. The product was crystal form A detected by X-ray powder diffraction.

[0092] Example 15. Preparation of Crystal Form A of the Compound 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 0.05 mL of dichloromethane was added, and the mixture was stirred at room temperature until clarified. 0.35 mL of methanol was added and stirred, and the mixture was slurried at room temperature for 3 days. After centrifugation, the solid was dried in vacuo to obtain the product. The product was crystal form A detected by X-ray powder diffraction.

[0093] Example 16. Preparation of Crystal Form A of the Compound Weigh 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, add 0.05 mL of dichloromethane, stir at room temperature until clarified, add 0.35 mL of ethanol, stir, slurry at room temperature for 3 days, after centrifugation, dry the solid under vacuum to obtain the product. The product was crystalline form A detected by X-ray powder diffraction.

[0094] Example 17. Preparation of crystalline form A of the compound Weigh 1 g of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, add 30 mL of 10% water / acetone, stir at 50 °C until clarified, add 30 mL of water, stir, stir at 50 °C for 5 hours, lower the temperature to 25 °C and stir overnight, after filtration, dry the solid under vacuum to obtain the product. The product was crystalline form A detected by X-ray powder diffraction.

[0095] Example 18. Preparation of crystalline form B of the compound Dissolve 5 g of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione in 250 mL of absolute ethanol, heat up to reflux until the solid is clarified. Stop heating, let stand, filter to collect the filter cake, and dry to obtain the product. Detected by X-ray powder diffraction, the product was defined as crystalline form B, the X-ray powder diffraction data was shown as in Table 5, and the X-ray powder diffraction pattern was shown in Figure 3.

[0096] The DSC pattern showed that the peak value of the endothermic peak was 256.45 °C. The TGA pattern showed that the compound had a weight loss of 1.32% between 40 °C and 190 °C. The DVS experimental data showed that under normal storage conditions (i.e., at 25 °C and 60% RH), the weight increase due to moisture absorption of the sample was approximately 0.242%, under accelerated test conditions (i.e., 70% RH), the weight increase due to moisture absorption was approximately 0.298%, and under extreme conditions (90% RH), the weight increase due to moisture absorption was approximately 0.418%. The XRD pattern showed that the crystal form did not change before and after DVS detection.

[0097]

Table 5-1

Table 5-2

[0098] Example 19. Preparation of Crystal Form B of the Compound 30 mg of (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 0.3 mL of dichloromethane was added, and the mixture was stirred at room temperature until clarified. 2.1 mL of ethyl acetate was added and stirred, and then stirred at room temperature for 2 days. After filtration, the solid was dried in vacuo to obtain the product. The product was Crystal Form B detected by X-ray powder diffraction.

[0099] Example 20. Preparation of Crystal Form B of the Compound 5 mg of (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 0.05 mL of dichloromethane was added, and the mixture was stirred at room temperature until clarified. 0.35 mL of acetone was added and stirred, and then slurried at room temperature for 3 days. After centrifugation, the solid was dried in vacuo to obtain the product. The product was Crystal Form B detected by X-ray powder diffraction.

[0100] Example 21. Preparation of Crystal Form B of the Compound Weighed 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 0.05 mL of dichloromethane, stirred at room temperature until clarified, added 0.35 mL of ethyl acetate, stirred, slurried at room temperature for 3 days, centrifuged, and dried the solid under vacuum to obtain the product. The product was Crystal Form B detected by X-ray powder diffraction.

[0101] Example 22. Preparation of Crystal Form B of the Compound Weighed 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 0.05 mL of dichloromethane, stirred at room temperature until clarified, added 0.35 mL of acetonitrile, stirred, slurried at room temperature for 3 days, centrifuged, and dried the solid under vacuum to obtain the product. The product was Crystal Form B detected by X-ray powder diffraction.

[0102] Example 23. Preparation of Crystal Form B of the Compound Weighed 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 0.05 mL of dichloromethane, stirred at room temperature until clarified, added 0.35 mL of methyl isobutyl ketone, stirred, slurried at room temperature for 3 days, centrifuged, and dried the solid under vacuum to obtain the product. The product was Crystal Form B detected by X-ray powder diffraction.

[0103] Example 24. Preparation of Crystal Form C of the Compound Weighed 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 0.05 mL of dichloromethane, stirred at room temperature until clarified, added 0.35 mL of isopropyl acetate, stirred for crystallization, slurried at room temperature for 3 days, after centrifugation, the solid was dried in vacuo to obtain the title product. Detected by X-ray powder diffraction, the product was defined as crystalline form C, the XRD pattern was shown as in Figure 4, and its characteristic peak positions were shown as in Table 6. The DSC pattern showed that the peak values of the endothermic peaks were 86.45 °C and 255.56 °C, and the peak value of the exothermic peak was 194.44 °C. The TGA pattern showed a 5.17% weight loss from 30 °C to 140 °C.

[0104]

Table 6-1

Table 6-2

[0105] Example 25. Preparation of crystalline form D of the compound Weighed 5 mg of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, added 0.05 mL of dichloromethane, stirred at room temperature until clarified, added 0.35 mL of methyl tert-butyl ether, stirred for crystallization, slurried at room temperature for 3 days, after centrifugation, the solid was dried in vacuo to obtain the product. Detected by X-ray powder diffraction, the product was defined as crystalline form D, the XRD pattern was shown as in Figure 4, and its characteristic peak positions were shown as in Table 7. The DSC pattern showed that the peak values of the endothermic peaks were 96.46 °C and 256.21 °C, and the peak value of the exothermic peak was 160.48 °C. The TGA pattern showed a 9.49% weight loss from 30 °C to 140 °C.

[0106]

Table 7

[0107] Example 26. Preparation of Crystal Form E of the Compound 30 mg of (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was weighed, 0.15 mL of dichloromethane was added, and the mixture was stirred at room temperature until clear. Then 3.15 mL of n-heptane was added and stirred for crystallization. The mixture was slurried at room temperature for 2 days, centrifuged, and the solid was dried under vacuum to obtain the product. It was detected by X-ray powder diffraction that the product was defined as crystal form E, the XRD pattern was shown as in Figure 5, and its characteristic peak positions were shown as in Table 8. The DSC pattern showed that the peak values of the endothermic peaks were 104.97 °C, 128.14 °C, 205.60 °C, 231.49 °C, 254.86 °C, and the peak value of the exothermic peak was 184.66 °C. The TGA pattern showed a weight loss of 0.86% from 30 °C to 100 °C.

[0108]

Table 8

[0109] Experimental Example 1. Study on the Stability of Influence Factors of Crystal Form A of the Compound (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione in crystal form A was placed in an open flat, and the stability of the sample under the conditions of high temperature (40 °C, 60 °C), light irradiation, and high humidity (75% RH, 92.5% RH) was investigated. The sampling investigation period was 30 days.

[0110]

Table 9

[0111] Conclusion: From the experiments on the influencing factors, it was found that under the conditions of light irradiation, high temperatures (40°C, 60°C), and high humidities (75% RH, 92.5% RH) for 30 days, both the physical and chemical stabilities of the free crystalline form A were good.

[0112] Experimental Example 2. Long-term / Accelerated Stability Study of Crystalline Form A of the Compound The crystalline form A of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was subjected to a long-term accelerated stability study for 6 months under long-term (at 25°C and 60% RH) and accelerated (at 40°C and 75% RH) conditions.

[0113]

Table 10

[0114] Conclusion: From the long-term accelerated experiment, it was found that under the conditions of 25°C, 60% RH, and 40°C, 75% RH for 6 months, both the physicochemical stabilities of the free crystalline form A were good.

[0115] Experimental Example 3. Experiment on the Influencing Factors of Crystalline Form B of the Compound The crystalline form B of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione was placed open flat, and the stability of the sample was investigated under the conditions of high temperatures (40°C, 60°C), light irradiation, and high humidities (RH 75%, RH 90%). The sampling investigation period was 30 days.

[0116]

Table 11

[0117] Conclusion: From the experiments on the impact factor, it was found that the physicochemical stability of the free crystalline form B was good under the conditions of light irradiation, high temperature (40 °C, 60 °C), and high humidity (75% RH, 92.5% RH) for 30 days.

[0118] Experimental Example 4. Long-term / Accelerated Stability Experiment of Crystalline Form B of the Compound (S)-6-((1-(2-Fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione in crystalline form B was subjected to long-term (25 °C, 60% RH) and accelerated (40 °C, 75% RH) conditions for a 6-month long-term accelerated stability study.

[0119]

Table 12

[0120] Conclusion: From the long-term accelerated test, it was found that the physicochemical stability of the free crystalline form B was good under the conditions of 25 °C, 60% RH and 40 °C, 75% RH for 6 months.

Claims

1. Crystal form A of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ of the crystal form has characteristic peaks at 9.2, 10.9, 18.6, 20.1, 22.0 and 26.9, preferably 9. Characteristic peaks are present at 2, 10.9, 13.3, 16.9, 18.6, 19.2, 20.1, 22.0, 26.9 and 35.6, more preferably at 9.2, 10.9, 13.3, 16.9, 18.6, 19.2, 20.1, 20.6, 22.0, 26.9, 28.0 and 35.6, and most preferably, the X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 2. Crystal form A.

2. Crystal form B of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ of the crystal form has characteristic peaks at 11.1, 15.9, 18.8, 20.4, 22.2 and 26.8, preferably at 6.6 and 9. Characteristic peaks are present at 5, 11.1, 15.9, 18.8, 20.4, 21.0, 22.2, 24.6 and 26.8, more preferably at 6.6, 9.5, 11.1, 13.3, 14.0, 15.9, 18.8, 19.0, 19.8, 20.4, 21.0, 22.2, 26.8 and 28.0, and most preferably, the X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 3. Crystal form B.

3. The crystalline form C of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione has characteristic peaks at 8.3, 14.4, 16.8, 17.8, 19.3, 22.3 and 23.1, and most preferably the crystalline form C is shown in Figure 4. Crystal form C.

4. Crystal form D of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ of the crystal form has characteristic peaks at 7.8, 8.7, 9.7, 13.5, 14.0 and 22.9, preferably characteristic peaks at 7.8, 8.7, 9.7, 13.5, 14.0, 16.7, 19.6, 20.2 and 22.9, more preferably characteristic peaks at 7.8, 8.7, 9.7, 13.5, 14.0, 15.5, 16.7, 17.5, 19.6, 20.2, 22.9 and 23.9, and most preferably the X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 5. Crystal form D.

5. Crystal form E of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ of the crystal form has characteristic peaks at 6.9, 9.3, 14.2, 17.1, 18.0 and 25.0, and preferably Characteristic peaks are present at 6.9, 9.3, 13.5, 14.2, 17.1, 18.0, 22.4, 23.2 and 25.0, more preferably at 6.9, 9.3, 13.5, 14.2, 16.1, 17.1, 18.0, 21.1, 22.4, 23.2, 23.8 and 25.0, and most preferably the X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 6. Crystal form E.

6. The crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione according to any one of claims 1 to 5, wherein the error range of the 2θ angle is ±0.

2.

7. A method for preparing crystal forms A, B, C, D, or E according to any one of claims 1 to 5, comprising any one of the following methods, i.e., Method one: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with a solvent, and dissolve the compound by stirring or heating. (b) Crystallize, Alternatively, Method 2: (a) Mix the compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with a solvent, and dissolve the compound by stirring or heating. (b) Add the second solvent and crystallize, Alternatively, Method 3: (a) Mix compound (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione with a solvent, (b) Stirring to form a slurry and crystallizing, selected from the group consisting of the above, Preparation method.

8. A pharmaceutical composition comprising the following components: i) The crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione as described in any one of claims 1 to 5, and ii) Containing one or more pharmaceutically acceptable excipients, Pharmaceutical composition.

9. A method for preparing a pharmaceutical composition, comprising the step of mixing the crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione described in any one of claims 1 to 5 with a pharmaceutically acceptable excipient, method.

10. Use of the crystalline form of (S)-6-((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro-2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-dione as described in any one of claims 1 to 5 in the preparation of a drug for treating a disease or condition related to myosin regulation.

11. The aforementioned diseases or conditions include preserved ejection fraction diastolic heart failure, ischemic heart disease, angina pectoris, restrictive cardiomyopathy, diastolic dysfunction, hypertrophic cardiomyopathy (HCM), non-obstructive hypertrophic cardiomyopathy (nHCM), obstructive hypertrophic cardiomyopathy (oHCM), valvular heart disease, preserved ejection fraction heart failure (HFpEF), intermediate ejection fraction heart failure (HFmREF), aortic stenosis, inflammatory cardiomyopathy, Refrel's endocarditis, endocardial fibrosis, infiltrative cardiomyopathy, and hemochromatosis. Selected from the group consisting of Fabry disease, glycogen storage disease, congenital heart disease, Tetralogy of Fallot, left ventricular hypertrophy, refractory angina, and Chagas disease, and selectively selected from the group consisting of ischemic heart disease, restrictive cardiomyopathy, hypertrophic cardiomyopathy (HCM), non-obstructive hypertrophic cardiomyopathy (nHCM), obstructive hypertrophic cardiomyopathy (oHCM), inflammatory cardiomyopathy, infiltrative cardiomyopathy, congenital heart disease, and left ventricular hypertrophy, and selectively selected, the disease or condition is hypertrophic cardiomyopathy (HCM). Claim 10 (S)-6- ((1-(2-fluoro-5-methylphenyl)ethyl)amino)-3-(tetrahydro -2H-pyran-4-yl)-1,3,5-triazine-2,4(1H,3H)-zione The use of the crystal form.

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