Crystal of benzoxazinone compound and preparation thereof

Abstract

The invention discloses a crystal of a benzoxazinone compound and a preparation method of the crystal, and particularly discloses a compound shown as a formula (I) and a preparation method and application of the crystal of the compound. # imgabs0 #

Description

Crystallization and preparation of benzoxazinone compounds

[0001] This application claims priority to:

[0002] Application number: CN202211615935.1, application date: December 15, 2022. Technical Field

[0003] The present application relates to a crystal of a benzoxazinone compound and a preparation method thereof, and specifically discloses a preparation method and application of a compound of formula (I) and its crystal. Background Art

[0004] Aldosterone, a mineralocorticoid hormone secreted primarily by the zona glomerulosa of the adrenal cortex, plays a crucial role in regulating water and sodium balance and maintaining internal homeostasis. Recent studies have shown that excessive aldosterone can promote inflammation, myocardial remodeling, and fibrosis, and can also directly lead to renal damage and increased proteinuria. It is implicated in the development and progression of various diseases, including chronic kidney disease, hypertension, and heart failure. Mineralocorticoid receptor antagonists (MRs) block the binding of aldosterone to the mineralocorticoid receptor (MR), preventing overactivation of the aldosterone-MR complex and thereby delaying disease progression. Although early clinical trials have demonstrated that MR antagonists can improve prognosis and survival in patients with heart failure and offer renal protection beyond blood pressure, the first-generation drug, spironolactone, has poor nuclear receptor selectivity, and long-term use can lead to adverse effects such as gynecomastia, impotence, and menstrual irregularities in women. The second-generation drug, eplerenone, while offering improved nuclear receptor selectivity, is weak and difficult to synthesize. Both spironolactone and eplerenone carry the risk of hyperkalemia in clinical use, limiting the application of MR antagonists.

[0005] Summary of the Invention

[0006] In one aspect, the present application provides a crystal of a compound of formula (I),

[0007] In some embodiments of the present application, the crystal of the compound of formula (I) of the present application is crystal A, and its X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 13.88±0.20° and 15.59±0.20°.

[0008] In some embodiments of the present application, the above-mentioned A crystal has an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.52±0.20° and 25.92±0.20°.

[0009] In some embodiments of the present application, the above-mentioned A crystal has an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20° and 25.92±0.20°.

[0010] In some embodiments of the present application, the above-mentioned A crystal has an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 15.99±0.20°, 16.59±0.20°, 18.35±0.20°, 18.60±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20°, 25.92±0.20° and 27.10±0.20°.

[0011] In some embodiments of the present application, the X-ray powder diffraction pattern of the above-mentioned crystal A, expressed as 2θ values, comprises 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15 or 16 diffraction peaks selected from the following: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0. 20°, 15.59±0.20°, 15.99±0.20°, 16.59±0.20°, 18.35±0.20°, 18.60±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20°, 25.92±0.20° and 27.10±0.20°.

[0012] In some embodiments of the present application, the above-mentioned A crystal has an X-ray powder diffraction pattern, represented by 2θ values, comprising 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 diffraction peaks selected from the following: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20° and 25.92±0.20°.

[0013] In some embodiments of the present application, the above-mentioned A crystal, in its X-ray powder diffraction pattern, expressed as 2θ values, contains 3, 4, 5, 6, 7 or 8 diffraction peaks selected from the following: 10.52±0.20°, 11.45±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.52±0.20° and 25.92±0.20°.

[0014] In some embodiments of the present application, the above-mentioned crystal A has an X-ray powder diffraction pattern with diffraction peaks at the following 2θ angles: 6.20±0.20°, 9.04±0.20°, 9.60±0.20°, 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 12.82±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 15.99 ±0.20°, 16.59±0.20°, 17.18±0.20°, 17.54±0.20°, 18.35±0.20°, 18.60±0.20°, 19.22±0.20°, 19.52±0.20°, 20.03±0.20°, 20.29±0.20°, 20.96±0.20°, 21.11±0.20°, 21.57±0.20°, 22.23±0.20° , 23.08±0.20°, 23.33±0.20°, 23.82±0.20°, 24.22±0.20°, 24.86±0.20°, 25.05±0.20°, 25.26±0.20°, 25.52±0.20°, 25.92±0.20°, 27.10±0.20°, 27.33±0.20°, 27.64±0.20°, 28.82±0.20°, 29.20 ±0.20°, 29.68±0.20°, 30.21±0.20°, 30.59±0.20°, 31.19±0.20°, 31.75±0.20°, 32.52±0.20°, 33.71±0.20°, 34.51±0.20°, 35.88±0.20°, 36.54±0.20°, 37.45±0.20°, 38.23±0.20° and 38.60±0.20°.

[0015] In some embodiments of the present application, the above-mentioned crystal A has an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 6.20°, 9.04°, 9.60°, 10.52°, 11.45°, 12.38°, 12.82°, 13.21°, 13.88°, 15.59°, 15.99°, 16.59°, 17.18°, 17.54°, 18.35°, 18.60°, 19.22°, 19.52°, 20.03°, 20.29°, 20.96°, 21.11°, 21.57° , 22.23°, 23.08°, 23.33°, 23.82°, 24.22°, 24.86°, 25.05°, 25.26°, 25.52°, 25.92°, 27.10°, 27.33°, 27.64°, 28.82°, 29.20°, 29.68°, 30.21°, 30.59°, 31.19°, 31.75°, 32.52°, 33.71°, 34.51°, 35.88°, 36.54°, 37.45°, 38.23° and 38.60°.

[0016] In some embodiments of the present application, the above-mentioned A crystal has an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 13.88±0.20°, and / or 15.59±0.20°, and / or 6.20±0.20°, and / or 9.04±0.20°, and / or 9.60±0.20°, and / or 12.38±0.20°, and / or 12.82±0.20°, and / or 13.21±0.20°, and / or 15.99±0.20°, and and / or 16.59±0.20°, and / or 17.18±0.20°, and / or 17.54±0.20°, and / or 18.35±0.20°, and / or 18.60±0.20°, and / or 19.22±0.20°, and / or 19.52±0.20°, and / or 20.03±0.20°, and / or 20.29±0.20°, and / or 20.96±0.20°, and / or 21.11±0.20°, and / or 21.57±0.20°, and / or 22.23±0.20°, and / or 23.08±0.20°, and / or 23.33±0.20°, and / or 23.82±0.20°, and / or 24.22±0.20°, and / or 24.86±0.20°, and / or 25.05±0.20°, or 25.26±0.20°, and / or 25.52±0.20°, and / or 25.92±0.20°, and / or 27.10±0.20°, and / or 27.33±0.20°, and / or 27.64±0.20°, and / or 28.82±0.20°, and / or 29.20 ±0.20°, and / or 29.68±0.20°, and / or 30.21±0.20°, and / or 30.59±0.20°, and / or 31.19±0.20°, and / or 31.75±0.20°, and / or 32.52±0.20°, and / or 33.71±0.20°, and / or 34.51±0.20°, and / or 35.88±0.20°, and / or 36.54±0.20°, and / or 37.45±0.20°, and / or 38.23±0.20°, and / or 38.60±0.20°.

[0017] In some embodiments of the present application, the peak position, intensity, interplanar spacing and relative intensity of the diffraction peaks in the X-ray powder diffraction pattern of the above-mentioned crystal A are shown in Table 1 below:

[0018] Table 1 XRPD pattern analysis data of compound A of formula (I)

[0019] In some embodiments of the present application, the XRPD pattern of the above-mentioned crystal A is basically as shown in Figure 1.

[0020] In some embodiments of the present application, the above-mentioned A crystal has a differential scanning calorimetry curve (DSC graph) having an endothermic peak starting point at 215.9°C±5°C.

[0021] In some embodiments of the present application, the above-mentioned crystal A has a DSC spectrum substantially as shown in FIG2 .

[0022] In some embodiments of the present application, the thermogravimetric analysis (TGA) curve of the above-mentioned crystal A shows a weight loss of 0.90% at 200°C±3°C.

[0023] In some embodiments of the present application, the TGA spectrum of the above-mentioned crystal A is basically as shown in Figure 3.

[0024] In some embodiments of the present application, the above-mentioned A crystal has a moisture absorption weight increase of greater than 0.2% and less than 2% at 25° C. and 80% relative humidity.

[0025] In some embodiments of the present application, the water adsorption isotherm (DVS) spectrum of the above-mentioned A crystal is basically as shown in Figure 4.

[0026] On the other hand, the present application also provides a method for preparing the above-mentioned A crystal, which comprises the following steps:

[0027] (1) dissolving the compound of formula (I) in a crystallization solvent;

[0028] (2) Crystallization, and then separation of the solid to obtain crystal A of the compound of formula (I).

[0029] In some embodiments of the present application, the crystallization solvent in step (1) is selected from dichloromethane, tetrahydrofuran, dioxane, acetone, acetonitrile, ethanol, methanol, water or a mixed solvent of any two thereof.

[0030] In some embodiments of the present application, the crystallization solvent in step (1) is selected from tetrahydrofuran, acetone, acetonitrile, water or a mixed solvent of any two of these solvents.

[0031] In some embodiments of the present application, the crystallization solvent in step (1) is selected from tetrahydrofuran, acetone, a mixed solvent of acetonitrile and water, and a mixed solvent of acetone and water.

[0032] On the other hand, the present application also provides a single crystal of the compound of formula (I).

[0033] In some embodiments of the present application, the compound of formula (I) is a single crystal, and the single crystal has the following characteristics: it belongs to the orthorhombic system, and the unit cell parameters are α=90°, β=90°, γ=90°, Z=8, crystal density is 1.439 mg / m 3 .

[0034] In some embodiments of the present application, the compound of formula (I) is a single crystal, and the absolute configuration of the single crystal of the compound of formula (I) is S configuration.

[0035] In some embodiments of the present application, the compound of formula (I) is a single crystal, and the ellipsoidal diagram of the single crystal molecular structure is shown in FIG5 .

[0036] In another aspect, the present application also provides a crystalline composition, wherein the crystal A of the compound of formula (I) accounts for at least 50% by weight of the crystalline composition, preferably at least 75%, more preferably at least 90%, and most preferably at least 95%. The crystalline composition may also contain a small amount of other crystalline or amorphous forms of the compound of formula (I). The crystalline composition may also contain a small amount of other crystalline or amorphous forms of the compound of formula (I), or impurities other than these substances.

[0037] On the other hand, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of crystal A of the compound of formula (I) above, or a crystalline composition of the compound of formula (I) above; the pharmaceutical composition may comprise at least one pharmaceutically acceptable carrier or other excipient.

[0038] In some embodiments of the present application, the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is selected from pharmaceutical compositions for oral administration, subcutaneous administration, intramuscular administration, or intravenous administration.

[0039] In some embodiments of the present application, the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is selected from pharmaceutical compositions for oral administration, intramuscular administration, or intravenous administration.

[0040] In some embodiments of the present application, the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is administered via a solid pharmaceutical composition.

[0041] In some embodiments of the present application, the solid pharmaceutical composition is selected from tablets or capsules.

[0042] In some embodiments of the present application, the dosage of the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is 0.0001 mg / kg to 100 mg / kg.

[0043] In some embodiments of the present application, the dosage of the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is 0.001 mg / kg to 80 mg / kg.

[0044] In some embodiments of the present application, the dosage of the crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I), or the pharmaceutical composition is 0.01 mg / kg to 60 mg / kg.

[0045] On the other hand, the present application also provides the use of crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I) or the pharmaceutical composition in the preparation of drugs for treating diseases related to mineralocorticoid receptor antagonists.

[0046] On the other hand, the present application also provides the use of crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I) or the pharmaceutical composition in the preparation of a drug for treating diabetic nephropathy.

[0047] On the other hand, the present application also provides a method for treating a disease associated with a mineralocorticoid receptor antagonist in a subject in need thereof, comprising providing the subject with an effective dose of crystal A of the compound of formula (I), a crystalline composition of the compound of formula (I), or the pharmaceutical composition.

[0048] On the other hand, the present application also provides the use of crystal A of the compound of formula (I), the crystalline composition of the compound of formula (I) or the pharmaceutical composition in the treatment of diseases associated with mineralocorticoid receptor antagonists.

[0049] On the other hand, the present application also provides crystal A of the compound of formula (I), a crystalline composition of the compound of formula (I), or the pharmaceutical composition for treating diseases associated with mineralocorticoid receptor antagonists.

[0050] In some embodiments of the present application, the above-mentioned mineralocorticoid receptor antagonist-related diseases are selected from diabetic nephropathy.

[0051] Technical Effects

[0052] The compounds of this application have excellent antagonistic activity against mineralocorticoid receptors. Furthermore, the compounds exhibit excellent pharmacokinetic and pharmacodynamic properties, as well as good membrane permeability and solubility. The crystals of the compounds of this application are stable, hygroscopic, and minimally affected by light and heat, suggesting promising drug development prospects.

[0053] Definition and Description

[0054] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be construed as ambiguous or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding commercial product or its active ingredient.

[0055] Unless otherwise stated, X-ray powder diffraction (XRPD) can detect information such as changes in crystallinity, crystallinity, and crystalline state, and is a common means of identifying crystals. The peak position of the XRPD pattern depends primarily on the structure of the crystal and is relatively insensitive to experimental details, while its relative peak height depends on many factors related to sample preparation and instrument geometry. Therefore, for any given crystalline form, the relative intensity of the diffraction peak can change due to preferred orientation caused by factors such as crystal morphology, which is well known in the art of crystallography. Where there is a preferred orientation effect, the peak intensity changes, but the diffraction peak position of the crystal cannot be changed. In addition, for any given crystal, there may be slight errors in the position of the peak, which is also well known in the art of crystallography. For example, due to changes in temperature during sample analysis, movement of the sample, or calibration of the instrument, the position of the peak can move, and the measurement error of the 2θ value is sometimes about ±0.2 degrees. The measurement of 2θ of the XRPD pattern may vary slightly between different instruments and different samples, so the numerical value of the 2θ cannot be considered absolute. Therefore, it is well known to those skilled in the art that this error should be taken into account when determining each crystalline structure. Therefore, in some embodiments, the crystals of the present application are characterized by an XRPD pattern with certain peak positions, which is substantially as shown in the XRPD pattern provided in the accompanying drawings of the present application.

[0056] DSC measures the transition temperatures of crystals when they absorb or release heat due to changes in their crystalline structure or melting. For the same crystals of the same compound, the error in thermal transition temperatures and melting points in consecutive analyses is typically within about 5°C. When we say a compound has a given DSC peak or melting point, this refers to the DSC peak or melting point ±5°C. DSC provides an auxiliary method for distinguishing different crystals. Different crystalline forms can be identified by their different transition temperature characteristics. It should be noted that for mixtures, their DSC peaks or melting points may vary over a wider range. In addition, because decomposition is associated with the melting process of a substance, the melting temperature is related to the heating rate.

[0057] The term "treatment" means administering the compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0058] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0059] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0060] The term "prevention" means administering a compound or formulation described herein to prevent one or more symptoms associated with the disease, and includes preventing the occurrence of a disease or disease state in a mammal, particularly when such mammal is susceptible to the disease state but has not yet been diagnosed as having the disease state.

[0061] Reference throughout this specification to "one embodiment" or "an embodiment" or "in another embodiment" or "in certain embodiments" means that the specific referenced elements, structures, or features described in connection with that embodiment are included in at least one embodiment. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" or "in another embodiment" or "in certain embodiments" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the specific elements, structures, or features may be combined in any suitable manner in one or more embodiments.

[0062] It should be understood that the singular article "a," "an," and "the" as used in this specification and the appended claims includes plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to a reaction including "a catalyst" includes one catalyst, or two or more catalysts. It should also be understood that the term "or" is generally used in its sense including "and / or" unless the context clearly dictates otherwise.

[0063] The intermediate compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0064] The chemical reactions described in the specific embodiments of the present application are carried out in a suitable solvent that is compatible with the chemical transformations described herein and the reagents and materials required. To obtain the compounds described herein, it may sometimes be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0065] The present application will be described in detail below through examples, which are not intended to limit the present application in any way.

[0066] All solvents used in this application were commercially available and used without further purification.

[0067] The solvents used in this application are commercially available.

[0068] This application uses the following abbreviations:

[0069] N2: nitrogen; RH: relative humidity; mL: milliliter; L: liter; min: minute; ℃: degree Celsius; μm: micrometer; mm: millimeter; μL: microliter; moL / L: mole per liter; mg: milligram; s: second; nm: nanometer; MPa: megapascal; lux: lux; μw / cm2: microwatt per square centimeter; h: hour; Kg: kilogram; nM: nanomole; RT: retention time; RRT: relative retention time; rpm: rotational speed; Kv: kilovolt; mA: milliampere; DMF stands for N,N-dimethylformamide.

[0070] Instruments and analytical methods

[0071] 1. X-ray powder diffractometer (XRPD) method of this application. Test parameters are shown in Table 2.

[0072] Table 2 XRPD test parameters

[0073] 2. The differential scanning calorimeter (DSC) method of this application, the test parameters are shown in Table 3.

[0074] Table 3 DSC test parameters

[0075] 3. Thermogravimetric analysis (TGA) method of this application. Test parameters are shown in Table 4.

[0076] Table 4 TGA test parameters

[0077] 4. The dynamic vapor sorption analysis (DVS) method of this application, the test parameters are shown in Table 5.

[0078] Table 5 DVS test parameters BRIEF DESCRIPTION OF THE DRAWINGS

[0079] FIG1 is a Cu-Kα radiation XRPD spectrum of a crystal of compound A of formula (I);

[0080] FIG2 is a DSC spectrum of a crystal of compound A of formula (I);

[0081] FIG3 is a TGA spectrum of the crystals of compound A of formula (I);

[0082] FIG4 is a DVS spectrum of a crystal of compound A of formula (I);

[0083] FIG5 is an ellipsoid diagram of the three-dimensional structure of a single crystal of the compound of formula (I). DETAILED DESCRIPTION

[0084] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any adverse way. The compound of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining the specific embodiments with other chemical synthesis methods, and equivalent replacement modes well known to those skilled in the art, and preferred embodiments include but are not limited to the examples of the present invention. It will be apparent to those skilled in the art that various changes and improvements will be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0085] Example 1: Preparation of Intermediate 1

[0086] Synthesis line:

[0087] first step

[0088] Compound 1-1 (25 g, 203 mmol) and potassium carbonate (84.2 g, 609 mmol) were dissolved in N,N-dimethylformamide (200 mL). Chloroacetyl chloride (17.0 mL, 213 mmol) was added at 0°C and the mixture was reacted at 20°C for 2 hours. Water (1.5 L) was added to the reaction solution, stirred for 20 minutes, and then filtered. The filter cake was dried to obtain compound 1-2. MS-ESI calculated value [M+H] + 164, measured value 164.

[0089] Step 2

[0090] Compound 1-2 (25.6 g, 157 mmol) was dissolved in N,N-dimethylformamide (250 mL), and N-bromosuccinimide (30.7 g, 173 mmol) was added. The mixture was stirred at 20°C for 0.5 hours. Water (1500 mL) was added to the reaction mixture and stirred for 30 minutes. The mixture was filtered and the filter cake was dried under reduced pressure to obtain compound 1-3. MS-ESI calculated value [M+H] + 242, 244, measured values ​​242, 244.

[0091] Step 3

[0092] Compound 1-3 (170 g, 702.28 mmol), compound 1-(tributyltin)-methanol (315.69 g, 983.19 mmol), chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (16.58 g, 21.07 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (20.09 g, 42.14 mmol) were dissolved in dioxane (1360 mL), and the reaction solution was replaced with nitrogen three times. The reaction solution was stirred at 80 ° C. under a nitrogen atmosphere for 16 hours. The reaction solution was cooled to 25 ° C. 6.2 L of n-heptane was added to the reaction solution, filtered, and the filter cake was slurried with 1 L of n-heptane and filtered to obtain intermediate 1. MS-ESI calculated value [M+H] + 194 Actual measured value 194. 1 H NMR(400MHz, DMSO-d6)δ =10.17 (s, 1H), 6.93 (d, J = 8.0Hz, 1H), 6.77 (d, J = 8.4Hz, 1H), 4.97 (t, J = 5.4Hz, 1H), 4.46 (s, 2H), 4.41 (d, J = 5.6Hz, 2H), 2.16 (s, 3H).

[0093] Example 2: Preparation of Intermediate 2

[0094] Synthesis line:

[0095] first step

[0096] Compound 2-1 (1.00 g, 5.71 mmol) was dissolved in ethanol (5 mL), and palladium / carbon (100 mg, 10% purity) was added. The reaction mixture was stirred at 20°C under a hydrogen atmosphere (15 psi) for 4 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-2. MS-ESI calculated value [M+H] + 146, measured value 146.

[0097] Step 2

[0098] Compound 2-2 (820 mg, 5.65 mmol) and ethyl 4-chloroacetoacetate (1.12 g, 6.78 mmol) were dissolved in tetrahydrofuran (10 mL), heated to 50 ° C, and triethylamine (787 μL, 5.65 mmol) was added to the reaction solution and stirred at 50 ° C for 4 hours. After the reaction was completed, ethyl acetate (50 mL) was added to the reaction solution for extraction. The organic phase was washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by column chromatography (eluent: petroleum ether / ethyl acetate, 100 / 1 to 10 / 1, V / V) to obtain compound 2-3. MS-ESI calculated value [M+H] + 256, measured value 256.

[0099] Step 3

[0100] Compound 2-3 (500 mg, 1.96 mmol) was dissolved in ethanol (5 mL), and palladium / carbon (100 mg, 10% purity) was added to the reaction solution. The reaction mixture was stirred under a hydrogen atmosphere (15 psi) at 20°C for 14 hours. The reaction solution was filtered and concentrated under reduced pressure. The resulting crude product was redissolved in ethanol (10 mL), and palladium / carbon (200 mg, 10% purity) was added. The reaction mixture was further stirred under a hydrogen atmosphere (15 psi) at 20°C for 14 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2-4. MS-ESI calculated value [M+H] + 258, measured value 258.

[0101] Step 4

[0102] Compound 2-4 (5.5 g, 21.38 mmol) was dissolved in methanol (60 mL), and a solution of lithium hydroxide monohydrate (1.35 g, 32.07 mmol) dissolved in water (15 mL) was added to the reaction solution. The reaction mixture was stirred at 20°C for 12 hours. After the reaction was completed, the methanol was removed by concentration under reduced pressure, and water (50 mL) was added to dilute the mixture. The pH was adjusted to 4 with aqueous hydrochloric acid (2 M), and the mixture was extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 2-5. MS-ESI calculated value [M+H] + 230, measured value 230.

[0103] Step 5

[0104] Compound 2-5 (64 g, 279.26 mmol) was dissolved in acetonitrile (960 mL), and compound 2-6 (59.01 g, 279.26 mmol) was added. The reaction mixture was stirred at 25 ° C for 0.5 hours. The reaction mixture was heated to 70 ° C and stirred for 1 hour. The temperature was slowly lowered to 25 ° C and stirred at 25 ° C for 12 hours. The reaction solution was filtered, the filter cake was washed with acetonitrile (32 mL * 2), and the filter cake was collected. The filter cake was added to the reaction flask, 320 mL of water was added, the pH was adjusted to 10-11 with 1M sodium hydroxide aqueous solution, and extracted 3 times with ethyl acetate (320 mL * 3). The aqueous phase was adjusted to pH 2-3 with 12M hydrochloric acid solution and extracted twice with ethyl acetate (320 mL * 2). The combined organic phase was washed with saturated brine (640 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 2-7. MS-ESI calculated value [M+H] + 230, measured value 230.

[0105] Step 6

[0106] Compound 2-7 (48 g, 209.44 mmol) was dissolved in N,N-dimethylformamide (240 mL). 1,1'-Carbonyldiimidazole (44.15 g, 272.28 mmol) was added to the reaction solution, and the reaction mixture was stirred at 25°C for 2 hours. Ammonium chloride (33.61 g, 628.33 mmol) and N,N-diisopropylethylamine (40.60 g, 314.16 mmol, 54.72 mL) were then added to the reaction solution, and the reaction mixture was stirred at 25°C for 2 hours. The reaction solution was diluted with water (480 mL) and extracted with ethyl acetate (480 mL x 4). The combined organic phases were washed once with 1M hydrochloric acid (0.96 L) and then with saturated brine (0.96 L x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was dissolved in ethyl acetate (90 mL), and n-heptane (450 mL) was added and stirred at 25°C for 0.5 hours. The mixture was filtered, and the filter cake was washed with n-heptane (90 mL*3) and dried under reduced pressure to obtain intermediate 2. MS-ESI calculated value [M+H] + 229, measured value 229.

[0107] Example 3: Preparation of compound of formula (I)

[0108] Synthesis route:

[0109] first step

[0110] Intermediate 1 (52 g, 269.15 mol) and trimethylsilyl chloride (58.48 g, 538.31 mol) were dissolved in dichloromethane (234 mL) and DMSO (5.26 g, 67.29 mmol) was slowly added dropwise. The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was filtered, and the filter cake was rinsed with dichloromethane (52 mL) and dried to obtain crude compound 3. 1 H NMR (400MHz, DMSO-d6) δ = 10.28 (s, 1H), 7.02 (d, J = 8.2Hz, 1H), 6.81 (d, J = 8.2Hz, 1H), 4.76 (s, 2H), 4.51 (s, 2H), 2.26 (s, 3H).

[0111] Step 2

[0112] Compound 3 (37 g, 174.82 mmol) and intermediate 2 (38.70 g, 169.58 mmol) were dissolved in acetone (222 mL). Sodium iodide (5.24 g, 34.96 mmol) and sodium bicarbonate (15.42 g, 183.56 mmol) were added to the reaction mixture, and the reaction mixture was stirred at 50°C for 16 hours. Water (666 mL) was added to the reaction mixture in three batches, and the mixture was stirred at 20-30°C for 0.5-1 hour. The reaction mixture was filtered, and the filter cake was washed with water (74 mL*3) until the pH of the aqueous phase reached 7. The crude product was added to water (370 mL) at 20-30°C and stirred for 0.5-1 hour. The mixture was filtered, and the filter cake was washed with water (74 mL*3) and dried under vacuum. The filter cake was dried under reduced pressure to obtain the crude product. The crude product (62.5 g, 154.94 mmol) was dissolved in tetrahydrofuran (437.5 mL), heated to 65°C, and stirred for 2 hours. Filtered while hot, the filtrate was cooled to 50-60°C, and methyl tert-butyl ether (647.85 g, 7.35 mol, 875.00 mL) was slowly added at 50-60°C. Stirred at 50-60°C for 1 hour. The temperature was slowly lowered to 20°C and stirred at 20°C for 12 hours. Filtered, the filter cake was washed with methyl tert-butyl ether (125 mL x 3), and dried under reduced pressure to yield the compound of formula (I). The ee value of the compound was determined by SFC (Chromatographic column: Chiralpak IG-3 50×4.6 mm ID, 3 μm; mobile phase: supercritical CO2-0.05% ethylene glycolamine in ethanol; gradient: 0.05% ethylene glycolamine in ethanol: 5%-40%). MS-ESI calculated value [M+H] + 404, found 404. ee value = 100%, RT = 1.196 min. 1H NMR (400MHz, DMSO-d6) δ = 10.19 (s, 1H), 7.23 (s, 1H), 7.06 (d, J = 8.4Hz, 1H), 6.90-6.75 (m, 3H), 6.67 (m, 1H), 4.50 (s, 2 H), 4.19 (d, J=14.8Hz, 1H), 4.13-3.89 (m, 3H), 3.34-3.29 (m, 1H), 2.24 (s, 3H), 2.17-2.05 (m, 1H), 2.04-1.93 (m, 1H).

[0113] Example 4: Preparation of crystals of compound A of formula (I)

[0114] 50 mg of the compound of formula (I) was weighed and added to a glass vial, followed by an appropriate amount of tetrahydrofuran. After adding a magnetic stirrer, the sample was placed at room temperature (25°C) and magnetically stirred overnight. The sample was filtered, and the solid sample on the filter cake was dried in a vacuum drying oven (45°C) overnight to obtain crystals of compound (I) A. The XRPD, DSC, and TGA results of crystals of compound (I) A are shown in Figures 1, 2, and 3, respectively.

[0115] The above tetrahydrofuran was replaced with the solvents listed in Table 6, and the same operation was performed as above. The results are shown in Table 6.

[0116] Table 6 Room temperature stirring test

[0117] Experimental Example 1: Hygroscopicity Study of Crystal A of Compound of Formula (I)

[0118] Experimental Materials:

[0119] SMS DVS intrinsic dynamic water vapor sorption instrument

[0120] Experimental methods:

[0121] 10-30 mg of crystals of compound A of formula (I) were placed in a DVS sample tray for testing.

[0122] The classification of moisture absorption evaluation is shown in Table 7.

[0123] Table 7 Moisture absorption evaluation classification table

[0124] Note: ΔW% indicates the weight gain of the test sample at 25±1℃ and 80±2%RH.

[0125] Experimental results:

[0126] The DVS spectrum of crystal A of compound (I) is shown in Figure 4. The sample was tested using a 0-95-0% RH / 25°C cycle, with the sample pre-equilibrated at 0% RH. Compared to the initial humidity of 0%, the sample gained 0.2976% (cyclic adsorption) when the humidity rose to 80%, indicating that the sample is slightly hygroscopic. XRPD analysis revealed no change in the crystal structure before and after DVS testing.

[0127] Experimental Example 2: Solid Stability Test of Compound A of Formula (I)

[0128] According to the Guiding Principles for Stability Testing of APIs and Pharmaceutical Preparations (Chinese Pharmacopoeia 2015 Edition, Part IV, General Rules 9001), the stability of the crystals of compound A of formula (I) was investigated under high temperature (60°C, open), high humidity (room temperature / relative humidity 92.5%, open) and light (total illumination = 1.2×10 6 Lux·hr / Near UV=200w·hr / m 2 , stability under open) conditions.

[0129] Weigh 10 mg of crystals of compound A of formula (I) and place them at the bottom of a glass sample bottle, spreading them into a thin layer. For samples stored under high temperature (60°C) and high humidity (92.5% RH), seal the bottle with aluminum foil and poke small holes in the foil to ensure sufficient contact with ambient air. Place the sample in a corresponding constant temperature and humidity chamber. The illuminated sample (open, not covered with aluminum foil) and the illuminated control (the entire sample bottle was covered with aluminum foil) were placed in the illumination chamber. Weigh two portions at each time point to serve as the official test samples. Separately, weigh approximately 50 mg of crystals of compound A of formula (I) for XRPD analysis. The sample bottle was wrapped in aluminum foil and poke small holes in the same chamber. Samples were taken for XRPD analysis on days 5 and 10, and the results were compared with the initial test results on day 0. The results of the solid-state pre-stability test of crystals of compound A of formula (I) are shown in Table 8.

[0130] Table 8 Results of solid pre-stability test of crystal A of compound of formula (I)

[0131] Experimental conclusion: Crystal A of the compound of formula (I) has good stability under high temperature, high humidity and strong light conditions.

[0132] Experimental Example 3: Single crystal X-ray diffraction analysis of the compound of formula (I)

[0133] Instrument parameters

[0134] Instrument: Bruker D8 VENTURE single crystal X-ray diffractometer; cryogenic system: Oxford Cryostream 800; light source: Cu rotating target, 2.5kW; sample to detector distance: d = 45mm; tube voltage: 50kV; tube current: 45mA.

[0135] Crystal cultivation

[0136] 10 mg of the compound of formula (I) was added to 4 mL of methanol and stirred until the sample partially dissolved. 1 mL of DMF was then added and stirred until the sample was completely dissolved. The clear sample solution was placed in a 4 mL semi-sealed sample vial and slowly evaporated at room temperature. Colorless blocky crystals were obtained after two weeks.

[0137] Data collection and tabulation

[0138] The single crystal is the compound of formula (I). The crystal was collected and diffraction intensity data was collected using a Bruker D8 VENTURE single crystal X-ray diffractometer. The crystal structure data of the compound of formula (I) are shown in Tables 9 to 14.

[0139] Table 9 Single crystal structure data of compound of formula (I)

[0140] Table 10 Atomic coordinates of single crystal of compound of formula (I) (×10 4 ) and the equivalent isotropic shift parameter

[0141] Table 11 Bond lengths of single crystals of compound of formula (I)

[0142] Table 12 Bond angles of single crystals of compound of formula (I)

[0143] Table 13 Twist angle of single crystal of compound of formula (I)

[0144] Table 14 Hydrogen atom coordinates of single crystal of compound of formula (I) (×10 4 ) and isotropic displacement parameters

[0145] Experimental Conclusion

[0146] The single crystal data showed that the single crystal was the compound of formula (I), and the absolute configuration of the compound of formula (I) was confirmed to be S. The three-dimensional structure ellipsoid diagram of the single crystal of the compound of formula (I) is shown in Figure 5.

[0147] Experimental Example 4: In vitro evaluation of MR antagonist activity

[0148] Experimental purpose: to test the MR antagonist activity of the compound

[0149] Experimental Materials:

[0150] DMEM medium was purchased from BI; fetal bovine serum was purchased from Biosera; HEK293 / Gal4 / MR cell line was provided by Wuhan Heyan Biopharmaceutical Technology Co., Ltd.; Bright Glo was purchased from Promega; EnVision multi-label analyzer was purchased from PerkinElmer.

[0151] Experimental methods:

[0152] MR cells were seeded in a white 96-well plate at a density of 40,000 cells per well in 80 ml of cell suspension. The plate was incubated in a CO2 incubator overnight.

[0153] The compound to be tested was diluted with a dispenser to 8 concentration points, with a 5-fold gradient dilution, and the compound concentration was 2 millimolar to 0.026 micromolar. 38 microliters of culture medium was added to the middle plate, and then 2 microliters of compound per well were added to the middle plate according to the corresponding position. After mixing, 10 microliters of compound solution per well were transferred to the cell plate, and the cell plate was placed in a carbon dioxide incubator and incubated for 1 hour. Aldosterone was diluted to 10 nanomolar using culture medium, that is, 20 microliters of aldosterone with a concentration of 1 micromolar was added to 1980 microliters of culture medium, and after mixing, 10 microliters of aldosterone solution was added to each well of the cell plate except the positive control well. The cell plate was placed in a carbon dioxide incubator and incubated for 24 hours. The final concentration of the compound was 10 micromolar to 0.128 nanomolar, and the final concentration of aldosterone was 1 nanomolar.

[0154] After incubation, discard the cell supernatant, add 100 μl of BrightGlo reagent per well to the cell plate, and read immediately on the EnVision Multilabel Analyzer.

[0155] Data Analysis:

[0156] Using the equation %Inhibition=((RFU Cmpd -AVER(RFU Neg.Ctrl )) / ((AVER(RFU Pos.Ctrl )-AVER(RFU Neg.CtrlThe raw data were converted into inhibition rate by multiplying by 100%, and the compound curve was fitted by log(inhibitor) vs. response-Variable slope in Graphpad Prism 5. IC 50 The value is calculated by the software using the formula Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)).

[0157] Experimental results: See Table 15.

[0158] Table 15 MR antagonistic activity test results

[0159] Experimental conclusion: The compound of the present application has good antagonistic activity against mineralocorticoid receptors.

[0160] Experimental Example 5: Pharmacokinetic Evaluation

[0161] Experimental Materials:

[0162] SD rats (male, 7-9 weeks old, Beijing Weitonglihua)

[0163] Experimental operation:

[0164] The pharmacokinetic profile of the compound in rodents was tested following intravenous (IV) and oral (PO) administration using standard protocols. Rats were given single IV and PO doses. The IV vehicle was water. The IV vehicle consisted of a clear solution of 80% polyethylene glycol 400 / 20% water, while the oral vehicle was a homogenous suspension of 99.9% (0.5% hydroxypropyl methylcellulose: polyethylene glycol stearate 15 = 95:5) / 0.1% Tween 80. Four male Sprague-Dawley rats were used in this study. Two rats were dosed intravenously, and plasma samples were collected at 0 h (pre-dose) and 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 h post-dose. Two rats were dosed orally by gavage, and plasma samples were collected at 0 h (pre-dose) and 0.25, 0.5, 1, 2, 4, 8, and 24 h post-dose. Whole blood samples were also collected over a 24-hour period. All blood samples were immediately transferred to labeled commercial centrifuge tubes containing K2-EDTA. After blood sample collection, the supernatant plasma was aspirated at 3200 g for 10 minutes at 4°C, immediately placed on dry ice, and then stored at -60°C or lower for LC-MS / MS analysis. The non-compartmental model was used to analyze the plasma concentration-time data using the WinNonlin software package (Version 6.3 and above) and to calculate pharmacokinetic parameters such as peak concentration (C max ), clearance (CL), tissue distribution (Vdss), area under the drug-time curve (AUC 0-last), bioavailability (F), etc.

[0165] The experimental results are shown in Table 16.

[0166] Table 16 Pharmacokinetic evaluation results of the compounds of the present application in rats

[0167] Experimental conclusion: The compound of the present application has good pharmacokinetic properties.

[0168] Experimental Example 6: In vivo efficacy test in unilateral nephrectomized rats

[0169] Experimental Materials:

[0170] SD rats (male, Beijing Weitonglihua Laboratory Animal Technology Co., Ltd.)

[0171] Experimental operation:

[0172] After 2-3 days of adaptive feeding, male SD rats underwent right nephrectomy and were fed a standard diet and drinking water. After one week of recovery, rats were divided into groups according to body weight and subcutaneously implanted with an osmotic pump. Aldosterone was injected at a concentration of 3 mg / mL (dissolved in 0.15% DMSO / prepared with sterile water) (Source Leaf Biotechnology S30644-5 mg) at a flow rate of 0.75 μg / hr (Alzet Model 2004). The rats were also fed a 6% NaCl high-salt diet (customized by Beijing Keao Xieli Feed Co., Ltd.) and 0.3% KCl in drinking water (Source Leaf Biotechnology S24120). Oral administration was performed simultaneously with the subcutaneous osmotic pump implantation. The dosage was detailed in the grouping. The oral administration vehicle (0.5% hydroxypropyl methylcellulose, polyethylene glycol stearate 15 = 95:5 (v / v, containing 0.1% Tween 80)) was administered once daily for 4 consecutive weeks. Body weight was measured and recorded daily. After 4 weeks of administration, 24-hour urine was collected to detect urine albumin and urine creatinine, and the urine albumin to urine creatinine ratio (UACR) was calculated.

[0173] Experimental conclusion: The compound of the present application has a significant protective effect on the kidneys and can effectively reduce the ratio of albumin to creatinine in rat urine.

Claims

1. A crystal of the compound of formula (I), having an X-ray powder diffraction pattern having diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 13.88±0.20°, and 15.59±0.20°; 2. The crystal according to claim 1, wherein the X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.52±0.20° and 25.92±0.20°.

3. The crystal according to claim 2, wherein the X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 16.59±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20° and 25.92±0.20°.

4. The crystal according to claim 3, wherein the X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 15.99±0.20°, 16.59±0.20°, 18.35±0.20°, 18.60±0.20°, 19.22±0.20°, 19.52±0.20°, 23.82±0.20°, 25.05±0.20°, 25.92±0.20° and 27.10±0.20°.

5. The crystal according to claim 4, wherein the X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 6.20±0.20°, 9.04±0.20°, 9.60±0.20°, 10.52±0.20°, 11.45±0.20°, 12.38±0.20°, 12.82±0.20°, 13.21±0.20°, 13.88±0.20°, 15.59±0.20°, 15.99±0.20° .20°, 16.59±0.20°, 17.18±0.20°, 17.54±0.20°, 18.35±0.20°, 18.60±0.20°, 19.22±0.20°, 19.52±0.20°, 20.03±0.20°, 20.29±0.20°, 20.96±0.20°, 21.11±0.20°, 21.57±0.20°, 22.23±0.20°, 2 3.08±0.20°, 23.33±0.20°, 23.82±0.20°, 24.22±0.20°, 24.86±0.20°, 25.05±0.20°, 25.26±0.20°, 25.52±0.20°, 25.92±0.20°, 27.10±0.20°, 27.33±0.20°, 27.64±0.20°, 28.82±0.20°, 29.20± 0.20°, 29.68±0.20°, 30.21±0.20°, 30.59±0.20°, 31.19±0.20°, 31.75±0.20°, 32.52±0.20°, 33.71±0.20°, 34.51±0.20°, 35.88±0.20°, 36.54±0.20°, 37.45±0.20°, 38.23±0.20° and 38.60±0.20°.

6. The crystal according to claim 5, wherein the X-ray powder diffraction pattern has diffraction peaks at the following 2θ angles: 6.20°, 9.04°, 9.60°, 10.52°, 11.45°, 12.38°, 12.82°, 13.21°, 13.88°, 15.59°, 15.99, 16.59°, 17.18°, 17.54°, 18.35°, 18.60°, 19.22°, 19.52°, 20.03°, 20.29°, 20.96°, 21.11°, 21.57°, 2 2.23°, 23.08°, 23.33°, 23.82°, 24.22°, 24.86°, 25.05°, 25.26°, 25.52°, 25.92°, 27.10°, 27.33°, 27.64°, 28.82°, 29.20°, 29.68°, 30.21°, 30.59°, 31.19°, 31.75°, 32.52°, 33.71°, 34.51°, 35.88°, 36.54°, 37.45°, 38.23° and 38.60°.

7. The crystals of the compound of formula (I), whose XRPD pattern is substantially as shown in Figure 1. 8 . The crystal according to claim 1 , wherein the differential scanning calorimetry curve thereof has an endothermic peak starting point at 215.9° C.±5° C.

9. The crystal according to claim 8, wherein the DSC spectrum is substantially as shown in FIG2.

10. The crystal according to any one of claims 1 to 7, wherein the weight loss of the crystal at 200°C ± 3°C is 0.90%. The crystal according to claim 10 , wherein the TGA pattern is substantially as shown in FIG3 .

12. A crystalline composition according to any one of claims 1 to 11, wherein the crystals of the compound of formula (I) account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

13. A pharmaceutical composition comprising a therapeutically or prophylactically effective amount of the crystal according to any one of claims 1 to 11 or the crystalline composition according to claim 12.

14. Use of the crystal according to any one of claims 1 to 11, the crystalline composition according to claim 12, or the pharmaceutical composition according to claim 13 in the preparation of a medicament for a mineralocorticoid receptor antagonist-related disease; preferably, the mineralocorticoid receptor antagonist-related disease is selected from diabetic nephropathy.

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