Crystal form of p38alpha-MK2 inhibitor as well as preparation method and application of crystal form

Abstract

The invention provides an amorphous or polymorphic substance of a compound shown in a formula (I), and the amorphous or polymorphic substance has excellent stability and is suitable for industrial production and storage. # imgabs0 #

Description

[0001] This application claims the benefit of priority of the following prior patent applications:

[0002] The applicant's prior application, patent application number 202410210590.4, filed with the State Intellectual Property Office of China on February 26, 2024, entitled "A crystal form of a p38α-MK2 inhibitor, its preparation method, and application";

[0003] The entire contents of the above-mentioned prior patent applications are incorporated into this application by reference. Technical Field

[0004] The present invention belongs to the field of compounds, and particularly relates to a crystal form of a p38α-MK2 inhibitor, a preparation method and an application thereof. Background Art

[0005] PCT / CN2023 / 115579 (filing date August 29, 2023) describes a compound represented by formula (I). Pharmacological experiments show that the compound has good p38α-MK2 inhibitory effect and good selectivity for p38α-MK2. At the same time, the compound has good pharmacokinetic absorption and high bioavailability.

[0006]

[0007] The crystal structure of a pharmaceutically active ingredient often affects the chemical stability of the drug. Differences in crystallization and storage conditions can lead to variations in the compound's crystal structure, sometimes resulting in the formation of alternative crystalline forms. Therefore, in-depth research into the crystal forms of the compound represented by formula (I) and related preparation methods is essential to improve various properties of the compound represented by formula (I). Summary of the Invention

[0008] The present invention provides an amorphous or polymorphic compound of a compound represented by formula (I);

[0009]

[0010] According to an embodiment of the present invention, the amorphous form of the compound represented by formula (I) has substantially the following Figure 1 The X-ray powder diffraction pattern (XRPD pattern) is shown.

[0011] According to an embodiment of the present invention, the amorphous differential scanning calorimetry analysis spectrum (DSC diagram) of the compound represented by formula (I) is as follows Figure 2 shown.

[0012] According to an embodiment of the present invention, the polymorph of the compound represented by formula (I) is selected from the crystalline form A of the compound represented by formula (I).

[0013] According to an embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles:

[0014] 5.58°±0.20°,11.12°±0.20°,13.22°±0.20°,13.73°±0.20°,16.66°±0.20°.

[0015] According to an embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) further includes the following one, two or more characteristic diffraction peaks at 2θ angles:

[0016] 18.61°±0.20°,22.20°±0.20°,23.19°±0.20°,25.93°±0.20°,27.06°±0.20°.

[0017] According to an embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) further includes the following one, two or more characteristic diffraction peaks at 2θ angles:

[0018] The peak value of the ridge is 2.43°±0.20°, which is 7.66°±0.20°,9.26°±0.20°,9.67°±0.20°,16.00°±0.20°,17.36°±0.20°,18.02°±0.20°,19.25°±0.20°,28.99°±0.20°,30.15°±0.20°,30.64°±0.20°,30.97°±0.20°,31.42°±0.20°,32.91°±0.20°,33.49°±0.20°,34.06°±0.20°,37.03°±0.20°,38.74°±0.20°,41.72°±0.20°.

[0019] According to an embodiment of the present invention, the X-ray powder diffraction pattern analysis data of the crystalline form A of the compound represented by formula (I) is shown in Table 3, wherein the error range of 2θ of each characteristic diffraction peak is ±0.2°.

[0020] According to an embodiment of the present invention, the crystalline form A of the compound represented by formula (I) has substantially Figure 3 The X-ray powder diffraction pattern (XRPD pattern) is shown.

[0021] According to an embodiment of the present invention, the crystalline form A of the compound represented by formula (I) has almost no weight loss at room temperature to 100° C. According to an embodiment of the present invention, the crystalline form A is an anhydrous crystalline form.

[0022] According to an embodiment of the present invention, the thermogravimetric analysis spectrum (TGA spectrum) of the crystalline form A of the compound represented by formula (I) is as follows Figure 5 shown.

[0023] According to an embodiment of the present invention, the differential scanning calorimetry analysis spectrum (DSC spectrum) of the crystalline form A of the compound represented by formula (I) comprises an endothermic peak with a peak temperature of 163.5°C±2.0°C.

[0024] According to an embodiment of the present invention, the differential scanning calorimetry analysis spectrum (DSC diagram) of the crystalline form A of the compound represented by formula (I) is as follows Figure 6 shown.

[0025] The present invention also provides a method for preparing the amorphous or polymorphic form of the compound represented by the above formula (I), wherein:

[0026] The preparation method of the amorphous form of the compound represented by formula (I) is as follows: melting the compound represented by formula (I) and cooling to obtain the amorphous form of the compound represented by formula (I);

[0027] The preparation method of the polymorph of the compound represented by formula (I) is as follows: the compound represented by formula (I) or its amorphous form is dispersed or dissolved in a solvent, and crystallized to obtain the polymorph of the compound represented by formula (I).

[0028] According to an embodiment of the present invention, during the preparation of the amorphous form of the compound represented by formula (I), the melting temperature is 150°C-200°C, for example 170°C.

[0029] According to an embodiment of the present invention, during the preparation of the amorphous form of the compound represented by formula (I), the cooling temperature is 10° C.-30° C., such as room temperature.

[0030] According to an embodiment of the present invention, the preparation method of the crystalline form A of the compound represented by formula (I) is selected from the following method A or method B:

[0031] Method A: suspending the amorphous form of the compound represented by formula (I) in a first solvent and crystallizing to obtain the crystalline form A of the compound represented by formula (I);

[0032] Method B: dissolving the amorphous form of the compound represented by formula (I) in a second solvent, then adding a third solvent and crystallizing to obtain the crystalline form A of the compound represented by formula (I).

[0033] According to an embodiment of the present invention, the method for preparing the amorphous form of the compound represented by formula (I) is as described above.

[0034] According to an embodiment of the present invention, the method A is specifically as follows: suspending the amorphous form of the compound represented by formula (I) in a first solvent, crystallizing, centrifuging, and drying to obtain the crystalline form A of the compound represented by formula (I).

[0035] According to an embodiment of the present invention, in the method A, the first solvent is selected from any one, two or more of methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, dimethyl sulfoxide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, n-propanol, ethyl formate, butyl formate, toluene, dichloromethane, chloroform, water, cyclohexane, n-heptane, isopropyl ether, and methyl tert-butyl ether; in one embodiment, the first solvent is selected from any one, two or more of methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, dimethyl sulfoxide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, n-propanol , ethyl formate, butyl formate, toluene, dichloromethane, chloroform, any one, two or more thereof; in one embodiment, the first solvent is selected from any one, two or more thereof: water, cyclohexane, n-heptane, isopropyl ether, methyl tert-butyl ether; preferably, the first solvent is a mixed solvent consisting of solvent A and solvent B, solvent A is selected from any one of methanol, ethanol, isopropyl alcohol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, dimethyl sulfoxide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, n-propanol, ethyl formate, butyl formate, toluene, dichloromethane, chloroform, and solvent B is selected from any one of water, cyclohexane, n-heptane, isopropyl ether, methyl tert-butyl ether.

[0036] According to an exemplary embodiment of the present invention, in method A, the first solvent is selected from isopropanol, or a mixed solvent of acetonitrile and water (for example, the volume ratio of acetonitrile to water is 1:5-20, for example 1:8, 1:10, 1:12, 1:15).

[0037] According to an embodiment of the present invention, in the method A, the suspension temperature is 10°C-70°C, such as room temperature, 20°C, 30°C, 40°C, or 50°C.

[0038] According to an embodiment of the present invention, in the method A, the suspension time is 0.5 days to 7 days, for example, 1 day, 3 days, or 5 days.

[0039] According to an embodiment of the present invention, in the method A, the mass volume ratio of the amorphous form of the compound represented by formula (I) to the first solvent is 10 mg-100 mg:1 mL, for example, 90 mg:1 mL, 80 mg:1 mL, 70 mg:1 mL, 60 mg:1 mL, 50 mg:1 mL, 40 mg:1 mL, 30 mg:1 mL, and 20 mg:1 mL.

[0040] According to an embodiment of the present invention, in the method A, the drying condition is vacuum drying.

[0041] According to an embodiment of the present invention, in the method A, the drying temperature is 10°C-50°C, for example 40°C or room temperature.

[0042] According to an embodiment of the present invention, in the method A, the drying time is 2 hours to 24 hours, for example, overnight or 12 hours.

[0043] According to an embodiment of the present invention, the method B is specifically as follows: dissolving the amorphous form of the compound represented by formula (I) with a second solvent, then adding a third solvent, crystallizing, centrifuging, and drying to obtain the crystalline form A of the compound represented by formula (I).

[0044] According to an embodiment of the present invention, in the method B, the second solvent is selected from any one, two or more of dimethyl sulfoxide, methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, dichloromethane, and toluene.

[0045] According to an embodiment of the present invention, in the method B, the third solvent is selected from any one, two or more of water, cyclohexane, n-heptane and isopropyl ether.

[0046] According to an embodiment of the present invention, in the method B, the volume ratio of the second solvent to the third solvent is 1:1-20, for example, 1:3, 1:5, 1:8, 1:10, 1:12, 1:15, 1:18.

[0047] According to an embodiment of the present invention, in the method B, the mass volume ratio of the amorphous form of the compound represented by formula (I) to the second solvent is 1g:1mL-10mL, for example, 1g:1.5mL, 1g:2.0mL, 1g:2.5mL, 1g:3.0mL, 1g:5.0mL, 1g:8.0mL.

[0048] According to an embodiment of the present invention, in the method B, the mass volume ratio of the amorphous form of the compound represented by formula (I) to the third solvent is 1g:10mL-100mL, for example, 1g:15mL, 1g:20mL, 1g:25mL, 1g:30mL, 1g:50mL, 1g:80mL.

[0049] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the amorphous or polymorphic forms of the compound represented by the above formula (I).

[0050] According to an embodiment of the present invention, the pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients.

[0051] The excipients in the pharmaceutical composition are "pharmaceutically acceptable" in that they are compatible with the active ingredient of the composition (and preferably, capable of stabilizing the active ingredient) and are not deleterious to the subject being treated. One, two or more pharmaceutical excipients can be used to deliver the active compound.

[0052] According to an embodiment of the present invention, the pharmaceutical composition may further contain one, two or more additional therapeutic agents.

[0053] The present invention also provides the use of the amorphous or polymorphic form of the compound represented by the above formula (I), or the above pharmaceutical composition in the preparation of drugs.

[0054] According to an embodiment of the present invention, the drug is a drug for diagnosing, preventing and / or treating a disease or disorder mediated by p38α-MK2.

[0055] According to an embodiment of the present invention, the drug is a p38α-MK2 inhibitor.

[0056] According to an embodiment of the present invention, the p38α-MK2-mediated disease or symptom is selected from the group consisting of ulcerative colitis, inflammatory bowel inflammation, Crohn's disease, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, bone disease, osteoarthritis, septic shock, endotoxin shock, arthritis, sepsis, asthma, chronic obstructive pulmonary disease, cryopyrin-associated periodic syndrome, rheumatoid arthritis, hidradenitis suppurativa, ankylosing spondylitis or cancer.

[0057] According to an embodiment of the present invention, the amorphous or polymorphic form of the compound shown in the above formula (I) or the above pharmaceutical composition can be made into a form suitable for administration by any appropriate route, and one or more pharmaceutically acceptable carriers are used to prepare it by conventional methods. Therefore, the amorphous or polymorphic form of the compound shown in the above formula (I) or the above pharmaceutical composition can be formulated into various dosage forms for oral administration, injection (e.g., intravenous, intramuscular or subcutaneous) administration, inhalation or insufflation administration; It can also be formulated into sustained release dosage forms, such as tablets, hard or soft capsules, aqueous or oily suspensions, emulsions, injections, dispersible powders or granules, suppositories, lozenges or syrups.

[0058] The present invention also provides a method for diagnosing, preventing and / or treating a disease or condition mediated by p38α-MK2, comprising administering a therapeutically effective amount of an amorphous or polymorphic form of the compound represented by formula (I) or at least one of the pharmaceutical compositions described above to a patient in need of treatment.

[0059] According to an embodiment of the present invention, the p38α-MK2-mediated disease or symptom is selected from the group consisting of ulcerative colitis, inflammatory bowel inflammation, Crohn's disease, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, bone disease, osteoarthritis, septic shock, endotoxin shock, arthritis, sepsis, asthma, chronic obstructive pulmonary disease, cryopyrin-associated periodic syndrome, rheumatoid arthritis, hidradenitis suppurativa, ankylosing spondylitis or cancer.

[0060] According to an embodiment of the present invention, the patient is a mammal, preferably a human.

[0061] The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

[0062] The term "therapeutically effective amount" refers to that amount of an active compound or drug that will elicit the biological or medical response that a researcher, veterinarian, physician, or other clinician is seeking in a tissue, system, animal, individual, or human, and includes one or more of the following: (1) prevents disease, e.g., prevents a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who is not yet experiencing or developing the pathology or symptoms of the disease. (2) inhibits disease, e.g., inhibits the disease, disorder, or condition (i.e., prevents further development of the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition. (3) alleviates disease, e.g., alleviates the disease, disorder, or condition (i.e., reverses the pathology and / or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition.

[0063] Beneficial effects

[0064] The present invention provides an amorphous or polymorphic form of a compound represented by formula (I). The amorphous and polymorphic forms have excellent stability and are suitable for industrial production, manufacturing, and storage. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 : XRPD pattern of the amorphous form of the compound represented by formula (I);

[0066] Figure 2 : DSC spectrum of the amorphous form of the compound represented by formula (I);

[0067] Figure 3 : XRPD pattern of Form A of the compound represented by formula (I);

[0068] Figure 4 : NMR spectrum of Form A of the compound represented by formula (I);

[0069] Figure 5: TGA spectrum of Form A of the compound represented by formula (I);

[0070] Figure 6 : DSC spectrum of Form A of the compound represented by formula (I);

[0071] Figure 7 : DVS image of Form A of the compound represented by formula (I);

[0072] Figure 8 : XRPD comparison of Form A of the compound represented by formula (I) before and after DVS testing;

[0073] Figure 9 : XRPD comparison chart of stability study of Form A of the compound represented by formula (I);

[0074] Figure 10 : XRPD comparison chart of amorphous stability study of the compound represented by formula (I). DETAILED DESCRIPTION

[0075] The technical solutions of the present invention will be described in further detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are encompassed within the scope of protection that the present invention is intended to protect.

[0076] Patent application PCT / CN2023 / 115579 (filing date August 29, 2023) describes the compound represented by formula (I), and all contents involved in this patent application are added to the present invention by citation.

[0077] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0078] Unless otherwise stated, the following are the instruments, parameters, characterizations, and test methods used in the examples:

[0079] (1) NMR analysis 1 H NMR)

[0080] Several milligrams of solid sample were dissolved in dimethyl sulfoxide-d6 solvent and subjected to nuclear magnetic resonance analysis on a Bruker AVANCE NEO 400 (Bruker, Germany).

[0081] (2) X-ray powder diffraction (XRPD)

[0082] The solid samples obtained in the experiment were analyzed using a Bruker D8 Advance X-ray powder diffractometer (Bruker, Germany). The 2θ scan angle ranged from 3° to 45°, with a scan step size of 0.02° and an exposure time of 0.08 seconds. The test method used Cu target Kα1 radiation, a voltage of 40 kV, a current of 40 mA, and a zero-background sample pan.

[0083] (3) Thermogravimetric analysis (TGA)

[0084] The thermogravimetric analyzer (TA Discovery 550, US) was used. A 2-5 mg sample was placed in a equilibrated open aluminum sample pan and automatically weighed in the TGA furnace. The sample was heated to the final temperature at a rate of 10°C / min. A nitrogen purge rate of 60 mL / min was maintained at the sample and 40 mL / min at the balance.

[0085] (4) Differential Scanning Calorimetry (DSC)

[0086] The differential scanning calorimeter was a TA Discovery 250 (TA, US). 1-2 mg of sample was accurately weighed and placed in a perforated DSC Tzero sample pan. The sample was heated to the final temperature at a rate of 10°C / min, with nitrogen purge rate of 50 mL / min.

[0087] (5) Dynamic moisture adsorption and desorption analysis (DVS)

[0088] Dynamic moisture sorption / desorption analysis was performed using a DVS Intrinsic (SMS, UK). The test used a gradient mode with humidity changes from 0% to 95% to 0%, with each gradient increasing by 10% within the 0% to 90% range. The gradient endpoint was determined using the dm / dt method, with a dm / dt of less than 0.002% maintained for 10 minutes, or a maximum of 180 minutes per gradient. After the test, the samples were analyzed by XRPD to confirm any changes in the solid form.

[0089] (6) High performance liquid chromatography (HPLC)

[0090] The model of the HPLC reverse phase test was Shimadzu LC-2030C 3D plus (Shimadzu, JP), and the model of the HPLC normal phase test was ACQUITY ARC-2489 (Waters, US). The test conditions are shown in Tables 1 and 2.

[0091] Table 1 HPLC reverse phase test conditions

[0092]

[0093] Table 2 HPLC normal phase test conditions

[0094]

[0095] Example 1

[0096]

[0097] The first step is the preparation of 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

[0098] Under nitrogen protection, 1,4-dioxane (30 mL) was added to 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a (2.8 g, 6.8 mmol), tributyl(1-ethoxyvinyl)stannane (3.0 g, 8.2 mmol) and bis(tetrakistriphenylphosphine)palladium dichloride (0.48 g, 0.68 mmol). The reaction solution was stirred at 130 degrees Celsius for 12 hours. After the reaction solution was cooled to room temperature, potassium fluoride solution was added to quench the reaction and stirred for 30 minutes. The reaction mixture was stirred for 30 minutes, then filtered. The filtrate was extracted three times with ethyl acetate, concentrated, dissolved in tetrahydrofuran (20 mL), and concentrated hydrochloric acid (10 mL) was added. The mixture was stirred at room temperature for 30 minutes. The pH of the reaction mixture was then neutralized to 7 with 4N sodium hydroxide in an ice bath. The mixture was extracted three times with ethyl acetate. The mixture was dried, concentrated, and separated and purified on a silica gel column (petroleum ether / ethyl acetate = 35 / 65) to obtain 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001b (2.2 g) in a yield of 77%. MS m / z (ESI): 418.1 (M+1).

[0099] Step 2 Preparation of 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

[0100] To 2'-acetyl-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one 001b (1.5 g, 3.6 mmol) and NBS (0.64 g, 3.6 mmol) was added N,N-dimethylformamide (15 mL). The reaction solution was stirred at 60°C for 1 hour. After completion of the reaction, the mixture was dried, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 35 / 65) to afford 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one 001c (1.35 g) in a yield of 76%. MS m / z (ESI): 496.0 (M+1).

[0101] Step 3 Preparation of (E)-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0102] To 2'-acetyl-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001c (1.35 g, 2.7 mmol) and DMF-DMA (1.4 mL, 10.9 mmol) was added N,N-dimethylformamide (10 mL). The reaction mixture was heated at 55 °C. The mixture was stirred overnight. After the reaction, the mixture was directly dried and separated and purified on a silica gel column (dichloromethane / methanol = 95 / 5) to obtain (E)-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001d (0.91 g) in a yield of 67%. MS m / z (ESI): 551.0 (M+1). Step 4 Preparation of 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5'-6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0103] To (E)-3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-2'-(3-(dimethylamino)acryloyl)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001d (600 mg, 1.1 mmol), 2-hydroxy-2-methylpropionamidine (561 mg, 5.5 mmol) and potassium carbonate (607 mg, 4.4 mmol) was added N,N-dimethylformamide (4 mL). The reaction mixture was stirred at 4°C. The mixture was stirred at 55 degrees Celsius overnight. After the reaction, the mixture was washed with saturated brine and extracted with ethyl acetate. The organic phases were combined, dried, concentrated, and separated and purified on a silica gel column (ethyl acetate) to give a crude product, which was then purified on a preparative column to give 3-bromo-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-2'-(2-(2-hydroxypropane-2-yl)pyrimidin-4-yl)-5',6-dimethyl-2H-[1,4'-bipyridine]-2-one 001 (160 mg) in a yield of 25%.

[0104] Compound 001 was subjected to chiral separation to obtain an optically pure compound represented by formula (I) (t R =1.215min,30mg).

[0105] Split conditions:

[0106] Instrument brand: Waters Acquity UPCC;

[0107] Preparative column model: Daicel CHIRALPAK AD_3, 3*150mm, 3μm;

[0108] Mobile phase: CO2 / MeOH (0.1% DEA) = 80 / 20. Mass spectrometry, HPLC, 1 The H NMR data are as follows:

[0109] MS m / z(ESI):590.1(M+1).

[0110] HPLC: 91.24% (214nm), 90.34% (254nm).

[0111] 1H NMR (400MHz, DMSO-d6): δ9.01(d,J=5.2Hz,1H),8.76(s,1H),8.58(d,J=2.3Hz,1H),8.09–8.03(m,1H),8.02(d,J=5.2H z,1H),6.65(s,1H),6.06(q,J=6.3Hz,1H),5.05(s,1H),2.15(s,3H),1.98(s,3H),1.73(d,J=6.4Hz,3H),1.51(s,6H).

[0112] The compound represented by the above formula (I) was added to a glass bottle, placed in a blast drying oven, and then heated to 170°C. After the sample was completely melted, the glass bottle was left to cool at room temperature to obtain a light yellow solid. The solid was collected and characterized by XRPD. The spectrum is shown as follows: Figure 1 As shown, the results show that the light yellow solid obtained is an amorphous form. DSC analysis is performed on it, and its spectrum is as shown Figure 2 shown.

[0113] Preparation of intermediate 001a

[0114]

[0115] The first step is the preparation of 2-chloro-3-fluoro-5-methylpyridin-4-amine

[0116] 1,4-Dioxane (500 mL) was added to 2-chloro-3-fluoro-5-iodopyridin-4-amine 001a-1 (50 g, 0.18 mol), methylboronic acid (13.2 g, 0.22 mol), Pd(dppf)Cl2 (13.4 g, 0.018 mol), and cesium carbonate (119.6 g, 0.36 mol). The reaction mixture was stirred at 100°C for 16 hours. After completion of the reaction, the mixture was filtered, concentrated, and purified on a silica gel column (petroleum ether / ethyl acetate = 3 / 1) to afford 2-chloro-3-fluoro-5-methylpyridin-4-amine 001a-2 (17.4 g) in a 60% yield. MS m / z (ESI): 161.1 (M+1).

[0117] Step 2 Preparation of N-(2-chloro-3-fluoro-5-methylpyridin-4-yl)-2,6-dimethyl-4-oxo-4H-pyran-3-carboxamide

[0118] DMA (30 mL) was added to 2-chloro-3-fluoro-5-methylpyridin-4-amine 001a-2 (8.7 g, 0.054 mol) and 2,2,6-trimethyl-4H-1,3-dioxin-4-one (38.5 g, 0.27 mol). The reaction mixture was stirred at 115°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature, water (50 mL) was added, and the mixture was stirred. The reaction mixture was filtered to reveal N-(2-chloro-3-fluoro-5-methylpyridin-4-yl)-2,6-dimethyl-4-oxo-4H-pyran-3-carboxamide 001a-3, which was used directly in the next step. MS m / z (ESI): 311.1 (M+1).

[0119] Step 3 Preparation of 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0120] Water (50 mL) was added to the crude product from the previous step, followed by a 4M hydrochloric acid solution in dioxane (50 mL). The reaction mixture was stirred at 85°C for 16 hours. After completion of the reaction, the dioxane was partially removed by rotary evaporation and filtered to reveal the residue: 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a-4 (11.5 g). MS m / z (ESI): 311.1 (M+1).

[0121] Step 4 Preparation of 2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0122] Concentrated sulfuric acid (50 mL) was added to 3-acetyl-2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a-4 (11.5 g, 0.037 mol). The reaction solution was stirred at 100 degrees Celsius for 16 hours. After the reaction was completed, it was cooled to room temperature and added to ice water. The pH was adjusted to 9-10 with NaOH and then to 3-4 with concentrated hydrochloric acid. The reaction solution was filtered to obtain the residue 2'-chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a-5 (13 g). MS m / z (ESI): 269.0 (M+1). Step 5 Preparation of 1-(3,5-difluoropyridin-2-yl)ethan-1-ol

[0123] 1-(3,5-Difluoropyridin-2-yl)ethan-1-one 001a-A (1.0 g, 6.4 mmol) was dissolved in methanol (10 mL), followed by the slow addition of sodium borohydride (0.06 g, 1.6 mmol). The reaction mixture was stirred at -10°C for 0.5 hours. After completion of the reaction, the reaction solution was washed with saturated ammonium chloride and extracted with ethyl acetate. The organic phase was then dried, filtered, concentrated, and purified by silica gel column chromatography (mobile phase: ethyl acetate / dichloromethane = 20%-50%) to afford 1-(3,5-difluoropyridin-2-yl)ethan-1-ol 001a-B (0.7 g) in a 65% yield. MS m / z (ESI): 160.1 (M+1).

[0124] Step 6 Preparation of 2-(1-chloroethyl)-3,5-difluoropyridine

[0125] 1-(3,5-Difluoropyridin-2-yl)ethan-1-ol 001a-B (500 mg, 3.14 mmol) was dissolved in dichloromethane (20 mL). Thionyl chloride (1121 mg, 9.43 mmol) was added dropwise at 0°C. The reaction was stirred at 50°C for 2 hours. After completion of the reaction, the reaction solution was concentrated to yield crude 2-(1-chloroethyl)-3,5-difluoropyridine 001a-C (500 mg). Yield: 71.69%. MS m / z (ESI): 178.0 (M+1).

[0126] Step 7 Preparation of 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0127] 2'-Chloro-3'-fluoro-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a-5 (4 g, 14.9 mmol), 2-(1-chloroethyl)-3,5-difluoropyridine 001a-C (3.97 g, 22.3 mmol), 18-crown-6 (390 mg, 1.4 mmol), and potassium carbonate (6.18 g, 44.7 mmol) were dissolved in N,N-dimethylformamide (15 mL) and stirred at 60°C for 12 hours. After the reaction, the mixture was washed with water (20 mL) and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined and washed with saturated brine (20 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (ethyl acetate / petroleum ether = 75%) to give 2'-chloro-4-(1-(3,5-difluoropyridin-2-yl)ethoxy)-3'-fluoro-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 001a (3.7 g) in a 54% yield. MS m / z (ESI): 410.1 (M+1).

[0128] Example 2

[0129] 600.1 mg of the amorphous compound of formula (I) in Example 1 was weighed, 10.0 mL of isopropanol was added thereto, and the mixture was suspended and stirred at room temperature for 3 days. The suspension was filtered, the filter cake was collected, and the solid was dried under vacuum at 40°C overnight to obtain the compound of formula (I) as a light yellow solid (335.2 mg, yield: 55.86%). XRPD characterization was performed on the compound, and the XRPD spectrum was as shown below. Figure 3 The characteristic peak positions are shown in Table 3, and the obtained light yellow solid product is Form A.

[0130] Table 3 Characteristic peaks of XRPD spectrum of Form A of the compound represented by formula (I)

[0131]

[0132]

[0133] The obtained crystal form A of the compound represented by formula (I) was subjected to NMR, TGA, DSC and DVS detection, and the spectrum was as follows: Figure 4-8 As shown. TGA results show that the Form A sample does not lose weight significantly when heated to 100°C, but may decompose above 260°C ( Figure 5 The DSC results showed that the crystal form A sample had a melting endothermic peak at 163.5℃ ( Figure 6 ). Thus, it was confirmed that Form A was an anhydrous crystalline form.

[0134] The DVS results showed that the Form A sample gained 0.26% weight at 80% RH, gained 0.36% weight at 95% RH, and lost 0.07% weight at 0% RH, indicating that the sample was slightly hygroscopic ( Figure 7 ); There was no significant change in the XRPD results of the samples before and after the DVS experiment ( Figure 8 ).

[0135] Example 3

[0136] Weigh 20.3 mg of the amorphous form of the compound represented by formula (I), add 0.1 mL of acetonitrile and 1 mL of water, and suspend and stir at 50°C for 1 day. The suspension is centrifuged and the solid is dried under vacuum at room temperature. The solid is characterized by XRPD. The XRPD spectrum is the same as that in Example 2. Figure 3 The results are basically consistent, indicating that the obtained crystal is form A.

[0137] Example 4

[0138] Weigh 20.1 mg of the amorphous form of the compound represented by formula (I), add 0.05 mL of dimethyl sulfoxide to dissolve it, and add the solution dropwise to 0.5 mL of water. The suspension is centrifuged and dried under vacuum at room temperature. The solid is characterized by XRPD. The XRPD spectrum is the same as that in Example 2. Figure 3 The results are basically consistent, indicating that the obtained crystal is form A.

[0139] Example 5 Biological Activity Test

[0140] 5.1 Experimental Objective: Determine the IC50 of the compounds for p38α-MK2 kinase inhibition 50 value.

[0141] 5.2 Experimental Materials:

[0142] GST-MK2 solution concentration: 15328.84nM

[0143] GST-P-p38α solution concentration: 3030nM

[0144] Hsp27 polypeptide (FITC) concentration: 1051.3 μM.

[0145] 5.3 Experimental operation:

[0146] (1) Use Echo to dilute the compound to a final concentration of 10 μM to 0.17 nM, and add 200 nL of the solution to the wells of the assay plate;

[0147] (2) Add 5 μL of 4-fold diluted GST-MK2 mixture to the wells of the assay plate (containing compound);

[0148] (3) Add 15 μL of a 1.33-fold diluted mixture of GST-P-p38a, ATP, and Hsp27 peptide to the wells of the assay plate (containing the compound);

[0149] (4) Centrifuge at 1000 rpm for about 60 seconds and incubate at 23°C for 120 minutes;

[0150] (5) Add 60 μL of IMAP solution to start the reaction;

[0151] (6) Centrifuge the plate at 1000 rpm for approximately 60 seconds and incubate at 23°C for 30 minutes;

[0152] (7) Read the detection plate on Neo2 (Ex / Em=485nm / FITC FP-P pol 528nm & FITC FP-S pol 528nm);

[0153] (8) Calculate the relative enzyme activity inhibition relative to the DMSO blank by signal ratio, and calculate the IC using software curve fitting.50 value.

[0154] 5.4 Experimental results: IC inhibitory activity of the compound represented by formula (I) on p38α-MK2 kinase 50 The value is 1.8nM.

[0155] Example 6 Stability Test

[0156] The stability of the crystalline form A and the amorphous form of the compound represented by formula (I) was studied under high temperature (60°C), high humidity (25°C / 92.5% RH), light (25°C / 4500 Lux), and accelerated (40°C / 75% RH) conditions. Samples were taken for XRPD characterization and HPLC characterization at 7 days and 15 days, respectively (the purity of the relevant substances, T0 purity, was tested using the HPLC reverse phase test conditions in Table 1; the normal phase main peak area was tested using the HPLC normal phase test conditions in Table 2). The results are shown in Tables 4 and 5. Figure 9-10 shown.

[0157] XRPD comparison results showed that Form A and amorphous form did not undergo crystal transformation under the four stability test conditions for 7 days and 15 days. HPLC test was performed on the samples after the stability test, and the HPLC results showed:

[0158] Compared with the T0 results, the chemical purity of Form A did not change significantly after 7 days and 15 days of stability testing under the four conditions, and the main peak area value of normal phase HPLC test did not change significantly;

[0159] Compared with the T0 results, the chemical purity of the amorphous form decreased significantly after 15 days of photostability experiment, and the main peak area value of the normal phase HPLC test also decreased significantly; after 15 days of high humidity stability experiment, the chemical purity of the amorphous form decreased, and the decrease value was smaller than the decrease value after the photostability experiment, and the main peak area value of the normal phase HPLC test did not decrease significantly; under the other experimental conditions, the chemical purity and the main peak area value of the normal phase HPLC test did not decrease significantly after the 15-day stability experiment.

[0160] Table 4 Stability study results

[0161]

[0162] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. an amorphous or polymorphic form of the compound represented by formula (I), 2. The amorphous or polymorphic compound according to claim 1, wherein The amorphous form of the compound represented by formula (I) has an X-ray powder diffraction pattern substantially as shown in FIG1 ; Preferably, the differential scanning calorimetry analysis spectrum of the amorphous form of the compound represented by formula (I) is shown in FIG2 .

3. The amorphous or polymorphic compound according to claim 1, wherein The polymorph of the compound represented by formula (I) is selected from the crystalline form A of the compound represented by formula (I).

4. The amorphous or polymorphic compound according to claim 3, wherein The X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 5.58°±0.20°,11.12°±0.20°,13.22°±0.20°,13.73°±0.20°,16.66°±0.20°； Preferably, the X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) further includes one, two or more of the following characteristic diffraction peaks at 2θ angles: 18.61°±0.20°,22.20°±0.20°,23.19°±0.20°,25.93°±0.20°,27.06°±0.20°； Preferably, the X-ray powder diffraction pattern of the crystalline form A of the compound represented by formula (I) further includes one, two or more of the following characteristic diffraction peaks at 2θ angles: 7.66°±0.20°,9.26°±0.20°,9.67°±0.20°,16.00°±0.20°，17.36°±0.20°，18.02°±0.20°，19.25°±0.20°，28.99°±0.20°，30.15°±0.20°，30.64°±0.20°，30.97°±0.20°，31.42°±0.20°，32.91°±0.20°，33.49°±0.20°，34.06°±0.20°，37.03°±0.20°，38.74°±0.20°，41.72°±0.20°； Preferably, the X-ray powder diffraction pattern analysis data of the crystalline form A of the compound represented by formula (I) is shown in Table 3, wherein the error range of 2θ of each characteristic diffraction peak is ±0.2°; Preferably, the crystalline form A of the compound represented by formula (I) has an X-ray powder diffraction pattern substantially as shown in FIG3 ; Preferably, the crystalline form A of the compound represented by formula (I) has almost no weight loss at room temperature to 100°C; Preferably, the thermogravimetric analysis spectrum of Form A of the compound represented by formula (I) is shown in FIG5 ; Preferably, the differential scanning calorimetry analysis spectrum of the crystalline form A of the compound represented by formula (I) comprises an endothermic peak with a peak temperature of 163.5°C±2.0°C; Preferably, the differential scanning calorimetry analysis spectrum of Form A of the compound represented by formula (I) is shown in FIG6 .

5. A method for preparing an amorphous or polymorphic form of the compound of formula (I) according to any one of claims 1 to 4, characterized in that: The preparation method of the amorphous form of the compound represented by formula (I) is as follows: melting the compound represented by formula (I) and cooling to obtain the amorphous form of the compound represented by formula (I); The preparation method of the polymorph of the compound represented by formula (I) is as follows: the compound represented by formula (I) or its amorphous form is dispersed or dissolved in a solvent, and crystallized to obtain the polymorph of the compound represented by formula (I).

6. The preparation method according to claim 5, characterized in that During the preparation of the amorphous form of the compound represented by formula (I), the melting temperature is 150°C-200°C; Preferably, during the preparation of the amorphous form of the compound represented by formula (I), the cooling temperature is 10°C-30°C.

7. The preparation method according to claim 5, characterized in that The preparation method of the crystalline form A of the compound represented by formula (I) is selected from the following method A or method B: Method A: suspending the amorphous form of the compound represented by formula (I) in a first solvent and crystallizing to obtain the crystalline form A of the compound represented by formula (I); Method B: dissolving the amorphous form of the compound represented by formula (I) in a second solvent, then adding a third solvent and crystallizing to obtain the crystalline form A of the compound represented by formula (I).

8. The preparation method according to claim 7, characterized in that The first solvent is selected from any one, two or more of methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, dimethyl sulfoxide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, n-propanol, ethyl formate, butyl formate, toluene, dichloromethane, chloroform, water, cyclohexane, n-heptane, isopropyl ether, and methyl tert-butyl ether; and / or, the second solvent is selected from any one, two or more of dimethyl sulfoxide, methanol, ethanol, isopropanol, acetone, tetrahydrofuran, dioxane, ethylene glycol methyl ether, ethylene glycol dimethyl ether, acetonitrile, dimethylformamide, isopropyl acetate, ethyl acetate, 4-methyl-2-pentanone, dichloromethane, and toluene; And / or, the third solvent is selected from any one, two or more of water, cyclohexane, n-heptane, and isopropyl ether.

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the amorphous form or polymorphic form of the compound of formula (I) according to any one of claims 1 to 4; Preferably, the pharmaceutical composition further comprises one, two or more pharmaceutically acceptable excipients; Preferably, the pharmaceutical composition may further contain one, two or more additional therapeutic agents.

10. Use of the amorphous or polymorphic form of the compound of formula (I) according to any one of claims 1 to 4, or the pharmaceutical composition according to claim 9 in the preparation of a medicament; Preferably, the drug is a drug for diagnosing, preventing and / or treating a disease or condition mediated by p38α-MK2; Preferably, the drug is a p38α-MK2 inhibitor; Preferably, the p38α-MK2-mediated disease or condition is selected from ulcerative colitis, inflammatory bowel disease, Crohn's disease, psoriasis, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, bone disease, osteoarthritis, septic shock, endotoxic shock, arthritis, sepsis, asthma, chronic obstructive pulmonary disease, cryopyrin-associated periodic syndrome, rheumatoid arthritis, hidradenitis suppurativa, ankylosing spondylitis or cancer.

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