Crystalline forms of 2-methyl-2-propanol and amino-substituted aryl compounds

Abstract

A crystalline form of 2-methyl-2-propanol and an amino-substituted aryl compound (Formula I) and a method for preparing the same, including the use of the crystalline form in preparing a drug for treating related diseases. #imgabs0#

Description

[0001] This application claims priority to:

[0002] CN202010554402.1, application date: 2020-06-17. Technical Field

[0003] The present invention relates to a crystal form of 2-methyl-2-propanol and an amino-substituted aryl compound and a preparation method thereof, and also relates to application of the crystal form in preparing medicines for treating related diseases. Background Art

[0004] Dry eye, also known as keratoconjunctivitis sicca, refers to a broad range of conditions characterized by abnormal tear quality, quantity, or dynamics, resulting in decreased tear film stability and accompanied by ocular discomfort or ocular surface pathology. Specific symptoms include eye irritation, visual impairment, and tear film instability. Some cases of this syndrome are caused by ocular surface inflammation, leading to lacrimal gland dysfunction. Systemic autoimmunity is also associated with this condition.

[0005] Because some toxic aldehydes are produced by the body or eye tissues and organs through metabolic mechanisms, such as malondialdehyde (MDA) and 4-hydroxy-2-nonenal (4HNE), these aldehydes are highly reactive with proteins, carbohydrates, oils and DNA, leading to chemical modification of biological molecules and activation of inflammatory molecule regulators such as NF-kappaB, thereby causing damage to different organs. This is one of the causes of dry eye.

[0006] Through research, the present invention has found that small molecule drugs can be taken as eye drops or orally to enter the inflamed area of the eye, and through complexation reaction with aldehydes in the body, the toxicity of aldehydes is reduced, inflammation is reduced, and the effect of treating dry eye is achieved. Summary of the Invention

[0007] The present invention provides a crystal form A of a compound of formula (I), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 16.36±0.20°, and 21.23±0.20°.

[0008]

[0009] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 8.20±0.20°, 16.36±0.20°, 17.73±0.20°, 20.69±0.20°, 21.23±0.20°, 22.51±0.20°, 28.36±0.20°.

[0010] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 8.20±0.20°, 14.16±0.20°, 16.36±0.20°, 17.73±0.20°, 19.42±0.20°, 20.69±0.20°, 21.23±0.20°, 22.51±0.20°, 28.36±0.20°.

[0011] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 7.03°, 8.20°, 9.19°, 12.53°, 13.09°, 14.16°, 15.78°, 16.36°, 17.73°, 18.46°, 19.42°, 20.15°, 20.69°, 21.23°, 22.51°, 23.45°, 24.02°, 24.86°, 27.36°, 28.36°, 31.36°, 32.41°, and 38.61°.

[0012] In some embodiments of the present invention, the above-mentioned crystal form A has an XRPD pattern as shown in FIG. Figure 1 shown.

[0013] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form A are shown in Table 1:

[0014] Table 1 XRPD pattern analysis data of the crystal form of compound A of formula (I)

[0015]

[0016] The present invention provides a crystal form B of a compound of formula (I), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 14.28±0.20°, 14.78±0.20°, and 20.18±0.20°.

[0017]

[0018] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 10.08±0.20°, 12.60±0.20°, 14.28±0.20°, 14.78±0.20°, 20.18±0.20°, 21.32±0.20°, 26.08±0.20°, and 27.69±0.20°.

[0019] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 10.08±0.20°, 12.60±0.20°, 14.28±0.20°, 14.78±0.20°, 20.18±0.20°, 21.32±0.20°, 25.26±0.20°, 26.08±0.20°, 27.69±0.20°, and 29.63±0.20°.

[0020] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 8.09°, 10.08°, 12.60°, 14.28°, 14.78°, 15.13°, 16.08°, 16.38°, 17.51°, 18.25°, 20.18°, 20.56°, 21.12°, 21.32°, 22.65°, 2 3.85°, 24.15°, 24.77°, 25.26°, 26.08°, 26.34°, 26.64°, 27.69°, 28.94°, 29.12°, 29.63°, 30.43°, 32.16°, 32.53°, 33.34°, 33.74°, 34.74°, 35.13°, 36.20°, 37.95°, 38.24°.

[0021] In some embodiments of the present invention, the above-mentioned B crystal form has an XRPD pattern as shown in FIG. Figure 2 shown.

[0022] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form B are shown in Table 2:

[0023] Table 2 XRPD pattern analysis data of the crystal form of compound B of formula (I)

[0024]

[0025] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned B crystal form has an endothermic peak at 183.85±3.0°C.

[0026] In some embodiments of the present invention, the DSC spectrum of the above-mentioned B crystal form is as follows Figure 7 shown.

[0027] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned Form B shows a weight loss of 0.339% at 172.58°C±3.0°C.

[0028] In some embodiments of the present invention, the TGA spectrum of the above-mentioned B crystal form is as follows Figure 7 shown.

[0029] The present invention provides a crystal form C of a compound of formula (I), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 11.91±0.20°, 18.36±0.20°, and 19.46±0.20°.

[0030]

[0031] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.97±0.20°, 10.66±0.20°, 11.91±0.20°, 16.09±0.20°, 18.36±0.20°, 19.46±0.20°, 19.89±0.20°, 23.99±0.20°.

[0032] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.97±0.20°, 10.66±0.20°, 11.91±0.20°, 12.65±0.20°, 16.09±0.20°, 18.36±0.20°, 19.46±0.20°, 19.89±0.20°, 23.99±0.20°, 25.47±0.20°.

[0033] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.97°, 10.66°, 11.91°, 12.65°, 14.06°, 14.82°, 15.73°, 16.09°, 17.82°, 18.36°, 19.46°, 19.89°, 20.16°, 20.82°, 21.35 °, 21.93°, 23.99°, 25.47°, 26.42°, 27.33°, 28.24°, 29.26°, 31.26°, 31.96°, 32.31°, 32.69°, 33.19°, 33.79°, 34.93°, 35.48°, 36.05°, 36.74°, 37.78°, 38.57°, 39.32°.

[0034] In some embodiments of the present invention, the above-mentioned C crystal form has an XRPD pattern as shown in FIG. Figure 3 shown.

[0035] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form C are shown in Table 3:

[0036] Table 3 XRPD pattern analysis data of the crystal form C of the compound of formula (I)

[0037]

[0038] The present invention provides a D crystal form of a compound of formula (II), characterized in that its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.020±0.200°, 16.243±0.200°, and 17.690±0.200°.

[0039]

[0040] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned D crystal form has characteristic diffraction peaks at the following 2θ angles: 7.020±0.200°, 8.131±0.200°, 16.243±0.200°, 17.690±0.200°, and 21.081±0.200°.

[0041] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned D crystal form has characteristic diffraction peaks at the following 2θ angles: 7.020±0.200°, 8.131±0.200°, 14.029±0.200°, 16.243±0.200°, 17.690±0.200°, 21.081±0.200°, and 22.286±0.200°.

[0042] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned D crystal form has characteristic diffraction peaks at the following 2θ angles: 7.020±0.200°, 8.131±0.200°, 12.461±0.200°, 14.029±0.200°, 16.243±0.200°, 17.690±0.200°, 20.549±0.200°, 21.081±0.200°, and 22.286±0.200°.

[0043] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned D crystal form has characteristic diffraction peaks at the following 2θ angles: 7.020°, 7.840°, 8.131°, 9.190°, 12.461°, 13.031°, 14.029°, 15.669°, 16.243°, 16.977°, 17.690°, 18.401°, 19.279°, 20.549°, 21.081°, 22.286°, 23.364°, 23.918°, 24.716°, 27.126°, and 28.209°.

[0044] In some embodiments of the present invention, the above-mentioned D crystal form has an XRPD pattern as shown in FIG. Figure 8 shown.

[0045] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned D crystal form are shown in Table 4:

[0046] Table 4 XRPD pattern analysis data of the crystal form of compound D of formula (II)

[0047]

[0048] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned D crystal form has an endothermic peak at 99.8±3.0°C and 181.97.0±3.0°C, respectively.

[0049] In some embodiments of the present invention, the DSC spectrum of the above-mentioned D crystal form is as follows Figure 9 shown.

[0050] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned D crystal form shows a weight loss of 5.029% at 120.0°C±3.0°C.

[0051] In some embodiments of the present invention, the TGA spectrum of the above-mentioned D crystal form is as follows Figure 9 shown.

[0052] The present invention also provides the use of the above-mentioned crystal form A, crystal form B and crystal form C in the preparation of drugs for treating diseases associated with aldehyde binders.

[0053] The present invention also provides the use of the above-mentioned crystal form A, crystal form B, crystal form C and crystal form D in the preparation of medicines for treating diseases related to aldehyde scavengers.

[0054] In some embodiments of the present invention, the above application is characterized in that the drug for the aldehyde scavenger-related disease is a drug for dry eye.

[0055] Definition and Description

[0056] 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.

[0057] The intermediate compounds 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, 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 invention.

[0058] The chemical reactions of the present invention are carried out in suitable solvents that are compatible with the chemical transformations of the present invention and the reagents and materials required. To obtain the compounds of the present invention, it may sometimes be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0059] The present invention will be described in detail below through examples, which are not intended to limit the present invention in any way.

[0060] All solvents used in the present invention were commercially available and used without further purification.

[0061] The solvents used in the present invention are commercially available. The present invention uses the following abbreviations: DCM represents dichloromethane; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOH represents ethanol; MeOH represents methanol; TFA represents trifluoroacetic acid; ATP represents adenosine triphosphate; HEPES represents 4-hydroxyethylpiperazineethanesulfonic acid; and MgCl2 represents magnesium dichloride.

[0062] Technical Effects

[0063] The compound of the present invention has good crystal stability and is easy to formulate into medicine; the compound of the present invention has excellent aldehyde complexing ability, is highly safe, and has a low possibility of drug interaction; the compound of the present invention has good pharmacokinetic characteristics, helps to alleviate ocular inflammation, and achieves the purpose of treating dry eye.

[0064] X-ray powder diffractometer (XRPD) method of the present invention

[0065] Instrument model: PANalytical X'pert3 X-ray diffractometer

[0066] Test method: Approximately 10 mg of sample was used for XRPD analysis.

[0067] The detailed XRPD parameters are as follows:

[0068] Radiation source: Cu, Kα 1.540598, 1.544426; Kα2 / Kα1 intensity ratio: 0.50)

[0069] Voltage: 45 kilovolts (kV)

[0070] Current: 40 milliamperes (mA)

[0071] Divergence slit: 1 / 16 degree

[0072] Scan mode: continuous

[0073] Scanning range: from 3.0 to 40.0 degrees

[0074] Scan time per step: 46.665 seconds

[0075] Step size: 0.0263 degrees BRIEF DESCRIPTION OF THE DRAWINGS

[0076] Figure 1 is the XRPD spectrum of the crystal form A of the compound of formula (I) using Cu-Kα radiation;

[0077] Figure 2 is the XRPD spectrum of the crystal form B of the compound of formula (I) using Cu-Kα radiation;

[0078] Figure 3 is the XRPD spectrum of the crystal form C of the compound of formula (I) using Cu-Kα radiation;

[0079] Figure 4 This is the result of in vitro aldehyde capture ability test;

[0080] Figure 5 This is the tear secretion result of the dry eye efficacy experiment in mice;

[0081] Figure 6 This is the result of corneal fluorescence staining in the drug efficacy experiment for dry eye in mice;

[0082] Figure 7 The DSC and TGA spectra of the crystalline form B of the compound of formula (I) are shown below:

[0083] Figure 8 is the XRPD spectrum of the crystal form D of the compound of formula (II) using Cu-Kα radiation;

[0084] Figure 9 The DSC and TGA spectra of the crystal form D of compound of formula (II) are shown. DETAILED DESCRIPTION

[0085] In order to better understand the content of the present invention, further description is given below in conjunction with specific embodiments, but the specific implementation methods are not intended to limit the content of the present invention.

[0086] Example 1: Preparation of compound of formula (I)

[0087]

[0088] Synthesis route:

[0089]

[0090] Step 1: Synthesis of compound 1-2

[0091] To a solution of compound 1-1 (2 g, 7.19 mmol, 1 eq) in toluene (30 mL) were added bispinacol borate (3.65 g, 14.39 mmol, 2 eq), Pd(dppf)Cl2 (526.35 mg, 719.34 μmol, 0.1 eq), and potassium acetate (1.41 g, 14.39 mmol, 2 eq). The mixture was reacted at 110°C for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain crude product 1-2.

[0092] Step 2: Synthesis of Compounds 1-4

[0093] Compound 1-3 (2.3 g, 7.07 mmol, 1 eq), cesium carbonate (4.61 g, 14.14 mmol, 2 eq), and Pd(dppf)Cl2 (517.32 mg, 707.00 μmol, 0.1 eq) were added to a solution of compound 1-2 (1.63 g, 7.07 mmol, 1 eq) in toluene (50 mL) and water (10 mL). The mixture was reacted at 110°C under nitrogen for 6 hours. After the reaction was completed, the reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (SiO2, petroleum ether / ethyl acetate = 5:1 to 0:1). Compound 1-4 was obtained. LCMS: 350.1 [M+1] + Step 3: Synthesis of Compounds 1-5

[0094] To a solution of compound 1-4 (1.58 g, 4.52 mmol, 1 eq) in methanol (50 mL) and ethyl acetate (50 mL) was added Pd / C (1.7 g, 5% purity). The mixture was reacted under a hydrogen atmosphere at 15 psi and 20°C for 3 hours. After completion of the reaction, the reaction solution was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain compound 1-5. LCMS: 320 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm7.96 (t, J=1.76Hz, 1H), 7.54 (d, J=11.84Hz, 1H), 7.39 (s, 1H), 6.95 (d, J = 6.58Hz, 1H), 6.82 (s, 2H), 6.66 (s, 2H), 3.83 (d, J = 3.96Hz, 6H).

[0095] Step 4: Synthesis of compound of formula (I)

[0096] Methylmagnesium bromide (3M, 31.32 mL, 20 eq) was added to a solution of compound 1-5 (1.5 g, 4.70 mmol, 1 eq) in tetrahydrofuran (150 mL) at 0°C and allowed to react at 20°C for 3 hours. After completion of the reaction, the reaction solution was poured into a saturated ammonium chloride solution and separated by adding 50 mL of ethyl acetate. The aqueous phase was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by HPLC (column model: Xtimate C18 150 x 25 mm x 5 μm; mobile phase: [water (10 mM ammonium bicarbonate solution) - acetonitrile]; B (acetonitrile) %: 25%-45%, 10.5 min). Compound (I) was obtained. LCMS: 320.0 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm 7.77 (s, 1H), 7.05 (s, 1H), 6.91 (d, J=12.28Hz, 1H), 6.67 (d, J=7.46Hz, 1H), 5.63 (s, 2H), 5.27-5.48 (m, 4H), 1.51 (s, 12H)

[0097] In some embodiments of the present invention, the compound of formula (I) can also be prepared by the following route:

[0098]

[0099] Step 1: Preparation of compound 2

[0100] Compound 1 (100 g, 460.79 mmol, 1 eq) was dissolved in anhydrous ethanol (1 L), and concentrated sulfuric acid (225.97 g, 2.30 mol, 122.81 mL, 5 eq) and anhydrous sodium sulfate Na2SO4 (65.45 g, 460.79 mmol, 46.75 mL, 1 eq) were added. The reaction mixture was stirred at 85°C for 48 hours. After the reaction was completed, the reaction mixture was cooled to room temperature. Saturated sodium bicarbonate aqueous solution (1 L) was added dropwise to the reaction mixture. A large amount of solid was formed, which was filtered and the filter cake was washed with water (500 mL). The resulting solid was vacuum dried to obtain compound 2.

[0101] 1 H NMR (400MHz, CDCl3) δ 8.10 (d, J=1.8Hz, 1H), 7.26 (s, 1H), 4.47 (q, J=7.1Hz, 2H), 1.46 (t, J=7.2Hz, 3H).

[0102] Step 2: Preparation of compound 3

[0103] Compound 2 (70.00 g, 285.63 mmol, 1 eq) was dissolved in tetrahydrofuran (1 L) and cooled to -78°C under nitrogen. Methyllithium (1.6 M, 892.59 mL, 5 eq) was slowly added dropwise to the reaction solution, and the reaction solution was stirred at -78°C for 3 hours. After the reaction was completed, water (100 mL) was slowly added dropwise to quench the reaction. The solution was warmed to room temperature and diluted with saturated aqueous ammonium chloride (500 mL). The solution was extracted with ethyl acetate (500 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was slurried with n-heptane (500 mL), filtered, and dried to obtain compound 3.

[0104] 1 H NMR (400MHz, CDCl3) δ7.86 (d, J=1.9Hz, 1H), 6.98 (d, J=1.9Hz, 1H), 4.57 (brs, 2H), 1.57 (s, 6H).

[0105] Step 3: Preparation of compound 5

[0106] Compound 4 (50 g, 189.39 mmol, 1 eq) was dissolved in ethanol (500 mL). Concentrated sulfuric acid (92.88 g, 946.95 mmol, 50.48 mL, 5 eq) and anhydrous sodium sulfate (26.90 g, 189.39 mmol, 1 eq) were added, and the reaction mixture was stirred at 85°C for 72 hours. After completion of the reaction, the reaction mixture was slowly poured into a 10°C aqueous sodium carbonate solution (200 g of sodium carbonate dissolved in 5 liters of water), stirred at this temperature for 1 hour, filtered, and the filter cake washed with water (1 L). The resulting solid was vacuum-dried (15°C) to yield compound 5.

[0107] 1 H NMR (400MHz, CDCl3) δ8.22 (d, J=6.0Hz, 1H), 7.49 (d, J=7.5Hz, 1H), 4.43 (q, J=7.4Hz, 2H), 1.39 (t, J=7.3Hz, 3H).

[0108] Step 4: Preparation of compound 6

[0109] Compound 5 (20 g, 68.48 mmol, 1 eq), bis-naphthalene borate (34.78 g, 136.96 mmol, 2 eq), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane adduct (5.59 g, 6.85 mmol, 0.1 eq) and potassium acetate (13.44 g, 136.95 mmol, 2 eq) were added to dioxane (250 mL), the atmosphere was purged with nitrogen three times, and the reaction solution was stirred at 90 ° C for 4 hours. After completion of the reaction, the reaction solution was filtered through a pad of celite, the filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate = 10:1 to 2:1) to obtain a crude compound. This crude product was slurried with n-heptane (100 mL), stirred at 15 ° C for 12 hours, filtered, and the filter cake was dried to obtain compound 6.

[0110] 1 H NMR (400MHz, CDCl3) δ8.39 (d, J=5.0Hz, 1H), 7.35 (d, J=8.0Hz, 1H), 4.43 (q, J=7.0Hz, 2H), 1.42-1.38 (m, 15H).

[0111] Step 5: Preparation of compound 7

[0112] Compound 6 (15 g, 44.23 mmol, 1 eq), compound 3 (11.25 g, 48.68 mmol, 1.1 eq), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane adduct (3.63 g, 4.45 mmol, 0.1 eq), and cesium carbonate (43.23 g, 132.70 mmol, 3 eq) were added to dioxane (50 mL) and water (30 mL). The atmosphere was purged with nitrogen three times, and the reaction solution was stirred at 90°C for 12 hours. After completion of the reaction, the reaction solution was diluted with ethyl acetate (500 mL) and filtered through celite. The filtrate was washed with saturated brine (200 mL), and the organic phase was concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain the crude product. This crude product was slurried with n-heptane (150 mL), stirred at 15°C for 12 hours, filtered, and the filter cake dried to obtain compound 7.

[0113] 1 H NMR (400MHz, CDCl3) δ 8.13 (s, 1H), 8.08 (d, J = 6.5Hz, 1H), 7.53 (d, J = 9.5Hz, 1H), 7.12 (s, 1H), 4.77 (br s, 2H), 4.45 (q, J = 7.0Hz, 2H), 2.85 (br s, 1H), 1.74 (s, 6H), 1.41 (t, J=7.0Hz, 3H).

[0114] Step 6: Preparation of compound 8

[0115] Compound 7 (4.2 g, 11.56 mmol, 1 eq) was dissolved in tetrahydrofuran (250 mL) and protected with nitrogen. Wet Pd / C (5 g, 10% content) was added. The gas was replaced three times with a hydrogen balloon (15 psi). The mixture was stirred at 15°C for 4 hours. After completion of the reaction, the reaction solution was filtered through celite. The filtrate was directly dried to yield compound 8.

[0116] 1 H NMR (400MHz, DMSO-d6) δ7.87-7.77 (m, 1H), 7.51 (d, J = 11.5Hz, 1H), 7.10 (s, 1H), 6.87 (d, J = 7.0Hz, 1H ), 6.60 (s, 2H), 5.72 (s, 2H), 5.51 (s, 1H), 4.28 (q, J=7.4Hz, 2H), 1.52 (s, 6H), 1.32 (t, J=7.4Hz, 3H).

[0117] Step 7: Preparation of compound of formula (I)

[0118] Compound 8 (3.8 g, 11.40 mmol, 1 eq) was dissolved in tetrahydrofuran (80 mL) and cooled to 0°C under nitrogen. Methylmagnesium bromide (3 M, 75.99 mL, 20 eq) was added dropwise to the reaction solution, and the mixture was stirred at 0°C for 2 hours. After completion, water (100 mL) was slowly added to quench the reaction, and the mixture was diluted with saturated aqueous ammonium chloride (100 mL). After completion, the mixture was extracted with ethyl acetate (200 mL x 3). The organic phase was concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 2:1 to 1:1) to obtain the compound of formula (I).

[0119] 1 H NMR (400MHz, DMSO-d6) δ7.78 (s, 1H), 7.06 (s, 1H), 6.92 (d, J=12.5Hz, 1H), 6.68 (d , J=7.5Hz, 1H), 5.64 (s, 2H), 5.45 (s, 1H), 5.36 (s, 2H), 5.31 (s, 1H), 1.52 (s, 12H).

[0120] Example 2: Preparation of Crystal Form A of the Compound of Formula (I)

[0121] Weigh 100 mg of the compound of formula (I) into a glass vial and add 1 mL of MeOH. The solution becomes clear. After adding a magnetic stirrer and allowing to stand at room temperature, the sample is stirred at 50°C for 48 hours, then cooled to 25°C and stirred for 4 hours. The clear solution is evaporated to dryness to obtain Form A of the compound of formula (I).

[0122] Example 3: Preparation of Form B of the compound of formula (I)

[0123] The compound of formula (I) was subjected to a TGA heating experiment to 160° C., and after cooling, the sample was tested by XRPD to obtain the crystal form B of the compound of formula (I).

[0124] Weigh 100-150 mg of the compound of formula (I) into a glass vial. Add 1 mL of each solvent as shown in Table 5 below to form a suspension. Add a magnetic stirrer and allow to stand at room temperature. Stir the sample at 50°C for 48 hours, then cool to 25°C and stir for 4 hours. Filter and dry to obtain Form B of the compound of formula (I).

[0125] Table 5

[0126] serial number Compound mass solvent Volume ratio Crystal form 1 150mg ethanol / B crystal form 2 150mg Ethyl acetate / B crystal form 3 100mg Acetonitrile / B crystal form 4 100mg Methyl tert-butyl / B crystal form 5 150mg Methyl tert-butyl ether: methanol 95∶5 B crystal form 6 100mg Methyl tert-butyl ether: ethanol 95∶5 B crystal form 7 150mg Acetonitrile:water 95∶5 B crystal form

[0127] Example 4: Preparation of Form C of the compound of formula (I)

[0128] Weigh 50 mg of the compound of formula (I) into a glass vial and add 1 mL of n-heptane to form a suspension. After adding a magnetic stirrer and allowing the suspension to stand at room temperature, stir the sample at 50°C for 48 hours, then cool to 25°C and stir for 4 hours. Filter and dry the suspension to obtain Form C of the compound of formula (I).

[0129] Example 5: Preparation of Crystal Form D of the Compound of Formula (II)

[0130] Approximately 50 mg of the compound of formula (I) was weighed and added to a 4.0 mL glass vial. An appropriate amount of methanol was added and centrifuged. The clarified solution was sealed with tin foil, pierced with a small hole, and allowed to evaporate slowly at room temperature. After evaporation, the solution was transferred to a vacuum drying oven at 40°C and dried overnight to obtain the crystalline form D of the compound of formula (II).

[0131] Experimental Example 1: In vitro aldehyde capture ability experiment

[0132] 1. Experimental Purpose and Process

[0133] Purpose: Dry eye is caused by inflammation within the eye, which produces aldehydes in the body. If these aldehydes are not promptly eliminated, they accelerate the symptoms of inflammation and worsen the dry eye. This experiment simulates the in vivo environment and selects relatively optimal compounds based on their complexing ability with aldehydes in the body.

[0134] Procedure: Dissolve sulfobutyl-B-cyclodextrin (310 mg) in phosphate buffer (1.25 ml) to prepare a solution.

[0135] At room temperature, nonanal (5.0 mg, 32 μmol, 1.0 eq) and triolein (300 mg) were added to the reaction flask. After adding the above-prepared solution, linoleic acid (300 mg) was added, and finally a solution of the compound of the present invention (32 μmol, 1.0 eq) in dimethyl sulfoxide (0.15 ml) was added. The reaction solution was reacted at 20-23°C.

[0136] After stirring and reacting for 10 minutes, 100 minutes, 200 minutes, and 300 minutes, the solution was allowed to stand for 2 minutes to separate the layers and then sampled for high performance liquid chromatography (HPLC) detection.

[0137] Sampling method: Use a pipette to sample 25 μl of the upper emulsion layer and 50 μl of the lower aqueous phase, and dilute with 1 ml of methanol.

[0138] 2. Experimental Results

[0139] Nonanal has weak UV absorption at a wavelength of 254nm, and has little effect on the overall content of the complex product. Therefore, the percentage content of the complex at 254nm by HPLC is compared to observe the ability of the compound's aldehyde to capture the complexed aldehyde. Figure 4 and Table 6:

[0140]

[0141] Table 6 Statistics of compound aldehyde capture ability test results

[0142] Compound number Linear equations Slope <![CDATA[Regression coefficient R 2 > Area under the curve AUC (min. conversion rate) Compound of formula (I) y=0.1002x+7.7455 0.1002 <![CDATA[R 2 =0.9686]]> 6944

[0143] The HPLC analysis method is detailed in Table 7, which is XBRIGE 2.5 μm, 3.0*100 mm 5-80 CD_XBEH_12 min_0.8.1 cm

[0144] Table 7

[0145]

[0146]

[0147] After 300 minutes of reaction, the retention times of 6.689 minutes, 6.787 minutes, 6.966 minutes, and 7.102 minutes are the absorption peaks of the product complexed with one molecule of aldehyde; the retention times of 8.905 minutes, 9.010 minutes, and 9.075 minutes are the absorption peaks of the product complexed with two molecules of aldehyde.

[0148] The percentage of the specific complex product at 254 nm is calculated as the sum of the percentages of the absorption peaks at the above retention times, i.e., (1.839 + 1.715 + 14.993 + 13.029)% + (1.004 + 2.212 + 1.247)% = 36.039%

[0149] The specific HPLC data of the percentage content of the complex product of the compound of the present invention is as follows in Table 8:

[0150] Table 8

[0151]

[0152] Conclusion: The compound of formula (I) has very significant ability and speed to complex aldehydes.

[0153] Experimental Example 2: In vitro evaluation

[0154] Objective: To study the inhibitory effect of compounds on human liver microsomal cytochrome P450

[0155] Experimental procedure: The inhibitory activity of the compound on human liver microsomal cytochrome P450 (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) was studied. Mixed human liver microsomes were selected as the CYP450 enzyme source. Different concentrations of the compound (10, 5, 1.5, 0.5, 0.15, 0.05, 0.015 mM) were incubated with the probe substrates of the five CYP enzymes (two substrates for CYP3A4) and the cofactor (NADPH). The IC values of the compound for each CYP enzyme inhibition were determined. 50 The test results are shown in Table 9 below:

[0156] Table 9 Inhibitory effect of the compounds of the present invention on human liver microsomal cytochrome P450 isoenzymes

[0157] Compound number CYP1A2 / CYP2C9 / CYP2C19 / CYP2D6 / CYP3A4(μM) Compound of formula (I) ＞50 / ＞50 / ＞50 / ＞50 / ＞50

[0158] Conclusion: The compound of formula (I) has high safety and low possibility of drug interaction.

[0159] Experimental Example 3: Pharmacokinetic Evaluation of Compounds one,

[0161] Experimental purpose: To study the pharmacokinetics of the test compound in beagle dogs

[0162] Experimental materials: Beagle dogs (male, 8-11 kg, 6 months or older, Mas)

[0163] Experimental operation:

[0164] The pharmacokinetic profile of the compound following intravenous administration in rodents was tested using a standard protocol. The candidate compound was prepared as a clear solution and administered as a single intravenous injection at 1 mg / kg to beagle dogs. The intravenous injection vehicle was 10% hydroxypropyl β-cyclodextrin in water. Whole blood was collected at 0.033, 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-inoculation. The supernatant was separated and plasma samples were obtained by centrifugation at 3000 r / min for 10 minutes. 20 μL of plasma sample was mixed with 400 μL of acetonitrile solution containing an internal standard to precipitate protein. After centrifugation, 2 μL of the supernatant was injected. LC-MS / MS analysis was performed to quantify plasma concentrations, and pharmacokinetic parameters such as clearance, half-life, and area under the concentration-time curve (AUC) were calculated.

[0165] The experimental results are shown in Table 10:

[0166] Table 10 Pharmacokinetic test results

[0167]

[0168] Conclusion: The compound of formula (I) has a high clearance rate, a moderate half-life, and good pharmacokinetic characteristics. two,

[0170] Experimental purpose: Pharmacokinetic study of the test compound in cynomolgus monkeys

[0171] Experimental materials: Cynomolgus macaques (male, 2.5-4 kg, 2 years or older, Jingang, Hainan)

[0172] Experimental operation:

[0173] The pharmacokinetic profile of the compound following intravenous administration was tested in monkeys using a standard protocol. A single 1 mg / kg intravenous injection of the candidate compound was administered to cynomolgus monkeys as a clear solution. The intravenous injection vehicle was 10% hydroxypropyl β-cyclodextrin in water. Whole blood was collected at 0.033, 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours after incubation. The supernatant was separated and plasma samples were obtained by centrifugation at 3000 r / min for 10 minutes. 20 μL of plasma sample was mixed with 400 μL of acetonitrile solution containing an internal standard to precipitate protein. After centrifugation, 2 μL of the supernatant was injected. LC-MS / MS analysis was performed to quantify plasma concentrations, and pharmacokinetic parameters such as clearance, half-life, and area under the concentration-time curve (AUC) were calculated.

[0174] The experimental results are shown in Table 11:

[0175] Table 11 Pharmacokinetic test results

[0176]

[0177] Conclusion: The compound of formula (I) has a high clearance rate, a moderate half-life, and good pharmacokinetic characteristics.

[0178] Experimental Example 4: In vivo pharmacokinetic studies

[0179] Experimental purpose: To determine the ratio of corneal and plasma drug concentrations after eye drops administration in rats

[0180] Experimental materials: Male SD rats, 200-300 g, 7-9 weeks old, purchased from Shanghai Lingchang

[0181] Experimental procedure: The candidate compound was prepared into a clear solution and administered to SD rats by eye drops. The eye drop solvent was a 10% hydroxypropyl β-cyclodextrin aqueous solution, and the eye drop drug concentration was 5 mg / mL. Corneas and whole blood were collected at 1 and 4 hours, respectively. The corneas were homogenized using 15mM phosphate buffer (PBS): MeoH (2:1, v:v) buffer, and the whole blood samples were centrifuged at 3000r for 10 minutes to separate the supernatant plasma samples. 20μL of plasma samples and homogenate samples were mixed with 400μL of acetonitrile solution containing internal standard to precipitate proteins, centrifuged, and 2μL of the supernatant was injected. The blood drug concentration was quantitatively analyzed by LC-MS / MS analysis method. The drug concentrations in the cornea and plasma were detected at different time points, and the cornea / plasma ratio was calculated.

[0182] The experimental results are shown in Table 12:

[0183] Table 12 Pharmacokinetic test results

[0184]

[0185] Conclusion: Through the DMPK test, it can be seen that the compound of formula (I) can enter the cornea through eye drop administration and exert a pharmacological effect. The cornea / plasma ratio of the compound of formula (I) is high, with a ratio greater than 10. The pharmacological effect is on the cornea, with good targeting.

[0186] Experimental Example 5: In vivo efficacy evaluation

[0187] Purpose of the test:

[0188] Subcutaneous scopolamine injection can induce dry eye in mice. Tear testing and corneal fluorescein staining scores can reveal decreased tear secretion and inflammatory infiltration. This can also be used to predict whether the model will achieve the desired severity at the initial stage of model establishment.

[0189] Experimental Design:

[0190] · Select 20 animals from 25 female C57BL / 6J mice and randomly divide them into 4 groups according to body weight using Provantis or Excel, with 4 animals in each group. The grouping will also refer to the tear test and corneal fluorescence staining score results of each animal before the experiment.

[0191] On days 1 to 12 of the experiment, mice were subcutaneously injected with scopolamine hydrobromide 4 times a day (3±0.5h) to establish a dry eye model in mice, with a dose of 0.1 mL / mouse / time.

[0192] From Days 1 to 13 of the experiment, animals were administered eye drops four times daily (3 ± 0.5 hours) at 3 μL / eye (Day 13, no administration in Group 1). Each subcutaneous injection of scopolamine hydrobromide was administered before eye drops (except for examinations on Days 7 and 12).

[0193] Before modeling, all experimental animals were subjected to tear tests and corneal fluorescent staining scores on Day 7 and Day 12.

[0194] Scoring criteria for corneal fluorescence staining: The animal's cornea is divided into five regions: superior, inferior, nasal, temporal, and central. Each region is scored on a scale of 0 to 3. The score for each eye is the sum of the scores for the five regions. 0: no staining; 1: mild staining with fewer than 5 punctate staining; 2: moderate staining with punctate staining but no patchy staining; 3: severe staining with obvious fluorescent patches.

[0195] Tear tests were performed on all experimental animals 30 minutes after the second administration on Day 7 and Day 12, and corneal fluorescence staining scores were performed on all experimental animals 30 minutes after the third administration on Day 7 and Day 12. The experimental conditions are shown in Table 13:

[0196] Table 13 Experimental protocol for the efficacy of subcutaneous injection of scopolamine in mice

[0197]

[0198]

[0199] Test results: see Figure 5 and Figure 6 (Note: In the accompanying figures, * indicates P≤0.5, ** indicates P≤0.01, and *** indicates P≤0.001).

[0200] Conclusion: Under the experimental conditions, it can be seen that the compound of formula (I) can improve tear secretion and corneal inflammation, showing a pharmacological effect in improving dry eye.

Claims

1. Crystal form A of the compound of formula (I), characterized in that Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 16.36±0.20°, 21.23±0.20°; 2. The crystal form A according to claim 1, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 8.20±0.20°, 16.36±0.20°, 17.73±0.20°, 20.69±0.20°, 21.23±0.20°, 22.51±0.20°, and 28.36±0.20°.

3. The crystal form A according to claim 2, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 7.03±0.20°, 8.20±0.20°, 14.16±0.20°, 16.36±0.20°, 17.73±0.20°, 19.42±0.20°, 20.69±0.20°, 21.23±0.20°, 22.51±0.20°, and 28.36±0.20°.

4. The crystal form A according to claim 3, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 7.03°, 8.20°, 9.19°, 12.53°, 13.09°, 14.16°, 15.78°, 16.36°, 17.73°, 18.46°, 19.42°, 20.15°, 20.69°, 21.23°, 22.51°, 23.45°, 24.02°, 24.86°, 27.36°, 28.36°, 31.36°, 32.41°, and 38.61°.

5. The crystal form A according to claim 4, whose XRPD pattern is shown in Figure 1.

6. Crystal form B of the compound of formula (I), characterized in that Its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 14.28±0.20°, 14.78±0.20°, 20.18±0.20°; 7. The crystal form B according to claim 6, having an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 10.08±0.20°, 12.60±0.20°, 14.28±0.20°, 14.78±0.20°, 20.18±0.20°, 21.32±0.20°, 26.08±0.20°, and 27.69±0.20°.

8. The crystal form B according to claim 7, having an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ angles: 10.08±0.20°, 12.60±0.20°, 14.28±0.20°, 14.78±0.20°, 20.18±0.20°, 21.32±0.20°, 25.26±0.20°, 26.08±0.20°, 27.69±0.20°, and 29.63±0.20°.

9. The crystal form B according to claim 8, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.09°, 10.08°, 12.60°, 14.28°, 14.78°, 15.13°, 16.08°, 16.38°, 17.51°, 18.25°, 20.18°, 20.56°, 21.12°, 21.32°, 22.65°, 23. 3.85°, 24.15°, 24.77°, 25.26°, 26.08°, 26.34°, 26.64°, 27.69°, 28.94°, 29.12°, 29.63°, 30.43°, 32.16°, 32.53°, 33.34°, 33.74°, 34.74°, 35.13°, 36.20°, 37.95°, 38.24°.

10. The crystal form B according to claim 9, whose XRPD pattern is shown in Figure 2.

11. Use of the crystal form according to any one of claims 1 to 10 in the preparation of a medicament for treating diseases associated with aldehyde scavengers.

12. The use according to claim 11, characterized in that The drug for the aldehyde scavenger-related condition is a drug for dry eye.

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