Polymorphic form of benzo [c] chroman compound and preparation method and application thereof

Abstract

The invention discloses a polymorphic form of a benzo [c] chroman compound as well as a preparation method and application thereof, and particularly provides a polymorphic form of a compound as shown in a formula I as well as a preparation method and application thereof. The compound shown in the formula I serves as a cathepsin C inhibitor, and the crystal form of the compound and the pharmaceutical composition containing the crystal form can be used for treating asthma, obstructive pulmonary disease, bronchiectasis, ANCA-related vasculitis, psoriasis, alpha1-antitrypsin deficiency, lupus nephritis, diabetes, inflammatory bowel disease, rheumatic arthritis, nasosinusitis, hidradenitis suppurativa or cancer. # imgabs0 #

Description

Polymorphs of benzo[c]chroman compounds and their preparation methods and uses Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a polymorph of a benzo[c]chroman compound and a preparation method and use thereof. Background Art

[0002] Cathepsins are a class of proteolytic enzymes found widely within the lysosomes of various tissue cells. Based on their structure and catalytic activity, cathepsins are divided into three categories: serine proteases (cathepsins A and G), aspartic proteases (cathepsins D and E), and cysteine ​​proteases. Cysteine ​​proteases are the largest family of cathepsins and include 11 proteases: cathepsins B, C, F, H, K, L, O, S, W, V, and Z.

[0003] Cathepsin C, also known as dipeptidyl peptidase I or "DPP1," is constitutively expressed in many tissues, with the highest levels in the lung, kidney, liver, and spleen. Recently published studies have begun to describe the role of cathepsin C in certain inflammatory processes. For example, Adkison et al., J Clin Invest. 2002 Feb;109(3):363-71; Tinh et al., Archives of Biochemistry and Biophysics. 2002 403:160-170. These studies suggest that cathepsin C is co-expressed with certain serine proteases in granules and functions to process the precursor forms of these proteases into active forms, which are then released from inflammatory cell granules recruited to sites of inflammation. Once activated, these proteases have many functions, including the degradation of various extracellular matrix components, which together can propagate tissue damage and chronic inflammation.

[0004] WO 2004 / 110988 relates to certain nitrile derivatives and their use as DPP1 inhibitors.

[0005] WO 2009 / 074829 relates to peptidyl nitriles and their use as DPP1 inhibitors.

[0006] WO 2010 / 128324 relates to α-aminoamide nitriles and their use as DPP1 inhibitors.

[0007] WO 2012 / 119941 relates to peptidyl nitrile compounds and their use as DPP1 inhibitors.

[0008] WO 2013 / 041497 relates to N-[1-cyano-2-(phenyl)ethyl]-2-azabicyclo[2.2.1]heptane-3-carboxamides and their use as DPP1 inhibitors.

[0009] WO 2001 / 096285 and WO 2003 / 048123 relate to β-aminoamide nitriles having inhibitory activity against cysteine ​​proteases.

[0010] WO 2015 / 110826 relates to α-aminoamide nitriles and their use as DPP1 inhibitors.

[0011] WO 2022 / 117059 provides a cathepsin C inhibitor, whose chemical name is (S)-N-((S)-1-cyano-2-(8-cyano-2-fluoro-6H-benzo[c]chroman-3-yl)ethyl)-1,4-oxazepane-2-carboxamide, and has the structure shown in Formula I.

[0012] The crystal form 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 morphologies. Generally speaking, amorphous pharmaceutical products lack a regular crystal structure and often exhibit other drawbacks, such as poor product stability, fine crystallization, difficulty in filtration, susceptibility to agglomeration, and poor flowability. Polymorphic drug forms impose varying requirements on product storage, production, and scale-up. Therefore, in-depth research into the crystal forms of these compounds is essential to improve their various properties.

[0013] Summary of the Invention

[0014] The present disclosure provides a crystalline form A of a compound represented by formula I, wherein an X-ray powder diffraction pattern expressed in terms of a diffraction angle 2θ has characteristic peaks at 8.910, 10.273, 15.650, 18.617, 17.869, and 19.526, and the 2θ angle error range is ±0.20.

[0015] In some embodiments, the crystal form A of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.910, 10.273, 13.730, 15.650, 17.869, 18.617, 19.526 and 23.452, and the error range of the 2θ angle is ±0.20.

[0016] In some embodiments, the crystal form A of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.910, 10.273, 13.730, 15.650, 17.869, 18.617, 19.526, 20.785, 22.193, 23.452, 25.049 and 26.590, and the error range of the 2θ angle is ±0.20.

[0017] In some embodiments, the crystal form A of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in FIG1 .

[0018] In some embodiments, the crystal form A of the compound represented by Formula I has a DSC spectrum with endothermic peaks at 116°C and 208°C, and the error range of the endothermic peaks is ±2°C.

[0019] The present disclosure also provides a method for preparing crystal form A of the compound represented by formula I, the method comprising:

[0020] a) mixing the compound represented by formula I with solvent A;

[0021] b) stirring;

[0022] The solvent A is selected from water, C 1-4 Alcohol solvents, water and C 1-4 A mixed solvent of alcohol solvents; said C 1-4 Preferred alcohol solvents are methanol and ethanol.

[0023] In some embodiments, in the preparation method of Form A of the compound of Formula I, each milligram of the compound of Formula I is mixed with 0.01-0.05 ml of solvent A; more preferably, each milligram of the compound of Formula I is mixed with 0.01-0.03 ml of solvent A.

[0024] In some embodiments, in the preparation method of crystal form A, the temperature during stirring is room temperature or 50°C.

[0025] In some embodiments, in the preparation method of crystal form A, the mixed solvent of water and an alcohol solvent is a mixed solvent of water and methanol, wherein the molar ratio of water to the methanol is 0.10-0.95, preferably 0.14, 0.26, 0.37, 0.47, 0.57, 0.66, 0.74, 0.82, or 0.90.

[0026] The present disclosure also provides a crystal form B of the compound represented by the above formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 6.770, 11.201, 12.989, 14.931 and 20.817, and the error range of the 2θ angle is ±0.20.

[0027] In some embodiments, the crystal form B of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 6.770, 11.201, 12.989, 14.931, 20.817 and 25.710, and the error range of the 2θ angle is ±0.20.

[0028] In some embodiments, the crystal form B of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 6.770, 11.201, 12.989, 14.931, 20.817, 25.710, 31.066, 32.046 and 32.913, and the error range of the 2θ angle is ±0.20.

[0029] In some embodiments, the crystalline form B of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ having characteristic peaks at 6.770, 11.201, 12.989, 14.931, 15.851, 17.228, 20.817, 22.313, 22.773, 25.710, 28.192, 31.066, 32.046 and 32.913, and the 2θ angle error range is ±0.20.

[0030] In some embodiments, the crystal form B of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in FIG6 .

[0031] In some embodiments, the crystal form B of the compound represented by Formula I has a DSC spectrum with an endothermic peak at 210°C, and the error range of the endothermic peak is ±2°C.

[0032] The present disclosure also provides a method for preparing the crystal form B of the compound represented by Formula I, the method comprising:

[0033] a) mixing the compound of formula I with solvent B;

[0034] b) stirring;

[0035] The solvent B is selected from nitromethane and acetonitrile.

[0036] In some embodiments, in the method for preparing the crystal form B of the compound of formula I, every 100 mg of the compound of formula I is mixed with 0.5-5 ml of solvent B; more preferably, every 100 mg of the compound of formula I is mixed with 0.5-2 ml of solvent B.

[0037] In some embodiments, in the method for preparing the crystal form B of the compound of Formula I, every 100 mg of the compound of Formula I is mixed with 1 ml of solvent B.

[0038] In some embodiments, in the preparation method of crystal form B, the stirring temperature is room temperature or 50°C.

[0039] The present disclosure also provides a crystal form C of the compound represented by the above formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 7.385, 10.171, 12.687, 15.902 and 19.645, and the error range of the 2θ angle is ±0.20.

[0040] In some embodiments, the crystal form C of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 6.012, 7.385, 8.275, 10.171, 12.687, 15.120, 15.902 and 19.645, and the error range of the 2θ angle is ±0.20.

[0041] In some embodiments, the crystalline form C of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ having characteristic peaks at 6.012, 7.385, 8.275, 10.171, 12.687, 15.120, 15.902, 19.645, 25.539 and 26.382, and the 2θ angle error range is ±0.20.

[0042] In some embodiments, the crystalline form C of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ having characteristic peaks at 6.012, 7.385, 8.275, 10.171, 12.687, 13.720, 15.120, 15.902, 16.842, 19.645, 20.739, 25.539, 26.382, 27.272 and 30.887, and the 2θ angle error range is ±0.20.

[0043] In some embodiments, the Form C of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in FIG10 .

[0044] In some embodiments, the crystal form C of the compound represented by Formula I has a DSC spectrum with an endothermic peak at 209°C, and the error range of the endothermic peak is ±2°C.

[0045] The present disclosure also provides a method for preparing Form C of the compound represented by Formula I, the method comprising:

[0046] a) mixing the compound represented by formula I with solvent C;

[0047] b) stirring;

[0048] The solvent C is selected from N,N-dimethylformamide, acetone, butanone, tetrahydrofuran and 1,2-dimethoxyethane.

[0049] In some embodiments, in the method for preparing the crystalline form C of compound represented by Formula I, every 100 mg of compound represented by Formula I is mixed with 0.5-5 ml of solvent C; more preferably, every 100 mg of compound represented by Formula I is mixed with 0.5-2 ml of solvent C, and most preferably, every 100 mg of compound represented by Formula I is mixed with 1 ml of solvent C.

[0050] In some embodiments, in the preparation method of Form C, the stirring temperature is room temperature or 50°C.

[0051] The present disclosure also provides a D-type crystalline form of the compound represented by the above-mentioned formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.477, 10.451, 13.569, 15.146, 16.335 and 17.275, and the error range of the 2θ angle is ±0.20.

[0052] In some embodiments, the D form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275 and 18.775, and the error range of the 2θ angle is ±0.20.

[0053] In some embodiments, the D form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275, 18.775, 20.298, 23.990, 26.006 and 28.135, and the error range of the 2θ angle is ±0.20.

[0054] In some embodiments, the D form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275, 18.775, 20.298, 23.093, 23.990, 24.921, 26.006, 27.047, 28.135 and 33.461, and the error range of the 2θ angle is ±0.20.

[0055] In some embodiments, the D crystal form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in FIG14 .

[0056] In some embodiments, the D crystal form of the compound represented by Formula I has an endothermic peak at 211° C. in the DSC spectrum, and the error range of the endothermic peak is ±2° C.

[0057] The present disclosure also provides a method for preparing the D crystal form of the compound represented by Formula I, the method comprising:

[0058] a) mixing the compound represented by formula I with solvent D;

[0059] b) stirring;

[0060] The solvent D is selected from ethyl acetate, 1,2-xylene, toluene, 1,4-dioxane and hexane.

[0061] In some embodiments, in the preparation method of the D crystal form of the compound represented by Formula I, every 100 mg of the compound represented by Formula I is mixed with 0.5-5 ml of solvent D; more preferably, every 100 mg of the compound represented by Formula I is mixed with 0.5-2 ml of solvent D, and preferably every 100 mg of the compound represented by Formula I is mixed with 1 ml of solvent D.

[0062] In some embodiments, in the preparation method of Form D, the stirring temperature is room temperature or 50°C.

[0063] In some embodiments, the methods described herein further comprise filtering, washing, and / or drying.

[0064] The present disclosure also provides a crystalline form E of the compound represented by the above formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.685, 15.433, 16.654, 17.526 and 18.779, and the error range of the 2θ angle is ±0.20.

[0065] In some embodiments, the E form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.685, 10.681, 13.773, 14.600, 15.433, 16.654, 17.526 and 18.779, and the error range of the 2θ angle is ±0.20.

[0066] In some embodiments, the E form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.685, 10.681, 13.773, 14.600, 15.433, 16.654, 17.526, 18.779, 19.393, 20.610, 21.653, 23.319 and 24.151, and the error range of the 2θ angle is ±0.20.

[0067] In some embodiments, the E form of the compound represented by Formula I, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 7.233, 8.685, 10.681, 13.773, 14.600, 15.433, 16.654, 17.526, 18.779, 20.610, 21.653, 23.319, 24.151, 25.111 and 26.192, and the error range of the 2θ angle is ±0.20.

[0068] In some embodiments, the E crystal form of the compound represented by Formula I has an X-ray powder diffraction pattern represented by a diffraction angle 2θ as shown in FIG18 .

[0069] The present disclosure also provides a method for preparing the E crystal form of the compound represented by Formula I, which comprises the step of drying or heating the compound represented by Formula I, wherein the drying or heating temperature is preferably 100 to 200°C.

[0070] In some embodiments, the preparation method of the E crystal form of the compound represented by Formula I is to heat the solid compound represented by Formula I to above 100°C, preferably to heat the solid compound represented by Formula I to above 120°C, and more preferably to heat the solid compound represented by Formula I to above 150°C.

[0071] The present disclosure also provides a pharmaceutical composition, which contains the above-mentioned crystal form A, crystal form B, crystal form C, crystal form D or crystal form E, and a pharmaceutically acceptable excipient.

[0072] The present disclosure also provides a method for preparing the above-mentioned pharmaceutical composition, comprising the step of mixing the above-mentioned crystal form A, crystal form B, crystal form C, crystal form D or crystal form E with a pharmaceutically acceptable excipient.

[0073] The present disclosure also relates to the use of the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D or crystalline form E, or the aforementioned composition, in the preparation of a medicament for preventing and / or treating asthma, obstructive pulmonary disease, bronchiectasis, ANCA-associated vasculitis, psoriasis, α1-antitrypsin deficiency, lupus nephritis, diabetes, inflammatory bowel disease, rheumatoid arthritis, sinusitis, hidradenitis suppurativa, or cancer.

[0074] The present disclosure also relates to a method for preventing and / or treating asthma, obstructive pulmonary disease, bronchiectasis, ANCA-associated vasculitis, psoriasis, α1-antitrypsin deficiency, lupus nephritis, diabetes, inflammatory bowel disease, rheumatoid arthritis, sinusitis, hidradenitis suppurativa, or cancer, comprising administering a therapeutically effective amount of the aforementioned Crystal Form A, Crystal Form B, Crystal Form C, Crystal Form D, or Crystal Form E, or the aforementioned composition, to a subject in need thereof.

[0075] The present disclosure also relates to the aforementioned crystalline form A, crystalline form B, crystalline form C, crystalline form D or crystalline form E, or the aforementioned composition, for use in preventing and / or treating asthma, obstructive pulmonary disease, bronchiectasis, ANCA-associated vasculitis, psoriasis, α1-antitrypsin deficiency, lupus nephritis, diabetes, inflammatory bowel disease, rheumatoid arthritis, sinusitis, hidradenitis suppurativa, or cancer.

[0076] The "2θ or 2θ angle" mentioned in the present disclosure refers to the diffraction angle, θ is the Bragg angle, and the unit is ° or degree; the error range of each characteristic peak 2θ is ±0.20 (including the case where the number exceeding 1 decimal place is rounded off), specifically -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0077] The "excipients" described in this disclosure include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent or emulsifier that has been approved by the U.S. Food and Drug Administration for use by humans or livestock animals.

[0078] The starting material used in the crystal form preparation method of the present invention can be a compound in any form, including but not limited to: amorphous, any crystal form, hydrate, solvate, etc.

[0079] The "differential scanning calorimetry or DSC" described in this disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0080] Numerical values ​​in this disclosure, such as those related to substance content, are derived from measurements and calculations, and therefore inevitably have a certain degree of error. Generally speaking, ±10% is considered within a reasonable error range. While there may be some degree of error depending on the context in which they are used, this error may not exceed ±10% and may include ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 is an XRPD pattern of Form A;

[0082] FIG2 is a DSC diagram of Form A;

[0083] FIG3 is a TGA diagram of Form A;

[0084] FIG4 is a DVS diagram of Form A;

[0085] FIG5 is an XRPD pattern of Form B before and after DVS; (the upper one is the XRPD spectrum after DVS, and the lower one is the XRPD spectrum before DVS)

[0086] FIG6 is an XRPD pattern of Form B;

[0087] FIG7 is a DSC diagram of Form B;

[0088] FIG8 is a TGA diagram of Form B;

[0089] FIG9 is a DVS diagram of Form B;

[0090] FIG10 is an XRPD pattern of Form B before and after DVS; (the upper one is the XRPD spectrum after DVS, and the lower one is the XRPD spectrum before DVS)

[0091] Figure 11 is an XRPD pattern of Form C;

[0092] FIG12 is a DSC diagram of Form C;

[0093] Figure 13 is a TGA diagram of Form C;

[0094] Figure 14 is a DVS diagram of Form C;

[0095] FIG15 is an XRPD pattern of Form C before and after DVS; (the upper one is the XRPD spectrum after DVS, and the lower one is the XRPD spectrum before DVS)

[0096] Figure 16 is an XRPD pattern of Form D;

[0097] Figure 17 is a DSC diagram of Form D;

[0098] Figure 18 is a TGA diagram of Form D;

[0099] Figure 19 is a DVS diagram of Form D;

[0100] FIG20 is an XRPD pattern of Form D before and after DVS; (the upper one is the XRPD spectrum before DVS, and the lower one is the XRPD spectrum after DVS)

[0101] Figure 21 is an XRPD pattern of Form E;

[0102] Figure 22 is a DSC diagram of Form E;

[0103] FIG23 is a TGA chart of Form E. DETAILED DESCRIPTION

[0104] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure, and are not intended to limit the essence and scope of the present disclosure.

[0105] The abbreviations used in this disclosure are explained as follows: XRPD X-ray powder diffraction DSC Differential Scanning Calorimetry TGA Thermogravimetric Analysis DVS Dynamic Water Sorption 1 H-NMR liquid nuclear magnetic hydrogen spectrum DMF N,N-dimethylformamide MEK butanone MTBE methyl tert-butyl ether THF tetrahydrofuran IPA isopropyl alcohol ACN acetonitrile MeOH methanol EOH ethanol ACT acetone EA ethyl acetate PA phosphoric acid TA tartaric acid HBr hydrobromic acid HCl hydrochloric acid

[0106] Test conditions of the instruments used in the experiments in this disclosure:

[0107] 1. X-ray Powder Diffraction (XRPD)

[0108] Instrument model: Malver Panalytical Aeris X-ray powder diffractometer

[0109] Radiation: Monochromatic Cu-Kα radiation (λ=1.54188)

[0110] Scanning mode: θ / 2θ, scanning range (2θ range): 3.5~50°

[0111] Voltage: 40kV, Current: 15mA

[0112] 2. Differential Scanning Calorimeter (DSC)

[0113] Instrument model: TA DSC250

[0114] Purge gas: nitrogen; nitrogen purge rate: 50mL / min

[0115] Heating rate: 10℃ / min

[0116] Temperature range: 25℃-300℃

[0117] 3. Thermogravimetric Analysis (TGA)

[0118] Instrument model: TA TGA550

[0119] Purge gas: nitrogen; nitrogen purge speed: 20ml / min

[0120] Heating rate: 10℃ / min

[0121] Temperature range: 30℃-350℃

[0122] 4. Dynamic moisture sorption (DVS)

[0123] The test was performed using SMS Intrinsic PLUS at 25°C, with humidity ranging from 50% to 0% to 90% in 10% increments. The judgment standard was that the mass change dM / dT for each gradient was less than 0.002%, TMAX was 360 min, and the cycle was repeated twice.

[0124] 5. The high performance liquid chromatography (HPLC) chart described in the crystal stability test of the present disclosure was collected on an Agilent 1260 Infinity II.

[0125] The HPLC conditions in the content detection method of the high performance liquid chromatography described in the present disclosure are as follows: chromatographic column: Agilent Eclipse Plus C18 4.6mm*150mm, 3.5μm; mobile phase: A: 10mmol / L sodium dihydrogen phosphate solution (pH8.0), B-ACN; flow rate: 1.0ml / min; wavelength: 210nm.

[0126] The HPLC conditions in the high performance liquid chromatography method for detecting related substances described in Example 8 of the present disclosure are as follows: chromatographic column: Agilent Eclipse Plus C18 4.6mm*150mm, 3.5μm; mobile phase: A: 10mmol / L ammonium acetate solution, B-ACN; flow rate: 1.0ml / min; wavelength: 210nm.

[0127] The conditions for the high performance liquid chromatography detection method for the isomers of Formula I in the embodiments of the present disclosure are as follows: chromatographic column CHIRALPAK IG-3 4.6 mm*250 mm 3 μm or CHIRALPAK IG-3 4.0 mm*10 mm 3 μm, mobile phase: A: n-hexane, B: anhydrous ethanol; n-hexane: anhydrous ethanol = 25:75, flow rate: 1.0 ml / min; wavelength: 254 nm.

[0128] Example 1: Preparation of the compound represented by formula I: (S)-N-((S)-1-cyano-2-(8-cyano-2-fluoro-6H-benzo[c]chroman-3-yl)ethyl)-1,4-oxazepane-2-carboxamide

[0129] Synthesis of compound I-2

[0130] Methyl 2-fluoro-5-hydroxybenzoate (4.30 g, 25.29 mmol), 4-bromo-3-(bromomethyl)benzonitrile (Compound I-1) (7.72 g, 25.30 mmol), and potassium carbonate (6.99 g, 50.58 mmol) were dissolved in N,N-dimethylformamide (50 mL) at room temperature. The reaction mixture was heated to 40°C and stirred for 12 hours. Water (300 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 2). The organic phases were combined, washed with saturated brine (100 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 9 / 1) to obtain Compound I-2. 1H NMR (400MHz, CDCl3) δ7.87(d,1H),7.73(d,1H),7.55-7.48(m,2H),7.18-7.09(m,2H),5.11(s,2H),3.95(s,3H).

[0131] Synthesis of compound I-3

[0132] Under nitrogen at room temperature, compound I-2 (2.90 g, 6.37 mmol), palladium acetate (0.14 g, 0.64 mmol), potassium carbonate (1.76 g, 12.73 mmol), and tricyclohexylphosphine tetrafluoroborate (0.23 g, 0.64 mmol) were dissolved in N,N-dimethylformamide (30 mL). The reaction mixture was heated to 120°C and stirred for 1.5 hours. After completion of the reaction, the mixture was cooled to room temperature, water (200 mL) was added, and the mixture was extracted with ethyl acetate (200 mL x 2). The organic phases were combined, washed with saturated brine (200 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 4 / 1) to afford compound I-3. 1 H NMR (400MHz, CDCl3) δ7.80-7.72(m,2H),7.60(d,1H),7.52-7.49(m,2H),5.18(s,2H),3.98(s,3H).

[0133] Synthesis of compound I-4

[0134] Compound I-3 (1.20 g, 3.81 mmol) and lithium borohydride (0.25 g, 11.47 mmol) were dissolved in tetrahydrofuran (25 mL) at room temperature. The reaction mixture was heated to 55°C and stirred for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, water (200 mL) was added, and extraction was performed with ethyl acetate (200 mL x 2). The organic phases were combined, washed with saturated brine (200 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 9 / 1) to obtain compound I-4. 1 H NMR (400MHz, CDCl3) δ7.67(s,2H),7.47(s,1H),7.38(d,1H),7.12(d,1H),5.11(s,2H),4.77(s,2H).

[0135] Synthesis of compound I-5

[0136] At room temperature, compound I-3 (450 mg, 1.59 mmol) was dissolved in dichloromethane (15 mL), followed by the dropwise addition of phosphorus tribromide (520 mg, 1.92 mmol). The reaction mixture was stirred at room temperature for 20 minutes. After completion of the reaction, water (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL x 2). The organic phases were combined, washed with saturated brine (100 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 9 / 1) to afford compound I-5. 1 H NMR (300MHz, CDCl3) δ7.73(s,2H),7.52(s,1H),7.45(d,1H),7.10(d,1H),5.17(s,2H),4.53(s,2H).

[0137] Synthesis of compound I-6

[0138] At room temperature, diphenylmethyleneaminoacetonitrile (250 mg, 1.14 mmol), compound I-5 (470 mg, 1.26 mmol), benzyltrimethylammonium chloride (22 mg, 0.12 mmol), and sodium hydroxide (91 mg, 2.3 mmol) were dissolved in a mixed solvent of dichloromethane (6 mL) and water (6 mL). The reaction mixture was heated to 35°C and stirred for 24 hours. Water (30 mL) was added, and the mixture was extracted with dichloromethane (30 mL x 2). The organic phases were combined, washed with saturated brine (20 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 4 / 1) to obtain compound I-6. MS-ESI: m / z 458.4 [M+1] + .

[0139] Synthesis of compound I-7

[0140] At room temperature, compound I-6 (520 mg, 0.90 mmol) was dissolved in tetrahydrofuran (10 mL), followed by the dropwise addition of 1 M aqueous hydrochloric acid solution (4 mL). The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, saturated aqueous sodium bicarbonate solution (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (SiO2, petroleum ether / ethyl acetate = 2 / 1) to obtain compound I-7. MS-ESI: m / z 293.9 [M+1] + .

[0141] Synthesis of compound I-8

[0142] At room temperature, compound I-7 (220 mg, 0.68 mmol), compound a (170 mg, 0.69 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (360 mg, 0.95 mmol), and N,N-diisopropylethylamine (250 mg, 1.93 mmol) were dissolved in N,N-dimethylformamide (5 mL). The reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction, compound I-8 was isolated by preparative liquid chromatography (C18, acetonitrile / water system). MS-ESI: m / z 465.1 [M-56+1] + .

[0143] Synthesis of compound I-9

[0144] Compound I-8 (280 mg, 0.51 mmol) was subjected to chiral separation (chromatographic column: chiralpak IE, 250*25 mm, 5 μm; mobile phase: n-hexane, ethanol; gradient ratio: 30% n-hexane phase; flow rate: 15 mL / min; column temperature: 30°C) to obtain compound I-9 (two diastereoisomer peaks in total, compound I-9 is the first eluting peak). 1 H NMR(300MHz,DMSO-d6)δ8.89(d,1H),8.08(d,1H),7.94-7.89(m,2H),7.83(s ,1H),7.07(d,1H),5.24-5.14(m,2H),5.09-5.07(m,1H),4.15-4.12(m,1H), 3.99-3.87(m,2H),3.63-3.56(m,2H),3.29-3.02(m,4H),1.89-1.79(m,2H),1.41-1.36(m,9H).

[0145] Synthesis of Compound I

[0146] Compound I-9 (85 mg, 0.16 mmol) was dissolved in formic acid (1 mL) at room temperature, and the reaction mixture was heated to 40°C and stirred for 1 hour. After completion of the reaction, preparative liquid chromatography (C18, ammonium bicarbonate / acetonitrile / water system) afforded Compound I. XRPD analysis revealed the compound to be amorphous.

[0147] MS-ESI: m / z 421.2[M+1] + . 1H NMR(400MHz,DMSO-d6)δ8.69(d,1H),8.05(d,1H),7.89-7.86(m,2H),7.80(s,1H),7.03(d,1H),5.20-5.12(m,2H),5.09-5.02(m,1H),4.00-3.9 7(m,1H),3.88-3.83(m,1H),3.77-3.68(m,1H),3.25-3.14(m,2H),3.05 -2.98(m,1H),2.82-2.72(m,1H),2.62-2.53(m,2H),1.80-1.66(m,2H).

[0148] Example 2: Biological evaluation of the compound represented by Formula I - in vitro CatC cell activity detection experiment

[0149] 1. Experimental Materials

[0150] 2. Experimental Procedure

[0151] Prepare complete culture medium (RPMI1640 + 10% FBS) and mix thoroughly. Resuscitate the U937 cell line, passage it for approximately two generations, and select a cell line with good growth. Aspirate the cell suspension and transfer it to a centrifuge tube. Centrifuge at 800-1000 rpm for 3-5 minutes. Discard the supernatant. Add an appropriate volume of culture medium to the centrifuge tube and gently pipette to resuspend the cells evenly. Count the cells using a Vi-Cell XR cell counter. Adjust the cell suspension to the appropriate concentration. Add the cell suspension to a 384-well plate at 25,000 μl / well. Prepare a 10 mM solution of the compound in DMSO. Dilute the compound to a 1 mM solution in DMSO and perform 10 half-log dilutions in DMSO using an HPD300. Prepare a 35 mM solution of Gly-Phe-AFC in DMSO, aliquot, and dilute Gly-Phe-AFC to 1.75 mM using serum-free culture medium. After adding the drug, incubate in the incubator for 1 hour before adding the substrate, AFC, and adding 12.5 μl to the plate. After incubation for 30 minutes, measure the plate. Place the plate in an EnSpire reader and record fluorescence at Ex = 400 nm and Em = 505 nm. Calculate the inhibition rate using the following formula: Inhibition rate (%) = (1 - (RFU compound - RFU blank) / (RFU DMSO - RFU blank)) × 100%. Use XLFit to plot the inhibition rate curve and calculate the IC. 50 The 4-parameter model [fit = (A + ((BA) / (1 + ((C / x)^D))))] was used.

[0152] The inhibition of the activity of the compound of formula I disclosed in the present invention on CatC cells in vitro was determined by the above test, and the measured IC50 The value is 6.1nM, which has a significant inhibitory effect on CatC cell activity.

[0153] Example 3: Preparation of Form A of the Compound of Formula I

[0154] 10 mg of the compound represented by Formula I was added to 0.2 mL of solvent, and Formula I was kept in the solvent to form a slurry. The mixture was stirred at room temperature (about 25° C.) or 50° C. for one week, and filtered. The obtained solid was detected by XRPD to be free crystalline Form A, as shown in Table 1 below.

[0155] Table 1 Solvent selection and XRPD test results for preparation of Form A by beating crystallization

[0156] X-ray powder diffraction analysis identified the product as Form A. The XRPD spectrum is shown in Figure 1, and the positions of its characteristic peaks are shown in Table 2. The DSC spectrum, shown in Figure 2, exhibits endothermic peaks at 116°C and 208°C. The TGA spectrum, shown in Figure 3, shows a 2% weight loss before 100°C and rapid weight loss above 200°C.

[0157] Table 2 XRPD characteristic diffraction peak data of Form A

[0158] DVS analysis, as shown in Figure 4, shows a moisture absorption weight gain of approximately 5.39% under accelerated experimental conditions (i.e., 80% RH). Desorption and adsorption processes were consistent across humidity ranges from 0% to 90% RH. The crystal form was re-tested after DVS analysis, and the XRPD spectrum, shown in Figure 5, shows no change in crystal form before and after DVS analysis.

[0159] Example 4: Preparation of Form B of the Compound of Formula I

[0160] About 100 mg of Compound I was added to 1 ml of solvent, and Compound I was kept slurried in the solvent. The mixture was stirred at room temperature (about 25° C.) or 50° C. for 7 days and filtered. The resulting solid was detected by XRPD to be free crystalline Form B, as shown in Table 3 below.

[0161] Table 3 Solvent selection and XRPD test results for preparation of Form B by beating crystallization

[0162] X-ray powder diffraction analysis identified the product as Form B. The XRPD spectrum is shown in Figure 6 , and the positions of its characteristic peaks are shown in Table 4 . The DSC spectrum is shown in Figure 7 , showing endothermic peaks at 114.55°C and 209.87°C. The TGA spectrum is shown in Figure 8 .

[0163] DVS analysis, as shown in Figure 9, reveals that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.93%. Under accelerated storage conditions (i.e., 80% RH), the moisture gain was approximately 1.60%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 3.87%. Furthermore, the crystal form was re-tested after DVS analysis, and the XRPD spectrum, shown in Figure 10, reveals no change in crystal form before and after DVS analysis.

[0164] Table 4 XRPD characteristic diffraction peak data of Form B

[0165] Example 5: Preparation of Form C of the Compound of Formula I

[0166] About 100 mg of Compound I was added to 1 ml of solvent, and Compound I was kept slurried in the solvent. The mixture was stirred at room temperature (about 25° C.) or 50° C. for 7 days and filtered. The obtained solid was detected by XRPD to be Form C, as shown in Table 5 below.

[0167] Table 5 Solvent selection and XRPD test results for preparation of Form C by beating crystallization

[0168] X-ray powder diffraction analysis identified the product as Form C. The XRPD spectrum is shown in Figure 11 , and the positions of its characteristic peaks are shown in Table 6 . The DSC spectrum is shown in Figure 12 , showing endothermic peaks at 75.84°C and 208.87°C. The TGA spectrum is shown in Figure 13 .

[0169] DVS analysis, as shown in Figure 14, reveals that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.65%. Under accelerated storage conditions (i.e., 80% RH), the moisture gain was approximately 0.89%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 1.16%. Furthermore, the crystal form was re-tested after DVS analysis, and the XRPD spectrum, shown in Figure 15, reveals no change in crystal form before and after DVS analysis.

[0170] Table 6 XRPD characteristic diffraction peak data of Form C

[0171] Example 6: Preparation of Form D of the Compound of Formula I

[0172] About 100 mg of Compound I was added to 1 ml of solvent, and Formula I was kept slurried in the solvent. The mixture was stirred at room temperature (about 25° C.) or 50° C. for 7 days and filtered. The resulting solid was detected by XRPD to be free crystalline Form D, as shown in Table 7 below.

[0173] Table 7 Solvent selection and XRPD test results for preparation of Form D by beating crystallization

[0174] X-ray powder diffraction analysis identified the product as Form D. The XRPD spectrum is shown in Figure 16 , and the positions of its characteristic peaks are shown in Table 8. The DSC spectrum is shown in Figure 17 , which shows an endothermic peak at 211.29°C. The TGA spectrum is shown in Figure 18 .

[0175] DVS analysis, as shown in Figure 19, reveals that under normal storage conditions (i.e., 25°C, 60% RH), the sample experienced a moisture gain of approximately 0.64%. Under accelerated storage conditions (i.e., 80% RH), the moisture gain was approximately 0.76%. Under extreme conditions (i.e., 90% RH), the moisture gain was approximately 0.84%. Furthermore, the crystal form was re-tested after DVS analysis, and the XRPD spectrum, shown in Figure 20, reveals no change in crystal form before and after DVS analysis.

[0176] Table 8 XRPD characteristic diffraction peak data of Form D

[0177] Example 7: Preparation of Form E of the Compound of Formula I

[0178] The compound of formula I, Form B, Form C, and Form D obtained in Example 1 were vacuum dried at 120° C. for 30 min and analyzed by X-ray powder diffraction. The obtained solid was found to be free Form E by XRPD.

[0179] The compound of Formula I and Forms B, C, and D of the compound of Formula I obtained in Example 1 were heated to 150°C and analyzed by X-ray powder diffraction. The resulting solid was identified as free Form E by XRPD analysis. The XRPD spectrum is shown in Figure 21 , and the positions of its characteristic peaks are shown in Table 9. The DSC spectrum is shown in Figure 22 , showing an endothermic peak at 207.44°C. The TGA spectrum is shown in Figure 23 .

[0180] Table 9 XRPD characteristic diffraction peak data of Form E

[0181] Example 8: Stability Test of Free Base Form E of the Compound Represented by Formula I

[0182] Determine whether temperature and humidity have an effect on the stability of the free base crystalline form E of the compound represented by Formula I:

[0183] The free base Form E was placed in a light-proof stabilization chamber and subjected to a series of temperature and humidity combinations for influencing factor experiments. Samples were removed after aging under various light and temperature and humidity conditions for different days and examined for related substances (impurities) by high-performance liquid chromatography. Comparison of the resulting solids with the initial free base Form E revealed no significant increase or only a slight increase in impurities. When samples were removed after aging under light conditions for different days, an increase in impurities was observed. The results are shown in Table 10.

[0184] XRPD analysis of the samples taken out on the last day of exposure to light and various temperature and humidity conditions showed that the free base crystalline form E had no change and had good stability.

[0185] Table 10 Results of factors affecting the drying treatment of free base crystal form E

Claims

1. A crystalline form A of a compound of formula I, wherein the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 8.910, 10.273, 15.650, 18.617, 17.869 and 19.526, preferably at 8.910, 10.273, 13.730, 15.650, 17.869, 18.617, 19.526 and 23.452, more preferably at 8.910, 10.273, 13.730, 15.650, 17.869, 18.617, 19.526, 20.785, 22.193, 23.452, 25.049 and 26.590, and the 2θ angle error range is ±0.

20.

2. The crystal form A according to claim 1, wherein the DSC spectrum has endothermic peaks at 116°C and 208°C, and the error range of the endothermic peaks is ±2°C.

3. A method for preparing the crystal form A according to claim 1 or 2, comprising: a) mixing the compound represented by formula I with solvent A; b) stirring; The solvent A is selected from water, C 1-4 Alcohol solvents, water and C 1-4 A mixed solvent of alcohol solvents; said C 1-4 Preferred alcohol solvents are methanol and ethanol.

4. A crystalline form B of the compound of formula I, wherein the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 6.770, 11.201, 12.989, 14.931, and 20.817, preferably at 6.770, 11.201, 12.989, 14.931, 20.817, and 25.710, more preferably at 6.770, 11.201, 12.989, 14.931, 20. 817, 25.710, 31.066, 32.046 and 32.913, most preferably 6.770, 11.201, 12.989, 14.931, 15.851, 17.228, 20.817, 22.313, 22.773, 25.710, 28.192, 31.066, 32.046 and 32.913, the 2θ angle error range is ± 0.20, 5. The crystal form B according to claim 4, wherein the DSC spectrum has endothermic peaks at 114°C and 210°C, and the error range of the endothermic peaks is ±2°C.

6. A method for preparing the crystal form B according to claim 4 or 5, comprising: a) mixing the compound represented by formula I with solvent B; b) stirring; The solvent B is selected from nitromethane and acetonitrile.

7. A crystalline form C of the compound of formula I, wherein the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 7.385, 10.171, 12.687, 15.902, and 19.645, preferably at 6.012, 7.385, 8.275, 10.171, 12.687, 15.120, 15.902, and 19.645, more preferably at 6.012, 7.385, 8.275, 10.171, 12. Characteristic peaks are present at 6.687, 15.120, 15.902, 19.645, 25.539 and 26.382, and most preferably at 6.012, 7.385, 8.275, 10.171, 12.687, 13.720, 15.120, 15.902, 16.842, 19.645, 20.739, 25.539, 26.382, 27.272 and 30.887, and the 2θ angle error range is ± 0.20, 8. The crystal form C according to claim 7, wherein the DSC spectrum has endothermic peaks at 76°C and 209°C, and the error range of the endothermic peaks is ±2°C.

9. A method for preparing the crystal form C according to claim 7 or 8, comprising: a) mixing the compound represented by formula I with solvent C; b) stirring; The solvent C is selected from N,N-dimethylformamide, acetone, butanone, tetrahydrofuran and 1,2-dimethoxyethane.

10. A crystal form D of the compound of formula I, wherein the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 8.477, 10.451, 13.569, 15.146, 16.335 and 17.275, preferably at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275 and 18.775, more preferably at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275 and 18.

775. 5, 17.275, 18.775, 20.298, 23.990, 26.006 and 28.135, preferably at 8.477, 10.451, 13.569, 15.146, 16.335, 17.275, 18.775, 20.298, 23.093, 23.990, 24.921, 26.006, 27.047, 28.135 and 33.461, the 2θ angle error range is ± 0.20, 11. The crystal form D according to claim 10, wherein the DSC spectrum has an endothermic peak at 211°C, and the error range of the endothermic peak is ±2°C.

12. A method for preparing the crystal form D according to claim 10 or 11, comprising: a) mixing the compound represented by formula I with solvent D; b) stirring; The solvent D is selected from ethyl acetate, 1,2-xylene, toluene, 1,4-dioxane and hexane.

13. A crystalline form E of a compound of formula I, wherein the X-ray powder diffraction pattern expressed in terms of diffraction angle 2θ has characteristic peaks at 8.685, 15.433, 16.654, 17.526 and 18.779, preferably at 8.685, 10.681, 13.773, 14.600, 15.433, 16.654, 17.526 and 18.779, more preferably at 8.685, 10.681, 13.773, 14.600, 15.433, 16.65 4, 17.526, 18.779, 19.393, 20.610, 21.653, 23.319 and 24.151, and most preferably have characteristic peaks at 7.233, 8.685, 10.681, 13.773, 14.600, 15.433, 16.654, 17.526, 18.779, 20.610, 21.653, 23.319, 24.151, 25.111 and 26.192, and the 2θ angle error range is ± 0.20, 14. The crystal form E according to claim 13, wherein the DSC spectrum has an endothermic peak at 207°C, and the error range of the endothermic peak is ±2°C.

15. A method for preparing the crystal form E according to claim 13 or 14, comprising the step of drying or heating the compound represented by formula I, wherein the drying or heating temperature is preferably 100°C to 200°C.

16. A pharmaceutical composition comprising the crystalline form according to any one of claims 1-2, 4-5, 7-8, 10-11 and 13-14 and a pharmaceutically acceptable excipient.

17. Use of the crystalline form according to any one of claims 1-2, 4-5, 7-8, 10-11 and 13-14 or the pharmaceutical composition according to claim 16 in the preparation of a medicament for preventing and / or treating asthma, obstructive pulmonary disease, bronchiectasis, ANCA-associated vasculitis, psoriasis, α1-antitrypsin deficiency, lupus nephritis, diabetes, inflammatory bowel disease, rheumatoid arthritis, sinusitis, hidradenitis suppurativa or cancer.

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