Crystals of polypeptide compounds containing cyano substituents and their method of manufacture

Abstract

The present invention discloses crystals of polypeptide compounds containing cyano substituents and a method for preparing the same, specifically, crystals of the compound represented by formula (I) and a method for preparing the same. [Formula 1] JPEG2025533996000028.jpg42170

Description

[Technical Field]

[0001] This application claims priority from Chinese Patent Application No. 2022112639720, filed on October 14, 2022. This application cites the above Chinese patent application in its entirety.

[0002] The present invention relates to a crystal of a polypeptide compound containing cyano substituents and a method for producing the same, and more particularly to a crystal of a compound represented by formula (I) and a method for producing the same. [Background technology]

[0003] The novel coronavirus disease 2019 (COVID-19) emerged in December 2019 and rapidly developed into a pandemic, severely impacting the global public health system and the global economy. The pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), shares a very high RNA genome similarity (approximately 80%) with SARS-CoV-1, which led to the SARS outbreak in 2003. In response to the spread and severity of SARS-CoV-2 infection, many countries and organizations have made significant efforts to develop preventative and therapeutic strategies, including vaccine development and large-scale vaccination, as well as the development and use of several therapeutic drugs.

[0004] Coronaviruses are enveloped, single-stranded, positive-stranded RNA viruses that encode structural and nonstructural proteins to facilitate viral entry into and replication within the host. The nonstructural protein 3CL (3-chymotrypsin-like protease) protease plays a key role in the viral replication cycle, and its main function is to hydrolyze two polyproteins expressed by the virus. Sequence analysis has shown that 3CL protease may be an important target for drug design.

[0005] The currently available 3CL protease inhibitor is Pfizer's Nilmatrervir tablets, which can reduce hospitalization and mortality rates by 89% and 70%, respectively, in high-risk and low-risk groups. It has been approved for sale or emergency use in the United States and many other countries. Currently, numerous 3CL proteases are undergoing various stages of clinical trials both domestically and internationally. Therefore, the development of 3CL protease inhibitors is particularly important for the treatment of 2019 novel coronavirus and other coronavirus infections. Summary of the Invention

[0006] The present invention provides a type A crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 10.655±0.200°, 11.988±0.200°, 16.055±0.200°, 18.356±0.200°, and 20.083±0.200°. [ka]

[0007] In some embodiments of the present invention, the A-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum expressed in 2θ angles, which contains at least 6, 7, or 8 diffraction peaks selected from 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, and 20.083±0.200°.

[0008] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the Type A crystal of the compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, and 20.083±0.200°.

[0009] In some embodiments of the present invention, the A-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum represented by 2θ angles of 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 13.837±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.756±0.200°, 19.756±0.200°, 20.756±0.200°, 21.756±0.200°, 22.756±0.200°, 23.756±0.200°, 24.756±0.200°, 25.756±0.200°, 26.756±0.200°, 27.756±0.200°, 28.756±0.200°, 29.756±0.200°, 30.756±0.200°, 31.756±0.200°, 32.756±0.200°, 33.756±0.200°, 34.756±0.200°, 35.756±0.200°, 36.756±0.200°, 37.756±0.200°, 38.756±0.200°, 39.756±0.200°, 40.756±0.200°, 41.756±0.200°, 42.756±0. The diffraction peaks comprise at least 12, 13, 14, 15, or 16 diffraction peaks selected from 20.356±0.200°, 20.083±0.200°, 20.839±0.200°, 21.388±0.200°, 22.379±0.200°, 24.577±0.200°, 25.104±0.200°, 26.402±0.200°, and 31.540±0.200°.

[0010] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the A-type crystal of the compound represented by formula (I) has the following 2θ angles: 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 13.837±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°. It has characteristic diffraction peaks at 0.200°, 18.356±0.200°, 20.083±0.200°, 20.839±0.200°, 21.388±0.200°, 22.379±0.200°, 24.577±0.200°, 25.104±0.200°, 26.402±0.200°, and 31.540±0.200°.

[0011] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the A-type crystal of the compound represented by formula (I) is as shown in FIG.

[0012] In some embodiments of the present invention, the peak positions, interplanar distances and relative intensities of diffraction peaks in the powder X-ray diffraction (XRPD) spectrum of the A-type crystal of the compound represented by formula (I) are as shown in Table 1. [Table 1]

[0013] In some embodiments of the present invention, the A-type crystals of the compound represented by formula (I) have an endothermic peak value at 173.5°C ± 3°C in their differential scanning calorimetry curve (DSC).

[0014] In some embodiments of the present invention, the type A crystal of the compound represented by formula (I) has a differential scanning calorimetry curve as shown in FIG.

[0015] In some embodiments of the present invention, the A-type crystal of the compound represented by formula (I) exhibits a weight loss of 0.00% when measured at 150.0°C ± 3°C in a thermogravimetric analysis curve (TGA).

[0016] In some embodiments of the present invention, the thermogravimetric analysis curve of the A-type crystal of the compound represented by formula (I) is as shown in FIG.

[0017] The present invention further provides a method for producing type A crystals of the compound represented by formula (I), which comprises a step of stirring the compound represented by formula (I) in water and further comprises a step of separating the compound.

[0018] In some embodiments of the present invention, the A-type crystal of the compound of formula (I) of the present invention is (1) A method of adding water to a compound represented by formula (I) and stirring the mixture; (2) Filtration and vacuum concentration to remove water Manufactured by.

[0019] In some embodiments of the present invention, the stirring in step (1) is carried out under heated conditions, and preferably, the stirring temperature is 50°C.

[0020] The present invention further provides a B-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.202±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, and 19.917±0.200°. [ka]

[0021] In some embodiments of the present invention, the B-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum expressed in 2θ angles, which contains at least 6, 7, or 8 diffraction peaks selected from 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, 18.306±0.200°, and 19.917±0.200°.

[0022] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the compound B-type crystal represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, 18.306±0.200°, and 19.917±0.200°.

[0023] In some embodiments of the present invention, the B-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum represented by 2θ angles of 5.459±0.200°, 7.096±0.200°, 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, and 16. The diffraction peaks comprise at least 12, 13, 14, 15, or 16 diffraction peaks selected from: 483±0.200°, 18.306±0.200°, 19.488±0.200°, 19.917±0.200°, 20.681±0.200°, 21.526±0.200°, 22.409±0.200°, 24.622±0.200°, and 25.661±0.200°.

[0024] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the B-type crystal of the compound represented by formula (I) has the following 2θ angles: 5.459±0.200°, 7.096±0.200°, 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, and 15.516±0.200°. It has characteristic diffraction peaks at 200°, 16.483±0.200°, 18.306±0.200°, 19.488±0.200°, 19.917±0.200°, 20.681±0.200°, 21.526±0.200°, 22.409±0.200°, 24.622±0.200°, and 25.661±0.200°.

[0025] In some embodiments of the present invention, the B-type crystals of the compound represented by formula (I) have an X-ray powder diffraction spectrum (XRPD) as shown in FIG.

[0026] In some embodiments of the present invention, the peak positions, interplanar distances and relative intensities of diffraction peaks in the powder X-ray diffraction spectrum of type B crystals of the compound represented by formula (I) are as shown in Table 2. [Table 2]

[0027] In some embodiments of the present invention, the B-type crystals of the compound represented by formula (I) have an endothermic peak value at 128.4°C ± 3°C in their differential scanning calorimetry curve (DSC).

[0028] In some embodiments of the present invention, the B-type crystals of the compound represented by formula (I) have a differential scanning calorimetry curve as shown in FIG.

[0029] In some embodiments of the present invention, the B-type crystals of the compound represented by formula (I) show a weight loss of 9.77% when the thermogravimetric analysis curve (TGA) is at 150.0°C±3°C.

[0030] In some embodiments of the present invention, the thermogravimetric analysis curve of the B-type crystals of the compound represented by formula (I) is as shown in FIG.

[0031] In some embodiments of the present invention, the type B crystals of the compound represented by formula (I) may exist in the form of a solvate crystal.

[0032] The present invention further provides a method for producing type B crystals of the compound represented by formula (I), which comprises a step of stirring the compound represented by formula (I) in an alcohol / n-heptane mixed solvent at room temperature, and further comprises a separation step.

[0033] In some embodiments of the present invention, the B-type crystal of the compound represented by formula (I) of the present invention is (1) A method in which an alcohol / n-heptane mixed solvent is added to a compound represented by formula (I) and the mixture is stirred at room temperature; (2) Evaporation at low temperature and concentration under reduced pressure Manufactured by.

[0034] In some embodiments of the present invention, the alcohol / n-heptane mixed solvent in the above step (1) is preferably ethanol / n-heptane.

[0035] In some embodiments of the present invention, the solvent ratio of ethanol / n-heptane is preferably 1 / 4 (volume ratio).

[0036] In some embodiments of the present invention, the low temperature in step (2) above is preferably 5°C.

[0037] The present invention further provides a C-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 6.334±0.200°, 8.399±0.200°, 9.435±0.200°, 12.060±0.200°, and 18.126±0.200°. [ka]

[0038] In some embodiments of the present invention, the C-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum expressed in 2θ angles, which contains at least 6, 7, or 8 diffraction peaks selected from 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, and 18.126±0.200°.

[0039] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the C-type crystal of the compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, and 18.126±0.200°.

[0040] In some embodiments of the present invention, the C-type crystal of the compound represented by formula (I) has a powder X-ray diffraction spectrum represented by 2θ angles of 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, 12.9 The diffraction peaks comprise at least 12, 13, 14, 15, or 16 diffraction peaks selected from: 88±0.200°, 14.821±0.200°, 16.182±0.200°, 18.126±0.200°, 18.545±0.200°, 19.474±0.200°, 20.199±0.200°, 24.294±0.200°, and 25.697±0.200°.

[0041] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the C-type crystal of the compound represented by formula (I) has the following 2θ angles: 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.2 It has characteristic diffraction peaks at the following angles: 12.988±0.200°, 14.821±0.200°, 16.182±0.200°, 18.126±0.200°, 18.545±0.200°, 19.474±0.200°, 20.199±0.200°, 24.294±0.200°, and 25.697±0.200°.

[0042] In some embodiments of the present invention, the C-type crystal of the compound represented by formula (I) is characterized by having a powder X-ray diffraction spectrum as shown in FIG.

[0043] In some embodiments of the present invention, the peak positions, interplanar distances and relative intensities of diffraction peaks in the powder X-ray diffraction spectrum of C-type crystals of the compound represented by formula (I) are as shown in Table 3. [Table 3]

[0044] In some embodiments of the present invention, the differential scanning calorimetry curve of the C-type crystals of the compound represented by formula (I) has an endothermic peak value at 113.5°C ± 3°C.

[0045] In some embodiments of the present invention, the DSC spectrum of the C-type crystals of the compound represented by formula (I) is as shown in FIG.

[0046] In some embodiments of the present invention, the thermogravimetric analysis curve of the C-type crystal of the compound represented by formula (I) shows a weight loss of 3.52% at 110.0°C±3°C and a weight loss of 10.58% at 150.0°C±3°C.

[0047] In some embodiments of the present invention, the TGA spectrum of the C-type crystals of the compound represented by formula (I) is as shown in FIG.

[0048] In some embodiments of the present invention, the C-type crystals of the compound represented by formula (I) may exist in the form of a solvate crystal.

[0049] The present invention further provides a method for producing type C crystals of the compound represented by formula (I), which comprises a step of stirring the compound represented by formula (I) in a mixed solvent of ethyl acetate / n-heptane at room temperature, and further comprises a separation step.

[0050] In some embodiments of the present invention, the C-type crystal of the compound represented by formula (I) of the present invention is (1) A method in which a mixed solvent of ethyl acetate / n-heptane is added to the compound represented by formula (I) and the mixture is stirred at room temperature; (2) Evaporation at low temperature and concentration under reduced pressure Manufactured by.

[0051] In some embodiments of the present invention, the solvent ratio of ethyl acetate / n-heptane in the above step (1) is preferably 1 / 4 (volume ratio).

[0052] In some embodiments of the present invention, the low temperature in step (2) is preferably -20°C.

[0053] The present invention further provides a D-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.136±0.200°, 9.908±0.200°, 17.787±0.200°, 18.329±0.200°, and 24.298±0.200°. [ka]

[0054] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the D-type crystal of the compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 9.136±0.200°, 9.908±0.200°, 17.787±0.200°, 18.329±0.200°, and 24.298±0.200°.

[0055] In some embodiments of the present invention, the XRPD spectrum of the D-type crystal of the compound represented by formula (I) is as shown in FIG.

[0056] In some embodiments of the present invention, the peak positions, interplanar distances and relative intensities of diffraction peaks in the powder X-ray diffraction spectrum of the D-type crystals of the compound represented by formula (I) are as shown in Table 4. [Table 4]

[0057] In some embodiments of the present invention, the differential scanning calorimetry curve of the D-type crystals of the compound represented by formula (I) has an endothermic peak value at 113.3°C ± 3°C.

[0058] In some embodiments of the present invention, the DSC spectrum of the D-type crystal of the compound represented by formula (I) is as shown in FIG.

[0059] In some embodiments of the present invention, the thermogravimetric analysis curve of the D-type crystal of the compound represented by formula (I) shows a weight loss of 6.59% at 90.0°C±3°C and a weight loss of 15.60% at 150.0°C±3°C.

[0060] In some embodiments of the present invention, the TGA spectrum of the D-type crystal of the compound represented by formula (I) is as shown in FIG.

[0061] In some embodiments of the present invention, the D-type crystals of the compound represented by formula (I) may exist in the form of a solvate crystal.

[0062] The present invention further provides a method for producing type D crystals of the compound represented by formula (I), which comprises stirring the compound represented by formula (I) in a mixed solvent of dichloromethane / n-heptane at room temperature, and further comprises a separation step.

[0063] In some embodiments of the present invention, the D-type crystal of the compound represented by formula (I) of the present invention is (1) A method in which a compound represented by formula (I) is added to a dichloromethane / n-heptane mixed solvent and stirred at room temperature; (2) Evaporation at low temperature and concentration under reduced pressure Manufactured by.

[0064] In some embodiments of the present invention, the solvent ratio of dichloromethane / n-heptane in the above step (1) is preferably 1 / 4 (volume ratio).

[0065] In some embodiments of the present invention, the low temperature in step (2) is preferably -20°C.

[0066] The present invention further provides type E crystals of the compound represented by formula (I), characterized in that the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 5.987±0.200°, 9.928±0.200°, 11.998±0.200°, 13.499±0.200°, and 18.044±0.200°. [ka]

[0067] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the E-type crystalline compound represented by formula (I) has characteristic diffraction peaks at the following 2θ angles: 5.987±0.200°, 9.928±0.200°, 11.998±0.200°, 13.499±0.200°, 18.044±0.200°, 19.492±0.200°, and 25.173±0.200°.

[0068] In some embodiments of the present invention, the XRPD spectrum of the E-form crystal of the compound represented by formula (I) is as shown in FIG.

[0069] In some embodiments of the present invention, the peak positions, interplanar distances and relative intensities of diffraction peaks in the powder X-ray diffraction spectrum of type E crystals of the compound represented by formula (I) are as shown in Table 5. [Table 5]

[0070] In some embodiments of the present invention, the differential scanning calorimetry curve of the E-form crystals of the compound represented by formula (I) has an endothermic peak value at 113.7°C±3°C.

[0071] In some embodiments of the present invention, the DSC spectrum of type E crystals of the compound represented by formula (I) is as shown in FIG.

[0072] In some embodiments of the present invention, the thermogravimetric analysis curve of the E-form crystals of the compound represented by formula (I) shows a weight loss of 13.97% at 150.0°C±3°C.

[0073] In some embodiments of the present invention, the TGA spectrum of type E crystals of the compound represented by formula (I) is as shown in FIG.

[0074] In some embodiments of the present invention, the TGA spectrum of type E crystals of the compound represented by formula (I) is as shown in FIG.

[0075] The present invention further provides a method for producing type E crystals of the compound represented by formula (I), which comprises a step of stirring the compound represented by formula (I) in toluene at room temperature, and further comprises a step of separating the compound.

[0076] In some embodiments of the present invention, the E-form crystal of the compound represented by formula (I) of the present invention is (1) A method in which toluene is added to the compound represented by formula (I) and the mixture is stirred at room temperature; (2) Evaporation and vacuum concentration Manufactured by.

[0077] In some embodiments of the present invention, the volatilization temperature in step (2) above is preferably room temperature.

[0078] The present invention further provides a crystalline composition comprising Type A crystals, Type B crystals, Type C crystals, Type D crystals, or Type E crystals of a compound of formula (I), wherein the crystals account for more than 50% by weight of the crystalline composition, preferably more than 80%, more preferably more than 90%, and most preferably more than 95%.

[0079] The present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of Type A crystals, Type B crystals, Type C crystals, Type D crystals, or Type E crystals of a compound represented by formula (I), or any of the above crystalline compositions. The pharmaceutical composition of the present invention may or may not contain a pharmaceutically acceptable excipient. Furthermore, the pharmaceutical composition of the present invention may further comprise one or more other therapeutic agents.

[0080] The present invention further provides a method for treating a coronavirus infection, comprising administering a therapeutically effective amount of Type A crystals, Type B crystals, Type C crystals, Type D crystals, or Type E crystals of the compound represented by formula (I) of the present invention, or any of the above crystalline compositions, or any of the above pharmaceutical compositions, to an individual in need thereof. The present invention further provides use of Type A crystals, Type B crystals, Type C crystals, Type D crystals, or Type E crystals of the compound represented by formula (I), or the crystalline composition, or the pharmaceutical composition, in the manufacture of a medicament for the treatment of a disease associated with a coronavirus infection.

[0081] In some embodiments of the present invention, the disease associated with coronavirus infection is SARS-CoV-2 viral infection.

[0082] [Technical Effects] The crystals of the compound of the present invention have good PK properties and therapeutic effects on SARS-CoV-2 virus, and the crystals are stable, have good hygroscopicity, and are not easily affected by light or heat.

[0083] [Definitions and Explanations] Unless otherwise stated, the following terms and phrases used herein have the following meanings. Unless otherwise defined, a particular phrase or term should be understood to have its ordinary definition, rather than being indefinite or unclear. When a trade name appears in this specification, it refers to the corresponding product or its active ingredient.

[0084] It is well known in the field of crystallography that for any given crystalline form, the relative intensities of diffraction peaks can vary due to factors such as preferred orientation due to crystal morphology. While the influence of preferred orientation can alter peak intensities, the positions of the diffraction peaks of a crystal cannot. Furthermore, it is well known in the field of crystallography that slight errors in peak positions can exist for any given crystal. For example, peak positions can shift due to temperature changes during sample analysis, sample movement, or instrument calibration, resulting in a measurement error of approximately ±0.2 degrees for the 2θ values. Therefore, it is well known to those skilled in the art that this error must be taken into account when determining the structure of each crystal.

[0085] DSC measures the transition temperatures that occur when heat is absorbed or released due to changes in crystalline structure or crystalline melting. For the same crystal of the same compound, the error between the thermal transition temperature and melting point in consecutive analyses is usually within approximately 5°C or 3°C. If a compound has a specific DSC peak or melting point, this means that the DSC peak or melting point is within ±5°C or ±3°C. DSC provides an auxiliary method for distinguishing different crystals. Different crystals can be identified based on their different transition temperature characteristics. It should be noted that for mixtures, the DSC peak or melting point may vary over a wider range. Furthermore, because the melting process of a substance involves decomposition, the melting temperature is related to the heating rate.

[0086] For the same crystal, the weight loss temperature in TGA may vary depending on the measuring instrument, measurement method / conditions, etc. For any particular crystal, there may be an error in the weight loss temperature, and the error may be about ±5°C or about ±3°C.

[0087] It should be noted that in the preparation of drug crystals, when drug molecules and solvent molecules come into contact with each other, external and internal conditions can cause the solvent molecules and compound molecules to form a eutectic, which is difficult to avoid remaining in the solid material, resulting in the formation of solvates, specifically including stoichiometric solvates and non-stoichiometric solvates. Both of these solvates are included within the scope of the present invention.

[0088] The term "pharmaceutically acceptable excipient" refers to an inert substance that is co-administered with an active ingredient to facilitate administration of the active ingredient, and includes, but is not limited to, any flow aid, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that is approved by the Food and Drug Administration for use in humans or animals (e.g., livestock).

[0089] The term "crystalline composition" refers to a mixture of a crystal of the compound of formula (I) of the present invention with other crystalline or amorphous substances of the compound or other impurities. For example, a crystalline composition of type A crystal of the compound of formula (I) contains, in addition to type A crystal of the compound of formula (I), other crystalline or amorphous substances of the compound of formula (I), or other impurities.

[0090] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or salts thereof with a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present invention to an organism.

[0091] The therapeutic dosage of a compound of the invention may be determined based on, for example, the particular therapeutic application, the manner in which the compound is administered, the patient's health condition, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition may not be fixed and will depend upon various factors, including dosage, chemical characteristics (e.g., hydrophobicity), and the route of administration.

[0092] The term "treatment" means the administration of a compound or formulation according to the present invention to ameliorate or eliminate a disease or one or more symptoms associated with said disease, and includes: (i) inhibiting a disease or disease state, i.e., preventing its onset; (ii) Alleviation of the disease or disease state, i.e., resolution of the disease or disease state.

[0093] The term "therapeutically effective amount" refers to an amount of a compound of the present invention that (i) treats a particular disease, condition, or disorder, (ii) reduces, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the method of administration, and the age of the mammal being treated, but can be determined routinely by one of ordinary skill in the art in light of their own knowledge and the present disclosure.

[0094] Unless otherwise required by the present invention, throughout this specification and the claims that follow, the word "comprise" and its English variants such as "comprises" and "comprising" are to be interpreted in an open-ended and inclusive sense, i.e., "including but not limited to."

[0095] Throughout this specification, a reference to "one embodiment" or "an embodiment" or "another embodiment" or "in some embodiments" means that at least one embodiment includes the particular referenced element, structure, or feature described in that embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" or "in another embodiment" or "in some embodiments" in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, particular elements, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0096] As used in this specification and the appended claims, the singular form "a" (corresponding to the English words "a," "an," and "the") should be understood to include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a reaction including a "catalyst" includes one catalyst, or two or more catalysts. It should further be understood that the term "or" is generally used in its sense to include "and / or" unless the context clearly dictates otherwise.

[0097] The intermediate compounds of the present invention can be prepared by various synthetic methods familiar to those skilled in the art, including the specific embodiments listed below, embodiments in combination with other chemical synthetic methods, and equivalent alternative methods familiar to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0098] The chemical reactions of specific embodiments of the present invention are completed in suitable solvents, which should be suitable for the chemical reactions of the present invention and the reagents and materials required therefor. To obtain the compounds of the present invention, those skilled in the art may need to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0099] The present invention will be specifically described below with reference to examples, but these examples do not limit the present invention in any way.

[0100] All solvents used in this invention are commercially available and may be used as is without further purification.

[0101] Compounds are named according to conventional naming principles in the art or in ChemDraw (R) Compounds are named using the software, and commercially available compounds are named in the supplier's catalog.

[0102] Instrumentation and analytical methods

[0103] 1.1 X-ray powder diffractometer (XRPD) method of the present invention Instrument model: PANalytacal X-ray diffractometer Test method: Approximately 1-2 mg of sample is used for XRPD detection. The detailed XRPD parameters are as shown below. X-ray tube: Cu, kα, Kα1(Å): 1.540598, Kα2(Å): 1.544426, Kα2 / Kα1 intensity ratio: 0.50 Tube voltage: 45 kV, tube current: 40 mA Divergence slit: 1 / 8° Scanning mode: Continuous Scanning range (°2 Theta): 3~40 Scan time per step (s): 46.7 Scan step width (°2 Theta): 0.0263 Test time: ~5 minutes

[0104] 1.2 Differential Scanning Calorimeter (DSC) Method of the Present Invention Instrument model: TA 2500 Differential Scanning Calorimeter Measurement method: A sample (up to 1 mg) is placed in a DSC aluminum crucible and heated from 25°C (room temperature) to the installation temperature at a rate of 10°C / min under conditions of 50 mL / min N2.

[0105] 1.3 Thermogravimetric Analysis (TGA) Method of the Present Invention Instrument model: TA 5500 Thermogravimetric Analyzer Measurement method: A sample (up to 1 mg) is placed in a TGA platinum crucible and heated from room temperature to the installation temperature at a rate of 10°C / min under conditions of 25 mL / min N2. or: Thermogravimetric analyzer TGA550 Measurement method: A sample (5-10 mg) is placed on the built-in aluminum plate of a TGA platinum crucible and heated from room temperature to 300°C at a heating rate of 10°C / min under conditions of 60 mL / min N2.

[0106] 1.4 Dynamic Vapor Sorption (DVS) Method of the Present Invention Instrument Model: SMS DVS Advantage Dynamic Vapor Sorption Apparatus Measurement conditions: A sample (10 to 30 mg) was placed in a DVS sample pan and the test was carried out. The detailed DVS parameters are as follows: Temperature: 25℃ Balance: dm / dt = 0.002% / min (min: 10 min, max: 180 min) RH (%) range: 70%RH~95%RH~0%RH~95%RH RH (%) gradient: 10% (90%RH~0%RH~90%RH), 5% (95%RH~90%RH and 90%RH~95%RH). [Brief explanation of the drawings]

[0107] [Figure 1] 1 is an XRPD spectrum of Cu-Kα radiation of type A crystal of the compound represented by formula (I). [Figure 2] 1 is a DSC spectrum of type A crystals of the compound represented by formula (I). [Figure 3] 1 is a TGA spectrum of type A crystals of the compound represented by formula (I). [Figure 4] 1 is an XRPD spectrum of Cu-Kα radiation of type B crystals of the compound represented by formula (I). [Figure 5] 1 is a DSC spectrum of type B crystals of the compound represented by formula (I). [Figure 6] 1 is a TGA spectrum of type B crystals of the compound represented by formula (I). [Figure 7] 1 is an XRPD spectrum of Cu-Kα radiation of a C-type crystal of the compound represented by formula (I). [Figure 8] 1 is a DSC spectrum of type C crystals of the compound represented by formula (I). [Figure 9] 1 is a TGA spectrum of type C crystals of the compound represented by formula (I). [Figure 10] 1 is an XRPD spectrum of Cu-Kα radiation of a D-type crystal of the compound represented by formula (I). [Figure 11] 1 is a DSC spectrum of type D crystals of the compound represented by formula (I). [Figure 12] 1 is a TGA spectrum of type D crystals of the compound represented by formula (I). [Figure 13] 1 is an XRPD spectrum of Cu-Kα radiation of E-form crystals of the compound represented by formula (I). [Figure 14] 1 is a DSC spectrum of type E crystals of the compound represented by formula (I). [Figure 15] 1 is a TGA spectrum of type E crystals of the compound represented by formula (I). [Figure 16] 1 is a DVS spectrum of type A crystal of the compound represented by formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0108] The present invention will be specifically described below by way of examples, but is not intended to be an adverse limitation of the present invention. The present invention has been described in detail herein, and specific embodiments thereof have been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made in the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0109] Example 1: Preparation of Compound 1 [ka]

[0110] Step 1: Synthesis of Compound 1-2 Compound 1-1 (5 g, 54.32 mmol) was dissolved in methanol (50 mL) and refluxed at 70 °C for 48 hours. The reaction mixture was concentrated under reduced pressure to give the crude target product. The crude product was highly pure and was directly used in the next step to give compound 1-2. 1 H NMR (400 MHz, CDCl3) δ = 4.81 (s, 1H), 3.77 (s, 3H), 3.43 (s, 3H).

[0111] Step 2: Synthesis of Compounds 1-3 Compound 1-2 was dissolved in toluene (3 mL) and cooled to 0 °C. Compound (R)-(+)-phenethylamine (1.5 g, 12.38 mmol, 1.60 mL) was slowly added dropwise and stirred at 20 °C for 1 h. The reaction mixture was extracted with ethyl acetate (60 mL) and saturated brine (30 mL). The organic phase was separated, dried over anhydrous sodium sulfate, and spin-dried to obtain the crude product. The product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 5:1) to obtain the target compound 1-3. 1 H NMR (400 MHz, CDCl3) δ = 7.95~7.56 (m, 1H), 7.31~7.17 (m, 5H), 4.71~4.40 (m, 1H), 3.95~3.71 (m, 3H), 1.67~1.51 (m, 3H).

[0112] Step 3: Synthesis of Compounds 1-4 Compound 1-3 (0.5 g, 2.61 mmol) was dissolved in 2,2,2-trifluoroethanol (5 mL), trifluoroacetic acid (313.04 mg, 2.75 mmol, 203.28 μL) was added, and the mixture was cooled to -10 °C and stirred for 1 h. The temperature was controlled at -10 °C, and cyclopentadiene (207.40 mg, 3.14 mmol) was slowly added dropwise, followed by stirring for 0.5 h. The reaction mixture was concentrated under reduced pressure, and methyl tert-butyl ether (60 mL) and saturated sodium bicarbonate solution (30 mL × 2) were added. The mixture was stirred for 10 min, extracted, and the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Compound 1-4 was obtained by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 5:1). The stereochemistry was confirmed by 2D NMR. 1H NMR (400 MHz, CDCl3) δ = 7.34~7.18 (m, 5H), 6.59~6.41 (m, 1H), 6.31 (dd, J = 1.6, 5.6 Hz, 1H), 4.35 (br d, J = 1.3 Hz, 1H), 3.39 (s, 3H), 3.18~3.03 (m, 1H), 2.95 (br s, 1H), 2.33~2.22 (m, 1H), 2.14 (br d, J = 8.4 Hz, 1H), 1.54~1.41 (m, 4H), MS m / z(ESI):[M+H] + =258.2.

[0113] Step 4: Synthesis of Compounds 1-5 Compound 1-4 (100.00 mg, 388.61 μmol) was dissolved in tetrahydrofuran (1.25 mL) and cooled to -70 °C. Borane tetrahydrofuran complex (1 M, 427.47 μL) was slowly added dropwise, and the mixture was slowly warmed to 20 °C and stirred for 1 hour. After cooling to 0 °C, a 10% aqueous sodium hydroxide solution (0.55 mL) and a 30% aqueous hydrogen peroxide solution (220.28 mg, 1.94 mmol, 186.68 μL) were added, and the mixture was slowly warmed to 20 °C and stirred for 1 hour. A saturated aqueous sodium thiosulfate solution (10 mL) was added to the reaction mixture and stirred for 10 minutes to quench the reaction. The mixture was then extracted with saturated brine (20 mL) and ethyl acetate (60 mL × 2), and the organic phase was separated. A small amount of the sample solution was taken, and the pH was adjusted to less than 8 with 3% citric acid. After the starch potassium iodide test paper showed a negative result, the solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 30°C. Compound 1-5 was obtained by purification using silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 5:1). 1H NMR (400 MHz, CDCl3) δ = 7.30~7.13 (m, 5H), 3.93 (br d, J = 6.5 Hz, 1H), 3.78 (br s, 1H), 3.70~3.54 (m, 1H), 3.39~3.32 (m, 1H), 3.31~3.24 (m, 3H), 2.49~2.40 (m, 1H), 2.26 (s, 1H), 2.09~2.00 (m, 1H), 1.72 (br d, J = 10.1 Hz, 1H), 1.46 (br d, J = 6.5 Hz, 1H), 1.41~1.33 (m, 3H), MS m / z(ESI):[M+H] + =276.1.

[0114] Step 5: Synthesis of the hydrochloride salts of compounds 1-6 Compound 1-5 (3 g, 10.90 mmol) was dissolved in ethanol (80 mL), and hydrochloric acid (1.19 g, 32.69 mmol) and wet palladium on carbon (15 g, 10.68 mmol) were added. The reaction was stirred at 20 °C for 16 h. The reaction mixture was filtered through diatomaceous earth and then directly spun dry to give the crude product, compound 1-6, hydrochloride salt. 1 H NMR (400 MHz, DMSO-d6) δ = 10.32~9.78 (m, 1H), 8.94~8.43 (m, 1H), 5.51~5.12 (m, 1H), 4.05~3.98 (m, 1H), 3.96~3.86 (m, 2H), 3.83~3.71 (m, 3H), 2.70~2.60 (m, 1H), 2.36~2.20 (m, 1H), 1.92~1.81 (m, 1H), 1.51~1.31 (m, 2H), MS m / z(ESI):[M+H] + =172.0.

[0115] Step 6: Synthesis of Compounds 1-8 Compound 1-7 (1.87 g, 10.90 mmol) was dissolved in N,N-dimethylformamide (20 mL), and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (4.78 g, 12.58 mmol) and diisopropylethylamine (4.34 g, 33.55 mmol) were added. After stirring for 30 minutes, compound 1-6 hydrochloride (190 mg, 1.12 mmol) was added. The reaction mixture was stirred at 20 °C for 16 hours. Water (15 mL) was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate (60 mL). The combined organic phase was washed twice with 5% citric acid (30 mL), washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and spun dry. Purification by column chromatography (petroleum ether:ethyl acetate=3:1) gave compound 1-8. 1 H NMR (400 MHz, CDCl3) δ = 5.28~5.16 (m, 1H), 4.50 (br s, 1H), 4.28 (d, J = 9.8 Hz, 1H), 3.92 (s, 1H), 3.74 (s, 3H), 2.81 (s, 1H), 2.67 (s, 1H), 2.17 (br dd, J = 6.1, 12.7 Hz, 1H), 1.99~1.93 (m, 1H), 1.90~1.84 (m, 1H), 1.59 (br d, J = 13.3 Hz, 2H), 1.43 (s, 9H), 1.04 (s, 9H), MS m / z(ESI):[M+H] + =385.2.

[0116] Step 7: Synthesis of Compounds 1-9 Compound 1-8 (500 mg, 1.30 mmol) was dissolved in acetonitrile (7.5 mL), and 2-iodoxybenzoic acid (976.31 mg, 3.49 mmol) was added. The mixture was stirred at 60° C. for 16 hours. The reaction mixture was directly filtered through diatomaceous earth and spin-dried. Compound 1-9 was obtained without further purification. MS m / z(ESI):[M-55] + =327.1.

[0117] Step 8: Synthesis of Compounds 1-10 Compound 1-9 (0.7 g, 1.83 mmol) was dissolved in tetrahydrofuran (14 mL) and TEBBE (μ-chloro-μ-methylene[bis(cyclopentadienyl)titanium]dimethylaluminum) reagent (0.5 M, 14.64 mL) was added at 0 °C. The mixture was stirred at 0 °C for 1 hour, then warmed to 15 °C and stirred for 3 hours. The reaction mixture was slowly poured into saturated sodium bicarbonate solution (50 mL), filtered through diatomaceous earth, extracted with ethyl acetate (30 mL × 3), and washed with saturated brine (30 mL × 2). Compound 1-10 was obtained by column chromatography (petroleum ether:ethyl acetate = 5:1). 1 H NMR (400 MHz, DMSO-d6) δ = 6.63~6.54 (m, 1H), 5.21~5.15 (m, 1H), 4.88~4.82 (m, 1H), 4.76~4.68 (m, 1H), 4.25~4.19 (m, 1H), 3.90~3.85 (m, 1H), 3.66~3.61 (m, 3H), 3.19~3.11 (m, 1H), 2.42~2.28 (m, 2H), 1.98~1.92 (m, 1H), 1.61~1.53 (m, 1H), 1.38 (s, 9H), 1.00~0.93 (m, 9H), MS m / z(ESI):[M+H] + =381.1.

[0118] Step 9: Synthesis of Compounds 1-11 Under nitrogen gas protection, diethylzinc (1M, 13.14 mL) was slowly added to 1,2-dichloroethane (80 mL) at 0 °C. After stirring for 0.25 h, diiodomethane (7.04 g, 26.28 mmol, 2.12 mL) was slowly added to the reaction mixture at 0 °C and stirred for 0.25 h. Trifluoroacetic acid (149.84 mg, 1.31 mmol, 97.30 μL) was slowly added to the reaction mixture and stirring was continued for 0.5 h. Compound 1-10 (0.5 g, 1.31 mmol) in 1,2-dichloroethane (5 mL) was added to the reaction mixture, and the mixture was warmed to 20 °C and stirred for 12 h. The reaction mixture was quenched with saturated sodium bicarbonate solution (200 mL), extracted with dichloromethane (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative HPLC (column type: Phenomenex Luna C18 80 × 0 mm × 3 μm; mobile phase: [HO(HCl)-acetonitrile]; acetonitrile %: 1% to 30%, 7 min) to give compound 1-11. MS m / z (ESI): [M+H] + =295.2.

[0119] Step 10: Synthesis of Compounds 1-12 Compound 1-11 (0.1 g, 339.69 μmol) was dissolved in 1,4-dioxane (3 mL). Then, a solution of potassium carbonate (187.79 mg, 1.36 mmol) and di-tert-butyl dicarbonate (111.20 mg, 509.53 μmol, 117.06 μL) in water (1 mL) was added, and the reaction mixture was stirred at 15 °C for 12 hours. The reaction mixture was poured into water (30 mL), extracted with ethyl acetate (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by column chromatography (petroleum ether:ethyl acetate = 5:1) gave compound 1-12. 1H NMR (400 MHz, DMSO-d6) δ = 12.52~12.16 (m, 1H), 6.56~6.31 (m, 1H), 4.66~4.58 (m, 1H), 4.24~4.18 (m, 1H), 4.02 (s, 1H), 3.36~3.28 (m, 1H), 1.97~1.91 (m, 2H), 1.81~1.73 (m, 2H), 1.66~1.59 (m, 1H), 1.36 (s, 9H), 0.99~0.93 (m, 9H), 0.80~0.70 (m, 1H), 0.64~0.53 (m, 1H), 0.49~0.33 (m, 2H) MS m / z(ESI):[M+H] + =395.2.

[0120] Step 11: Synthesis of Compounds 1-13 Compound 1-12 (88.13 mg, 223.40 μmol) was dissolved in tetrahydrofuran (2 mL) and methanol (0.6 mL), and lithium hydroxide monohydrate (28.12 mg, 670.21 μmol) dissolved in water (0.6 mL) was added. The reaction mixture was stirred at 15° C. for 2 hours. The pH of the reaction mixture was adjusted to approximately 5 with 3% citric acid, extracted with ethyl acetate (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1-13. MS m / z(ESI):[M+H] + =381.3.

[0121] Step 12: Synthesis of Compounds 1-15 Compound 1-13 (0.056 g, 148.79 μmol) was dissolved in N,N-dimethylformamide (2 mL), and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (84.86 mg, 223.18 μmol) was added to the reaction mixture, which was then stirred at 15° C. for 0.5 hours. Next, diisopropylethylamine (76.92 mg, 595.16 μmol, 103.67 μL) was added to the reaction mixture, and the hydrochloride salt of compound 1-14 (43.26 mg, 208.31 μmol) was dissolved in N,N-dimethylformamide (0.5 mL) and added to the reaction mixture. The reaction mixture was then stirred at 15° C. for 12 hours. The reaction was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3), and the organic phase was washed with 3% citric acid (20 mL), washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1-15. 1 H NMR (400 MHz, DMSO-d6) δ = 8.22~8.08 (m, 1H), 7.56 (s, 1H), 7.32~7.16 (m, 1H), 7.01 (s, 1H), 6.51 (br d, J = 9.4 Hz, 1H), 4.60~4.49 (m, 1H), 4.28~4.17 (m, 2H), 4.14 (s, 1H), 3.18~2.98 (m, 2H), 2.47~2.35 (m, 1H), 2.17~2.09 (m, 2H), 1.96~1.84 (m, 2H), 1.76 (br d, J = 11.0 Hz, 1H), 1.71~1.42 (m, 4H), 1.39~1.34 (m, 9H), 0.95 (s, 8H), 0.84~0.77 (m, 1H), 0.73~0.65 (m, 1H), 0.39 (br s, 2H), MS m / z(ESI):[M+H] + =534.4.

[0122] Step 13: Synthesis of Compounds 1-16 Compound 1-15 (0.02 g, 37.48 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (141.02 mg, 1.24 mmol, 91.57 μL) was added to the reaction mixture. The mixture was stirred at 15 °C for 1 hour. The reaction mixture was directly quenched with sodium bicarbonate solution (10 mL) and extracted with dichloromethane (5 mL × 5). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 1-16. 1 H NMR (400 MHz, DMSO-d6) δ = 8.37~8.26 (m, 1H), 8.14 (br d, J = 3.6 Hz, 3H), 7.59~7.53 (m, 1H), 7.40~7.32 (m, 1H), 7.05~6.92 (m, 1H), 4.33~4.21 (m, 2H), 3.93 (br d, J = 5.0 Hz, 1H), 3.17~3.08 (m, 1H), 3.06~2.97 (m, 1H), 2.44~2.32 (m, 1H), 2.18~2.08 (m, 2H), 1.96~1.86 (m, 2H), 1.82~1.57 (m, 4H), 1.51~1.40 (m, 1H), 1.04 (s, 9H), 0.95~0.85 (m, 1H), 0.76~0.67 (m, 1H), 0.54~0.34 (m, 2H), MS m / z(ESI):[M+H] + =434.2.

[0123] Step 14: Synthesis of Compound 1 Compound 1-16 (0.03 g, 69.20 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic anhydride (58.13 mg, 276.79 μmol, 38.50 μL) was added to the reaction mixture. The mixture was stirred at 15 °C for 1 h. The reaction mixture was directly quenched with sodium bicarbonate solution (10 mL) and extracted with dichloromethane (5 mL × 5). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by preparative HPLC (column type: Waters Xbridge BEH C18 100 × 30 mm × 10 μm; mobile phase: [HO(NHHCO)-acetonitrile]; acetonitrile %: 10% to 50%, 8 min) to obtain compound 1, i.e., the compound represented by formula (I). 1 H NMR (400 MHz, DMSO-d6) δ = 9.31 (br d, J = 8.0 Hz, 1H), 8.87 (d, J = 8.6 Hz, 1H), 7.66 (s, 1H), 5.01~4.87 (m, 1H), 4.72~4.59 (m, 2H), 4.07 (s, 1H), 3.17~3.10 (m, 1H), 3.09~2.98 (m, 1H), 2.45~2.33 (m, 1H), 2.19~2.05 (m, 3H), 1.85~1.64 (m, 5H), 1.56 (br d, J = 12.0Hz, 1H), 1.20~1.13 (m, 1H), 1.01 (s, 8H), 0.82~0.67 (m, 2H), 0.54~0.46 (m, 1H), 0.45~0.34 (m, 1H), MS m / z(ESI):[M+H] + =512.2.

[0124] Example 2: Preparation of Type A Crystals of the Compound of Formula (I) The compound of formula (I) (9.4 g, 18.38 mmol) was dissolved in HO (235 mL) and stirred at 50° C. for 48 hours. The resulting solid was filtered to give a white solid, which was then concentrated under reduced pressure to remove water, yielding Type A crystals, which were detected by XRPD.

[0125] Example 3: Preparation of B-type crystals of the compound represented by formula (I) The compound represented by formula (I) (15 mg, 29.32 μmol) was placed in a glass vial, and 0.5 mL of ethanol / n-heptane (1:4) solvent was added. The sample was placed at room temperature and magnetically stirred (1000 rpm) for about 5 days. The sample was then cooled to a low temperature (5°C) while stirring to evaporate the solid. The solid was collected by centrifugation and concentrated under reduced pressure to obtain type B crystals, which were detected by XRPD.

[0126] Example 4: Preparation of C-type crystals of the compound represented by formula (I) The compound represented by formula (I) (15 mg, 29.32 μmol) was placed in a glass vial, and 0.5 mL of ethyl acetate / n-heptane (1:4) solvent was added. The sample was placed at room temperature and magnetically stirred (1000 rpm) for about 5 days. The sample was then cooled to a low temperature (−20°C) with stirring to evaporate the solid. The solid was collected by centrifugation and concentrated under reduced pressure to obtain type C crystals, which were detected by XRPD.

[0127] Example 5: Preparation of D-type crystals of the compound represented by formula (I) The compound represented by formula (I) (15 mg, 29.32 μmol) was placed in a glass vial, and 0.5 mL of dichloromethane / n-heptane (1:4) solvent was added. The sample was placed at room temperature and magnetically stirred (1000 rpm) for about 5 days. The sample was then cooled to a low temperature (−20°C) with stirring to evaporate the solid. The solid was collected by centrifugation and concentrated under reduced pressure to obtain type D crystals, which were detected by XRPD.

[0128] Example 6: Preparation of E-type crystals of the compound represented by formula (I) The compound represented by formula (I) (15 mg, 29.32 μmol) was placed in a glass vial, and 0.5 mL of toluene solvent was added. The sample was left at room temperature until it dissolved into a transparent state, and then magnetically stirred (1000 rpm) for about 5 days. Next, the sample was cooled to a low temperature (-20°C) while stirring, but no precipitation was observed. The sample was then heated to room temperature and evaporated to obtain a white solid. The solid was concentrated under reduced pressure to obtain E-type crystals, which were detected by XRPD.

[0129] Example 7: Hygroscopicity study of crystalline form A of compound of formula (I) Test materials: SMS DVS Advantage Dynamic Vapor Sorption Apparatus Experimental Method: 10 to 30 mg of the type A crystals of the compound represented by formula (I) were placed in a DVS sample pan and measured. Test Results: The DVS spectrum of the type A crystal of the compound represented by formula (I) was shown in FIG. 16, and ΔW was 0.114%. Testing Conclusion: The A-type crystals of the compound represented by formula (I) showed a hygroscopic weight gain of 0.114% at 25°C and 80% RH, and were either not hygroscopic or had little hygroscopicity.

[0130] Example 8: Stability study of crystalline form A of compound of formula (I) The results of stability tests of Form A crystals under different conditions are shown in Table 6. [Table 6] Experimental conclusion: The A-type crystal of the compound of the present invention has good stability under conditions of light irradiation, high temperature and high humidity.

[0131] Biological Test Data:

[0132] Experimental Example 1: Evaluation of in vitro anti-coronavirus Mpro protease activity of test compounds 1. Experimental materials: 1.1 Reagents and consumables are as shown in Table 7. [Table 7]

[0133] 1.2 The equipment and brands are as shown in Table 8. [Table 8]

[0134] 2. Experimental Method: Compounds were dissolved in DMSO and diluted threefold using an Echo 655 to create 10 concentration points, each with two replicate wells, into a 384-well plate. Mpro protein and substrate were diluted in test buffer (100 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA). Mpro protein was added to the 384-well plate and incubated with the compound at room temperature for 30 minutes. Substrate was then added, with the Mpro protein test concentration at 25 nM and the substrate test concentration at 25 μM. The plates were then incubated in a 30°C incubator for 60 minutes. The fluorescence signal was then measured using a microplate reader at Ex / Em = 340 nm / 490 nm. Simultaneously, background wells containing the substrate and compound but no Mpro protein were used as controls.

[0135] 3. Data Analysis: 1) The inhibition rate was calculated using the following formula: Inhibition rate % = [(compound - BG 化合物 )-(ZPE-BG ZPE )] / [(HPE-BG HPE )-(ZPE-BG ZPE )] × 100% # HPE: 100% inhibition control, containing 25 nM Mpro protein + 25 μM substrate + 1 μM GC376 ZPE: No inhibition control, 25 nM Mpro protein + 25 μM substrate, no compound Compound: Test compound wells, containing 25 nM Mpro protein + 25 μM substrate + compound BG: background control well, containing 25 μM substrate + compound, no Mpro protein 2) Using GraphPad Prism software, perform a nonlinear fitting analysis of the log(agonist) vs. response variable slope for the compound inhibition rate data (% inhibition) to obtain the compound's IC 50 got the value. [Table 9] Conclusion: The compounds of the present invention have good in vitro anti-coronavirus Mpro protease activity.

[0136] Example 2: Evaluation of in vitro anti-coronavirus activity of compounds using a cytopathic model 1. The experimental materials are as shown in Tables 10 and 11. [Table 10] [Table 11]

[0137] 1.1 Cells and viruses MRC5 cells and coronavirus HCoV OC43 were purchased from ATCC. MRC5 cells were cultured in MEM (Sigma) medium supplemented with 10% fetal bovine serum (Excell), 1% double antibody (Hyclone), 1% L-glutamine (Gibco), and 1% non-essential amino acids (Gibco). Experimental medium was MEM (Sigma) medium supplemented with 5% fetal bovine serum (Excell), 1% double antibody (Hyclone), 1% L-glutamine (Gibco), and 1% non-essential amino acids (Gibco).

[0138] 2. Experimental Method [Table 12] Cells were seeded into 96-well microplates at a predetermined density (Table 12) and cultured overnight in a 5% CO2, 37°C incubator. The next day, diluted compounds (8 concentration points, 2 replicate wells) were added at 50 μL per well. Diluted virus was then added at 100 TCID per well. 50The cells were added at 50 μL at 100°C. A cell control (cells without compound treatment or virus infection), a virus control (virus-infected cells without compound treatment), and a medium control (medium only) were set up. The final volume of the experimental medium was 200 μL, and the final DMSO concentration in the medium was 0.5%. The cells were cultured in a 5% CO2, 33°C incubator for 5 days. Cell viability was detected using the CellTiter Glo (Promega) cell activity detection kit. The conditions for the cytotoxicity experiment were the same as those for the antiviral experiment, except that there was no virus infection.

[0139] 3. Data Analysis The antiviral activity and cytotoxicity of the compounds were expressed as the inhibition rate (%) of the cytopathic effect caused by the virus and the cell viability (%) at different concentrations of the compounds, respectively. The calculation formulas are as follows: Inhibition rate (%) = (test well reading - mean value of virus control) / (mean value of cell control - mean value of virus control) x 100 Cell viability (%) = (test well reading - mean of medium control) / (mean of cell control - mean of medium control) x 100 GraphPad Prism was used to perform nonlinear fitting analysis of the compound inhibition rate and cell viability, and the median effective concentration (EC 50 ) and semi-cytotoxic concentration (CC 50 ) values ​​were calculated. [Table 13] Conclusion: The compounds of the present invention have good in vitro anti-coronavirus activity at the cellular level and no cytotoxicity.

[0140] Experimental Example 3: Pharmacokinetic study in rats In this study, male and female SD rats were selected as test animals, and compound 1 type A crystals were administered intravenously at a single dose of 2 mg / kg and intragastrically at a single dose of 30 mg / kg. The plasma drug concentrations of the test compound at different time points were then quantitatively measured using LC / MS / MS to evaluate the pharmacokinetic properties of the test drug in the rat body. The animals were divided into groups of 3 animals per sex and administered type A crystals of Compound 1. Plasma samples were collected at 0.083 (intravenous injection group only), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration, and plasma drug concentrations were measured using LC-MS / MS. The experimental results are shown in Table 14. [Table 14] Experimental Conclusion: The compounds of the present invention showed high exposure in plasma and high bioavailability.

[0141] Experimental Example 4: Evaluation of the in vivo antiviral effect of test compounds using an infant mouse infection model with the coronavirus OC43 strain C57BL / 6J infant mice were infected intranasally with a lethal dose of coronavirus and treated with vehicle (5% DMSO + 40% PEG400 + 55% water) or Compound 1 2 hours before infection. During the experiment, the weight, health, and survival of the infant mice were monitored daily to assess the protective effect of different doses of Compound 1. The weight of infant mice in the vehicle group continued to decrease on the sixth day after virus inoculation, and the endpoint survival rate was 0%. When Compound 1 (12.5, 25, 50 mpk) was first administered 2 hours before infection, the endpoint survival rates were 87.5%, 100%, and 100%, respectively. The experimental results are shown in Table 15.

[0142] [Table 15] Experimental conclusion: Compound 1 has excellent anti-coronavirus effects in vivo and shows a good dose-effect relationship.

Claims

1. A type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 10.655±0.200°, 11.988±0.200°, 16.055±0.200°, 18.356±0.200°, and 20.083±0.200°. 【Chemical 1】

2. The A-type crystal of the compound of formula (I) according to claim 1, characterized in that the A-type crystal contains at least 6, 7, or 8 diffraction peaks selected from 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, and 20.083±0.200° in a powder X-ray diffraction spectrum expressed in 2θ angles.

3. The A-type crystal of the compound represented by formula (I) according to claim 1, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, and 20.083±0.200°.

4. The A-type crystal has the following peaks in the powder X-ray diffraction spectrum represented by 2θ angles: 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 13.837±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, 20.083±0.200°, 20.839±0.200° 2. A type A crystal of the compound of formula (I) according to claim 1, characterized in that it contains at least 12, 13, 14, 15, or 16 diffraction peaks selected from 21.388±0.200°, 22.379±0.200°, 24.577±0.200°, 25.104±0.200°, 26.402±0.200°, and 31.540±0.200°.

5. The powder X-ray diffraction spectrum shows the following 2θ angles: 6.917±0.200°, 10.655±0.200°, 11.988±0.200°, 13.837±0.200°, 14.481±0.200°, 16.055±0.200°, 17.653±0.200°, 18.356±0.200°, 20.083±0.200°.

2. A type A crystal of the compound of formula (I) according to claim 1, having characteristic diffraction peaks at 0°, 20.839±0.200°, 21.388±0.200°, 22.379±0.200°, 24.577±0.200°, 25.104±0.200°, 26.402±0.200°, and 31.540±0.200°.

6. 2. The A-type crystal of the compound of formula (I) according to claim 1, characterized in that the powder X-ray diffraction spectrum of the A-type crystal is as shown in FIG.

7. 2. The A-type crystal of the compound represented by formula (I) according to claim 1, characterized in that the differential scanning calorimetry curve of the A-type crystal has an endothermic peak value at 173.5°C ± 3°C.

8. 2. A type A crystal of the compound of formula (I) according to claim 1, whose differential scanning calorimetry curve is as shown in FIG.

9. The A-type crystal of the compound of formula (I) according to claim 1, characterized in that the thermogravimetric analysis curve of the A-type crystal shows a weight loss of 0.00% at 150.0°C ± 3°C.

10. 2. A type A crystal of the compound represented by formula (I) according to claim 1, whose thermogravimetric analysis curve is as shown in FIG.

11. A B-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.202±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, and 19.917±0.200°. 【Chemistry 2】

12. The B-type crystal of the compound represented by formula (I) according to claim 11, characterized in that the B-type crystal contains at least 6, 7, or 8 diffraction peaks selected from 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, 18.306±0.200°, and 19.917±0.200° in a powder X-ray diffraction spectrum expressed in 2θ angles.

13. The B-type crystal of the compound represented by formula (I) according to claim 11, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, 18.306±0.200°, and 19.917±0.200°.

14. The B-type crystals have the following peaks in the powder X-ray diffraction spectrum expressed as 2θ angles: 5.459±0.200°, 7.096±0.200°, 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, 18.306±0.200°, 19.488±0.200°, 20.516±0.200°, 21.516±0.200°, 22.516±0.200°, 23.516±0.200°, 24.516±0.200°, 25.516±0.200°, 26.483±0.200°, 27.516±0.200°, 28.516±0.200°, 29.516±0.200°, 30.516±0.200°, 31.516±0.200°, 32.516±0.200°, 33.516±0.200°, 34.516±0.200°, 35.516±0.200°, 36.516±0.200°, 37.516±0.200°, 38.516±0.200°, 39.516±0.200°, 40.516±0.200°, 41.516±0.200°, 42.516± 12. The B-type crystal of the compound of formula (I) according to claim 11, characterized in that it contains at least 12, 13, 14, 15, or 16 diffraction peaks selected from the group consisting of 19.917±0.200°, 20.681±0.200°, 21.526±0.200°, 22.409±0.200°, 24.622±0.200°, and 25.661±0.200°.

15. The powder X-ray diffraction spectrum exhibited the following 2θ angles: 5.459±0.200°, 7.096±0.200°, 9.202±0.200°, 10.968±0.200°, 13.394±0.200°, 14.212±0.200°, 15.516±0.200°, 16.483±0.200°, and 18.306±0.200°.

12. The B-type crystal of the compound represented by formula (I) according to claim 11, having characteristic diffraction peaks at 19.488±0.200°, 19.917±0.200°, 20.681±0.200°, 21.526±0.200°, 22.409±0.200°, 24.622±0.200°, and 25.661±0.200°.

16. 12. The B-type crystal of the compound of formula (I) according to claim 11, characterized in that the powder X-ray diffraction spectrum of the B-type crystal is as shown in Figure 4.

17. 12. The B-type crystal of the compound represented by formula (I) according to claim 11, characterized in that the differential scanning calorimetry curve of the B-type crystal has an endothermic peak value at 128.4°C ± 3°C.

18. 12. The B-type crystal of the compound represented by formula (I) according to claim 11, whose differential scanning calorimetry spectrum is as shown in FIG.

19. The B-type crystal of the compound of formula (I) according to claim 11, characterized in that the thermogravimetric analysis curve of the B-type crystal shows a weight loss of 9.77% at 150.0°C ± 3°C.

20. 12. The B-type crystal of the compound represented by formula (I) according to claim 11, having a thermogravimetric analysis curve as shown in FIG.

21. The B-type crystals of the compound of formula (I) according to claim 11, wherein the B-type crystals may exist in the form of solvate crystals.

22. A C-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 6.334±0.200°, 8.399±0.200°, 9.435±0.200°, 12.060±0.200°, and 18.126±0.200°. 【Chemistry 3】

23. The C-type crystal of the compound of formula (I) according to claim 22, characterized in that the C-type crystal contains at least 6, 7, or 8 diffraction peaks selected from 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, and 18.126±0.200° in a powder X-ray diffraction spectrum expressed in 2θ angles.

24. 23. A C-type crystal of the compound of formula (I) according to claim 22, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, and 18.126±0.200°.

25. The above-mentioned C-type crystal has a powder X-ray diffraction spectrum expressed as 2θ angles of 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, 12.988±0.200°, 14.821±0.200°, and 16.182±0.200°.

23. A C-type crystal of the compound of formula (I) according to claim 22, characterized in that it contains at least 12, 13, 14, 15, or 16 diffraction peaks selected from the group consisting of 200°, 18.126±0.200°, 18.545±0.200°, 19.474±0.200°, 20.199±0.200°, 24.294±0.200°, and 25.697±0.200°.

26. The powder X-ray diffraction spectrum thereof exhibited the following 2θ angles: 4.883±0.200°, 6.334±0.200°, 8.399±0.200°, 8.928±0.200°, 9.435±0.200°, 11.168±0.200°, 12.060±0.200°, 12.988±0.200°, 14.821±0.200°, 23. A C-type crystal of the compound of formula (I) according to claim 22, having characteristic diffraction peaks at 16.182±0.200°, 18.126±0.200°, 18.545±0.200°, 19.474±0.200°, 20.199±0.200°, 24.294±0.200°, and 25.697±0.200°.

27. 23. The C-type crystal of the compound of formula (I) according to claim 22, characterized in that the powder X-ray diffraction spectrum of the C-type crystal is as shown in Figure 7.

28. 23. The C-type crystal of the compound represented by formula (I) according to claim 22, characterized in that the differential scanning calorimetry curve of the C-type crystal has an endothermic peak value at 113.3°C ± 3°C.

29. 23. The C-type crystal of the compound of formula (I) according to claim 22, whose differential scanning calorimetry curve is as shown in Figure 8.

30. The C-type crystal of the compound of formula (I) according to claim 22, characterized in that the thermogravimetric analysis curve of the C-type crystal shows a weight loss of 3.52% at 110.0°C ± 3°C and a weight loss of 10.58% at 150.0°C ± 3°C.

31. 23. The C-type crystal of the compound represented by formula (I) according to claim 22, whose thermogravimetric analysis curve is as shown in Figure 9.

32. The C-type crystal of the compound of formula (I) according to claim 22, wherein the C-type crystal may exist in the form of a solvate crystal.

33. A D-type crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.136±0.200°, 9.908±0.200°, 17.787±0.200°, 18.329±0.200°, and 24.298±0.200°. 【Chemistry 4】

34. 34. The D-type crystal of the compound of formula (I) according to claim 33, characterized in that the powder X-ray diffraction spectrum of the D-type crystal is as shown in Figure 10.

35. 34. The D-type crystal of the compound represented by formula (I) according to claim 33, characterized in that the D-type crystal has an endothermic peak at 113.3°C ± 3°C in a differential scanning calorimetric curve.

36. 34. The D-type crystal of the compound represented by formula (I) according to claim 33, whose differential scanning calorimetry curve is as shown in Figure 11.

37. The D-type crystal of the compound represented by formula (I) according to claim 33, characterized in that the thermogravimetric analysis curve of the D-type crystal shows a weight loss of 6.59% at 90.0°C ± 3°C and a weight loss of 15.60% at 150.0°C ± 3°C.

38. 34. The D-type crystal of the compound represented by formula (I) according to claim 33, whose thermogravimetric analysis curve is as shown in Figure 12.

39. The D-type crystal of the compound of formula (I) according to claim 33, wherein the D-type crystal may exist in the form of a solvate crystal.

40. A type E crystal of the compound represented by formula (I), characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 5.987±0.200°, 9.928±0.200°, 11.998±0.200°, 13.499±0.200°, and 18.044±0.200°. 【Chemistry 5】

41. 41. The E-type crystal of the compound represented by formula (I) according to claim 40, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 5.987±0.200°, 9.928±0.200°, 11.998±0.200°, 13.499±0.200°, 18.044±0.200°, 19.492±0.200°, and 25.173±0.200°.

42. 41. The E-type crystal of the compound of formula (I) according to claim 40, whose XRPD spectrum is as shown in Figure 13.

43. 41. The E-type crystal of the compound represented by formula (I) according to claim 40, characterized in that the differential scanning calorimetry curve of the E-type crystal has an endothermic peak value at 113.7°C ± 3°C.

44. 41. The E-type crystal of the compound represented by formula (I) according to claim 40, whose DSC spectrum is as shown in Figure 14.

45. The E-type crystal of the compound of formula (I) according to claim 40, characterized in that the thermogravimetric analysis curve of the E-type crystal shows a weight loss of 13.97% at 150.0°C ± 3°C.

46. 41. The E-type crystal of the compound represented by formula (I) according to claim 40, whose thermogravimetric analysis curve is as shown in Figure 15.

47. The E-type crystal of the compound of formula (I) according to claim 40, wherein the E-type crystal may exist in the form of a solvate crystal.

48. A crystalline composition comprising the A-type crystal according to any one of claims 1 to 10, the B-type crystal according to any one of claims 11 to 21, the C-type crystal according to any one of claims 22 to 32, the D-type crystal according to any one of claims 33 to 39, or the E-type crystal according to any one of claims 40 to 47.

49. A pharmaceutical composition comprising the A-type crystal according to any one of claims 1 to 10, the B-type crystal according to any one of claims 11 to 21, the C-type crystal according to any one of claims 22 to 32, the D-type crystal according to any one of claims 33 to 39, or the E-type crystal composition according to any one of claims 40 to 47.

50. Use of the A-type crystal according to any one of claims 1 to 10, the B-type crystal according to any one of claims 11 to 21, the C-type crystal according to any one of claims 22 to 32, the D-type crystal according to any one of claims 33 to 39, the E-type crystal according to any one of claims 40 to 47, the crystalline composition according to claim 48, or the pharmaceutical composition according to claim 49 in the manufacture of a medicament for the treatment of a disease associated with coronavirus infection.

51. 51. The use according to claim 50, wherein the disease associated with coronavirus infection is SARS-CoV-2 virus infection.

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