Crystals and Uses of Viral Protease Inhibitors

Abstract

Crystallization and Use of Viral Protease Inhibitors. Specifically, this invention relates to the crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof and its use. The invention also relates to methods for preparing the crystalline form, as well as compositions containing the crystals and pharmaceutical compositions. The crystalline form is stable, has high yield, involves mild crystallization conditions, is suitable for industrial production, and can fully meet the requirements of the pharmaceutical industry.

Description

Technical Field

[0001] The present invention relates to crystals of anti-coronavirus protease inhibitor compounds and methods for their preparation, as well as compositions or pharmaceutical compositions containing the crystals and their uses. The present invention also relates to crystalline salts of protease inhibitor compounds including hydrochloride, fumarate, hippurate, malonate, methanesulfonate, benzenesulfonate and p-toluenesulfonate, methods for preparing the above salts, and compositions or pharmaceutical compositions containing the above crystalline salts and their uses.

Background Art

[0002] More than 70% of the genomic sequence of coronaviruses encodes 16 non-structural proteins (nsps) named nsp1 to nsp16. These 16 nsps are translated into two long polypeptide chains pp1a and pp1ab, from which individual nsps are generated by proteolysis. Hydrolysis is catalyzed by two proteases among 16 nsps - nsp3 and nsp5. Nsp5, also known as 3CLpro, catalyzes the hydrolysis of the peptide bond linking nsp4 to nsp16. A catalytic dyad consisting of His41 and Cys145 exists at the catalytic center of 3CLpro. The mechanism of action and protein sequence of the catalytic domain of 3CLpro are conserved among different coronaviruses. The sequence identity between 3CLpro of SARS-CoV-2 and 3CLpro of SARS-CoV reaches 96%. By inhibiting the protease activity of 3CLpro, the release of nsp4 to nsp16, which is essential for coronavirus replication, will be blocked. Therefore, 3CLpro is an important target for the development of anti-coronavirus drugs.

[0003] International Publication No. 2022150962 discloses compounds useful as inhibitors of coronavirus 3CLpro, including compound 1 described in the present invention. The contents disclosed in this patent document are incorporated in their entirety by reference. The compounds disclosed herein bind non-covalently to the catalytic pocket of 3CLpro, competitively inhibiting the binding of 3CLpro substrates, thereby reducing or inhibiting the activity of 3CLpro. Furthermore, these compounds can reduce or inhibit coronavirus replication.

[0004] The sequence associated with the catalytic pocket of coronavirus 3CLpro is highly conserved, and the recognized substrate also exhibits high conservation. Therefore, the compound may be a pan inhibitor of coronavirus 3CLpro that exerts therapeutic effects against infections caused by coronaviruses, including SARS-CoV-2, SARS-CoV, and MERS-CoV.

[0005] Those skilled in the art understand that the crystalline or salt form of a drug, and the crystalline form of a salt, significantly affect various pharmaceutical properties, including solubility, hygroscopicity, solid-state stability, and bioavailability. Therefore, there is a need for crystalline and / or salt forms of pharmaceutical compounds suitable for drug development, preparation, storage, and use. [Overview of the project]

[0006] During research and development experiments, the inventors of the present invention surprisingly found that crystalline compound 1 and certain pharmaceutically acceptable salts thereof exhibit properties particularly suitable for drug development. Therefore, the object of the present invention is to provide drug development, pharmaceutical compositions including the crystalline form, and crystalline forms of compound 1 or pharmaceutically acceptable salts thereof suitable for the medical use of the crystalline form. The name of compound 1 described in the present invention is N-((1S,2R)-2-((4-bromo-2-(methylcarbamoyl)-6-nitrophenyl)amino)cyclohexyl)isoquinoline-4-carboxamide, and its structure is as follows. [ka]

[0007] The present invention can be described in different aspects. Each of the inventions and their embodiments described in these aspects is independent of the others but is interrelated, and collectively constitutes the content of the present invention.

[0008] In a first aspect, the present invention provides a crystalline form of compound 1 or a pharmaceutically acceptable salt thereof.

[0009] In some embodiments, the present invention provides crystals of compound 1 in its free form.

[0010] In some embodiments, the present invention provides crystalline pharmaceutically acceptable salts of compound 1, where the salt is selected from hydrochloride, fumarate, hiprate, malonate, methanesulfonate, benzenesulfonate, and p-toluenesulfonate of compound 1.

[0011] In a second aspect, the present invention provides a composition comprising a crystalline form according to any embodiment of the first aspect of the present invention, wherein the crystalline form accounts for more than 50%, preferably more than 75%, more preferably more than 90%, and most preferably more than 95% of the mass of the composition.

[0012] In a third aspect, the present invention provides a pharmaceutical composition comprising a crystalline form according to any embodiment of the first aspect of the present invention and at least one pharmaceutically acceptable carrier.

[0013] In a fourth aspect, the present invention provides the use of a crystalline form according to any embodiment of the first aspect of the present invention in the preparation of a pharmaceutical product for treating a disease caused by coronavirus or its symptoms.

[0014] In a fifth aspect, the present invention provides a crystalline form according to any embodiment of the first aspect of the present invention, a composition according to the second aspect of the present invention, or a pharmaceutical composition according to the third aspect of the present invention, for use as a pharmaceutical.

[0015] In a sixth aspect, the present invention provides a crystalline form according to any embodiment of the first aspect of the present invention, a composition according to the second aspect of the present invention, or a pharmaceutical composition according to the third aspect of the present invention for use in the treatment of diseases caused by coronavirus or its symptoms.

[0016] In a seventh aspect, the present invention provides a method for treating a disease caused by coronavirus or its symptoms in a subject, comprising the step of administering a therapeutically effective amount of a crystalline form according to any embodiment of the first aspect of the present invention to the subject.

[0017] The crystalline forms of Compound 1 or its pharmaceutically acceptable salts provided by the present invention have been experimentally demonstrated to exhibit excellent performance in solubility, hygroscopicity, stability, and one or more other properties, possessing desirable pharmaceutical properties and showing very promising drug properties.

[0018] The crystalline forms of Compound 1 or its pharmaceutically acceptable salts provided by the present invention have high yields and can be prepared under mild crystallization conditions. They are suitable for industrial production and can fully meet the needs of the pharmaceutical industry. [Brief explanation of the drawing]

[0019] [Figure 1A] This figure shows the XRPD pattern of crystalline form A of compound 1 in its free form. [Figure 1B] This figure shows the TGA curve of crystalline form A of compound 1 in its free form. [Figure 1C] This figure shows the DSC curve of crystalline form A of compound 1 in its free form. [Figure 2A] This figure shows the XRPD pattern of crystalline form B of compound 1 in its free form. [Figure 2B] It is a figure showing the TGA curve of crystalline form B of compound 1 in free form. [Figure 2C] It is a figure showing the DSC curve of crystalline form B of compound 1 in free form. [Figure 3] It is a figure showing the XRPD pattern of crystalline form C of compound 1 in free form. [Figure 4A] It is a figure showing the XRPD pattern of crystalline form A of the hydrochloride salt of compound 1. [Figure 4B] It is a figure showing the TGA curve of crystalline form A of the hydrochloride salt of compound 1. [Figure 4C] It is a figure showing the DSC curve of crystalline form A of the hydrochloride salt of compound 1. [Figure 5A] It is a figure showing the XRPD pattern of crystalline form B of the hydrochloride salt of compound 1. [Figure 5B] It is a figure showing the TGA curve of crystalline form B of the hydrochloride salt of compound 1. [Figure 5C] It is a figure showing the DSC curve of crystalline form B of the hydrochloride salt of compound 1. [Figure 6A] It is a figure showing the XRPD pattern of crystalline form A of the fumarate salt of compound 1. [Figure 6B] It is a figure showing the TGA / DSC curve of crystalline form A of the fumarate salt of compound 1. [Figure 7A] It is a figure showing the XRPD pattern of crystalline form A of the hippurate salt of compound 1. [Figure 7B] It is a figure showing the TGA curve of crystalline form A of the hippurate salt of compound 1. [Figure 7C] It is a figure showing the DSC curve of crystalline form A of the hippurate salt of compound 1. [Figure 8A] It is a figure showing the XRPD pattern of crystalline form A of the malonate salt of compound 1. [Figure 8B] It is a figure showing the TGA curve of crystalline form A of the malonate salt of compound 1. [Figure 8C] It is a figure showing the DSC curve of crystalline form A of the malonate salt of compound 1. [Figure 9A]This figure shows the XRPD pattern of crystalline form A of the methanesulfonate salt of compound 1. [Figure 9B] This figure shows the TGA curve for crystalline form A of the methanesulfonate salt of compound 1. [Figure 9C] This figure shows the DSC curve of crystalline form A of the methanesulfonate salt of compound 1. [Figure 10A] This figure shows the XRPD pattern of crystalline form A of the benzenesulfonate salt of compound 1. [Figure 10B] This figure shows the TGA curve for crystalline form A of the benzenesulfonate salt of compound 1. [Figure 10C] This figure shows the DSC curve of crystalline form A of the benzenesulfonate salt of compound 1. [Figure 11A] This figure shows the XRPD pattern of crystalline form B of the benzenesulfonate salt of compound 1. [Figure 11B] This figure shows the TGA curve for crystalline form B of the benzenesulfonate salt of compound 1. [Figure 11C] This figure shows the DSC curve of crystalline form B of the benzenesulfonate salt of compound 1. [Figure 12A] This figure shows the XRPD pattern of crystalline form A of the p-toluenesulfonate salt of compound 1. [Figure 12B] This figure shows the TGA curve for crystalline form A of the p-toluenesulfonate salt of compound 1. [Figure 12C] This figure shows the DSC curve of crystalline form A of the p-toluenesulfonate salt of compound 1. [Figure 13A] This figure shows the XRPD pattern of crystalline form B of the p-toluenesulfonate salt of compound 1. [Figure 13B] This figure shows the TGA curve for crystalline form B of the p-toluenesulfonate salt of compound 1. [Figure 13C] This figure shows the DSC curve of crystalline form B of the p-toluenesulfonate salt of compound 1. [Modes for carrying out the invention]

[0020] 1.Definition Unless otherwise specified, the following terms and phrases used in this invention have the following meanings.

[0021] As used herein, the term “coronavirus” refers to a group of related RNA viruses that cause diseases in mammals and birds, typically respiratory infections in humans and birds, which may range from mild to fatal diarrhea in cattle and pigs, as well as hepatitis and encephalomyelitis in mice. Coronaviruses are enveloped viruses with a positive-sense single-stranded RNA genome and a helical nucleocapsid. They have characteristic rod-shaped spikes protruding from their surface, and in electron micrographs, they resemble the shape of the sun's corona from which the name coronavirus is derived. The category of deadly coronaviruses includes coronaviruses that cause diseases such as SARS (Severe Acute Respiratory Syndrome), MERS (Middle East Respiratory Syndrome), and COVID-19 (Severe Acute Respiratory Syndrome, an infectious disease caused by coronavirus 2).

[0022] As used herein, the terms “pharmaceutically acceptable” and “medically acceptable” refer to components that, within the bounds of sound medical judgment, are suitable for contact with human and other mammalian tissues without causing excessive toxicity, irritation, allergic reactions, etc., and that have a reasonable benefit / risk ratio. A “pharmaceutically acceptable salt” refers to any salt that is nontoxic to the recipient and can directly or indirectly provide the compound of the present invention to the recipient after administration. Suitable pharmaceutically acceptable salts are disclosed, for example, in SMBerge et al. in J. Pharmaceutical Sciences, 1977, 66, pp. 1-19. For example, pharmaceutically acceptable salts of compound 1 include, but are not limited to, sulfate, pyrosulfate, deuterated sulfate, sulfite, tetrasulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, capphosphate, caprilate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, subephosphat, sebacate, fumarate, maleate, and buty-1,4-geoate. Examples include hexyn-1,6-geoate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenyl acetate, phenyl propionate, phenyl butyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and other salts of compound 1.

[0023] As used herein, the terms “subject,” “patient,” or “recipient” refer to animals, preferably mammals, that are susceptible to coronavirus infection, and in particular humans.

[0024] The term “therapeutic dose” as used herein refers to the amount of Compound 1 or a pharmaceutically acceptable salt thereof required to produce the intended therapeutic effect (e.g., improvement of disease and / or symptoms caused by coronavirus, reduction of the severity of disease and / or symptoms caused by coronavirus, and / or delay of the progression of disease and / or symptoms caused by coronavirus). The exact amount as a therapeutic dose depends on the therapeutic purpose and can be determined by those skilled in the art using known techniques.

[0025] As used herein, the term “treatment” includes, but is not limited to, the complete or partial relief or cure of the disease caused by coronavirus, and the reduction of the risk of the disease and / or symptoms caused by coronavirus. Improvement or reduction of the severity of any of these symptoms can be evaluated according to methods and techniques known in the art.

[0026] As used herein, the term "room temperature" means ambient temperature, for example, 10 to 40°C, preferably 15 to 35°C, more preferably 20 to 30°C, for example, about 25°C.

[0027] When used herein, the terms “about” and “approximately” include, when used in conjunction with a number such as a percentage, the specified number and a numerical range recognized by those skilled in the art (e.g., ±10%; preferably ±5%; more preferably ±2%; most preferably ±1%).

[0028] As used herein, the phrases "having a characteristic diffraction peak at approximately the following 2θ angle..." and "XRPD pattern having a diffraction peak at approximately... 2θ angle" mean that the powder X-ray diffraction (XRPD) pattern has diffraction peaks within a range of ±0.5°, preferably ±0.2°, and more preferably ±0.1° of the indicated 2θ angle.

[0029] As used herein, the phrase "...°C" means a range around that temperature. For example, the endothermic signal "65.3°C" means an endothermic signal within the range of 65.3°C ± 1°C, preferably ± 0.5°C, and more preferably ± 0.2°C.

[0030] 2. The crystal of the present invention In some embodiments, the present invention provides a crystalline form A of compound 1 in its free form, which exhibits a powder X-ray diffraction (XRPD) pattern having characteristic diffraction peaks at approximately the following 2θ angles: 9.32, 10.39, 13.18, 19.76, and 23.84.

[0031] In some embodiments, the XRPD pattern of crystalline form A of compound 1 in its free form has characteristic diffraction peaks at approximately the following 2θ angles: 9.32, 10.39, 13.18, 16.11, 16.78, 19.23, 19.76, 20.30, 20.84, 23.38, and 23.84.

[0032] In some embodiments, the XRPD pattern of crystalline form A of compound 1 in its free form has characteristic diffraction peaks at approximately the following 2θ angles: 6.51, 9.32, 10.39, 11.30, 13.18, 14.67, 16.11, 16.78, 17.98, 19.23, 19.76, 20.30, 20.84, 21.21, 21.95, 23.38, 23.84, 24.64, 25.95, 27.30, 28.08, 30.23, 31.44, 33.93, 36.56, and 38.72.

[0033] In some embodiments, the XRPD pattern of crystalline form A of compound 1 in its free form is substantially as shown in Figure 1A.

[0034] In some embodiments, the thermogravimetric analysis (TGA) results of crystalline form A of compound 1 in its free form are substantially as shown in Figure 1B, with a stepwise weight loss of approximately 4.62% when the sample is heated to approximately 100°C.

[0035] In some embodiments, differential scanning calorimetry (DSC) results of crystalline form A of compound 1 in its free form are substantially as shown in Figure 1C, and have two endothermic signals at approximately 107.8°C and 129.6°C (peak temperature).

[0036] In some embodiments, the present invention provides a crystalline form B of compound 1 in its free form, exhibiting an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 6.76, 9.07, 13.50, 20.33, and 22.95.

[0037] In some embodiments, the XRPD pattern of crystalline form B of compound 1 in its free form has characteristic diffraction peaks at approximately the following 2θ angles: 6.76, 9.07, 13.50, 13.89, 15.94, 19.10, 20.16, 20.33, 21.03, 21.68, 22.36, 22.95, 24.10, and 27.14.

[0038] In some embodiments, the XRPD pattern of crystalline form B of compound 1 in its free form has characteristic diffraction peaks at approximately the following 2θ angles: 6.76, 8.40, 9.07, 10.79, 13.50, 13.89, 14.61, 15.94, 17.10, 19.10, 20.16, 20.33, 21.03, 21.31, 21.68, 22.00, 22.36, 22.95, 24.10, 24.46, 25.13, 26.53, 27.14, 27.75, 28.46, 29.39, 30.78, 31.98, 34.20, 35.70, and 38.80.

[0039] In some embodiments, the XRPD pattern of crystalline form B of compound 1 in its free form is substantially as shown in Figure 2A.

[0040] In some embodiments, the TGA results for crystalline form B of compound 1 in its free form are substantially as shown in Figure 2B, with a weight loss of approximately 3.09% when the sample is heated to approximately 150°C.

[0041] In some embodiments, the DSC characterization results of crystalline form B of compound 1 in its free form are substantially as shown in Figure 2C, exhibiting four endothermic signals at approximately 50.5°C, 87.0°C, 129.5°C, and 148.4°C (peak temperature). When crystalline form B of compound 1 in its free form is heated to approximately 128°C and then cooled to room temperature, the XRPD results indicate that the crystalline form remains unchanged before and after heating, and the sample melts after being heated to approximately 155°C.

[0042] In some embodiments, the present invention provides a crystalline form C of compound 1 in its free form, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 8.64, 9.23, and 12.99.

[0043] In some embodiments, the XRPD pattern of the crystalline form C of compound 1 in its free form has characteristic diffraction peaks at approximately the following 2θ angles: 8.64, 9.23, 12.99, and 20.29.

[0044] In some embodiments, the XRPD results for the crystalline form C of compound 1 in its free form are substantially as shown in Figure 3.

[0045] In some embodiments, the present invention provides a crystalline form A of the hydrochloride salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 7.84, 11.11, 18.71, 21.24, and 23.72.

[0046] In some embodiments, the XRPD pattern of crystalline morphology A of the hydrochloride salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 7.84, 11.11, 13.54, 18.71, 19.70, 21.24, 22.29, 23.72, and 26.03.

[0047] In some embodiments, the XRPD pattern of crystalline morphology A of the hydrochloride salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 7.84, 11.11, 12.74, 13.54, 15.07, 15.67, 18.22, 18.71, 19.70, 20.10, 21.24, 21.99, 22.29, 22.81, 23.72, 25.63, 26.03, 26.23, 26.91, 27.42, 28.22, 29.12, 29.94, 30.37, 32.50, 33.68, 34.70, 37.64, and 39.12.

[0048] In some embodiments, the XRPD results for crystalline form A of the hydrochloride salt of compound 1 are substantially as shown in Figure 4A.

[0049] In some embodiments, the TGA results for crystalline form A of the hydrochloride salt of compound 1 are substantially as shown in Figure 4B, with a weight loss of approximately 1.35% when the sample is heated to approximately 150°C.

[0050] In some embodiments, the DSC results for crystalline form A of the hydrochloride salt of compound 1 are substantially as shown in Figure 4C, and have one endothermic signal at approximately 238.6°C (peak temperature).

[0051] In some embodiments, the acid-to-base molar ratio of crystalline form A of the hydrochloride salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0052] In some embodiments, the present invention provides crystalline form B of the hydrochloride salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 12.22, 18.37, 22.99, 24.11, and 25.59.

[0053] In some embodiments, the XRPD pattern of crystalline form B of the hydrochloride salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 12.22, 13.72, 18.37, 18.79, 20.28, 21.39, 22.99, 24.11, 25.03, 25.59, 26.92, and 28.27.

[0054] In some embodiments, the XRPD pattern of crystalline form B of the hydrochloride salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 2.22, 13.72, 14.10, 15.04, 17.15, 18.37, 18.79, 19.26, 20.28, 20.93, 21.39, 21.98, 22.99, 24.11, 24.64, 25.03, 25.59, 26.03, 26.45, 26.92, 28.27, 29.83, 30.77, 31.96, 33.05, 35.43 and 37.20.

[0055] In some embodiments, the XRPD results for crystalline form B of the hydrochloride salt of compound 1 are substantially as shown in Figure 5A.

[0056] In some embodiments, the TGA results for crystalline form B of the hydrochloride salt of compound 1 are substantially as shown in Figure 5B, with a weight loss of approximately 1.03% when the sample is heated to approximately 150°C.

[0057] In some embodiments, the DSC results for crystalline form B of the hydrochloride salt of compound 1 are substantially as shown in Figure 5C, and have one endothermic signal at approximately 248.7°C (peak temperature).

[0058] In some embodiments, the acid-to-base molar ratio of crystalline form B of the hydrochloride salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0059] In some embodiments, the present invention provides a crystalline form A of the fumarate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 9.19, 10.92, 18.66, 19.48, and 20.01.

[0060] In some embodiments, the XRPD pattern of crystalline form A of the humarate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 9.19, 10.92, 11.97, 18.66, 18.90, 19.48, 20.01, 20.21, 23.28, 23.49, and 23.68.

[0061] In some embodiments, the XRPD pattern of crystalline form A of the fumarate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 5.91, 9.19, 10.00, 10.92, 11.97, 12.76, 14.60, 15.94, 17.35, 18.66, 18.90, 19.48, 20.01, 20.21, 20.49, 21.21, 21.94, 23.28, 23.49, 23.68, 24.34, 24.95, 25.70, 26.21, 27.30, 27.98, 28.43, 29.21, 29.72, 30.41, 31.21, 32.19, 33.44, 34.94 and 36.77.

[0062] In some embodiments, the XRPD results for crystalline form A of the humarate salt of compound 1 are substantially as shown in Figure 6A.

[0063] In some embodiments, the TGA / DSC results for crystalline form A of the fumarate salt of compound 1 are substantially as shown in Figure 6B, with a weight loss of about 1.10% when the sample is heated to about 160°C and a sharp endothermic peak at about 187°C (peak temperature).

[0064] In some embodiments, the acid-to-base molar ratio of crystalline form A of the humarate salt of compound 1 is 1:0.5 to 2, preferably about 1:0.9 or about 1:2.

[0065] In some embodiments, the present invention provides a crystalline form A of the hippurate of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 4.90, 9.09, 10.73, 19.56, and 21.21.

[0066] In some embodiments, the XRPD pattern of crystalline form A of the hippurate of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 4.90, 9.09, 10.73, 14.64, 19.56, 19.82, 21.01, 21.21, 22.74, 23.96, and 24.84.

[0067] In some embodiments, the XRPD pattern of crystalline morphology A of the hippurate of compound 1 is approximately the following 2θ angles: 4.90, 9.09, 10.73, 11.74, 13.06, 13.59, 13.87, 14.64, 14.99, 17.10, 17.34, 18.02, 18.41, 18.74, 19.56, 19.82, 20.08, 20.28, 21.01, 21.21 It has characteristic diffraction peaks at 21.65, 22.22, 22.74, 23.09, 23.96, 24.50, 24.84, 25.22, 25.44, 25.67, 26.17, 26.57, 27.25, 28.03, 28.81, 29.09, 29.47, 31.46, 32.46, 34.09, 35.06, 35.77, 36.18, 37.25 and 38.90.

[0068] In some embodiments, the XRPD results for crystalline form A of the hippurate of compound 1 are substantially as shown in Figure 7A.

[0069] In some embodiments, the TGA results for crystalline form A of the hippurate of compound 1 are substantially as shown in Figure 7B, with a weight loss of approximately 0.72% when the sample is heated to approximately 150°C.

[0070] In some embodiments, the DSC results for crystalline form A of the hippurate of compound 1 are substantially as shown in Figure 7C, and have one sharp endothermic peak at approximately 151.8°C (peak temperature).

[0071] In some embodiments, the acid-to-base molar ratio of the crystalline form A of the hippurate of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0072] In some embodiments, the present invention provides a crystalline form A of the malonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 5.36, 8.37, and 21.39.

[0073] In some embodiments, the XRPD pattern of crystalline form A of the malonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 5.36, 8.37, 20.49, 21.39 and 23.75.

[0074] In some embodiments, the XRPD pattern of crystalline form A of the malonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 5.36, 8.37, 9.90, 11.54, 16.99, 20.49, 21.39, 22.50, 23.75, 25.09, and 27.35.

[0075] In some embodiments, the XRPD results for crystalline form A of the malonate salt of compound 1 are substantially as shown in Figure 8A.

[0076] In some embodiments, the TGA results for crystalline form A of the malonate salt of compound 1 are substantially as shown in Figure 8B, with a weight loss of about 3.26% when the sample is heated to about 100°C and about 10.83% when further heated to about 180°C.

[0077] In some embodiments, the DSC results for crystalline form A of the malonate salt of compound 1 are substantially as shown in Figure 8C, and have three endothermic signals at approximately 53.0°C, 110.6°C, and 150.9°C (peak temperature).

[0078] In some embodiments, the acid-to-base molar ratio of crystalline form A of the malonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0079] In some embodiments, the present invention provides a crystalline form A of the methanesulfonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 8.40, 16.95, and 21.30.

[0080] In some embodiments, the XRPD pattern of crystalline form A of the methanesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.40, 11.59, 16.95, 21.30, and 25.31.

[0081] In some embodiments, the XRPD results for crystalline form A of the methanesulfonate salt of compound 1 are substantially as shown in Figure 9A.

[0082] In some embodiments, the TGA results for crystalline form A of the methanesulfonate salt of compound 1 are substantially as shown in Figure 9B, with a stepwise weight loss of about 4.43% when the sample is heated to about 160°C.

[0083] In some embodiments, the DSC results for crystalline form A of the methanesulfonate salt of compound 1 are substantially as shown in Figure 9C, and have two endothermic signals at approximately 96.7°C and 145.5°C (peak temperature).

[0084] In some embodiments, the acid-to-base molar ratio of crystalline form A of the methanesulfonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0085] In some embodiments, the present invention provides a crystalline form A of the benzenesulfonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 8.62, 19.42, 24.28, 25.62, and 26.00.

[0086] In some embodiments, the XRPD pattern of crystalline form A of the benzenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.62, 11.40, 17.24, 19.42, 20.69, 22.28, 23.62, 24.28, 25.62, 26.00 and 27.07.

[0087] In some embodiments, the XRPD results for crystalline form A of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 10A.

[0088] In some embodiments, the TGA results for crystalline form A of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 10B, with a weight loss of about 3.34% when the sample is heated to about 120°C and about 5.97% when further heated to about 180°C.

[0089] In some embodiments, the DSC results for crystalline form A of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 10C, and have two endothermic signals at approximately 65.3°C and 153.7°C (peak temperature).

[0090] In some embodiments, the acid-to-base molar ratio of crystalline form A of the benzenesulfonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0091] In some embodiments, the present invention provides crystalline form B of the benzenesulfonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 8.89, 9.93, and 22.35.

[0092] In some embodiments, the XRPD pattern of crystalline form B of the benzenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.89, 9.93, 17.75, 20.49, and 22.35.

[0093] In some embodiments, the XRPD pattern of crystalline form B of the benzenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.89, 9.93, 13.36, 15.35, 17.05, 17.75, 18.49, 20.49, 22.35, 25.58, 26.99 and 28.69.

[0094] In some embodiments, the XRPD results for crystalline form B of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 11A.

[0095] In some embodiments, the TGA results for crystalline form B of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 11B, with a weight loss of approximately 3.67% when the sample is heated to approximately 180°C.

[0096] In some embodiments, the DSC results for crystalline form B of the benzenesulfonate salt of compound 1 are substantially as shown in Figure 11C, and have one sharp endothermic peak at approximately 194.1°C (start temperature).

[0097] In some embodiments, the acid-to-base molar ratio of crystalline form B of the benzenesulfonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0098] In some embodiments, the present invention provides a crystalline form A of the p-toluenesulfonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 8.38, 19.21, 24.16, 25.75, and 27.09.

[0099] In some embodiments, the XRPD pattern of crystalline form A of the p-toluenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.38, 11.42, 16.78, 19.21, 20.55, 21.73, 22.29, 24.16, 25.75, and 27.09.

[0100] In some embodiments, the XRPD pattern of crystalline form A of the p-toluenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 8.38, 11.42, 11.93, 14.59, 16.09, 16.45, 16.78, 18.64, 19.21, 20.55, 21.73, 22.29, 24.16, 25.75, 27.09 and 30.07.

[0101] In some embodiments, the XRPD results for crystalline form A of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 12A.

[0102] In some embodiments, the TGA results for crystalline form A of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 12B, with a weight loss of about 3.64% when the sample is heated to about 130°C and about 3.17% when further heated to about 170°C.

[0103] In some embodiments, the DSC results for crystalline form A of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 12C, and have two endothermic signals at approximately 122.7°C and 150.5°C (peak temperature).

[0104] In some embodiments, the acid-to-base molar ratio of crystalline form A of the p-toluenesulfonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0105] In some embodiments, the present invention provides crystalline form B of the p-toluenesulfonate salt of compound 1, which exhibits an XRPD pattern having characteristic diffraction peaks at approximately the following 2θ angles: 7.76, 9.22, 11.99, 18.44, and 21.26.

[0106] In some embodiments, the XRPD pattern of crystalline form B of the p-toluenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 5.61, 7.76, 9.22, 11.99, 14.22, 18.44, 21.26, and 23.33.

[0107] In some embodiments, the XRPD pattern of crystalline form B of the p-toluenesulfonate salt of compound 1 has characteristic diffraction peaks at approximately the following 2θ angles: 5.61, 7.76, 9.22, 10.88, 11.99, 14.22, 15.91, 17.69, 18.44, 21.26 and 23.33.

[0108] In some embodiments, the XRPD results for crystalline form B of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 13A.

[0109] In some embodiments, the TGA results for crystalline form B of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 13B, with a weight loss of approximately 5.05% when the sample is heated to approximately 140°C.

[0110] In some embodiments, the DSC results for crystalline form B of the p-toluenesulfonate salt of compound 1 are substantially as shown in Figure 13C, and have one endothermic signal at approximately 154.6°C (peak temperature).

[0111] In some embodiments, the acid-to-base molar ratio of crystalline form A of the p-toluenesulfonate salt of compound 1 is 1:0.5 to 2, preferably 1:0.9 to 1.1, and more preferably 1:1.

[0112] 3. Preparation of the crystal of the present invention Crystal morphologies according to any embodiment of the first aspect of the present invention can be prepared in combination with common technical knowledge and prior experimental methods in the art, according to the methods disclosed in this application or obvious alternatives thereof.

[0113] In some embodiments, the crystalline form A of the free form of compound 1 is obtained by stirring compound 1 in an aqueous solution of acetonitrile at room temperature, followed by drying to obtain crystals.

[0114] Preferably, the crystalline form A of the free form of compound 1 is obtained by stirring an amorphous sample of compound 1 in ACN / H2O (1:1, v / v) at room temperature for about 2 days, and then drying it overnight under vacuum.

[0115] In some embodiments, the crystalline form B of the free form of compound 1 is obtained by stirring compound 1 in an aqueous methanol solution at room temperature, followed by drying to obtain crystals.

[0116] Preferably, the crystalline form B of the free form of compound 1 is obtained by stirring an amorphous sample of compound 1 in MeOH / H2O (1:1, v / v) at room temperature for about 2 days, and then drying it overnight under vacuum.

[0117] In some embodiments, the crystalline form C of the free form of compound 1 is obtained by evaporating the solvent from a solution of compound 1 in an aqueous methanol solution at room temperature to obtain crystals.

[0118] In some embodiments, crystalline form A of the hydrochloride salt of compound 1 is prepared by dissolving compound 1 in acetone, adding concentrated hydrochloric acid, magnetic stirring, separation by suction filtration, and drying under vacuum at room temperature to obtain crystals.

[0119] Preferably, magnetic stirring is performed at room temperature for about 2 days, followed by drying under vacuum overnight.

[0120] In some embodiments, crystalline form B of the hydrochloride salt of compound 1 is prepared by dissolving compound 1 in acetone, adding concentrated hydrochloric acid, magnetic stirring, separation by suction filtration, and drying under vacuum at room temperature to obtain crystals.

[0121] Preferably, magnetic stirring is performed at room temperature for about 4 days, followed by drying under vacuum overnight.

[0122] In some embodiments, crystalline form A of the fumarate salt of compound 1 is obtained by stirring fumaric acid and compound 1 in dimethylammonium phosphate at room temperature to precipitate the solid, and then drying it under vacuum at room temperature.

[0123] Preferably, stirring is carried out at room temperature for about 2 days, and drying under vacuum is carried out at room temperature for about 2 days.

[0124] In some embodiments, crystalline form A of the hippurate of compound 1 is prepared by adding compound 1 to hippuric acid in acetone, magnetically stirring at room temperature, separating the solid, and drying under vacuum at room temperature to obtain crystals.

[0125] Preferably, magnetic stirring is performed for about two days, followed by drying under vacuum overnight.

[0126] In some embodiments, crystalline form A of the malonate salt of compound 1 is prepared by stirring compound 1 and malonic acid in acetone at room temperature, transferring the mixture to a low temperature until completely dissolved, adding the poor solvent n-heptane to colloidize it, and then stirring in a circulating suspension at a temperature of 50-5°C to obtain crystals.

[0127] Preferably, after stirring at room temperature for 2 days, the mixture is transferred and stirred at approximately 5°C for approximately 6 days, and then at -20°C for approximately 1 day.

[0128] In some embodiments, crystalline form A of the methanesulfonate salt of compound 1 is prepared by stirring compound 1 and methanesulfonic acid in acetone at room temperature, and then drying the resulting solid under vacuum at room temperature to obtain crystals.

[0129] Preferably, stirring is carried out at room temperature for about 2 days, and drying under vacuum is carried out at room temperature for about 2 days.

[0130] In some embodiments, crystalline form A of the benzenesulfonate salt of compound 1 is prepared by suspending and stirring a solution of compound 1 and benzenesulfonic acid in ethanol at room temperature, and drying it under vacuum at room temperature to obtain crystals.

[0131] Preferably, the suspension is stirred at room temperature for about 2 days, and the drying under vacuum is carried out at room temperature for about 2 days.

[0132] In some embodiments, crystalline form B of the benzenesulfonate salt of compound 1 is prepared by suspending compound 1 and benzenesulfonic acid in acetone at room temperature and stirring to obtain crystals.

[0133] Preferably, the suspension is stirred at room temperature for about 2 days, and the drying under vacuum is carried out at room temperature for about 2 days.

[0134] In some embodiments, crystalline form A of the p-toluenesulfonate salt of compound 1 is prepared by suspending and stirring a solution of compound 1 and p-toluenesulfonic acid in ethanol at room temperature, and drying it under vacuum at room temperature to obtain crystals.

[0135] Preferably, the suspension is stirred at room temperature for about 2 days, and then dried under vacuum for about 2 days.

[0136] In some embodiments, crystalline form B of the p-toluenesulfonate salt of compound 1 is prepared by suspending compound 1 and p-toluenesulfonic acid in ethyl acetate at room temperature, stirring, and drying under vacuum at room temperature to obtain crystals.

[0137] Preferably, the suspension is stirred at room temperature for about 2 days, and then dried under vacuum for about 2 days.

[0138] 4. Use of the crystal form of the present invention Compound 1 described in the present invention can reduce or inhibit the activity of 3CLpro and reduce or inhibit coronavirus replication. Therefore, the crystalline form of the present invention can be used as a broad-spectrum inhibitor of coronavirus 3CLpro for the treatment of infections caused by coronaviruses, including SARS-CoV-2, SARS-CoV, MERS-CoV, etc.

[0139] For medical and pharmaceutical applications, the crystalline form of the present invention can be administered to patients in the form of any standard pharmaceutical composition requiring it. The pharmaceutical compositions of the present invention include a pharmaceutically acceptable carrier which can be selected from adjuvants and vehicles. The pharmaceutically acceptable carrier used in the present invention can be selected from any suitable solvent, diluent, vehicle, dispersant, suspending agent, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder, and lubricant. Those skilled in the art can select a carrier suitable for the present invention from known pharmaceutically acceptable carriers according to the desired specific dosage form. For example, Remington: The Science and Practice of Pharmacy, 21 stThe 2005 edition (edited by DB Troy, Lippincott Williams & Wilkins, Philadelphia) and the Encyclopedia of Pharmaceutical Technology (edited by J. Swarbrick and JCBoylan, 1988-1999, and Marcel Dekker, New York) disclose various carriers for formulating pharmaceutical compositions and known techniques for preparing them. Non-limiting examples of suitable pharmaceutically acceptable carriers include ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffers (e.g., phosphates, glycine, sorbic acid, and potassium sorbate), mixtures of partial glycerides of saturated vegetable fatty acids, water, salts and electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, polyacrylate, waxes, polyethylene polyoxypropylene block polymers, lanolin, sugars (lactose, glucose, and sucrose, etc.), starches (e.g., corn starch and potato starch), cellulose and its derivatives (carbococcus). Examples of excipients include sodium methylcellulose, ethylcellulose, and cellulose acetate, tragacanth powder, malt, gelatin, talc, excipients (e.g., cocoa butter and suppository wax), oils (e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (e.g., propylene glycol and polyethylene glycol), esters (e.g., ethyl oleate and ethyl laurate), agar, buffers (e.g., magnesium hydroxide and aluminum hydroxide), alginic acid, water free of pyrogens, isotonic saline, Ringer's solution, ethanol, phosphate buffer, non-toxic compatible lubricants (e.g., sodium lauryl sulfate and magnesium stearate), colorants, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, and antioxidants.

[0140] The pharmaceutical compositions of the present invention can be administered to patients in need by any convenient route. For example, the crystalline form of compound 1 described in the present invention, its pharmaceutically acceptable salt, or the crystalline form of the salt can be formulated into a corresponding pharmaceutical preparation, which can then be administered, for example, orally, parenterally, sublingually, topically, rectally, nasally, buccally, vaginally, transdermally, by patch, pump, and other routes, or via an implanted reservoir.

[0141] For oral administration, any suitable pharmaceutical carrier commonly used in the preparation of solid formulations can be used, and the crystalline form of the present invention can be formulated into solid formulations such as syrups, suspensions, emulsions, tablets, capsules, and lozenges using formulation techniques known in the art. Examples of these carriers include magnesium stearate, starch, lactose, sucrose, and cellulose. Alternatively, any suitable pharmaceutical carrier commonly used in the preparation of liquid formulations can be used, and the crystalline form of the present invention can be formulated into liquid formulations using formulation techniques known in the art. The liquid formulation of the present invention may contain one or more suspending agents, preservatives, flavoring agents, or coloring agents.

[0142] For parenteral administration, any suitable pharmaceutical carrier commonly used in the preparation of formulations for parenteral administration can be used, and the crystalline form of the present invention can be formulated using formulation techniques known in the art into formulations suitable for parenteral administration, such as sterile injections, lyophilized powders, transdermal patches, aerosols, oral and nasal inhalants, and suppositories. Routes of parenteral administration include intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, transnasal, intrapulmonary, intrathecal, rectal, and local routes. Parenteral administration may be a continuous infusion over a given period of time. [Examples]

[0143] To better understand the scope of the present invention, further examples are provided below, along with specific examples. However, these specific examples are not intended to limit the scope of the present invention.

[0144] All solvents used in the following examples are commercially available and used without further treatment unless otherwise specified.

[0145] The following abbreviations are used in the following examples. v / v represents the volume ratio. ACN stands for acetonitrile. Acetone refers to acetone. MeOH represents methanol.  represents ethyl acetate, XRPD stands for powder X-ray diffraction. TGA stands for thermogravimetric analysis. DSC stands for Differential Scanning Calorimetry. DVS stands for Dynamic Vapor Deposition. UPLC stands for ultrafast liquid chromatography. SGF represents simulated gastric juice. FaSSIF represents fasting-simulated intestinal fluid. FeSSIF represents simulated intestinal fluid.

[0146] Example 1: Preparation of Compound 1 The amorphous form of compound 1 in its free form was prepared as a yellow solid according to the method disclosed in paragraphs

[0121] to

[0128] of International Publication No. 2022150962 and used in the following example. The specific synthesis steps are shown in steps a to d below: [ka]

[0147] The reagents and conditions for steps a to d described below are as follows: (a) 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), N,N-diisopropylethylamine (DIPEA), CH2Cl2 or dichloromethane (DCM), 0°C, 2 hours; (b) DIPEA, dimethylformamide (DMF), 80°C, 16 hours; (c) 3M hydrogen chloride-ethyl acetate (HCl·EA), CH2Cl2, 1 hour; (d) HATU, DIPEA, DMF, room temperature, 12 hours.

[0148] Step a: Synthesis of N-methyl-5-bromo-2-fluoro-3-nitrobenzamide (I-1) A solution of 5-bromo-2-fluoro-3-nitrobenzoic acid (0.8 g, 3.80 mmol) in dichloromethane (20 mL) was stirred at 0°C. Then, HATU (2.0 g, 5.25 mmol), DIPEA (1.88 mL, 11.4 mmol), and methylamine hydrochloride (0.31 g, 4.5 mmol) were added to the reaction mixture. The mixture was stirred at 0°C for 2 hours until clear. The mixture was extracted three times with dichloromethane, and the combined organic layers were washed with saturated brine. The organic phase was then dried over anhydrous Na₂SO₄ and concentrated under vacuum. Finally, the mixture was purified by chromatography to obtain compound I-1 as a yellow solid (0.8 g, yield 76%).

[0149] Step b: Synthesis of tert-butyl(2-((4-bromo-2-(methylcarbamoyl)-6-nitrophenyl)amino)cyclohexyl)carbamate (I-2) A solution of compound I-1 (0.8 g, 2.9 mmol) in DMF (15 mL) was stirred at room temperature. Then, tert-butyl((1S,2R)-2-aminocyclohexyl) carbamate (0.75 g, 3.5 mmol) (the stereoisomer of compound I-2 can be synthesized using the corresponding stereoisomer of this reagent) and DIPEA (1.44 mL, 8.7 mmol) were added to the reaction. The mixture was heated to 80°C and stirred for 16 hours. The mixture was extracted three times with ethyl acetate, and the combined organic layers were washed with saturated brine. The organic phase was then dried over anhydrous Na2SO4 and concentrated under vacuum to obtain compound I-2 as a yellow solid, which was used without further purification. [ka]

[0150] Step c: Synthesis of 2-(2-aminocyclohexyl)amino)-5-bromo-N-methyl-3-nitrobenzamide hydrochloride (I-3) A solution of compound I-2 (90 mg, 0.19 mmol) (or its corresponding stereoisomer) in anhydrous dichloromethane (6 mL) was stirred at room temperature. Then, HCl (4 mL, 3 M in ethyl acetate) was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum to obtain compound I-3 as a yellow solid, which was used without further purification.

[0151] Step d: Synthesis of N-((1S,2R)-2-((4-bromo-2-(methylcarbamoyl)-6-nitrophenyl)amino)cyclohexyl)isoquinoline-4-carboxamide The corresponding solutions of isoquinoline-4-carboxylic acid (1 equivalent) and HATU (1.5 equivalents) in anhydrous DMF (6 mL) were stirred at room temperature. Compound I-3 and DIPEA (5.0 equivalents) were then added. The mixture was stirred overnight at room temperature. The mixture was extracted three times with ethyl acetate, and the combined organic layers were washed with saturated brine. The organic phase was then dried over anhydrous Na2SO4 and concentrated under vacuum. Finally, the mixture was purified by chromatography to obtain free amorphous compound 1 as a yellow solid.

[0152] Example 2: Preparation and characterization of crystalline form A of compound 1 in its free form. The preparation steps for crystalline form A of compound 1 in its free form are as follows.

[0153] (1) 399.9 mg of free amorphous compound 1 was weighed into a 20 mL glass vial, and 10 mL of ACN / H2O (1:1) was added.

[0154] (2) The mixture was magnetically stirred at room temperature for 2 days, and then the solid was separated by centrifugation (10,000 rpm, 2 minutes).

[0155] (3) The solid was dried under vacuum at room temperature overnight.

[0156] Table 1 shows the diffraction peak data for crystalline form A of compound 1 in its free form.

[0157] Figure 1A shows the XRPD pattern of crystalline form A of compound 1 in its free form.

[0158] Figure 1B shows the TGA curve for crystalline form A of compound 1 in its free form, which exhibits a stepwise weight loss of approximately 4.62% when the sample is heated to approximately 100°C.

[0159] Figure 1C shows the DSC curve for crystalline form A of compound 1 in its free form, along with two endothermic signals at approximately 107.8°C and 129.6°C (peak temperature). [Table 1]

[0160] Example 3: Preparation and characterization of crystalline form B of compound 1 in its free form. The preparation steps for crystalline form B of compound 1 in its free form are as follows.

[0161] (1) 399.9 mg of free amorphous compound 1 was weighed into a 20 mL glass vial, and 10 mL of MeOH / H2O (1:1) was added.

[0162] (2) The mixture was magnetically stirred at room temperature for 2 days, and then the solid was separated by centrifugation (10,000 rpm, 2 minutes).

[0163] (3) The solid was dried under vacuum at room temperature overnight.

[0164] Table 2 shows the diffraction peak data for crystalline form B of compound 1 in its free form.

[0165] Figure 2A shows the XRPD pattern of crystalline form B of compound 1 in its free form.

[0166] Figure 2B shows the TGA curve for crystalline form B of compound 1 in its free form, and the weight loss when the sample is heated to approximately 150°C is approximately 3.09%.

[0167] Figure 2C shows the DSC curve of crystalline form B of compound 1 in its free form, which has four endothermic signals at approximately 50.5°C, 87.0°C, 129.5°C, and 148.4°C (peak temperature). When crystalline form B of compound 1 in its free form is heated to approximately 128°C and then cooled to room temperature, the XRPD results show that the crystalline form remains unchanged before and after heating, and that the sample melts when heated to approximately 155°C. [Table 2-1] [Table 2-2]

[0168] Example 4: Preparation and characterization of crystalline form C of compound 1 in its free form. The preparation steps for the crystalline form C of compound 1 in its free form are as follows.

[0169] Crystalline form C of free compound 1 was obtained by slowly evaporating a solution of an amorphous sample of compound 1 in methanol at room temperature. After standing for 4 days under ambient conditions, a mixed crystal was obtained consisting of crystalline form B of free compound 1 and a small amount of crystalline form C of free compound 1.

[0170] Table 3 shows the diffraction peak data for crystalline form C of compound 1 in its free form.

[0171] Figure 3 shows the XRPD pattern of crystalline form C of compound 1 in its free form. [Table 3]

[0172] Example 5: Preparation and characterization of crystalline form A of the hydrochloride salt of compound 1. The preparation steps for crystalline form A of the hydrochloride salt of compound 1 are as follows.

[0173] (1) 400.3 mg of free amorphous compound 1 was weighed into a 20 mL glass vial, and after completely dissolving it in 10 mL of acetone, 65 μL of concentrated hydrochloric acid (38%) was added.

[0174] (2) The mixture was magnetically stirred at room temperature for two days, and the solid was separated by suction filtration.

[0175] (3) The solid was dried under vacuum at room temperature overnight, and a total of 187 mg of sample (yield: 43.7%) was recovered.

[0176] Table 4 shows the diffraction peak data for crystalline form A of the hydrochloride salt of compound 1.

[0177] Figure 4A shows the XRPD pattern of crystalline form A of the hydrochloride salt of compound 1.

[0178] Figure 4B shows the TGA curve for crystalline form A of the hydrochloride salt of compound 1, and the weight loss when the sample is heated to approximately 150°C is approximately 1.35%.

[0179] The DSC curve for crystalline form A of the hydrochloride salt of compound 1 is shown in Figure 4C, along with a single endothermic signal at approximately 238.6°C (peak temperature). [Table 4]

[0180] Example 6: Preparation and characterization of crystalline form B of the hydrochloride salt of compound 1. The preparation steps for crystalline form B of the hydrochloride salt of compound 1 are as follows.

[0181] (1) 3.0 g of free amorphous compound 1 was weighed into a 50 mL glass vial, 40 mL of acetone was added to completely dissolve it, and then 512 μL of concentrated hydrochloric acid was added.

[0182] (2) The mixture was magnetically stirred at room temperature for 4 days, and the solid was separated by suction filtration.

[0183] (3) The solid was dried under vacuum at room temperature overnight, and a total of 1.72 g of sample (yield: 57.3%) was recovered.

[0184] Table 5 shows the diffraction peak data for crystalline form B of the hydrochloride salt of compound 1.

[0185] Figure 5A shows the XRPD pattern of crystalline form B of the hydrochloride salt of compound 1.

[0186] Figure 5B shows the TGA curve for crystalline form B of the hydrochloride salt of compound 1, and the weight loss when the sample is heated to approximately 150°C is approximately 1.03%.

[0187] The DSC curve for crystalline form B of the hydrochloride salt of compound 1 is shown in Figure 5C, along with a single endothermic signal at approximately 248.7°C (peak temperature). [Table 5]

[0188] Example 7: Preparation and characterization of crystalline form A of the fumarate salt of compound 1. The preparation steps for crystalline form A of the fumarate salt of compound 1 are as follows.

[0189] (1) Approximately 160 mg of free amorphous compound 1 and 18.5 mg of fumaric acid were weighed into a 3 mL glass vial, and 2 mL of  was added.

[0190] (2) The mixture was magnetically stirred at room temperature for 3 hours, after which seed crystals were added.

[0191] (3) The mixture was magnetically stirred at room temperature for a further 4 hours, and then the solid was separated by centrifugation (10,000 rpm, 2 minutes).

[0192] (4) The solid was dried under vacuum at room temperature overnight, and a total of 140 mg of sample (yield: 78.4%) was recovered.

[0193] or, (1) 399.5 mg of free amorphous compound 1 and 88.0 mg of fumaric acid were weighed into a 20 mL glass vial, and 10 mL of  was added.

[0194] (2) The mixture was magnetically stirred at room temperature for two days, and the solid was separated by suction filtration.

[0195] (3) The solid was dried under vacuum at room temperature overnight, and a total of 292 mg of sample (yield: 59.9%) was recovered.

[0196] Table 6 shows the diffraction peak data for crystalline form A of the fumarate salt of compound 1.

[0197] Figure 6A shows the XRPD pattern of crystalline form A of the humarate salt of compound 1.

[0198] Figure 6B shows the TGA / DSC curve for crystalline form A of the fumarate salt of compound 1. When the sample is heated to approximately 160°C, there is a weight loss of approximately 1.10% and a sharp endothermic peak at approximately 187°C (peak temperature). [Table 6]

[0199] Example 8: Preparation and characterization of crystalline form A of hippurate of compound 1. The preparation steps for crystalline form A of the hippurate of compound 1 are as follows.

[0200] (1) 400.0 mg of free amorphous compound and 136.2 mg of hippuric acid were weighed into a 20 mL glass vial, and 10 mL of acetone was added.

[0201] (2) The mixture was magnetically stirred at room temperature for two days, and then the solid was separated by suction filtration.

[0202] (3) The solid was dried under vacuum at room temperature overnight, and a total of 303 mg of sample (yield: 61.5%) was recovered.

[0203] Table 7 shows the diffraction peak data for crystalline form A of the hippurate of compound 1.

[0204] Figure 7A shows the XRPD pattern of crystalline form A of the hippurate of compound 1.

[0205] Figure 7B shows the TGA curve for crystalline form A of the hippurate of compound 1, and the weight loss when the sample is heated to approximately 150°C is approximately 0.72%.

[0206] Figure 7C shows the DSC curve of crystalline form A of the hippurate of compound 1, which has a single sharp endothermic peak at approximately 151.8°C (peak temperature). [Table 7-1] [Table 7-2]

[0207] Example 9: Preparation and characterization of crystalline form A of the malonate salt of compound 1. The preparation steps for crystalline form A of the malonate salt of compound 1 are as follows.

[0208] (1) Approximately 400 mg of free amorphous compound and an equimolar amount of malonic acid were stirred in acetone at room temperature for 2 days.

[0209] (2) The mixture was transferred to 5°C and stirred for 6 days, then transferred to -20°C and stirred for 1 day, and the mixture remained clear.

[0210] (3) A poor solvent n-heptane was added to form a colloidal substance.

[0211] (4) The mixture was transferred to a temperature of 50-5°C and stirred in a circulating suspension to precipitate the solid.

[0212] Table 8 shows the diffraction peak data for crystalline form A of the malonate salt of compound 1.

[0213] Figure 8A shows the XRPD pattern of crystalline form A of the malonate salt of compound 1.

[0214] Figure 8B shows the TGA curve for crystalline form A of the malonate salt of compound 1. When the sample is heated to approximately 100°C, it exhibits a weight loss of approximately 3.26%, and when further heated to approximately 180°C, it exhibits a weight loss of approximately 10.83%.

[0215] The DSC curve for crystalline form A of the malonate salt of compound 1 is shown in Figure 8C, along with three endothermic signals at approximately 53.0°C, 110.6°C, and 150.9°C (peak temperature). [Table 8]

[0216] Example 10: Preparation and characterization of crystalline form A of the methanesulfonate salt of compound 1. The preparation steps for crystalline form A of the methanesulfonate salt of compound 1 are as follows.

[0217] (1) Approximately 400 mg of free amorphous compound and an equimolar amount of methanesulfonic acid in acetone were stirred at room temperature for 2 days.

[0218] (2) The obtained solid was vacuum-dried at room temperature for 2 days to obtain crystalline form A of the methanesulfonate salt.

[0219] Table 9 shows the diffraction peak data for crystalline form A of the methanesulfonate salt of compound 1.

[0220] Figure 9A shows the XRPD pattern of crystalline form A of the methanesulfonate salt of compound 1.

[0221] Figure 9B shows the TGA curve for crystalline form A of the methanesulfonate salt of compound 1, which exhibits a stepwise weight loss of approximately 4.43% when the sample is heated to approximately 160°C.

[0222] The DSC curve for crystalline form A of the methanesulfonate salt of compound 1 is shown in Figure 9C, along with two endothermic signals at approximately 96.7°C and 145.5°C (peak temperature). [Table 9]

[0223] Example 11: Preparation and characterization of crystalline form A of the benzenesulfonate salt of compound 1. The preparation steps for crystalline form A of the benzenesulfonate salt of compound 1 are as follows.

[0224] (1) Approximately 400 mg of free amorphous compound 1 and equimolar amounts of benzenesulfonic acid in EtOH / H2O (19:1, v / v) were suspended and stirred at room temperature for 2 days.

[0225] (2) The solid was dried under vacuum at room temperature for two days to obtain crystalline form A of the benzenesulfonate salt.

[0226] Table 10 shows the diffraction peak data for crystalline form A of the benzenesulfonate salt of compound 1.

[0227] Figure 10A shows the XRPD pattern of crystalline form A of the benzenesulfonate salt of compound 1.

[0228] The TGA curve of crystalline form A of the benzenesulfonate salt of Compound 1 is shown in Figure 10B, showing a weight loss of approximately 3.34% when the sample is heated to approximately 120 °C and approximately 5.97% when further heated to approximately 180 °C.

[0229] The DSC curve of crystalline form A of the benzenesulfonate salt of Compound 1 is shown in Figure 10C, showing two endothermic signals at approximately 65.3 °C and 153.7 °C (peak temperature). [Table 10]

[0230] Example 12: Preparation and Characterization of Crystalline Form B of the Benzenesulfonate Salt of Compound 1 The preparation process of crystalline form B of the benzenesulfonate salt of Compound 1 is as follows.

[0231] (1) An acetone solution of approximately 400 mg of free amorphous Compound 1 and an equimolar amount of benzenesulfonic acid was stirred in a suspended state at room temperature for 2 days.

[0232] (2) The solid was dried under vacuum at room temperature for 2 days to obtain crystalline form B of the benzenesulfonate salt.

[0233] The diffraction peak data of crystalline form B of the benzenesulfonate salt of Compound 1 are shown in Table 11.

[0234] The XRPD pattern of crystalline form B of the benzenesulfonate salt of Compound 1 is shown in Figure 11A.

[0235] The TGA curve of crystalline form B of the benzenesulfonate salt of Compound 1 is shown in Figure 11B, and the weight loss when the sample is heated to approximately 180 °C is approximately 3.67%.

[0236] The DSC curve of crystalline form B of the benzenesulfonate salt of Compound 1 is shown in Figure 11C, having one sharp endothermic peak at approximately 194.1 °C (starting temperature). [Table 11]

[0237] Example 13: Preparation and Characterization of Crystal Form A of the p-Toluenesulfonate Salt of Compound 1 The preparation of crystal form A of the p-toluenesulfonate salt of Compound 1 is as follows.

[0238] (1) Approximately 400 mg of the free amorphous Compound 1 and an equimolar amount of p-toluenesulfonic acid in EtOH / H2O (19:1, v / v) were stirred in suspension at room temperature for 2 days.

[0239] (2) The solid was dried under vacuum at room temperature for 2 days.

[0240] The diffraction peak data of crystal form A of the p-toluenesulfonate salt of Compound 1 are shown in Table 12.

[0241] The XRPD pattern of crystal form A of the p-toluenesulfonate salt of Compound 1 is shown in Figure 12A.

[0242] The TGA curve of crystal form A of the p-toluenesulfonate salt of Compound 1 is shown in Figure 12B, showing a weight loss of approximately 3.64% when the sample is heated to about 130 °C and approximately 3.17% when further heated to about 170 °C.

[0243] The DSC curve of crystal form A of the p-toluenesulfonate salt of Compound 1 is shown in Figure 12C with two endothermic signals at approximately 122.7 °C and 150.5 °C (peak temperatures). [Table 12]

[0244] Example 14: Preparation and Characterization of Crystal Form B of the p-Toluenesulfonate Salt of Compound 1 The preparation process of crystal form B of the p-toluenesulfonate salt of Compound 1 is as follows.

[0245] (1) Approximately 400 mg of the free amorphous Compound 1 and an equimolar amount of p-toluenesulfonic acid in EtOAc were stirred in suspension at room temperature for 2 days.

[0246] (2) The resulting solid was dried under vacuum at room temperature for 2 days.

[0247] The diffraction peak data of the crystalline Form B of the p-toluenesulfonate salt of Compound 1 are shown in Table 13.

[0248] The XRPD pattern of the crystalline Form B of the p-toluenesulfonate salt of Compound 1 is shown in Figure 13A.

[0249] The TGA curve of the crystalline Form B of the p-toluenesulfonate salt of Compound 1 is shown in Figure 13B, and the weight loss when the sample is heated to about 140 °C is about 5.05%.

[0250] The DSC curve of the crystalline Form B of the p-toluenesulfonate salt of Compound 1 is shown in Figure 13C together with one endothermic signal at about 154.6 °C (peak temperature).

Table 13

[0251] Example 15: Dynamic Solubility Test The solubilities of the crystalline forms obtained from Examples 2 - 14 in water (H2O) and biological vehicles (simulated gastric fluid (SGF), fasted-state simulated intestinal fluid (FaSSIF), and fed-state simulated intestinal fluid (FeSSIF)) were tested at 1 / 2 / 4 / 24 hours. The experimental results are shown in Table 14.

[0252] The specific procedure of the test is as follows.

[0253] 1) Approximately 20 - 40 mg of the crystalline forms from Examples 2 - 14 were weighed into 5 mL glass vials, and 4 mL of the corresponding media (H2O, SGF, FaSSIF, and FeSSIF) were added to each vial.

[0254] 2) The vials were mixed by rotating them on a rotary incubator at 37°C at a speed of 25 rpm. Sampling was performed at 1, 2, 4, and 24 hours.

[0255] 3) At each sampling point, approximately 0.8 mL of the suspension was placed in a centrifuge tube and separated by centrifugation (12,000 rpm, 5 minutes, 37°C).

[0256] 4) The supernatant was filtered through a PTFE membrane (pore size: 0.45 μm), the filtrate was used for solubility and pH testing, and the solid was subjected to XRPD analysis.

[0257] Experimental results: [Table 14]

[0258] The experimental results are as follows:

[0259] 1) In water (H2O), the solubility of the four salt forms is slightly higher than that of the three free forms.

[0260] 2) In SGF, the solubility of crystalline forms A and B of the hydrochloride salts is significantly higher than that of the humarate salt, hippurate salt, and the three free form samples.

[0261] 3) In FaSSIF, the solubility of crystalline form A of the humarate salt is slightly higher than that of crystalline form A / B of the hydrochloride salt, crystalline form A of the hippurate salt, crystalline form A / B of the free form, and the free form of the amorphous salt.

[0262] 4) In FeSSIF, there is no significant difference in the solubility of the free amorphous form, the free crystalline form B, the hydrochloride crystalline form A / B, the humarate salt crystalline form A, and the hippurate crystalline form A.

[0263] 5) XRPD results show that the free amorphous form undergoes morphological changes after being suspended in FeSSIF for 2 hours. Free crystalline forms A / B and the four salt forms do not undergo morphological changes in any of the media.

[0264] Example 16: Moisture absorption test In accordance with the Chinese Pharmacopoeia Guidelines for Hygroscopicity Testing of Drugs (2020 Edition), DVS tests were performed on the crystalline forms of Examples 2-14 to determine their hygroscopicity. The results of the hygroscopicity tests are shown in Table 15.

[0265] Experimental results: [Table 15]

[0266] The DVS results indicate that all tested crystalline forms exhibited little to no hygroscopicity, representing a significant advantage over the free amorphous form. Furthermore, no changes were observed in any of the crystalline forms after the DVS test.

[0267] Example 17: Solid State Stability Test Appropriate amounts of the crystalline samples from Examples 2-14 were weighed and left open for 7-10 days under conditions of 25°C / 60%RH and 40°C / 75%RH for stability testing. For the solid samples separated under different conditions, the crystalline morphology was tested using XRPD to evaluate physical stability, and the purity was tested using UPLC to evaluate chemical stability. The results of the solid state stability tests are shown in Table 16.

[0268] The experimental results are shown in the table below. [Table 16]

[0269] The solid-state stability results show that all tested crystalline samples did not show any morphological changes or purity degradation after being left open for 7-10 days at 25°C / 60%RH and 40°C / 75%RH, indicating that they all possess good physical and chemical stability under the evaluation conditions.

[0270] Equipment and methods 1. Powder X-ray diffraction (also known as powder X-ray diffractometer or XRPD) XRPD results are collected using X'Pert3 and Empyrean powder X-ray diffractometers with the scanning parameters shown in Table 17. [Table 17]

[0271] 2. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) TGA results were collected using a TA Discovery 5500 thermogravimetric analyzer, and DSC results were collected using a TA Discovery 2500 differential scanning calorimeter. The test parameters are shown in Table 18. [Table 18]

[0272] 3. Dynamic water vapor adsorption (DVS) Dynamic water vapor adsorption (DVS) curves were collected using a Surface Measurement Systems (SMS) DVS Intrinsic instrument. At 25°C, relative humidity was calibrated using the deliquescence points of LiCl, Mg(NO3)2, and KCl. The DVS test parameters are shown in Table 19. [Table 19]

[0273] 4. Ultra-high-speed liquid chromatography (UPLC) In the experiment, purity, solubility, and molar ratio were tested using a Waters H-Class UPLC ultra-high-performance liquid chromatograph under the analytical conditions shown in Tables 20 and 21. [Table 20] [Table 21]

Claims

1. The crystalline form of the compound of the following formula or a pharmaceutically acceptable salt thereof. 【Chemistry 1】

2. The crystalline form according to claim 1, wherein the salt is a salt formed by the compound with an acid selected from hydrochloric acid, fumaric acid, hippuric acid, malonic acid, methanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid.

3. The crystalline form according to claim 2, characterized in that the salt is formed by the compound and the acid in a molar ratio of 1:0.5 to 2.

4. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 9.32, 10.39, 13.18, 19.76, and 23.

84.

5. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 6.76, 9.07, 13.50, 20.33, and 22.

95.

6. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 8.64, 9.23, and 12.

99.

7. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 7.84, 11.11, 18.71, 21.24, and 23.

72.

8. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 12.22, 18.37, 22.99, 24.11, and 25.

59.

9. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 9.19, 10.92, 18.66, 19.48, and 20.

01.

10. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 4.90, 9.09, 10.73, 19.56, and 21.

21.

11. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 5.36, 8.37, and 21.

39.

12. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 8.40, 16.95, and 21.

30.

13. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 8.62, 19.42, 24.28, 25.62, and 26.

00.

14. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 8.89, 9.93, and 22.

35.

15. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 8.38, 19.21, 24.16, 25.75, and 27.

09.

16. The crystal morphology according to claim 1, characterized in that the crystal morphology exhibits an XRPD pattern having diffraction peaks at 2θ angles of approximately 7.76, 9.22, 11.99, 18.44, and 21.

26.

17. A composition comprising the crystalline form according to any one of claims 1 to 16, wherein the crystalline form accounts for more than 50%, preferably more than 75%, more preferably more than 90%, and most preferably more than 95% of the mass of the composition.

18. A pharmaceutical composition comprising a crystalline form according to any one of claims 1 to 16 and at least one pharmaceutically acceptable carrier.

19. Use of a crystalline form according to any one of claims 1 to 16 in the preparation of a pharmaceutical product for treating a disease caused by coronavirus or symptoms thereof, preferably wherein the disease is selected from COVID-19, SARS, and MERS.

20. A crystalline form according to any one of claims 1 to 16 or a composition according to claim 16, for use as a pharmaceutical product.

21. A pharmaceutical composition according to claim 18 for use in the treatment of a disease caused by coronavirus or symptoms thereof, preferably wherein the coronavirus is selected from SARS-CoV-2, SARS-CoV and MERS-CoV.

22. A method for treating a disease caused by coronavirus or its symptoms in a subject, comprising administering to the subject a therapeutically effective amount of the crystalline form described in any one of claims 1 to 16, preferably the disease being selected from COVID-19, SARS, and MERS.

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