Crystalline polymorphic substance of coronavirus 3CL protease inhibitor as well as preparation method and application of crystalline polymorphic substance

Abstract

The invention discloses a crystalline polymorphic substance of PLC-01 as well as a preparation method and application of the crystalline polymorphic substance, and belongs to the field of chemical medicines. The crystalline polymorphic substance mainly comprises a Pattern A crystal form and a Pattern B crystal form, wherein an X-ray powder diffraction pattern of the Pattern A crystal form has characteristic peaks at a series of specific positions of which the 2theta value is 7.8 + / -0.2 degrees and the like; the Pattern B crystal form has characteristic peaks at corresponding positions such as 9.0 + / -0.2 degrees and the like. Wherein the PatternA crystal form has significant advantages, has good chemical stability and physical stability, and is convenient for long-term storage of drugs; and the preparation process is simple. Due to the characteristics, the PatternA crystal form has important value in optimization and development of the medicine in the future, a new choice and a new direction are provided for research, development and production of related medicines, and development of the chemical medicine field is expected to be promoted.

Description

Technical Field

[0001] This invention belongs to the field of chemical and pharmaceutical technology, specifically relating to crystalline polymorphs of coronavirus 3CL protease inhibitors, their preparation methods, and applications. Background Technology

[0002] The coronavirus 3CL protease (3CLpro, also known as the major protease Mpro) is a key enzyme in viral replication, possessing cleavage site specificity similar to that of the microRNA virus 3C protease, and participates in the replication and transcription of progeny viruses. Composed of 306 amino acids, 3CLpro specifically recognizes and cleaves 11 cleavage sites of non-structural proteins NSP4-NSP16, releasing other non-structural proteins that play important roles in viral genome replication, transcription, and post-translational modifications. Inhibiting 3CLpro can effectively block viral RNA replication and transcription, preventing viral proliferation; therefore, 3CLpro is an ideal target for coronavirus targeted drug development.

[0003] The coronavirus 3CL protease inhibitor (PLC-01) is a 3-triazolylmethyl-1,3,5-triazine-2,4-dione compound that has been found to have a strong inhibitory effect on the coronavirus 3CL protease (3CLpro), effectively blocking viral replication and transcription. Therefore, PLC-01 shows great promise as a potential anti-coronavirus drug. However, different crystal forms of the compound often exhibit different physicochemical properties, including solubility, stability, and dissolution rate, which directly affect its efficacy and formulation quality. To fully realize the pharmacological activity and clinical therapeutic potential of PLC-01, research on its crystal forms is particularly important. Different crystal forms of PLC-01 show significant differences in physicochemical stability, duration of efficacy, and dosage control. However, the physicochemical properties of the crystal forms and the stability, hygroscopicity, and efficacy of the formulations of PLC-01 in the current technology need further optimization, which limits its clinical application value as an anti-coronavirus drug. Therefore, it is urgent to study the crystal form of PLC-01 and propose new crystal forms in order to optimize its drug performance and enhance its clinical application value as an anti-coronavirus drug. Summary of the Invention

[0004] In view of the problems in the existing technology that the physicochemical properties of the crystalline form of coronavirus 3CL protease inhibitors and the stability, hygroscopicity and efficacy of the formulation need to be optimized, as well as the problems that the preparation process is complicated and the conditions are harsh, the purpose of this invention is to provide crystalline polymorphs of coronavirus 3CL protease inhibitors, their preparation methods and applications.

[0005] To achieve the above objectives, the present invention employs the following technical solution:

[0006] In a first aspect, the present invention discloses a crystalline polymorph of a coronavirus 3CL protease inhibitor, wherein the coronavirus 3CL protease inhibitor is a compound of formula I, and its crystalline polymorph includes a hydrate pattern A and a 1,4-dioxane solvate pattern B.

[0007]

[0008] The characteristic diffraction peaks of the X-ray powder diffraction pattern of the hydrate Pattern A crystal form, expressed as 2θ value ±0.2°, using Cu-Kα radiation, include: 7.8±0.2°, 13.3±0.2°, 17.0±0.2°, 17.9±0.2°, 19.5±0.2°, 20.2±0.2°, 25.0±0.2°, 26.8±0.2°, 28.2±0.2°, and 30.4±0.2°.

[0009] The differential scanning calorimetry curve of the hydrate Pattern A crystal form showed an endothermic peak at 150.83℃.

[0010] The thermogravimetric analysis curve of the hydrate Pattern A crystal form showed a weight loss of 6.49% between 33℃ and 200℃, and a weight loss of 94.38% at 200.0℃.

[0011] The characteristic diffraction peaks of the X-ray powder diffraction pattern of the 1,4-dioxane solvate Pattern B crystal form, expressed as 2θ values ​​± 0.2°, using Cu-Kα radiation, include: 9.0 ± 0.2°, 14.6 ± 0.2°, 15.5 ± 0.2°, 17.3 ± 0.2°, 20.5 ± 0.2°, 20.7 ± 0.2°, 23.6 ± 0.2°, 24.0 ± 0.2°, and 28.6 ± 0.2°.

[0012] The differential scanning calorimetry (DSC) curves of the 1,4-dioxane solvate Pattern B crystal form showed endothermic peaks at 135.47 °C and 181.38 °C.

[0013] The thermogravimetric analysis curves of the 1,4-dioxane solvate Pattern B crystal form showed a weight loss of 15.737% between 21.51℃ and 150℃, and a weight loss of 2.184% between 100.0℃ and 230.0℃.

[0014] A second aspect of the present invention discloses a method for preparing a crystalline polymorph of the coronavirus 3CL protease inhibitor PLC-01, comprising the following steps:

[0015] Step 1: Dissolve or disperse the compound coronavirus 3CL protease inhibitor in a solvent to obtain a clear solution of coronavirus 3CL protease inhibitor;

[0016] Step 2: Stir the clarified solution of coronavirus 3CL protease inhibitor at 20-50°C to allow crystals to precipitate; or add an antisolvent to the clarified solution of coronavirus 3CL protease inhibitor to allow crystals to precipitate; or slowly evaporate the solvent from the clarified solution of coronavirus 3CL protease inhibitor.

[0017] The solvent is any one of water, methanol, ethanol, acetone, ethyl acetate, isopropyl acetate, acetonitrile, dichloromethane, methyl tert-butyl ether, tetrahydrofuran, or a solvent miscible with water.

[0018] Further, in step 1, the concentration of the clarified solution of the coronavirus 3CL protease inhibitor is 100–400 mg / mL.

[0019] Furthermore, the slow cooling crystallization preparation method for Pattern B crystal form includes the following steps: Weigh the PLC-01 free-state sample, add the minimum volume of solvent and dissolve it completely at 50°C. Pass the resulting solution or dilute suspension through a 0.45μm nylon membrane needle filter to obtain a clear solution. Cool the obtained clear solution to 5°C at a cooling rate of 0.1°C / min and maintain it at 5°C for 1 day. Centrifuge the obtained suspension at 14,000 rpm using a 0.45μm nylon membrane centrifuge tube. The resulting solid is Pattern B crystal form.

[0020] Furthermore, the rapid cooling crystallization preparation method for Pattern B crystal form includes the following steps: Weigh the PLC-01 free-state sample, add a minimum volume of solvent and dissolve it completely at 50°C. Pass the resulting solution or dilute suspension through a 0.45μm nylon membrane needle filter to obtain a clear solution. Place the obtained clear solution in a 0°C ice bath and stir, maintaining it at 0°C for one day. Centrifuge the obtained suspension at 14,000 rpm using a 0.45μm nylon membrane centrifuge tube; the resulting solid is Pattern B crystal form.

[0021] A fourth aspect of the invention discloses a pharmaceutical composition comprising a crystalline polymorph of the aforementioned coronavirus 3CL protease inhibitor, and one or more pharmaceutically acceptable carriers.

[0022] Furthermore, the carrier is selected from fillers, disintegrants, adhesives, lubricants, or combinations thereof.

[0023] The filler is selected from starch, lactose, microcrystalline cellulose, dextrin, mannitol, magnesium oxide, calcium sulfate, or combinations thereof.

[0024] The disintegrant is selected from carboxymethyl cellulose and its salts, croscarmellose and its salts, croscarmellose, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose or combinations thereof.

[0025] The adhesive is selected from polyvinylpyrrolidone, hydroxypropyl methylcellulose, starch paste, or combinations thereof.

[0026] The lubricant is selected from magnesium stearate, calcium stearate, or a combination thereof.

[0027] In a fifth aspect, the present invention discloses the use of the crystalline polymorph of the coronavirus 3CL protease inhibitor in the preparation of medicaments for treating metabolic diseases, tumors, autoimmune diseases, or metastatic diseases.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention provides a crystalline polymorph of a coronavirus 3CL protease inhibitor (PLC-01), comprising Pattern A and Pattern B. Pattern A and Pattern B exhibit excellent chemical and physical stability, effectively resisting the influence of the external environment, ensuring long-term stability of the drug during storage and transportation, and reducing the risk of degradation and transformation. Pattern A demonstrates better solubility, which helps to improve the drug's dissolution rate and bioavailability, thereby enhancing efficacy.

[0030] The method for preparing crystalline polymorphs of coronavirus 3CL protease inhibitor (PLC-01) provided by the present invention is simple and highly controllable. It includes dissolving or dispersing compound PLC-01 in a solvent, obtaining the crystal form by stirring to crystallize, adding antisolvent to precipitate, or slowly evaporating the solvent. The operation is simple, the conditions are mild, and the adaptability is strong. It can be flexibly adjusted according to different solvents, temperatures and reaction conditions, which is convenient for large-scale production.

[0031] Compared with the prior art, the pharmaceutical composition provided by this invention not only optimizes the physicochemical properties of the drug, but also improves the stability and efficacy of the formulation, laying a solid foundation for the further development, clinical application and industrialization of the coronavirus 3CL protease inhibitor (PLC-01) as a potential anti-coronavirus drug. Attached Figure Description

[0032] Figure 1 Ortep image of the free-state Pattern A single crystal structure of the coronavirus 3CL protease inhibitor (PLC-01) of this invention;

[0033] Figure 2This is an X-ray powder diffraction (XRPD) pattern of the free-state Pattern A crystal form of the coronavirus 3CL protease inhibitor (PLC-01) of this invention.

[0034] Figure 3 This is a differential scanning calorimetry (DSC) image of the free-state Pattern A crystal form of PLC-01 in this invention;

[0035] Figure 4 Thermogravimetric analysis (TGA) diagram of the free-state Pattern A crystal form of the coronavirus 3CL protease inhibitor (PLC-01) of this invention;

[0036] Figure 5 This is an X-ray powder diffraction (XRPD) pattern of the free-state Pattern B crystal form of the coronavirus 3CL protease inhibitor (PLC-01) of this invention.

[0037] Figure 6 Differential scanning calorimetry (DSC) curve of the free-state Pattern B crystal form of the coronavirus 3CL protease inhibitor (PLC-01) of this invention;

[0038] Figure 7 The thermogravimetric analysis (TGA) diagram of the free-state Pattern B crystal form of the coronavirus 3CL protease inhibitor (PLC-01) of this invention. Detailed Implementation

[0039] To enable those skilled in the art to understand the features and effects of the present invention, the following descriptions and definitions are only general descriptions of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in the event of any conflict, the definitions in this specification shall prevail.

[0040] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0041] The following examples use instruments and equipment conventional in the art. Experimental methods in the following examples, unless otherwise specified, are generally performed under standard conditions or as recommended by the manufacturer. All raw materials used in the following examples are conventional commercially available products with specifications in the art, unless otherwise stated.

[0042] In this invention, polymorphs refer to the phenomenon where the same chemical substance forms two or more different crystal structures due to different arrangements of molecules or atoms in the crystal. For the compound PLC-01, its crystalline polymorphs include the hydrate Pattern A and the 1,4-dioxane solvate Pattern B. This means that PLC-01 molecules arrange themselves in different ways under different environmental conditions, forming substances with different crystal structures. Polymorphism affects the formulation process of pharmaceuticals. Different polymorphs exhibit different properties such as flowability and compressibility during formulation, which can affect the quality and production efficiency of tablets, capsules, and other formulations.

[0043] The X-ray powder diffraction pattern described in this invention was acquired using a Bruker D8 Advance X-ray powder diffractometer. The parameters of the X-ray powder diffraction method described in this invention are as follows:

[0044] X-ray path: reflection mode; detector: LYNXEYE_XE_T (1D mode); aperture angle: 2.9° (max); radiation source: Cu / K-Alpha1 X-ray source power: 40kV, 40mA; Primary optical path slit: 20.0mm; Primary Solar slit: 2.5°; Secondary optical path slit: 2.5°; Scanning mode: continuous scan; Scanning type: dual optical path mode; Step size: 0.02°; Step time: 0.3s / step; Scanning range: 2° to 40°; Sample rotation speed: 15rpm; Sample plate: single crystal silicon wafer, planar plate.

[0045] The differential scanning calorimetry (DSC) analysis chromatograms described in this invention were acquired on a TA Instruments Discovery 2500. The parameters of the differential scanning calorimetry (DSC) analysis method described in this invention are as follows: temperature range: 30 to 250 °C; heating rate: 10 °C / min; nitrogen flow rate: 50 mL / min; sample volume: 0.5–5 mg.

[0046] The thermogravimetric analysis (TGA) chromatograms described in this invention were acquired on a TA Instruments Discovery 5500. The TGA method parameters described in this invention are as follows: starting temperature: ambient temperature (below 35°C); ending temperature: 300°C; heating rate: 10°C / min; nitrogen flow rate: equilibration 10 mL / min; sample chamber 25 mL / min; sample volume: 2–10 mg.

[0047] The coronavirus 3CL protease inhibitor of this invention is a compound of Formula I, namely (E)-3-((1H-1,2,3-triazol-5-yl)methyl)-6-((6-chloro-2-methyl-2H-indazol-5-yl)imino)-1-(2,4,5-trifluorobenzyl)-1,3,5-triazine-2,4-dione, abbreviated as PLC-01.

[0048]

[0049] PLC-01, as a 3CL protease inhibitor, exhibits strong inhibitory efficacy. Its inhibitory activity reaches the nanomolar level, and its in vitro 3CL protease inhibitory activity is close to that of the marketed drug Ensitrelvir, but it has better pharmacokinetic characteristics.

[0050] Example 1

[0051] This embodiment provides single-crystal data for the PLC-01 free-state Pattern A crystal form, as shown in Table 1. The Ortep diagram of the PLC-01 free-state Pattern A single-crystal structure is shown below. Figure 1 As shown.

[0052] Table 1: Single crystal data of PLC-01 free-state Pattern A crystal form

[0053]

[0054] Example 2

[0055] This embodiment provides the PLC-01 free-state Pattern A crystal form, which is obtained by a suspension preparation method. The specific preparation steps are as follows:

[0056] Weigh approximately 40 mg of PLC-01 free sample, add 0.1–0.4 mL of solvent, and suspend at 25 °C with magnetic stirring at 400 rpm for 2 weeks. Filter the resulting suspension through a 0.45 μm nylon filter centrifuge tube at 14,000 rpm, and characterize the resulting solid fraction using XRPD. Alternatively, weigh approximately 40 mg of PLC-01 free sample, add 0.1–0.4 mL of solvent, and suspend at 50 °C with magnetic stirring at 400 rpm for 1 week. Filter the resulting suspension through a 0.45 μm nylon filter centrifuge tube at 14,000 rpm, and characterize the resulting solid fraction using XRPD.

[0057] The PLC-01 free-state Pattern A crystal form obtained in this embodiment was subjected to Cu-Ka radiation, and the resulting X-ray powder diffraction pattern is shown below. Figure 1 As shown, the diffraction peak parameters are represented by the 2θ angle ± 2° and the interplanar spacing (d), and are shown in Table 2.

[0058] Table 2: Specific diffraction peak parameters of PLC-01 free-state Pattern A crystal form

[0059]

[0060]

[0061] Following the same method, screening was conducted under different conditions. Specific screening conditions are shown in Table 3. The resulting XRPD patterns of the solids are compared with... Figure 2 The consistency proves that the crystal form obtained under these conditions is Pattern A.

[0062] Table 3: Suspension Experiment

[0063]

[0064]

[0065] Example 3

[0066] This embodiment provides the PLC-01 free-state Pattern A crystal form, which is obtained by a room-temperature slow evaporation preparation method. The specific preparation steps are as follows:

[0067] Weigh approximately 20 mg of the PLC-01 free-state sample and dissolve it completely in 1.6–20 mL of solvent. The solution is then filtered through a 0.45 μm nylon needle filter to obtain a clear solution. The resulting clear solution is allowed to slowly evaporate under ambient conditions (approximately 18.2–25.8 °C, 27%–78% RH). The resulting solid fraction is characterized by XRPD. The XRPD plot of the obtained solid is shown below. Figure 2 The consistency proves that the crystal form obtained by screening under this method is Pattern A.

[0068] Following the same method, screening was conducted under different conditions. Specific screening conditions are shown in Table 4. The resulting XRPD patterns of the solids are compared with... Figure 2 The consistency proves that the crystal form obtained under these conditions is Pattern A.

[0069] Table 4: Slow Evaporation Experiment

[0070]

[0071] Example 4

[0072] This embodiment provides the PLC-01 free-state Pattern A crystal form, which is prepared using an anti-solvent dropwise addition method. The specific preparation steps are as follows:

[0073] Weigh approximately 40 mg of the PLC-01 free sample and add 7 mL of solvent to dissolve it completely at ambient temperature (approximately 24-25°C). Pass the resulting solution or dilute suspension through a 0.45 μm nylon needle filter to obtain a clear solution. Slowly add 4 times the volume of antisolvent to the obtained clear solution until a large amount of solid precipitates. Characterize the obtained solid using XRPD. The XRPD plot of the obtained solid is compared with... Figure 2 The consistency proves that the crystal form obtained by screening under this method is Pattern A.

[0074] Following the same method, screening was conducted under different conditions. Specific screening conditions are shown in Table 5. The resulting XRPD patterns of the solids are compared with... Figure 2 The consistency proves that the crystal form obtained under these conditions is Pattern A.

[0075] Table 5: Antisolvent addition test

[0076]

[0077] Example 5

[0078] This embodiment provides the PLC-01 free-state Pattern A crystal form, which is prepared by solvent vapor diffusion crystallization. The specific steps are as follows:

[0079] Approximately 40 mg of PLC-01 free-state sample was weighed and dissolved completely in 0.3 mL of the good solvent dimethyl sulfoxide at ambient temperature (approximately 24-25 °C) using a syringe filter. The resulting solution was filtered through a 0.45 μm nylon membrane to obtain a clear solution. The clear solution was transferred to an 8 mL open-top glass bottle, and then transferred to a 40 mL glass bottle. An anti-solvent was added to the 40 mL bottle. The 40 mL bottle was then tightly sealed and stored at ambient temperature for 30 days. The resulting suspension was centrifuged at 14,000 rpm in a 0.45 μm nylon membrane centrifuge tube. The obtained solid was characterized by XRPD. The XRPD plot of the obtained solid is shown in the figure. Figure 2 The consistency proves that the crystal form obtained by screening under this method is Pattern A.

[0080] Following the same method, screening was conducted under different conditions, as detailed in Table 6. The resulting XRPD patterns of the solids were compared with those of the samples obtained from the samples. Figure 2 The consistency proves that the crystal form obtained under these conditions is Pattern A.

[0081] Table 6: Solvent Diffusion Crystallization Experiment

[0082]

[0083] Example 6

[0084] This embodiment provides the differential thermal analysis (DTA) results of the PLC-01 free-state Pattern A crystal form. A 10 mg sample was placed in an uncovered aluminum crucible (unless otherwise specified). The sample was heated from room temperature to 250 °C at a rate of 10 °C / min under a dry N2 protection environment of 50 mL / min. Simultaneously, the TA software recorded the heat change of the sample during the heating process. Figure 3 As shown, by appendix Figure 3 The data shows that the differential scanning calorimetry curve of the hydrate Pattern A crystal form has an endothermic peak at 150.83℃, which is of great significance for a deeper understanding of the thermal stability and thermal behavior of the PLC-01 free-state Pattern A crystal form.

[0085] Example 7

[0086] This embodiment provides the thermogravimetric analysis (TGA) results of the PLC-01 free-state Pattern A crystal form. A 15 mg sample was placed in a platinum crucible, and segmented high-resolution detection was employed. The heating rate was set to 10 °C / min, and dry nitrogen (N2) gas was introduced at a flow rate of 50 mL / min as a protective gas. The sample was heated from room temperature to 300 °C, and the weight change of the sample during the heating process was recorded using TA software. Figure 4 As shown, the thermogravimetric analysis curve of the hydrate Pattern A crystal form revealed a weight loss of 6.49% between 33℃ and 200℃, with the weight loss being related to the content of water of crystallization in the hydrate; at 200.0℃, the weight loss was 94.38%.

[0087] Example 8

[0088] This embodiment provides the PLC-01 free-state Pattern B crystal form, which is obtained by a slow cooling crystallization method. The specific preparation process is as follows:

[0089] Approximately 40 mg of PLC-01 free-state sample was weighed and dissolved completely in 5 mL of 1,4-dioxane at 50 °C. The resulting solution or dilute suspension was passed through a 0.45 μm nylon needle filter to obtain a clear solution. The clear solution was cooled to 5 °C at a cooling rate of 0.1 °C / min and maintained at 5 °C for 1 day. For samples that did not precipitate solids at 5 °C, crystallization was performed at -20 °C. For samples that did not precipitate solids at -20 °C, antisolvent addition or evaporation experiments were conducted to obtain a suspension. The obtained suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon centrifuge tube. The obtained solid was characterized by XRPD. The obtained PLC-01 free-state Pattern B crystal form was analyzed using Cu-Ka radiation. The resulting X-ray powder diffraction pattern is shown below. Figure 5As shown, the diffraction peak parameters are represented by the 2θ angle ±2 and the interplanar spacing (d), and are shown in Table 7.

[0090] Table 7: Specific diffraction peak parameters of PLC-01 free-state Pattern B crystal form

[0091]

[0092]

[0093] Example 9

[0094] This embodiment provides the PLC-01 free-state Pattern B crystal form, which is obtained using a rapid cooling crystallization preparation method. The specific preparation steps are as follows:

[0095] Approximately 40 mg of PLC-01 free-state sample was weighed and dissolved completely in 5 mL of 1,4-dioxane at 50 °C. The resulting solution or dilute suspension was passed through a 0.45 μm nylon needle filter to obtain a clear solution. The clear solution was placed in an ice bath at 0 °C with stirring and kept at 0 °C for one day. For samples that did not precipitate solids at 0 °C, crystallization was performed at -20 °C. For samples that did not precipitate solids at -20 °C, antisolvent addition or evaporation experiments were conducted. The resulting suspension was centrifuged and filtered at 14,000 rpm using a 0.45 μm nylon centrifuge tube, and the obtained solid was characterized by XRPD. The XRPD plot of the obtained solid was compared with... Figure 5 The consistency proves that the crystal form obtained by screening under this method is Pattern B crystal form.

[0096] Example 10

[0097] This embodiment provides the differential thermal analysis (DTA) results of the PLC-01 free-state Pattern B crystal form. A 10 mg sample was placed in an uncovered aluminum crucible (unless otherwise specified). The sample was heated from room temperature to 250 °C at a rate of 10 °C / min under a dry N2 protection environment of 50 mL / min. Simultaneously, TA software was used to record the heat change of the sample during the heating process. Figure 6 As shown, the differential scanning calorimetry curves of the Pattern B hydrate crystal form showed endothermic peaks at 135.47℃ and 181.38℃.

[0098] Example 11

[0099] This embodiment provides the thermogravimetric analysis (TGA) results of the PLC-01 free-state Pattern B crystal form. A 15 mg sample was placed in a platinum crucible, and a segmented high-resolution detection method was used. The sample was heated from room temperature to 300 °C at a heating rate of 10 °C / min under the protection of dry N2 at a rate of 50 mL / min. Simultaneously, the TA software recorded the weight change of the sample during the heating process. Figure 7 As shown, the thermogravimetric analysis curves of the hydrate Pattern B crystal form revealed a weight loss of 15.737% between 21.51℃ and 150℃, and a weight loss of 2.184% between 100.0℃ and 230.0℃.

[0100] Example 12

[0101] (1) Solid stability test

[0102] This embodiment provides a solid stability evaluation of the PLC-01 free-state Pattern A and Pattern B crystal forms. The specific experimental steps are as follows:

[0103] Open containers containing free-state PLC-01 Pattern A and Pattern B crystal forms were placed at 25℃ / 92%RH, 25℃ / 60%RH, and 40℃ / 75%RH for two weeks, respectively. Closed containers containing free-state PLC-01 Pattern A and Pattern B crystal forms were placed at 60℃ for two weeks. Free-state PLC-01 Pattern A and Pattern B crystal forms were exposed to 1.2 million lux-hours of visible light at 25℃, with experiments conducted using both transparent vials and vials wrapped in aluminum foil. The transparent vials allowed direct light exposure, while the foil-wrapped vials served as controls, examining the effect of light exposure on the samples. The stability of the samples under pressure conditions was analyzed by XRPD and HPLC, and color changes were observed. The experimental results are shown in Tables 8 and 9. It can be observed that the concentration, crystal form, and color of the free-state Pattern A and Pattern B crystal forms of PLC-01 did not change significantly. The results show that both crystal forms exhibit good solid stability under the influence of various factors such as temperature, humidity, and light. The purity, crystal form, and color did not change significantly. This result provides important stability data support for the application of these two crystal forms in related fields.

[0104] Table 8: Solid stability test results of PLC-01 free-state Pattern A crystal form

[0105]

[0106] Table 9: Solid stability test results of PLC-01 free-state Pattern B crystal form

[0107]

[0108]

[0109] (2) Tablet compression experiment

[0110] This implementation provides a PLC-01 free-state Pattern A crystal form simulation tableting experiment, and the specific experimental steps are as follows:

[0111] Approximately 10 mg of PLC-01 free-state Pattern A crystal form was weighed and compressed at 10 MPa for 5 min. The crystal form transformation and changes in crystallinity were studied by XRPD characterization, and the results are shown in Table 10. It was found that the crystal form has good compressibility, making it easy to directly compress into tablets.

[0112] Table 10: Simulated tablet compression experiment

[0113]

[0114] (3) Dry grinding experiment

[0115] This implementation provides a dry grinding experiment simulating the free-state Pattern A crystal form using PLC-01. The specific experimental steps are as follows:

[0116] Approximately 10 mg of PLC-01 free-state Pattern A crystal form was weighed and manually ground for 1, 3, and 5 minutes, respectively. The crystal form transformation and changes in crystallinity were studied by XRPD characterization, and the results are shown in Table 11. It was found that it has good powder properties and is easy to grind.

[0117] Table 11: Simulated tablet compression experiment

[0118]

[0119] (4) Powder property experiment

[0120] This experiment provides a method for testing the physical properties of PLC-01 free-state Pattern A crystal form powder. Approximately 10 mg of PLC-01 free-state Pattern A crystal form was weighed and water or ethanol was added to fully wet the sample. The wetted sample was gently ground with a pestle, and then placed under external environmental conditions to dry for 10 min. XRPD characterization was used to study the changes in crystal form transformation and crystallinity. The specific results are shown in Table 12, revealing that it has good powder properties and is suitable for wet granulation.

[0121] Table 12: Simulated tablet compression experiment

[0122]

[0123] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A crystalline polymorph of a coronavirus 3CL protease inhibitor, characterized in that, The coronavirus 3CL protease inhibitor is a compound as shown in Formula I, which has polymorphic forms including hydrate Pattern A and 1,4-dioxane solvate Pattern B.

2. The crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 1, characterized in that, The characteristic diffraction peaks of the X-ray powder diffraction pattern of the hydrate Pattern A crystal form, expressed as 2θ value ±0.2°, using Cu-Kα radiation, include: 7.8±0.2°, 13.3±0.2°, 17.0±0.2°, 17.9±0.2°, 19.5±0.2°, 20.2±0.2°, 25.0±0.2°, 26.8±0.2°, 28.2±0.2°, and 30.4±0.2°.

3. The crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 2, characterized in that, The differential scanning calorimetry curve of the hydrate Pattern A crystal form showed an endothermic peak at 150.83℃.

4. The crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 2, characterized in that, The thermogravimetric analysis curve of the hydrate Pattern A crystal form showed a weight loss of 6.49% between 33℃ and 200℃, and a weight loss of 94.38% at 200.0℃.

5. The crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 1, characterized in that, The characteristic diffraction peaks of the X-ray powder diffraction pattern of the 1,4-dioxane solvate Pattern B crystal form, expressed as 2θ value ±0.2°, using Cu-Kα radiation, include: 9.0±0.2°, 14.6±0.2°, 15.5±0.2°, 17.3±0.2°, 20.5±0.2°, 20.7±0.2°, 23.6±0.2°, 24.0±0.2°, and 28.6±0.2°.

6. The crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 4, characterized in that, The differential scanning calorimetry (DSC) curves of the 1,4-dioxane solvate Pattern B crystal form showed endothermic peaks at 135.47 °C and 181.38 °C. The thermogravimetric analysis curves of the 1,4-dioxane solvate Pattern B crystal form showed a weight loss of 15.737% between 21.51℃ and 150℃, and a weight loss of 2.184% between 100.0℃ and 230.0℃.

7. A method for preparing the crystalline polymorph of the coronavirus 3CL protease inhibitor according to any one of claims 1 to 6, characterized in that, include Step 1: Dissolve or disperse the compound coronavirus 3CL protease inhibitor in a solvent to obtain a clear solution of coronavirus 3CL protease inhibitor; Step 2: Stir the clarified solution of coronavirus 3CL protease inhibitor at 20-50°C to allow crystals to precipitate; or add an antisolvent to the clarified solution of coronavirus 3CL protease inhibitor to allow crystals to precipitate; or slowly evaporate the solvent from the clarified solution of coronavirus 3CL protease inhibitor.

8. The method for preparing the crystalline polymorph of the coronavirus 3CL protease inhibitor according to claim 7, characterized in that, The solvent is any one of water, methanol, ethanol, acetone, ethyl acetate, isopropyl acetate, acetonitrile, dichloromethane, methyl tert-butyl ether, tetrahydrofuran, or a solvent miscible with water.

9. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises at least one of the crystalline polymorphs of the coronavirus 3CL protease inhibitor according to any one of claims 1 to 5, and a pharmaceutically acceptable carrier.

10. The use of the crystalline polymorph of the coronavirus 3CL protease inhibitor according to any one of claims 1 to 5 in the preparation of medicaments for treating metabolic diseases, tumors, autoimmune diseases or metastatic diseases.

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