Crystal form of compound and use thereof

Abstract

The present application relates to a crystal of 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one, a pharmaceutical composition comprising the crystal, and use thereof.

Description

Compound crystal forms and their applications Technical Field

[0001] This invention belongs to the field of medicinal chemistry. More specifically, this invention relates to a compound crystal form and its applications. Background Technology

[0002] 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one is a gemcitabine prodrug, disclosed in CN111655710B (compound 4), having the following structural formula:

[0003] The compound has been disclosed to have therapeutic activity against tumors and viral infections, and has shown significant safety improvements (improved Therapeutic Index (TI)), particularly in the liver.

[0004] Many compounds exhibit polymorphism. Different crystal forms of the same drug compound may have different melting points and solubilities, thus affecting the drug's overall physicochemical properties, such as processability, thermal stability, storage stability, and dissolution rate, and ultimately its bioavailability and efficacy. However, for a specific drug compound, whether its crystals can be obtained, which crystal form can be obtained, and whether the obtained crystal form is suitable for drug development are usually unpredictable. Summary of the Invention

[0005] In view of the above background, the inventors have provided crystals of 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one in crystal forms 2, 3 and 4, which have improved solubility, high melting point, low hygroscopicity, thermal stability, light stability, excellent processability, dissolution, bioavailability and / or excellent pharmacokinetic or pharmacodynamic properties.

[0006] Therefore, the present invention provides, in one aspect, a crystal of 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one in crystal form 4, which has peaks at the following 2θ diffraction angles in the X-ray powder diffraction pattern: 12.0°±0.2°, 15.2°±0.2°, 15.8°±0.2°, 17.1°±0.2°, 20.9°±0.2°, 22.2°±0.2° and 25.8°±0.2°.

[0007] In one specific embodiment, the crystal of crystal form 4 further has peaks at the following 2θ diffraction angles: 9.8°±0.2°, 14.4°±0.2°, 19.6°±0.2°, 20.1°±0.2°, 24.3°±0.2°, 24.8°±0.2°, 26.4°±0.2°, 32.9°±0.2°, 34.4°±0.2°, and 34.7°±0.2°.

[0008] In one specific embodiment, the X-ray powder diffraction pattern of the crystal of crystal form 4 is substantially consistent with that in Figure 58.

[0009] Another aspect of the invention provides crystals of 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one in crystal form 2, which have peaks at the following 2θ diffraction angles in X-ray powder diffraction patterns: 6.8°±0.2°, 15.1°±0.2°, 16.8°±0.2°, 18.5°±0.2°, 19.5°±0.2°, 20.1°±0.2°, 22.9°±0.2°, 24.7°±0.2°, 27.4°±0.2°, and 29.5°±0.2°.

[0010] In one specific embodiment, the crystal of crystal form 2 further has peaks at the following 2θ diffraction angles: 8.3°±0.2°, 13.6°±0.2°, 14.3°±0.2°, 20.9°±0.2°, 21.5°±0.2°, 26.2°±0.2°, 30.5°±0.2°, 32.2°±0.2°, 33.0°±0.2°, and 37.8°±0.2°.

[0011] In one specific embodiment, the X-ray powder diffraction pattern of the crystal of crystal form 2 is substantially consistent with that in Figure 56.

[0012] Another aspect of the invention provides crystals of 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one in crystal form 3, which have peaks at the following 2θ diffraction angles in X-ray powder diffraction patterns: 8.9°±0.2°, 13.8°±0.2°, 15.5°±0.2°, 16.8°±0.2°, 19.8°±0.2°, 21.4°±0.2°, 22.1°±0.2°, and 27.9°±0.2°.

[0013] In one specific embodiment, the crystal of crystal form 3 further has peaks at the following 2θ diffraction angles: 5.2°±0.2°, 18.1°±0.2°, 23.5°±0.2°, 25.8°±0.2° and 30.1°±0.2°.

[0014] In one specific embodiment, the X-ray powder diffraction pattern of crystal form 3 is substantially consistent with that in Figure 57.

[0015] In this invention, unless otherwise indicated, the X-ray powder diffraction pattern is prepared using Cu Kα radiation.

[0016] Another aspect of the present invention provides a pharmaceutical composition comprising crystals and a pharmaceutical carrier as described in the present invention.

[0017] Another aspect of the present invention provides the use of the crystals described in the invention in the preparation of medicaments for treating tumors.

[0018] In one specific embodiment, the tumor is selected from T-cell lymphoma, soft tissue sarcoma, cholangiocarcinoma, pancreatic cancer, breast cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, non-small cell lung cancer, small cell lung cancer, thymic carcinoma, mesothelioma, renal pelvis carcinoma, ovarian cancer, bladder cancer, urothelial carcinoma, and hepatocellular carcinoma (HCC).

[0019] Another aspect of the present invention provides the use of the crystals and at least one tumor-dissolving agent and / or immunotumor agent according to the present invention in the preparation of a medicament for treating tumors, wherein preferably, the tumor-dissolving agent is selected from 5-fluorouracil, chloroquine, S-1 (i.e., the combination drug tegafur / gemeraxyl / piracetam), vinorelbine, sorafenib, elpamotide, capecitabine, carboplatin, cisplatin, oxaliplatin, aurora kinase inhibitors, EGFR inhibitors, tyrosine kinase inhibitors, topoisomerase inhibitors, nalb-paclitaxel (Nab-PTX), paclitaxel, docetaxel, pemetrexed, curcumin, and radiotherapy; and the immunotumor agent is selected from checkpoint inhibitors, PD-1, PDL1, CTLA-4, and VEGF-A antibodies.

[0020] Another aspect of the present invention provides the use of the crystals described in the invention in the preparation of medicaments for treating viral infections.

[0021] The novel crystal forms (crystal forms 4, 2, and 3) disclosed in this invention have high purity, excellent stability, excellent long-term storage and drug stability, good in vivo exposure after oral administration, moderate half-life, good in vivo pharmacokinetics, and can be used as antitumor and antiviral agents, thus enabling the production of high-quality drugs. Attached Figure Description

[0022] Figure 1 shows the XRPD results of the raw material compound;

[0023] Figure 2 shows the TGA results of the raw material compound;

[0024] Figure 3 shows the DSC results of the raw material compound;

[0025] Figure 4 shows the DVS results of the raw material compound;

[0026] Figure 5 shows the XRPD results of the raw material compound after DVS testing;

[0027] Figure 6 shows the PLM results (10x10) of the raw material compound;

[0028] Figure 7 shows the raw material compounds. 1 H-NMR spectrum;

[0029] Figure 8 shows the XRPD results of the volatilization crystallization of the raw material compound;

[0030] Figure 9 shows the XRPD results of the raw material compound suspension pulping method (room temperature);

[0031] Figure 10 shows the TGA results of the raw material compound suspended in toluene and pulped (at room temperature);

[0032] Figure 11 shows the PLM results of the raw material compound in toluene suspension and pulping method (room temperature);

[0033] Figure 12 shows the XRPD results of the toluene suspension pulping of the raw material compound for 1 week;

[0034] Figure 13 shows the XRPD results of the raw material compound suspension pulping method (50℃);

[0035] Figure 14 shows the XRPD results of the anti-solvent method for the raw material compound;

[0036] Figure 15 shows the XRPD results of the acetone-cyclohexane antisolvent method for the raw material compound;

[0037] Figure 16 shows the PLM results of the acetone-cyclohexane antisolvent method for the raw material compound;

[0038] Figure 17 shows the XRPD results (Form 2) of the raw material compound in dichloromethane suspension pulping scale-up (room temperature);

[0039] Figure 18 shows the XRPD results (Form 3) of the new crystal form of the compound after suspension and pulping in MTBE and toluene (room temperature);

[0040] Figure 19 shows the XRD results of compound Form 2 suspended in H2O and EtOH and pulped (at room temperature) (H2O: Form 4; EtOH: Form 4);

[0041] Figure 20 shows the XRD results of the new crystal form of the compound in n-heptane and cyclohexane (room temperature) after suspension and pulping (n-heptane: Form 2, cyclohexane: Form 2);

[0042] Figure 21 shows the XRD results of the new crystal form of the compound under the antisolvent method (Form 4);

[0043] Figure 22 shows the XRD results of the new crystal form of the compound in methanol and acetone (Form 4);

[0044] Figure 23 is a summary diagram of XRPD for screening compound crystal forms;

[0045] Figure 24 shows the XRPD results of compound suspension pulping (Form 2);

[0046] Figure 25 shows the TGA results (Form 2) of the compound suspended in dichloromethane by pulping method (room temperature);

[0047] Figure 26 shows the DSC results (Form 2) of the compound suspended in dichloromethane by pulping method (room temperature);

[0048] Figure 27 shows the PLM results (Form 2) of the compound in dichloromethane suspension by pulping method (room temperature);

[0049] Figure 28 shows the TGA results (Form 2) of the compound scaled up by pulping in dichloromethane suspension (room temperature);

[0050] Figure 29 shows the DSC results (Form 2) of the compound in dichloromethane suspension and pulping scale-up (room temperature);

[0051] Figure 30 shows the PLM results of the compound in dichloromethane suspension (room temperature) (Form 2);

[0052] Figure 31 shows the new crystal form of the compound. 1 H-NMR spectrum (Form 2);

[0053] Figure 32 shows the TGA results (Form 3) of the new crystal form of the compound suspended in MTBE and pulped (at room temperature);

[0054] Figure 33 shows the DSC results (Form 3) of the new crystal form of the compound suspended in MTBE and pulped (at room temperature);

[0055] Figure 34 shows the PLM results (Form 3) of the new crystal form of the compound in MTBE (room temperature) after suspension and pulping.

[0056] Figure 35 shows the new crystal form of the compound. 1 H-NMR spectrum (Form 3);

[0057] Figure 36 shows the DSC results for compound Form 4;

[0058] Figure 37 shows the TGA results for compound Form 4;

[0059] Figure 38 shows the PLM results of the compound in EtOH (room temperature) after suspension and pulping (Form 4);

[0060] Figure 39 shows the new crystal form of the compound. 1 H-NMR spectrum (Form 4);

[0061] Figure 40 shows the XRD results of the acetone-MTBE system under the antisolvent method (Form 4);

[0062] Figure 41 shows the XRD results of the acetone-n-heptane system under the antisolvent method (Form 4);

[0063] Figure 42 shows the PLM results of volatile crystallization in the compound acetone (Form 4);

[0064] Figure 43 shows the HPLC chromatogram of the compound's stability at 40℃ / 75%RH (Form 1, Zoom 4-40 min);

[0065] Figure 44 shows the HPLC chromatogram of the compound's stability at 60℃ (Form 1, Zoom 5-40 min);

[0066] Figure 45 shows the HPLC chromatogram of the light stability of compound CC2010-3 (Form 1);

[0067] Figure 46 shows the HPLC chromatogram of the compound's stability at 40℃ / 75%RH (Form 2, Zoom 3-40 min);

[0068] Figure 47 shows the HPLC chromatogram of the compound's stability at 60℃ (Form 2);

[0069] Figure 48 shows the HPLC chromatogram of the compound's light stability (Form 2);

[0070] Figure 49 shows the HPLC chromatogram of the compound's stability at 40℃ / 75%RH (Form 4);

[0071] Figure 50 shows the HPLC chromatogram of the compound's stability at 60℃ (Form 4);

[0072] Figure 51 shows the HPLC chromatogram of the compound's light stability (Form 4);

[0073] Figure 52 shows the XRPD plot of the stability of compound Form 1;

[0074] Figure 53 shows the XRPD plot of the stability of compound Form 2;

[0075] Figure 54 shows the XRPD plot of the stability of compound Form 4;

[0076] Figure 55 shows the interconversion relationship of the crystal forms of the compound;

[0077] Figure 56 shows the individual XRPD plot of compound Form 2;

[0078] Figure 57 shows the individual XRPD plot of compound Form 3; and

[0079] Figure 58 shows a single XRPD plot of compound Form 4. Detailed Implementation

[0080] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0081] Unless otherwise indicated, the terms used herein have their general technical meanings as understood by those skilled in the art.

[0082] The starting materials used in the embodiments of this application are known and can be obtained from commercial suppliers, or can be synthesized according to methods known in the art.

[0083] Experimental equipment

[0084] Test instrument parameters

[0085] X-ray powder diffraction (XRPD)

[0086] The equipment used was a Shimadzu XRD-6000, and the sample was scanned using the following parameters:

[0087] The radiation source is a Cu-Kα target.

[0088] The minimum operating voltage and current of the fluorescent tube are 40kV and 30mA, respectively.

[0089] The 2-Theta value of the sample scan range is from 5. o up to 50 o The scanning speed is 5 decimeters per minute.

[0090] Thermogravimetric analysis (TGA)

[0091] Weigh approximately 5 mg of sample into a crucible, protect it with nitrogen, and heat it from 30 °C to 350 °C at a rate of 20 °C / min. Hold the temperature at 350 °C for 1 min.

[0092] Differential Scanning Calorimeter (DSC)

[0093] Weigh approximately 1–5 mg of powder sample and place it in a sealed aluminum crucible. Make a pinhole in the crucible lid. Under nitrogen protection, perform differential thermal scanning by heating from 30°C to 300°C and holding at 300°C for 1 min. The heating rate is 20°C / min.

[0094] Dynamic moisture adsorption (DVS)

[0095] Dynamic moisture adsorption experiments consist of adsorption and desorption. It is generally considered that at a set relative humidity, when the sample weight dm / dt ≦ 0.01%, the adsorption or desorption of moisture by the sample at that relative humidity has reached equilibrium.

[0096] Sample test temperature: T = 25℃;

[0097] Equilibrium time: dm / dt: 0.01% / min; Relative humidity range: 0%~95%~0%; Humidity change per step of RH (%) test: 5%. *At 25±1℃ and 80±2%RH (European Pharmacopoeia 6.0)

[0098] Polarizing microscope (PLM)

[0099] The sample is dispersed in a medium (silicone oil), observed using a 10X eyepiece and a 10X objective lens, and the image is recorded using a camera and computer system.

[0100] High-performance liquid chromatography (HPLC)

[0101] HPLC method for equilibrium solubility of compounds 0.02% Ammonium Acetate: Weigh 0.2g of ammonium acetate into 1000mL of water, dissolve, mix well, and filter through a 0.45μm PVDF membrane to obtain the product.

[0102] Chromatographic conditions for compound stability testing 0.02% Ammonium Acetate: Weigh 0.2g of ammonium acetate into 1000mL of water, dissolve, mix well, and filter through a 0.45μm PVDF membrane to obtain the product.

[0103] Example 1

[0104] Physical characterization of the amorphous compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one)

[0105] 4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one (i.e., compound 4 in Example 1 of CN111655710B, i.e., the starting compound) was prepared and confirmed as a white solid according to the method of Example 1 of CN111655710B.

[0106] The compound was characterized by physicochemical methods including XRPD, TGA, DSC, and DVS.

[0107] XRPD characterization results showed that the compound was amorphous (Figure 1). TGA showed a significant weight loss step from room temperature (RT) to 130°C, with a weight loss of 3.47%, indicating that the compound may contain a large amount of residual solvent or moisture. DVS showed that the compound was hygroscopic, with a hygroscopicity of 6.01% at 85% RH, indicating that it is easily hygroscopic (Figures 2 to 6). After DVS testing, the crystal form remained amorphous. 1 The H-NMR spectrum is shown in Figure 7.

[0108] Example 2

[0109] Solubility test of amorphous compounds

[0110] To select a suitable method for crystal form screening, approximate solubility tests were performed on this compound. The solvent list is shown in Table 1. Test method: Approximately 3 mg of the compound was weighed, and appropriate volumes of solvent were added gradually. The mixture was shaken, and the dissolution was observed. The volume of completely dissolved solvent and the volume of solvent that did not completely dissolve previously were recorded. The total amount of solvent with very poor solubility added should not exceed 3 mL. The approximate solubility was calculated.

[0111] Based on approximate solubility results, this amorphous compound is soluble in MeOH, EtOH, IPA, ACN, Acetone, MEK, EtOAc, THF, DMSO, 1,4-Dioxane, and mixed solvents; slightly soluble in water and MTBE; and extremely slightly soluble or insoluble in other media. Unless otherwise specified, all mixed solvent compositions are volume ratios.

[0112] Table 1

[0113] Example 3

[0114] Crystal form screening

[0115] Different methods were used to screen the crystal forms of the above compounds, and the initial amorphous morphology was defined as Form 1 (amorphous).

[0116] 3.1 Compound Crystal Form Screening - Volatile Crystallization Method

[0117] Approximately 40 mg of the compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed into a glass bottle, and a good solvent was added to completely dissolve the compound. The sample bottle was then capped with perforated aluminum foil and placed in a fume hood to allow the solvent to evaporate naturally. The solid was collected and its XRPD was measured. If the measured XRPD pattern did not match the crystal form of the starting drug substance, further characterization studies, such as TGA and DSC, were performed. Specific phenomena and processing procedures are shown in Table 2 below.

[0118] Since the initial compound was in an amorphous state and had high solubility, evaporation crystallization was used as the first screening method for many solvents. XRPD results (Figure 8) showed that the solids obtained in various solvents were all amorphous or oily, with no recrystallization and no new crystal forms formed. This may be related to the still relatively high supersaturation in the solution, resulting in the precipitation of amorphous and oily substances instead of crystals.

[0119] Table 2

[0120] 3.2 Compound Crystal Form Screening – Suspension Slurry Method (Room Temperature)

[0121] Approximately 40 mg of the compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed into a glass vial, and then 0.2–1.0 mL of solvent was added. The selected solvent and sample weight are shown in Table 3. All samples were stirred at room temperature for 3 days. The resulting suspension was centrifuged (12000 rpm, 5 min) to collect the wet solid. If no solid precipitated or only a small amount precipitated, the solid was dried under a fume hood. The collected solid was further dried under reduced pressure overnight, and then the XRPD of the dried solid was measured. If the measured XRPD differed from the initial crystal form Form 1, further studies were conducted using TGA and DSC to confirm the new crystal form.

[0122] XRPD results showed that the crystal form prepared by the room temperature pulping method (Figure 9) was amorphous or oily, except in dichloromethane. This may be related to the low solubility of the compound in these solvents (except dichloromethane), which does not provide sufficient solubility to support its crystal transformation, thus remaining amorphous and oily. The crystal form of the compound appearing in dichloromethane is named Form 2.

[0123] Compound crystal form screening was performed by toluene suspension slurrying (1 week). The slurrying time was increased to observe whether the compound crystallized. Approximately 40 mg of the compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed into a glass vial, and then 1 mL of toluene solvent was added. The sample was stirred at room temperature for 1 week, and the resulting suspension was centrifuged (12000 rpm, 5 min) to collect the wet solid. The collected solid was further vacuum dried overnight in a vacuum drying oven under reduced pressure. The XRPD of the dried solid was then measured. If the measured XRPD differed from the initial crystal form (Form 1), further tests such as TGA and DSC were performed to confirm the new crystal form. The XRPD results of the sample suspended in toluene for one week were consistent with those of the 3-day result. See Figures 10 to 12.

[0124] Table 3

[0125] Similarly, the suspension pulping method was carried out at 50°C for crystal form screening. The XRPD results showed that all the obtained solids were amorphous or had poor crystal form (see Table 4 and Figure 13).

[0126] Table 4

[0127] 3.3 Compound Crystal Form Screening – Antisolvent Method

[0128] Approximately 30 mg of the compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed into a glass vial. An appropriate volume of good solvent was added to completely dissolve the compound. Then, different types of antisolvents were added dropwise until sufficient solid precipitated. If no solid precipitated, the addition of antisolvents was stopped. All samples were stirred at room temperature for 2 days, and the wet solid was collected by centrifugation. The collected solid was further dried under reduced pressure overnight, and the XRPD of the dried solid was then measured. If the measured XRPD pattern did not match the crystal form of the starting drug substance, further characterization studies, such as TGA and DSC, were performed.

[0129] Since the samples were all clear solutions, they were evaporated to dryness before XRPD testing. The results are shown in Table 5 and Figures 14 to 16.

[0130] Table 5

[0131] 3.4 Compound Crystal Form Screening – Using Form 2 New Crystal Forms as Raw Materials

[0132] The initial compound Form 1 (amorphous) did not yield good crystal form screening results in the first round of crystal form screening due to various possible factors. Therefore, Form 2 was selected as the raw material for the second round of crystal form screening.

[0133] Preparation of Compound Form 2: Form 2 was prepared by slurrying the initial compound (Form 1, amorphous form) in dichloromethane. Approximately 400 mg of the compound was weighed into a 40 mL glass bottle, and then 5 mL of dichloromethane solvent was added. The sample was stirred at room temperature for 3 days. The resulting suspension was centrifuged (12000 rpm, 5 min) to collect the wet solid. The collected solid was further vacuum dried overnight in a vacuum drying oven under reduced pressure. The XRPD of the dried solid was then measured to confirm consistency with the small-scale test. Specifically, 402.56 mg of the sample was weighed, and after slurrying, Form 2 (363.41 mg) was obtained, with a yield of 90.21%. The obtained sample is Form 2. See Figure 17.

[0134] 3.4.1 Compound Crystal Form Screening (Form 2 Raw Materials) Suspension Pulping

[0135] Approximately 25 mg of the new crystalline form Form 2 was weighed into a glass vial, and then 0.5 mL of solvent was added, along with the selected solvent and sample volume. All samples were stirred at room temperature for 3 days. The resulting suspension was centrifuged (12000 rpm, 5 min) to collect the wet solid. The collected solid was further vacuum dried overnight in a vacuum drying oven under reduced pressure. The XRPD of the dried solid was then measured. If the measured XRPD differed from that of the initial crystalline form Form 2, further tests such as TGA and DSC were conducted to confirm the new crystalline form. The results are shown in Table 6 and Figures 18 to 20, yielding new crystalline forms 3 (Form 3) and 4 (Form 4).

[0136] Table 6

[0137] 3.4.2 Compound Crystal Form Screening (Form 2 Raw Materials) Antisolvent Method

[0138] Approximately 25 mg of Form 2 was weighed into a glass vial, and an appropriate volume of good solvent was added to completely dissolve it. Then, different types of antisolvents were added dropwise until sufficient solid precipitated. If no solid precipitated, the addition of antisolvents was stopped. All samples were stirred at room temperature for 2 days, and the wet solid was collected by centrifugation. The collected solid was further dried under reduced pressure overnight, and the XRPD of the dried solid was then measured. If the measured XRPD spectrum did not match the crystal form of the starting drug substance, further characterization studies, such as TGA and DSC, were performed. The results are shown in Table 7 and Figure 21, yielding crystal form 4.

[0139] Table 7

[0140] 3.4.3 Compound Crystal Form Screening (Form 2 Raw Materials) Volatile Crystallization Method

[0141] Weigh approximately 25 mg of Form 2 into a glass bottle and add a good solvent to completely dissolve the compound. Then, cover the bottle with perforated aluminum foil and place it in a fume hood to allow the solvent to evaporate naturally. Collect the solid and measure its XRPD. If the measured XRPD pattern does not match the crystal form of the starting drug substance, further characterization studies, such as TGA and DSC, will be performed. Specific phenomena and processing procedures are shown in the table below. The results are shown in Table 8 and Figure 22, yielding crystal form 4.

[0142] Table 8

[0143] Example 4

[0144] Characterization of new crystal forms of compounds

[0145] As described above, in the polymorph screening study of compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one), three new polymorphs were discovered: polymorph 2, polymorph 3, and polymorph 4. The new polymorphs were characterized. The results are shown in Table 9 and Figures 23 to 42. The individual XRPD spectra of polymorphs 2, 3, and 4 are shown in Figures 56 to 58, respectively.

[0146] In the XRPD spectrum of crystal form 4, the main significant peaks include those at the following 2θ diffraction angles: 9.8°±0.2°, 12.0°±0.2°, 14.4°±0.2°, 15.2°±0.2°, 15.8°±0.2°, 17.1°±0.2°, 19.6°±0.2°, 20.1°±0.2°, 20.9°±0.2°, 22.2°±0.2°, 24.3°±0.2°, 24.8°±0.2°, 25.8°±0.2°, 26.4°±0.2°, 32.9°±0.2°, 34.4°±0.2°, and 34.7°±0.2°.

[0147] In the XRPD spectrum of crystal form 2, the main significant peaks include those at the following 2θ diffraction angles: 6.8°±0.2°, 8.3°±0.2°, 13.6°±0.2°, 14.3°±0.2°, 15.1°±0.2°, 16.8°±0.2°, 18.5°±0.2°, 19.5°±0.2°, 20.1°±0.2°, 20.9°±0.2°, 21.5°±0.2°, 22.9°±0.2°, 24.7°±0.2°, 26.2°±0.2°, 27.4°±0.2°, 29.5°±0.2°, 30.5°±0.2°, 32.2°±0.2°, 33.0°±0.2°, and 37.8°±0.2°.

[0148] In the XRPD spectrum of crystal form 3, the main significant peaks include those at the following 2θ diffraction angles: 5.2°±0.2°, 8.9°±0.2°, 13.8°±0.2°, 15.5°±0.2°, 16.8°±0.2°, 18.1°±0.2°, 19.8°±0.2°, 21.4°±0.2°, 22.1°±0.2°, 23.5°±0.2°, 25.8°±0.2°, 27.9°±0.2°, and 30.1°±0.2°.

[0149] Example 5

[0150] Preparation of crystal forms 4 and 2 of compounds

[0151] An attempt was made to prepare Form 4 using the active pharmaceutical ingredient (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) in ethanol. Approximately 100 mg of the compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed into a liquid chromatography vial, and then 3 mL of ethanol was added. It was found that the dissolution did not occur as in the small-scale test; the compound did not dissolve after the addition of ethanol. Therefore, a slurry method was chosen. All samples were stirred at room temperature for 3 days. The resulting suspension was centrifuged (12000 rpm, 5 min) to collect the wet solid. The collected solid was then further vacuum dried overnight in a vacuum drying oven under reduced pressure. The XRPD of the dried solid was then measured. If the measured XRPD differed from the initial amorphous form (Form 1), further tests such as TGA and DSC were conducted to confirm the new crystal form. The results are shown in Table 10. The XRPD test results were consistent with those of Form 4.

[0152] Table 10

[0153] The preparation of compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidine-2(1H)-one) crystal form 2 was as described in Example 3.

[0154] Example 6

[0155] Solid stability evaluation of candidate crystal forms

[0156] Amorphous samples of compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) in Forms 1, 2, and 4 were placed in a 60°C-closed, 40°C / 75%RH-open environment with a light intensity of 4500 Lx ± 500 Lx and a total illuminance not less than 1.2 × 10⁶ Lux·hr, and a near-UV lamp intensity not less than 200 W·hr / m². 2 Under the conditions of ), the changes in crystal form and impurities were examined at 1 week and 2 weeks respectively.

[0157] Different forms of the compounds were tested under 60℃, 40℃ / 75%RH, and light conditions. Form 1 showed a significant increase in impurities under 40℃ / 75%RH and 60℃ conditions for two weeks, but good stability under light conditions. Form 2 showed significant impurities under 40℃ / 75%RH conditions for two weeks, but good stability under 60℃ and light conditions, with no significant increase in impurities. Form 4 showed good stability under all three conditions, with a slight increase in impurities. The crystal forms of Forms 1, 2, and 4 did not change significantly during the physical stability tests. The results are shown in Table 11 and Figures 43 to 54.

[0158] Sample preparation method: Weigh 5 mg of sample into a clean glass bottle, add methanol and sonicate until completely dissolved, then analyze by HPLC.

[0159] Example 7

[0160] Solubility test of candidate compound crystals

[0161] Approximately 5 mg of each of the amorphous Forms 1, 2, and 4 of compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) was weighed and added to 1 mL of water, pH 1.0, pH 4.5, and pH 6.8 buffer solutions, respectively. The samples were then magnetically stirred at 37°C and 200 rpm. After 24 hours, the samples were collected, centrifuged at 12000 rpm for 5 min, and the supernatant was diluted with methanol to determine the content. The solubility results show that at 37℃, the solubility of the amorphous form of Form 1 is relatively greater than that of the crystalline forms Form 2 and Form 4, while the solubility of Form 2 is also greater than that of Form 4. See Table 12.

[0162] Table 12

[0163] Summarize

[0164] (1) Physicochemical characterization of the starting compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one) (see Figures 1 to 6): XRPD showed only diffuse broad peaks without obvious diffraction peaks, indicating that the starting compound was an amorphous compound, defined as Form 1 (amorphous). TGA showed a significant weight loss step from room temperature (RT) to 130 °C, with a weight loss of 3.47%, indicating that the compound may contain a large amount of solvent residue or moisture. DVS showed that the compound was hygroscopic, with a hygroscopicity of 6.01% at 85% RH. Based on the approximate solubility results, the starting compound is soluble in MeOH, EtOH, IPA, ACN, Acetone, MEK, EtOAc, THF, DMSO, 1,4-Dioxane, and mixed solvents, but has poor solubility in other media.

[0165] (2) The first round of crystal form screening was performed on the starting compound (4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one). Commonly used crystal form screening methods include pulping, solvent evaporation, antisolvent method, cooling crystallization, and heating method. In the preparation of crystal forms by room temperature pulping, XRPD results showed that the crystal form obtained in dichloromethane was different from the initial crystal form, defined as Form 2 (crystal form). TGA and DSC showed a weight loss of 1.13% from room temperature (RT) to 120℃ and a melting point of 128℃. The crystal forms obtained by other solvents were consistent with the initial crystal form or were of poor quality. Due to the high supersaturation of the amorphous compound, satisfactory results were not obtained. Therefore, the starting material for crystal form screening was changed to Form 2 for a second round of crystal form screening. Form 2 was slurried in different solvents. In water and ethanol, a new crystal form (Form 4) was obtained, which was anhydrous and had good crystallinity. TGA and DSC showed a weight loss of 0.17% from room temperature (RT) to 120℃, and a melting point of 193℃. In MTBE, another crystal form (Form 3) was obtained, presumably a solvate. Antisolvent testing was conducted using Form 2 as the starting material. In the acetone-methyl tert-butyl ether system, the solid obtained was always Form 4. In the volatile crystallization test, Form 4 was also obtained in acetone. The interconversion relationships of crystal forms are shown in Figure 55.

[0166] (3) Form 2 and Form 4 were selected as candidate crystal forms, and repeated preparation experiments were conducted on the new crystal forms Form 2 and Form 4. Form 2 could be successfully scaled up by slurrying in dichloromethane with a high yield (90%). TGA and DSC showed a weight loss of 0.79% from room temperature (RT) to 120℃ and a melting point of 131℃. An attempt was made to obtain Form 4 in one step using CC2010-3 amorphous raw material in ethanol.

[0167] (4) The three different forms of compound CC2010-3 (Form 1, Form 2, and Form 4) were evaluated to select the most suitable crystal form for development. The stability of compound CC2010-3 under 60℃, 40℃ / 75%RH, and light conditions was evaluated. Form 1 (active drug substance, amorphous) showed a significant increase in impurities under 40℃ / 75%RH and 60℃ conditions for 2 weeks, but good stability under light conditions. Form 2 showed significant impurities under 40℃ / 75%RH conditions for 2 weeks, but good stability under 60℃ and light conditions, with no significant increase in impurities. Form 4 showed good stability under all conditions, with a slight increase in impurities. The physical stability of Form 1, Form 2, and Form 4 did not show significant changes in crystal form. Solubility was evaluated by comparing the equilibrium solubility in water and buffer solutions at pH 1.0, pH 4.5, and pH 6.8 at 37℃. The results showed that at 37℃, the order of solubility was amorphous Form 1 > crystalline Form 2 > crystalline Form 4. The equilibrium solubility of Form 4 in these four solvents ranged from 0.12 to 0.19 mg / mL.

[0168] In summary, comparing the above-mentioned morphologies (or crystal forms) of the compounds, Form 4 has a high melting point, good potential thermal stability, no hygroscopicity, good stability, acceptable solubility, and is simple to prepare by recrystallization. Therefore, it is the most suitable for drug development.

[0169] Example 8

[0170] Pharmacokinetic study of crystal form 4 of compound

[0171] The pharmacokinetic distribution of the above compound in plasma was determined by LC-MS / MS after a single oral gavage administration to rats of crystal form 4.

[0172] Test principle: The plasma concentration of the target drug at different time points was determined by LC-MS / MS to investigate the pharmacokinetic characteristics of rats after a single oral administration of 10 mpk (drug mass mg / animal weight kg).

[0173] Experimental Methods: An appropriate amount of the compound in crystal form 4 was accurately weighed and dissolved in a solution of 40% sulfobutyl-β-cyclodextrin (SBE-β-CD): 10% vitamin E polyethylene glycol succinate (TPGS) = 50:50 (V / V) to obtain a dosage form with a concentration of 1 mg / mL. Sprague-Dawley rats / SPF (Shanghai Experimental Animal Center) were used. A total of 12 rats (6 males / 6 females) were administered the drug via gavage at a dose of 10 mpk. Blood samples were collected at 0.083 hr, 0.25 hr, 0.5 hr, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr, 32 hr, and 48 hr (N ​​= 3 / time point). Approximately 0.4 mL of whole blood (100 μL of 10.0 mg / mL tetrahydrouridine anticoagulation) was collected at each time point and stored on wet ice. The stabilizer-to-whole blood ratio was 1:10. Within 60 minutes of blood collection, red blood cells were separated by centrifugation at 2000g for 10 minutes at 4°C to obtain plasma samples. The samples were transferred to two cryovials (approximately 80 μL each). After centrifugation, the plasma samples were transferred to wet ice and stored at -75±15°C until analysis. Liquid chromatography-mass spectrometry (LC-MS) was used, with Verapamil as an internal standard, to determine the concentrations of compounds, dFdC, and dFdU in SD rat plasma. Mass spectrometry analysis was performed using a Triple Quad 5500+ mass spectrometer in positive ion mode with an electrospray ionization source.

[0174] Based on drug concentration-time data, Phoenix was used. TM WinNonlin version 8.3 software calculates pharmacokinetic parameters using a non-compartmental model, including peak concentration (Cmax), time to peak concentration (Tmax), area under the drug-time curve (AUC), and elimination half-life (t). 1 / 2 The AUC is calculated using the linear trapezoidal rule (linear up log down).

[0175] Table 13: Pharmacokinetic parameters of compounds NC: This value cannot be calculated using WinNonlin.

[0176] The experimental results show that the compound of the present invention, crystal form 4, has good in vivo exposure after oral administration, moderate half-life, good in vivo pharmacokinetics, and has the potential to become a drug.

[0177] Example 9

[0178] Pharmacodynamic evaluation of compound crystal form 4

[0179] 1. Experimental Objective: To evaluate the antitumor effect of the test drug on a subcutaneous xenograft tumor model of human non-small cell lung cancer NCI-H1975 cells.

[0180] 2. Test methods:

[0181] 2.1 Cell Culture:

[0182] Human lung cancer NCI-H1975 cells (ATCC, catalog number: CRL-5908) were cultured in vitro in a monolayer under the following conditions: RPMI 1640 medium supplemented with a mixture of 10% fetal bovine serum and 1% penicillin-streptomycin, and incubated at 37°C in a 5% CO2 incubator. Cells were passaged twice a week using trypsin-EDTA digestion. When the cells reached the exponential growth phase, they were harvested, counted, and seeded.

[0183] 2.2 Animals: BALB / c mice, female, 6-8 weeks old, weighing 18-24 grams. Provided by Shanghai Xipu-Bikai Laboratory Animal Co., Ltd. or other suppliers.

[0184] 2.3 Tumor inoculation and animal grouping: 0.1 mL (5 × 10⁻⁶) of the drug was administered to the tumor. 6 NCI-H1975 cells were subcutaneously inoculated into the right posterior dorsal region of each nude mouse. On day 10 post-inoculation, the average tumor volume reached approximately 143 mm². 3 Treatment began at that time. Mice were randomly assigned to groups based on tumor volume using the Astra software. Each group contained 8 tumor-bearing mice.

[0185] The control group was given the following solvent: 40g of sulfobutyl-β-cyclodextrin was weighed into a reagent bottle, and an appropriate amount of deionized water was added. The mixture was stirred, vortexed, and sonicated as needed, and finally brought to a final volume of 100mL to obtain 40% SBE-β-CD. 10g of vitamin E polyethylene glycol succinate was weighed, and an appropriate amount of deionized water was added. The mixture was stirred, vortexed, and sonicated as needed, and finally brought to a final volume of 100mL to obtain 10% TPGS. Equal volumes of 40% SBE-β-CD and 10% TPGS were added to a suitable container and stirred thoroughly to obtain a 40% SBE-β-CD:10% TPGS ratio of 50:50 (v / v).

[0186] The experimental group received a dose of 15 mg / kg, administered orally via gavage at a volume of 10 μl / g based on the mouse's body weight, once daily. The animals' health was monitored daily; if the tumor volume exceeded 3,000 mm, [further action was taken]. 3 If a person has a serious illness, pain, or a weight loss of more than 20% that continues to worsen, euthanasia is necessary.

[0187] 2.4 Tumor growth inhibition rate: Tumor diameter was measured three times a week using calipers. The formula for calculating tumor volume is: V = 0.5a × b 2, where a and b represent the long and short diameters of the tumor, respectively. The formula for calculating the tumor-suppressive efficacy (TGI%) of the compound in each group is: TGI(%) = [1 - (Ti - T0) / (Vi - V0)] × 100, where Ti is the average tumor volume at the end of treatment in a certain treatment group, T0 is the average tumor volume at the beginning of treatment in that treatment group, Vi is the average tumor volume at the end of treatment in the solvent control group, and V0 is the average tumor volume at the beginning of treatment in the solvent control group. One-way ANOVA was used to compare tumor volume and tumor weight among the groups. A significant difference in the F-value (the ratio of treatment variance to error variance) was tested using the Games-Howell test. All data were analyzed using Prism 10.0. A p-value < 0.05 was considered statistically significant.

[0188] 3. Experimental results: According to this model, the crystal form 4 compound of the present invention can significantly inhibit tumor growth (TGI(%) = 94%), which is statistically significant compared with the control group (p < 0.05), and has no inhibitory effect on mouse body weight, indicating good safety.

[0189] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0190] It should also be noted that the various specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Furthermore, various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the spirit of the present invention, and should also be regarded as the content disclosed by the present invention.

Claims

Crystallization 4 of 1,4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one, which has peaks at the following 2θ diffraction angles in its X-ray powder diffraction pattern: 12.0°±0.2°, 15.2°±0.2°, 15.8°±0.2°, 17.1°±0.2°, 20.9°±0.2°, 22.2°±0.2° and 25.8°±0.2°.

2. The crystal according to claim 1, wherein the crystal further has peaks at the following 2θ diffraction angles: 9.8°±0.2°, 14.4°±0.2°, 19.6°±0.2°, 20.1°±0.2°, 24.3°±0.2°, 24.8°±0.2°, 26.4°±0.2°, 32.9°±0.2°, 34.4°±0.2°, and 34.7°±0.2°.

3. The crystal according to claim 1 or 2, wherein the X-ray powder diffraction pattern is substantially consistent with FIG.

58. Crystal form 2 of 4,4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one, which has peaks at the following 2θ diffraction angles in its X-ray powder diffraction pattern: 6.8°±0.2°, 15.1°±0.2°, 16.8°±0.2°, 18.5°±0.2°, 19.5°±0.2°, 20.1°±0.2°, 22.9°±0.2°, 24.7°±0.2°, 27.4°±0.2° and 29.5°±0.2°.

5. The crystal according to claim 4, wherein the crystal further has peaks at the following 2θ diffraction angles: 8.3°±0.2°, 13.6°±0.2°, 14.3°±0.2°, 20.9°±0.2°, 21.5°±0.2°, 26.2°±0.2°, 30.5°±0.2°, 32.2°±0.2°, 33.0°±0.2°, and 37.8°±0.2°.

6. The crystal according to claim 4 or 5, wherein the X-ray powder diffraction pattern is substantially consistent with FIG.

56. 7.4-((2R,4S)-4-(3-chlorophenyl)-2-oxo-1,3,2-dioxophosphoryl-2-amino)-1-(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one crystal of crystal form 3, which has peaks at the following 2θ diffraction angles in the X-ray powder diffraction pattern: 8.9°±0.2°, 13.8°±0.2°, 15.5°±0.2°, 16.8°±0.2°, 19.8°±0.2°, 21.4°±0.2°, 22.1°±0.2° and 27.9°±0.2°.

8. The crystal according to claim 7, wherein the crystal further has peaks at the following 2θ diffraction angles: 5.2°±0.2°, 18.1°±0.2°, 23.5°±0.2°, 25.8°±0.2° and 30.1°±0.2°.

9. The crystal according to claim 7 or 8, wherein the X-ray powder diffraction pattern is substantially consistent with FIG.

57.

10. The crystal according to any one of claims 1 to 9, wherein the X-ray powder diffraction pattern is prepared using Cu Kα radiation.

11. A pharmaceutical composition comprising crystals and a pharmaceutical carrier according to any one of claims 1 to 10.

12. Use of the crystal according to any one of claims 1 to 10 in the preparation of a medicament for treating tumors.

13. The application according to claim 12, wherein the tumor is selected from T-cell lymphoma, soft tissue sarcoma, cholangiocarcinoma, pancreatic cancer, breast cancer, Hodgkin lymphoma, non-Hodgkin lymphoma, non-small cell lung cancer, small cell lung cancer, thymic carcinoma, mesothelioma, renal pelvis carcinoma, ovarian cancer, bladder cancer, urothelial carcinoma, and hepatocellular carcinoma (HCC).

14. The use of the crystal and at least one tumor-dissolving agent and / or immunotumor agent according to any one of claims 1 to 10 in the preparation of a medicament for treating tumors, wherein preferably, the tumor-dissolving agent is selected from 5-fluorouracil, chloroquine, S-1 (i.e., the combination drug tegafur / gemeracil / piracetam), vinorelbine, sorafenib, elpamotide, capecitabine, carboplatin, cisplatin, oxaliplatin, aurora kinase inhibitors, EGFR inhibitors, tyrosine kinase inhibitors, topoisomerase inhibitors, nalbu-paclitaxel, paclitaxel, docetaxel, pemetrexed, curcumin, and radiotherapy; and the immunotumor agent is selected from checkpoint inhibitors, PD-1, PDL1, CTLA-4, and VEGF-A antibodies.

15. Use of the crystal according to any one of claims 1 to 10 in the preparation of a medicament for treating viral infections.

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