Crystalline form of compound yzg-331 and process for preparation thereof

Abstract

This invention belongs to the field of pharmaceutical technology and discloses a new crystal form and preparation method of YZG-331, a candidate compound for treating insomnia, as shown in the following formula. Specifically, the preparation methods and characteristics of three crystal forms of compound YZG-331 are disclosed.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical crystal technology. Specifically, it relates to a novel crystal form of compound YZG-331 and its preparation method. Background Technology

[0002] In light of the traditional application of Gastrodia elata and other Chinese medicinal herbs primarily through decoction, the project team conducted research on the water extract of Gastrodia elata. From this extract, a series of components, including p-hydroxybenzyl alcohol phenols, p-hydroxybenzyl alcohol glycosides, and benzyl citrate ester glycosides, have been isolated and identified [1-10]. A trace component, N, was also discovered. 6 -(p-hydroxybenzyl)-adenosine (NHBA), with a yield of approximately 0.0004%, exhibits a very significant sedative effect; when administered intraperitoneally, its sedative effect is more than 1,000 times that of components such as gastrodin [11,12]. Given the low oral bioavailability of NHBA (the effective dose differs by more than 200 times between gavage and intraperitoneal administration), based on the confirmation that NHBA is a key pharmacodynamic component for the sedative-hypnotic effect of Gastrodia elata, we further conducted repeated synthesis and pharmacodynamic, preliminary drug metabolism, and toxicity tests to evaluate its drug-likeness, ultimately obtaining the sedative-hypnotic candidate drug YZG-331. Furthermore, we investigated the N... 6 -Adenosine derivatives and N 6 - The substituted purine derivatives, their preparation methods, pharmaceutical compositions and uses, as well as their tablet dosage forms and preparation methods have been patented [13-17] and have been granted patents in China and the United States. In order to investigate the potential crystal forms of YZG-331, a polymorph screening experiment was carried out, and three crystal forms of YZG-331 were obtained.

[0003] References:

[0004] 1. Wang Yanan. Study on chemical constituents and bioactivity of Gastrodia elata [D]. Peking Union Medical College, 2012.

[0005] 2. Guo Qinglan. Study on chemical constituents and bioactivity of Gastrodia elata [D]. Peking Union Medical College, 2014.

[0006] 3. Zhou Xue. Study on chemical composition of Gastrodia elata water extract [D]. Peking Union Medical College, 2017.

[0007] 4. Wang Yanan, Lin Sheng; Chen Minghua; et al. Chemical constituents of Gastrodia elata aqueous extract [J]. Chinese Journal of Traditional Chinese Medicine 2012, 37: 1775. 5. Guo QL; Wang YN; Zhu CG; et al. 4-Hydroxybenzyl-substituted glutathione derivatives from Gastrodia elata [J]. J. Asian Nat. Prod. Res. 2015, 17: 439.

[0008] 6.Guo Q.; Wang Y.; Lin S.; et al.4-Hydroxybenzyl-substituted amino acidderivatives from

[0009] Gastrodia elata[J].Acta Pharm.Sin.B 2015,5:350.

[0010] 7. Guo QL; Lin S.; Wang YN; et al. Gastrolatathioneine, an unusualergothioneine derivative from an aqueous extract of “tian ma”: A natural product co-produced by plant and symbioticfungus. Chin. Chem. Lett. 2016, 27: 1577.

[0011] 8.Zhou

[0012] Chin.Chem.Lett.2017,28:1185.

[0013] 9.Xu CB; Guo QL; Wang YN; et al.Gastrodin derivatives from Gastrodia elata[J].Nat.Prod.

[0014] Bioprospect. 2019, 9:393.

[0015] 10.Wang Y.; Zhang M.; Zhou X.; et al. Insight into medicinal chemistry behind traditional Chinese medicines: p-hydroxybenzyl alcohol-derived dimers and trimers from Gastrodia elata[J].Nat.

[0016] Prod.Bioprospect.2021,11:31.

[0017] 11. Zhang Y.; Li M.; Kang RX; at al.NHBA isolated from Gastrodia elataexerts sedative and hypnotic effects in sodium pentobarbital-treated mice[J].Pharm.Biochem.Behav.2012,102:450.

[0018] 12.He J.; Luo Z.; Huang L.; et al.Ambient mass spectrometry imagingmetabolomics method provides novel insights into the action mechanism of drugcandidates[J]. Anal.Chem.2015,87:5372.

[0019] 13. Shi Jiangong, Zhang Jianjun, Yue Zhenggang, et al. N 6 -Substituted adenosine derivatives and N 6 -Substituted adenine derivatives and their uses [P]. ZL200980162826.5.

[0020] 14. Zhang Jianjun, Shi Jiangong, Li Min, et al. N 6 -Substituted adenosine derivatives, their preparation, pharmaceutical compositions and uses [p]. ZL 200810114594.3.

[0021] 15. Shi Jiangong, Zhang Jianjun, Yue Zhenggang, et al. N 6 -Substituted adenosine derivatives and N 6-Substituted adenine derivatives and their uses [p]. ZL201410539423.0.

[0022] 16. Shi Jiangong, Zhang Jianjun, Yue Zhenggang, et al. N 6 -substitutedadenosine derivatives and N 6 -supposed adenine derivatives and uses thereof[p].US10174033 B2.

[0023] 17. Shi Jiangong, Zhang Jianjun, Zheng Wensheng, et al. A tablet for treating insomnia and its preparation method [p]. 2023112793499. Summary of the Invention

[0024] The technical problem solved by the present invention is to provide the crystal form of compound YZG-331 as shown in the following formula and its preparation method.

[0025]

[0026] To solve the technical problem of the present invention, the present invention provides crystal form A of compound YZG-331.

[0027] Using Cu-Kα radiation, the X-ray powder diffraction of crystal form A exhibits characteristic peaks at diffraction angles 2θ of 17.4°±0.2°, 14.1°±0.2°, and 8.4°±0.2°.

[0028] Furthermore, the X-ray powder diffraction of the crystal form A has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 18.2°±0.2°, 12.5°±0.2°, and 13.1°±0.2°.

[0029] Furthermore, the X-ray powder diffraction of the crystal form A has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 23.7°±0.2°, 24.4°±0.2°, and 6.6°±0.2°.

[0030] In a preferred embodiment, the X-ray powder diffraction of crystal form A exhibits characteristic peaks at diffraction angles 2θ of 17.4°±0.2°, 14.1°±0.2°, 8.4°±0.2°, 18.2°±0.2°, 12.5°±0.2°, 13.1°±0.2°, 23.7°±0.2°, 24.4°±0.2°, and 6.6°±0.2°.

[0031] In a further preferred embodiment, the X-ray powder diffraction pattern of crystal form A has characteristic peaks, d-values, and relative intensities at diffraction angle 2θ as follows:

[0032]

[0033]

[0034] The crystal form A provided by this invention is characterized in that its X-ray powder diffraction pattern is substantially as shown in the attached figure. Figure 1 As shown.

[0035] When performing differential scanning calorimetry (DSC), crystal form A showed an endothermic signal when heated to around 66°C and began to melt when heated to around 102°C. Its DSC data is shown in the attached figure. Figure 2 As shown.

[0036] When thermogravimetric analysis was performed, crystal form A exhibited a mass loss gradient of approximately 2.3% when heated to 100°C. Its TGA is shown in the attached figure. Figure 3 As shown.

[0037] According to the purpose of this invention, the present invention provides crystal form B of compound YZG-331.

[0038] Using Cu-Kα radiation, the X-ray powder diffraction of the crystal form B exhibits characteristic peaks at diffraction angles 2θ of 6.1°±0.2°, 12.2°±0.2°, and 13.7°±0.2°.

[0039] Furthermore, the X-ray powder diffraction of the crystal form B has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 13.3°±0.2°, 17.6°±0.2°, and 6.6°±0.2°.

[0040] Furthermore, the X-ray powder diffraction of the crystal form B has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 12.6°±0.2°, 23.9°±0.2°, and 8.0°±0.2°.

[0041] In a preferred embodiment, the X-ray powder diffraction of crystal form B exhibits characteristic peaks at diffraction angles 2θ of 6.1°±0.2°, 12.2°±0.2°, 13.7°±0.2°, 13.3°±0.2°, 17.6°±0.2°, 6.6°±0.2°, 12.6°±0.2°, 23.9°±0.2°, and 8.0°±0.2°.

[0042] In a further preferred embodiment, the X-ray powder diffraction pattern of crystal form B has characteristic peaks, d-values, and relative intensities at diffraction angle 2θ as follows:

[0043] Diffraction angle 2θ d value Relative strength % 6.1 14.5 100.0 6.6 13.4 56.5 8.0 11.0 27.4 12.2 7.3 62.2 12.6 7.0 56.3 13.3 6.7 57.1 13.7 6.5 59.2 17.6 5.1 57.0 23.9 3.7 28.0

[0044] The crystal form B provided by this invention is characterized in that its X-ray powder diffraction pattern is substantially as shown in the attached figure. Figure 4 As shown.

[0045] When performing differential scanning calorimetry (DSC), crystal form B showed an endothermic signal when heated to around 43°C and began to melt when heated to around 99°C. Its DSC data is shown in the attached figure. Figure 5 As shown.

[0046] When thermogravimetric analysis was performed, crystal form B exhibited a mass loss gradient of approximately 6.8% when heated to 120°C. Its TGA is shown in the attached figure. Figure 6 As shown.

[0047] According to the purpose of this invention, the present invention provides crystal form A1 of compound YZG-331.

[0048] Using Cu-Kα radiation, the X-ray powder diffraction of the crystal form Al exhibits characteristic peaks at diffraction angles 2θ of 11.6°±0.2°, 14.4°±0.2°, and 6.7°±0.2°.

[0049] Furthermore, the X-ray powder diffraction of the crystal form A1 has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 18.5°±0.2°, 20.8°±0.2°, and 12.8°±0.2°.

[0050] Furthermore, the X-ray powder diffraction of the crystal form A1 has characteristic peaks at one, two, or three locations within the diffraction angles 2θ of 8.6°±0.2°, 13.4°±0.2°, and 15.8°±0.2°.

[0051] In a preferred embodiment, the X-ray powder diffraction of the crystal form A1 exhibits characteristic peaks at diffraction angles 2θ of 11.6°±0.2°, 14.4°±0.2°, 6.7°±0.2°, 18.5°±0.2°, 20.8°±0.2°, 12.8°±0.2°, 8.6°±0.2°, 13.4°±0.2°, and 15.8°±0.2°.

[0052] In a further preferred embodiment, the X-ray powder diffraction pattern of the crystal form A1 has characteristic peaks, d-values, and relative intensities at diffraction angles 2θ as follows:

[0053] Diffraction angle 2θ d value Relative strength % 6.7 13.2 83.5 8.6 10.3 30.8 11.6 7.6 100.0 12.8 6.9 55.1 13.4 6.6 19.3 14.4 6.1 90.4 15.8 5.6 17.2 18.5 4.8 69.0 20.8 4.3 59.3

[0054] The crystal form A1 provided by this invention is characterized in that its X-ray powder diffraction pattern is substantially as shown in the attached figure. Figure 7 As shown.

[0055] When performing differential scanning calorimetry (DSC), crystal form A1 showed an endothermic signal when heated to around 51°C and began to melt when heated to around 97°C. Its DSC data is shown in the attached figure. Figure 8 As shown.

[0056] When thermogravimetric analysis was performed, crystal form Al exhibited a mass loss gradient of approximately 6.7% when heated to 120°C. Its TGA is shown in the attached figure. Figure 9 As shown.

[0057] Beneficial technical effects

[0058] In the process of researching the sedative-hypnotic candidate drug YZG-331, the inventors of this invention discovered three crystal forms of YZG-331 through various experimental techniques and verified the crystal form preparation methods, laying the foundation for the subsequent development of YZG-331. These are novel crystal forms of the sedative-hypnotic candidate drug YZG-331. Attached Figure Description

[0059] Figure 1 X-ray powder diffraction pattern of crystal form A

[0060] Figure 2 Differential scanning calorimetry (DSC) plot of crystal form A

[0061] Figure 3 Thermogravimetric analysis (TGA) diagram of crystal form A

[0062] Figure 4 X-ray powder diffraction pattern of crystal form B

[0063] Figure 5 Differential scanning calorimetry (DSC) plot of crystal form B

[0064] Figure 6 Thermogravimetric analysis (TGA) diagram of crystal form B

[0065] Figure 7 X-ray powder diffraction pattern of crystal form A1

[0066] Figure 8 Differential scanning calorimetry (DSC) plot of crystal form A1

[0067] Figure 9 Thermogravimetric analysis (TGA) diagram of crystal form A1

[0068] Figure 10 Dynamic water adsorption-desorption (DVS) curves of crystal form A

[0069] Figure 11 Dynamic water adsorption-desorption (DVS) curves of amorphous YZG-331 Detailed Implementation

[0070] The present invention will be further illustrated by specific embodiments below, but these are not intended to limit the scope of protection of the present invention. Those skilled in the art can make improvements to the preparation method and the instruments used within the scope of the claims, and these improvements should also be considered within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

[0071] The abbreviations used in this invention are explained as follows:

[0072] XRPD: X-ray powder diffraction

[0073] DSC: Differential Scanning Calorimetry

[0074] TGA: Thermogravimetric Analysis

[0075] DVS: Dynamic Moisture Adsorption

[0076] The X-ray powder diffraction pattern described in this invention is in PANalytical Empyrean or X'Pert. 3 Data was collected using an X-ray powder diffractometer. The parameters of the X-ray powder diffraction method described in this invention are as follows:

[0077] X-ray source: Cu, Kα

[0078] 1.54060; 1.54439

[0079] Kα2 / Kα1 intensity ratio: 0.50

[0080] Voltage: 45 kV

[0081] Current: 40 milliamperes (mA)

[0082] Diverging slit: 1 / 8 degree

[0083] Scanning mode: Continuous scan

[0084] Scan range: from 3.0 to 40.0 degrees

[0085] Scan step size: 0.0263 degrees

[0086] Scanning time per step: 46.67 degrees

[0087] Scan time: Approximately 5 minutes

[0088] The differential scanning calorimetry (DSC) images described in this invention were acquired on a TA Discovery 2500. The parameters of the differential scanning calorimetry (DSC) method described in this invention are as follows:

[0089] Method: Linear heating

[0090] Sample tray: Aluminum tray, open

[0091] Temperature range: room temperature to 350℃

[0092] Scan rate: 10℃ / minute

[0093] Protective gas: N2

[0094] The thermogravimetric analysis (TGA) charts described in this invention were acquired using a TA Discovery 5500. The method parameters for the thermogravimetric analysis (TGA) described in this invention are as follows:

[0095] Method: Linear heating

[0096] Sample tray: Aluminum tray, pressure cap

[0097] Temperature range: room temperature to 300℃

[0098] Scan rate: 10℃ / minute

[0099] Protective gas: N2

[0100] The dynamic moisture adsorption map described in this invention is acquired using the DVS Intrinsic module of an SMS system. The parameters of the dynamic moisture adsorption method described in this invention are as follows:

[0101] Temperature: 25℃

[0102] Sample size: 20 to 40 mg

[0103] Protective gas and flow rate: N2, 200 ml / min

[0104] Maximum mass change at equilibrium, dm / dt: 0.002% / minute

[0105] Minimum dm / dt equilibration time: 10 minutes

[0106] Maximum balancing time: 180 minutes

[0107] Humidity range: Room humidity up to 95%RH to 0%RH to 95%RH

[0108] Humidity variation gradient: 10%RH (between 0%RH and 90%RH), 5%RH (between 90%RH and 95%RH)

[0109] In this invention, the high-performance liquid chromatography (HPLC) purity data were obtained from an Agilent 1260, and the detector used was a diode array detector (DAD). The HPLC method parameters for testing purity described in this invention are as follows:

[0110] 1. Chromatographic column: Zorbax Eclipse XDB-C18, 150×4.6mm, 5μm

[0111] 2. Mobile phase: A: 0.1% aqueous acetic acid solution

[0112] B: Methanol

[0113] The elution gradient is as follows:

[0114]

[0115]

[0116] 3. Flow rate: 0.8 mL / min

[0117] 4. Injection volume: 2 μL or 10 μL

[0118] 5. Detection wavelength: 269nm

[0119] 6. Column temperature: 40℃

[0120] 7. Injector temperature: room temperature

[0121] 8. Diluent: 0.1% methanol / water (85:15, volume ratio) solution

[0122] Unless otherwise specified, all the following examples are performed at room temperature.

[0123] Example 1: Preparation of crystal form A

[0124] Using 6-chloroadenosine and (S)-α-ethylbenzylamine as starting materials, sodium carbonate as an acid scavenger, and reflux in anhydrous ethanol, the compound YZG-331 can be obtained after one-step reaction and recrystallization purification.

[0125] Upon testing, the solid obtained in this embodiment was found to be crystal form A. The X-ray powder diffraction data of the obtained solid are shown in Table 1. DSC and TGA tests were performed on the above-mentioned crystal form A sample, and the data are as follows: Figure 2 and Figure 3 As shown.

[0126] Table 1

[0127] Diffraction angle 2θ d value Relative strength % 6.6 13.5 23.7 8.4 10.5 84.3 11.4 7.8 14.8 12.5 7.1 59.8 13.1 6.7 57.4 14.1 6.3 90.5 15.5 5.7 19.2 16.8 5.3 3.3 17.4 5.1 100.0 18.2 4.9 65.0 19.6 4.5 4.2 20.4 4.3 63.6 20.6 4.3 22.4 22.2 4.0 8.2 23.1 3.8 1.9 23.7 3.7 37.8 24.4 3.7 26.0 26.1 3.4 7.1 26.4 3.4 10.1 28.6 3.1 3.7 30.8 2.9 5.2 32.1 2.8 4.0

[0128] Example 2: Preparation of crystal form B

[0129] Weigh 21.1 mg of compound YZG-331 solid into a 2 mL glass vial, add 0.5 mL of methyl tert-butyl ether to obtain a suspension. Stir the suspension magnetically at 5 °C for about 4 days.

[0130] Upon testing, the solid obtained in this embodiment was found to be crystal form B. The X-ray powder diffraction data of the obtained solid are shown in Table 2. DSC and TGA tests were performed on the above crystal form B sample, and the data are as follows: Figure 5 and Figure 6 As shown.

[0131] Table 2

[0132] Diffraction angle 2θ d value Relative strength % 5.0 17.7 15.5 6.1 14.5 100.0 6.6 13.4 56.5 7.3 12.1 5.7 8.0 11.0 27.4 8.5 10.4 12.6 9.2 9.6 8.1 11.0 8.0 13.6 11.4 7.7 22.2 12.2 7.3 62.2 12.6 7.0 56.3 13.3 6.7 57.1 13.7 6.5 59.2 14.2 6.2 21.7 14.6 6.1 16.5 15.1 5.9 14.2 15.4 5.7 19.1 15.7 5.6 24.6 16.7 5.3 16.7 17.1 5.2 19.8 17.6 5.1 57.0 18.0 4.9 16.5 18.3 4.8 60.1 19.2 4.6 10.1 19.9 4.5 8.6 20.5 4.3 25.4 21.3 4.2 11.5 23.9 3.7 28.0

[0133] Example 3: Preparation of crystal form A1

[0134] Weigh 20.0 mg of compound YZG-331 into a 3 mL glass vial. Transfer 4 mL of ethyl acetate into a 20 mL glass vial. Place the 3 mL vial containing the solid sample open into the 20 mL vial, tighten the cap of the 20 mL vial, and leave it at room temperature for 10 days. Transfer the solid to room temperature and humidity for 1 day.

[0135] Upon testing, the solid obtained in this embodiment was found to be crystalline form A1. The X-ray powder diffraction data of the obtained solid are shown in Table 3. DSC and TGA tests were performed on the above-mentioned crystalline form A1 sample, and the data are as follows: Figure 8 and Figure 9 As shown.

[0136] Table 3

[0137] Diffraction angle 2θ d value Relative strength % 6.7 13.2 83.5 8.6 10.3 30.8 11.6 7.6 100.0 12.8 6.9 55.1 13.4 6.6 19.3 14.4 6.1 90.4 15.8 5.6 17.2 17.2 5.2 7.6 17.4 5.1 6.7 17.7 5.0 43.5 18.5 4.8 69.0 19.9 4.5 2.1 20.8 4.3 59.3 22.5 3.9 12.8 22.9 3.9 5.4 23.3 3.8 2.2 24.0 3.7 24.0 24.2 3.7 36.4 24.8 3.6 14.3 26.6 3.4 10.1 26.9 3.3 6.4 27.8 3.2 3.4 29.2 3.1 5.0 29.6 3.0 2.5 31.4 2.8 5.4 32.5 2.8 3.3 33.9 2.6 2.4 34.3 2.6 2.9 34.9 2.6 2.6

[0138] Examples 4-5: Preparation of crystal form A (antisolvent addition method)

[0139] Weigh approximately 20 mg of compound YZG-331 solid into a 20 mL glass vial, add the corresponding volume of positive solvent, and filter the sample solution into a new 20 mL glass vial using a 0.45 μm pore size polytetrafluoroethylene filter membrane. Then, magnetically stir the resulting clear solution (at approximately 1000 rpm), and add the corresponding antisolvent dropwise. If solid precipitates, centrifuge to separate the solid. If an oil is formed, circulate the suspension at 5–50 °C with stirring (program: 50 °C for 2 hours, decrease to 5 °C at a rate of 0.1 °C / min, maintain at 5 °C for 2 hours, then increase to 50 °C over 0.5 hours. Repeat this cycle 3 times, decreasing to 5 °C at a rate of 0.1 °C / min and maintaining at 5 °C with stirring).

[0140] The detailed experimental conditions involved in the above embodiments are shown in Table 4. Testing revealed that the solids obtained in the above embodiments were all of crystal form A. Taking Example 5 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 5.

[0141] Table 4

[0142]

[0143] Table 5

[0144] Diffraction angle 2θ d value Relative strength % 6.6 13.4 100.0 8.5 10.4 24.3 11.4 7.8 75.2 12.6 7.0 61.6 13.2 6.7 27.7 14.2 6.2 79.0 15.7 5.7 12.8 17.0 5.2 6.3 17.4 5.1 74.1 18.2 4.9 75.3 20.5 4.3 76.9 22.5 3.9 9.7 23.5 3.8 10.5 23.8 3.7 54.9 24.4 3.6 28.4 26.2 3.4 7.2 26.5 3.4 10.9 27.0 3.3 2.7 27.8 3.2 2.0 28.9 3.1 5.6 29.4 3.0 2.6 30.9 2.9 8.4

[0145] Examples 6-8: Preparation of crystal form A (gas-solid diffusion method)

[0146] Weigh approximately 20 mg of the solid compound YZG-331 into a 3 mL glass vial. Transfer 4 mL of solvent into a 20 mL glass vial. Place the 3 mL vial containing the solid sample open into the 20 mL vial, tighten the cap of the 20 mL vial, and store at room temperature.

[0147] The detailed experimental conditions involved in the above embodiments are shown in Table 6. Testing revealed that the solids obtained in the above embodiments were all of crystal form A. Taking Example 7 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 7.

[0148] Table 6

[0149]

[0150] Table 7

[0151] Diffraction angle 2θ d value Relative strength % 6.6 13.4 89.9 8.5 10.4 61.2 11.4 7.8 82.3 12.6 7.0 93.3 13.2 6.7 21.8 14.2 6.2 100.0 15.7 5.6 24.7 17.0 5.2 14.2 17.5 5.1 57.8 18.3 4.8 80.9 20.6 4.3 71.3 22.6 3.9 22.2 23.6 3.8 14.1 23.8 3.7 56.1 24.5 3.6 22.9 26.3 3.4 8.1 27.8 3.2 8.7 29.0 3.1 7.0 29.5 3.0 5.0 31.0 2.9 5.3 32.4 2.8 5.6 33.1 2.7 2.4 33.8 2.6 4.9 34.7 2.6 7.7

[0152] Example 9: Preparation of crystal form A (80°C suspension stirring method)

[0153] Weigh 21.7 mg of compound YZG-331 solid into a 2 mL glass vial, add 0.5 mL of m-xylene solvent to obtain a suspension. Stir the suspension magnetically at 80 °C for about 4 days.

[0154] Upon testing, the solid obtained in the above embodiment was crystal form A, and the X-ray powder diffraction data of the obtained solid are shown in Table 8.

[0155] Table 8

[0156]

[0157]

[0158] Example 10: Preparation of Crystal Form A (Solvothermal Method)

[0159] Weigh 20.8 mg of compound YZG-331 solid into a 3 mL glass vial, add 1 mL of methyl tert-butyl ether, and place the 3 mL vial into a hydrothermal reactor with a Teflon liner. Place the reactor in a biochemical incubator for temperature regulation (temperature program: increase from 25°C to 80°C within 6 hours, hold at 80°C for 12 hours, and decrease to 5°C at a rate of 0.05°C / min), and then hold at 5°C.

[0160] Upon testing, the solid obtained in the above embodiment was found to be crystal form A. The X-ray powder diffraction data of the obtained solid are shown in Table 9.

[0161] Table 9

[0162]

[0163]

[0164] Examples 11-19: Preparation of Crystal Form A1 (Antisolvent Addition Method)

[0165] Weigh approximately 20 mg of compound YZG-331 solid into a 20 mL glass vial, add an appropriate volume of positive solvent, and filter the sample solution into a new 20 mL glass vial using a 0.45 μm pore size polytetrafluoroethylene filter membrane. Then, magnetically stir the resulting clear solution (at approximately 1000 rpm) and add the appropriate antisolvent dropwise. If solid precipitates, centrifuge to separate the solid. If the solution remains clear, suspend and stir at 5 °C.

[0166] The detailed experimental conditions involved in the above embodiments are shown in Table 10. Upon testing, the solids obtained in the above embodiments were all of crystal form Al. Taking Example 15 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 11.

[0167] Table 10

[0168]

[0169] Table 11

[0170]

[0171]

[0172] Examples 20-26: Preparation of crystal form A1 (gas-solid diffusion method)

[0173] Weigh approximately 20 mg of the solid compound YZG-331 into a 3 mL glass vial. Transfer 4 mL of solvent into a 20 mL glass vial. Place the 3 mL vial containing the solid sample open into the 20 mL vial, tighten the cap of the 20 mL vial, and store at room temperature.

[0174] The detailed experimental conditions involved in the above embodiments are shown in Table 12. Testing revealed that the solids obtained in the above embodiments were all of crystal form Al. Taking Example 23 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 13.

[0175] Table 12

[0176]

[0177]

[0178] Table 13

[0179] Diffraction angle 2θ d value Relative strength % 6.7 13.2 95.1 8.6 10.3 34.2 11.5 7.7 90.8 12.0 7.5 1.0 12.7 6.9 76.9 13.3 6.6 25.6 14.4 6.2 100.0 15.8 5.6 29.9 17.1 5.2 12.5 17.4 5.1 12.9 17.7 5.0 56.5 18.5 4.8 92.6 19.8 4.5 7.2 20.8 4.3 72.0 22.5 3.9 26.5 22.8 3.9 6.6 24.0 3.7 34.9 24.2 3.7 39.9 24.7 3.6 24.3 26.5 3.4 13.6 26.9 3.3 9.5 27.8 3.2 7.9 29.1 3.1 5.2 29.5 3.0 4.8 31.3 2.9 7.3 32.6 2.7 6.9 33.7 2.7 4.9 35.0 2.6 8.4

[0180] Examples 27-30: Preparation of crystal form A1 (slow evaporation method)

[0181] Weigh approximately 20 mg of compound YZG-331 solid into a 3 mL glass vial, add the corresponding volume of solvent, and filter the sample solution into a new 3 mL glass vial using a 0.45 μm pore size polytetrafluoroethylene filter membrane. Seal the 3 mL glass vial with sealing film, make 4 small holes in the sealing film, and leave it at room temperature to evaporate until solid precipitates.

[0182] The detailed experimental conditions involved in the above embodiments are shown in Table 14. Upon testing, the solids obtained in the above embodiments were all of crystal form Al. Taking Example 28 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 15.

[0183] Table 14

[0184]

[0185]

[0186] Table 15

[0187] Diffraction angle 2θ d value Relative strength % 6.6 13.3 100.0 8.5 10.4 1.0 11.5 7.7 31.9 12.7 7.0 3.5 13.3 6.7 60.2 14.3 6.2 7.4 17.1 5.2 0.7 17.6 5.0 29.0 18.4 4.8 12.3 20.7 4.3 16.7 23.1 3.8 1.6 23.8 3.7 3.8 24.1 3.7 26.0 24.7 3.6 7.9 26.4 3.4 1.7 26.7 3.3 26.6 27.2 3.3 0.4 29.7 3.0 0.7 30.8 2.9 0.7 31.2 2.9 3.7 33.6 2.7 8.2 34.0 2.6 2.4 37.5 2.4 1.6 39.2 2.3 1.2

[0188] Examples 31-35: Preparation of Crystal Form A1 (Slow Cooling Method)

[0189] Approximately 20 mg of compound YZG-331 solid was weighed and placed in a 2 mL glass vial. The corresponding solvent was added to obtain a suspension. The suspension was magnetically stirred at 50 °C (approximately 1000 rpm) for about two hours. The sample solution was then filtered hot through a 0.45 μm pore size polytetrafluoroethylene (PTFE) filter into a new 2 mL glass vial. After sealing, the vial was cooled from 50 °C to 5 °C at a rate of 0.1 °C per minute.

[0190] The detailed experimental conditions involved in the above embodiments are shown in Table 16. Testing revealed that the solids obtained in the above embodiments were all of crystal form Al. Taking Example 32 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 17.

[0191] Table 16

[0192]

[0193] Table 17

[0194]

[0195]

[0196] Examples 36-41: Preparation of Crystal Form A1 (Gas-Liquid Permeation Method)

[0197] Weigh approximately 20 mg of compound YZG-331 solid into a 3 mL glass vial. Dissolve the solid in solvent. Filter the sample solution through a 0.45 μm pore size polytetrafluoroethylene (PTFE) membrane into a new 3 mL glass vial. Place the vial open into a 20 mL glass vial pre-filled with 4 mL of the corresponding antisolvent. Seal the vial and allow it to stand at room temperature.

[0198] The detailed experimental conditions involved in the above embodiments are shown in Table 18. Upon testing, the solids obtained in the above embodiments were all of crystal form Al. Taking Example 41 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 19.

[0199] Table 18

[0200]

[0201] Table 19

[0202]

[0203]

[0204] Examples 42-50: Preparation of Crystal Form A1 (Room Temperature Suspension Stirring Method)

[0205] Weigh approximately 20 mg of the solid into a 2 mL glass vial, and add an appropriate volume of solvent to obtain a suspension. Stir the suspension magnetically at room temperature for approximately 4 days.

[0206] The detailed experimental conditions involved in the above embodiments are shown in Table 20. Upon testing, the solids obtained in the above embodiments were all of crystal form Al. Taking Example 47 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 21.

[0207] Table 20

[0208]

[0209]

[0210] Table 21

[0211] Diffraction angle 2θ d value Relative strength % 6.7 13.2 39.9 8.6 10.3 26.9 10.8 8.2 0.0 11.5 7.7 61.0 12.7 7.0 59.9 13.3 6.6 13.9 14.4 6.2 88.4 15.8 5.6 21.8 17.1 5.2 12.3 17.6 5.0 53.9 18.4 4.8 94.9 19.9 4.5 3.8 20.7 4.3 100.0 20.9 4.2 12.5 22.6 3.9 20.4 23.1 3.8 3.4 23.7 3.7 16.2 24.0 3.7 60.3 24.7 3.6 34.5 25.4 3.5 3.9 26.4 3.4 20.0 26.7 3.3 9.4 27.9 3.2 11.2 29.1 3.1 10.8 29.5 3.0 6.7 29.7 3.0 2.1 30.2 3.0 1.7 31.1 2.9 13.1 31.7 2.8 0.0 31.8 2.8 2.8 32.5 2.8 11.1 33.5 2.7 3.8 34.0 2.6 5.3 34.8 2.6 11.8 35.3 2.5 2.9 36.0 2.5 2.3 36.5 2.5 2.1 37.5 2.4 2.6 39.1 2.3 2.7

[0212] Examples 51-59: Preparation of crystal form A1 (80°C suspension stirring method)

[0213] Approximately 20 mg of the compound YZG-331 solid was weighed and placed in a 2 mL glass vial. An appropriate volume of solvent was added to obtain a suspension. The suspension was magnetically stirred at 80 °C for approximately 4 days.

[0214] The detailed experimental conditions involved in the above embodiments are shown in Table 22. Upon testing, the solids obtained in the above embodiments were all of crystal form Al. Taking Example 58 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 23.

[0215] Table 22

[0216]

[0217] Table 23

[0218]

[0219]

[0220] Examples 60-61: Preparation of crystal form A1 (solvothermal method)

[0221] Approximately 20 mg of compound YZG-331 solid was weighed and placed in a 3 mL glass vial. An appropriate volume of solvent was added, and the vial was placed in a Teflon-lined hydrothermal reactor. The reactor was then placed in a biochemical incubator for temperature regulation (temperature program: increasing from 25°C to 80°C over 6 hours, holding at 80°C for 12 hours, and decreasing to 5°C at a rate of 0.05°C / min), followed by holding at 5°C. Detailed experimental conditions in the above examples are shown in Table 24. The solids obtained in the above examples were all of crystal form Al. Taking Example 61 as an example, the X-ray powder diffraction data of the obtained solid are shown in Table 25.

[0222] Table 24

[0223]

[0224] Table 25

[0225]

[0226]

[0227] Example 62: Hygroscopicity

[0228] To evaluate the hygroscopicity of crystal form A, the dynamic water adsorption-desorption (DVS) curves of crystal form A and the amorphous sample prepared in Example 1 of this invention were tested respectively. The experimental results are shown in Table 26. The DVS of crystal form A is attached. Figure 10 As shown. The amorphous DVS is attached. Figure 11 As shown.

[0229] Table 26

[0230]

[0231] According to the definition of hygroscopicity of drugs in the 2020 edition of the Chinese Pharmacopoeia, both crystal form A and crystal form B described in this invention are hygroscopic with little or no hygroscopicity. They can maintain their properties under high humidity conditions and can remain stable without specific humidity control conditions during production, storage and use, thus well meeting the requirements of drug production and use.

[0232] Description of hygroscopic characteristics and definition of hygroscopic weight gain (Guidelines for Hygroscopicity Testing of Drugs, Chinese Pharmacopoeia 2020 Edition):

[0233] Deliquescence: Absorbs sufficient water to form a liquid.

[0234] Extremely hygroscopic: Moisture absorption increases weight by at least 15%.

[0235] It has hygroscopic properties: the weight gain due to moisture absorption is less than 15% but not less than 2%.

[0236] Slightly hygroscopic: Moisture absorption increases weight by less than 2% but not less than 0.2%.

[0237] None or almost none hygroscopicity: moisture-induced weight gain less than 0.2%.

[0238] Example 63: Stability Comparison Study

[0239] Weigh approximately 20 mg of crystal form A (initial purity 99.49 area%) and amorphous sample (initial purity 99.44 area%) obtained in Preparation Example 1 of this invention. Place them closed in an oven at 80°C. After one day, take samples to measure XRPD and HPLC purity. Place them open in a stabilization chamber at 25°C / 60%RH and 40°C / 75%RH. After one week, take samples to measure XRPD and HPLC purity. Place them closed in a light stabilization chamber with a total illuminance of 1,200,000 Lux·hrs for white light and 200 W·hrs / m² for ultraviolet light. 2 The sample was taken at 25℃ after one week to determine its XRPD and HPLC purity. The experimental results are shown in Table 27. Crystal form A transformed into amorphous under 40℃ / 75%RH conditions, and the crystal form did not change significantly under other conditions. The purity of crystal form A did not change significantly under all conditions. The crystal form and purity of amorphous crystals did not change significantly under all conditions.

[0240] Table 27

[0241]

[0242] * A small transparent glass bottle was wrapped in aluminum foil and placed in a light-stabilized chamber under the same lighting conditions as a control experiment.

[0243] Example 64: Solubility of the Crystal Form

[0244] Approximately 20 mg of the crystalline form A and amorphous sample obtained in Preparation Example 1 of this invention were weighed and prepared into suspensions using 2 mL of SGF (simulated gastric juice), FaSSIF (artificial intestinal juice under fasting conditions), and FeSSIF (artificial intestinal juice under feeding conditions), respectively. After equilibration at 37°C for 24 hours, the suspensions were filtered to obtain clear solutions. The sample content in the saturated solution was determined by HPLC. The experimental results are shown in Table 28.

[0245] Weigh approximately 20 to 40 mg of the crystalline form A and amorphous sample obtained in Preparation Example 1 of this invention, and equilibrate them with 2 mL of pure water, pH 1.0 buffer, pH 4.5 buffer, and pH 6.8 buffer at room temperature for 24 hours, respectively, and then filter to obtain a clear solution. The sample content in the saturated solution was determined by HPLC. The experimental results are shown in Table 29.

[0246] The experimental results showed that in FeSSIF and pH 6.8 buffer, the solubility of crystalline form A was higher than that of amorphous form; in FaSSIF, the solubility of amorphous form was higher than that of crystalline form A; and in SGF, H2O, pH 1.0 and pH 4.5 buffers, there was no significant difference in solubility between crystalline form A and amorphous form.

[0247] Table 28

[0248]

[0249] Table 29

[0250]

[0251] Example 65: Bulk density and tap density of crystal form

[0252] Weigh approximately 500 mg of both the crystalline form A and the amorphous sample obtained in Preparation Example 1 of this invention, and add them separately to a 5 mL graduated cylinder, recording the volume at this point. Tap the graduated cylinder gently on the table 200 times, recording the volume again. Perform three parallel tests. The experimental results are shown in Table 30.

[0253] Table 30

[0254]

[0255] Example 66: Angle of Repose of Crystal Form

[0256] Take appropriate amounts of both crystalline form A and amorphous sample obtained in Preparation Example 1 of this invention. Fix a funnel on an iron stand, placing it perpendicular to the tabletop. Add the crystalline form A and amorphous sample to the funnel respectively, allowing them to fall freely and form a cone. Measure the height and diameter of the cone to calculate the angle of repose. Perform three parallel tests. The experimental results are shown in Table 31. The results show that there is no significant difference in the flowability of crystalline form A and the amorphous sample.

[0257] Table 31

[0258]

[0259] Example 67: Comparative Study of Particle Size Distribution

[0260] Weigh appropriate amounts of crystalline form A and amorphous sample obtained in Preparation Example 1 of this invention, add ISOPAR G dispersion medium, mix the samples thoroughly, and add 10-30 mg of sample to the injection system for particle size distribution testing. The experimental results are shown in Table 32. Crystalline form A of this invention exhibits a unimodal distribution, while the amorphous form exhibits a bimodal distribution with some overlap.

[0261] Table 32

[0262] Crystal form Volume average particle size (micrometers) D10 (micrometers) D50 (micrometers) D90 (micrometers) Crystal form A 57.79 17.95 39.27 122.3 amorphous 154.8 40.35 132.5 298.2

[0263] Example 68: Contact Angle of Crystal Form

[0264] Weigh appropriate amounts of both the crystalline form A and the amorphous sample obtained in Preparation Example 1 of this invention, and test their contact angle with water. Two parallel tests were performed. The test results are shown in Table 33.

[0265] Table 33

[0266] Crystal form Test 1 Test 2 Crystal form A 81.5°～84.6° 79.2°～84.6° amorphous 56.1°～56.8° 54.5°～56.2°

[0267] Example 69: Mechanical stability of crystal form

[0268] Appropriate amounts of crystal form A and the amorphous sample obtained in Preparation Example 1 of this invention were placed in an agate mortar and manually ground for about 3 minutes before the crystal form was tested. The test results showed that crystal form A did not undergo a crystal form transformation after grinding, but the crystallinity decreased slightly, while the amorphous sample remained amorphous.

[0269] Appropriate amounts of crystal form A and the amorphous sample obtained in Preparation Example 1 of this invention were added to a tablet press and tableted at 350 MPa. The crystal form was then tested. The test results showed that crystal form A did not undergo a crystal form transformation during tableting, but its crystallinity decreased slightly, while the amorphous sample remained amorphous.

[0270] The above embodiments are only for illustrating the technical concept and features of the present invention, and are intended to enable those skilled in the art to understand the content of the present invention and implement it accordingly. They should not be construed as limiting the scope of protection of the present invention. All equivalent changes or modifications made in accordance with the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. Crystal form A of compound YZG-331 shown in formula (I), characterized in that, Using Cu-Kα radiation, the X-ray powder diffraction of crystal form A exhibits characteristic peaks, d-values, and relative intensities at the following positions with a 2θ value:

2. The method for preparing crystal form A according to claim 1, characterized in that, The compound of formula (I) was dissolved in at least one of the positive solvents methanol and isopropanol. After filtration, the antisolvents toluene and cyclohexane were added dropwise. The system was stirred and the temperature was set to rise and fall between 5°C and 50°C for 3 cycles to obtain crystal form A.

3. The method for preparing crystal form A according to claim 1, characterized in that, When the compound of formula (I) is placed in an atmosphere of at least one of methyl tert-butyl ether and toluene, the solid undergoes gas-solid diffusion with the volatile solvent to obtain crystal form A.

4. The method for preparing crystal form A according to claim 1, characterized in that, Compound of formula (I) was added to methyl tert-butyl ether, sealed in a hydrothermal reactor, and the reactor was placed under high temperature and high pressure conditions for reaction. After the reaction was carried out, the temperature was slowly lowered to 5°C to precipitate a solid, yielding crystal form A.

5. The method for preparing crystal form A according to claim 1, characterized in that, An excess of compound (I) was added to the methyl tert-butyl ether system, and the suspension was stirred at 80°C to obtain crystal form A.

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