Salt form of novel cell cycle regulatory kinase inhibitor, crystal form thereof and preparation method therefor

Abstract

The present invention relates to a salt form of a novel cell cycle regulatory kinase inhibitor, a crystal form thereof, and a preparation method therefor. A succinate salt of a compound (6‑((4‑(2‑(diethylamino)benzo[d]thiazol‑6‑yl)‑5‑fluoropyrimidin‑2‑yl)amino)pyridin‑3‑yl)(4‑isopropylpiperazin‑1‑yl)methanone and a crystal form thereof are obtained by screening, wherein two crystal forms II and III of the succinate salt have good stability and high solubility, and the preparation method therefor is simple and suitable for industrial production.

Description

A novel cell cycle-regulated kinase inhibitor salt form, its crystal form, and preparation method Technical Field

[0001] This invention belongs to the fields of medicinal chemistry and crystals, and specifically relates to a novel cell cycle regulation kinase inhibitor salt form, its crystal form, and preparation method. Background Technology

[0002] Cancer is a major global health challenge, severely impacting human health and lives. According to the 2024 Global and China Cancer Reports, the global incidence and mortality rates of cancer are rising annually. Abnormal proliferation caused by cell cycle imbalance is a fundamental characteristic of tumor cells and is closely related to the occurrence, development, recurrence, drug resistance, and metastasis of tumors. Intervening in key nodes of the tumor cell cycle can effectively inhibit tumor cell growth and induce apoptosis, becoming an important mechanism of action for many anti-tumor drugs. Therefore, developing inhibitors targeting key regulatory proteins of the cell cycle has become one of the core directions in anti-tumor drug development.

[0003] Both CDK4 / 6 and DYRK2 are key targets for cell cycle regulation. CDK4 / 6 are serine / threonine kinases that bind to cyclin D, arresting the transition of tumor cells from G1 to S phase, thus producing anti-proliferative and anti-cancer effects. Currently marketed CDK4 / 6 inhibitors include palbociclib, abeciclib, ribociclib, and dalciribé. These inhibitors inhibit tumor cell proliferation by suppressing CDK4 / 6 activity. They are now widely used as first-line treatments for ER+ / HER2- advanced breast cancer. However, most patients develop resistance after using these inhibitors, which is a current clinical challenge. DYRK2 is a newly discovered pan-cancer target for cell cycle regulation in recent years. Its aberrant expression or altered activity plays an important role in the occurrence and development of various cancers. DYRK2 regulates the cell cycle, inhibiting and slowing the transition from G1 to S phase, while also blocking CDK4 / 6 resistance caused by compensatory pathways. CDK4 / 6 and DYRK2 dual-target inhibitors can act simultaneously on two key targets that regulate the cell cycle, exhibiting a synergistic effect that can enhance the efficacy of treating various tumors and overcome drug resistance.

[0004] The prior art discloses a series of dual-target inhibitor compounds of CDK4 / 6 and DYRK2, among which the representative compound (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone (I-51) has the structure shown below. The existing technology does not disclose the pharmaceutical salt of this compound, nor does it disclose the crystalline form of the pharmaceutical salt. During the development of this compound, the inventors discovered that the free base solubility of this compound is only 0.85 μg / ml, and it also suffers from poor powder properties, difficulties in formulation development processes and quality control. Therefore, the inventors conducted a thorough study of the physicochemical properties of the compound and unexpectedly discovered that its salt form and accompanying crystalline form can solve the aforementioned problems. Thus, the technical solution of this invention is presented. Summary of the Invention

[0005] The purpose of this invention is to provide a novel salt form and crystal form of a cell cycle-regulating kinase inhibitor, and its preparation method. Specifically, the first aspect of this invention is as follows:

[0006] A pharmaceutically acceptable salt of compound I (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl) methyl ketone,

[0007] The salt is selected from: hydrochloride, phosphate, methanesulfonate, hydrobromide, sulfate, p-benzenesulfonate, oxalate, maleate, fumarate, succinate, tartrate, and citrate.

[0008] Preferably, the pharmaceutically acceptable salt of the present invention is (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

[0009] Second aspect of the present invention:

[0010] The crystal form of a pharmaceutically acceptable salt of compound I (6-((4-(2-(diethylamino)benzo[d]thiazo-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl) methyl ketone, preferably the crystal form of compound (6-((4-(2-(diethylamino)benzo[d]thiazo-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl) methyl ketone succinate).

[0011] The compound (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate crystal forms of the present invention include crystal form II and crystal form III: wherein crystal form II has characteristic peaks at 5.3±0.2°, 10.6±0.2°, 15.1±0.2°, 16.1±0.2°, 18.3±0.2°, and 21.1±0.2° when X-ray powder diffraction is performed using Cu-Kα radiation and expressed in 2θ angles.

[0012] The preferred crystal form II, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 5.3±0.2°, 10.6±0.2°, 14.0±0.2°, 15.1±0.2°, 16.1±0.2°, 16.9±0.2°, 18.3±0.2°, 19.5±0.2°, 21.1±0.2°, 23.8±0.2°, 25.5±0.2°, 26.8±0.2°, and 28.2±0.2°.

[0013] More preferably, the crystal form II is obtained using Cu-Kα radiation, with X-ray powder diffraction at angles of 2θ at 5.3±0.2°, 9.2±0.2°, 9.6±0.2°, 10.6±0.2°, 12.0±0.2°, 13.4±0.2°, 14.0±0.2°, 15.1±0.2°, 16.1±0.2°, and 16.9± Characteristic peaks are observed at 0.2°, 18.3±0.2°, 18.5±0.2°, 19.5±0.2°, 21.1±0.2°, 22.7±0.2°, 23.3±0.2°, 23.8±0.2°, 24.5±0.2°, 25.5±0.2°, 26.8±0.2°, 28.2±0.2°, and 32.2±0.2°.

[0014] More preferably, the crystal form II has an X-ray powder diffraction pattern as shown in FIG26;

[0015] Most preferably, the crystal form II is irradiated with Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, is shown in Figure 26 with an error range of ±0.2°.

[0016] The differential scanning calorimetry curve of crystal form II described in this invention has an endothermic peak at 172.8±3℃.

[0017] Preferably, the DSC spectrum of crystal form II is shown in Figure 25.

[0018] The preparation method of crystal form II according to the present invention includes the following steps: adding the free base of compound I and succinate in a certain equivalence ratio to an organic solvent, adjusting to a certain temperature, and stirring until dissolved and clear; cooling and maintaining the temperature for a certain time, then adding an organic solvent containing succinic acid, then adding seed crystal II, maintaining the temperature and stirring, filtering and drying to obtain crystal form II.

[0019] Wherein, the equivalence ratio of the free base of Formula I compound to succinic acid is 1:0.5-2.0, preferably 1:0.8-1.2, and most preferably 1:1;

[0020] The organic solvent is selected from one or two of anisole, ethanol, diethyl ether, dichloromethane, tetrahydrofuran, toluene, ethyl acetate, acetone, methanol, and N,N-dimethylformamide; preferably, the organic solvent is selected from one or two of anisole and ethanol.

[0021] The temperature adjustment is 20-30℃, preferably 30℃;

[0022] The insulation temperature is 20-30℃, preferably 20℃;

[0023] The heat preservation time is 0.5-2 hours, preferably 1 hour.

[0024] Preferably, the preparation method of crystal form II of the present invention comprises the following steps: adding anisole to compound I and succinic acid at an equivalent ratio of 1:1 and stirring, adjusting the temperature to 30°C and stirring until dissolved and clear; cooling to 20°C and maintaining the temperature for 1 hour, adding an ethanol solution of succinic acid; adding an appropriate amount of type II seed crystals; maintaining the temperature at 20°C and stirring, filtering, and drying to obtain crystal form II.

[0025] The crystal form III described in this invention, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 5.6±0.2°, 6.0±0.2°, 12.0±0.2°, 14.5±0.2°, 19.3±0.2°, 20.0±0.2°, and 20.5±0.2°.

[0026] Preferably, the crystal form III is subjected to Cu-Kα radiation, and X-ray powder diffraction, expressed in 2θ angles, shows characteristic peaks at 5.6±0.2°, 6.0±0.2°, 7.7±0.2°, 9.5±0.2°, 11.2±0.2°, 12.0±0.2°, 13.1±0.2°, 14.5±0.2°, 15.2±0.2°, 17.3±0.2°, 17.8±0.2°, 19.3±0.2°, 20.0±0.2°, 20.5±0.2°, 21.4±0.2°, 22.5±0.2°, 24.0±0.2°, 25.9±0.2°, 27.8±0.2°, and 28.9±0.2°.

[0027] More preferably, the crystal form III, using Cu-Kα radiation, is X-ray powder diffracted at angles of 2θ at 5.6±0.2°, 6.0±0.2°, 7.7±0.2°, 9.5±0.2°, 10.0±0.2°, 11.2±0.2°, 12.0±0.2°, 13.1±0.2°, 14.5±0.2°, 15.2±0.2°, 16.9±0.2°, and 17.3°. Characteristic peaks are observed at ±0.2°, 17.8±0.2°, 18.1±0.2°, 18.7±0.2°, 19.3±0.2°, 20.0±0.2°, 20.5±0.2°, 21.4±0.2°, 22.5±0.2°, 24.0±0.2°, 24.4±0.2°, 25.9±0.2°, 26.8±0.2°, 27.8±0.2°, and 28.9±0.2°.

[0028] More preferably, the crystal form III has an X-ray powder diffraction pattern as shown in FIG29;

[0029] Most preferably, the crystal form III is irradiated with Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angles, is shown in Figure 29 with an error range of ±0.2°.

[0030] The differential scanning calorimetry curve of crystal form III shows an endothermic peak at 199.1℃±3℃.

[0031] Preferably, the DSC spectrum of crystal form III is shown in Figure 28.

[0032] The preparation method of the crystal form III includes the following steps: adding the free base of the compound of formula I and succinate in a certain equivalent ratio to an organic solvent, adjusting to a certain temperature, stirring until dissolved and clear; keeping warm for a certain time, cooling and keeping warm for a certain time, filtering and drying to obtain crystal form III;

[0033] Wherein, the equivalence ratio of the free base of Formula I compound to succinic acid is 1:0.5-2.0, preferably 1:0.8-1.2, and most preferably 1:1;

[0034] The organic solvent is selected from one or two of anisole, ethanol, diethyl ether, dichloromethane, tetrahydrofuran, toluene, ethyl acetate, acetone, methanol, and N,N-dimethylformamide; preferably, the organic solvent is selected from tetrahydrofuran.

[0035] The temperature adjustment is 60-80℃, preferably 65℃;

[0036] The heat preservation time is 1-4 hours, preferably 2 hours.

[0037] The cooling temperature is reduced to 5-20℃, preferably to 10℃;

[0038] The cooling and heat preservation time is 1-4 hours, preferably 1 hour.

[0039] Preferably, the preparation method of crystal form III of the present invention comprises the following steps: adding the free base of compound I and succinic acid to THF in a 1:1 equivalent ratio and stirring, adjusting the temperature to 65°C, stirring until dissolved and clear, and keeping warm for 2 hours; cooling to 10°C and keeping warm for 1 hour, filtering, and obtaining crystal form III.

[0040] The third aspect of this invention:

[0041] A pharmaceutical composition comprising a salt of a compound of formula I or a crystalline form thereof, the pharmaceutical composition may further include, as needed, a drug-received carrier, and is prepared into an oral pharmaceutical formulation, such as a tablet, capsule, injection, topical preparation, etc., as required for administration.

[0042] Preferably, it includes (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

[0043] More preferably, it includes the crystal form of (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

[0044] Particularly preferred crystal form II or crystal form III is selected from (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

[0045] The following is an explanation and description of the terminology used in this invention:

[0046] The chemical name of the compound of formula I is (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl) methyl ketone, and its structure is as follows:

[0047] The compound of formula I can be prepared by the preparation methods disclosed in the prior art, or by the preparation methods of similar compounds disclosed in the prior art.

[0048] "Therapeutic effective amount" refers to the amount of a compound that causes physiological or medical translation in an tissue, system, or subject. This amount is sought, including the amount of a compound, when applied to a subject, sufficient to prevent the occurrence of one or more symptoms of the treated disease or condition or to alleviate them to some extent.

[0049] The crystal structure of the present invention can be analyzed using various analytical techniques known to those skilled in the art, including but not limited to X-ray powder diffraction (XRD) and differential scanning calorimetry (DSC).

[0050] It is understood that the numerical values ​​described and protected in this invention are approximate. Variations within these values ​​may be attributed to equipment calibration, equipment errors, crystal purity, crystal size, sample size, and other factors.

[0051] It is understood that the crystal forms of the present invention are not limited to those that are exactly the same as the characteristic spectra described in the accompanying drawings, such as XRD and DSC. Any crystal form that has a characteristic spectra that are substantially the same or essentially the same as those described in the accompanying drawings falls within the scope of the present invention.

[0052] The X-ray powder diffraction or DSC patterns disclosed in this invention, and those substantially the same, also fall within the scope of this invention.

[0053] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0054] Beneficial technical effects of the present invention:

[0055] 1. This invention obtains 12 salts of the compound of formula I, including hydrochloride, phosphate, methanesulfonate, hydrobromide, sulfate, p-benzenesulfonate, oxalate, maleate, fumarate, succinate, tartrate, and citrate; completes tests on the crystal form, solubility, apparent hygroscopicity, powder properties, and solution stability of the obtained salts, and determines the dominant salt type, succinate; completes solid-state stability tests on the two dominant salt types, and both salt types maintain crystalline phase stability under accelerated stability test conditions.

[0056] 2. The free form of the compound (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone has a water solubility of less than 0.85 μg / ml, which is unfavorable for salt form development, crystallization process development, and formulation development. The compound successfully prepared by this invention has a superior salt form, and the two crystal forms II and III of the succinate have water solubilities of 145 mg / ml and 84.22 mg / ml, respectively, which are far greater than 0.85 μg / ml, thus solving the problems of salt form selection and difficulties in crystallization process development.

[0057] 3. This invention also solves the problems of mixed crystals, residual dissolution, difficulty in quality control, and poor powder properties in the development of crystal form II in solid form, and successfully prepares a single, stable, and qualified crystal form. Attached Figure Description

[0058] Figure 1: PXRD pattern of the hydrochloride of compound I prepared in Example 1;

[0059] Figure 2: PXRD pattern of the phosphate of compound I prepared in Example 1;

[0060] Figure 3: PXRD pattern of methanesulfonate of Formula I prepared in Example 1;

[0061] Figure 4: PXRD pattern of hydrobromide of Formula I prepared in Example 1;

[0062] Figure 5: PXRD pattern of the sulfate of compound I prepared in Example 1;

[0063] Figure 6: PXRD pattern of oxalate of Formula I prepared in Example 2;

[0064] Figure 7: PXRD pattern of maleate of formula I prepared in Example 2;

[0065] Figure 8: PXRD pattern of fumarate of Formula I prepared in Example 2;

[0066] Figure 9: PXRD pattern of succinate of Formula I prepared in Example 2;

[0067] Figure 10: PXRD pattern of the tartrate salt of Formula I prepared in Example 2;

[0068] Figure 11: PXRD pattern of citrate of Formula I prepared in Example 2;

[0069] Figure 12: PXRD pattern of p-methylbenzenesulfonate of Formula I prepared in Example 2 (amorphous morphology);

[0070] Figure 13: PXRD patterns of compound I p-benzenesulfonate before and after three days of storage;

[0071] Figure 14: PXRD patterns of oxalate, a compound of Formula I, before and after three days of storage;

[0072] Figure 15: PXRD patterns of maleate of Formula I before and after three days of storage.

[0073] Figure 16: PXRD patterns of the formula I compound fumarate before and after three days of storage.

[0074] Figure 17: PXRD patterns of succinate, a compound of Formula I, before and after three days of storage;

[0075] Figure 18: PXRD patterns of compound I tartrate before and after three days of storage;

[0076] Figure 19: PXRD patterns of citrate of Formula I before and after three days of storage;

[0077] Figure 20: PXRD patterns of compound I hydrochloride before and after three days of storage;

[0078] Figure 21: PXRD patterns of phosphate of Formula I before and after three days of storage;

[0079] Figure 22: PXRD patterns of compound I methanesulfonate before and after three days of storage;

[0080] Figure 23: PXRD patterns of compound I hydrobromide before and after three days of storage;

[0081] Figure 24: PXRD images of the sulfate of Formula I before and after three days of storage;

[0082] Figure 25: TG-DSC diagram of succinate II of Formula I;

[0083] Figure 26: XPRD diagram of succinate II of Formula I;

[0084] Figure 27: TG-DSC diagram of the mixed crystal form of compound I succinate;

[0085] Figure 28: TG-DSC diagram of succinate III of Formula I;

[0086] Figure 29: XPRD diagram of succinate III of Formula I;

[0087] Figure 30: 1H NMR spectrum of compound I, succinate. Detailed Implementation

[0088] The present invention is further illustrated by the following examples, but these are not intended to limit the invention.

[0089] Example 1: Salt type screening of inorganic acids of Formula I

[0090] Experimental objective: To screen salt types using hydrochloric acid, phosphoric acid, methanesulfonic acid, hydrobromic acid, and sulfuric acid as counterions.

[0091] Experimental method: Reaction crystallization method was used. A certain amount of API was weighed, dissolved in 1 mL of solvent, and a counterion solution was added. The mixture was stirred at 600 rpm for 24 h at room temperature. The resulting solid was filtered, dried under vacuum, and then subjected to PXRD testing.

[0092] Table 1. Experimental parameters and results for the salt forms of inorganic acids of Formula I.

[0093] Experimental results: Hydrochloride, phosphate, methanesulfonate, hydrobromide, and sulfate of compound I were obtained in THF, dichloromethane / ethanol (1:3), dichloromethane, dichloromethane, or THF, dichloromethane / ethanol (1:3), respectively. The sulfate was in an amorphous state, while the others were crystalline. Specific experimental parameters and results are shown in Table 1.

[0094] Figure 1 shows the PXRD pattern of the hydrochloride salt of Formula I obtained from experiment 230926-1; Figure 2 shows the PXRD patterns of the phosphate salt of Formula I obtained from experiments 230927-3 and 230922-2; experiment 230927-3 was obtained by suspension in dichloromethane / ethanol (1:3) and is in a crystalline state; experiment 230922-2 was obtained by suspension in dichloromethane and is in an amorphous state. Figure 3 shows the PXRD pattern of the methanesulfonate salt of Formula I obtained from experiment 230922-3; Figure 4 shows the PXRD patterns of the hydrobromide salt of Formula I obtained from experiments 230926-4 and 230921-4, and the hydrobromide salts obtained in dichloromethane and THF, respectively, have the same crystalline phase. Figure 5 shows the PXRD pattern of the sulfate of compound I obtained in experiment 230927-5. Experiment 230927-5 was obtained by suspension in dichloromethane / ethanol (1:3) and was in an amorphous state.

[0095] Example 2: Salt type screening of organic acids of Formula I

[0096] Experimental objective: To screen salt types using p-benzylsulfonic acid, oxalic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, and citric acid as counterions.

[0097] Experimental Method: Reaction crystallization was employed. A mixture of 1 mL dichloromethane and ethanol or ethyl acetate (volume ratio 1:3) was used as the solvent. Compound I and an organic acid were added separately until nearly saturated. Then, a stoichiometric ratio of compound I and an organic acid (1:1 ratio) was added, and the mixture was stirred at 600 rpm for 24 h at room temperature. The resulting solid was filtered, vacuum dried, and then subjected to PXRD analysis.

[0098] Table 2 shows the experimental parameters and results for the salt type screening of compound I, organic acid 9.

[0099] Experimental Results: Compounds of Formula I, including p-benzenesulfonate, oxalate, maleate, fumarate, succinate, tartrate, and citrate, were obtained. Among them, p-benzenesulfonate was in an amorphous state, while the other salts were crystalline. Specific experimental parameters and results are shown in Table 2.

[0100] Figure 6 shows the PXRD pattern of oxalate, a compound of formula I prepared in experiment 230928-2; Figure 7 shows the PXRD pattern of maleate, a compound of formula I prepared in experiment 230928-3; Figure 8 shows the PXRD pattern of fumarate, a compound of formula I prepared in experiment 230928-4; Figure 9 shows the PXRD pattern of succinate, a compound of formula I prepared in experiment 230928-5; Figure 10 shows the PXRD pattern of tartrate, a compound of formula I prepared in experiment 231009-1. Experiment 230928-6 was obtained by screening with a dichloromethane / ethanol mixed solvent and was amorphous; experiment 231009-1 used dichloromethane / ethyl acetate for screening, but did not yield tartrate with better crystallinity. Figure 11 shows the PXRD pattern of citrate, a compound of formula I prepared; experiment 230928-7 was obtained by screening with a dichloromethane / ethanol mixed solvent and had low crystallinity. Experiment 231009-3, prepared by suspension in dichloromethane / ethyl acetate, showed improved crystallinity. Figure 12 shows the PXRD pattern of p-toluenesulfonate, compound of formula I, prepared by experiment 231009-2, which is in an amorphous state.

[0101] Example 3: Characterization of the water solubility, apparent hygroscopicity, and powder properties of the salt form of compound I.

[0102] Experimental objective: To test the solubility, apparent hygroscopicity, and powder properties of the obtained salts in water.

[0103] Experimental method: ① Water solubility: Take 1 mL of pure water into an EP tube, add the sample, sonicate for about 1 min. If the solution is clear, continue to add the sample until it is insoluble, and record the solubility.

[0104] ② Evaluation of apparent hygroscopicity and preliminary stability: After weighing, the samples were placed in a room temperature and atmospheric environment for three days. The mass and crystal phase changes were measured, and the appearance changes of the samples were observed. The room temperature was 23℃, and the humidity was 43%. The specific experimental results are shown in Table 3, and the PXRD patterns before and after three days are shown in the attached figure.

[0105] Table 3 Results of water solubility, apparent hygroscopicity, and powder properties

[0106] The PXRD patterns of the 12 salt forms before and after three days of storage are shown below. Figure 13 shows the PXRD pattern of compound p-benzenesulfonate (Experiment No. 231014-1) before and after three days of storage; Figure 14 shows the PXRD pattern of oxalate (Experiment No. 231014-2) before and after three days of storage; Figure 15 shows the PXRD pattern of maleate (Experiment No. 231014-3) before and after three days of storage; Figure 16 shows the PXRD pattern of fumarate (Experiment No. 231014-4) before and after three days of storage; Figure 17 shows the PXRD pattern of succinate (Experiment No. 231014-5) before and after three days of storage. Figure 18 shows the PXRD patterns of compound I (tartrate) from experiment 231014-6 before and after three days of storage; Figure 19 shows the PXRD patterns of compound I (citrate) from experiment 231014-7 before and after three days of storage; Figure 20 shows the PXRD patterns of compound I (hydrochloride) from experiment 231014-8 before and after three days of storage; Figure 21 shows the PXRD patterns of compound I (phosphate) from experiment 231014-9 before and after three days of storage; Figure 22 shows the PXRD patterns of compound I (methanesulfonate) from experiment 231014-10 before and after three days of storage, where the crystal form changes after three days, showing the peak of the active pharmaceutical ingredient, indicating instability; Figure 23 shows the PXRD patterns of compound I (hydrobromide) from experiment 231014-11 before and after three days of storage, where the crystal form changes after three days, the crystal phase differs from the active pharmaceutical ingredient, and the crystal phase is unstable. Figure 24 shows the PXRD patterns of the sulfate of Formula I compound 231014-12 before and after three days of storage.

[0107] From the data in Table 3 above, we can see that: after the samples were placed in an environment of room temperature 23℃ and humidity 43% for three days, (1) appearance changes: the appearance of fumarate changed from light yellow flaky powder to yellow flaky powder, the appearance of methanesulfonate and hydrobromide did not change but the viscosity increased, and the appearance and viscosity of other salt types did not change. (2) hygroscopicity: the solid weight of methanesulfonate increased by 1.2 mg after three days, which is 2.4%, so methanesulfonate is hygroscopic; after the other salt types were placed in an environment of room temperature 23℃ and humidity 43% for three days, the weight increase was less than 2%, so the other salt types are slightly hygroscopic or almost non-hygroscopic. (3) after the methanesulfonate and hydrobromide were placed for three days, the raw material peak and crystal phase changed respectively, and the stability was poor, while the crystal phase of other salt types did not change. (4) Powder properties: Oxalate, maleate, hydrobromide, hydrochloride and fumarate have good flowability, low static electricity and do not stick to the wall; succinate and citrate have moderate flowability, static electricity and stick to the wall; phosphate and methanesulfonate have poor flowability, no static electricity and stick to the wall.

[0108] Based on Examples 1, 2, and 3, it is evident that sulfate and p-benzenesulfonate failed to form crystals. Methanesulfonate and hydrobromide showed changes in their raw material peaks and crystal phases after three days of storage, indicating poor stability. Tartrate showed hygroscopicity and significant weight gain after three days of storage. Therefore, the five salt forms—sulfate, p-benzenesulfonate, methanesulfonate, hydrobromide, and tartaric acid—are not suitable for further development.

[0109] Example 4: Chemical stability test of compound I salt solution

[0110] Experimental method: Prepare the test solution and perform related substance tests on days 0, 1, 2 and 3 respectively.

[0111] Table 4. Preparation of test solution and HPLC methods

[0112] Table 5. Stability results after 3 days of storage.

[0113] Experimental Results: After 3 days in aqueous solution, no significant changes were observed in the relevant substances of the twelve API salts. However, as shown in Table 5, during the salt formation process, the impurity purity at RT17.5 min increased to 2.10%, 0.20%, 0.21%, and 0.56%, respectively, which is 10-105 times higher than the free state of 0.02%. During the salt formation process, the impurity purity at RT12.7 min for phosphate was 0.43%, an increase of 0.13% compared to 0.30%. In summary, the aqueous solutions of oxalate, fumarate, phosphate, hydrochloride, and citrate exhibited poor stability, while the aqueous solutions of maleate and succinate showed good stability.

[0114] Example 5: Solid-state stability test of maleate and succinate of Formula I

[0115] 1. Experimental objective: To test the chemical stability of maleate and succinate under accelerated stability conditions.

[0116] 2. Experimental method: Accurately weigh about 0.1g of solid sample and place it at 40℃±2℃ and 75%±5% relative humidity. Prepare a 1mg / mL solution on days 0, 3 and 7 for related substance testing. The results are shown in Table 6 below.

[0117] Table 6 Solid-state stability test

[0118] Qualitatively, after 0, 3, and 7 days of storage, the impurities did not increase significantly. When the maleate compound was prepared into a solution at 40℃±2℃ and 75%±5% relative humidity, the impurity content at 12.7 min (RT) increased from 0.03% to 0.22% after 0, 3, and 7 days of storage. Therefore, succinate is stable after 7 days of storage at 40℃±2℃ and 75%±5% relative humidity, while maleate will undergo degradation under these conditions, with the impurity at 12.7 min (RT) showing a gradual increasing trend.

[0119] In summary, all 12 salt forms have different types of problems. Succinate has stable crystal phase, low hygroscopicity, and good stability in aqueous solution and solid state compared to other salt forms. Its problems are slightly less than those of other salt forms. Therefore, we will try to develop a crystal form for it.

[0120] Example 6: Preparation of Succinate Crystal Form II of Compound I and Detection of Residual Solvent

[0121] Crystallization temperature screening experiment: DCM and EtOH were used as solvents, and crystallization was carried out at 0-50℃ respectively. The precipitated crystals were then detected by TG-DSC. The results are shown in Table 7 below.

[0122] Table 7 Screening of crystallization temperatures for succinate crystal form II

[0123] During crystal preparation, when the crystallization temperature is above 30℃, the resulting solid is identified as a mixed crystal by TG-DSC; while when the crystallization temperature is below or equal to 20℃, the resulting solid is identified as API; only when the temperature is between 20-30℃ does a new, single crystal form appear, as shown in Figure 25 below. Figure 25 shows that thermal analysis reveals an endothermic peak at 172.76℃ (peak value), indicating a new crystal form, named crystal form II. X-ray powder diffraction of crystal form II, measured using Cu-Kα rays, shows diffraction peaks at 2θ angles (in °): 5.3±0.2°, 10.6±0.2°, 15.1±0.2°, 16.1±0.2°, 18.3±0.2°, and 21.1±0.2°.

[0124] The residual solvent was detected in the crystal form II prepared in this embodiment, and the residual data are shown in Table 8.

[0125] Table 8. Detection of Residual Solvents in Succinate Crystal Form II

[0126] Residual Experiment Conclusion: The succinate crystal form II prepared in this example contains approximately 0.2% dichloromethane solvent residue, which exceeds the specified limit by more than 2 times. (ICH Guideline Q3C(R8) stipulates that dichloromethane is a Class 2 solvent that should be restricted, with a concentration limit of 600 ppm. Therefore, the dichloromethane solvent residue concentration of crystal form II does not comply with the ICH Guideline Q3C(R8).

[0127] Example 7: Preparation of Succinate Crystal Form II of Formula I and Detection of Residual Solvent

[0128] Compound I and succinic acid were added to anisole in a 1:1 equivalence ratio and stirred. The temperature was adjusted to 30°C and stirred until dissolved and clear. The temperature was lowered to 20°C and kept warm for 1 hour. An ethanol solution of succinic acid was added. Seed crystal II, which was 10% of the mass of the free state, was added. The mixture was kept warm at 20°C and stirred. After filtration, a pale yellow solid was obtained, which is crystal form II.

[0129] The obtained solid was subjected to TG-DSC analysis, and the differential scanning calorimetry chromatogram of the solid obtained at a heating rate of 10℃ / min was consistent with that of Figure 25 obtained in Example 6.

[0130] The X-ray powder diffraction of crystal form II, measured using Cu-Kα rays, shows diffraction peaks at 2θ angles (in °): 5.3±0.2°, 10.6±0.2°, 15.1±0.2°, 16.1±0.2°, 18.3±0.2°, and 21.1±0.2°. Specific diffraction data for crystal form II of compound I (succinate) are shown in Table 9; Figure 26 shows the XPRD pattern of crystal form II of succinate.

[0131] Table 9. Diffraction data of succinate crystal form II of compound I.

[0132] Residual solvent was detected in crystal form II, and the data are shown in Table 10.

[0133] Table 10 Residual Solvent Detection for Crystal Form II

[0134] Residual solvent test results: All residual solvent indicators of the succinate crystal form II prepared in this example meet the requirements of ICH guideline Q3C(R8).

[0135] Example 8: Preparation of succinate sample of Formula I (crystal form II)

[0136] Under nitrogen protection, 50 ml of anisole was added; stirring was started, followed by 20 ml of ethanol and 10 g of compound I; the temperature was adjusted to 20-30 °C; stirring was continued until dissolved and clear; a solution of 4.3 g of succinic acid in 130 ml of ethanol was added; 1.2 g of seed crystals was added; the mixture was kept at 20 °C and stirred for at least 6 hours, filtered, and the filter cake was washed with 10 ml of EA. The mixture was then vacuum dried at 40 °C for 6 hours to obtain 9.15 g of a pale yellow solid, with a yield of 75.30%. The X-ray powder diffraction pattern obtained using Cu-Kα rays was consistent with succinate crystal form II in Example 7, and TG-DSC analysis of the obtained solid confirmed its consistency with succinate crystal form II.

[0137] Example 9: Solid-state stability test of succinate of Formula I crystal form II

[0138] Succinate crystal form II was placed at 40℃±5℃, 60℃±5℃ and relative humidity of 90%±5% for 30 days and analyzed by HPLC. The data are shown in Tables 11-13 below.

[0139] Table 11 Solid-state stability experimental data (placement conditions: 40℃±2℃)

[0140] Table 12 Solid-state stability experimental data (placement conditions: 60℃±2℃)

[0141] Table 13 Solid-state stability experimental data (placement conditions: RH 92.5%)

[0142] Conclusion: Based on the stability data of succinate crystal form II of Formula I, it can be concluded that the impurities of succinate crystal form II of Formula I do not increase significantly under the conditions of 40℃, 60℃ and relative humidity of 92.5%, thus indicating that crystal form II has good solid-state stability.

[0143] Example 10: Hygroscopicity and solubility of succinate (Formula I)

[0144] Hygroscopicity test of succinate crystal form II: During the process of humidity change from 0 to 80°C at 25°C, the weight increase was less than 0.1%, and the data showed that crystal form II has almost no hygroscopicity.

[0145] The crystal form II sample was ground and sieved. 2 mL of ultrapure water was placed in an EP tube, and the sieved drug was added. The mixture was shaken for 30 seconds and sonicated for about 1 minute. If the solution was clear, the drug was added again until it was insoluble. The solubility of crystal form II was recorded as 145 mg / mL.

[0146] Example 11: Preparation of Succinate Crystal Form III of Compound I

[0147] Crystallization temperature screening experiment: Add compound of formula I: succinic acid = 1:1 (equivalent ratio) THF to the reactor and stir. Adjust the temperature to 25±5℃ and stir until dissolved and clear. Keep warm for 2 hours; cool down to 10℃ and keep warm for 1 hour. Filter to obtain an off-white solid.

[0148] The obtained solid was subjected to TG-DSC analysis, and the differential scanning calorimetry (DSC) curve of the solid was obtained at a heating rate of 10 °C / min, as shown in Figure 27. As can be seen from Figure 27 in the specification, the thermal analysis showed two endothermic peaks at 173.00 °C (peak) and 196.17 °C (peak), suggesting that the obtained solid is a mixed crystal of two crystalline forms. TG-DSC analysis revealed that the two crystalline forms of succinate obtained by the cooling method exhibited a mixed crystal problem.

[0149] This invention attempts to change the temperature and use 20-70℃ conditions to crystallize, and the precipitated crystals are detected by TG-DSC. The results are shown in Table 14 below.

[0150] Table 14 shows the crystal forms obtained under different crystallization problems.

[0151] Example 12: Preparation of Succinate Crystal Form III of Compound I

[0152] Compound I and succinic acid were added to THF in a 1:1 (equivalent ratio) mixture and stirred. The temperature was adjusted to 65°C, and the mixture was stirred until dissolved and clear. This mixture was kept at this temperature for 2 hours. The temperature was then lowered to 10°C and kept at this temperature for 1 hour. The mixture was filtered to obtain an off-white solid. The solid was subjected to TG-DSC analysis, and the differential scanning calorimetry (DSC) curve of the solid was obtained at a heating rate of 10°C / min, as shown in Figure 28. Figure 28 shows that the thermal analysis revealed an endothermic peak at 199.13°C (peak value), indicating a single crystal form, which was named succinate III of compound I.

[0153] X-ray powder diffraction (XPD) of succinate crystal form III, measured using Cu-Kα rays, shows diffraction peaks at 2θ angles (in °): 5.6±0.2°, 6.0±0.2°, 12.0±0.2°, 14.5±0.2°, 19.3±0.2°, 20.0±0.2°, and 20.5±0.2°. Diffraction angle data for succinate crystal form III of Formula I are shown in Table 15; Figure 29 shows the XPRD pattern of succinate crystal form III of Formula I.

[0154] Table 15. Diffraction data of succinate (formula I) crystal type III.

[0155] Example 13: Preparation of Succinate Crystal Form III of Compound I

[0156] Under nitrogen protection, 30 ml of THF was added; stirring was started, and 10 g of compound I was added; 4.3 g of succinic acid was added, and the temperature was adjusted to 65±5℃; stirring was continued until dissolved and clear, and the mixture was kept at this temperature for 2 hours; the mixture was then distilled under reduced pressure to 100 ml; stirring was continued, the temperature was adjusted to 60±5℃, and then cooled to 10±5℃ and kept at this temperature for 1 hour. The mixture was filtered, the filter cake was washed with 10 ml of THF, and then dried under vacuum at 40℃ for 6 hours to obtain 10.45 g of an off-white solid, with a yield of 86.00%. The obtained crystal form was consistent with crystal form III in Example 12.

[0157] Example 14: Solid-state stability of succinate of Formula I (crystal form III)

[0158] Solid-state stability of crystal form III was tested. Succinate crystal form III was placed at 40℃±2℃, 60℃±2℃ and relative humidity of 90%±5% for 30 days, and its data were analyzed by HPLC. The data are shown in Tables 16, 17 and 18.

[0159] Table 16 Solid-state stability experimental data (placement conditions: 40℃±5℃)

[0160] Table 17 Solid-state stability experimental data (placement conditions: 60℃±5℃)

[0161] Table 18 Solid-state stability experimental data (placement conditions: RH 90% ± 5%)

[0162] The stability data of crystal form III show that the impurities in crystal form III do not increase significantly under the conditions of 40℃±5℃, 60℃±5℃ and relative humidity of 90%±5%, thus indicating that crystal form III has good solid-state stability.

[0163] Example 15: Hygroscopicity test of succinate crystal form III of Formula I.

[0164] Hygroscopicity test of crystal form III: During the process of changing humidity from 0 to 80% at 25℃, the weight increased by about 0.13%, indicating that crystal form III is not hygroscopic.

[0165] Example 16: Solubility test of succinate crystal form III of Formula I

[0166] The crystal form III sample was ground and sieved. 2 mL of ultrapure water was placed in an EP tube, and the sieved drug was added. The mixture was shaken for 30 seconds and sonicated for about 1 minute. If the solution was clear, the drug was added again until it was insoluble. The solubility of crystal form III was recorded as 84.22 mg / mL.

[0167] Example 17: Structural characterization and confirmation of succinate, a compound of formula I

[0168] Using a 1 mL mixture of dichloromethane and ethanol (volume ratio 1:3) as the solvent, compound I and succinic acid were added separately until nearly saturated. Then, compound I and succinic acid in a stoichiometric ratio of 1:1 were added, and the mixture was stirred at 600 rpm for 24 h at room temperature. The resulting solid was filtered, dried under vacuum, and characterized by NMR. The NMR assignments are shown in Table 19 below, and the NMR spectra are shown in Figure 30.

[0169] Table 19 shows the NMR assignments of succinate, a compound of formula I.

[0170] According to Table 19, the chemical shifts are 10.55 with two singlets and 2.64 with four singlets. These two positions represent succinic acid, while the other positions represent the free state. The ratio of free state to succinic acid in compound I is 1:1, as determined by the number of H atoms.

[0171] Example 18: Determination of inhibitory effects on the proliferation of various cancer cells

[0172] The inhibitory effects of the compound on the proliferation of six cell lines, including human breast cancer cells MDA-MB-231 and MDA-MB-453, human prostate cancer cells DU145, human thyroid squamous cell carcinoma cells SW579, human ovarian cancer cells SKOV-3, and human colon cancer cells HT-29, were tested using the following methods.

[0173] Experimental steps

[0174] 1) Culture cells and passage them at least twice, harvest cells, adjust cell density, and seed them in 96-well plates: DU 145, MDA-MB-231, MDA-MB-453, SW579: 1000 cells / well; SKOV-3, HT-29: 800 cells / well; 2) The next day, after cell adhesion, dissolve the compound in sterile water and the positive drug in DMSO, dilute with their respective solvents to prepare drug stock solutions, and prepare (10×) drug working solution using complete culture medium; add 10 μL of test drug / well according to the cell arrangement diagram, set up 9 concentration gradients, starting at 20 μM, 3-fold serial dilutions, 3 replicates for each concentration, and set up negative control wells without drug and blank control wells containing only culture medium; 3) Place the 96-well plate in a CO2 incubator and incubate at 37°C for 120 hours; 4) After incubation, add an equal volume of cell culture medium to each well. 5) Mix with a fixed-track oscillator for 2 minutes to induce cell lysis; 6) Incubate the well plate at room temperature for 10 minutes to stabilize the luminescence signal; 7) Transfer the supernatant to a white opaque 96-well plate and record the luminescence signal using a microplate reader.

[0175] Experimental results

[0176] The test compound formula I, compound succinate crystal form II, and control drug were used to test the IC50 of MDA-MB-231, MDA-MB-453, DU145, SW579, SKOV-3, and HT-29 cells. 50 The values ​​are shown in Table 20 below, where the IC50 of the compounds is determined. 50 The categories are as follows, according to the instructions.

[0177] "+" indicates IC 50 The measured value is greater than 5 μM; "++" indicates IC50 value. 50 The measured value is less than or equal to 5 μM and greater than 2 μM; "++" indicates IC50 value. 50 The measured value is less than or equal to 2 μM and greater than 0.1 μM.

[0178] Table 20 shows the IC50 of succinate (Form I) and control drugs palbociclib and abecilib on cells. 50 Value Summary Table

[0179] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A pharmaceutically acceptable salt of compound I (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl) methyl ketone, The salt is selected from: hydrochloride, phosphate, methanesulfonate, hydrobromide, sulfate, p-benzenesulfonate, oxalate, maleate, fumarate, succinate, tartrate, and citrate.

2. The pharmaceutically acceptable salt according to claim 1, characterized in that: It is (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

3. Crystal form of compound (6-((4-(2-(diethylamino)benzo[d]thiazolyl-6-yl)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone succinate.

4. The crystal form according to claim 3, characterized in that: The crystal form is II. The X-ray powder diffraction of the crystal form II using Cu-Kα radiation, expressed in 2θ angles, has characteristic peaks at 5.3±0.2°, 10.6±0.2°, 15.1±0.2°, 16.1±0.2°, 18.3±0.2°, and 21.1±0.2°.

5. The crystal form according to claim 4, characterized in that, The crystal form II, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 5.3±0.2°, 10.6±0.2°, 14.0±0.2°, 15.1±0.2°, 16.1±0.2°, 16.9±0.2°, 18.3±0.2°, 19.5±0.2°, 21.1±0.2°, 23.8±0.2°, 25.5±0.2°, 26.8±0.2°, and 28.2±0.2°.

6. The crystal form according to claim 4, characterized in that, The crystal form II, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 2θ angles of 5.3±0.2°, 9.2±0.2°, 9.6±0.2°, 10.6±0.2°, 12.0±0.2°, 13.4±0.2°, 14.0±0.2°, 15.1±0.2°, 16.1±0.2°, 16.9±0.2°, 18.3±0.2°, 18.5±0.2°, 19.5±0.2°, 21.1±0.2°, 22.7±0.2°, 23.3±0.2°, 23.8±0.2°, 24.5±0.2°, 25.5±0.2°, 26.8±0.2°, 28.2±0.2°, and 32.2±0.2°.

7. The crystal form according to claim 4, characterized in that, The differential scanning calorimetry curve of crystal form II has an endothermic peak at 172.8±3℃.

8. A method for preparing the crystal form according to claim 4, characterized in that: The free base of compound I and succinate were added to an organic solvent in a certain equivalence ratio, the temperature was adjusted, and the mixture was stirred until dissolved and clear. The mixture was then cooled and kept at that temperature for a certain time. Next, an organic solvent containing succinic acid was added, followed by seed crystal II. The mixture was kept at that temperature and stirred, then filtered and dried to obtain crystal form II. Wherein, the equivalence ratio of the free base of Formula I compound to succinic acid is 1:0.5-2.0, preferably 1:0.8-1.2, and most preferably 1:1; The organic solvent is selected from one or two of anisole, ethanol, diethyl ether, dichloromethane, tetrahydrofuran, toluene, ethyl acetate, acetone, methanol, and N,N-dimethylformamide; preferably, the organic solvent is selected from one or two of anisole and ethanol. The temperature adjustment is 20-30℃, preferably 30℃; The insulation temperature is 20-30℃, preferably 20℃; The heat preservation time is 0.5-2 hours, preferably 1 hour.

9. The preparation method according to claim 8, characterized in that: Compound of Formula I and succinic acid were added to anisole at an equivalent ratio of 1:1 and stirred. The temperature was adjusted to 30°C and stirred until dissolved and clear. The temperature was lowered to 20°C and kept warm for 1 hour. An ethanol solution of succinic acid was added. An appropriate amount of type II seed crystals was added. The mixture was kept at 20°C, stirred, filtered, and dried to obtain crystal form II.

10. The crystal form according to claim 3, characterized in that: The crystal form is III. The X-ray powder diffraction of the crystal form III using Cu-Kα radiation, expressed in 2θ angles, has characteristic peaks at 5.6±0.2°, 6.0±0.2°, 12.0±0.2°, 14.5±0.2°, 19.3±0.2°, 20.0±0.2° and 20.5±0.2°.

11. The crystal form according to claim 10, characterized in that, The crystal form III, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 2θ angles of 5.6±0.2°, 6.0±0.2°, 7.7±0.2°, 9.5±0.2°, 11.2±0.2°, 12.0±0.2°, 13.1±0.2°, 14.5±0.2°, 15.2±0.2°, 17.3±0.2°, 17.8±0.2°, 19.3±0.2°, 20.0±0.2°, 20.5±0.2°, 21.4±0.2°, 22.5±0.2°, 24.0±0.2°, 25.9±0.2°, 27.8±0.2°, and 28.9±0.2°.

12. The crystal form according to claim 10, characterized in that, Crystal form III, using Cu-Kα radiation, was observed to undergo X-ray powder diffraction at angles of 2θ at 5.6±0.2°, 6.0±0.2°, 7.7±0.2°, 9.5±0.2°, 10.0±0.2°, 11.2±0.2°, 12.0±0.2°, 13.1±0.2°, 14.5±0.2°, 15.2±0.2°, 16.9±0.2°, and 17.3±0.2°. Characteristic peaks are observed at 2°, 17.8±0.2°, 18.1±0.2°, 18.7±0.2°, 19.3±0.2°, 20.0±0.2°, 20.5±0.2°, 21.4±0.2°, 22.5±0.2°, 24.0±0.2°, 24.4±0.2°, 25.9±0.2°, 26.8±0.2°, 27.8±0.2°, and 28.9±0.2°.

13. The crystal form according to claim 10, characterized in that, The differential scanning calorimetry curve of crystal form III shows an endothermic peak at 199.1℃±3℃.

14. A method for preparing the crystal form according to claim 10, characterized in that: The free base of compound I and succinate were added to an organic solvent in a certain equivalence ratio, the temperature was adjusted to a certain level, and the mixture was stirred until dissolved and clear. The mixture was kept at the temperature for a certain time, cooled down and kept at the temperature for a certain time, and then filtered and dried to obtain crystal form III. Wherein, the equivalence ratio of the free base of Formula I compound to succinic acid is 1:0.5-2.0, preferably 1:0.8-1.2, and most preferably 1:1; The organic solvent is selected from one or two of anisole, ethanol, diethyl ether, dichloromethane, tetrahydrofuran, toluene, ethyl acetate, acetone, methanol, and N,N-dimethylformamide; preferably, the organic solvent is selected from tetrahydrofuran. The temperature adjustment is 60-80℃, preferably 65℃; The heat preservation time is 1-4 hours, preferably 2 hours. The cooling temperature is reduced to 5-20℃, preferably to 10℃; The cooling and heat preservation time is 1-4 hours, preferably 1 hour.

15. The preparation method according to claim 14, characterized in that: Add the free base of compound I and succinic acid to THF in a 1:1 equivalent ratio and stir. Adjust the temperature to 65°C and stir until dissolved and clear. Keep warm for 2 hours. Cool to 10℃ and hold for 1 hour, then filter to obtain crystal form III.

16. A pharmaceutical composition comprising a salt of the compound of formula I according to claim 1 or a crystal form thereof.

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