Crystalline forms of casein kinase 1 epsilon inhibitors, methods of preparation and uses thereof

Abstract

The present invention discloses a crystalline form of a casein kinase 1ε inhibitor, a method for preparing the crystalline form, and uses thereof. Specifically, the present invention relates to a crystalline form of (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate, a method for preparing the crystalline form, and uses thereof. The crystalline form of the casein kinase 1ε inhibitor compound obtained by the present invention has good aqueous solubility and excellent oral absorption, making it suitable for use in the treatment of malignant tumors and autoimmune diseases that are effective through the inhibition of casein kinase 1ε activity.

Description

[Technical field]

[0001] The present invention relates to the process of chemical drug crystal forms and drug manufacturing technology, in particular to the crystal form of compound (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate, the preparation method of the crystal form and its use. [Background technology]

[0002] Protein kinases play an important role in signal transduction, mediating intracellular signal transduction through phosphorylated substrate proteins and being important drug targets. Casein kinase 1 (CK1) belongs to the serine-threonine kinase family, and in mammals there are seven subtypes: α, β, γ1, γ2, γ3, δ and ε. These subtypes are involved in Wnt signal transduction, circadian rhythm, cell signal transduction, membrane transport, DNA replication, DNA damage and RNA metabolism, and can regulate biological functions.

[0003] Casein kinase 1ε (CK1ε), an important subtype in the CK1 family, is a key regulator of the proliferation and survival processes of various cells, which not only plays an important role in regulating the circadian rhythm, but also plays an important role in the development and progression of cancer. For example, suppressing CK1ε by drugs or silencing CK1ε by shRNA mediation inhibits the proliferation and survival of various cancers, including pancreatic cancer, sarcoma, breast cancer, rectal cancer, ovarian cancer, leukemia, etc. Research has shown that the mechanism of action of CK1ε regulation on tumor proliferation and survival is still unclear because the CK1 gene has no substrate specificity, and may be related to Akt, MYC, β-catenin, etc. (Varghese et al., Scientific Reports, 2018, 8:13621). In addition, CK1ε, as an important component of regulating the Wnt signaling pathway, may promote the onset and progression of chronic lymphocytic leukemia (CLL). CK1ε is significantly upregulated in CLL patients, and suppression of CK1ε can inhibit CLL progression but has no effect on the BCR-related pathway. Moreover, CK1ε can enhance the antitumor activity of ibrutinib, a BTK inhibitor of the BCR pathway, in vivo and in vitro.

[0004] Therefore, CK1ε is also an important antitumor target (Janovska et al., Blood, 2019, 11:1206-1218). Therefore, the development of drugs that have the function of inhibiting CK1ε can exert important therapeutic effects on various diseases, such as cancer and autoimmune diseases.

[0005] The phenomenon that the same drug molecule forms multiple crystal forms is called drug polymorphism, which is widespread in the process of drug development and is an inherent characteristic of organic small molecule compounds. Different crystal forms may have differences in melting point, solubility, dissolution performance, chemical stability, etc., and their physical and chemical properties directly affect the safety and efficacy of drugs. Therefore, the study and control of crystal forms is an important research content in the process of drug research and development. Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention provides a crystalline form of a casein kinase 1 epsilon inhibitor of formula (I), i.e. (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate, a method for preparing the crystalline form and uses thereof. JPEG0007603292000001.jpg92136

[0007] The compound represented by formula (I) of the present invention, i.e., (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate, is a novel casein kinase 1ε (CK1ε) inhibitor and has strong in vitro CK1ε kinase inhibitory effect, in vitro antitumor cell activity and in vivo antitumor activity. Therefore, by conducting research on the relevant crystal forms of the compound, it is expected that the compound can be better applied in clinical practice.

[0008] A series of crystalline products of the compound of formula (I) obtained under different crystallization conditions were subjected to powder X-ray diffraction measurements, and different crystalline forms, A, B, C, D, E, F and I, were discovered. Correlative studies on the aqueous solubility, stability and pharmacokinetics of these crystalline forms when administered orally to rats and beagle dogs were carried out. [Means for solving the problem]

[0009] The technical means of the present invention are as follows: The phosphatidylinositol-3-kinase inhibitor provided by the present invention is a compound represented by formula (I): JPEG0007603292000002.jpg92136Has different crystal forms A, B, C, D, E, F, and I. Furthermore,

[0010] Crystal form A of the compound of formula (I) is obtained by irradiating crystal form A with Cu-Kα to obtain a powder X-ray diffraction pattern expressed as 2θ angles, which has characteristic peaks at 7.848±0.2°, 11.618±0.2°, 15.562±0.2°, 15.853±0.2°, 20.185±0.2°, and 25.655±0.2°.

[0011] A preferred embodiment of crystalline form A has characteristic peaks at 11.926±0.2°, 15.271±0.2°, 20.185±0.2°, 21.851±0.2°, and 24.799±0.2°. A more preferred embodiment of crystalline form A has characteristic peaks at 8.531±0.2°, 10.730±0.2°, 15.176±0.2°, 22.826±0.2°, and 25.369±0.2°. A further preferred embodiment of crystalline form A has characteristic peaks at 23.506±0.2° and 25.369±0.2°.

[0012] In a more preferred embodiment of crystalline form A, it simultaneously has characteristic peaks at 5.005±0.2°, 10.000±0.2°, 18.185±0.2°, 19.499±0.2°, 23.226±0.2°, and 27.070±0.2°. As a more specific preferred embodiment of the crystalline form A, said crystalline form A has characteristic peak data shown in Table 1 of the Examples. In a more specific preferred embodiment of crystalline form A, said crystalline form A has a powder X-ray diffraction pattern as shown in FIG.

[0013] Crystal form B of the compound of formula (I) is irradiated with Cu-Kα to obtain a powder X-ray diffraction pattern expressed as 2θ angles, which has characteristic peaks at 10.739±0.2°, 11.961±0.2°, 13.726±0.2°, 14.374±0.2°, 23.714±0.2°, 24.057±0.2°, and 25.033±0.2°.

[0014] A preferred embodiment of crystalline form B simultaneously has characteristic peaks at 14.809±0.2°, 18.574±0.2°, 22.313±0.2°, 26.125±0.2° and 26.659±0.2°.

[0015] A more preferred embodiment of crystalline form B has characteristic peaks at 9.837±0.2°, 18.078±0.2°, 19.759±0.2°, 24.724±0.2°, 27.330±0.2°, and 29.863±0.2°.

[0016] In a more preferred embodiment, crystalline form B simultaneously has characteristic peaks at 26.125±0.2°, 26.659±0.2°, 27.330±0.2° and 29.863±0.2°. As a more specific preferred embodiment of the crystalline form B, said crystalline form B has characteristic peak data shown in Table 2 of the Examples. In a more specific preferred embodiment of crystalline form B, said crystalline form B has a powder X-ray diffraction pattern as shown in FIG.

[0017] The compound of formula (I) has crystalline form C, which has a powder X-ray diffraction pattern expressed as 2θ angles by irradiating the compound with Cu-Kα, and has characteristic peaks at 7.319±0.2°, 8.092±0.2°, 11.763±0.2°, 14.728±0.2°, 15.855±0.2°, and 16.265±0.2°. A preferred embodiment of crystalline form C has characteristic peaks at 7.965±0.2°, 9.103±0.2° and 13.338±0.2°. A more preferred embodiment of crystalline form C has characteristic peaks at 3.880±0.2°, 6.478±0.2°, and 19.613±0.2°. In a further preferred embodiment of crystalline form C, said crystalline form C has characteristic peak data as shown in Table 3 of the Examples. In a more specific preferred embodiment of crystalline form C, said crystalline form C has a powder X-ray diffraction pattern as shown in FIG.

[0018] The compound of formula (I) has crystalline form D, which has characteristic peaks at 11.308±0.2°, 17.237±0.2°, 18.568±0.2°, 20.213±0.2°, 21.148±0.2°, and 21.293±0.2° when irradiated with Cu-Kα.

[0019] As a preferred form of crystalline form D, it simultaneously has characteristic peaks at 10.341±0.2°, 13.819±0.2°, 17.982±0.2°, 20.804±0.2°, 22.729±0.2°, 25.035±0.2°, 25.289±0.2° and 28.008±0.2°. In a more preferred embodiment of crystalline form D, said crystalline form D has characteristic peak data as shown in Table 4 of the Examples. In a more preferred embodiment of crystalline form D, said crystalline form D has an X-ray powder diffraction pattern as shown in FIG.

[0020] Crystal form E of the compound represented by formula (I) is obtained by irradiating crystal form E with Cu-Kα, and the powder X-ray diffraction pattern expressed as 2θ angles has characteristic peaks at 7.114±0.2°, 7.936±0.2°, 9.973±0.2°, 11.040±0.2°, 14.484±0.2°, 15.504±0.2°, 16.507±0.2°, 20.071±0.2°, and 21.555±0.2°.

[0021] A preferred embodiment of crystalline form E has characteristic peaks at 5.492±0.2°, 22.519±0.2°, 24.464±0.2° and 27.980±0.2°.

[0022] A more preferred embodiment of crystalline form E has characteristic peaks at 11.531±0.2°, 18.158±0.2°, 26.576±0.2°, 26.723±0.2°, 26.979±0.2°, 28.903±0.2°, and 33.404±0.2°.

[0023] A preferred embodiment of crystalline form E simultaneously has characteristic peaks at 12.887±0.2°, 18.526±0.2°, 21.929±0.2° and 24.152±0.2°. In a further preferred embodiment of crystalline form E, said crystalline form E has characteristic peak data as shown in Table 5 of the Examples. In a more preferred embodiment of crystalline form E, said crystalline form E has a powder X-ray diffraction pattern as shown in FIG.

[0024] The compound of formula (I) has crystalline form F, which has characteristic peaks at 9.959±0.2°, 13.604±0.2°, 14.830±0.2°, 20.645±0.2°, 23.717±0.2°, and 26.262±0.2° when irradiated with Cu-Kα.

[0025] A preferred embodiment of crystalline form F simultaneously has characteristic peaks at 10.410±0.2°, 12.704±0.2°, 15.773±0.2° and 22.390±0.2°. In a more preferred embodiment of crystalline form F, said crystalline form F has characteristic peak data as shown in Table 6 of the Examples. In a more preferred embodiment of crystalline form F, said crystalline form F has an X-ray powder diffraction pattern as shown in FIG.

[0026] The compound of formula (I) has crystalline form I, which, when irradiated with Cu-Kα, has a powder X-ray diffraction pattern expressed as 2θ angles, having characteristic peaks at 7.321±0.2°, 7.538±0.2°, 12.317±0.2°, 14.643±0.2°, 15.839±0.2°, and 19.458±0.2°. A preferred embodiment of crystalline form I has characteristic peaks at 3.978±0.2° and 15.081±0.2°. In a more preferred embodiment of crystalline form I, said crystalline form I has characteristic peak data as shown in Table 7 of the Examples. In a more preferred embodiment of crystalline form I, said crystalline form I has a powder X-ray diffraction pattern as shown in FIG.

[0027] The present invention further provides a method for crystallizing different crystal forms of (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate. The specific method is as follows:

[0028] The preparation method of crystalline form A includes the steps of: Step 1, dissolving the compound of formula (I) in a suitable amount of solvent I by heating, selectively filtering the resulting clear solution while hot, and cooling the resulting solution for crystallization, the solvent I being selected from acetone, 1,4-dioxane, tetrahydrofuran / isopropyl alcohol, tetrahydrofuran / acetone, tetrahydrofuran / ethyl acetate, 1,4-dioxane / isopropyl alcohol; and Step 2, filtering, washing, and drying to obtain crystalline form A.

[0029] In a preferred embodiment, the mass / volume ratio of the compound represented by formula (I) to the solvent I is 1 / (25-35) (g / mL), more preferably 1 / 30 (g / mL). When a mixed solvent such as tetrahydrofuran / isopropyl alcohol, tetrahydrofuran / acetone, tetrahydrofuran / ethyl acetate, or 1,4-dioxane / isopropyl alcohol is selected, the volume ratio of the two solvents is 1:(0.8-1.2), more preferably 1:1.

[0030] The method for preparing crystalline form B includes step 1 of dissolving the compound of formula (I) in an appropriate amount of solvent II at room temperature, leaving the compound to stand, and then volatilizing the solvent II to crystallize the compound, the solvent II being selected from tetrahydrofuran, butanone, acetonitrile / methyl tert-butyl ether, and butanone / acetone; and step 2 of filtering, washing, and drying the resulting mixture to obtain crystalline form B.

[0031] In a preferred embodiment, the mass / volume ratio of the compound represented by formula (I) to the solvent II is 1 / (25-35) (g / mL), more preferably 1 / 30 (g / mL). When a mixed solvent such as acetonitrile / methyl tert-butyl ether or butanone / acetone is selected, the volume ratio of the two solvents is 1:(0.8-1.2), more preferably 1:1.

[0032] The preparation method of crystalline form C includes the steps of: Step 1, dissolving compound of formula (I) in a suitable amount of solvent III by heating, selectively filtering the resulting clear solution while hot, and cooling the resulting solution for crystallization, said solvent III being selected from methyl isobutyl ketone; and Step 2, filtering, washing, and drying to obtain crystalline form C.

[0033] In a preferred embodiment, the mass / volume ratio of the compound represented by the formula (I) to methyl isobutyl ketone is 1 / (25 to 35) (g / mL), and more preferably 1 / 30 (g / mL).

[0034] The method for preparing crystalline form D includes the steps of: Step 1, in which the compound of formula (I) is dissolved in a suitable amount of a solvent IV by heating, followed by hot filtration, and then cooling the filtrate to crystallize, the solvent IV being selected from methanol and isopropyl alcohol; and Step 2, in which the compound of formula (I) is filtered, washed, and then dried to obtain crystalline form D.

[0035] In a preferred embodiment, the mass / volume ratio of the compound represented by the formula (I) to methanol / isopropyl alcohol is 1 / (25 to 35) (g / mL), and more preferably 1 / 30 (g / mL). In a more preferred embodiment, the volume ratio of the two solvents, methanol / isopropyl alcohol, is 1:(0.8 to 1.2), and more preferably 1:1.

[0036] The preparation method of crystalline form E includes step 1, in which compound of formula (I) is dissolved in a suitable amount of solvent V by heating, selectively filtered while hot, and the obtained clear solution is cooled to crystallize, said solvent V being selected from 1,4-dioxane / ethyl acetate; and step 2, in which the crystals are filtered, washed and dried to obtain crystalline form E.

[0037] In a preferred embodiment, the mass / volume ratio of the compound represented by the formula (I) to 1,4-dioxane / ethyl acetate is 1 / (35 to 45) (g / mL), and more preferably 1 / 40 (g / mL). In a more preferred embodiment, the volume ratio of the two solvents, 1,4-dioxane / ethyl acetate, is 1:(0.5 to 0.8), and more preferably 1:0.6.

[0038] The preparation method of crystalline form F includes the steps of: Step 1, dissolving compound of formula (I) in a suitable amount of solvent VI by heating, selectively filtering the resulting clear solution while hot, and cooling the resulting solution for crystallization, said solvent VI being selected from dichloromethane / methyl tert-butyl ether; and Step 2, filtering, washing, and drying to obtain crystalline form F.

[0039] In a preferred embodiment, the mass / volume ratio of the compound represented by the formula (I) to dichloromethane / methyl tert-butyl ether is 1 / (35 to 45) (g / mL), and more preferably 1 / 40 (g / mL).

[0040] In a more preferred embodiment, the volume ratio of the two solvents, dichloromethane / methyl tert-butyl ether, is 1:(0.8 to 1.2), and more preferably 1:1.

[0041] In a preferred embodiment, in the process of producing crystalline forms A to F, the crystals are crystallized by cooling / volatilization, then filtered, washed, and dried to obtain crystalline forms A to F. In a more preferred embodiment, the drying conditions are room temperature and reduced pressure (−0.05 to 0.5 MPa, more preferably −0.1 MPa) for 2 to 5 days (more preferably 3 days).

[0042] The preparation method of crystalline form I includes the steps of: Step 1, dissolving the compound of formula (I) in a suitable amount of solvent VII by heating, selectively filtering the resulting clear solution while hot, and cooling the resulting solution for crystallization, the solvent VII being selected from ethyl acetate and ethanol; and Step 2, filtering, washing, and drying to obtain crystalline form I.

[0043] In a preferred embodiment, the mass / volume ratio of the compound represented by formula (I) to ethyl acetate is 1 / (40-50) (g / mL), more preferably 1 / 45 (g / mL). In a preferred embodiment, the mass / volume ratio of the compound represented by formula (I) to ethanol is 1 / (10-20) (g / mL), more preferably 1 / 20 (g / mL).

[0044] In a preferred embodiment, the drying conditions are vacuum drying at 50 to 70°C for 1 to 3 days, followed by vacuum drying at 70 to 90°C for 1 to 3 days, and in a more preferred embodiment, vacuum drying at 60°C for 2 days, followed by vacuum drying at 80°C for 2 days. Unless otherwise specified, the M / N solvent referred to in the above content refers to a mixed solvent of an M solvent and an N solvent.

[0045] In the above-mentioned preparation method, if there are no impurities that require filtration after dissolving the compound represented by formula (I), i.e., if a clear solution is obtained as it is, the "hot filtration" step may be omitted, and the solution may be cooled and crystallized as it is. However, if flocs, solids, or other insoluble substances remain after dissolving the compound represented by formula (I), the "hot filtration" step is introduced.

[0046] The "compound represented by formula (I)" referred to in the above-mentioned production method may include products in various forms, for example, amorphous or any crystalline form, mixed crystalline form or any other normally existing state, unless otherwise specified. Effect of the Invention

[0047] The crystalline forms (A, B, C, D, E, F, I) of the compound represented by formula (I) obtained by the present invention have good water solubility and excellent absorbability when orally administered, and therefore can be effectively used in the treatment of malignant tumors and autoimmune diseases which are effective by inhibiting casein kinase 1ε activity. [Brief description of the drawings]

[0048] [Figure 1] 1 is a powder X-ray diffraction pattern of crystalline form A of the compound of formula (I). [Diagram 2] 1 is a powder X-ray diffraction pattern of crystalline form B of compound of formula (I). [Diagram 3] 1 is a powder X-ray diffraction pattern of crystalline form C of compound of formula (I). [Figure 4] 1 is a powder X-ray diffraction pattern of crystalline form D of compound of formula (I). [Diagram 5] 1 is a powder X-ray diffraction pattern of crystalline form E of the compound of formula (I). [Figure 6] 1 is a powder X-ray diffraction pattern of crystalline form F of compound of formula (I). [Figure 7] 1 is a powder X-ray diffraction pattern of crystalline form I of the compound of formula (I). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0049] In order to make the object and technical means of the present invention clearer, the present invention will be described in more detail below in combination with the drawings and examples of the present invention, which are only for illustrating the technical solutions of the present invention, and are not intended to limit the substance and scope of the present invention.

[0050] Test equipment used in the experiment 1. Powder X-ray diffraction spectrum Equipment: D8ADVANCE type powder X-ray diffractometer Radiation: Monochromatic Cu-Kα radiation (λ=1.5406) Scan mode: θ / 2θ, Scan range: 3~40 o 2. High performance liquid chromatography (HPLC) Equipment:Agilent1260 Chromatographic column: Shimpack VPODS150*4.6mm5μm Mobile phase: Mobile phase A: 0.1% phosphoric acid solution, Mobile phase B: methanol, eluted by isocratic elution, Mobile phase A: Mobile phase B = 20:80 Detection wavelength: 230 nm EXAMPLES

[0051] Synthesis of (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate JPEG0007603292000003.jpg114170

[0052] Step 1: Dissolve D-methyl lactate (20 g) in 200 mL of dichloromethane, cool to 0 ° C, add benzoyl chloride (27 g), DMAP (1.2 g), then add triethylamine (23.3 g) dropwise using a dropping funnel, and after the dropwise addition is completed, gradually warm to room temperature and react for 20 hours while stirring. Confirm that the reaction is complete by detection with TLC (thin layer chromatography), add 100 mL of water, stir and extract, and separate the dichloromethane phase. Add 200 mL of 0.2 M aqueous hydrochloric acid solution to the dichloromethane phase, stir, then leave to stand and separate, separate the dichloromethane layer, and concentrate under reduced pressure to obtain the crude product (R)-2-benzoyloxypropionate methyl, which is used as is in the next reaction.

[0053] Step 2: The crude product of (R)-2-benzoyloxymethyl propionate obtained in step 1 is dissolved in 250 mL of tetrahydrofuran and cooled to 0°C, and then the prepared 3% aqueous sodium hydroxide solution (250 mL) is added dropwise. During the dropping process, the temperature of the reaction system is controlled so as not to exceed 10°C, and the reaction is continued for 2 hours after the dropping is completed. After confirming that the hydrolysis reaction is complete by detection with TLC, 1.0 M aqueous hydrochloric acid is added to neutralize the solution so that the pH value is about 2 to 4, and then extracted twice with ethyl acetate (about 300 mL each time), and the ethyl acetate layers are combined and concentrated under reduced pressure to obtain a solid. Methyl tert-butyl ether (50 mL) and petroleum ether (300 mL) are added to the obtained solid, and the solid is heated to reflux for 1 hour and beaten, then gradually cooled to room temperature and kept for 12 hours, filtered and washed, and the filter cake is blown dry to obtain the compound (R)-2-(benzoyloxy)propionic acid (30.5 g). 1 HNMR(400MHz,CDCl3)δ9.89(s,1H),8.19-8.08(m,2H),7.67-7.58(m,1H),7.49(t,J=7.7Hz,2H),5.40(q,J=7.1Hz,1H),1.72(d,J=7.1Hz,3H).

[0054] Step 3: 2-Amino-6-chlorobenzoic acid (10.0 g) and (R)-2-(benzoyloxy)propionic acid (13.6 g) are dissolved in 100 mL of DMF, pyridine (50.7 g) is added, and the mixture is heated to 50° C. with stirring. Triphenyl phosphite (21.7 g) is then gradually added and reacted for 12 hours, after which aniline (8.2 g) and triphenyl phosphite (10.8 g) are added and the mixture is reacted for about 12 hours with continuous stirring at 50° C. After confirming the completion of the reaction by TLC detection, cool to room temperature, extract with ethyl acetate and 1M aqueous hydrochloric acid, separate the liquids, wash the ethyl acetate phase again with 1M aqueous hydrochloric acid, collect the ethyl acetate layer, and concentrate under reduced pressure to obtain an oily substance, then add a mixture of methyl tert-butyl ether and cyclohexane (1:5), stir and beat, and filter to obtain the solid product (R)-1-(5-chloro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl benzoate (16.1g). 1 HNMR(500MHz,DMSO)δ8.01-7.88(m,2H),7.77(t,J=8.0Hz,1H),7.71-7.63(m,2H),7.60(dd,J=10.3,2.6Hz,3 H),7.51(dd,J=13.7,5.8Hz,3H),7.38(dt,J=15.1,7.7Hz,2H),5.36(q,J=6.4Hz,1H),1.55(d,J=6.5Hz,3H).

[0055] Step 4: (R)-1-(5-chloro-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)ethyl benzoate (10.2 g) is dissolved in 100 mL of methanol, lithium hydroxide (0.9 g) is added, and the mixture is reacted at room temperature with stirring for about 4 hours. The reaction is confirmed to be complete by TLC detection, and dichloromethane and water are added for extraction. The dichloromethane layer is separated and concentrated under reduced pressure to obtain a crude product. The crude product is refluxed with methyl tert-butyl ether and triturated, filtered, and dried to obtain the compound (R)-5-chloro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one (7.0 g). Chiral purity ee%: 98.7%. Chiral-HPLC is measured on a reverse-phase CHIRALPAKIG-3 column, and the retention time is 52.7 min. 1 HNMR(500MHz,CDCl3)δ7.70-7.62(m,2H),7.60-7.52(m,3H),7.52-7.49(m,1H),7.32( dd,J=7.5,2.2Hz,1H),7.27-7.23(m,1H),4.45(q,J=6.4Hz,1H),1.24(d,J=6.4Hz,3H).

[0056] Step 5: (R)-5-chloro-2-(1-hydroxyethyl)-3-phenylquinazolin-4(3H)-one (6.0g) is dissolved in 75mL of NMP, 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine (5.8g), triphenylphosphine (7.8g) and DIAD (6.0g) are added, and the mixture is reacted at room temperature for about 12 hours with stirring. After the reaction is confirmed to be complete by TLC detection, the reaction liquid is added directly to water to precipitate a solid, and the filter cake is collected by suction filtration. The filter cake is refluxed with acetonitrile, beaten, filtered, and the filter cake is blown dry to obtain the solid product (S)-2-(1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (5.9g). 1HNMR(500MHz,DMSO)δ7.88(s,1H),7.83(t,J=8.0Hz,1H),7.74(dd,J=8.2,1.1Hz,1H),7.64(dd,J=7.9,1.1Hz,1H),7.58(d,J=8.1Hz,1H),7.4 3(td,J=7.7,1.3Hz,1H),7.14(t,J=7.5Hz,1H),6.88(td,J=7.8,1.3Hz,1H),6.33(d,J=7.9Hz,1H),5.86-5.72(m,1H),1.68(d,J=6.7Hz,3H).

[0057] Step 6: (S)-2-(1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (5.0 g) and (2,3-difluoro-4-methoxyphenyl)boronic acid (2.6 g) were dissolved in a mixed solvent of 20 mL of 2-methyltetrahydrofuran and 5 mL of water, and the solution was added with [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (330 mg) and sodium carbonate. Add 2.4g of ethyl acetate and heat to 80℃ for about 6 hours. After confirming the completion of the reaction by TLC, concentrate under reduced pressure. Purify the obtained residue by silica column chromatography to obtain white solid product (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazole[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (4.1g). Chiral purity ee%: 98.6%. Chiral-HPLC is measured on a normal phase CHIRALCELOD-H column, retention time is 12.09min. 1HNMR(500MHz,CDCl3)δ8.05(s,1H),7.73(d,J=8.1Hz,1H),7.64(t,J=8.0Hz,1H),7.51 (d,J=7.8Hz,1H),7.47(t,J=7.6Hz,1H),7.29(d,J=7.9Hz,1H),7.22(dd,J=11.8,4.4H z,1H),7.16(t,J=7.4Hz,1H),6.93-6.84(m,2H),6.53(d,J=8.0Hz,1H),6.00(q,J=6.7 Hz,1H),5.33(brs,2H),3.96(s,3H),1.90(d,J=6.7Hz,3H).LC-MS(ESI-MS):560[M+H] + . EXAMPLES

[0058] Preparation of (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate

[0059] The (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one (3.4 g) synthesized in Example 1 is added to a reaction flask, then 50 mL of isopropyl alcohol and methanesulfonic acid (590 mg) are added, and the mixture is heated to reflux for about 1 hour until the solid is completely dissolved and becomes a clear solution. Then, the mixture is slowly cooled to room temperature, desolvated, and filtered to collect the solid. The solid obtained is vacuum dried at room temperature to obtain the white solid product (S)-2-(1-(4-amino-3-(2,3-difluoro-4-methoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-5-chloro-3-phenylquinazolin-4(3H)-one-methanesulfonate (3.65 g). 1HNMR(500MHz,DMSO)δ8.14(s,1H),7.84(t,J=8.0Hz,1H),7.75(d,J=8.1Hz,1H),7.65(t,J=7.7Hz,2H),7.47(t,J=7.6Hz,1H),7.31(t,J=8.1Hz,1H) ,7.15(dt,J=16.3,7.8Hz,2H),6.92(t,J=7.5Hz,1H),6.41(d,J=7.8Hz,1H ),5.97(t,J=6.6Hz,1H),3.93(s,3H),2.36(s,3H),1.77(d,J=6.6Hz,3H). LC-MS(ESI-MS):560[M+H] + . EXAMPLES

[0060] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, and then 30mL of acetone was added. The solution was heated to a clear solution, and then cooled to crystallize, filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 60%. The powder X-ray diffraction analysis showed that the crystal form was A. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 7.848, 11.618, 15.562, 15.835, 20.185, 24.799, 25.655 and deviations of ±0.2°. The results are shown in Table 1 and Figure 1.

[0061] Table 1. Powder X-ray diffraction data for crystalline form A JPEG0007603292000004.jpg255159 EXAMPLES

[0062] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of 1,4-dioxane was added. The solution was heated to a clear solution, and then cooled to cause crystallization. The crystal was filtered, washed, and then dried (under reduced pressure at room temperature for 3 days, -0.1 MPa, the same applies below). The yield was 68%, and the crystal form was determined to be A by powder X-ray diffraction. EXAMPLES

[0063] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of tetrahydrofuran / isopropyl alcohol (volume ratio 1:1) was added. The solution was heated to become clear, and then cooled to cause crystallization. The crystal was filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 54%, and the crystal form was determined to be Form A by powder X-ray diffraction. EXAMPLES

[0064] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of tetrahydrofuran / acetone (volume ratio 1:1) was added and heated to obtain a clear solution. The solution was then cooled to cause crystallization, filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 50%, and the crystal form was determined to be A by powder X-ray diffraction. EXAMPLES

[0065] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of tetrahydrofuran / ethyl acetate (volume ratio 1:1) was added. The solution was heated to become clear, and then cooled to cause crystallization. The crystal was filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 62%, and the crystal form was determined to be A by powder X-ray diffraction. EXAMPLES

[0066] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of 1,4-dioxane / isopropyl alcohol (volume ratio 1:1) was added and heated to obtain a clear solution. The solution was then cooled to cause crystallization, filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 54%, and the crystal form was determined to be Form A by powder X-ray diffraction. EXAMPLES

[0067] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of tetrahydrofuran was added. The solution was stirred at room temperature to obtain a clear solution, and then the solution was allowed to stand at room temperature for 2 days to allow evaporation and crystallization. The crystals were filtered, washed, and then dried (under reduced pressure at room temperature for 3 days), and the result of powder X-ray diffraction analysis was that the crystal form was B. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 10.739, 11.961, 13.726, 14.374, 23.714, 24.057, 25.033, and deviations of ±0.2°, and the results are shown in Table 2 and Figure 2.

[0068] Table 2. Powder X-ray diffraction data for crystalline form B JPEG0007603292000005.jpg250161 EXAMPLES

[0069] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of butanone was added. The solution was stirred at room temperature to obtain a clear solution, and then the solution was allowed to stand at room temperature for 2 days to allow evaporation and crystallization. The crystals were filtered, washed, and then dried (dried under reduced pressure at room temperature for 3 days). The yield was 82%, and the crystal form was determined to be crystalline form B by powder X-ray diffraction. EXAMPLES

[0070] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of acetonitrile / methyl tert-butyl ether (volume ratio 1:1) was added and stirred at room temperature to obtain a clear solution. The solution was then allowed to stand at room temperature for 2 days to evaporate and crystallize. The crystals were filtered, washed, and then dried (dried under reduced pressure at room temperature for 3 days). The yield was 54%, and the crystal form was determined to be form B by powder X-ray diffraction. EXAMPLES

[0071] 1 g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30 mL of a mixed solvent of butanone / acetone (volume ratio 1:1) was added and stirred at room temperature to obtain a clear solution. The solution was then allowed to stand at room temperature for 2 days to evaporate and crystallize. The crystals were filtered, washed, and then dried (dried under reduced pressure at room temperature for 3 days). The yield was 48%, and the crystal form was determined to be crystalline form B by powder X-ray diffraction. EXAMPLES

[0072] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, and then 30mL of methyl isobutyl ketone was added. The solution was heated to a clear solution, and then cooled to crystallize, filtered, washed, and then dried (under reduced pressure at room temperature for 3 days). The yield was 80%, and the result of powder X-ray diffraction showed that the crystal form was C. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 7.319, 8.092, 11.763, 14.728, 15.855, 16.265 and deviations of ±0.2°, and the results are shown in Table 3 and Figure 3.

[0073] Table 3. Powder X-ray diffraction data for crystalline form C JPEG0007603292000006.jpg221170 EXAMPLES

[0074] 1g of the compound represented by formula (I) (prepared in Example 2) was placed in a glass bottle, and then 30mL of methanol / isopropyl alcohol (1:1) was added. The solution was heated to a clear solution, and then cooled to crystallize, filtered, washed, and dried (under reduced pressure at room temperature for 3 days). The yield was 58%, and the powder X-ray diffraction analysis showed that the crystal form was D. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 11.308, 17.237, 18.568, 20.213, 21.148, and 21.293 and deviations of ±0.2°, and the results are shown in Table 4 and Figure 4.

[0075] Table 4. Powder X-ray diffraction data for crystalline form D JPEG0007603292000007.jpg255168 EXAMPLES

[0076] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, and then 40mL of a mixed solvent of 1,4-dioxane / ethyl acetate (volume ratio 5:3) was added, and the solution was heated to become clear, and then cooled to crystallize, filtered, washed, and then dried (dried under reduced pressure at room temperature for 3 days). The yield was 64%, and the result of powder X-ray diffraction was that the crystal form was E. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 7.114, 7.936, 9.973, 11.040, 15.504, 16.507, 20.071, 21.555 and deviations of ±0.2°, and the measurement results are shown in Table 5 and Figure 5.

[0077] Table 5. Powder X-ray diffraction data for crystalline form E JPEG0007603292000008.jpg255147 EXAMPLES

[0078] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, and then 40mL of a mixed solvent of dichloromethane / methyl tert-butyl ether (volume ratio = 1:1) was added, and the solution was heated to become clear, and then cooled to crystallize. The crystals were filtered, washed, and dried (dried under reduced pressure at room temperature for 3 days), with a yield of 82%. The powder X-ray diffraction analysis revealed that the crystal was of crystalline form F. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 9.959, 13.604, 14.830, 20.645, 23.717, and 26.262, with deviations of ±0.2°, and the results are shown in Table 6 and Figure 6.

[0079] Table 6. Powder X-ray diffraction data for crystalline form F JPEG0007603292000009.jpg162170 EXAMPLES

[0080] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, and then 40mL of ethanol was added. The solution was heated to a clear solution, and then cooled to cause crystallization. The crystal was filtered, washed, and then dried (vacuum dried at 60°C for 2 days, and vacuum dried at 80°C for 2 days). The yield was 50%, and the crystal form was determined to be Form I by powder X-ray diffraction. EXAMPLES

[0081] 1g of the compound represented by formula (I) (prepared according to Example 2) was placed in a glass bottle, then 45mL of ethyl acetate was added, and the solution was heated to a clear solution, followed by cooling to crystallize, filtering, washing, and drying (vacuum drying at 60°C for 2 days, and vacuum drying at 80°C for 2 days), with a yield of 66%. Powder X-ray diffraction analysis revealed that the crystal was of crystalline form I. The main characteristic peaks were characteristic powder X-ray diffraction peaks at 2θ values ​​of 7.321, 7.538, 12.317, 14.643, 15.839, and 19.458, with deviations of ±0.2°, and the results are shown in Table 7 and Figure 7.

[0082] Table 7. Powder X-ray diffraction data for crystalline form I JPEG0007603292000010.jpg131164 EXAMPLES

[0083] Solubility measurement of crystal forms A, B, C, D, E, and F

[0084] Preparation of standard curve: A series of compounds are precisely weighed and placed in a measuring flask, and then a suitable amount of acetonitrile is added to dissolve and dilute to the scale to obtain a stock solution. A series of high-concentration standard solutions and low-concentration standard solutions are precisely weighed and placed in a measuring flask, and then mobile phase is added to dilute to the scale, and then shaken to obtain a homogeneous solution. Load each solution in order from low to high concentration, and record the peak area. A linear regression is performed with the concentration of the compound as the abscissa and the peak area as the ordinate to obtain the linear regression equation of the compound, that is, obtain the standard curve of the solubility of the compound represented by formula (I). Measurement of equilibrium solubility: Put an excess amount of compound into a 10mL EP tube with a cap, then add 2mL of purified water, and at room temperature, treat with ultrasonic waves for 15 minutes, then leave to stand for 20 minutes. This operation is repeated 4 times, then centrifuge (3000r / min, 10min) and take the supernatant. The supernatant is appropriately diluted with a solvent, then filtered through a microfiltration membrane, and the filtrate is measured by HPLC, the peak area is recorded, and the saturated solubility of each compound in water is calculated by substituting it into the standard curve. The measurement results are shown in Table 8.

[0085] Table 8. Solubility results in water for crystal forms A, B, C, D, E, and F JPEG0007603292000011.jpg85170The results revealed that crystalline forms A, B, C, D, E, and F of the compound represented by formula (I) were all soluble in water. EXAMPLES

[0086] In vivo pharmacokinetic study in rats

[0087] Experimental method: SD rats were used as experimental animals, and 10 mg / kg (prepared into a milky suspension with sodium carboxymethylcellulose solution) was administered intragastrically. Blood was collected at 0.17, 0.33, 0.5, 1, 1.5, 2, 4, 6, 8, 12, and 24 hours after intragastric administration. 0.3 mL of whole blood was collected, and 0.1 mL of plasma was collected after centrifugation and analyzed by LC-MS.

[0088] Table 9. Blood concentration measurement results after oral administration in SD rats JPEG0007603292000012.jpg50170

[0089] The pharmacokinetic properties of the crystalline forms C, D and F of the compound of formula (I) in vivo in rats were examined with reference to the free base of the compound of formula (I). The measurement results revealed that the pharmacokinetic properties of the compound of formula (I) were significantly improved compared to the free base. Therefore, the compound of the present invention can be used for the treatment of related diseases by oral administration. EXAMPLES

[0090] In vivo pharmacokinetic study in dogs

[0091] Experimental method: Beagles were used as experimental animals, and 10 mg / kg (prepared into a milky suspension with sodium carboxymethylcellulose solution) was administered intragastrically. Blood was collected at 0, 0.5, 1, 2, 3, 4, 6, 8, and 24 hours after intragastric administration. 0.3 mL of whole blood was collected, and 0.1 mL of plasma was collected after centrifugation and analyzed by LC-MS.

[0092] Table 10. Blood concentration measurement results after oral administration in dogs JPEG0007603292000013.jpg41170In Table 10, NA indicates below the lower limit of detection. AUC: area under the blood concentration-time curve.

[0093] The pharmacokinetic properties of the crystalline form F of the compound of formula (I) in vivo in beagle dogs were examined with reference to the free base of the compound of formula (I). The measurement results revealed that the pharmacokinetic properties of the compound of formula (I) were significantly improved compared to the free base. Therefore, the compound of the present invention can be used for the treatment of related diseases by oral administration.

Claims

1. A crystalline form A of the casein kinase 1 ε inhibitor represented by formula (I), The powder X-ray diffraction pattern of the crystalline form A has characteristic peaks at 2θ values ​​of 7.848±0.2°, 11.618±0.2°, 15.562±0.2°, 15.853±0.2°, 20.185±0.2°, and 25.655±0.2°.

2. The crystalline form A of the casein kinase 1 ε inhibitor according to claim 1, characterized in that it simultaneously has characteristic peaks at 2θ values ​​of 11.926±0.2°, 15.271±0.2°, 21.851±0.2° and 24.799±0.2°.

3. The method for preparing crystalline form A according to claim 1 or 2, comprising the steps of heating the compound of formula (I) to dissolve it in a solvent I, selectively filtering the solution while hot, and cooling the resulting clear solution to crystallize the compound to obtain crystalline form A, wherein the solvent I is selected from acetone, 1,4-dioxane, tetrahydrofuran / isopropyl alcohol, tetrahydrofuran / acetone, tetrahydrofuran / ethyl acetate, and 1,4-dioxane / isopropyl alcohol.

4. Crystalline form B of the compound of formula (I), The powder X-ray diffraction pattern of crystalline form B is characterized by having characteristic peaks at 2θ values ​​of 10.739±0.2°, 11.961±0.2°, 13.726±0.2°, 14.374±0.2°, 23.714±0.2°, 24.057±0.2°, and 25.033±0.2°.

5. The crystalline form B according to claim 4, characterized in that it simultaneously has characteristic peaks at 2θ values ​​of 14.809±0.2°, 18.574±0.2°, 22.313±0.2°, 26.125±0.2° and 26.659±0.2°.

6. 6. The method for preparing crystalline form B according to claim 4 or 5, comprising the steps of dissolving the compound of formula (I) in a solvent II at room temperature, allowing the compound to stand and then volatilizing the solvent to crystallize the compound to obtain crystalline form B, wherein the solvent II is selected from the group consisting of tetrahydrofuran, butanone, acetonitrile / methyl tert-butyl ether, and butanone / acetone.

7. Crystalline form C of the compound of formula (I), The powder X-ray diffraction pattern of crystalline form C has characteristic peaks at 2θ values ​​of 7.319±0.2°, 8.092±0.2°, 11.763±0.2°, 14.728±0.2°, 15.855±0.2°, and 16.265±0.2°.

8. 8. The method for preparing crystalline form C according to claim 7, comprising the steps of heating the compound of formula (I) to dissolve it in a solvent III, selectively hot filtering the resulting clear solution, and cooling the resulting solution to crystallize to obtain crystalline form C, wherein the solvent III is selected from methyl isobutyl ketone.

9. Crystal form D of the compound of formula (I), The powder X-ray diffraction pattern of the crystalline form D is characterized by having characteristic peaks at 2θ values ​​of 11.308±0.2°, 17.237±0.2°, 18.568±0.2°, 20.213±0.2°, 21.148±0.2°, and 21.293±0.2°.

10. 10. The crystalline form D according to claim 9, characterized in that it has characteristic peaks at 2θ values ​​of 10.341±0.2°, 13.819±0.2°, 17.982±0.2°, 20.804±0.2°, 22.729±0.2°, 25.035±0.2°, 25.289±0.2° and 28.008±0.2°.

11. The method for preparing crystalline form D according to claim 9 or 10, comprising the steps of heating the compound of formula (I) to dissolve it in a solvent IV, selectively hot filtering the resulting clear solution, and cooling the solution to crystallize the resulting crystalline form D, wherein the solvent IV is selected from methanol / isopropyl alcohol.

12. Crystalline form E of compound of formula (I), The powder X-ray diffraction pattern of crystalline form E is characterized by having characteristic peaks at 2θ values ​​of 7.114±0.2°, 7.936±0.2°, 9.973±0.2°, 11.040±0.2°, 14.484±0.2°, 15.504±0.2°, 16.507±0.2°, 20.071±0.2°, and 21.555±0.

2.

13. The crystalline form E according to claim 12, characterized in that it simultaneously has characteristic peaks at 2θ values ​​of 5.492±0.2°, 22.519±0.2°, 24.464±0.2°, and 27.980±0.2°.

14. The method for preparing crystalline form E according to claim 12 or 13, comprising the steps of heating and dissolving compound of formula (I) in a solvent V, selectively hot filtering, and cooling the resulting clear solution to crystallize to obtain crystalline form E, wherein the solvent V is selected from 1,4-dioxane / ethyl acetate.

15. Crystalline form F of the compound of formula (I), The powder X-ray diffraction pattern of crystalline form F is characterized by having characteristic peaks at 2θ values ​​of 9.959±0.2°, 13.604±0.2°, 14.830±0.2°, 20.645±0.2°, 23.717±0.2°, and 26.262±0.2°.

16. 16. The method for preparing crystalline form F according to claim 15, comprising the steps of heating compound of formula (I) to dissolve in a suitable amount of solvent VI, selectively hot filtering, and cooling the resulting clear solution to crystallize to obtain crystalline form F, wherein the solvent VI is selected from dichloromethane / methyl tert-butyl ether.

17. 20. Use of the crystalline form of any one of claims 1, 2, 4, 5, 7, 9, 10, 12, 13, 15 in the manufacture of a drug for treating malignant tumors and autoimmune diseases which are mediated by inhibiting casein kinase 1ε activity.

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