Salts of a degradation btk compound and crystalline forms thereof and medical uses thereof

Abstract

Provided are a salt and / or a crystal form of a compound degrading BTK and preparation and application thereof. The pharmaceutically acceptable salt and crystal form of the compound as shown in formula (I), wherein the pharmaceutically acceptable salt is selected from a maleate salt, a fumarate salt, a hydrogen halide salt (preferably a hydrobromide salt and a hydrochloride salt), a sulfate salt, a phosphate salt, an L-tartrate salt, a citrate salt, an L-malate salt, a hippurate salt, a D-glucuronate salt, a glycolate salt, a mucate salt, a succinate salt, a lactate salt, an orotate salt, a pamoate salt, a glycine salt, an alanine salt, an arginine salt, a cinnamate salt, a benzoate salt, a benzene sulfonate salt, a p-toluene sulfonate salt, an acetate salt, a propionate salt, a valerate salt, a triphenylacetic acid salt, an L-proline salt, a ferulic acid salt, a 2-hydroxyethanesulfonate salt, a mandelate salt, a nitrate salt, a methanesulfonate salt, a malonate salt, a gentisate salt, a salicylate salt, an oxalate salt or a glutarate salt:

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more specifically, to the crystal form, preparation, and application of a salt that degrades BTK compounds. Background Technology

[0002] Bruton's tyrosine kinase (BTK) is a member of the Tec family of non-receptor protein tyrosine kinases and a key regulator in the B-cell antigen receptor (BCR) signaling pathway, distributed in the lymphatic, hematopoietic, and blood systems. BTK mutations activate downstream tumor cell proliferation, differentiation, and angiogenesis signaling pathways, leading to X-linked agammaglobulinemia, non-Hodgkin's lymphoma (NHL), and many B-cell malignancies, including chronic lymphocytic leukemia (CLL), mantle cell lymphoma, and diffuse large B-cell lymphoma. Because it is primarily expressed in B cells and myeloid cells, BTK is a target with good targeting and safety profile.

[0003] PROTAC (proteolysis targeting chimera) molecules are bifunctional compounds that can simultaneously bind to target proteins and E3 ubiquitin ligases. These compounds induce the target protein to be recognized by the cell's proteasome, causing its degradation and effectively reducing its concentration in cells. By introducing ligands that bind to different target proteins into PROTAC molecules, the application of PROTAC technology in the treatment of various diseases has become possible, and this technology has received widespread attention in recent years. Summary of the Invention

[0004] The purpose of this invention is to provide a novel, highly effective BTK-degrading compound, its pharmaceutical composition, and its use in the antitumor field. The BTK-degrading compound of this invention exhibits good stability (including chemical and crystal form stability), ease of oral administration, and good solubility and bioavailability.

[0005] The purpose of this invention is to provide a novel, pharmaceutically acceptable salt of a BTK-degrading compound or crystals of a BTK-degrading compound and its pharmaceutically acceptable salt, pharmaceutical compositions thereof, and their use in the field of antitumor therapy.

[0006] The crystals of this invention are easy to process and crystallize, have good stability, are easy to take orally, and have good solubility and bioavailability.

[0007] Another object of the present invention is to provide a method for preparing the compound and / or crystals of the BTK degradation.

[0008] Another object of the present invention is to provide a pharmaceutical composition containing the degraded BTK compound and / or crystals thereof.

[0009] Another object of the present invention is to provide the application of the compounds and / or crystals that degrade BTK.

[0010] This invention provides a pharmaceutically acceptable salt of the compound shown in formula (I).

[0011]

[0012] In some embodiments, Cy1 or Cy2 is independently selected from piperidinyl or aziridine.

[0013] In some implementations, Cy1 or Cy2 is selected independently.

[0014] In some embodiments, the pharmaceutically acceptable salt of the compound represented by formula (I) is selected from maleate, fumarate, hydrohalate (preferably hydrobromide and hydrochloride), sulfate, phosphate, L-tartrate, citrate, L-malate, hippurate, D-glucuronide, glycolate, mucilage, succinate, lactate, orotate, pamoate, glycinate, alanine, arginine, cinnamate, benzoate, benzenesulfonate, p-toluenesulfonate, acetate, propionate, valerate, triphenylacetate, L-proline, ferulic acid, 2-hydroxyethanesulfonate, mandelate, nitrate, methanesulfonate, malonate, gentianate, salicylate, oxalate, or glutarate.

[0015] In some embodiments, the hydrohalate is a hydrobromide or hydrochloride.

[0016] In some embodiments, the molar ratio of the compound (free base) represented by formula (I) to different acids is about 1:1, 1:1.5, 1:2, 1:2.5 or 1:3.

[0017] The present invention also provides pharmaceutically acceptable salts of compounds represented by formula (Ia) or (Ib) below.

[0018]

[0019] In some embodiments, the pharmaceutically acceptable salt of the compound shown in (Ia) or (Ib) is selected from maleate, fumarate, hydrohalate (preferably hydrobromide and hydrochloride), sulfate, phosphate, L-tartrate, citrate, L-malate, hippurate, D-glucuronide, glycolate, mucilage, succinate, lactate, orotate, pamoate, glycinate, alanine, arginine, cinnamate, benzoate, benzenesulfonate, p-toluenesulfonate, acetate, propionate, valerate, triphenylacetate, L-proline, ferulic acid, 2-hydroxyethanesulfonate, mandelate, nitrate, methanesulfonate, malonate, gentianate, salicylate, oxalate, or glutarate, preferably maleate, fumarate, L-tartrate, citrate, L-malate, salicylate, or oxalate.

[0020] In some embodiments, the pharmaceutically acceptable salt of the compound represented by formula (Ia) is selected from maleate salts, and the molar ratio of the compound represented by formula (Ia) to the maleate salt is about 1:1, 1:1.5, 1:2, 1:2.5 or 1:3.

[0021] In some embodiments, the pharmaceutically acceptable salt of the compound shown in formula (I) has the structure shown in formula (II).

[0022] The present invention also provides compounds represented by the following formula (II),

[0023]

[0024] The present invention also provides crystal form I of the compound shown in formula (II), which, when irradiated with Cu-Kα, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 5.96°±0.2°, 9.30°±0.2°, 11.86°±0.2°, 15.80°±0.2°, 21.75°±0.2° and 23.93°±0.2°.

[0025] Preferably, the compound represented by formula (II) of the present invention is crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 3.98°±0.2°, 7.65°±0.2°, 10.87°±0.2°, 16.88°±0.2°, 17.89°±0.2°, and 26.21°±0.2°.

[0026] More preferably, the compound represented by formula (II) of the present invention is crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 15.29°±0.2°, 17.33°±0.2°, 18.55°±0.2°, 19.21°±0.2°, 19.91°±0.2°, and 22.41°±0.2°.

[0027] More preferably, the compound represented by formula (II) of the present invention is crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 4.72°±0.2°, 9.58°±0.2°, 9.92°±0.2°, 12.85°±0.2°, 13.37°±0.2°, 13.75°±0.2°, 14.45°±0.2°, 27.37°±0.2°, 28.43°±0.2°, 30.27°±0.2°, 31.51°±0.2°, and 34.21°±0.2°.

[0028] In some embodiments, the compound represented by formula (II) of the present invention is crystal form I, and its X-ray powder diffraction pattern is basically as shown in Figure 28.

[0029] In some embodiments, the compound represented by formula (II) of the present invention is crystal form I, and its differential scanning calorimetry (DSC) curve is shown in Figure 29 or its thermogravimetric analysis curve is shown in Figure 30.

[0030] The present invention also provides an amorphous form of the compound shown in formula (II), whose X-ray powder diffraction pattern using Cu-Kα radiation is basically as shown in Figure 31.

[0031] In some embodiments, the amorphous form of the compound represented by formula (II) of the present invention has a differential scanning calorimetry (DSC) curve as shown in Figure 32 or a thermogravimetric analysis (TGA) curve as shown in Figure 33.

[0032] The present invention also provides crystal form II of the compound shown in formula (II), which, when irradiated with Cu-Kα, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 3.98°±0.2°, 6.35°±0.2°, 8.10°±0.2°, 9.66°±0.2°, 12.21°±0.2°, 15.79°±0.2°, 16.75°±0.2°, and 19.39°±0.2°.

[0033] Preferably, the compound represented by formula (II) of the present invention is crystal form II, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 12.78°±0.2°, 16.33°±0.2°, 17.13°±0.2°, 17.41°±0.2°, 20.45°±0.2°, 21.43°±0.2°, 23.23°±0.2°, 24.65°±0.2°, and 25.75°±0.2°.

[0034] In some embodiments, the compound represented by formula (II) of the present invention is crystal form II, and its X-ray powder diffraction pattern is basically as shown in Figure 34.

[0035] In some embodiments, the compound represented by formula (II) of the present invention is crystal form II, and its differential scanning calorimetry (DSC) curve is shown in Figure 35 or its thermogravimetric analysis curve is shown in Figure 36.

[0036] The present invention also provides an amorphous form of the compound shown in formula (Ia), whose X-ray powder diffraction pattern using Cu-Kα radiation is basically as shown in Figure 1.

[0037] In some embodiments, the amorphous form of the compound represented by formula (Ia) of the present invention has a differential scanning calorimetry (DSC) curve as shown in Figure 2 or a thermogravimetric analysis (TGA) curve as shown in Figure 3.

[0038] The present invention also provides that the compound represented by formula (Ia) is of crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ positions: 8.32°±0.2°, 15.69°±0.2°, 16.41°±0.2°, 17.57°±0.2°, 18.89°±0.2°, and 19.75°±0.2°.

[0039] Preferably, the compound represented by formula (Ia) of the present invention is crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 10.94°±0.2°, 11.90°±0.2°, 13.30°±0.2°, 14.39°±0.2°, 16.67°±0.2°, 17.24°±0.2°, 18.00°±0.2°, 21.25°±0.2°, 22.27°±0.2°, 23.85°±0.2°, and 26.45°±0.2°.

[0040] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form I, and its X-ray powder diffraction pattern is basically as shown in Figure 4.

[0041] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form I, and its differential scanning calorimetry (DSC) curve is shown in Figure 5 or its thermogravimetric analysis curve is shown in Figure 6.

[0042] The present invention also provides crystal form II of the compound shown in formula (Ia), which, when irradiated with Cu-Kα, has X-ray powder diffraction patterns with characteristic diffraction peaks at the following 2θ positions: 4.98°±0.2°, 7.86°±0.2°, 13.72°±0.2°, 17.65°±0.2° and 20.01°±0.2°.

[0043] Preferably, the compound represented by formula (Ia) of the present invention is crystal form II, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, further exhibits characteristic diffraction peaks at the following 2θ positions: 5.48°±0.2°, 13.43°±0.2°, 14.93°±0.2°, 15.90°±0.2°, 16.57°±0.2°, 16.95°±0.2°, 21.29°±0.2°, 22.05°±0.2°, 24.97°±0.2°, and 25.77°±0.2°.

[0044] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form II, and its X-ray powder diffraction pattern is basically as shown in Figure 7.

[0045] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form II, and its differential scanning calorimetry (DSC) curve is shown in Figure 8 or its thermogravimetric analysis curve is shown in Figure 9.

[0046] The present invention also provides crystal form III of the compound shown in formula (Ia), which, when irradiated with Cu-Kα, exhibits characteristic diffraction peaks at the following 2θ positions in its X-ray powder diffraction pattern: 5.02°±0.2°, 8.04°±0.2°, 16.91°±0.2°, 17.23°±0.2°, 18.19°±0.2°, 19.41°±0.2°, and 20.03°±0.2°.

[0047] Preferably, the crystal form III of the compound shown in formula (Ia), when irradiated with Cu-Kα, has an X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 12.36°±0.2°, 14.60°±0.2°, 15.03°±0.2°, 15.73°±0.2°, 20.57°±0.2°, 21.31°±0.2°, and 25.45°±0.2°.

[0048] More preferably, the crystal form III of the compound shown in formula (Ia), when irradiated with Cu-Kα, has an X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 5.19°±0.2°, 16.32°±0.2°, 18.75°±0.2°, 19.73°±0.2°, 21.91°±0.2°, 22.41°±0.2°, 23.48°±0.2°, 23.95°±0.2°, and 26.33°±0.2°.

[0049] More preferably, the crystal form III of the compound shown in formula (Ia), when irradiated with Cu-Kα, has an X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 10.34°±0.2°, 24.85°±0.2°, 26.93°±0.2°, 27.57°±0.2°, 28.41°±0.2°, 29.59°±0.2°, 30.19°±0.2°, 31.77°±0.2°, 33.13°±0.2°, and 35.75°±0.2°.

[0050] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form III, and its X-ray powder diffraction pattern is basically as shown in Figure 10.

[0051] In some embodiments, the compound represented by formula (Ia) of the present invention is crystal form III, and its differential scanning calorimetry (DSC) curve is shown in Figure 11 or its thermogravimetric analysis curve is shown in Figure 12.

[0052] The present invention also provides that the compound represented by formula (Ib) is of crystal form I, and its X-ray powder diffraction pattern, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ positions: 4.38°±0.2°, 8.66°±0.2°, 13.06°±0.2°, 14.34°±0.2°, 18.18°±0.2°, 20.28°±0.2°, and 21.82°±0.2°.

[0053] Preferably, the crystal form I of the compound shown in formula (Ib), when irradiated with Cu-Kα, has an X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 11.92°±0.2°, 12.74°±0.2°, and 17.44°±0.2°.

[0054] More preferably, the compound of formula (Ib) in crystal form I, when irradiated with Cu-Kα, exhibits a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 9.76°±0.2°, 11.26°±0.2°, 14.14°±0.2°, 17.04°±0.2°, 23.23°±0.2°, 24.06±0.2°, 25.26°±0.2°, and 26.42±0.2°. In some embodiments, the compound of formula (Ib) of the present invention is in crystal form I, and its X-ray powder diffraction pattern is substantially as shown in Figures 13-1 and / or 13-2.

[0055] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form I, and its differential scanning calorimetry (DSC) curve is shown in Figure 14 or its thermogravimetric analysis curve is shown in Figure 15.

[0056] The present invention also provides crystal form II of the compound shown in formula (Ib), which, when irradiated with Cu-Kα, has X-ray powder diffraction patterns with characteristic diffraction peaks at the following 2θ positions: 5.12°±0.2°, 6.68°±0.2°, 16.50°±0.2° and 20.18°±0.2°.

[0057] Preferably, the crystal form II of the compound of formula (Ib) of the present invention, when subjected to Cu-Kα radiation, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 9.98°±0.2°, 13.44°±0.2°, 13.86°±0.2°, 15.34°±0.2°, 22.40°±0.2°, and 23.12°±0.2°.

[0058] More preferably, the crystal form II of the compound of formula (Ib) of the present invention, when subjected to Cu-Kα radiation, has a X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 15.76°±0.2°, 20.99°±0.2°, 24.14°±0.2° and 26.28°±0.2°.

[0059] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form II, and its X-ray powder diffraction pattern is substantially as shown in Figures 16-1 and / or 16-2.

[0060] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form II, and its differential scanning calorimetry (DSC) curve is shown in Figure 17 or its thermogravimetric analysis curve is shown in Figure 18.

[0061] The present invention also provides crystal form III of the compound shown in formula (Ib), which, when irradiated with Cu-Kα, has X-ray powder diffraction patterns with characteristic diffraction peaks at the following 2θ positions: 7.48°±0.2°, 12.24°±0.2°, 20.50°±0.2° and 25.77°±0.2°.

[0062] Preferably, the crystal form III of the compound of formula (Ib) of the present invention, when subjected to Cu-Kα radiation, has a X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 15.59°±0.2°, 18.74°±0.2°, and 23.85°±0.2°.

[0063] More preferably, the crystal form III of the compound of formula (Ib) of the present invention, when subjected to Cu-Kα radiation, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 14.95°±0.2°, 16.18°±0.2°, 16.70°±0.2°, 19.00°±0.2° and 21.39°±0.2°.

[0064] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form III, and its X-ray powder diffraction pattern is substantially as shown in Figures 19-1 and / or 19-2.

[0065] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form III, and its differential scanning calorimetry (DSC) curve is shown in Figure 20 or its thermogravimetric analysis curve is shown in Figure 21.

[0066] The present invention also provides crystal form IV of the compound shown in formula (Ib), which, when irradiated with Cu-Kα, has X-ray powder diffraction patterns with characteristic diffraction peaks at the following 2θ positions: 3.92°±0.2°, 8.7°±0.2°, 15.54°±0.2° and 18.22°±0.2°.

[0067] Preferably, the crystal form IV of the compound of formula (Ib) of the present invention, when subjected to Cu-Kα radiation, has a X-ray powder diffraction pattern that further exhibits characteristic diffraction peaks at the following 2θ positions: 7.76°±0.2°, 10.48°±0.2°, 12.46°±0.2°, 16.79°±0.2°, 18.94°±0.2°, and 19.67°±0.2°.

[0068] In some embodiments, the compound represented by formula (Ib) of the present invention is of crystal form IV, and its X-ray powder diffraction pattern is substantially as shown in Figures 22-1 and / or 22-2.

[0069] In some embodiments, the compound represented by formula (Ib) of the present invention is crystal form IV, and its differential scanning calorimetry (DSC) curve is shown in Figure 23 or its thermogravimetric analysis curve is shown in Figure 24.

[0070] The present invention also provides an amorphous form of the compound shown in formula (Ib), whose X-ray powder diffraction pattern using Cu-Kα radiation is essentially as shown in Figure 25.

[0071] In some embodiments, the amorphous form of the compound of formula (Ib) of the present invention is shown in Figure 26 for differential scanning calorimetry (DSC) or in Figure 27 for thermogravimetric analysis.

[0072] The present invention also provides a method for preparing a pharmaceutically acceptable salt of the compound shown in formula (I), wherein the method includes a step of forming a salt with the compound shown in formula (I) and an acid.

[0073] In some embodiments of the method for preparing the compound represented by formula (I) of the present invention, the solvent used is selected from C 1-6 Halogenated alkane solvents, C 2-6 Ester solvents, C 2-6 Ether solvents, C 1-6 One or more of an alcohol solvent or water, preferably one or more of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropanol, diethyl ether, tetrahydrofuran, and water, more preferably one or more of dichloromethane, methanol, ethanol, and water.

[0074] In some embodiments of the method for preparing the compound represented by formula (I) of the present invention, the method includes the step of forming a salt with an acid and the compound represented by formula (Ia); wherein the acid is selected from maleic acid, fumaric acid, hydrohalic acid (preferably hydrobromic acid and hydrochloric acid), sulfuric acid, phosphoric acid, L-tartaric acid, citric acid, L-malic acid, hippuric acid, D-glucuronic acid, glycolic acid, mucoic acid, succinic acid, lactic acid, orotic acid, pamoic acid, glycine, alanine, arginine, cinnamic acid, benzoic acid, benzenesulfonic acid, p-toluenesulfonic acid, acetic acid, propionic acid, valeric acid, triphenylacetic acid, L-proline, ferulic acid, 2-hydroxyethanesulfonic acid, mandelic acid, nitric acid, methanesulfonic acid, malonic acid, gentian acid, salicylic acid, oxalic acid, or glutaric acid.

[0075] In some embodiments of the method for preparing the maleate salt of the compound represented by formula (I) of the present invention, the method includes: forming a salt with maleic acid and the compound represented by formula (Ia) to prepare the compound represented by formula (II).

[0076] The present invention also provides a method for preparing the crystal form of a compound represented by formula (Ia), (Ib) or (II), wherein the method comprises the following steps: preparing the compound of formula (II), (Ia), (Ib) with any crystal form or the amorphous compound of formula (II), (Ia), (Ib) by recrystallization or pulping; wherein the solvent for recrystallization or pulping is selected from C 2-6 Ester solvents, C 2-6 Ether solvents, C 1-6 Alcohol solvents, C 1-6 The solvent used for recrystallization or pulping is preferably one or a mixture of two or more of the following solvents: acetic acid solvent, alkane solvent, and water.

[0077] In some embodiments of the method for preparing the crystal form of the compound represented by formula (Ia), (Ib) or (II) of the present invention, the recrystallization or pulping temperature is 4 to 100°C, preferably room temperature to 90°C, and more preferably 40 to 90°C.

[0078] In some embodiments of the method for preparing crystal form I of the compound represented by formula (II) of the present invention, the method includes the following steps: mixing the compound represented by formula (II) with a suitable solvent to form a suspension, heating and stirring to form a slurry, allowing it to stand and crystallize, and filtering to separate the slurry; the solvent is preferably ethanol; the stirring temperature is preferably 90°C.

[0079] In some embodiments of the method for preparing crystal form III of the compound of formula (Ia) of the present invention, the method includes the following steps: mixing the amorphous compound of formula (Ia) with a suitable solvent, heating and stirring to form a slurry, and filtering to separate the contents; the solvent is preferably an acetonitrile / water mixed solvent; the stirring temperature is preferably 40°C.

[0080] In another aspect, the present invention also provides a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of any of the compounds or crystals described in any of the preceding claims of the present invention, and pharmaceutically acceptable excipients.

[0081] In another aspect, the present invention also provides the use of pharmaceutically acceptable salts of compounds of formula (I) or crystals and pharmaceutical compositions of compounds of formulas (Ia), (Ib), and (II) in the preparation of medicaments for the treatment and / or prevention of tumors.

[0082] In another aspect, the present invention also provides a method for treating and / or preventing tumors, the method comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt of a compound of formula (I) or a crystal or pharmaceutical composition of a compound of formulas (Ia), (Ib), or (II).

[0083] It can be understood that expressions such as "preferably, ..., its X-ray powder diffraction pattern further has characteristic diffraction peaks at the following 2θ positions", or "more preferably, ..., its X-ray powder diffraction pattern further has characteristic diffraction peaks at the following 2θ positions", etc., in this invention refer to the presence of characteristic diffraction peaks at the aforementioned 2θ positions, and further, characteristic diffraction peaks at the aforementioned "the following 2θ positions".

[0084] The X-ray powder diffraction or DSC pattern and TGA pattern disclosed in this invention, which are substantially the same, also fall within the scope of this invention.

[0085] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0086] "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.

[0087] IC 50 "Half-inhibition concentration" refers to the concentration at which half of the maximum inhibitory effect is achieved.

[0088] The "ether solvents" mentioned in this invention refer to chain or cyclic compounds containing an ether bond -O- and having 1 to 10 carbon atoms. Specific examples include, but are not limited to, tetrahydrofuran, diethyl ether, propylene glycol methyl ether, methyl tert-butyl ether, isopropyl ether, or 1,4-dioxane.

[0089] The "alcohol solvent" mentioned in this invention refers to one or more "hydroxyl groups" replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the hydroxyl group and the C 1-6 "Alkyl" as defined above, specific examples include but are not limited to: methanol, ethanol, isopropanol, n-propanol, isoamyl alcohol, or trifluoroethanol.

[0090] The "ester solvent" mentioned in this invention refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include, but are not limited to, ethyl acetate, isopropyl acetate, or butyl acetate.

[0091] The "ketone solvents" mentioned in this invention refer to compounds in which a carbonyl group (-C(O)-) is attached to two hydrocarbon groups. Depending on the hydrocarbon groups in the molecule, ketones can be classified into aliphatic ketones, alicyclic ketones, aromatic ketones, saturated ketones, and unsaturated ketones. Specific examples include, but are not limited to, acetone, acetophenone, and 4-methyl-2-pentanone.

[0092] The "nitrile solvent" mentioned in this invention refers to one or more "cyano" groups replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the "cyano" and "C" 1-6 "Alkyl" is as defined above, and specific examples include, but are not limited to, acetonitrile or propionitrile.

[0093] The "halogenated hydrocarbon solvent" described in this invention refers to one or more "halogen atoms" replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the halogen atom and the C 1-6 "Alkyl" is defined above, and specific examples include, but are not limited to, dichloromethane, 1,2-dichloroethane, chloroform, or carbon tetrachloride.

[0094] As used in this invention, "crystal of the present invention", "crystal form of the present invention", "polymorph of the present invention" and the like can be used interchangeably.

[0095] The "room temperature" mentioned in this invention generally refers to 4-30℃, and preferably to 20±5℃.

[0096] The crystal structure of this 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), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA). Thermogravimetric analysis (TGA) is also called thermogravimetry (TG).

[0097] The X-ray powder diffractometer (XRD) used in this invention is a Bruker D8 Advance diffractometer, and the copper target wavelength is [wavelength missing]. The instrument used included Kα radiation (40KV, 40mA), an θ-2θ goniometer, a Mo monochromator, a Lynxeye detector, Al2O3 as calibration material, Diffrac Plus XRD Commander as the acquisition software, and MDI Jade 6 as the analysis software. Method parameters included: a non-reflective sample plate with a diameter of 24.6 mm x a thickness of 1.0 mm (manufactured by MTIcorporation), a variable-temperature hot stage manufactured by Shanghai Micro-Touch Instrument Technology Development Co., Ltd., a copper sample plate for the variable-temperature hot stage, detection angles of 3-40°2θ / 3-30°2θ (hot stage XRPD), and a step size of 0.02°2θ.

[0098] The differential thermal analysis scanner (DSC) used in this invention is a TA Instruments Q200 DSC or DSC 3, under nitrogen protection, with a gas flow rate of 50 mL / min.

[0099] The thermogravimetric analyzer (TGA) used in this invention is a TA Instruments Q500 TGA or a TGA / DSC 3. + Nitrogen protection, with a gas flow rate of 40 mL / min or 50 mL / min.

[0100] The “2θ or 2θ angle” mentioned in this invention refers to the diffraction angle, where θ is the Bragg angle and the unit is ° or degree. The error range of the 2θ can be ±0.3, ±0.2 or ±0.1.

[0101] 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.

[0102] 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, DSC, and TGA. 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.

[0103] It is understood that, as is well known in the field of differential scanning calorimetry (DSC), the melting peak height of a DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystalline compounds of the present invention are characterized by DSC plots with characteristic peak positions, possessing substantially the same properties as the DSC plots provided in the accompanying drawings of the present invention, with an error tolerance of ±3°C.

[0104] The crystal form disclosed in this invention can be prepared using the following common methods for preparing crystal forms:

[0105] 1. The evaporation experiment involves evaporating a clear solution of the sample in an open container at different temperatures until the solvent is dry.

[0106] 2. The crystal slurry experiment involves stirring a supersaturated solution of the sample (containing insoluble solids) in different solvent systems at a certain temperature.

[0107] 3. The solvent resistance test involves dissolving the sample in a good solvent, adding the solvent, stirring the precipitated solid briefly, and then filtering it immediately.

[0108] 4. The cooling crystallization experiment involves dissolving a certain amount of sample into a corresponding solvent at high temperature, and then directly stirring and crystallizing at room temperature or low temperature.

[0109] 5. The polymer template experiment involves adding different types of polymer materials to a clear solution of the sample and leaving it at room temperature to evaporate until the solvent is dry.

[0110] 6. Thermal method experiments involve treating the sample under specific thermal crystallization conditions and then cooling it to room temperature.

[0111] 7. The water vapor diffusion experiment involves placing the sample in an environment with a certain humidity at room temperature. Attached Figure Description

[0112] Figure 1 shows the XRD pattern of the amorphous form of compound 1.

[0113] Figure 2 shows the DSC spectrum of the amorphous form of compound 1.

[0114] Figure 3 shows the TGA spectrum of the amorphous form of compound 1.

[0115] Figure 4 shows the XRD pattern of crystal form I of compound 1.

[0116] Figure 5 shows the DSC spectrum of crystal form I of compound 1.

[0117] Figure 6 shows the TGA spectrum of crystal form I of compound 1.

[0118] Figure 7 shows the XRD pattern of crystal form II of compound 1.

[0119] Figure 8 shows the DSC spectrum of crystal form II of compound 1.

[0120] Figure 9 shows the TGA spectrum of crystal form II of compound 1.

[0121] Figure 10 shows the XRD pattern of crystal form III of compound 1.

[0122] Figure 11 shows the DSC spectrum of crystal form III of compound 1.

[0123] Figure 12 shows the TGA spectrum of compound 1, crystal form III.

[0124] Figure 13-1 shows the XRD pattern of crystal form I of compound 2.

[0125] Figure 13-2 shows the XRD pattern of crystal form I of compound 2.

[0126] Figure 14 shows the DSC spectrum of crystal form I of compound 2.

[0127] Figure 15 shows the TGA spectrum of crystal form I of compound 2.

[0128] Figure 16-1 shows the XRD pattern of crystal form II of compound 2.

[0129] Figure 16-2 shows the XRD pattern of crystal form II of compound 2.

[0130] Figure 17 shows the DSC spectrum of crystal form II of compound 2.

[0131] Figure 18 shows the TGA spectrum of compound 2, crystal form II.

[0132] Figure 19-1 shows the XRD pattern of crystal form III of compound 2.

[0133] Figure 19-2 shows the XRD pattern of crystal form III of compound 2.

[0134] Figure 20 shows the DSC spectrum of compound 2, crystal form III.

[0135] Figure 21 shows the TGA spectrum of compound 2, crystal form III.

[0136] Figure 22-1 shows the XRD pattern of crystal form IV of compound 2.

[0137] Figure 22-2 shows the XRD pattern of crystal form IV of compound 2.

[0138] Figure 23 shows the DSC spectrum of crystal form IV of compound 2.

[0139] Figure 24 shows the TGA spectrum of crystal form IV of compound 2.

[0140] Figure 25 shows the XRD pattern of the amorphous form of compound 2.

[0141] Figure 26 shows the DSC spectrum of the amorphous form of compound 2.

[0142] Figure 27 shows the TGA spectrum of the amorphous form of compound 2.

[0143] Figure 28 shows the XRD pattern of crystal form I of compound 3.

[0144] Figure 29 shows the DSC spectrum of crystal form I of compound 3.

[0145] Figure 30 shows the TGA spectrum of crystal form I of compound 3.

[0146] Figure 31 shows the XRD pattern of the amorphous form of compound 3.

[0147] Figure 32 shows the DSC spectrum of the amorphous form of compound 3.

[0148] Figure 33 shows the TGA spectrum of the amorphous form of compound 3.

[0149] Figure 34 shows the XRD pattern of crystal form II of compound 3.

[0150] Figure 35 shows the DSC spectrum of crystal form II of compound 3.

[0151] Figure 36 shows the TGA spectrum of compound 3, crystal form II. Detailed Implementation

[0152] The following detailed embodiments illustrate the implementation process and beneficial effects of the present invention, aiming to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the scope of implementation of this case.

[0153] Example 1: Preparation of Compound 1

[0154] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1, also known as compound (Ia))

[0155] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

[0156]

[0157]

[0158] Step 1: 3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-carboxylic acid tert-butyl ester (1b)

[0159] tert-butyl 3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidine-1-carboxylate

[0160]

[0161] 3-(4-phenoxyphenyl)-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1a) (synthetic method see J.Med.Chem.2015,58,9625-9638) (11.0 g, 28.5 mmol) was dissolved in 100 mL of 1,2-dichloroethane. Tert-butyl 3-oxoazacyclobutane-1-carboxylate (9.74 g, 56.9 mmol) and glacial acetic acid (3.42 g, 57.0 mmol) were added sequentially. After the addition was complete, the reaction mixture was reacted at 65 °C for 3 h. The reaction solution was cooled to room temperature, and sodium triacetoxyborohydride (12.1 g, 57.1 mmol) was added. After the addition was complete, the reaction mixture was reacted overnight at room temperature. The pH of the reaction solution was adjusted to 9-10 by adding saturated sodium bicarbonate solution dropwise. After concentration under reduced pressure, the crude product was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 100:0-19:1) to obtain tert-butyl 3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobutane-1-carboxylic acid (1b) (7.20 g, yield: 47%).

[0162] LCMS m / z = 542.3[M+1] +

[0163] Step 2: 1-[1-(azacyclobut-3-yl)-4-piperidinyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine (1c)

[0164] 1-[1-(azetidin-3-yl)-4-piperidyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine

[0165]

[0166] 3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-carboxylic acid tert-butyl ester (1b) (7.20 g, 13.3 mmol) was dissolved in 15 mL of dichloromethane, 50 mL of 4N ethyl hydrochloride solution and 10 mL of anhydrous methanol were added, and the mixture was stirred at room temperature for 2 h. After the reaction solution was concentrated under reduced pressure, the residue was added to 20 mL of dichloromethane, and the pH was adjusted to 9-10 with saturated sodium bicarbonate solution. The layers were separated, and the aqueous layer was extracted with methanol / dichloromethane (v / v = 1:10) (100 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 1-[1-(azacyclobut-3-yl)-4-piperidinyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidine-4-amine (1c) (5.80 g, yield: 99%).

[0167] LCMS m / z = 442.2[M+1] +

[0168] Step 3: 3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-carboxylic acid tert-butyl ester (1d)

[0169] tert-butyl 3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidine-1-carboxylate

[0170]

[0171] 1-[1-(azacyclobut-3-yl)-4-piperidinyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine (1c) (5.80 g, 13.1 mmol) was dissolved in 25 mL of 1,2-dichloroethane. Tert-butyl 3-oxoazacyclobutane-1-carboxylate (4.50 g, 26.3 mmol) and glacial acetic acid (1.58 g, 26.3 mmol) were added sequentially. After the addition was complete, the reaction mixture was reacted at 65 °C for 3 h. The reaction solution was cooled to room temperature, and sodium triacetoxyborohydride (5.57 g, 26.3 mmol) was added. After the addition was complete, the reaction mixture was reacted overnight at room temperature. The pH of the reaction mixture was adjusted to 9-10 by adding saturated sodium bicarbonate solution dropwise. After concentration under reduced pressure, the crude product was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 100:0-19:1) to obtain tert-butyl 3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidinyl]azacyclobut-1-yl]azacyclobut-1-carboxylic acid (1d) (3.60 g, yield: 46%).

[0172] LCMS m / z = 597.3 [M+1] +

[0173] Step 4: 1-[1-[1-(azacyclobut-3-yl)azacyclobut-3-yl]-4-piperidinyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine (1e)

[0174] 1-[1-[1-(azetidin-3-yl)azetidin-3-yl]-4-piperidyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine

[0175]

[0176] 3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azolo[3,4-d]pyrimidin-1-yl]azolo[3,4-d]carboxylic acid tert-butyl ester (1d) (3.60 g, 6.03 mmol) was dissolved in 5 mL of dichloromethane, and 5 mL of trifluoroacetic acid was added. The mixture was stirred at room temperature for 2 h. After the reaction solution was concentrated under reduced pressure, the residue was added to 20 mL of dichloromethane, and the pH was adjusted to 9-10 with saturated sodium bicarbonate solution. The mixture was separated, and the aqueous layer was extracted again with 100 mL of dichloromethane. The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude 1-[1-[1-(azolo[3,4-d]pyrimidin-4-amine (1e) (3.0 g).

[0177] LCMS m / z = 497.3 [M+1] +

[0178] Step 5: 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (Compound 1)

[0179] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione

[0180]

[0181] The crude product 1-[1-[1-(azacyclobut-3-yl)azacyclobut-3-yl]-4-piperidinyl]-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-4-amine (1e) (3.00 g) was dissolved in 15 mL of dimethyl sulfoxide, and 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (synthetic method see WO2017197056) (2.00 g, 7.25 mmol) and diisopropylethylamine (3.90 g, 30.2 mmol) were added sequentially. After the addition was complete, the reaction was carried out at 90 °C for 2 h. The reaction solution was cooled to room temperature, and 10 mL of water was slowly added dropwise. The mixture was filtered, and the filter cake was dissolved in 50 mL of dichloromethane. It was then washed with 15 mL of saturated sodium chloride solution. The mixture was separated, and the organic layer was dried with anhydrous sodium sulfate. After concentration under reduced pressure, the crude product was purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 100:0-19:1). The pure product solution obtained by column chromatography was directly concentrated under reduced pressure to obtain 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (2.90 g, two-step yield from compound 1d: 64%).

[0182] Analysis using XRD, DSC, and TGA showed that compound 1 was an amorphous (yellow solid), as shown in Figures 1, 2, and 3.

[0183] 1 H NMR(400MHz, CDCl3)δ9.99(s,1H),8.39(s,1H),7.72–7.57(m,3H),7.45–7.32(m,2H),7. 21–7.11(m,3H),7.10–7.04(m,2H),6.78(d,1H),6.52(dd,1H),5.81(brs,2H),4.92(dd, 1H),4.87–4.71(m,1H),4.08–3.99(m,2H),3.94–3.83(m,2H),3.76–3.65(m,1H),3.64–3 .49(m,2H),3.20–3.04(m,3H),3.00–2.64(m,5H),2.52–2.34(m,2H),2.18–1.89(m,5H).

[0184] LCMS m / z = 377.3 [M / 2 + 1] +

[0185] Example 2: Preparation of Crystal Form I of Compound 1

[0186] 8 mL of ethyl acetate was added to the amorphous form (40 mg) of compound 1 prepared in Example 1 to dissolve it completely. The solution was then allowed to evaporate in an open container at 40 °C to obtain crystalline form I of compound 1 (yellow solid). Crystalline form I of compound 1 was characterized by XRD, DSC, and TGA, as shown in Figures 4, 5, and 6.

[0187] Example 3: Preparation of crystal form II of compound 1

[0188] 6 mL of ethanol was added to the amorphous form (200 mg) of compound 1 prepared in Example 1, and the mixture was allowed to crystallize at room temperature for 3 days. After centrifugation, the solid was dried under vacuum at room temperature overnight to obtain crystal form II of compound 1 (yellow solid). Crystal form II of compound 1 was characterized by XRD, DSC and TGA, as shown in Figures 7, 8 and 9.

[0189] Example 4: Preparation of Crystal Form III of Compound 1

[0190] 6 mL of acetonitrile and 6 mL of water were added to the amorphous form (400 mg) of compound 1 prepared in Example 1, and the mixture was stirred at 40 °C for 72 h. The mixture was then filtered, and the filter cake was collected and dried under vacuum at 40 °C overnight to obtain crystal form III of compound 1 (yellow solid). Crystal form III of compound 1 was characterized by XRD, DSC and TGA, as shown in Figures 10, 11 and 12.

[0191] Example 5: Preparation of Compound 2

[0192] 5-(3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-piperidin]-1'-yl)azacyclobut-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (compound 2, also known as compound (Ib))

[0193] 5-(3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidin]-1'-yl)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0194]

[0195] Step 1: 4-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]piperidin-1-carboxylic acid tert-butyl ester (2a)

[0196] tert-butyl 4-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]piperidine-1-carboxylate

[0197]

[0198] 3-(4-phenoxyphenyl)-1-(piperidin-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1a) (synthetic method see J. Med. Chem. 2015, 58, 9625-9638) (10.42 g, 26.97 mmol) was dissolved in 200 mL of 1,2-dichloroethane, and tert-butyl 4-oxopiperidin-1-carboxylate (13.43 g, 67.42 mmol) and glacial acetic acid (4.2 g, 67.42 mmol) were added sequentially. The mixture was heated to 65 °C and stirred for 2 hours. After cooling to room temperature, sodium triacetoxyborohydride (34.29 g, 161.79 mmol) was added, and the reaction was stirred at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the mixture was allowed to stand. 50 mL of 2M sodium hydroxide aqueous solution was added, and the pH was adjusted to 8-9 with saturated sodium bicarbonate aqueous solution. The mixture was allowed to stand and separate into layers. The aqueous phase was extracted with dichloromethane (200 mL x 3). The organic phases were combined and washed once with saturated brine (300 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (200-300 mesh silica gel, dichloromethane / methanol (v / v) = 100 / 1-15 / 1) to obtain tert-butyl 4-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]piperidin-1-carboxylic acid (2a) (10.72 g, yield: 70%).

[0199] Step 2: 3-(4-phenoxyphenyl)-1-[1-(4-piperidinyl)-4-piperidinyl]pyrazolo[3,4-d]pyrimidin-4-amine (2b)

[0200] 3-(4-phenoxyphenyl)-1-[1-(4-piperidyl)-4-piperidyl]pyrazolo[3,4-d]pyrimidin-4-amine

[0201]

[0202] 4-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]piperidin-1-carboxylic acid tert-butyl ester (2a) (45 g, 92.48 mmol) was added to a reaction flask, followed by 410 mL of dichloromethane. After stirring to dissolve, 80 mL of trifluoroacetic acid was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain an oily substance. 500 mL of dichloromethane was added, and 2 mol / L sodium hydroxide solution was slowly added dropwise under stirring to adjust the pH to 10. The aqueous phase was extracted with dichloromethane (400 mL x 3), and the organic phases were combined. The organic layer was washed with 15% sodium chloride aqueous solution (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 3-(4-phenoxyphenyl)-1-[1-(4-piperidinyl)-4-piperidinyl]pyrazolo[3,4-d]pyrimidine-4-amine (2b) (29.3 g, yield: 82%).

[0203] Step 3: 3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidine]-1'-yl) tert-butyl 3-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)azacyclobutane-1-carboxylic acid tert-butyl ester (2c)

[0204] tert-butyl 3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidin]-1'-yl)azetidine-1-carboxylate

[0205]

[0206] 3-(4-phenoxyphenyl)-1-[1-(4-piperidinyl)-4-piperidinyl]pyrazolo[3,4-d]pyrimidin-4-amine (2b) (29.3 g, 0.076 mol) was added to 1,2-dichloroethane (0.5 L), followed by the sequential addition of 1-Boc-3-azacyclobutanone (37.7 g, 0.189 mol), acetic acid (11.4 g, 0.189 mol), and anhydrous sodium sulfate (30 g). After the addition was complete, sodium triacetoxyborohydride (96.3 g, 0.45 mol) was slowly added, and the mixture was stirred at room temperature for 2 h. Pour into a 2L plastic beaker, add ice, and adjust the pH to 12-13 with 2M sodium hydroxide aqueous solution. Let stand to separate the layers. Extract the aqueous phase with dichloromethane (400mL x 3). Combine the organic phases, wash with saturated brine (600mL), dry with anhydrous sodium sulfate, filter, and concentrate under reduced pressure to obtain the crude product. Purify the crude product by column chromatography (200-300 mesh silica gel, dichloromethane / methanol (v / v) = 100 / 0-12 / 1) to obtain tert-butyl 3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidine]-1'-yl)azacyclobutane-1-carboxylic acid (2c) (35g, yield: 81%).

[0207] Step 4: 1-(1'–(azacyclobut-3-yl)-[1,4'-bipiperidine]-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine (2d)

[0208] 1-(1'-(azetidin-3-yl)-[1,4'-bipiperidin]-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine

[0209]

[0210] 25 g (0.04 mol) of tert-butyl 3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidine]-1'-yl)azacyclobutane-1-carboxylic acid tert-butyl ester (2c) was added to a reaction flask, followed by the addition of 125 mL of dichloromethane and then 50 mL of trifluoroacetic acid. After the addition was complete, the mixture was stirred at room temperature for 2 h. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain an oily substance. 200 mL of methyl tert-butyl ether was added under stirring, and a white solid gradually precipitated. The mixture was stirred at room temperature for 1 h to crystallize. After filtration, the solution was concentrated under reduced pressure to obtain 1-(1'–(azacyclobut-3-yl)-[1,4'-piperidin]-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine trifluoroacetate (2d) (50 g, yield: 99%).

[0211] Step 5: 5-(3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-piperidin]-1'-yl)azacyclobut-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (compound 2)

[0212] 5-(3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-bipiperidin]-1'-yl)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0213]

[0214] To 1-(1'–(azacyclobut-3-yl)-[1,4'-bipiperidine]-4-yl)-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine trifluoroacetate (2d) (48 g, 0.031 mol), 2-(2,6-dioxopiperidine-3-yl)-5-fluoroisoindoline-1,3-dione (synthetic method see WO2017197056) (10.3 g, 0.037 mol), N,N′-diisopropylethylamine (40 g, 0.31 mol) and dimethyl sulfoxide (0.2 L) were added sequentially, and the mixture was stirred at 120 °C for 3 h. Cool the reaction solution to room temperature with ice water, add water (0.2 L) while stirring, and a large amount of solid precipitates. Continue stirring for 30 min, filter, dry under vacuum, dissolve the filter cake with 0.5 L of dichloromethane, wash with concentrated ammonia (200 mL x 3), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue by silica gel column chromatography (dichloromethane / methanol (v / v) = 100 / 0-94 / 6). Collect the product. Ethyl acetate (0.28 L) was added to the above column-passed product, and the mixture was stirred and slurried for 20 h. The mixture was filtered, and the filter cake was dried under vacuum at 45 °C for 92 h to obtain 5-(3-(4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-[1,4'-piperidin]-1'-yl)azacyclobut-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (compound 2) (17 g, yield: 72%).

[0215] XRD, DSC and TGA analysis showed that compound 2 is crystal form I (yellow solid), as shown in Figures 13-1, 13-2, 14 and 15.

[0216] 1 H NMR (400MHz, CDCl3) δ10.22(brs,1H),8.39(s,1H),7.67–7.60(m,3H),7.42–7.34(m,2H),7.19–7.1 0(m,3H),7.10–7.04(m,2H),6.78(d,1H),6.51(dd,1H),5.89(brs,2H),4.96–4.88(m,1H),4.83–4.7 0(m,1H),4.14–4.04(m,2H),3.92–3.84(m,2H),3.39–3.30(m,1H),3.18–3.04(m,2H),3.00–2.91(m, 2H),2.90–2.65(m,3H),2.56–2.32(m,5H),2.16–2.01(m,3H),2.01–1.84(m,4H),1.73–1.59(m,2H).

[0217] LC-MS m / z = 781.4 [M+1] + .

[0218] Example 6: Preparation of crystal form II of compound 2

[0219] 2.8 mL of tetrahydrofuran and 1.4 mL of water were added to 210 mg of compound 2 in crystal form I. The mixture was heated and stirred at 60 °C until dissolved. The temperature was then lowered to 4 °C and stirred overnight. A solid precipitated out. The solid was filtered under reduced pressure and dried under vacuum at room temperature for about 3 h to obtain compound 2 in crystal form II (yellow solid). Compound 2 in crystal form II was characterized by XRD, DSC and TGA, as shown in Figures 16-1, 16-2, 17 and 18.

[0220] Example 7: Preparation of Crystal Form III of Compound 2

[0221] 14 mL of water and 1.4 mL of tetrahydrofuran were added to 210 mg of crystal form I of compound 2. The mixture was crystallized at 4 °C for 3 days and then filtered under reduced pressure to obtain crystal form III of compound 2 (yellow solid). Crystal form III of compound 2 was characterized by XRD, DSC and TGA, as shown in Figures 19-1, 19-2, 20 and 21.

[0222] Example 8: Preparation of crystal form IV of compound 2

[0223] 1 mL of isopropyl acetate and 1 mL of n-heptane were added to crystal form I (30 mg) of compound 2. The mixture was crystallized at room temperature for 3 days, centrifuged, and dried under vacuum at room temperature for about 5 hours to obtain crystal form IV (yellow solid) of compound 2. Crystal form IV of compound 2 was characterized by XRD, DSC, and TGA, as shown in Figures 22-1, 22-2, 23, and 24.

[0224] Example 9: Preparation of the amorphous form of compound 2

[0225] Add 5 mL of dichloromethane to 400 mg of crystalline form I of compound 2, heat to dissolve, filter, and concentrate the filtrate to dryness under reduced pressure at 40 °C to obtain the amorphous form of compound 2 (yellow solid). The amorphous form of compound 2 was characterized by XRD, DSC, and TGA, as shown in Figures 25, 26, and 27.

[0226] Example 10: Preparation of Compound 3

[0227] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)]pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione dimaleate (compound 3, also known as compound (II))

[0228] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione dimaleate

[0229]

[0230] Dichloromethane (10 mL) was added to 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (1.0 g, 1.33 mmol), and the mixture was stirred at room temperature until dissolved. A methanol solution of maleic acid (0.309 g, 2.66 mmol) in 1 mL was added dropwise. A solid gradually precipitated during the addition. After stirring at room temperature for 3 h, the mixture was filtered under reduced pressure. The filter cake was washed with 10 mL of dichloromethane. The collected filter cake was concentrated under reduced pressure at 40 °C to remove residual solvent, yielding 0.96 g of crude product. 20 mL of ethanol was added to the crude product, and the mixture was heated and stirred at 90 °C for 0.5 h. The suspension was cooled to room temperature and crystallized for 2 hours. The mixture was then filtered under reduced pressure. The filter cake was washed with 10 mL of ethanol. The collected filter cake was concentrated under reduced pressure at 40 °C to remove residual solvent, yielding 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)]pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione dimaleate (compound 3) (0.62 g, yield: 47%).

[0231] XRD, DSC and TGA analysis showed that compound 3 is crystal form I (yellow solid), as shown in Figures 28, 29 and 30.

[0232] 1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.27(s,1H),7.72–7.63(m,3H),7.48–7.41(m,2H),7.2 4–7.09(m,5H),6.86(d,1H),6.72(dd,1H),6.15(s,4H),5.06(dd,1H),5.01–4.86(m,1H),4.2 4–4.12(m,2H),4.11–3.93(m,3H),3.92–3.79(m,2H),3.77–3.59(m,3H),3.37–3.22(m,2H),2 .98–2.68(m,3H),2.65–2.51(m,2H),2.46–2.31(m,2H),2.18–2.06(m,2H),2.06–1.96(m,1H).

[0233] Example 11: Amorphous preparation of compound 3

[0234] 50 mL of trifluoroethanol and 50 mL of dichloromethane were added sequentially to compound 3 (300 mg) (crystal form I) to dissolve it. The solution was then concentrated under reduced pressure at 40 °C to dryness, yielding the amorphous form of compound 3 (yellow solid).

[0235] XRD, DSC and TGA analysis showed that compound 3 is amorphous, as shown in Figures 31, 32 and 33.

[0236] Example 12: Preparation of Crystal Form II of Compound 3

[0237] Add 6.0 mL of methanol and 4.0 mL of water to compound 3 (150 mg) (crystal form I), dissolve in a water bath at 70 °C, stir overnight at 4 °C, precipitate solid, filter under reduced pressure, and dry under vacuum at room temperature overnight to obtain crystal form II (yellow solid) of compound 3.

[0238] XRD, DSC and TGA analysis showed that compound 3 is crystal form II, as shown in Figures 34, 35 and 36.

[0239] Example 13:

[0240] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione L-malate (compound 4)

[0241] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione L-malate

[0242]

[0243]

[0244] Dichloromethane (8 mL) was added to 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.40 g, 0.531 mmol), and the mixture was stirred at room temperature until dissolved. A methanol solution (0.5 mL) of L-malic acid (0.28 g, 2.09 mmol) was added dropwise. A viscous solid gradually precipitated during the addition process. Stirring continued at room temperature. After 2 hours, the mixture was concentrated under reduced pressure at 40°C. Ethanol (10 mL) was added to the residue, and the mixture was heated to 90°C and stirred for 1 hour. Then, it was cooled to room temperature and stirred for 2 hours. The mixture was filtered, and the filter cake was dried under vacuum at 50°C for 18 hours to obtain 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione L-malate (compound 4) (yellow solid) (0.41 g, yield: 81%).

[0245] 1H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.24(s,1H),7.70–7.61(m,3H),7.48–7.40(m,2H),7.23 –7.08(m,5H),6.80(d,1H),6.67(dd,1H),5.05(dd,1H),4.77–4.64(m,1H),4.21(dd,1.5H),4. 11–4.02(m,2H),3.88–3.80(m,2H),3.76–3.66(m,1H),3.55–3.46(m,2H),3.16–3.04(m,3H),3 .00–2.80(m,3H),2.65–2.52(m,3H),2.49–2.38(m,2H),2.31–2.07(m,4H),2.06–1.89(m,3H).

[0246] Example 14:

[0247] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione citrate (compound 5)

[0248] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione citrate

[0249]

[0250] Dichloromethane (8 mL) was added to 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.40 g, 0.531 mmol), and stirred at room temperature until dissolved. A methanol solution (0.7 mL) of citrate monohydrate (0.45 g, 2.14 mmol) was added dropwise. A viscous solid gradually precipitated during the addition process. The reaction was continued at room temperature. After stirring for 2 hours, the mixture was concentrated under reduced pressure at 40°C. Ethanol (10 mL) was added to the residue, and the mixture was heated to 90°C and stirred for 1 hour. Then, it was cooled to room temperature and stirred for 2 hours. The mixture was filtered, and the filter cake was dried under vacuum at 50°C for 18 hours to obtain 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione citrate (compound 5) (yellow solid) (0.48 g, yield: 87%).

[0251] 1 H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.25(s,1H),7.70–7.63(m,3H),7.48–7.40(m, 2H),7.23–7.09(m,5H),6.82(d,1H),6.68(dd,1H),5.06(dd,1H),4.83–4.71(m,1H),4 .15–4.04(m,2H),3.91–3.74(m,3H),3.64–3.53(m,2H),3.33–3.20(m,3H),3.09–2.9 6(m,2H),2.94–2.82(m,1H),2.79–2.53(m,8H),2.40–2.20(m,4H),2.07–1.91(m,3H).

[0252] Example 15:

[0253] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione fumarate (compound 6)

[0254] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione fumarate

[0255]

[0256] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.500 g, 0.664 mmol) was dissolved in dichloromethane (5 mL), and anhydrous methanol (1.25 mL) and fumaric acid (0.616 g, 5.31 mmol) were added sequentially. The mixture was stirred at room temperature for 7 h, filtered, and the filter cake was collected. Anhydrous ethanol (15 mL) was added to the filter cake, and the mixture was heated to 80 °C and stirred for 2 h. Cool to room temperature, filter, and dry the filter cake under vacuum at 50 °C for 16 h to give 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione fumarate (compound 6) (yellow solid) (0.400 g, yield: 69%).

[0257] 1 H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.23(s,1H),7.70–7.60(m,3H),7.47–7.40(m,2H) ,7.23–7.09(m,5H),6.79(d,1H),6.66(dd,1H),6.62(s,2H),5.05(dd,1H),4.74–4.62(m ,1H),4.09–4.01(m,2H),3.86–3.78(m,2H),3.71–3.62(m,1H),3.50–3.40(m,2H),3.08– 2.97(m,3H),2.94–2.82(m,3H),2.64–2.46(m,2H),2.29–2.14(m,2H),2.12–1.86(m,5H).

[0258] Example 16:

[0259] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione L-tartrate (compound 7)

[0260] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione L-tartrate

[0261]

[0262] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.500 g, 0.664 mmol) was dissolved in dichloromethane (5 mL), and anhydrous methanol (0.75 mL) and L-tartaric acid (0.399 g, 2.66 mmol) were added sequentially. The mixture was stirred at room temperature for 4 h, filtered, and the filter cake was collected. Anhydrous ethanol (15 mL) was added to the filter cake, and the mixture was heated to 80 °C and stirred for 2 h. Cool to room temperature, filter, and dry the filter cake under vacuum at 50 °C for 16 h to give 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione L-tartrate (compound 7) (yellow solid) (0.510 g, yield: 79%).

[0263] 1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.24(s,1H),7.70–7.61(m,3H),7.48–7.40(m ,2H),7.23–7.09(m,5H),6.80(d,1H),6.67(dd,1H),5.06(dd,1H),4.78–4.65(m,1H ),4.28(s,3H),4.11–4.02(m,2H),3.88–3.79(m,2H),3.76–3.66(m,1H),3.56–3.40 (m,2H),3.18–3.04(m,3H),2.98–2.80(m,3H),2.65–2.46(m,2H),2.35–1.87(m,7H).

[0264] Example 17:

[0265] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione salicylate (compound 8)

[0266] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione salicylate

[0267]

[0268] To 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.400 g, 0.531 mmol), 10 mL of a mixed solvent of ethanol / water (v / v) = 4:1 was added, and the mixture was heated to 80 °C to dissolve it completely. Salicylic acid (0.293 g, 2.12 mmol) was added, and the mixture was stirred at 80 °C until the solution became clear. The temperature was lowered to 60 °C and stirred for 1 h, then cooled to room temperature and stirred for 2 h. The crystals were collected by vacuum filtration, and the filter cake was collected. 10 mL of a mixed solvent of ethanol / water (v / v) = 4 / 1 was added to the filter cake. The mixture was heated to 80 °C and stirred until the solid dissolved. The mixture was then cooled to room temperature and stirred for 2 h to crystallize. The crystals were filtered, and the filter cake was collected. The filter cake was dried under vacuum at 50 °C for 16 h to obtain 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione salicylate (compound 8) (yellow solid) (0.140 g, yield: 30%).

[0269] 1 H NMR(400MHz,DMSO-d6)δ11.05(s,1H),8.24(s,1H),7.78–7.59(m,4H),7.48–7.40(m,2H),7 .39–7.31(m,1H),7.24–7.08(m,5H),6.85–6.74(m,3H),6.65(dd,1H),5.06(dd,1H),4.87–4 .73(m,1H),4.15–4.04(m,2H),3.95–3.77(m,3H),3.70–3.58(m,2H),3.41–3.28(m,3H),3.1 4–3.01(m,2H),2.95–2.81(m,1H),2.65–2.46(m,2H),2.45–2.22(m,4H),2.07–1.94(m,3H).

[0270] Example 18:

[0271] Oxalate of 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 9)

[0272] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidyl]azetidin-1-yl]azetidin-1-yl]-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione oxalate

[0273] 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 1) (0.500 g, 0.664 mmol) was dissolved in dichloromethane (10 mL), and a methanol solution (2 mL) of oxalate dihydrate (0.335 g, 2.66 mmol) was slowly added dropwise. After stirring at room temperature for 4 h, the mixture was filtered, and the filter cake was collected. Anhydrous ethanol (12 mL) was added to the filter cake, and the mixture was heated to 80 °C and stirred for 2 h. Cool to room temperature, filter, collect the filter cake, and dry the filter cake under vacuum at 50 °C for 16 h to obtain oxalate of 5-[3-[3-[4-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]azacyclobut-1-yl]azacyclobut-1-yl]-2-(2,6-dioxo-3-piperidinyl)isodihydroindole-1,3-dione (compound 9) (yellow solid) (0.480 g).

[0274] 1 H NMR(400MHz,DMSO-d6)δ11.06(s,1H),8.26(s,1H),7.71–7.62(m,3H),7.49–7.39(m, 2H),7.24–7.09(m,5H),6.84(d,1H),6.70(dd,1H),5.06(dd,1H),4.95–4.83(m,1H),4 .21–4.09(m,2H),4.06–3.91(m,3H),3.88–3.74(m,2H),3.70–3.55(m,3H),3.30–3.1 6(m,2H),2.96–2.81(m,1H),2.74–2.46(m,4H),2.44–2.28(m,2H),2.15–1.94(m,3H).

[0275] Test case

[0276] 1. XRD analysis of compounds 1, 2, and 3

[0277] The compounds of the present invention were subjected to X-ray powder diffraction tests according to the following method. The test parameters for amorphous, crystal form I, crystal form II and crystal form III of compound 1 are shown in Table 1-1. The test parameters for crystal forms I, II, III, IV and amorphous of compound 2 are shown in Table 1-1. The test parameters for crystal forms I, II and amorphous of compound 3 are shown in Table 1-2. The test results are shown in Figures 1, 4, 7, 10, 13-1, 13-2, 16-1, 16-2, 19-1, 19-2, 22-1, 22-2, 25, 28, 31 and 34.

[0278] Table 1-1 XRD Test Parameters

[0279]

[0280]

[0281] Table 1-2 XRD Test Parameters

[0282]

[0283] 2. DSC testing of compounds 1, 2, and 3

[0284] DSC spectra were acquired using a TA Instruments Q200 DSC and DSC 3 differential thermal analyzer. The test parameters for compounds 1 and 2 are shown in Table 2-1, and the test parameters for compound 3 are shown in Table 2-2. The test results are shown in Figures 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, and 35.

[0285] Table 2-1 DSC Test Parameters

[0286]

[0287] Table 2-2 DSC Test Parameters

[0288]

[0289] 3. TGA testing of compounds 1, 2, and 3

[0290] TGA spectra on TA Instruments Q500 TGA and TGA / DSC 3 + The test parameters for compounds 1 and 2 were collected using a thermogravimetric analyzer and are shown in Table 3-1. The test parameters for compound 3 are shown in Table 3-2. The test results are shown in Figures 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33 and 36.

[0291] Table 3-1 TGA Test Parameters

[0292]

[0293] Table 3-2 TGA Test Parameters

[0294]

[0295] 4. Specific peak characterization results of XRD tests for compounds 1, 2, and 3

[0296] The X-ray powder diffraction (XRD) pattern of crystal form I of compound 1 is shown in Figure 4. Specific peak values ​​are shown in Table 4.

[0297] Table 4

[0298]

[0299]

[0300] The X-ray powder diffraction (XRD) pattern of crystal form II of compound 1 is shown in Figure 7. Specific peak values ​​are shown in Table 5.

[0301] Table 5

[0302]

[0303]

[0304] The X-ray powder diffraction (XRD) pattern of crystal form III of compound 1 is shown in Figure 10. Specific peak values ​​are shown in Table 6.

[0305] Table 6

[0306]

[0307]

[0308] The X-ray powder diffraction (XRD) patterns of crystal form I of compound 2 are shown in Figures 13-1 and 13-2. Specific peak values ​​are shown in Table 7.

[0309] Table 7

[0310]

[0311]

[0312] The X-ray powder diffraction (XRD) patterns of crystal form II of compound 2 are shown in Figures 16-1 and 16-2. Specific peak values ​​are shown in Table 8.

[0313] Table 8

[0314] 2-Theta d Height I％ Area I％ 5.116 17.2585 514 41.8 6102 41.8 6.682 13.218 1230 100 14607 100 9.986 8.8505 169 13.7 2115 14.5 11.036 8.0108 45 3.7 636 4.4 13.441 6.5823 232 18.9 3725 25.5 13.863 6.3829 206 16.7 2553 17.5 15.341 5.7711 201 16.3 3304 22.6 15.762 5.6176 148 12 2952 20.2 16.503 5.3673 424 34.5 5827 39.9 18.976 4.6729 63 5.1 650 4.4 20.182 4.3963 575 46.7 10939 74.9 20.988 4.2293 97 7.9 980 6.7 21.255 4.1766 62 5 612 4.2 22.399 3.966 204 16.6 2155 14.8 23.121 3.8436 242 19.7 5358 36.7 24.14 3.6837 82 6.7 1954 13.4 24.54 3.6245 41 3.3 538 3.7 26.281 3.3883 110 8.9 3092 21.2 26.595 3.3489 56 4.6 2211 15.1 27.321 3.2616 47 3.8 814 5.6 27.574 3.2322 40 3.3 815 5.6 28.283 3.1528 56 4.6 864 5.9 28.538 3.1252 42 3.4 879 6 30.409 2.937 35 2.8 381 2.6 31.342 2.8517 40 3.3 512 3.5

[0315] The X-ray powder diffraction (XRD) patterns of crystal form III of compound 2 are shown in Figures 19-1 and 19-2. Specific peak values ​​are shown in Table 9.

[0316] Table 9

[0317]

[0318]

[0319] The X-ray powder diffraction (XRD) patterns of crystal form IV of compound 2 are shown in Figures 22-1 and 22-2. Specific peak values ​​are shown in Table 10.

[0320] Table 10

[0321] 2-Theta d Height I％ Area I％ 3.918 22.5314 331 60.2 3600 38.7 7.762 11.3801 146 26.5 1419 15.3 8.7 10.1553 550 100 9302 100 10.136 8.7195 66 12 1716 18.4 10.481 8.4337 145 26.4 3370 36.2 12.048 7.3398 54 9.8 1454 15.6 12.46 7.0979 190 34.5 2852 30.7 13.914 6.3592 56 10.2 1100 11.8 15.542 5.6967 224 40.7 3615 38.9 16.785 5.2777 139 25.3 1469 15.8 17.508 5.0613 87 15.8 1830 19.7 18.221 4.8649 205 37.3 4139 44.5 18.943 4.681 143 26 1862 20 19.665 4.5107 183 33.3 3792 40.8 23.639 3.7605 72 13.1 1363 14.7

[0322] The X-ray powder diffraction (XRD) pattern of crystal form I of compound 3 is shown in Figure 28. Specific peak values ​​are shown in Table 11.

[0323] Table 11

[0324]

[0325]

[0326] The X-ray powder diffraction (XRD) pattern of crystal form II of compound 3 is shown in Figure 34. Specific peak values ​​are shown in Table 12.

[0327] Table 12

[0328]

[0329]

[0330] 5. Chemical stability data of compound 1 and its pharmaceutically acceptable salts

[0331] Samples were tested under high temperature (40℃) and high humidity (RH 92.5%) conditions, and the purity (expressed as a percentage) was determined by HPLC. The experimental results are shown in Table 16.

[0332] The preparation methods of the test solution and the conditions for HPLC purity detection are shown in Tables 13, 14, and 15.

[0333] Table 13 Preparation method of test sample solution 1

[0334]

[0335] Table 14. Preparation method of test sample solution 2

[0336]

[0337] Table 15 HPLC Purity Detection Conditions

[0338]

[0339]

[0340] Table 16 Chemical stability of different salts and crystal forms of compound 1 under different conditions (content determined by HPLC)

[0341]

[0342] Conclusion: The amorphous and crystalline forms III of compound 1, and the pharmaceutically acceptable salts of compound 1 (such as crystalline form I of compound 3, compound 5, and compound 9) have good chemical stability.

[0343] 6. Crystal stability data of compounds 1 and 3

[0344] 6.1. Crystal form stability of compounds 1 and 3. See Table 17.

[0345] Table 17 Crystal form stability of compounds 1 and 3

[0346]

[0347]

[0348] Conclusion: The amorphous, crystalline forms I, II, and III of compound 1 and the amorphous, crystalline form I, and crystalline form II of compound 3 have good stability.

[0349] 6.2. Room temperature competition experiments of crystal forms I, II, and III of compound 1 were conducted to investigate the crystal form stability at room temperature in isopropyl acetate and water / acetonitrile (v / v = 1:1) solvents; room temperature competition experiments of crystal forms I and II of compound 3 were conducted to investigate the crystal form stability at room temperature in ethanol / water (v / v = 1:1) and acetone / water (v / v = 1:1) solvents. The details are shown in Table 18.

[0350] Table 18 Crystal competition experiments for compounds 1 and 3

[0351]

[0352] As can be seen from the crystal competition slurry experiment of compound 1 above, crystal form III is the most stable crystal form of compound 1 at room temperature; as can be seen from the crystal competition slurry experiment of compound 3 above, crystal form I is the most stable crystal form of compound 3 at room temperature.

[0353] Solubility data in water at 7.25℃

[0354] Table 19 Solubility of different medicinal salts of Compound 1 in water at 25°C

[0355]

[0356] Conclusion: Compound 1 and its pharmaceutically acceptable salts have a certain solubility in water at 25°C. The pharmaceutically acceptable salts of Compound 1 (such as crystal form I of Compound 3, Compound 4, Compound 5, Compound 7, and Compound 9) have significantly higher solubility than Compound 1, by more than 15 times.

[0357] 8. Detection of BTK degradation in Mino cells

[0358] Mino mantle cell lymphoma cell line, purchased from ATCC, was cultured under the following conditions: RPMI-1640 + 15% FBS + 1% penicillin-drug antibody at 37°C in a 5% CO2 incubator. Cells were seeded into 6-well plates at 5 × 10⁶ cells / well. 5 Cells / well. After plating, different concentrations of compounds were added, and the cells were incubated at 37°C and 5% CO2 for 48 hours. After the culture, the cells were collected, and RIPA lysis buffer (Beyotime, Cat. P0013B) was added and lysed on ice for 15 minutes. After centrifugation at 12000 rpm and 4°C for 10 minutes, the supernatant protein sample was collected. After protein quantification using the BCA kit (Beyotime, Cat. P0009), the protein was diluted to 0.25 mg / mL. The expression of BTK (CST, Cat. 8547S) and the internal control β-actin (CST, Cat. 3700S) was detected using the fully automated Western blot quantitative analyzer (Proteinsimple) and the kit (Protein Simple, Cat. SM-W004). The expression level of BTK relative to the internal control was calculated using Compass software, and DC was calculated using Origen 9.2 software according to Equation (1). 50 Values. Among them, BTK administration represents the BTK expression level in different dosage groups, and BTK solvent represents the BTK expression level in the solvent control group.

[0359] BTK% = BTK dosage / BTK solvent × 100% Equation (1)

[0360] Table 20 BTK degradation in DCs in Mino cells 50 value

[0361] Serial Number Compound numbering <![CDATA[DC 50 (nM)]]> 1 Compound 2 22.9 2 Compound 1 10.9

[0362] Conclusion: Compounds 1 and 2 significantly degrade BTK in Mino cells.

[0363] 9. Detection of BTK protein degradation in mouse spleen

[0364] Female ICR mice, 6-8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the experiment began after 3 days of acclimatization. After being administered different doses of the compound by gavage for 3 consecutive days, the spleen of the mice was harvested, spleen cells were collected, and RIPA lysis buffer (Beyotime, Cat. P0013B) was added and lysed on ice for 15 minutes. After centrifugation at 12000 rpm and 4℃ for 10 minutes, the supernatant protein sample was collected. After protein quantification using the BCA kit (Beyotime, Cat. P0009), the protein was diluted to 0.25 mg / mL, and the expression of BTK (CST, Cat. 8547S) and the internal control β-actin (CST, Cat. 3700S) was detected using a fully automated Western blot quantitative analyzer (Proteinsimple). The expression level of BTK relative to the internal control was calculated using Compass software, and DD was calculated using Origen 9.2 software according to equation (2). 50 Value. Among them, BTK 给药 BTK expression levels in different dosage groups, BTK 溶媒 The expression level of BTK is shown in the solvent control group.

[0365] BTK% = BTK 给药 / BTK 溶媒 ×100% Equation (2)

[0366] Table 21. DD of BTK protein degradation in mouse spleen. 50 value

[0367] Serial Number Compound numbering <![CDATA[DD 50 (mg / kg)]]> 1 Compound 2 3.8 2 Compound 1 3.8

[0368] Conclusion: Compounds 1 and 2 significantly degrade BTK protein in mouse spleen.

[0369] 10. In vitro kinase detection

[0370] Kinases BTK wt (Carna, Cat. No. 08-180) and BTK C481S (Carna, Cat. No. 08-547) were prepared into a 2.5× kinase solution, and substrates FAM-P2 (GL Biochem, Cat. No. 112394) and ATP (Sigma, Cat. No. A7699-1G) were prepared into a 2.5× substrate solution. 5 μL of each compound at different concentrations was added to a 384-well plate, followed by 10 μL of the 2.5× kinase solution, and incubated at room temperature for 10 minutes. Then, 10 μL of the 2.5× substrate solution was added, and the plate was incubated at 28°C for an appropriate time. Finally, 30 μL of stop solution was added to terminate the reaction. The reaction was detected using a Caliper EZ reader2 instrument. The IC50 was calculated using XLFit exceladd-in version 5.4.0.8 software. 50 Value. The inhibition rate is calculated using equation (3), where max is the DMSO control reading, min is the negative control reading, and conversion is the compound reading.

[0371] Inhibition rate % = (max - conversion) / (max - min) * 100%. Equation (3)

[0372] The results are shown in Table 22:

[0373] Table 22 IC50 of BTK wt / C481S kinase inhibition 50 value

[0374] Serial Number Compound numbering <![CDATA[BTK C481S IC 50 (nM)]]> <![CDATA[BTK wt IC 50 (nM)]]> 1 Compound 1 8 6.3

[0375] Conclusion: Compound 1 has a significant inhibitory effect on BTK wt / C481S kinase.

[0376] 11. Canine pharmacokinetics test

[0377] Experimental Objective: This study aimed to evaluate the pharmacokinetic characteristics and bioavailability of the test substance in dogs by administering a single dose of the test substance intravenously and by gavage to Beagle dogs, determining the concentration of the test substance in canine plasma, and administering it intravenously and by gavage.

[0378] Experimental animals: Male Beagle dogs, approximately 8–11 kg, 0.5–1 week old, 6 dogs per compound. Purchased from Beijing Mars Biotechnology Co., Ltd.

[0379] Experimental method: See Table 23. On the day of the experiment, 6 Beagle dogs were randomly divided into groups according to their weight. They were fasted for 14-18 hours before administration but allowed free access to water. They were fed 4 hours after administration.

[0380] Table 23

[0381]

[0382] *Dosage is calculated based on free base.

[0383] Sampling: 1.0 ml of blood was collected via the jugular vein before and after drug administration and placed in an EDTAK2 centrifuge tube. The plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min.

[0384] Plasma collection time points for G1 & G2 groups: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, 10, 12, 24 h.

[0385] All samples were stored at -80℃ before analysis and testing. Samples were analyzed using HPLC-MS / MS.

[0386] Table 24 Pharmacokinetic parameters of the compounds in canine plasma

[0387]

[0388] *Note: The compound was administered via gavage (ig).

[0389] Conclusion: Compound 1 and its pharmaceutically acceptable salts have some oral absorption in dogs. The oral exposure of compound 3 (crystal form I), compound 5, compound 7, and compound 9 was significantly increased compared to that of compound 1, by more than two times.

Claims

1. A pharmaceutically acceptable salt of a compound of formula (I), ###0001### (I) wherein: Cy1 and Cy2 are each independently selected from piperidinyl or azetidinyl; and wherein the pharmaceutically acceptable salt is selected from the group consisting of a maleate salt, a fumarate salt, a hydrohalide salt, a sulfate salt, a phosphate salt, an L-tartrate salt, a citrate salt, an L-malate salt, a hippurate salt, a D-glucuronate salt, a glycolate salt, a mucate salt, a succinate salt, a lactate salt, an orotate salt, a pamoate salt, a glycine salt, an alanine salt, an arginine salt, a cinnamate salt, a benzoate salt, a besylate salt, a tosylate salt, an acetate salt, a propionate salt, a valerate salt, a triphenylacetate salt, an L-proline salt, a ferulate salt, a 2-hydroxyethansulfonate salt, a mandelate salt, a nitrate salt, a mesylate salt, a malonate salt, a gentisate salt, a salicylate salt, an oxalate salt, or a glutarate salt.

2. The pharmaceutically acceptable salt of claim 1, wherein the compound of formula (I) is selected from formula (la) or (lb), ###0002### (la) (lb) and wherein the pharmaceutically acceptable salt is selected from the group consisting of a maleate salt, a fumarate salt, a hydrohalide salt, a sulfate salt, a phosphate salt, an L-tartrate salt, a citrate salt, an L-malate salt, a hippurate salt, a D-glucuronate salt, a glycolate salt, a mucate salt, a succinate salt, a lactate salt, an orotate salt, a pamoate salt, a glycine salt, an alanine salt, an arginine salt, a cinnamate salt, a benzoate salt, a besylate salt, a tosylate salt, an acetate salt, a propionate salt, a valerate salt, a triphenylacetate salt, an L-proline salt, a ferulate salt, a 2-hydroxyethansulfonate salt, a mandelate salt, a nitrate salt, a mesylate salt, a malonate salt, a gentisate salt, a salicylate salt, an oxalate salt, or a glutarate salt.

3. The pharmaceutically acceptable salt of claim 2, wherein the hydrohalide salt is a hydrobromide salt and a hydrochloride salt.

4. The pharmaceutically acceptable salt of claim 2, wherein the pharmaceutically acceptable salt is selected from the group consisting of a maleate salt, a fumarate salt, an L-tartrate salt, a citrate salt, an L-malate salt, a salicylate salt, or an oxalate salt.

5. A pharmaceutically acceptable salt of a compound of formula (I) as represented by formula (II), ###0003### (II) wherein: Cy1 and Cy2 are each independently selected from piperidinyl or azetidinyl.

6. A crystalline form I of a compound of formula (II), having an X-ray powder diffraction pattern using Cu-Ka radiation with characteristic diffraction peaks at 5.96° ± 0.2°, 9.30° ± 0.2°, 11.86° ± 0.2°, 15.80° ± 0.2°, 21.75° ± 0.2°, and 23.93° ± 0.2°.

3. The pharmaceutically acceptable salt according to claim 1 or 2, wherein, 7. The crystalline form I of a compound of formula (II) of claim 6, having an X-ray powder diffraction pattern using Cu-Ka radiation further with characteristic diffraction peaks at 3.98° ± 0.2°, 7.65° ± 0.2°, 10.87° ± 0.2°, 16.88° ± 0.2°, 17.89° ± 0.2°, and 26.21° ± 0.2°.

4. The pharmaceutically acceptable salt of claim 2, wherein, 8. The crystalline form I of a compound of formula (II) of claim 7, having an X-ray powder diffraction pattern using Cu-Ka radiation further with characteristic diffraction peaks at 15.29° ± 0.2°, 17.33° ± 0.2°, 18.55° ± 0.2°, 19.21° ± 0.2°, 19.91° ± 0.2°, and 22.41° ± 0.2°.

5. The pharmaceutically acceptable salt of claim 1, wherein, ​ ​ ​ ​ 9. The crystalline Form I of the compound of formula (II) according to claim 8, having further characteristic diffraction peaks in its X-ray powder diffraction pattern measured using Cu-Ka radiation at positions 4.72°±0.2°, 9.58°±0.2°, 9.92°±0.2°, 12.85°±0.2°, 13.37°±0.2°, 13.75°±0.2°, 14.45°±0.2°, 27.37°±0.2°, 28.43°±0.2°, 30.27°±0.2°, 31.51°±0.2° and 34.21°±0.2°.

10. The crystalline Form I of the compound of formula (II) according to claim 9, having an X-ray powder diffraction pattern as shown in Figure 28 measured using Cu-Ka radiation.

11. The crystalline Form I of the compound of formula (II) according to claim 9, characterized in that, Its differential scanning calorimetry curve is shown in Figure 29 or its thermogravimetric analysis curve is shown in Figure 30.

12. A crystalline Form III of the compound of formula (la), having characteristic diffraction peaks in its X-ray powder diffraction pattern measured using Cu-Ka radiation at positions 5.02°±0.2°, 8.04°±0.2°, 16.91°±0.2°, 17.23°±0.2°, 18.19°±0.2°, 19.41°±0.2° and 20.03°±0.2°, 13. The crystalline Form III of the compound of formula (la) according to claim 12, having further characteristic diffraction peaks in its X-ray powder diffraction pattern measured using Cu-Ka radiation at positions 12.36°±0.2°, 14.60°±0.2°, 15.03°±0.2°, 15.73°±0.2°, 20.57°±0.2°, 21.31°±0.2° and 25.45°±0.2°.

14. The crystalline Form III of the compound of formula (la) according to claim 13, having further characteristic diffraction peaks in its X-ray powder diffraction pattern measured using Cu-Ka radiation at positions 5.19°±0.2°, 16.32°±0.2°, 18.75°±0.2°, 19.73°±0.2°, 21.91°±0.2°, 22.41°±0.2°, 23.48°±0.2°, 23.95°±0.2° and 26.33°±0.2°.

15. The crystalline Form III of the compound of formula (la) according to claim 14, having further characteristic diffraction peaks in its X-ray powder diffraction pattern measured using Cu-Ka radiation at positions 10.34°±0.2°, 24.85°±0.2°, 26.93°±0.2°, 27.57°±0.2°, 28.41°±0.2°, 29.59°±0.2°, 30.19°±0.2°, 31.77°±0.2°, 33.13°±0.2° and 35.75°±0.2°.

16. The crystalline form III of the compound of formula (Ia) according to claim 15, having an X-ray powder diffraction pattern as shown in Figure 10 using Cu-Ka radiation.

17. The crystalline Form III of the compound of formula (Ia) according to claim 15, characterized in that, The differential scanning calorimetry curve thereof is as shown in Figure 11 or the thermogravimetric analysis curve is as shown in Figure 12.

18. A process for the preparation of a pharmaceutically acceptable salt of claim 1, wherein, The method comprises the step of salifying the compound of formula (I) with an acid.

19. The method of making according to claim 18, wherein, The solvent used is selected from one or more of C 1-6 halogenated alkanes, C 2-6 esters, C 2-6 ethers, C 1-6 alcohols or water.

20. The method of making according to claim 19, wherein, The solvent used is selected from one or more of dichloromethane, 1,2-dichloroethane, ethyl acetate, methanol, ethanol, isopropanol, diethyl ether, tetrahydrofuran and water.

21. The method of making according to claim 20, wherein, The solvent used is selected from one or more of dichloromethane, methanol, ethanol and water.

22. The method of making according to claim 19, wherein, The method comprises: using the compound of formula (Ia) and maleic acid as raw materials to prepare the compound of formula (II) 23. A method of preparing a crystalline form of a compound of Formula (la), wherein, The crystalline form is crystalline form III of the compound of formula (Ia), and the method comprises the step of mixing the amorphous compound of formula (Ia) with a suitable solvent, heating and stirring to form a slurry, and filtering to obtain; the solvent is selected from acetonitrile / water mixed solvent 24. A method of preparing a crystalline form of a compound of Formula (II), wherein, The crystalline form is crystalline form I of the compound of formula (II), and the method comprises the step of mixing the compound of formula (II) with a suitable solvent to form a suspension, heating and stirring to form a slurry, standing for crystallization, and filtering to obtain; the solvent is selected from ethanol 25. The method of manufacturing according to claim 23 or 24, wherein, The slurry temperature is 4-100°C.

26. The method of manufacturing according to claim 23 or 24, wherein, The slurry temperature is room temperature-90°C.

27. The method of manufacturing according to claim 23 or 24, wherein, The slurry temperature is 40-90°C.

28. A pharmaceutical composition, wherein, The pharmaceutical composition contains a therapeutically effective amount of a pharmaceutically acceptable salt of the compound according to any one of claims 1-5 or a crystalline form according to any one of claims 6-17, and a pharmaceutically acceptable excipient.

29. Use of a pharmaceutically acceptable salt of the compound according to any one of claims 1-5 or a crystalline form according to claim 6 or 12 or a pharmaceutical composition according to claim 28 in the preparation of a medicament for treating and / or preventing tumors or cancers.

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