Crystal form of PARP1 inhibitor and preparation thereof

Abstract

The present disclosure provides a crystalline form of a PARP1 inhibitor 5-(4-((9-fluoro-4-oxo-4, 5-dihydropyrazolo [1, 5-a] quinoxaline-7-yl) methyl) piperazine-1-yl)-N, 6-dimethyl picolinamide, and a process for the preparation thereof. The crystalline forms are useful in the treatment or prevention of diseases or conditions responsive to PARP1 activity inhibition, such as cancer.

Description

Technical Field

[0001] This disclosure relates to the crystal form of the PARP1 inhibitor 5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxaline-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridine amide and its preparation. Background Technology

[0002] Poly(ADP-ribose) polymerase (PARP) catalyzes the use of NAD+ + Adding poly(ADP-ribose) to target protein molecules is a crucial process in DNA repair. This is essential for maintaining the integrity and stability of DNA and chromosomes, as well as ensuring the survival of mammalian cells. PARP1 catalyzes most intracellular ADP-ribose polymerization reactions, but PARP2 and other subtypes also possess this function. PARP1 knockout mice lack single-strand DNA damage repair capabilities (Krishnakumar R and Kraus WL, Mol Cell, 2010, 39(1):8-24). Meanwhile, cancer cells with DNA repair defects (such as BRCA1 (breast cancer 1) or BRCA2 (breast cancer 2) defects) are particularly sensitive to PARP inhibitors.

[0003] The catalytic domain of PARP2 is very similar to that of PARP1. PARP2 has also been found to possess similar functions to PARP1 and participate in DNA damage repair through the base excision repair (BER) mechanism (Schreiber et al., 2002 JBiol Chem277:23028-23036). Marketed PARP inhibitors (such as olaparib, niraparib, talazoparib, and rucaparib) exhibit inhibitory activity against both PARP1 and PARP2. According to clinical trial results, the therapeutic efficacy of these marketed PARP inhibitors is comparable, but their toxicity profiles vary considerably. For example, talazoparib has toxicities similar to those of chemotherapy drugs (such as hair loss). Taporabani also exhibited stronger inhibitory activity against TNKS1 / 2 (terminal anchor polymerase 1 or terminal anchor polymerase 2) than other PARP inhibitors (PARPi) in biochemical assays (Ryan et al., 2021 JBiol Chem 296:100251 / 1-100251 / 13). TNKS1 and TNKS2 share 83% sequence identity overall, and their catalytic domain sequences share 89% identity. They play roles in DNA repair, telomere maintenance, and Wnt / β-catenin signaling. Targeting PARPs other than PARP1 may be the cause of off-target toxicities (such as hair loss and diarrhea) caused by PARP inhibitors. In addition, inhibition of PARP2 activity has been found to cause hematologic toxicity (Farrés et al., 2013 Blood 122:44-54; Farrés et al., 2015 Cell Death and Differentiation 22:1144-1157). The toxicity of these PARP inhibitors limits their clinical application and their combination with other targeted therapies.

[0004] Therefore, the development of highly selective PARP1 inhibitors can reduce both mechanism-related and non-mechanism-related toxicities. Currently, several selective PARP1 inhibitors are under clinical development.

[0005] WO2023025307 discloses a compound of formula I (also referred to herein as "compound of formula (I)") 5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxaline-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridine amide as a selective inhibitor of PARP1, the contents of which are incorporated herein by reference.

[0006]

[0007] Certain crystal forms of compounds of formula (I) may possess beneficial properties, such as solubility, stability, bioavailability, impurity distribution, filtration properties, drying properties, and non-hygroscopicity, and can be more easily handled, micronized, and tableted. Furthermore, alternative or improved synthetic preparation methods are still needed, especially for large-scale, environmentally friendly production. Summary of the Invention

[0008] This disclosure provides crystal forms of 5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxaline-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridineamide (Formula I), said crystal forms including crystal form A and crystal form B.

[0009] In a first aspect, this disclosure provides a crystal form of a compound of formula (I),

[0010]

[0011] In some embodiments, the crystal form is characterized by X-ray powder diffraction (XRPD) spectra showing peaks at at least four of the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, and 21.8 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ.

[0012] In a second aspect, this disclosure provides crystal form A of the compound of formula (I),

[0013]

[0014] In some embodiments, the XRPD spectrum of crystal form A (also referred to as the crystal form of the second aspect or "second aspect") includes diffraction peaks at at least four (e.g., 4, 5, 6, 7, 8, 9, 10, or all 11) of the following positions: 5.4±0.2°2θ, 9.9±0.2°2θ, 10.8±0.2°2θ, 11.4±0.2°2θ, 13.2±0.2°2θ, 16.2±0.2°2θ, 17.7±0.2°2θ, 20.2±0.2°2θ, 20.5±0.2°2θ, 22.6±0.2°2θ, and 24.3±0.2°2θ. In some embodiments of crystal form A, the XRPD spectrum includes diffraction peaks at the following positions: 5.4 ± 0.2°2θ, 10.8 ± 0.2°2θ, 16.2 ± 0.2°2θ, and 20.2 ± 0.2°2θ. In some embodiments, the XRPD spectrum further includes at least one diffraction peak at the following positions: 13.2 ± 0.2°2θ, 20.5 ± 0.2°2θ, and 22.6 ± 0.2°2θ. In some embodiments, the XRPD spectrum further includes at least one diffraction peak at the following positions: 9.9 ± 0.2°2θ, 11.4 ± 0.2°2θ, and 17.7 ± 0.2°2θ. In some embodiments, the XRPD spectrum includes diffraction peaks at the following locations: 5.4±0.2°2θ, 9.9±0.2°2θ, 10.8±0.2°2θ, 11.4±0.2°2θ, 13.2±0.2°2θ, 16.2±0.2°2θ, 17.7±0.2°2θ, 20.2±0.2°2θ, 20.5±0.2°2θ, 22.6±0.2°2θ, and 24.3±0.2°2θ.

[0015] In some embodiments of crystal form A, the crystal form has the same characteristics as... Figure 1 The XRPD spectrum shown is essentially the same.

[0016] In some embodiments of crystal form A, the crystal form has the same characteristics as... Figure 2 The thermogravimetric analysis (TGA) curves shown are essentially the same.

[0017] In some embodiments of crystal form A, the crystal form has the same characteristics as... Figure 3 The differential scanning calorimetry (DSC) curves shown are essentially the same.

[0018] In some embodiments of crystal form A, the crystal form has at least one (e.g., one, two, or all) of the following:

[0019] (a) XRPD spectrum and Figure 1 The two are essentially the same as those shown;

[0020] (b) TGA curve and Figure 2 The two are essentially the same as those shown; and

[0021] (c) DSC curve and Figure 3 The two are essentially the same as those shown.

[0022] In some embodiments of crystal form A, the crystal form has a DSC curve showing a peak at 322.4 °C ± 5 °C.

[0023] In a third aspect, this disclosure provides crystal form B of the compound of formula (I).

[0024]

[0025] In some embodiments, crystal form B (also referred to as the third aspect crystal form or "third aspect") is characterized by an XRPD spectrum showing peaks at at least four (e.g., 4, 5, 6, 7, or all eight) of the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 13.9 ± 0.2°2θ, 21.3 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by an XRPD spectrum showing peaks at the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, and 23.5 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 12.7 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, 24.8 ± 0.2°2θ, and 26.8 ± 0.2°2θ. In some embodiments, the XRPD spectrum shows peaks at at least four (e.g., 4, 5, 6, 7, or all eight) of the following positions: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 13.9±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, and 24.8±0.2°2θ, and also shows peaks at at least one of the following positions: 12.7±0.2°2θ, 13.4±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.9±0.2°2θ, 20.1±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, and 29.5±0.2°2θ.In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.4±0.2°2θ, 13.9±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ.

[0026] In some embodiments of the third aspect, crystal form B is a hydrate, for example, crystal form B of the third aspect is a dihydrate.

[0027] In some implementations, crystal form B has the same properties as... Figure 9A The XRPD spectrum shown is essentially the same.

[0028] In some implementations, crystal form B has the same properties as... Figure 9B The XRPD spectrum shown is essentially the same.

[0029] In some implementations, crystal form B has the same properties as... Figure 10A The TGA curve shown is essentially the same.

[0030] In some implementations, crystal form B has the same properties as... Figure 10A The DSC curves shown are essentially the same.

[0031] In some implementations, crystal form B has the same properties as... Figure 10B The DSC curves shown are essentially the same.

[0032] In some implementations, crystal form B has the same properties as... Figure 10C The TGA curve shown is essentially the same.

[0033] In some embodiments of crystal form B, the crystal form has at least one (e.g., one, two, or all) of the following:

[0034] (a) XRPD and Figure 9A or Figure 9B The result (after 16 hours of drying) is essentially the same;

[0035] (b) TGA curve and Figure 10A The two are essentially the same as those shown;

[0036] (c) DSC curve and Figure 10A The two are essentially the same as those shown;

[0037] (d) DSC curve and Figure 10B The result (after 16 hours of drying) is essentially the same; and

[0038] (e) TGA curve and Figure 10C The result (after 16 hours of drying) is essentially the same.

[0039] In some embodiments of the third aspect, the crystal form has a DSC curve showing DSC endothermic peaks at any one or more (e.g., one, two, or all three) of the following temperatures: 101℃±5℃, 288℃±5℃, and 309℃±5℃, and / or endothermic onset temperatures at any one or more (e.g., one, two, or all three) of the following temperatures: 57℃±5℃, 280℃±5℃, and 295℃±5℃. In some embodiments of the third aspect, after the crystal is dried under a hot lamp, for example for 16 hours, the crystal form has a DSC curve showing DSC endothermic peaks at any one or two of the following temperatures: 83℃±5℃ and 318℃±5℃, and / or endothermic onset temperatures at any one or two of the following temperatures: 21℃±5℃ and 296℃±5℃.

[0040] In another aspect, this disclosure provides a method for preparing any of the crystalline forms or amorphous forms of the compound of formula (I) disclosed herein. In some embodiments, the method comprises dissolving the compound of formula (I) in a solvent or mixture of solvents as disclosed herein, and then precipitating the compound of formula (I) under the conditions disclosed herein. In some embodiments, the starting material is a crystalline form or mixture of crystalline forms of the compound of formula (I) (e.g., those disclosed herein), or an amorphous form (e.g., as disclosed herein), or a mixture of any combination thereof.

[0041] In one aspect, this disclosure provides a method for preparing compounds of formula (I) disclosed herein. In one embodiment, the method includes the following steps:

[0042] (a) Reduction of the (SM1) compound to obtain the (VI) compound:

[0043]

[0044] (b) Preparation of compound (V) from compounds of formula (VI) and formula (SM2) in an organic solvent in the presence of a base and a coupling agent:

[0045]

[0046] (c) Reacting the compound of formula (V) in the organic solvent under alkaline conditions to obtain the compound of formula (IV);

[0047]

[0048] (d) Reacting compound (IV) with organotin in the presence of a palladium catalyst to give compound (III):

[0049]

[0050] (e) Preparation of compound (II) from compound (III) and acylation reagent:

[0051]

[0052] (f) Preparation of compound (I) by substitution reaction between compound (II) and compound (SM3) in the presence of a base and a solvent:

[0053]

[0054] In one aspect, this disclosure provides a method for preparing the crystal form of the second aspect disclosed herein, wherein the method involves recrystallizing the compound of formula (I) in a solvent to obtain the crystal form. In one embodiment, the solvent is a mixture of acetic acid and methanol.

[0055] In one aspect, this disclosure provides a method for preparing the crystal form of the third aspect disclosed herein, said method comprising the following steps:

[0056] (a) Providing the compound of formula (I) in solid form;

[0057] (b) Optionally wash the compound of formula (I);

[0058] (c) Dissolve the compound of formula (I) in an acid, optionally wherein the acid is hydrochloric acid, 1.0N hydrochloric acid, sulfuric acid, an aqueous solution of sulfuric acid, or an aqueous solution of 0.25M sulfuric acid;

[0059] (d) Optionally wash the solution with an organic solvent, wherein the organic solvent is dichloromethane, and / or extract the solution, wherein the extraction solvent is ethyl acetate;

[0060] (e) Precipitate the compound of formula (I) by combining the solution with a base, wherein the base is, optionally, sodium bicarbonate in solid or solution form, sodium carbonate in solid or solution form, ammonia or ammonia solution;

[0061] (f) Optionally, the precipitate is dried, wherein the drying may include, for example, drying under an infrared lamp at, for example, 40°C for up to 16 hours.

[0062] In one aspect, this disclosure provides a method for preparing the crystal form of the third aspect disclosed herein.

[0063] In some implementations, the method includes the following steps:

[0064] 1. Optionally, the compound of formula (I) is washed with, for example, dichloromethane, methanol and ethyl acetate (optionally in this order), the compound of formula (I) being amorphous or crystalline or a mixture thereof; during each wash, the mixture is optionally stirred or mixed to expose the solid compound to the washing solvent before removing the solvent;

[0065] 2. Collect the solid substance and add acid, such as 1.0N hydrochloric acid, until dissolved (e.g., approximately 1 mL acid / 26.5 mg of free base of compound (I)).

[0066] 3. Extraction, for example, using approximately four times the amount of ethyl acetate (v / v) of the acid for three extractions.

[0067] 4. Add saturated sodium bicarbonate (NaHCO3) solution dropwise while stirring to precipitate crystal form B;

[0068] 5. Optionally, the solid material can be separated by filtration.

[0069] In some implementations, the method includes the following steps:

[0070] 1. Dissolve the compound of formula (I) (which may be amorphous, crystalline, or a mixture thereof) in an acid (e.g., 1.0N hydrochloric acid) until dissolved (e.g., about 1 mL of acid / 45 to 50 mg of free base of compound (I)).

[0071] 2. Add sodium carbonate (Na2CO3) dropwise with stirring until a precipitate forms (approximately 1 mL Na2CO3 / 90 to 95 mg of compound (I));

[0072] 3. Optionally, the solid material can be separated by filtration.

[0073] In some embodiments, the method includes, for example, dissolving the compound of formula (I) in sulfuric acid (e.g., an aqueous solution of sulfuric acid such as 0.25 M) at ambient temperatures (e.g., 15°C to 30°C, 18°C ​​to 28°C, 20°C to 25°C), washing with a dichloromethane solution, and adding the aqueous solution to ammonia (e.g., 1.5 M) to form a slurry. The slurry is stirred at ambient temperature for 1 hour, filtered, and dried, for example, under an infrared lamp to obtain the crystal form of the compound of formula (I).

[0074] In some embodiments, the method comprises dissolving the compound of formula (I) (e.g., 1 g) in sulfuric acid (e.g., an aqueous solution of H₂SO₄) (e.g., 0.25 M, 10 mL). The resulting clear solution is combined with ammonia (e.g., 1.5 M, 5 mL) at ambient temperature. The resulting suspension is stirred, for example, for about 1 hour, filtered, washed with H₂O (e.g., 5 mL), and dried under an infrared lamp to obtain the crystal form of the third aspect disclosed herein. The material may be dried for 16 hours. The resulting material has a water content of 7.3% by weight as determined by Karl Fischer. XRPD shows a spectrum consistent with the crystal form of the third aspect disclosed herein (one hydrate molecule contains one molecule of the compound of formula (I) and approximately two water molecules). If heated for 40 hours, the material transforms into the crystal form of the second aspect disclosed herein, with a water content of 1.5% by weight as assessed by XRPD and TGA, as determined by KF.

[0075] In some embodiments, the methods disclosed herein produce crystal forms having any of the following properties: said crystal form is characterized by XRPD spectra showing peaks at at least four (e.g., 4, 5, 6, 7, or all eight) of the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 13.9 ± 0.2°2θ, 21.3 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, and 23.5 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 12.7 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, 24.8 ± 0.2°2θ, and 26.8 ± 0.2°2θ. In some embodiments, the XRPD spectrum shows peaks at at least four (e.g., 4, 5, 6, 7, or all eight) of the following positions: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 13.9±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, and 24.8±0.2°2θ, and also shows peaks at at least one of the following positions: 12.7±0.2°2θ, 13.4±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.9±0.2°2θ, 20.1±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, and 29.5±0.2°2θ.In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.4±0.2°2θ, 13.9±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ; the crystal form has the following characteristics. Figure 9A or Figure 9B The XRPD spectrum shown is substantially the same as that shown; the crystal form has the same... Figure 10A or Figure 10C The TGA curves shown are substantially the same; the crystal form has the same... Figure 10A or Figure 10B The DSC curves shown are substantially the same; the crystal form has at least one of the following: (a) and Figure 9A or Figure 9B The XRPD shown is essentially the same as that shown in (b), and Figure 10A or Figure 10C The TGA curves shown are essentially the same, and (c) is the same as... Figure 10A or Figure 10B The DSC curves shown are substantially the same; the crystal form has DSC curves showing endothermic peaks at 101℃±5℃, 288℃±5℃, and 309℃±5℃; the crystal form has DSC curves showing endothermic onset temperatures at 57℃±5℃, 280℃±5℃, and 295℃±5℃; after drying the crystal under an infrared heat lamp, for example for 16 hours (e.g., at about 40℃), the crystal form has DSC curves showing endothermic peaks at any one or two of the following temperatures: 83℃±5℃ and 318℃±5℃ and / or endothermic onset temperatures at any one or two of the following temperatures: 21℃±5℃ and 296℃±5℃. In some embodiments, the method produces a mixture of crystal forms of the compound of formula (I) or a mixture of the amorphous form of the compound of formula (I) and one or more crystal forms.

[0076] In one aspect, this disclosure provides a composition or pharmaceutical (e.g., pharmaceutically acceptable) composition comprising any one of the crystal forms of formula (I) disclosed herein, or a combination of any one of the crystal forms of formula (I) disclosed herein, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the composition, in addition to any one of the crystal forms of formula (I) disclosed herein, or a combination of any one of the crystal forms of formula (I) disclosed herein, and a pharmaceutically acceptable carrier or excipient, comprises at least one additional therapeutic agent, such as at least one known anticancer drug or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one known anticancer drug is selected from the group consisting of: abiraterone, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, doxorubicin, darolutamide, enzalutamide, epirubicin, aclarubicin, mitoxantrone, and methylhydroxy ethinylene. ellipticine), etoposide, 5-azacytidine, gemcitabine, 5-fluorouracil, capecitabine, methotrexate, 5-fluoro-2'-deoxyuridine, fludarabine, nelarabine, ara-C, pralatrexate, prednisone, pemetrexed, hydroxyurea, thioguanine, colchicine, vinblastine, vincristine, vinorelbine, paclitaxel, ixabepiloneCabazitaxel, docetaxel, panitumumab, necitumumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, obinutuzumab, ofatumumab, rituximab, alemtuzumab, ibritumomab, tositumomab, brentuximab, daramuth subcitrate Monoclonal antibodies (daratumumab), elotuzumab, T-DM1, dinutuximab, blinatumomab, ipilimumab, Avastin (bevacizumab), Herceptin (trastuzumab), MabThera (rituximab), imatinib, gefitinib, erlotinib, ostinib, afatinib, ceritinib, alectinib, crizotinib, erlotinib, lapatinib, solutinib / rafenib lafenib, regorafenib, vemurafenib, dabrafenib, aflibercept, sunitinib, nilotinib, dasatinib, bosutinib, pratinib, brutinib, cabozantinib, lenvatinib, vandetanib, trametinib, cabitinib, axitinib, temsirolimus, idelalisib, pazopanibEverolimus, tamoxifen, letrozole, fulvestrant, mitoguanhydrazone, octreotide, retinoids, arsenic, zoledronicacid, bortezomib, carfilzomib, ixazomib, vismodegib, solidegib, denosumab, thalidomide, lenalidomide, venetoclax, aldesleukin (recombinant human interleukin-2), and sipuleucel-T (prostate cancer treatment vaccine).

[0077] In one aspect, this disclosure provides a method for preparing a composition (e.g., a pharmaceutical composition or a pharmaceutically acceptable composition), the method comprising combining a crystalline form of a compound of formula (I) disclosed herein, or a combination of crystalline forms of compounds of formula (I) disclosed herein, optionally together with an amorphous form of a compound of formula (I) disclosed herein, with a pharmaceutically acceptable excipient or carrier. In some embodiments, the method comprises combining a crystalline form disclosed herein, or a combination of crystalline forms disclosed herein, with an additional therapeutic agent (e.g., at least one known anticancer drug or a pharmaceutically acceptable salt thereof) and a pharmaceutically acceptable excipient or carrier. In some embodiments, the at least one known anticancer drug is selected from the group consisting of: abiraterone, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, doxorubicin, darolutamide, enzalutamide, epirubicin, aclarubicin, mitoxantrone, and methylhydroxy ethinylene. ellipticine), etoposide, 5-azacytidine, gemcitabine, 5-fluorouracil, capecitabine, methotrexate, 5-fluoro-2'-deoxyuridine, fludarabine, nelarabine, ara-C, pralatrexate, prednisone, pemetrexed, hydroxyurea, thioguanine, colchicine, vinblastine, vincristine, vinorelbine, paclitaxel, ixabepilone, cabazitaxelDocetaxel, panitumumab, necitumumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, obinutuzumab, ofatumumab, rituximab, alemtuzumab, ibritumomab, tositumomab, brentuximab, daratumumab mumab, elotuzumab, T-DM1, dinutuximab, blinatumomab, ipilimumab, Avastin (bevacizumab), Herceptin (trastuzumab), MabThera (rituximab), imatinib, gefitinib, erlotinib, ostinib, afatinib, ceritinib, alectinib, crizotinib, erlotinib, lapatinib, solutinib lafenib, regorafenib, vemurafenib, dabrafenib, aflibercept, sunitinib, nilotinib, dasatinib, bosutinib, pratinib, brutinib, cabozantinib, lenvatinib, vandetanib, trametinib, cabitinib, axitinib, temsirolimus, idelalisib, pazopanib, everolimusTamoxifen, letrozole, fulvestrant, mitoguanhydrazone, octreotide, retinoids, arsenic, zoledronicacid, bortezomib, carfilzomib, ixazomib, vismodegib, solidegib, denosumab, thalidomide, lenalidomide, venetoclax, aldesleukin (recombinant human interleukin-2), and sipuleucel-T (prostate cancer treatment vaccine).

[0078] In one aspect, this disclosure provides a composition prepared by the methods disclosed herein for preparing compositions.

[0079] In one aspect, this disclosure provides a method of treating or preventing a condition, the method comprising applying a crystalline form of a compound of formula (I) disclosed herein, or a combination of crystalline forms of compounds of formula (I) disclosed herein, or a composition disclosed herein, to a subject in need of such treatment or prevention. In some embodiments, the condition is a condition that responds to selective inhibition of PARP1 activity relative to PARP2 activity. In some embodiments, the condition is cancer or a tumor. In some implementations, the cancers mentioned are liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer (including, for example, small cell lung cancer), Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, gastric cancer, colon cancer, malignant pancreatic islet tumors, malignant carcinoid tumors, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary neoplasms, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocythemia, adrenocortical carcinoma, skin cancer, and prostate cancer.

[0080] In some embodiments, the treatment or prevention methods disclosed herein also include the administration of an additional therapeutic agent in combination with a crystal form of the compound of formula (I), a crystal form of the compound of formula (I), or a combination of compositions (e.g., before, during, or after administration). “Combination” includes, but is not limited to, situations in which the compound of formula (I) and the additional therapeutic agent are directed to be used in a coordinated manner even if they are not administered simultaneously or in the same composition.

[0081] In one aspect, this disclosure provides the use of crystal forms of compounds of formula (I) disclosed herein, or combinations of crystal forms of compounds of formula (I) disclosed herein, or compositions disclosed herein, for the treatment of a condition. In some embodiments, the condition is a condition that responds to selective inhibition of PARP1 activity relative to PARP2 activity. In some embodiments, the condition is cancer or tumor. In some implementations, the cancers mentioned are liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer (including, for example, small cell lung cancer), Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, gastric cancer, colon cancer, malignant pancreatic islet tumors, malignant carcinoid tumors, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary neoplasms, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocythemia, adrenocortical carcinoma, skin cancer, and prostate cancer.

[0082] In one aspect, this disclosure provides the use of crystal forms of compounds of formula (I) disclosed herein, or combinations of crystal forms of compounds of formula (I) disclosed herein, or compositions disclosed herein, in the manufacture of a medicament for treating a condition. In some embodiments, the condition is a condition that responds to selective inhibition of PARP1 activity relative to PARP2 activity. In some embodiments, the condition is cancer or a tumor. In some implementations, the cancers mentioned are liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer (including, for example, small cell lung cancer), Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, gastric cancer, colon cancer, malignant pancreatic islet tumors, malignant carcinoid tumors, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary neoplasms, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocythemia, adrenocortical carcinoma, skin cancer, and prostate cancer. In some embodiments, the drug is prepared by combining a crystal form of a compound of formula (I) disclosed herein, or a combination of crystal forms of a compound of formula (I) disclosed herein, with one or more excipients. Attached Figure Description

[0083] Figure 1 XRPD of crystal form A is shown.

[0084] Figure 2 The DSC of crystal form A is shown.

[0085] Figure 3 The TGA of crystal form A is shown.

[0086] Figure 4 The XRPD overlays of crystal form A before and after high-temperature (60°C) storage are shown. Bottom diffraction pattern: acquired before storage at 60°C; Middle diffraction pattern: acquired 10 days after storage at 60°C; Top diffraction pattern: acquired 30 days after storage at 60°C.

[0087] Figure 5 The XRPD overlays of crystal form A before and after storage at high humidity (25°C / 92.5% RH) are shown. Bottom diffraction pattern: acquired before storage at 25°C / 92.5% RH; Middle diffraction pattern: acquired after 10 days of storage at 25°C / 92.5% RH; Top diffraction pattern: acquired after 30 days of storage at 25°C / 92.5% RH.

[0088] Figure 6 The XRPD superimposed images of crystal form A before and after intense light exposure (photostability measurement) are shown. Bottom diffraction pattern: acquired before exposure to intense light; Middle diffraction pattern: acquired after exposure to intense light; Top diffraction pattern: acquired after the sample wrapped in aluminum foil was exposed to intense light.

[0089] Figure 7 The XRPD of crystal form A is shown in a stacked plot after accelerated stability assessment (40 °C / 75% RH). From bottom to top, the plot shows the XRPD of the initial sample (Init.), the sample collected after 1 month of storage at 40 °C / 75% RH (1M), the sample collected after 3 months of storage at 40 °C / 75% RH (3M), and the sample collected after 6 months of storage at 40 °C / 75% RH (6M).

[0090] Figure 8 The XRPD of crystal form A is shown in a stacked plot after long-term stability assessment (25°C / 60% RH). From top to bottom, the plot shows the XRPD of the sample (1M) collected after 1 month of storage at 25°C / 60% RH, the sample (3M) collected after 3 months of storage at 25°C / 60% RH, the sample (6M) collected after 6 months of storage at 25°C / 60% RH, the sample (9M) collected after 9 months of storage at 25°C / 60% RH, and the initial sample (Init.).

[0091] Figure 9A XRPD of crystal form B in its undried state is shown.

[0092] Figure 9B The XRPD of crystal form B is shown after drying under an infrared lamp for 16 hours.

[0093] Figure 10A The DSC (bottom (green) line) and TGA (top (blue) line) of crystal form B are shown in the undried state.

[0094] Figure 10B The DSC (blue line) of crystal form B is shown after drying under an infrared lamp for 16 hours.

[0095] Figure 10C The image shows TGA of crystal form B after drying under an infrared lamp for 16 hours.

[0096] Figure 11 The asymmetric unit cell of a single-crystal structure model of amorphous type A is shown.

[0097] Figure 12 A schematic diagram of the molecular packing structure of a single crystal model of amorphous type A is shown (viewed along the a-axis of the crystal).

[0098] Figure 13 The asymmetric unit cell of the single-crystal structure model of hydrated crystal form A is shown.

[0099] Figure 14 A schematic diagram of the molecular packing structure of a single crystal model of hydrated crystal form A is shown (viewed along the a-axis of the crystal).

[0100] Figure 15 Simulated XRPD of single-crystal experiments for anhydrous form A (top: red diffraction pattern) and hydrate form A (bottom: blue diffraction pattern) is shown.

[0101] Figure 16 The XRPD comparison of the anhydrous form (red diffraction pattern) and hydrate (blue diffraction pattern) of crystal form A at 22 to 31 degrees is shown.

[0102] Figure 17 The structural differences between the fully dehydrated and partially dehydrated forms are shown.

[0103] Figure 18 The XRPD diagram of the amorphous compound of formula (I) is shown.

[0104] Figure 19 Polarized light micrograph (PLM) of the amorphous compound of formula (I) is shown.

[0105] Figure 20 XRPD of crystal form A is shown. Detailed Implementation

[0106] I. Overview

[0107] In the following description, certain specific details are set forth in order to provide a full understanding of the various embodiments of this disclosure. However, those skilled in the art will understand that this disclosure may be practiced even without these details. The following description of several embodiments is based on the understanding that this disclosure is to be regarded as instances of the claimed subject matter and is not intended to limit the appended claims to the specific embodiments shown. Headings used throughout this disclosure are provided for convenience only and should not be construed as limiting the claims in any way. Embodiments shown under any heading may be combined with embodiments shown under any other heading.

[0108] II. Definition

[0109] When an aspect or implementation is described as containing certain elements or features, it should be understood that this disclosure also discloses a corresponding aspect or implementation that is composed of those same elements or features or substantially composed of those same elements or features as an alternative aspect or implementation.

[0110] Throughout this specification, references to "an embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of this disclosure. Therefore, the phrases "in an embodiment" or "in an embodiment" appearing throughout this specification do not necessarily refer to the same embodiment. Furthermore, a particular feature, structure, or characteristic may be combined in any suitable manner in one or more embodiments.

[0111] "Pharmaceutical acceptable excipients" include, but are not limited to, adjuvants, carriers, glidants, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers approved by the U.S. Food and Drug Administration (USFDA), the China National Medical Products Administration (CNNMPA), or other relevant agencies for use in humans or livestock.

[0112] "Pharmaceutical composition" means a formulation comprising an active ingredient (e.g., a compound of formula (I) of this disclosure and its various forms (amorphous and crystalline)) and a vehicle generally accepted in the art for the delivery of a biologically active compound to a mammal (such as a human). Such vehicles include all pharmaceutically acceptable excipients for this purpose.

[0113] Unless otherwise stated, as used herein, “treatment” of a subject’s (e.g., a person, pet, domestic animal, or veterinary animal) disease, disorder, or symptom means reversing, reducing, or inhibiting the progression of the subject’s disease, disorder, or symptom, or one or more symptoms of the subject’s disease, disorder, or symptom.

[0114] As used herein, the term “prevention” of a subject’s (e.g., a person, pet, domestic animal, or veterinary animal) disease, disorder, or symptom means any treatment given to the subject that prevents the subject from developing one or more clinical symptoms of the disease, disorder, or symptom, or from developing the disease, disorder, or symptom.

[0115] The “about” value or parameter mentioned herein includes (and describes) embodiments of that value or parameter itself. For example, a description of “about X” includes a description of “X”. Furthermore, unless the context explicitly indicates otherwise, the singular forms “an” and “a” include a plural referent. Thus, for example, a reference to “the compound” includes a plurality of such compounds, and a reference to “the assay” includes a reference to one or more assays known to those skilled in the art, as well as their equivalents.

[0116] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" means, to a reasonable extent of medical judgment, a substance suitable for exposure to human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and with a reasonable benefit / risk ratio. Therefore, pharmaceutically acceptable refers to a non-biological or undesirable substance, i.e., a substance that can be administered to an individual in conjunction with the associated active compound without causing clinically unacceptable biological effects or interacting in a harmful manner with any of the other components of a pharmaceutical composition containing it.

[0117] When referring to, for example, XRD or XRPD spectra, DSC thermographs, DVS plots, or TGA, the term “substantially identical to that shown” includes spectra, thermographs, or plots that are not necessarily identical to those described herein, but which, to a person skilled in the art, fall within the limits of experimental error or deviation.

[0118] In this disclosure, the permissible error for the angular position (2θ) of the characteristic powder X-ray diffraction peak positions of a crystal form is ±0.2°. This error is used when comparing two powder X-ray diffraction patterns. If a diffraction peak in one pattern is designated as a certain angular position range of ±0.2° (2θ) of the measured peak position, and a diffraction peak in another pattern is designated as another angular position range of ±0.2° (2θ) of the measured peak position, and if these peak ranges overlap, then the two peaks are considered to have the same angular position (2θ). For example, if the diffraction peak in one pattern is determined to be at 5.20°, then for comparison, the permissible error allows the peak to be designated in the range of 5.00°–5.40°. If a control peak in another diffraction pattern is determined to be at 5.35°, then for comparison, the permissible error allows the peak to be designated in the range of 5.15°–5.55°. Because the two peak position ranges overlap, the two compared peaks are considered to have the same angular position (2θ). In some implementations, the tolerance for angular position (2θ) is ±0.1°.

[0119] III. Crystal Form

[0120] In one aspect, this disclosure provides a crystal form of a compound of formula (I),

[0121]

[0122] The crystal form is characterized by X-ray powder diffraction (XRPD) spectra showing peaks at at least four of the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, and 21.8 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, and 29.0 ± 0.2°2θ. In some embodiments, the crystal form is characterized by XRPD spectra showing peaks at the following locations: 10.8 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 20.2 ± 0.2°2θ, 21.8 ± 0.2°2θ, and 29.0 ± 0.2°2θ.

[0123] This disclosure provides a crystal form of the compound of formula (I) (5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxaloline-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridineamide). The crystal forms of this disclosure have properties suitable for medical or pharmaceutical use, including but not limited to bioavailability, stability, purity, and / or manufacturability. Specifically, this disclosure provides crystal form A of the compound of formula (I):

[0124]

[0125] In some embodiments, the crystalline form A of the compound of formula (I) of this disclosure has an X-ray powder diffraction (XRPD) spectrum including diffraction peaks (characteristic peaks) at at least four of the following positions: 5.4 ± 0.2°2θ, 9.9 ± 0.2°2θ, 10.8 ± 0.2°2θ, 11.4 ± 0.2°2θ, 13.2 ± 0.2°2θ, 16.2 ± 0.2°2θ, 17.7 ± 0.2°2θ, 20.2 ± 0.2°2θ, 20.5 ± 0.2°2θ, 22.6 ± 0.2°2θ, and 24.3 ± 0.2°2θ. In some embodiments, the XRPD spectrum of the crystalline form includes diffraction peaks at the following positions: 2θ = 5.4 ± 0.2°, 10.8 ± 0.2°, 16.2 ± 0.2°, and 20.2 ± 0.2°. In some embodiments, the XRPD spectrum of the crystal form further includes at least one diffraction peak at the following positions: 13.2±0.2°, 20.5±0.2°, and 22.6±0.2°. In some embodiments, the XRPD spectrum of the crystal form further includes at least one diffraction peak at the following positions: 9.9±0.2°, 11.4±0.2°, and 17.7±0.2°.

[0126] In some embodiments, the crystal form A of the compound of formula (I) has an XRPD spectrum including diffraction peaks at the following positions: 2θ = 5.4°±0.2°, 9.9±0.2°, 10.8±0.2°, 11.4±0.2°, 13.2°±0.2°, 16.2±0.2°, 17.7±0.2°, 20.2±0.2°, 20.5±0.2°, 22.6±0.2° and 24.3±0.2°2θ.

[0127] In some embodiments, the diffraction peak at 16.2±0.2° includes two diffraction peaks at 16.2±0.04° and 16.3±0.04°.

[0128] In some embodiments, the crystal form A of the compound of formula (I) also includes any one, any two, or all three diffraction peaks at positions of 21.0±0.2°, 26.1±0.2°, and 29.0±0.2°.

[0129] In some embodiments, the crystal form A of the compound of formula (I) has the same properties as... Figure 1 The XRPD spectra shown are essentially the same.

[0130] In some embodiments, the crystal form A of the compound of formula (I) has a diffraction peak at 2θ that is substantially the same as that shown in Table 6.

[0131] In some embodiments, the crystal form A of the compound of formula (I) has the same properties as... Figure 2The DSC thermograms shown are essentially the same.

[0132] In some embodiments, the crystal form A of the compound of formula (I) has the same properties as... Figure 3 The TGA shown is essentially the same.

[0133] In one embodiment, the crystal form A of the compound of formula (I) of this disclosure has at least one, any two, or all three of the following (a) to (c):

[0134] (a) XRPD spectrum and Figure 1 The two are essentially the same as those shown;

[0135] (b) DSC thermogram and Figure 2 The two are essentially the same as those shown; and

[0136] (c) TGA and Figure 3 The two are essentially the same as those shown.

[0137] In one aspect, this disclosure provides a crystal form of a compound of formula (I),

[0138]

[0139] The crystal form (crystal form B) is characterized by an XRPD spectrum showing peaks at at least four of the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 13.9 ± 0.2°2θ, 21.3 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by an XRPD spectrum showing peaks at the following positions: 4.6 ± 0.2°2θ, 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, and 23.5 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, and 24.8 ± 0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 9.3 ± 0.2°2θ, 10.8 ± 0.2°2θ, 12.7 ± 0.2°2θ, 21.9 ± 0.2°2θ, 23.5 ± 0.2°2θ, 24.8 ± 0.2°2θ, and 26.8 ± 0.2°2θ. In some embodiments, the XRPD spectrum shows peaks at at least four (e.g., 4, 5, 6, 7, or all eight) of the following positions: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 13.9±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, and 24.8±0.2°2θ, and also shows peaks at at least one of the following positions: 12.7±0.2°2θ, 13.4±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ. In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.9±0.2°2θ, 20.1±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, and 29.5±0.2°2θ.In some embodiments, crystal form B is characterized by XRPD spectra showing peaks at the following locations: 4.6±0.2°2θ, 9.3±0.2°2θ, 10.8±0.2°2θ, 12.7±0.2°2θ, 13.4±0.2°2θ, 13.9±0.2°2θ, 15.4±0.2°2θ, 16.1±0.2°2θ, 20.1±0.2°2θ, 21.3±0.2°2θ, 21.9±0.2°2θ, 23.5±0.2°2θ, 24.8±0.2°2θ, 26.8±0.2°2θ, 29.0±0.2°2θ, and 29.5±0.2°2θ.

[0140] In one aspect, this disclosure provides a crystal form of a compound of formula (I):

[0141]

[0142] The crystal form described therein has the same Figure 9A or Figure 9B The XRPD spectrum shown is essentially the same (after drying).

[0143] In one aspect, this disclosure provides a crystal form of a compound of formula (I):

[0144]

[0145] The crystal form described therein has the same Figure 10A or Figure 10C The TGA curve shown is essentially the same (after drying).

[0146] In one aspect, this disclosure provides a crystal form of a compound of formula (I):

[0147]

[0148] The crystal form described therein has the same Figure 10A or Figure 10B The DSC curves shown are essentially the same (after drying).

[0149] In one aspect, this disclosure provides a crystal form of a compound of formula (I):

[0150]

[0151] The crystal form said to have at least one of the following:

[0152] (a) XRPD and Figure 9A (Not dried) or Figure 9B The result (after drying with an infrared lamp for 16 hours) is essentially the same as shown;

[0153] (b) TGA curve and Figure 10A The two are essentially the same as those shown;

[0154] (c) DSC curve and Figure 10A The two are essentially the same as those shown;

[0155] (d) DCS curve and Figure 10B The result (after drying with an infrared lamp for 16 hours) is essentially the same as shown; and

[0156] (e) TGA curve and Figure 10C The result (after drying with an infrared lamp for 16 hours) is essentially the same as shown.

[0157] In one aspect, this disclosure provides a crystal form of a compound of formula (I):

[0158]

[0159] The DSC curve of the crystal form shows DSC endothermic peaks at any one or more (e.g., one, two, or all three) of the following temperatures: 101℃±5℃, 288℃±5℃, and 309℃±5℃, and / or any one or more (e.g., one, two, or all three) endothermic onset temperatures below: 57℃±5℃, 280℃±5℃, and 295℃±5℃. In some embodiments, after the crystal is dried under a hot lamp, for example, for 16 hours, the DSC curve of the crystal form shows DSC endothermic peaks at any one or two of the following temperatures: 83℃±5℃ and 318℃±5℃, and / or endothermic onset temperatures at any one or two of the following temperatures: 21℃±5℃ and 296℃.

[0160] IV. Pharmaceutical Formulations and Routes of Administration

[0161] The compounds and compositions of the present invention can be delivered directly or, together with suitable carriers or excipients, as pharmaceutical compositions or pharmaceuticals, as is well known in the art. Treatment methods of the present invention may include administering an effective amount of the compounds of the present invention to a subject in need. In a preferred embodiment, the subject is a mammalian subject, and in a most preferred embodiment, the subject is a human subject.

[0162] The effective amount of such compound, composition, or drug, as well as the most effective and convenient route of administration and the most suitable formulation, can be readily determined through routine experiments. Various formulations and drug delivery systems are available in the art. See, for example, Gennaro, AR ed. (1995) Remington's Pharmaceutical Sciences, above.

[0163] Suitable routes of administration may include, for example, oral, rectal, local, nasal, pulmonary, ocular, intestinal, and parenteral administration. Primary routes of parenteral administration include intravenous, intramuscular, and subcutaneous administration. Secondary routes of administration include intraperitoneal, intra-articular, intra-articular, intracardiac, intracisional, intradermal, intralesional, intraocular, intrapleural, intrathecal, intrauterine, and intraventricular administration. The indications for treatment and the physical, chemical, and biological properties of the drug determine the type of formulation and route of administration used, as well as whether local or systemic delivery is preferred.

[0164] The pharmaceutical dosage forms of the compounds of this invention can be provided in the form of transient release, controlled release, sustained release, or targeted drug delivery systems. Commonly used dosage forms include, for example, solutions and suspensions, (micro)emulsions, ointments, gels and patches, liposomes, tablets, lozenges, soft-shell or hard-shell capsules, suppositories, egg preparations, implants, amorphous or crystalline powders, aerosols, and lyophilized formulations. Depending on the route of administration used, special devices may be required to apply or deliver the drug, such as syringes and needles, inhalers, pumps, injection pens, applicators, or special flasks. Pharmaceutical dosage forms typically consist of a drug, excipients, and a container / closure system. One or more excipients (also called inactive ingredients) may be added to the compounds of this invention to improve or facilitate the manufacture, stability, administration, and safety of the drug, and to provide a means of achieving the desired drug release characteristics. Therefore, the type of excipient added to the drug depends on a variety of factors, such as the physical and chemical properties of the drug, the route of administration, and the manufacturing process. Pharmaceutically acceptable excipients are those available in the art and include those listed in various pharmacopoeias. See, for example, the United States Pharmacopeia (USP), the Japanese Pharmacopoeia (JP), the European Pharmacopoeia (EP), and the British Pharmacopoeia (BP); publications of the Center for Drug Evaluation and Research (CEDR) of the U.S. Food and Drug Administration (www.fda.gov), such as the Inactive Ingredient Guide (1996); Handbook of Pharmaceutical Additives, edited by Ash and Ash (2002), Synapse Information Resources, Inc., Endicott NY; etc.

[0165] The pharmaceutical dosage forms of the compounds of the present invention can be manufactured by any of the methods well known in the art (e.g., conventional mixing, sieving, dissolving, melting, granulation, tablet manufacturing, tableting, suspension, extrusion, spray drying, grinding, emulsification, (nano / micro) encapsulation, embedding, or lyophilization). As described above, the compositions of the present invention may contain one or more physiologically acceptable inactive ingredients that facilitate the processing of active molecules into pharmaceutical preparations.

[0166] Appropriate formulations depend on the desired route of administration. For intravenous injection, for example, the composition can be formulated as an aqueous solution, using physiologically compatible buffers (including, if necessary, phosphates, histidine, or citrates to adjust the pH of the formulation) and tonics (e.g., sodium chloride or glucose). For transmucosal or nasal administration, semi-solid, liquid formulations, or patches may be preferred, and they may contain penetration enhancers. Such penetration enhancers are generally known in the art. For oral administration, the compound can be formulated as a liquid or solid dosage form, as well as an instantaneous release or controlled release / sustained release formulation. Dosage forms suitable for oral ingestion by subjects include tablets, pills, lozenges, hard-shell and soft-shell capsules, liquids, gels, syrups, slurries, suspensions, and emulsions. The compound can also be formulated as rectal compositions, such as suppositories or retention enemas, for example, containing a conventional suppository base (such as cocoa butter or other glycerides).

[0167] Solid oral dosage forms can be obtained using excipients, which may include fillers, disintegrants, binders (dry and wet), dissolution retardants, lubricants, flow aids, anti-sticking agents, cation exchange resins, wetting agents, antioxidants, preservatives, colorants, and flavoring agents. These excipients can be synthetic or of natural origin. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatin, magnesium carbonate, magnesium lauryl sulfate / sodium lauryl sulfate, mannitol, polyethylene glycol, polyvinylpyrrolidone, silicates, silica, sodium benzoate, sorbitol, starch, stearic acid or its salts, sugars (i.e., glucose, sucrose, lactose, etc.), talc, tragacanth gum, vegetable oils (hydrogenated), and waxes. Ethanol and water can be used as granulation aids. In some cases, coating tablets with, for example, a taste-masking film, an acid-resistant film, or a delayed-release film is desirable. Combinations of natural and synthetic polymers with colorants, sugars, organic solvents, or water are commonly used to coat tablets, thus creating lozenges. When capsules are preferred over tablets, drug powders, suspensions, or solutions can be delivered via compatible hard-shell or soft-shell capsules.

[0168] In one embodiment, the compounds of the present invention can be applied topically, such as via skin patches, semi-solid or liquid formulations, such as gels, (micro)emulsions, ointments, solutions, (nano / micro) suspensions, or foams. The penetration of the drug into the skin and underlying tissues can be modulated, for example, by using penetration enhancers; appropriate selection and combination of lipophilic, hydrophilic, and amphiphilic excipients, including water, organic solvents, waxes, oils, synthetic and natural polymers, surfactants, emulsifiers; by pH adjustment; and by the use of complexing agents. Other techniques, such as iontophoresis, can be used to modulate the skin permeability of the compounds of the present invention. For example, transdermal or topical application is preferred where local delivery is desired and systemic exposure is minimized.

[0169] For inhalation or nasal administration, the compounds used according to the invention can be conveniently delivered from pressurized packaging or nebulizers in the form of solutions, suspensions, emulsions, or semi-solid aerosols, typically using a propellant such as methane and ethane, halocarbons derived from carbon dioxide, or any other suitable gas. For localized aerosols, hydrocarbons such as butane, isobutene, and pentane are useful. For pressurized aerosols, the appropriate dosage unit can be determined by providing a valve for dispensing the correct amount of product. Gelatin capsules and cartridges, for example, can be formulated for inhalers or blowpipes. These capsules and cartridges typically contain a powder mixture of the compound and a suitable powder base (such as lactose or starch).

[0170] Compounds and compositions formulated for parenteral administration are typically sterile and may be available in unit dosage forms, such as ampoules, syringes, injection pens, or multi-dose containers, which typically contain preservatives. Compositions may take the form of suspensions, solutions, or emulsions in oily or aqueous media and may contain formulation agents such as buffers, tonics, thickeners, surfactants, suspending and dispersing agents, antioxidants, biocompatible polymers, chelating agents, and preservatives. Depending on the injection site, the media may contain water, synthetic or vegetable oils, and / or organic cosolvents. In some cases, such as lyophilized products or concentrates, the parenteral formulation is reconstituted or diluted prior to administration. Reservoir formulations providing controlled-release or sustained-release of the compounds of the present invention may comprise injectable suspensions of nano / micron particles or nano / micron or non-micronized crystals. Polymers such as poly(lactic acid), poly(glycolic acid), or copolymers thereof, in addition to other polymers well known in the art, may also be used as controlled-release / sustained-release matrices. Other storage delivery systems can also exist in the form of implants and pumps that require incisions.

[0171] Intravenous loads suitable for the compounds of the present invention are well known in the art, including aqueous solutions containing a base (e.g., sodium hydroxide) to form ionic compounds; sucrose or sodium chloride as a tonic agent; and buffers, such as buffers containing phosphate or histidine. Cosolvents, such as polyethylene glycol, may be added. These aqueous systems effectively dissolve the compounds of the present invention and produce low toxicity upon systemic administration. The proportions of the components in the solution system can be significantly altered without compromising solubility and toxicity characteristics. Furthermore, the identities of the components can vary. For example, low-toxicity surfactants (such as polysorbates or poloxamer) may be used, as may polyethylene glycol or other cosolvents; biocompatible polymers (such as polyvinylpyrrolidone) may be added; and other sugars and polyols may be used instead of glucose.

[0172] A variety of techniques well known in the art can be used to initially estimate the effective therapeutic dose. The initial dose used in animal studies can be based on the effective concentration determined in cell culture assays. A dose range suitable for human subjects can be determined, for example, using data obtained from animal studies and cell culture assays. In some embodiments, the compounds of this disclosure are formulated for oral administration. An exemplary dose of the compounds of this disclosure in a pharmaceutical formulation for oral administration is about 0.5 to about 10 mg / kg of subject body weight. In some embodiments, the pharmaceutical formulation contains about 0.7 to about 5.0 mg / kg of subject body weight, or about 1.0 to about 2.5 mg / kg of subject body weight. Typical dosing regimens for oral administration are three times a week, twice a week, once a week, or once daily administration of the oral pharmaceutical formulation.

[0173] An effective amount or therapeutically effective amount or dose of a pharmaceutical agent (e.g., the compounds of the present invention) refers to the amount of the agent or compound that improves symptoms or prolongs survival in a subject. The toxicity and therapeutic efficacy of such molecules can be determined using standard pharmaceutical procedures in cell cultures or laboratory animals, for example, by determining the LD50 (the dose that is lethal to 50% of the population) and ED50 (the dose that is therapeutically effective to 50% of the population). The dose ratio of toxicity to therapeutic effect is the therapeutic index, which can be expressed as the LD50 / ED50 ratio. Pharmaceutical agents exhibiting a high therapeutic index are preferred.

[0174] An effective or therapeutically effective dose is the amount of a compound or pharmaceutical composition that will elicit a biological or medical response in a tissue, system, animal, or human as sought by an investigator, veterinarian, physician, or other clinician. The dose specifically falls within the cyclic concentration range, including the ED50, and has little or no toxicity. The dose may vary within this range depending on the dosage form and / or route of administration. The exact formulation, route of administration, dose, and dose interval should be selected according to methods known in the art and taking into account the specific condition of the subject.

[0175] The dosage and interval can be individually adjusted to provide a plasma level of the active component sufficient to achieve the desired effect; that is, the minimum effective concentration (MEC). The MEC varies for each compound but can be estimated, for example, through in vitro data and animal studies. The dose required to achieve the MEC depends on individual characteristics and the route of administration. In the case of local application or selective absorption, the effective local concentration of a drug may be independent of its plasma concentration.

[0176] The amount of compound or composition applied can depend on a variety of factors, including the sex, age and weight of the subject being treated, the severity of the disease, the method of administration and the judgment of the prescribing physician.

[0177] If desired, the compounds and compositions of the present invention can be contained in a packaging or dispensing device comprising one or more unit dosage forms containing an active ingredient. Such packaging or device may, for example, comprise metal or plastic foil, such as blister packs; or glass and rubber stoppers, such as vials. The packaging or dispensing device may be accompanied by instructions for use. Compositions comprising the compounds of the present invention can also be prepared, formulated with a compatible pharmaceutical carrier, placed in a suitable container, and labeled for the treatment of a specified condition.

[0178] Given the disclosure herein, those skilled in the art will readily conceive of these and other embodiments of the invention, and these and other embodiments are specifically envisioned.

[0179] V. Preparation Method

[0180] This disclosure provides a method for preparing compound (I) 5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxalin-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridineamide. The method described herein does not require any column separation or purification, has a high yield, and is particularly suitable for large-scale industrial production.

[0181] Specifically, the compound of formula (I) is prepared using a method including the following step (6):

[0182]

[0183] The compound of formula II and the compound of formula SM3 or their salts are subjected to a substitution reaction in the presence of a base and a solvent to obtain the compound of formula (I).

[0184] In the above reaction, the base comprises an inorganic base, an organic base, or a mixture of an inorganic base and an organic base. Suitable inorganic bases are selected from one or more of KI, NaI, Na₂CO₃, K₃PO₄, NaOH, and NaOAc, and suitable organic bases are selected from one or more of Et₃N (triethylamine), pyridine, DBU (1,8-diazabicycloundecane-7-ene), TMG (tetramethylguanidine), DABCO (triethylenediamine), DIEA (N,N-diisopropylethylamine), and NMM (methylmorpholine). In some embodiments, the molar ratio of inorganic base to organic base is 1:(10 to 30). In some embodiments, the base is a mixture of KI and DIEA. In some embodiments, the molar ratio of KI to DIEA is 1:(23 to 28).

[0185] In the above reaction, in some embodiments, the molar ratio of the compound of formula II to the base is 1:(3 to 7). In some embodiments, 1 mole of the compound of formula II uses about 0.1 to 0.3 moles of KI and 4.8 to 5.2 moles of DIEA. In some embodiments, 1 mole of the compound of formula II uses about 0.2 moles of KI and about 5.0 moles of DIEA.

[0186] In the above reaction, the solvent in some embodiments is selected from one or more of the following: 1,4-dioxane, water, N-methyl-2-pyrrolidone (NMP), for example, a mixture of 1,4-dioxane, water, and NMP. In some embodiments, the volume ratio of 1,4-dioxane, water, and NMP is (3.0 to 4.0):1:(3.0 to 4.0), for example (3.4 to 3.6):1:(3.2 to 3.4).

[0187] In some embodiments of the above reaction, the temperature is 95 to 105°C.

[0188] In the above reaction, in some embodiments, the molar ratio of the compound of formula II to the compound of formula SM3 or a salt thereof is 0.6 to 1.0, for example, 0.78 to 0.87. In some embodiments, the salt of the compound of formula SM3 is a hydrochloride salt.

[0189] The present invention has discovered that, in the presence of the aforementioned solvents (e.g., 1,4-dioxane, water, and NMP) and at, for example, the aforementioned reaction temperatures, compounds of formula II can be completely converted. In some embodiments, the substitution reaction can take 10 to 30 hours.

[0190] In the above reaction, a mixture of the compound of formula II and the compound of formula SM3 is heated and refluxed to carry out the substitution reaction and complete conversion.

[0191] In some embodiments, purification can be performed after the reaction is complete. In some embodiments, the purification step includes: lowering the reaction temperature to 45 to 55°C, adding water (the amount of water may be 20 to 30V of the compound of formula (II), stirring the mixture at room temperature for 3 to 7 hours, and filtering to obtain the compound of formula (I). Alternatively, in some embodiments, in the purification step, after stirring at room temperature for 3 to 7 hours and filtering, the filter cake is washed with water, and the wet product is dissolved in DMSO (the amount may be 35 to 45V of the compound of formula II, for example 35.5V) at 85 to 95°C, then cooled to 45 to 55°C, adding methanol (the amount may be 31.5 to 40.5V of the compound of formula II, for example 34 to 37V), stirring the mixture at room temperature for 2 to 5 hours, filtering, and washing the filter cake with methanol to obtain the compound of formula (I). In some embodiments, the volume ratio of DMSO to methanol used in the cooling step is (2 to 1):1. In some embodiments, the filter cake is washed with methanol and then vacuum dried under reduced pressure to obtain the purified compound of formula (I).

[0192] In some embodiments, step (6) includes adding 1,4-dioxane, water, N,N-diisopropylethylamine, the hydrochloride of the compound of formula SM3, potassium iodide, the compound of formula II, and NMP to a reactor, heating and refluxing until the reaction is complete, lowering the temperature of the reaction solution to 50 ± 5 °C, adding water to the reaction solution, and further lowering the temperature to room temperature. After stirring at room temperature for 3 to 7 hours, filtering is performed, and the filter cake is washed with water. The wet product is dissolved in DMSO at 90 ± 5 °C, cooled to 50 ± 5 °C, methanol is added, and cooling is continued to room temperature. The mixture is stirred for 2 to 5 hours, filtered, the filter cake is washed with methanol, and the wet product is dried under reduced pressure at 50 ± 5 °C to obtain the compound of formula (I). In some embodiments, the amount and ratio of each substance in the reaction system, and the amount of each purified substance used in the purification step, are as described in any of the foregoing embodiments.

[0193] The compound of formula III is reacted with an acylation reagent to obtain the compound of formula II through the following steps (5):

[0194]

[0195] In step (5) above, a suitable acylation agent is selected from one or more of oxalyl chloride, thionyl chloride, trimethylchlorosilane, phosphorus oxychloride, and sulfonyl chloride, such as thionyl chloride. In some embodiments, the molar ratio of the acylation agent to the compound of formula III is 2.5 to 3.5. In some embodiments, the molar ratio is 2.8 to 3.1.

[0196] In step (5) above, a suitable organic solvent is selected from one or more of dichloromethane, acetonitrile, and tetrahydrofuran. In some embodiments, the organic solvent is dichloromethane.

[0197] In some embodiments, an activator (such as N,N-dimethylformamide) is added to the reactants in step (5). In some embodiments, the molar ratio of the activator to the compound of formula III is 1.8 to 2.3.

[0198] In step (5), the reaction temperature is 5°C to 15°C. The reaction time is 10 to 30 hours. In some embodiments, the acylation agent is added at -5°C to 5°C, and the mixture is stirred for 30 minutes to 5 hours, then heated to 5°C to 15°C for the reaction.

[0199] After the reaction is complete, the reaction is quenched with water, the mixture is filtered, and the filter cake is washed successively with dichloromethane and water to obtain the compound of formula II. In some embodiments, after the reaction is complete, 1 to 5 V of water is added to the reaction solution, the mixture is stirred for 6 to 14 hours, filtered, and the filter cake is washed successively with dichloromethane and water to obtain the compound of formula II. In this reaction, column chromatography is no longer used for post-treatment, thus improving the yield.

[0200] In some embodiments, step (5) includes adding the compound of formula III, dichloromethane and N,N-dimethylformamide to a reactor, adding thionyl chloride dropwise at -5°C to 5°C and stirring for 45 minutes to 3 hours, heating to 10±5°C and stirring until the reaction is complete, adding water, stirring for 8 to 12 hours, filtering, washing the filter cake sequentially with dichloromethane and water, and drying the wet product under reduced pressure at 50±5°C to obtain the compound of formula II.

[0201] According to this disclosure, a compound of formula IV is reacted with an organotin compound in the presence of a palladium catalyst to obtain a compound of formula III by the following step (4):

[0202]

[0203] In step (4) above, the solvent is selected from one or more of 1,4-dioxane, tetrahydrofuran, acetonitrile, DMF, dimethoxyethane, and diethylene glycol dimethyl ether; the organotin is tributyltin methanol; and the catalyst is selected from one or more of the following: Xphos Pd G2, Ruphos Pd G2, Brettphos Pd G1, and Xantphos Pd G3.

[0204] In some embodiments, step (4) includes reacting the compound of formula IV with an organotin reagent at 75 to 85°C for 12 to 30 hours in the presence of an organic solvent and a catalyst, adding an organic solvent and mercaptosilica gel, stirring at 75 to 85°C for 3 to 8 hours, filtering the mixture, adding mercaptosilica gel to the filtrate, stirring at 75 to 85°C for 3 to 10 hours, filtering the mixture, concentrating the filtrate, adding methyl tert-butyl ether, cooling to room temperature and stirring the mixture for 5 to 20 hours, and filtering to obtain the compound of formula III. The present invention has found that when crystallization is performed using 1,4-dioxane and methyl tert-butyl ether, the purity of the compound of formula III can reach 97%, and the mother liquor loss can be controlled to about 5%. In the above reaction, the volume ratio of 1,4-dioxane to methyl tert-butyl ether is (2.5 to 3.5):1.

[0205] In some embodiments, the molar ratio of the compound of formula IV to the organotin reagent is 1:(1 to 1.2).

[0206] According to this disclosure, the compound of formula V is reacted in an organic solvent under alkaline conditions to obtain the compound of formula IV by the following step (3):

[0207]

[0208] Alkaline conditions can be provided using inorganic or organic bases; for example, the inorganic base can be K2CO3, Na2CO3, K3PO4, or NaOH, such as K2CO3; the organic base is selected from one or more of Et3N (triethylamine), pyridine, DBU (1,8-diazabicycloundecane-7-ene), TMG (tetramethylguanidine), DABCO (triethylenediamine), and DIEA (N,N-diisopropylethylamine).

[0209] In some embodiments, the molar ratio of the inorganic base to the compound of formula V is (8 to 10):1.

[0210] In the above reaction, the organic solvent may be selected from one or more of THF, dioxane, IPA, CH3CN, DMSO, or NMP. In some embodiments, the organic solvent is NMP.

[0211] In the above reaction, the compound of formula V is added in batches, for example, in 2 to 5 batches, with each addition spaced 3 to 6 hours apart. Before addition, the temperature of the reaction solution can be lowered to 45 to 55°C, and after addition, the temperature is raised to the reaction temperature.

[0212] In the above reaction, the reaction temperature can be 115 to 125°C, and the reaction time can be 3 to 30 hours.

[0213] In the above reaction, after the reaction is complete, the temperature is lowered, water is added, the mixture is filtered, and the pH of the filtrate is adjusted with an acid (such as hydrochloric acid) to precipitate the compound of formula IV. In some embodiments, the temperature is lowered to room temperature, then water (containing 60% of the total amount of compound V) is added, and the mixture is stirred for 8 to 18 hours, then filtered to obtain a filtrate. The pH of the filtrate can be adjusted to 5 to 6, then stirred and filtered to obtain the compound of formula IV.

[0214] In some embodiments, step (3) includes: placing the compound of formula V into a reaction vessel, adding potassium carbonate and NMP, and stirring at 115 to 125°C for 3 to 6 hours. Cooling the temperature to 45 to 55°C, then placing the compound of formula V into the reaction vessel, raising the temperature to 115 to 125°C, and stirring for 3 to 6 hours, then cooling the temperature to 45 to 55°C, then adding the compound of formula V into the reaction vessel again, raising the temperature to 115 to 125°C, and stirring the mixture for 10 to 16 hours. Lowering the temperature to room temperature, then adding water, stirring the mixture for 8 to 18 hours, and filtering to obtain a filtrate. Adjusting the pH of the filtrate to 5 to 6, then stirring and filtering to obtain the compound of formula IV.

[0215] According to this disclosure, in the presence of a base and a coupling agent, a compound of formula VI and a compound of formula SM2 are reacted in an organic solvent to obtain a compound of formula V through the following step (2):

[0216]

[0217] In the above reaction, the base may be DIEA (N,N-diisopropylethylamine), DBU, or K₂CO₃. In some embodiments, the base is DIEA. The coupling agent may be HATU, HBTU, HOBT, and EDCI or CDI. In some embodiments, the coupling agent is CDI. The organic solvent may be CH₃CN or DMAc. In some embodiments, the compound of formula VI and the compound of formula SM₂ are reacted in CH₃CN with the participation of DIEA and CDI to obtain the compound of formula V.

[0218] The reaction temperature can be 75 to 85°C, and the reaction time can be 10 to 30 hours.

[0219] In some embodiments, the molar ratio of the compound of formula SM2 to the compound of formula VI is 2:3. After the reaction is complete, the temperature is lowered to room temperature, water is added to the reaction solution, the mixture is stirred for 10 to 20 hours, and filtered to obtain the compound of formula V. If necessary, the filter cake can be washed with water and dried under vacuum at 45 to 55°C to obtain the dried compound of formula V.

[0220] According to this disclosure, formula SM1 is reduced to obtain compound VI by the following step (1):

[0221]

[0222] The compound of formula SM1 and a platinum catalyst are reacted in an organic solvent under a certain hydrogen pressure to obtain the compound of formula VI. The hydrogen pressure can be from 0.27 to 0.3 MPa (inclusive). In some embodiments, the hydrogen pressure is 0.3 MPa. The platinum catalyst can be Pt-V / C. The organic solvent can be THF (tetrahydrofuran). In some embodiments, the weight ratio of the platinum catalyst (such as Pt-V / C) to the compound of formula SM1 is about 0.05 to 0.20.

[0223] After the reaction was complete, the temperature was lowered, the mixture was filtered through diatomaceous earth, washed with THF, and the filtrate was concentrated under reduced pressure, replaced twice with ACN, and concentrated to obtain an acetonitrile solution of the compound of formula VI.

[0224] The present invention also includes methods for preparing compounds of formulas I, II, III, IV, V and VI as described in the foregoing sections.

[0225] The present invention also includes the compounds of formulas I, II, III, IV, V and VI of this application themselves.

[0226] The present invention also includes the use of compounds SM1, SM2, SM3, I, II, III, IV, V, VI of this application for the preparation of compounds of formula (I).

[0227] The present invention also includes products containing compounds of formulas II, III, IV, V, and VI obtained in the above preparation steps, including but not limited to various solutions, filtrates, filter cakes, etc.

[0228] In some embodiments, this disclosure also provides a method for preparing a crystal form of a compound of formula (I), the method comprising: recrystallizing the compound of formula (I) in a solvent to obtain a crystal form, wherein the solvent is a mixture of acetic acid and methanol.

[0229] In the above method, compound (I) is dissolved in acetic acid, methanol is added dropwise, seed crystals are added, the mixture is stirred, and the solution is filtered to obtain crystal form A of compound (I). Compound (I) and acetic acid can be mixed at a temperature of 35 to 45°C.

[0230] In the above method, the amount of acetic acid is sufficient to completely dissolve the compound of formula (I). In some embodiments, the volume ratio of acetic acid to the compound of formula (I) can be (4 to 6):1. The volume ratio of acetic acid to methanol can be 1:(2.5-3.5), for example 1:3. The amount of seed crystals added can be from 0.2% to 2.5% by weight of the compound of formula (I).

[0231] In some embodiments, the compound of formula (I) is dissolved in acetic acid, followed by the dropwise addition of methanol, the addition of seed crystals, and stirring for 1 to 3 hours, followed by the dropwise addition of methanol. Stirring continues for 2 to 4 hours, the mixture is cooled to room temperature, and stirred for 4 to 10 hours. After filtration, crystal form A is obtained. The volume ratio of methanol added in the two additions can be 1:(2 to 4).

[0232] In some embodiments, the crystallization method includes adding acetic acid and the compound of formula (I) to a reactor, dissolving them at 35 to 45°C, then adding methanol dropwise, then adding seed crystals, stirring the mixture for 1 to 3 hours, then adding methanol dropwise, continuing to stir the mixture for 2 to 3 hours, then lowering the temperature to room temperature, then continuing to stir the mixture for 4 to 10 hours, filtering, washing with methanol, and drying to obtain the crystal form of the compound of formula (I).

[0233] In some embodiments, the crystallization method further includes the step of preparing a compound of formula (I) using any of the methods described herein.

[0234] Preparation of crystal form B

[0235] This document provides a method for preparing the disclosed crystal form B of compound (I), including exemplary reagents, quantities, and proportions. For example, crystal form B can be prepared by a method comprising the following steps:

[0236] (a) Providing a compound of formula (I) in solid form (e.g., any solid form disclosed herein);

[0237] (b) Optionally wash the compound of formula (I);

[0238] (c) Dissolve the compound of formula (I) in an acid, optionally wherein the acid is hydrochloric acid, 1.0N hydrochloric acid, sulfuric acid, an aqueous solution of sulfuric acid, or an aqueous solution of 0.25M sulfuric acid;

[0239] (d) Optionally wash the solution with an organic solvent, wherein the organic solvent is dichloromethane, and / or extract the solution, wherein the extraction solvent is ethyl acetate;

[0240] (e) Precipitate the compound of formula (I) by combining the solution with a base, wherein the base is, optionally, sodium bicarbonate in solid or solution form, sodium carbonate in solid or solution form, ammonia or ammonia solution;

[0241] (f) Optionally, the precipitate is dried, wherein the drying may include, for example, drying under an infrared lamp for up to 16 hours.

[0242] Crystal form B can be prepared by a method including the following steps:

[0243] 1. Optionally, the compound of formula (I) is washed with, for example, dichloromethane, methanol and ethyl acetate (optionally in this order), the compound of formula (I) being amorphous or crystalline or a mixture thereof; during each wash, the mixture is optionally stirred or mixed to expose the solid compound to the washing solvent before removing the solvent;

[0244] 2. Collect the solid substance and add acid, such as 1.0N hydrochloric acid, until dissolved (e.g., approximately 1 mL acid / 26.5 mg of free base of compound (I)).

[0245] 3. Extraction, for example, using approximately four times the amount of ethyl acetate (v / v) of the acid for three extractions.

[0246] 4. Add saturated sodium bicarbonate (NaHCO3) solution dropwise while stirring to precipitate crystal form B;

[0247] 5. Optionally, the solid material can be separated by filtration.

[0248] Crystal form B can be prepared by a method including the following steps:

[0249] 1. Dissolve the compound of formula (I) (which may be amorphous, crystalline, or a mixture thereof) in an acid (e.g., 1.0N hydrochloric acid) until dissolved (e.g., about 1 mL of acid / 45 to 50 mg of free base of compound (I)).

[0250] 2. Add sodium carbonate (Na2CO3) aqueous solution dropwise with stirring until a precipitate forms (approximately 1 mL Na2CO3 / 90 to 95 mg of compound (I));

[0251] 3. Optionally, the solid material can be separated by filtration.

[0252] In some embodiments, the method includes, for example, dissolving the compound of formula (I) in sulfuric acid (e.g., an aqueous solution of sulfuric acid such as 0.25 M) at ambient temperatures (e.g., 15°C to 30°C, 18°C ​​to 28°C, 20°C to 25°C), washing with a dichloromethane solution, and adding the aqueous solution to ammonia (e.g., 1.5 M) to form a slurry. The slurry is stirred at ambient temperature for 1 hour, filtered, and dried, for example, under an infrared lamp to obtain the crystal form of the compound of formula (I).

[0253] In some embodiments, the method comprises dissolving the compound of formula (I) (e.g., 1 g) in sulfuric acid (e.g., an aqueous solution of H₂SO₄) (e.g., 0.25 M, 10 mL). The resulting clear solution is combined with ammonia (e.g., 1.5 M, 5 mL) at ambient temperature. The resulting suspension is stirred, for example, for about 1 hour, filtered, washed with H₂O (e.g., 5 mL), and dried under an infrared lamp to obtain the crystal form of the third aspect disclosed herein. The material may be dried for 16 hours. The resulting material has a water content of 7.3% by weight as determined by Karl Fischer. XRPD shows a spectrum consistent with the crystal form of the third aspect disclosed herein (one hydrate molecule contains one molecule of the compound of formula (I) and approximately two water molecules). If heated for 40 hours, the material transforms into the crystal form of the second aspect disclosed herein, with a water content of 1.5% by weight as assessed by XRPD and TGA, as determined by KF.

[0254] VI. Methods and Applications

[0255] The compounds disclosed herein are selective inhibitors of PARP1, meaning that the compounds of this invention exhibit a stronger or more significant inhibitory effect on PARP1 than on PARP2 (or other PARPs). Therefore, the crystal forms of the compounds disclosed herein can be used to manufacture medicaments for treating or preventing clinical conditions that respond to inhibition of PARP1 activity.

[0256] As used in this article, a clinical condition that responds to the inhibition of PARP1 activity refers to a disease or condition whose onset or development can be treated or prevented by inhibiting PARP1 activity.

[0257] As used in this article, clinical conditions that respond to inhibition of PARP1 activity include cancer and other diseases that respond to inhibition of PARP1 activity, such as excessive cell death, including central nervous system diseases (such as stroke) and neurodegenerative diseases.

[0258] Cancers that respond to inhibition of PARP1 activity include, but are not limited to, liver cancer, melanoma, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, neuroblastoma, breast cancer, ovarian cancer, lung cancer (including, for example, small cell lung cancer), Wilms' tumor, cervical cancer, testicular cancer, soft tissue sarcoma, primary macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, and malignant melanoma. Tumors, gastric cancer, colon cancer, malignant pancreatic islet tumors, malignant carcinoid tumors, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, urogenital tumors, thyroid cancer, esophageal cancer, malignant hypercalcemia, cervical hyperplasia, renal cell carcinoma, endometrial cancer, polycythemia vera, idiopathic thrombocytosis, adrenocortical carcinoma, skin cancer, and prostate cancer.

[0259] Therefore, this disclosure provides the use of compounds of formula (I) in the preparation of medicaments for treating or preventing clinical conditions (such as cancers described herein) that respond to inhibition of PARP1 activity.

[0260] In some embodiments, this disclosure provides the use of a crystal form (e.g., crystal form A or crystal form B) of a compound of formula (I) according to this disclosure, or a mixture of two or more such crystal forms, in the preparation of a medicament for treating or preventing a clinical condition (such as cancer as described herein) that responds to inhibition of PARP1 activity.

[0261] In some embodiments, this disclosure provides the use of a crystalline form (e.g., crystalline form A or crystalline form B) of a compound of formula (I) according to this disclosure, or a mixture of two or more such crystalline forms, as a drug substance in the preparation of a medicament for treating or preventing a clinical condition (such as cancer as described herein) that responds to inhibition of PARP1 activity. The crystalline form or mixture of crystalline forms may further comprise an amorphous compound of formula (I).

[0262] The present invention also discloses a method for preparing a drug, the method comprising mixing a crystalline form (e.g., crystalline form A or crystalline form B) of a compound of formula (I) according to the present disclosure, or a mixture of two or more such crystalline forms, with a pharmaceutically acceptable carrier or excipient. The crystalline form or mixture of crystalline forms may further comprise an amorphous compound of formula (I).

[0263] Formulations of compounds of formula (I) may be pure crystal form A or pure crystal form B or pure amorphous material as disclosed herein, or may include mixtures of two or more of these.

[0264] The following examples further illustrate the embodiments described herein, but these examples should not be construed as limiting the scope of the embodiments described herein. Compounds, starting materials, and reagents useful in the methods described herein, such as compounds of formulas SM1, SM2, and SM3, can be obtained from commercial sources or prepared using methods known to those skilled in the art.

[0265] Example 1: Polymorph Screening and Suspension Competition Experiment

[0266] The preparation method of the starting material of the compound of formula (I) used for polymorph screening is as follows: Approximately 6 g of the compound of formula (I) was dissolved in DMSO (100 mL), stirred at 80 °C for 1 hour, cooled to room temperature, and poured into ice water (1 L), resulting in solid precipitation. The solid was collected by filtration, slurried with H₂O (100 mL) at 100 °C for 1 hour, and then filtered. The filtered solid was freeze-dried to obtain the starting material. X-ray powder diffraction (XRPD) results showed that the starting material was crystalline and named crystal form A, as shown below. Figure 20 As shown.

[0267] Using the aforementioned starting materials, polymorph screening experiments were conducted under 100 different conditions. These experiments employed methods including slow evaporation, gas-solid permeation, gas-liquid permeation, cyclic heating and cooling, suspension stirring (room temperature and 50°C), slow cooling, antisolvent addition, and grinding. Based on the characterization results from X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC), no new solid forms were obtained.

[0268] Another crystalline form of compound I was obtained by the following method: A sample of compound I was dissolved in a 1N HCl aqueous solution, and the pH was adjusted to 1 to 2. The solution was extracted with EtOAc. The aqueous phase was collected and the pH of the aqueous phase was adjusted to 7 to 9 with a NaHCO3 aqueous solution, and a solid precipitated. The mixture was filtered and the filter cake was dried to obtain the product. It exhibits XRPD diffraction peaks that are substantially consistent with crystalline form A, but additional diffraction peaks are present. These additional diffraction peaks disappear after heating the sample to 170°C and cooling it to room temperature.

[0269] A suspension competition experiment was conducted on crystal form A and another crystal form mentioned above to investigate their interconversion relationships under different temperature / solvent, room temperature, and water-active conditions. The test procedure was as follows: Approximately 5 mg of compound I, crystal form A, was weighed into each solvent, stirred and equilibrated at room temperature / 50°C for 2 hours, and then filtered. The filtrate was transferred to vials containing compound I, crystal form A, and the other crystal forms were further suspended and stirred at room temperature / 50°C, followed by XRPD testing. The different conditions and results of the suspension competition experiment are shown in Table 1.

[0270] Table 1: Results of the suspension competition experiment

[0271]

[0272]

[0273] The results showed that all obtained samples were of crystal form A.

[0274] XRPD uses the parameters shown in the table below for testing:

[0275]

[0276] Example 2: Solubility of crystal form A

[0277] The solubility of crystal form A in different single solvent systems was tested at 25℃ and 50℃. The results are shown in Table 2.

[0278] Table 2: Solubility of Crystal Form A in a Single Solvent System

[0279]

[0280]

[0281] The solubility of crystal form A in different binary solvent systems was tested at 25℃ and 50℃. The results are shown in Table 3.

[0282] Table 3: Solubility of Crystal Form A in Binary Solvent Systems

[0283]

[0284]

[0285] The results showed that, except in acetic acid and hexafluoroisopropanol, crystal form A had relatively low solubility in most of the tested single and binary solvent systems.

[0286] Example 3: Crystallization method using hexafluoroisopropanol / other solvents

[0287] Crystal form A exhibits high solubility in hexafluoroisopropanol (greater than 200 mg / mL at 25°C) (as shown in Example 2). Based on this result, crystal form A was recrystallized using hexafluoroisopropanol as the solvent and methyl tert-butyl ether, water, and methanol as the antisolvents, respectively. The recrystallization experiment of the hexafluoroisopropanol / water system proceeded as follows: Crystal form A was dissolved in hexafluoroisopropanol, and the solution was stirred at 30°C. Water and seed crystals were added, and the mixture was stirred. Additional water was added, and the mixture was continuously stirred. The mixture was then cooled to 10°C, stirred, filtered, washed, and dried to obtain the product.

[0288] The recrystallization procedures for the hexafluoroisopropanol / methyl tert-butyl ether and hexafluoroisopropanol / methanol systems were similar to those for the hexafluoroisopropanol / water system. The experimental results are shown in Table 4 below.

[0289] Table 4: Crystallization results of hexafluoroisopropanol / other solvent systems

[0290]

[0291] XRPD analysis of the product obtained from crystallization of the hexafluoroisopropanol / water system showed that crystal form A was obtained, but the XRPD contained additional diffraction peaks. The product crystallized from the hexafluoroisopropanol / methyl tert-butyl ether system was crystal form A, but the product contained a high amount of residual methyl tert-butyl ether. The product crystallized from the hexafluoroisopropanol / methanol system was also crystal form A, but the product contained a high amount of residual hexafluoroisopropanol.

[0292] Example 4: Crystallization method using acetic acid / other solvents

[0293] Crystal form A exhibits high solubility in acetic acid (greater than 200 mg / mL at 25°C) (as shown in Example 2). Based on this result, crystal form A was recrystallized using acetic acid as the solvent and methyl tert-butyl ether and methanol as the antisolvents, respectively. The recrystallization procedures for the acetic acid / methyl tert-butyl ether system and the acetic acid / methanol system were similar to those described in Example 3. The experimental results are shown in Table 5 below.

[0294] Table 5: Crystallization results of acetic acid / other solvent systems

[0295]

[0296] The products obtained from crystallization in both the acetic acid / methyl tert-butyl ether system and the acetic acid / methanol system are of crystal form A, and the content of each residual solvent in the products is extremely low.

[0297] Example 5: Preparation of crystal form A of compound of formula I

[0298] Step 1: Preparation of 5-bromo-2,3-difluoroaniline (Compound VI)

[0299]

[0300] 5-Bromo-2,3-difluoronitrobenzene (compound SM1, 14.8 kg, 62.2 mol), tetrahydrofuran (195.5 kg, 15V), and Pt-V / C (1.47 kg) were added to a hydrogenation reactor. The system was purged three times with hydrogen, maintained at 0.3 MPa, and stirred at 50°C for 20 hours. After cooling to room temperature, the mixture was filtered through diatomaceous earth and washed with tetrahydrofuran. The filtrate was concentrated under reduced pressure, then purged twice with acetonitrile, and finally concentrated under reduced pressure to obtain 32.6 kg of an acetonitrile solution of 5-bromo-2,3-difluoroaniline (compound VI, brown liquid, purity 98.2%, yield 94%).

[0301] HPLC(std):6.75min; MS(ESI):m / z=207.96[M+H] + ; 1 ¹H NMR (400MHz, DMSO-d⁶): 6.77

[0302] (d,J=6.8Hz,1H),6.70–6.66(m,1H),5.66(s,2H).

[0303] Step 2: Preparation of N-(5-bromo-2,3-difluorophenyl)-1H-pyrazole-5-carboxamide (compound V)

[0304]

[0305] In a reaction vessel, 1H-pyrazole-5-carboxylic acid (compound of formula SM2, 16.14 kg, 144.5 mol), N,N'-carbonyldiimidazole (18.68 kg, 115.2 mol), and acetonitrile (76 kg, 8.0 V) were added, and the mixture was stirred at 25 °C for 5 hours. Then, 5-bromo-2,3-difluoroaniline (compound of formula VI, 12.0 kg, 57.7 mol) and N,N-diisopropylethylamine (14.88 kg, 115.1 mol) were added, and the mixture was stirred at 80 °C for 18 hours. After cooling, water (300 kg, 25.0 V) was added and stirred at room temperature for 16 hours. The mixture was then filtered and washed with process water. The filter cake was dried under vacuum at 50 °C for 24 hours to obtain N-(5-bromo-2,3-difluorophenyl)-1H-pyrazole-5-carboxamide (compound V, 15.5 kg, white solid, purity 99.7%, yield 89%).

[0306] HPLC(std):10.23min; MS(ESI):m / z=301.97[M+H] + ; 1 H NMR (400MHz, DMSO-d6):

[0307] 13.55 (s, 1H), 9.95 (s, 1H), 7.94 (d, J = 2Hz, 1H), 7.88-7.87 (m, 1H), 7.66-7.63 (m, 1H), 6.81 (d, J = 2Hz 1H).

[0308] Step 3: Preparation of 7-bromo-9-flupyrazolo[1,5-a]quinoxaline-4(5H)-one (compound of formula IV)

[0309]

[0310] N-(5-bromo-2,3-difluorophenyl)-1H-pyrazole-5-carboxamide (compound V, 5.2 kg, 17.2 mol) was added to a reaction vessel, along with potassium carbonate (22 kg, 157 mol) and NMP (491 kg, 31V). The mixture was stirred at 120°C for 5 hours. The temperature was then lowered to 50°C, and N-(5-bromo-2,3-difluorophenyl)-1H-pyrazole-5-carboxamide (compound V, 5.2 kg, 17.2 mol) was added to the reaction vessel. The mixture was heated to 120°C and stirred for 5 hours. The temperature was then lowered to 50°C, and N-(5-bromo-2,3-difluorophenyl)-1H-pyrazole-5-carboxamide (compound V, 5.1 kg, 16.9 mol) was added to the reaction vessel. The mixture was then heated to 120°C and stirred for 13 hours. The mixture was cooled to room temperature, and process water (929 kg, 60.0 V) was added and stirred for 13 hours. The mixture was then filtered and the filtrate was collected. The pH of the filtrate was adjusted to 5-6 at room temperature with concentrated hydrochloric acid (28 kg, 1.5 V), stirred, filtered, and the wet product was dried under reduced pressure at 60 °C for 44 hours to obtain 7-bromo-9-fluoropyrazolo[1,5-a]quinoline-4(5H)-one (compound IV, 12.38 kg, white solid, purity 97.3%, yield 81%).

[0311] HPLC(std):6.74min; MS(ESI):m / z=281.97[M+H] + ; 1 H NMR (400MHz, DMSO-d6):

[0312] 12.09 (s, 1H), 8.13 (d, J = 2Hz, 1H), 7.54-7.51 (dd, J = 2, 11.5Hz, 1H), 7.35 (t, J = 2Hz, 1H), 7.20 (d, J = 2Hz, 1H).

[0313] Step 4: Preparation of 9-fluoro-7-(hydroxymethyl)pyrazolo[1,5-a]quinoxaline-4(5H)-one (compound of formula III)

[0314]

[0315] 1,4-Dioxane (190 kg, 20.5 V), 7-bromo-9-fluoro-5H-pyrazolo[1,5-a]quinoline-4-one (formula IV, 9.0 kg, 31.9 mol), Xphos Pd G2 (1.09 kg, 1.60 mol), and tributyltin methanol (12 kg, 36.4 mol) were added to a reaction vessel and stirred at 80 °C for 16 hours. 1,4-Dioxane (285 kg, 30.5 V) and mercaptosilica gel (4.6 kg) were added, and stirring continued at 80 °C for 5 hours. The mixture was filtered and washed with 1,4-Dioxane. Mercaptosilica gel (2.7 kg) was added to the filtrate, and stirring continued at 80 °C for 8 hours. The mixture was then filtered and washed with 1,4-Dioxane. The filtrate was concentrated under reduced pressure and stirred at 80°C for 3 hours. Methyl tert-butyl ether (111 kg, 16.5 V) was added at 50°C. The mixture was cooled to room temperature and stirred for 13 hours. The mixture was filtered, and the filter cake was washed with methyl tert-butyl ether. The wet product was dried at 50°C for 20 hours to obtain 9-fluoro-7-hydroxymethyl-5H-pyrazolo[1,5-a]quinoline-4-one (compound III, 7.34 kg, off-white solid, purity 96%, yield 80%).

[0316] HPLC(std):4.82min; MS(ESI):m / z=234.07[M+H] + ; 1 H NMR (400MHz, DMSO-d6):

[0317] 12.07 (s, 1H), 8.10 (d, J = 2Hz, 1H), 7.21 (s, 1H), 7.17 (d, J = 2Hz, 1H), 7.13-7.10 (d, J = 13.2Hz, 1H), 5.49 (s, 1H), 4.55 (s, 2H).

[0318] Step 5: Preparation of 7-(chloromethyl)-9-flupyrazolo[1,5-a]quinoxaline-4(5H)-one (compound of formula II)

[0319]

[0320] 9-fluoro-7-hydroxymethyl-5H-pyrazolo[1,5-a]quinoline-4-one (compound III, 5.73 kg, 24.6 mol), dichloromethane (220 kg, 29 V), and N,N-dimethylformamide (2.8 kg, 49.4 mol) were added to a reaction vessel. Thionyl chloride (15 kg, 72.1 mol) was added dropwise at 0 °C and stirred for 1 hour. The mixture was then heated to 10 °C and stirred for 18 hours. Process water (11 kg, 2 V) was added, and the mixture was stirred for 10 hours. The mixture was filtered, and the filter cake was washed successively with dichloromethane and process water. The wet product was dried under reduced pressure at 50 °C for 20 hours to obtain 7-chloromethyl-9-fluoro-5H-pyrazolo[1,5-a]quinoline-4-one (compound II, 6.3 kg, white solid, purity 96.0%, yield 91%).

[0321] HPLC(std):7.64min; MS(ESI):m / z=252.03[M+H] + ; 1 H NMR (400MHz, DMSO-d6):

[0322] 12.12(s,1H),8.12(d,J=1.76Hz,1H),7.32-7.29(d,J=12.5Hz,1H),7.28(s,1H),7.19(d,J=1.76Hz,1H),4.84(s,2H).

[0323] Step 6: Preparation of 5-(4-((9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoxalo-7-yl)methyl)piperazin-1-yl)-N,6-dimethylpyridine amide (compound of formula I)

[0324]

[0325] Synthesis: 1,4-Dioxane (49 kg, 10.5 V), process water (14 kg, 3 V), N,N-diisopropylethylamine (11.6 kg, 90.1 mol), N,6-dimethyl-5-(piperazin-1-yl)pyridineamide hydrochloride (formula SM3 compound hydrochloride, 5.1 kg, 21.5 mmol), potassium iodide (0.6 kg, 3.5 mol), 7-chloromethyl-9-fluoro-5H-pyrazolo[1,5-a]quinoline-4-one (formula II compound, 4.5 kg, 17.8 mol), and NMP (47 kg, 10 V) were added to the reactor. The mixture was heated to reflux and stirred for 16 hours. The temperature was lowered to 50 °C, and process water (112 kg, 25 V) was added dropwise to the reaction mixture. The mixture was cooled to room temperature and stirred for 5 hours. The mixture was filtered and washed with process water. The wet product was dissolved in DMSO (176 kg, 35.5 V) at 90 °C, then cooled to 50 °C and methanol (126 kg, 35.5 V) was added dropwise. The mixture was further cooled to room temperature and stirred for 3 hours. After filtration, the filter cake was washed with methanol. The wet product was dried under reduced pressure at 50 °C for 20 hours to give 5-(4-(9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoline-7-ylmethyl)-piperazin-1-yl)-6-methylpyridin-2-carboxylic acid formamide (compound I, 5.8 kg, white solid, purity 99.9%, yield 72%).

[0326] Recrystallization: Add acetic acid (20.5 kg, 5V) and 5-(4-(9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoline-7-ylmethyl)-piperazin-1-yl)-6-methylpyridine-2-carboxylic acid formamide (compound of formula I, 4.0 kg, 8.9 mol) to the reactor and dissolve at 40°C. Add methanol (12.7 kg, 4V) dropwise, add seed crystals (80 g), stir for 1–3 hours, and then add methanol (35 kg, 11V) dropwise. Continue stirring for 2-3 hours, cool to room temperature and stir for 7 hours, filter, wash with methanol, and dry the wet product under reduced pressure at 50°C for 21 hours to obtain 5-(4-(9-fluoro-4-oxo-4,5-dihydropyrazolo[1,5-a]quinoline-7-ylmethyl)-piperazin-1-yl)-6-methylpyridine-2-carboxylic acid formamide (crystal form A of compound I, 3.44 kg, off-white solid, purity 100%, yield 86%).

[0327] HPLC(std):11.60min; MS(ESI):m / z=450.21[M+H] + ; 1 H NMR (400MHz, DMSO-d6):

[0328] 12.00(s,1H),8.42–8.39(q,J=5.2,4.4Hz,1H),8.10(d,J=2Hz,1H),7.80(d,J=8.4Hz,1H),7.49(d,J=8.4Hz,1 H),7.24(s,1H),7.20-7.17(m,2H),3.62(s,2H),2.97(b,4H),2.80(d,J=4.8Hz,3H),2.60(b,4H),2.50(s,3H).

[0329] Example 6: Characterization of crystal form A

[0330] The crystal form A prepared in Example 5 was characterized by XRPD, TGA and DSC.

[0331] 1. Instruments and Methods

[0332] 1) X-ray powder diffractometer (XRPD)

[0333] XRPD testing was performed using a Bruker X-ray powder diffractometer, and the test parameters used are shown in the table below.

[0334]

[0335] 2) Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

[0336] TGA and DSC data were collected using a TA TA550 thermogravimetric analyzer and a TA DSC250 differential scanning calorimeter, respectively. The test parameters used are as follows.

[0337]

[0338] 2. Characterization

[0339] Example of XRPD pattern of crystal form A Figure 1 As shown in the figure, the corresponding XRPD diffraction peaks are shown in Table 6. The XRPD peak positions can vary between ±0.2°2θ.

[0340] Table 6: XRPD data for crystal form A

[0341]

[0342]

[0343] TGA curve as follows Figure 2 As shown in the figure, the TGA thermogram of crystal form A is displayed.

[0344] DSC curve as follows Figure 3As shown in the figure, the DSC thermogram of crystal form A is displayed. The DSC thermogram shows an endothermic peak at 320.7℃ (initial temperature).

[0345] Example 7: Stability of Crystal Form A

[0346] The stability of crystal form A was evaluated after 10 and 30 days at high temperature (60°C). XRPD of the samples was measured using the method described above. The XRPD overlay plots of crystal form A for high-temperature stability evaluation are shown below. Figure 4 As shown in the figure. The results indicate that the XRPD of the sample did not change significantly after 10 and 30 days, suggesting that the sample remains in crystal form A.

[0347] The stability of crystal form A under high humidity (25°C / 92.5% RH) for 10 and 30 days was evaluated. XRPD of the samples was measured using the method described above. The XRPD overlay plots of crystal form A for high humidity stability evaluation are shown below. Figure 5 As shown in the figure. The results indicate that the XRPD of the sample did not change significantly after 10 and 30 days, suggesting that the sample remains in crystal form A.

[0348] The stability of crystal form A under strong light irradiation (1×ICH) was evaluated. XRPD of the samples was measured using the method described above. The XRPD overlay of crystal form A for strong light irradiation stability evaluation is shown below. Figure 6 As shown in the figure. The results indicate that the XRPD of the sample did not change significantly under strong light irradiation, suggesting that the sample remains in crystal form A.

[0349] The procedure is as follows:

[0350] Place 0.5 g of sample in a clean quartz petri dish. Prepare two parallel samples and place them in a light chamber. Expose the samples to light according to the current edition of the Chinese Pharmacopoeia. When the total energy reaches 1.2 million lux·h and 200 watts·h / square meter (1 ICH), remove the two samples from the light chamber for testing.

[0351] Two parallel controls were treated in the same manner. The control samples were prepared using the same procedure, with the samples completely wrapped in aluminum foil. As described above, they were exposed to light and then tested. These samples were referred to as “1×ICH-dark”.

[0352] The stability of crystal form A under accelerated conditions (40°C / 75% RH) for 1 month, 3 months, and 6 months was evaluated. XRPD of the samples was measured using the method described above. The XRPD overlay plots of crystal form A for accelerated stability evaluation are shown below. Figure 7 As shown in the figure. The results indicate that the XRPD of the sample did not change significantly after 1 month, 3 months and 6 months, which suggests that the sample is still in crystal form A.

[0353] The stability of crystal form A was evaluated after 1, 3, 6, and 9 months of long-term storage at 25°C / 60% RH. The XRPD of the samples was measured using the method described above. The XRPD overlay plots of crystal form A for long-term stability evaluation are shown below. Figure 8 As shown in the figure. The results indicate that the XRPD of the sample did not change significantly after 1 month, 3 months, 6 months and 9 months, which suggests that the sample is still in crystal form A.

[0354] Example 8: Preparation of amorphous compound of formula (I)

[0355] The amorphous compound of formula (I) was prepared as follows: 9.0 g of compound (I) was dissolved in 960 g of dichloromethane and 240 mL of methanol, and spray-dried on a Buchi B-290 advanced spray dryer according to the parameters in Table 7. This spray drying produced an amorphous material, as evaluated by X-ray powder diffraction (XRPD) and polarized light microscopy (PLM), such as... Figure 18 and 19 As shown. For example, XRPD shows indistinguishable diffraction peaks, indicating that the material is amorphous.

[0356] Table 7

[0357]

[0358]

[0359] Example 9: First preparation of crystal form B of compound (I)

[0360] 105.6 mg of compound (I) was transferred to a porous glass filter and washed sequentially with 10 mL of dichloromethane (DCM), 5 mL of MeOH, and 10 mL of ethyl acetate (EtOAc). The solid was stirred with a glass rod to mix with each solvent and exposed to air before vacuum. After the addition of ethyl acetate, the mixture was stirred to form a suspension. The solid was collected and 1.0 N HCl was added until dissolved (approximately 4 mL). Ethyl acetate was added to the extract (3 × 15 mL), and the turbidity gradually decreased during washing. A saturated sodium bicarbonate solution was added dropwise with stirring to produce a precipitate.

[0361] The solids were separated by filtration and tested by XRPD, which showed that they were of crystal form B.

[0362] Example of XRPD pattern implementation for crystal form B Figure 9A As shown in the figure, the corresponding XRPD peak positions are listed in Table 8. The position of each listed XRPD peak can vary between ±0.2°2θ. Figure 9BAn example of the XRPD pattern of crystal form B after drying under an infrared lamp for 16 hours is shown.

[0363] Table 8: XRPD data for crystal form B

[0364]

[0365]

[0366] XRPD can be tested using the following parameters:

[0367] Table 9: XRPD Parameters

[0368]

[0369] Example 10: Second preparation of crystal form B of compound (I)

[0370] The preparation of crystalline form B of compound (I) was as follows: 93.7 mg of compound (I) was dissolved in approximately 2 mL of 1.0 N HCl. Saturated Na₂CO₃ was added dropwise with stirring, and 1 mL was added after the precipitate had completely formed. The stirring time was less than 5 minutes. The solid was separated by filtration and analyzed by XRPD, revealing that the solid was crystalline form B.

[0371] Crystal form B was characterized by TGA and DSC. TGA showed a rapid weight loss of at least 4.75% from ambient temperature, while DSC showed a large desolvation endothermic peak at 100.65 °C, and a distinct endothermic peak starting around 296 °C. Figure 10A ).

[0372] TGA data were collected using TA Instruments' TA Discovery 550TGA (CPNJ-E3). DSC was performed using TA Instruments' TA 2500DSC (CPNJ-E27). DSC was calibrated using indium reference standards, and TGA was calibrated using nickel reference standards. Detailed parameters used are listed in Table 10.

[0373] Table 10: Parameters measured by TGA and DSC

[0374]

[0375]

[0376] After drying the material with, for example, an infrared lamp for 16 hours, the same material was analyzed using a similar procedure. The results were as follows: Figure 10B As shown in (DSC) and 10C (TGA).

[0377] Example 11: Third preparation of crystal form B of compound (I)

[0378] The compound of formula (I) (1 g) was dissolved in an aqueous sulfuric acid solution (0.25 M, 10 mL) at ambient temperature, and the resulting solution was washed with dichloromethane (DCM, 10 mL). The aqueous solution was added to ammonia water (1.5 M, 5 mL) and stirred. The resulting solid-liquid mixture was stirred at ambient temperature for 1 hour, filtered, and dried under an infrared lamp to obtain the compound of formula (I) (approximately 0.95 g). Figure 9A An example of XRPD for crystal form B is shown. Figure 9B An example of the XRPD pattern of crystal form B after drying under an infrared lamp for 16 hours is shown.

[0379] Example 12: Single crystal cultivation and structural analysis of crystal form A

[0380] Anhydrous form: Single crystals of crystal form A were obtained by gas-liquid percolation in a solvent system of acetic acid (normal solvent) / 2-methyltetrahydrofuran (antisolvent). Single-crystal X-ray diffraction (SCXRD) characterization results showed that the single crystal of crystal form A belongs to the triclinic crystal system, space group P1. Its unit cell parameters are as follows: α=90.5230(10)°, β=90.508(2)°, γ=105.180(2)°, Single-crystal structure analysis shows that the asymmetric unit of this single-crystal structure consists of only one compound molecule of formula (I), such as Figure 11 As shown, this indicates that single crystals of crystal form A are anhydrous. A schematic diagram of molecular packing observed along the a-axis of the crystal is shown below. Figure 12 As shown, the crystal lattice contains small cavities that do not contain any water of crystallization molecules or other solvent molecules.

[0381] Hydrate: Single crystals of crystal form A were obtained by sequentially cultivating them through gas-liquid permeation and slow evaporation in a solvent system of dimethyl sulfoxide (positive solvent) / 2-methyltetrahydrofuran (antisolvent). SCXRD characterization results showed that the single crystals of crystal form A belong to the triclinic crystal system, space group P1. Its unit cell parameters are as follows: α=90.4750(10)°, β=90.2170(10)°, γ=104.760(2)°, Single-crystal structure analysis shows that the asymmetric unit of this single-crystal structure consists of a compound molecule of formula I and a water of crystallization molecule with a chemical occupancy of approximately 0.2 located in the lattice cavity, such as... Figure 13 As shown, this indicates that the single crystal of crystal form A is a non-stoichiometric hydrate (the molar ratio of free base molecules to water of crystallization molecules in the single crystal structure is 1:0.2). A schematic diagram of molecular packing observed along the a-axis of the crystal is shown below. Figure 14 As shown.

[0382] The hydrated and anhydrous forms differ only slightly in structure, but these differences are reflected in a small shift in diffraction peaks. Figure 15 and Figure 16 The hydrate structure contains only 20% water, indicating the possible presence of a monohydrate, but the monohydrate structure was not successfully captured. The hydrogen bonding observed in the structure suggests that a monohydrate is feasible. The anhydrous structure is almost identical, with a volume difference of only 0.26%, and the orientation and packing are also almost identical. Figure 17 ).

[0383] Example 13: Chemiluminescence determination of PARP1 and PARP2

[0384] Diluted buffer (40 ng enzyme / well) containing recombinant poly(ADP-ribose) polymerase 1 and poly(ADP-ribose) polymerase 2 (PARP1 and PARP2) was added to 96-well plates coated with recombinant proteins along with the test compound. The plates were incubated at room temperature for 1 hour. Then, 50 μL of 0.3 ng / mL horseradish peroxidase with streptavidin-HRP was added to each well. The plates were incubated at room temperature for 30 minutes. Finally, the plates were treated with streptavidin-HRP, and then ELISA ECL substrate was added to produce chemiluminescence that could be measured using a chemiluminescence reader. The inhibition of PARP1 / 2 enzyme activity by the test compound was calculated using the following formula.

[0385]

[0386] IC 50 The values ​​were obtained by fitting the S-shaped dose-response curve equation using XL Fit software. The curve equation is Y = 100 / (1 + 10^(logC - logIC)). 50 )), where C is the concentration of the compound.

[0387] The results showed that the disclosed compounds exhibited inhibitory effects (IC50) on PARP1 and PARP2 enzymes. 50 The concentrations were 1.71 nM and 868 nM, respectively. Compared with PARP2, the compounds disclosed herein exhibit selective inhibitory activity against PARP1 enzyme activity.

[0388] Example 14: Inhibitory activity of compound (I) on the growth of BRCA-mutant human breast cancer MDA-MB-436 cells Measurement

[0389] The inhibitory activity of compound (I) on the growth of BRCA-mutant human breast cancer MDA-MB-436 cells was determined. MDA-MB-436 cells were cultured in complete medium (DMEM + 10% FBS + insulin + glutathione). When confluence reached approximately 80%, the cells were digested and gently removed from the bottom of the culture dish using a 1 mL pipette. The cell suspension was collected and centrifuged at 500 rpm for 3 minutes. The supernatant was discarded, and the cell pellet was resuspended in complete medium. Cells were seeded at an appropriate ratio in culture dishes and then cultured at 37°C in a 5% CO2 incubator. Measurements were performed when the cells were in optimal condition and confluence reached 80%. Cells in the logarithmic growth phase were centrifuged, and the culture supernatant was removed. Cells were resuspended in fresh complete medium and counted. The resuspended cells were seeded at 3000 cells / well in 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator. The compounds were prepared as follows: 5 μL of the 1000× compound solution was added to 120 μL of culture medium (25-fold dilution), and the 1000× diluted test compound solution was diluted to a 40× test compound solution. The solutions were mixed by shaking. 0.1% DMSO was used as a control.

[0390] On the second day, the 96-well plates inoculated with cells were removed from the incubator, and the culture supernatant was removed. Then, 195 μL / well of fresh culture medium and 5 μL / well of the 40× test compound solution were added to each well. Finally, the plates were incubated at 37°C in a 5% CO2 incubator for 7 days. On the fourth day, the culture medium containing the compound was replaced. After 7 days, 20 μL of CCK-8 was added to each well and gently shaken, then incubated for 4 hours. After incubation, the plates were shaken for 5 minutes. The absorbance values ​​at 450 nm or 650 nm wavelengths were recorded using a multimeter (OD = absorbance at 450 nm - absorbance at 650 nm).

[0391] Data were analyzed using GraphPad Prism 6.0 software. The inhibitory activity of the compound on cell proliferation was plotted using cell viability versus compound concentration as coordinates. Cell viability percentage % = (OD) / (Cell viability percentage % = 0.05%) ... 化合物 -OD 背景 ) / (OD DMSO -OD 背景 )×100. IC 50 The value was fitted using the sigmoid dose-response curve equation: Y = 100 / (1 + 10^(logC - logIC)) 50 )), where C is the concentration of the compound.

[0392] The results showed that the disclosed compound had a good growth inhibitory effect on MDA-MB-436 cells with BRCA mutations (IC50).50 (=1.20nM).

Claims

1. A crystal form of a compound of formula (I), The crystal form is characterized by X-ray powder diffraction (XRPD) spectra showing peaks at at least four of the following locations: 10.8±0.2°2θ、 13.2±0.2°2θ、 16.2±0.2°2θ、 20.2±0.2°2θ、 21.8±0.2°2θ, and 29.0±0.2°2θ.

2. The crystal form according to claim 1, wherein the crystal form is characterized by an XRPD spectrum showing peaks at the following locations: 10.8±0.2°2θ、 20.2±0.2°2θ、 21.8±0.2°2θ, and 29.0±0.2°2θ.

3. The crystal form according to claim 1, wherein the crystal form is characterized by an XRPD spectrum showing peaks at the following locations: 10.8±0.2°2θ、 13.2±0.2°2θ、 16.2±0.2°2θ、 20.2±0.2°2θ, and 21.8±0.2°2θ.

4. The crystal form according to claim 1, wherein the crystal form is characterized by an XRPD spectrum showing peaks at the following locations: 10.8±0.2°2θ、 13.2±0.2°2θ、 16.2±0.2°2θ, and 29.0±0.2°2θ.

5. The crystal form according to claim 1, wherein the crystal form is characterized by an XRPD spectrum showing peaks at the following locations: 10.8±0.2°2θ、 13.2±0.2°2θ、 16.2±0.2°2θ、 20.2±0.2°2θ、 21.8±0.2°2θ, and 29.0±0.2°2θ.

6. A crystal form of a compound of formula (I): in, The XRPD spectrum of the crystal form includes diffraction peaks at at least four of the following positions: 5.4±0.2°2θ、 9.9±0.2°2θ、 10.8±0.2°2θ、 11.4±0.2°2θ 13.2±0.2°2θ、 16.2±0.2°2θ、 17.7±0.2°2θ 20.2±0.2°2θ、 20.5±0.2°2θ、 22.6±0.2°2θ, and 24.3±0.2°2θ.

7. A crystal form of a compound of formula (I): The crystal form described therein has essentially the same XRPD spectrum as shown in Figure 1.

8. A crystal form of a compound of formula (I): The crystal form described therein has essentially the same thermogravimetric analysis (TGA) curve as shown in Figure 2.

9. A crystal form of a compound of formula (I): The crystal form described therein has a differential scanning calorimetry (DSC) curve that is substantially the same as that shown in Figure 3.

10. A crystal form of a compound of formula (I): The crystal form said to have at least one of the following: (a) The XRPD spectrum is essentially the same as that shown in Figure 1; (b) The TGA curve is essentially the same as that shown in Figure 2; and (c) The DSC curve is essentially the same as that shown in Figure 3.

11. A crystal form of a compound of formula (I): The crystal form described herein has a DSC curve showing a peak at 322.4℃±5℃.

12. A crystal form of a compound of formula (I), The crystal form is characterized by an XRPD spectrum showing peaks at at least four of the following locations: 4.6±0.2°2θ、 9.3±0.2°2θ、 10.8±0.2°2θ、 13.9±0.2°2θ、 21.3±0.2°2θ、 21.9±0.2°2θ、 23.5±0.2°2θ, and 24.8±0.2°2θ.

13. A crystal form of a compound of formula (I): The crystal form described therein has essentially the same XRPD spectrum as shown in Figure 9A or Figure 9B.

14. A crystal form of a compound of formula (I): The crystal form described therein has a TGA curve that is substantially the same as that shown in Figure 10A or Figure 10C.

15. A crystal form of a compound of formula (I): The crystal form described therein has a DSC curve that is substantially the same as that shown in Figure 10A or Figure 10B.

16. A crystal form of a compound of formula (I): The crystal form said to have at least one of the following: (a) XRPD is essentially the same as shown in Figure 9A or Figure 9B; (b) The TGA curve is substantially the same as that shown in Figure 10A or Figure 10C; and (c) The DSC curve is essentially the same as that shown in Figure 10A or Figure 10B.

17. A crystal form of a compound of formula (I): The crystal form has a DSC curve showing DSC endothermic peaks at any one, two, or all three of the following temperatures: 101℃±5℃, 288℃±5℃, and 309℃±5℃ and / or endothermic onset temperatures at any one, two, or all three of the following temperatures: 57℃±5℃, 280℃±5℃, and 295℃±5℃, or has a DSC curve showing DSC endothermic peaks at any one or two of the following temperatures: 83℃±5℃ and 318℃±5℃ and / or endothermic onset temperatures at any one or two of the following temperatures: 21℃±5℃ and 296℃±5℃.

18. A composition comprising the crystal form according to any one of claims 1 to 17 and a pharmaceutically acceptable carrier or excipient.

19. The composition of claim 18, wherein the composition further comprises at least one known anticancer drug or a pharmaceutically acceptable salt thereof.

20. The composition according to claim 19, wherein the at least one known anticancer drug is selected from the group consisting of: abiraterone, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, doxorubicin, dalorutamide, enzalutamide, epirubicin, azorubicin, mitoxantrone, methylhydroxyalotexin, etoposide, 5-azacitidine, gemcitabine, 5 -Fluorouracil, capecitabine, methotrexate, 5-fluoro-2'-deoxyuridine, fludarabine, nerabine, ara-C, pralatrexate, prednisone, pemetrexed, hydroxyurea, thioguanine, colchicine, vinblastine, vincristine, vinorelbine, paclitaxel, ixapril, cabazitaxel, docetaxel, panitumumab, nexituzumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, atorvastatin, olfamol, rituximab Anti-, alenmab, teimomab, tosimob, butuximab, daralimumab, elutuzumab, T-DM1, ditoximab, bonatumab, ipilimumab, bevacizumab, trastuzumab, rituximab, imatinib, gefitinib, erlotinib, ostinotinib, afatinib, ceritinib, alectinib, crizotinib, erlotinib, lapatinib, sorutinib, lafenamide, regorafenib, vemurafenib, dabrafenib, aflibercept, sunitinib, nilotinib, dasatinib, bosutinib Pralatinib, brutinib, cabozantinib, lenvatinib, vandetanib, trametinib, carbitinib, axitinib, tesiromoximide, ederalis, pazopanib, everolimus, tamoxifen, letrozole, fulvestrant, mitoramicin, octreotide, retinic acid, arsenic, zoledronic acid, bortezomib, carfilzomib, ixazomib, vemodega, solidega, denosumab, thalidomide, lenalidomide, venetumab, adefovir (recombinant human interleukin-2), ciproxetine-T (prostate cancer treatment vaccine).

21. A method for preparing a composition, the method comprising combining a crystal form according to any one of claims 1 to 17 with a pharmaceutically acceptable excipient or carrier.

22. A composition prepared by the method according to claim 21.

23. The composition of claim 22, wherein the composition further comprises at least one known anticancer drug or a pharmaceutically acceptable salt thereof.

24. The composition according to claim 23, wherein the at least one known anticancer drug is selected from the group consisting of: abiraterone, busulfan, melphalan, chlorambucil, cyclophosphamide, ifosfamide, temozolomide, bendamustine, cisplatin, mitomycin C, bleomycin, carboplatin, camptothecin, irinotecan, topotecan, doxorubicin, dalorutamide, enzalutamide, epirubicin, azorubicin, mitoxantrone, methylhydroxyalotexin, etoposide, 5-azacitidine, gemcitabine, 5 -Fluorouracil, capecitabine, methotrexate, 5-fluoro-2'-deoxyuridine, fludarabine, nerabine, ara-C, pralatrexate, prednisone, pemetrexed, hydroxyurea, thioguanine, colchicine, vinblastine, vincristine, vinorelbine, paclitaxel, ixapril, cabazitaxel, docetaxel, panitumumab, nexituzumab, nivolumab, pembrolizumab, ramucirumab, bevacizumab, pertuzumab, trastuzumab, cetuximab, atorvastatin, olfamol, rituximab Anti-, alenmab, teimomab, tosimob, butuximab, daralimumab, elutuzumab, T-DM1, ditoximab, bonatumab, ipilimumab, bevacizumab, trastuzumab, rituximab, imatinib, gefitinib, erlotinib, ostinotinib, afatinib, ceritinib, alectinib, crizotinib, erlotinib, lapatinib, sorutinib, lafenamide, regorafenib, vemurafenib, dabrafenib, aflibercept, sunitinib, nilotinib, dasatinib, bosutinib Pralatinib, brutinib, cabozantinib, lenvatinib, vandetanib, trametinib, carbitinib, axitinib, tesiromoximide, ederalis, pazopanib, everolimus, tamoxifen, letrozole, fulvestrant, mitoramicin, octreotide, retinic acid, arsenic, zoledronic acid, bortezomib, carfilzomib, ixazomib, vemodega, solidega, denosumab, thalidomide, lenalidomide, venetumab, adefovir (recombinant human interleukin-2), ciproxetine-T (prostate cancer treatment vaccine).

25. A method for preparing a compound of formula (I), wherein the method comprises the following steps: (a) Reduction of the (SM1) compound to obtain the (VI) compound: (b) Preparation of compound (V) from compounds of formula (VI) and formula (SM2) in an organic solvent in the presence of a base and a coupling agent: (c) Reacting compound (V) under alkaline conditions in an organic solvent to give compound (IV): (d) Reacting compound (IV) with organotin in the presence of a palladium catalyst to give compound (III): (e) Preparation of compound (II) from compound (III) and acylation reagent: (f) Preparation of compound (I) by substitution reaction between compound (II) and compound (SM3) in the presence of a base and a solvent:

26. A method for preparing a crystalline form of a compound of formula (I), wherein the method involves recrystallizing the compound of formula (I) in a mixture of acetic acid and methanol:

27. A method for preparing the crystal form of a compound of formula (I), The method includes the following steps: (a) Providing the compound of formula (I) in solid form; (b) Optionally wash the compound of formula (I); (c) Dissolve the compound of formula (I) in an acid, optionally wherein the acid is hydrochloric acid, 1.0N hydrochloric acid, sulfuric acid, an aqueous solution of sulfuric acid, or an aqueous solution of 0.25M sulfuric acid; (d) Optionally wash the solution with an organic solvent, wherein the organic solvent is dichloromethane, and / or extract the solution, wherein the extraction solvent is ethyl acetate; (e) Precipitate the compound of formula (I) by combining the solution with a base, wherein the base is, optionally, sodium bicarbonate in solid or solution form, sodium carbonate in solid or solution form, ammonia or ammonia solution; (f) Optionally, the precipitate is dried, wherein the drying is optionally performed under an infrared lamp for up to 16 hours.

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