Solid forms of bcl-2 inhibitors, method of preparation, and use thereof

Abstract

This invention relates to the Bcl-2 inhibitor 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide in solid form, particularly in crystalline form, pharmaceutical compositions comprising the solid form, a process for preparing the solid form, and a method of using the invention.

Description

[Technical Field]

[0001] This article discloses the solid form of the Bcl-2 inhibitor 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide, pharmaceutical compositions comprising the solid form, the process for preparing the solid form, and the method of using it. [Previous Technology]

[0002] Planned cell death, or apoptosis, occurs in multicellular organisms to process damaged or unwanted cells, which is essential for normal tissue homeostasis (Br. J. Cancer 1972, 26, 239). However, defective apoptotic processes have been involved in a wide variety of diseases. Excessive apoptosis leads to shrinkage, while insufficient apoptosis leads to uncontrolled cell proliferation, such as in cancer (Cell 2011, 144, 646). Apoptotic cell death is a hallmark of resistant cancers and promotes the development of chemoresistance (Nat Med. 2004, 10, 789-799). Several key pathways controlling apoptosis are often altered in cancer. Some factors, such as the Fas receptor and caspase, promote apoptosis, while some members of the B-cell lymphoma 2 (Bcl-2) protein family inhibit it. Negative regulation of apoptosis suppresses cell death signaling pathways, explaining why tumors evade cell death and develop drug resistance.

[0003] There are two distinct apoptosis pathways, including extrinsic and intrinsic pathways. The extrinsic pathway is activated in response to the binding of death-inducing ligands to cell surface death receptors (Nat Rev Drug Discov. [Nature Reviews Drug Discovery] 2017 16, 273-284). The B-cell lymphoma 2 (BCL-2) gene family is a group of proteins homologous to Bcl-2 proteins, encoding more than 20 proteins that regulate intrinsic apoptosis pathways. Bcl-2 family proteins are characterized by containing at least one of four conserved Bcl-2 homologous (BH) domains (BH1, BH2, BH3, and BH4) (Nat. Rev. Cancer [Nature Reviews Cancer] 2008, 8, 121; Mol. Cell [Molecular Cell] 2010, 37, 299; Nat. Rev. Mol. Cell Biol. [Nature Molecular Cell Biology Reviews] 2014, 15, 49). The Bcl-2 family of proteins consists of pro-apoptotic and anti-apoptotic molecules, and can be divided into three subfamilies based on sequence homology within the four BH domains: (1) a subfamily sharing sequence homology across all four BH domains, such as the anti-apoptotic Bcl-2, Bcl-XL, and Bcl-w; (2) a subfamily sharing sequence homology within BH1, BH2, and BH4, such as the pro-apoptotic Bax and Bak; and (3) a subfamily sharing sequence homology only within BH3, such as the pro-apoptotic Bik, Bid, and HRK. One of the unique characteristics of the Bcl-2 family of proteins is the heterodimerization between the anti-apoptotic and pro-apoptotic proteins, which is thought to inhibit the biological activity of their counterparts. This heterodimerization is mediated by inserting the BH3 region of the pro-apoptotic protein into a hydrophobic cleft composed of the BH1, BH2, and BH3 regions of the anti-apoptotic proteins. In addition to BH1 and BH2, the BH4 domain is also required for anti-apoptotic activity. In contrast, the BH3 domain is essential and sufficient to maintain pro-apoptotic activity.

[0004] Similar to oncogene addiction, where tumor cells depend on a single dominant gene for survival, tumor cells may also become dependent on Bcl-2 in order to survive. Bcl-2 overexpression is frequently found in acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), relapsed / refractory chronic lymphocytic leukemia (CLL), follicular lymphoma (FL), non-Hodgkin's lymphoma (NHL), and solid tumors (e.g., pancreatic cancer, prostate cancer, breast cancer, and small cell and non-small cell lung cancer) (Cancer 2001, 92, 1122-1129; Cancer Biol. 2003; 13:115-23; Curr. Cancer Drug Targets 2008, 8, 207-222; Cancer 2011, 3, 1527-1549). Dysregulation of the apoptosis pathway is associated with the pathology of the following diseases: other major diseases (upregulated apoptosis) (e.g., neurodegenerative diseases such as Alzheimer's disease); and proliferative diseases (downregulated apoptosis), such as cancer, autoimmune diseases, and prothrombotic diseases.

[0005] International Publication WO 2019 / 210828 discloses a series of Bcl-2 inhibitors, particularly 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide (hereinafter referred to as Compound 1), which selectively inhibits Bcl-2 protein for the treatment of dysregulated apoptotic diseases such as cancer, autoimmune diseases and prothrombotic disorders.

[0006] Compound 1 has 13 rotatable bonds and a high molecular weight (Mw > 800). Molecules with high conformational flexibility are often extremely difficult to crystallize, and the most important molecular descriptors responsible for the crystallization behavior of such molecules are related to the number of rotatable bonds and the length of the alkyl side chain (Bruno C. Hancock. Predicting the Crystallization Propensity of Drug-Like Molecules. Journal of Pharmaceutical Sciences, 2017, 106: 28-30). In practice, for particular compounds, especially those with high molecular weights and many rotatable bonds, it is impossible to predict whether a pure physical form can be obtained and which physical forms will be stable and suitable for pharmaceutical use. Similarly, it is equally impossible to predict whether a particular crystalline solid form with the desired chemical and physical properties suitable for pharmaceutical formulations can be produced.

[0007] For all the reasons mentioned above, there is a great need to find a crystalline form of compound 1 that provides good stability and good manufacturability. This disclosure advantageously satisfies one or more of these requirements. [Summary of the Invention]

[0008] This disclosure addresses the aforementioned challenges and needs by providing a solid form, preferably a crystalline form, of Compound 1 suitable for pharmaceutical use. Although Compound 1 is found to have multiple freely rotating bonds and a high molecular weight exceeding 800, the inventors of this disclosure unexpectedly discovered twenty-one crystalline forms of Compound 1, including six anhydrous forms (forms B, S, U, M, F, and N), four hydrated / anhydrous forms (forms H, R, L, and T), and eleven solvates (forms A, C, D, E, G, I, J, K, O, P, and Q), wherein isomorphism occurs during the formation of form I, form L is a metastable form, forms N and T are interconvertible during storage, and form S is obtained by heating form R to 150°C.

[0009] The inventors of this invention have discovered that form A, an EtOAc solvate of compound 1, has good physical properties, including good physical stability and good solubility. However, it is difficult to control the ethyl acetate content of form A during manufacturing, storage, and formulation, and form A can be converted to form B after heating to 160°C, cooling back to room temperature, and re-exposing to an air atmosphere.

[0010] Solvent forms C, D, J, K and O and anhydrous form F can be converted into anhydrous form B after being heated to a high temperature; forms K and F can spontaneously convert into form B after long-term storage, and form R can be converted into anhydrous form S after being heated to 150°C.

[0011] The anhydrous forms B, S and M, compared with forms F, H, N and R, exhibit better physicochemical stability when exposed to 25°C / 60% RH and 40°C / 75% RH for 1 week and 80°C / sealed for 24 hours.

[0012] In addition, form B exhibits good thermodynamic stability, a high melting point, and slight hygroscopicity, with a water absorption rate of 0.9% at 25°C / 80% RH. It also shows good physicochemical and thermodynamic stability after exposure to 25°C / 80% RH and shaking in acetone / H₂O (1:9, v / v) and H₂O for about 4 days.

[0013] The inventors of this invention attempted to scale up form B, but were unable to obtain the desired crystal form directly using conventional crystallization methods. Furthermore, form B could only be obtained by heating form A at approximately 100°C or treating form K in certain solvents, which did not meet the requirements of the scale-up process. Using anhydrous CHCl3 and heptane as solvents, form M, which exhibited good stability, was obtained. However, CHCl3 is environmentally unfriendly and belongs to Group 2, with a low permissible daily exposure (PDE) of 0.6 mg / day according to ICH guidelines. Unexpectedly, replacing CHCl3 with DCM in the recrystallization step yielded an anhydrous form U of compound 1, which is also reproducible and suitable for scale-up processes. Form U exhibited good physicochemical, thermodynamic, and physical stability; for example, when stored for up to 6 months at 25 ± 2°C / 60 ± 5% RH or 40 ± 2°C / 75 ± 5% RH, there was no significant change in chemical purity, crystal form, or optical purity. In addition, only form U can effectively remove the key dimer impurity in manufacturing, namely the process impurity formed by the reaction of acidic intermediate (S)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(2-(2-(2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzoic acid with compound 1.

[0014] Although form U has a lower melting point than form B, form U does not have the problems of API and drug formulation preparation, scale-up processes, solvent residues, and identification, and it has good stability and solution crystallization ability. Therefore, form U is more suitable for manufacturing and drug formulation.

[0015] In a first aspect, this document discloses the crystalline form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide, designated as form A.

[0016] In a second aspect, this article discloses the crystalline form of compound 1, which is an EtOAc solvate containing about 1 mol of EtOAc per mol.

[0017] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 16.5 ± 0.1° and 24.5 ± 0.1°.

[0018] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 12.4 ± 0.1°, 16.5 ± 0.1° and 24.5 ± 0.1°.

[0019] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 12.4 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

[0020] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

[0021] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

[0022] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

[0023] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

[0024] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.7 ± 0.1°, and 24.5 ± 0.1°.

[0025] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.9 ± 0.1°, 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.7 ± 0.1°, and 24.5 ± 0.1°.

[0026] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has angular values ​​of 6.9 ± 0.1°, 7.4 ± 0.1°, 8.8 ± 0.1°, 10.6 ± 0.1°, 10.9 ± 0.1°, 12.4 ± 0.1°, 12.7 ± 0.1°, 13.1 ± 0.1°, 13.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 14.7 ± 0.1°, 14.9 ± 0.1°, 15.4 ± 0.1°, 16.2 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 17.5 ± 0.1°, 18.2 ± 0.1°, 18.5 ± 0.1°, and 19.1 ± 0.1°. Diffraction peaks are present at 0.1°, 19.5 ± 0.1°, 20.7 ± 0.1°, 21.1 ± 0.1°, 21.8 ± 0.1°, 22.4 ± 0.1°, 22.8 ± 0.1°, 23.3 ± 0.1°, 23.8 ± 0.1°, 24.1 ± 0.1°, 24.5 ± 0.1°, 25.8 ± 0.1°, 26.7 ± 0.1°, 27.1 ± 0.1°, 27.6 ± 0.1°, and 29.8 ± 0.1°.

[0027] In some embodiments, form A has an XRPD diagram that is substantially as shown in FIG1A or FIG1E.

[0028] In some embodiments, form A is characterized by having two endothermic peaks at about 150°C and about 178°C according to differential scanning calorimetry (DSC).

[0029] In some embodiments, form A has a DSC thermogram that is substantially as shown in FIG1B.

[0030] In some embodiments, form A is characterized by a triclinic crystal system and a space group of P1, having the following cell parameters: (a) about 13.644 Å, (b) about 14.070 Å, (c) about 15.012 Å, (α) about 112.0202(3)°, (β) about 104.6821(3)°, and (γ) about 93.6507(2)°.

[0031] In a second aspect, this document discloses the crystalline form of compound 1, which is designated as an anhydrous form B.

[0032] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has a diffraction peak at an angle of 14.4 ± 0.1°.

[0033] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1° and 17.5 ± 0.1°.

[0034] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1°, 17.5 ± 0.1° and 18.4 ± 0.1°.

[0035] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

[0036] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.2 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

[0037] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.7 ± 0.1°, 7.2 ± 0.1°, 13.8 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

[0038] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.7 ± 0.1°, 7.2 ± 0.1°, 13.8 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

[0039] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.7 ± 0.1°, 7.2 ± 0.1°, 11.6 ± 0.1°, 12.2 ± 0.1°, 13.3 ± 0.1°, 13.8 ± 0.1°, 14.4 ± 0.1°, 15.7 ± 0.1°, 16.2 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1°, 19.6 ± 0.1°, 19.9 ± 0.1°, 23.0 ± 0.1°, and 24.9 ± 0.1°.

[0040] In some embodiments, an XRPD diagram is provided, which is substantially as shown in FIG2A or FIG2D.

[0041] In some embodiments, form B is characterized by having an endothermic peak at approximately 187°C according to differential scanning calorimetry (DSC).

[0042] In some embodiments, form B has a DSC thermogram that is substantially as shown in FIG2B.

[0043] In a third aspect, this document discloses the crystalline form of compound 1, which is designated as the anhydrous form U.

[0044] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1° and 24.3 ± 0.1°.

[0045] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 15.6 ± 0.1° and 24.3 ± 0.1°.

[0046] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 15.6 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

[0047] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

[0048] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

[0049] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

[0050] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

[0051] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

[0052] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

[0053] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 17.5 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

[0054] In some embodiments, the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 16.1 ± 0.1°, 17.0 ± 0.1°, 17.5 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, 21.6 ± 0.1°, and 24.3 ± 0.1°.

[0055] In some embodiments, the X-ray powder diffraction pattern of this crystalline form has angular values ​​of 7.0 ± 0.1°, 9.4 ± 0.1°, 10.2 ± 0.1°, 10.7 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 13.9 ± 0.1°, 14.9 ± 0.1°, 15.0 ± 0.1°, 15.6 ± 0.1°, 16.1 ± 0.1°, 17.0 ± 0.1°, 17.1 ± 0.1°, 17.5 ± 0.1°, 18.0 ± 0.1°, 18.4 ± 0.1°, 18.9 ± 0.1°, 19.2 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, and 20.5 ± 0.1°. Diffraction peaks are present at ± 0.1°, 21.2 ± 0.1°, 21.6 ± 0.1°, 22.3 ± 0.1°, 22.6 ± 0.1°, 22.9 ± 0.1°, 23.6 ± 0.1°, 24.3 ± 0.1°, 25.7 ± 0.1°, 25.8 ± 0.1°, 26.1 ± 0.1°, 27.6 ± 0.1°, 28.5 ± 0.1°, 28.9 ± 0.1°, and 29.3 ± 0.1°.

[0056] In some embodiments, form U has an XRPD diagram that is substantially as shown in FIG21A.

[0057] In some embodiments, form U is characterized by having an endothermic peak at approximately 164°C according to differential scanning calorimetry (DSC).

[0058] In some embodiments, form U has a DSC thermogram that is substantially as shown in FIG21B.

[0059] In the fourth aspect, the crystalline form of compound 1 is specified as form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, form Q, form R, form S or form T.

[0060] In some embodiments, forms C, D, E, F, G, H, I, J, K, L, M, N, O, P, Q, R, S and T each have an XRPD diagram substantially as shown in Figures 3A, 4A, 5A, 6A, 7A, 8A, 9A, 10A, 11A, 12A, 13A, 14A, 15A, 16A, 17, 18A, 19A and 20A.

[0061] In some embodiments of all the above aspects, the crystalline form is at least 40%, 50%, 60%, 70%, 80%, 90% or 95% crystalline.

[0062] In the fifth aspect, this article discloses the amorphous form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide (compound 1).

[0063] In some embodiments, the amorphous form of compound 1 has an XRPD diagram that is substantially as shown in FIG22A.

[0064] In some embodiments, the amorphous form of compound 1 is characterized by having a glass transition signal (moderate) at about 127°C.

[0065] In some embodiments, the amorphous form of compound 1 contains no more than 1%, 2%, 3%, 4%, 5% or 10% of the crystalline form of compound 1.

[0066] In a sixth aspect, this document discloses a pharmaceutical composition comprising (a) a therapeutically effective amount of compound 1 in solid form, preferably in crystalline form or in amorphous form of compound 1 disclosed herein; and (b) one or more pharmaceutically acceptable excipients.

[0067] In some embodiments, the crystalline form of compound 1 is the crystalline form of the EtOAc solvate of compound 1, containing about 1 mol of EtOAc per mol; and is the anhydrous form of compound 1.

[0068] In some embodiments, the crystalline form of compound 1 is form A, form B or form U of compound 1.

[0069] In some embodiments, the crystalline form of compound 1 is form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, form Q, form R, form S or form T.

[0070] In a seventh aspect, this document discloses a method for preparing a pharmaceutical solution of compound 1, the method comprising dissolving compound 1 in a solid form, preferably in a crystalline form as described in claim 1, or in an amorphous form, in a pharmaceutically acceptable solvent or a mixture of solvents.

[0071] In an eighth aspect, this article discloses a method for treating diseases associated with Bcl-2 protein inhibition, the method comprising administering to a subject a therapeutically effective amount of the crystalline form, amorphous form, or pharmaceutical composition of the compound 1 disclosed herein.

[0072] In some embodiments, the disease associated with Bcl-2 protein inhibition is an dysregulated apoptotic disease. In some preferred embodiments, the disease associated with Bcl-2 protein inhibition is a neoplastic, prothrombotic, immune, or autoimmune disease.

[0073] In some embodiments, the crystalline form of compound 1 is form A, form B or form U of compound 1.

[0074] In some embodiments, the crystalline form of compound 1 is the crystalline form of the EtOAc solvate of compound 1, containing about 1 mol of EtOAc per mol; or it is the anhydrous form of compound 1.

[0075] In some embodiments, the crystalline form of compound 1 is form C, form D, form E, form F, form G, form H, form I, form J, form K, form L, form M, form N, form O, form P, form Q, form R, form S or form T.

[0076] In some embodiments, the therapeutically effective dose is administered orally at a dose of about 1 mg to about 640 mg of compound once a day.

[0077] In some embodiments, the subject is a human being.

[0078] In some embodiments, form A is obtained by a method comprising any of the following procedures: a) dissolving compound 1 in DCM, removing DCM, and adding EA to obtain form A; b) dissolving compound 1 in DCM, concentrating, adding EA, and exchanging DCM with EA, MeOH, and EA respectively to obtain form A; c) dissolving compound 1 in EA, heating, and cooling to obtain form A; or d) dissolving compound 1 in a THF / EtOAc (1:2, v / v) solvent mixture and evaporating to obtain form A.

[0079] In some embodiments, form B is obtained by a method comprising any of the following procedures: a) dissolving compound 1 in acetone and evaporating the solvent to obtain the desired crystalline form; b) heating forms A, C, and O to about 160°C and cooling back to RT to obtain form B; c) gradually isothermally heating form A to about 100°C to obtain form B; d) heating form D or J to about 130°C and isothermally to obtain form B; or e) adding form K to heptane, refluxing at about 100°C and cooling to obtain form B.

[0080] In some embodiments, form U is obtained by a method comprising any of the following procedures: a) dissolving compound 1 in DCM, adding n-heptane in batches and stirring to obtain form U; b) dissolving compound 1 in a mixture of DCM / n-heptane (1:1, v / v) and stirring to obtain form U.

[0081] In some embodiments, form A and / or form B are obtained by a method including adding seed crystals to a solution system.

[0082] In some embodiments, the amorphous form is obtained by a method comprising any of the following procedures: a) dissolving compound 1 in DCM and drying to obtain the amorphous form; or b) dissolving compound 1 in a mixture of solvents containing DCM and drying to obtain the amorphous form.

[0083] In some embodiments, the amorphous form is obtained by a method including dissolving compound 1 in a solid form, preferably in the crystalline form of compound 1.

Implementation Method

[0085] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All patents, patent applications, and publications mentioned herein are incorporated herein by reference.

[0086] As used herein, the term "solvent" refers to the crystalline form of compound 1 containing a solvent.

[0087] As used herein, the interchangeable terms "subject," "individual," or "patient" refer to any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, primates, and humans. In some embodiments, the patient is a human. In some embodiments, the subject has experienced and / or exhibited at least one symptom of a disease or disorder to be treated and / or prevented. In some embodiments, the subject is suspected of having polytyrosine kinase-related cancer.

[0088] As used herein, a "therapeuticly effective amount" of the crystalline form of the salt of compound 1 is an amount sufficient to improve or alleviate symptoms in some way, or to stop or reverse the progression of the disease, or to negatively regulate or inhibit the activity of polytyrosine kinase. Such an amount may be administered as a single dose or according to a regimen, thereby being effective.

[0089] As used herein, the term "form" is used to describe a crystalline form and may be used interchangeably with the term "type". The term "crystalline form" refers to a crystalline solid form. In some embodiments, the crystalline form of a substance may be substantially free of amorphous and / or other crystalline forms. In some embodiments, the crystalline form of a substance may contain less than about 1%, less than about 2%, less than about 3%, less than about 4%, less than about 5%, less than about 6%, less than about 7%, less than about 8%, less than about 9%, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, or less than about 50% (by weight) of one or more amorphous and / or other crystalline forms. In some embodiments, the crystalline form of a substance may be physically and / or chemically pure. In some embodiments, the crystalline form of the substance may be about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, or about 90% physically and / or chemically pure.

[0090] As used herein, "amorphous form" refers to particles without a defined structure, such as those lacking a crystalline structure. Unless otherwise stated, the term "amorphous" or "amorphous form" means that the substance, component, or product in question is determined by X-ray diffraction to be substantially non-crystalline. In particular, the term "amorphous form" describes a disordered solid form, i.e., a solid form lacking long-range crystalline order. In some embodiments, the amorphous form of a substance may be substantially free of other amorphous and / or crystalline forms. In some embodiments, the amorphous form of a substance may contain less than about 1%, less than about 2%, less than about 3%, less than about 4%, less than about 5%, less than about 10%, less than about 15%, less than about 20%, less than about 25%, less than about 30%, less than about 35%, less than about 40%, less than about 45%, or less than about 50% (by weight) one or more other amorphous and / or crystalline forms. In some embodiments, the amorphous form of a substance may be physically and / or chemically pure. In some embodiments, the amorphous form of the substance is about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, or about 90% physically and / or chemically pure.

[0091] As used herein, “treatment” means any means of improving or otherwise beneficially altering the symptoms or pathology of a condition, disorder, or disease. Treatment also includes any pharmaceutical use of the components of this article.

[0092] As used herein, improvement of symptoms of a particular condition by administration of a particular pharmaceutical composition means any relief attributable to or related to administration of the composition, whether permanent or temporary, lasting or transient.

[0093] As used herein, the term "about" in the context of XRPD peak position refers to the inherent variability of the peak, which depends on the instrument calibration, the method used to prepare the crystalline form of the present invention, the age of the crystalline form, and the type of instrument used in the analysis. The variability of the instrument used for XRPD analysis is about ± 0.1 °2θ.

[0094] As used herein, the term "about" when referring to the onset of an endothermic peak in DSC refers to the inherent variability of the peak, which depends on the instrument calibration, the method used to prepare the sample of the present invention, and the type of instrument used in the analysis. The variability of the instrument used for DSC analysis is about ±1°C. General Methods

[0095] Unless otherwise stated, the general approach outlined below is used in the exemplary instances.

[0096] I. Crystallization Technology

[0097] The crystalline forms disclosed herein can be prepared using a variety of methods well known to those skilled in the art, including crystallization or recrystallization from a suitable solvent or by sublimation. A variety of techniques, including those in the exemplary examples, can be used for crystallization or recrystallization, including evaporating a solvent or solvent mixture miscible or immiscible with water, seeding crystals in a supersaturated solution, lowering the temperature of the solvent mixture, or freeze-drying the solvent mixture.

[0098] The crystallization disclosed herein can be performed with or without a seed crystal. The seed crystal may be derived from any prior batch of the desired crystallization forms: for example, forms C, D, E, F, G, H, I, J, K, L, M, N, O, P, Q, R, S, or T. Abbreviations and Acronyms; Instruments and Parameters

[0099] For XRPD analysis, unless otherwise specified, a PANalytical Empyrean and X'Pert3 X-ray powder diffractometer were used to characterize the physical forms obtained in this disclosure. The XRPD parameters used are listed below. parameter XRPD Model Empyrean X'Pert 3 X-ray wavelength Cu, kα, Kα1 (Å): 1.540598, Kα2 (Å): 1.544426 Kα2 / Kα1 intensity ratio: 0.50 Cu, kα, Kα1 (Å): 1.540598, Kα2 (Å): 1.544426 Kα2 / Kα1 intensity ratio: 0.50 X-ray tube setup 45 kV, 40 mA 45 kV, 40 mA Diverging slit Automatic 1 / 8° Scanning mode Continuous Continuous Scan range (°2θ) 3-40 3-40 Step scan time (s) 17.8 46.7 Step size (°2TH) 0.0167 0.0263 Test time (s) 5 min 30 s About 5 minutes (5 minutes 04 seconds)

[0100] For XRPD analysis, a Bruker D8 advanced X-ray powder diffractometer or equivalent was also used to characterize forms A and U. The XRPD parameters used are listed below. Tube Cu, kα, Kα (Å): 1.540598, dynamo Voltage: 40 kV; Current: 40 mA Fixed incident beam optics Main cable slit: 2.5° Secondary Solar slit: 2.5° Diverging slit: 0.60 mm Slit: Fixed detector Detector name: Lynxeye (1D mode) PSD OPENING 2.1o Scanning mode Continuous PSD Fast Scan range (°2θ) 4-40° Scan type Coupled Two Theta / Theta Increment 0.02° Time / Step 0.12 s / step

[0101] Unless otherwise specified, TGA and DSC were used to characterize the physical forms obtained in this disclosure, wherein TGA data were collected using a TA Q500 / Q5000 TGA from TA Instruments, and DSC was performed using a TA Q200 / Q2000 DSC from TA Instruments. The detailed parameters used are listed below. parameter TGA DSC mDSC method Ramp Ramp adjust Sample tray Aluminum, open type Aluminum, press-fit Aluminum, press-fit temperature RT - Required temperature 25°C - Required temperature 16°C - Required temperature heating rate 10°C / min 10°C / min 3°C / min purge gas N2 N2 N2

[0102] For TGA and DGA analysis of forms A or U, several instruments were used for the test, including a NETZSCH TG 209 F1 instrument for collecting TGA data and a TA Q 20 or TA DSC 250 instrument for DSC. The detailed parameters used are listed below. parameter TGA DSC method Ramp Ramp Sample tray Aluminum, open type Aluminum, sealing temperature RT - Required temperature RT - Required temperature heating rate 10°C / min 10°C / min purge gas N2 N2

[0103] The DVS obtained in this disclosure was measured by SMS (Surface Measurement Systems) DVS Intrinsic, unless otherwise specified (Method A). The relative humidity at 25°C was calibrated according to the deliquescence points of LiCl, Mg(NO3)2, and KCl. The parameters for DVS detection are listed below. parameter DVS temperature 25°C Sample size 10 - 20 mg Gas and flow rate N2, 200 mL / min dm / dt 0.002% / min Shortest dm / dt stability duration 10 min Longest balancing time 180 min RH range 70%RH-95%RH-0%RH-95%RH RH step size 10% (0%RH-90%RH, 90%RH-0%RH) 5% (90%RH-95%RH, 95%RH-90%RH)

[0104] The DVS of forms A and U are also measured using SMS (Surface Measurement System) DVS Intrinsic (Method B). The relative humidity at 25°C is calibrated according to the deliquescence points of LiCl, Mg(NO3)2, and KCl. The parameters for DVS detection are listed below. parameter DVS temperature 25°C Gas and flow rate N2, 200 mL / min dm / dt < 0.01 min Shortest dm / dt stability duration 60 min Longest balancing time 180 min RH range Cycle: 40%-0%-95%-0%-40%RH

[0105] Single-crystal X-ray diffraction data were collected at 120 K using a Rigaku XtaLAB Synergy R (CuK radiation, 1.54184 Å) diffractometer. The instrument parameters are listed below.

[0106] The following examples are intended to further illustrate certain embodiments of the present invention, and are not intended to limit the scope of the present invention. Examples

[0107] Methods for producing the Bcl-2 inhibitor 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-(((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide (Compound 1) are known. For example, International Publication No. WO 2019 / 210828 provides a detailed synthetic route for the preparation of Compound 1. Example 1A: Preparation of Compound 1 in Form A (Form A)

[0108] Compound 1 (40 g) was dissolved in DCM (120 mL). The solution was concentrated to dryness, and EA (250 mL) was added. The resulting mixture was heated to 60°C-70°C, slowly cooled to 15°C-25°C, and then filtered. The resulting filter cake was dried at 40°C-50°C for 16 hours to obtain compound 1 in form A (about 40 g), which can be used as seed crystals.

[0109] Compound 1 (8.1 kg) was dissolved in DCM (58 kg) at 20°C-30°C. After concentrating the solution to half the volume of the mixture, EA (45 kg) was added to the solution, along with seed crystals (0.035 kg). After stirring at 20°C-30°C for 1 hour, the solution was concentrated to exchange the EA solvent mixture with EA three times (43 kg + 43 kg + 24 kg). The mixture was heated to 60°C-70°C and stirred for 2 hours, then slowly cooled to 15°C-25°C.

[0110] MeOH (32 kg) was introduced into the resulting mixture at 45°C-55°C and stirred for 16 hours. After three solvent exchanges with MeOH (20 kg + 21 kg + 20 kg) and three solvent exchanges with EA (23 kg + 47 kg + 40 kg), the mixture was returned to the EA solution. The mixture was heated to 60°C-70°C and stirred for 2.5 hours, then slowly cooled to 15°C-25°C. The resulting mixture was slowly cooled to 15°C-25°C and filtered. The resulting filter cake was washed with EA (9 kg) and dried at 45°C-55°C for 18.5 hours to give a product as a yellow solid. After sieving the solid, a total of 7.36 kg of compound form A was obtained.

[0111] X-ray powder diffraction (XRPD) pattern (performed on a Bruker D8 advanced X-ray powder diffractometer) was used to characterize form A, indicating that form A is crystalline, see Figure 1A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 1A. [Table 1A]. XRPD pattern of compound 1, form A Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.928 12.74962 99.9 7.431 11.88697 5.9 8.762 10.08430 9.9 10.603 8.33666 34.4 10.919 8.09612 15.1 12.359 7.15614 5.4 12.654 6.98993 82.8 13.090 6.75786 4.8 13.363 6.62063 39.3 13.760 6.43061 37.7 14.126 6.26474 69.4 14.701 6.02105 13.5 14.936 5.92683 43.9 15.350 5.76762 4.6 16.197 5.46788 15.9 16.456 5.38232 84.2 16.928 5.23335 87.0 17.455 5.07668 10.2 18.179 4.87613 5.9 18.456 4.80356 46.6 19.142 4.63277 100.0 19.524 4.54294 90.8 20.693 4.28895 35.7 20.737 4.27996 45.1 21.144 4.19839 12.4 21.796 4.07435 24.1 22.380 3.96928 48.8 22.837 3.89098 10.2 23.251 3.82265 10.6 23.785 3.73795 49.8 24.123 3.68627 6.2 24.497 3.63087 26.4 25.791 3.45163 24.5 26.719 3.33379 4.7 27.108 3.28677 10.0 27.592 3.23023 10.0 29.751 3.00059 9.1

[0112] The 1H NMR spectrum of compound form A is shown in Figure 1D. The DSC / TGA curves (performed on a NETZSCH TG 209 F1 instrument and TA Q 20) show an observed weight loss of 8.8% up to 160°C, and two endothermic peaks at 149.6°C and 178.2°C (Figures 1B and 1C). The XRPD overlay plot shows that after heating to 160°C, form A transforms into form B, which is then cooled back to RT and re-exposed to air. Combining the TGA data and 1H NMR results, form A is presumably an EtOAc solvent.

[0113] In a nitrogen atmosphere, compound 1, form A, was obtained by gradual isothermal heating using TGA. When the weight loss reached 0.02%, the system equilibrated at a certain temperature until the weight loss was < 0.002%. The results showed that the TGA weight loss matched the weight loss detected by linear heating after gradually heating form A to 100°C. After cooling back to RT, form B with low crystallinity was obtained.

[0114] The DVS cycle was performed at 25°C (Method B). The adsorption and desorption systems were correctable throughout the DVS cycle. The water absorption rate was 0.4% at 95% RH humidity. Compound 1, form A, is slightly hygroscopic. Example 1B: Preparation of compound 1, form A

[0115] Compound 1 (7.0 g) was added to EA (140 mL), and then heated to reflux for 2 hours. The mixture was slowly cooled to room temperature (RT) and stirred for 0.5 hours, filtered, washed with EA, and dried under reduced pressure to give the product (4.9 g). Example 1C: Preparation of Compound 1 in Single Crystal Form A

[0116] Compound 1 (2.8 mg) was dissolved in 0.5 mL of a THF / EtOAc (1:2, v / v) solvent mixture. After slow evaporation, single crystals of compound 1 in form A were obtained.

[0117] The single crystal of compound 1, form A (EtOAc solvate), was characterized by SCXRD. The calculated XRPD of the single crystal structure was almost identical to the experimental XRPD of the form A single crystal (Fig. 1E).

[0118] The single crystal was analyzed using a single-crystal X-ray diffractometer. The crystal system of the single crystal is triclinic, and the space group is P1. The cell parameters are: {a = 13.64421(4) Å, b = 14.07005(4) Å, c = 15.01208(4) Å, α = 112.0202(3)°, β = 104.6821(3)°, γ = 93.6507(2)°, V = 2543.673(14) Å3}.

[0119] The asymmetric unit of the single-crystal structure consists of two compound 1 molecules and two EtOAc molecules, indicating that the crystal is an EtOAc solvate and the molar ratio of compound 1 to EtOAc is 1:1. Furthermore, adjacent compound 1 molecules are connected to each other via intermolecular hydrogen bonds. Example 2A: Preparation of compound 1 in form B

[0120] Compound 1 (20 mg) was dissolved in acetone. The mixture was filtered, and the resulting clear solution was then slowly evaporated at RT to obtain form B.

[0121] The XRPD plot was used to characterize form B, indicating that form B is crystalline, see Figure 2A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 2A. [Table 2A]. XRPD plot of compound 1, form B. Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.65 13:30 21.12 7.22 12.24 100.00 11.58 7.64 7.80 12.17 7.27 7.39 13.28 6.67 9.80 13.77 6.43 11.34 14.42 6.14 32.09 15.67 5.66 8.98 16.22 5.47 8.91 17.54 5.06 54.27 18.36 4.83 23.45 19.60 4.53 10.71 19.92 4.46 9.02 23.03 3.86 3.35 24.87 3.58 8.92

[0122] The TGA / DSC curves showed a weight loss of 3.3% at up to 110°C, and two endothermic peaks at 107.7°C and 187.3°C (peaks) before decomposition (Figure 2B). Approximately 2.2% acetone was observed in the 1H NMR spectrum (Figure 2C). No change in form B was observed after heating to 160°C.

[0123] VT-XRPD results showed that no change in form was observed after heating form B to 150°C and cooling it back to 30°C in a N2 atmosphere, indicating that form B is an anhydrous compound. It is speculated that the acetone detected in 1H NMR was due to solvent residue.

[0124] Furthermore, form B with high crystallinity can be obtained by heating it to 160°C, cooling it back to RT, and then reheating it to 160°C, as shown in Figure 2D. TGA / DSC curves show a weight loss of 2.8% observed up to 150°C, and an endothermic peak at 186.5°C (peak) before decomposition. No acetone signal was detected in the 1H NMR spectrum. Example 2B: Preparation of Form B of Compound 1

[0125] Form B of compound 1 was obtained by any of the following steps: 1) Heating form A to 160°C and then naturally cooling to RT; 2) Gradually isothermally heating form A to 100°C; 3) Heating form D to 130°C and holding isothermally for 30 min; or 4) Heating form J to 130°C and holding isothermally for 30 min; 5) Heating form B to 160°C and cooling back to RT, then heating to 160°C. Example 2C: Preparation of form B of compound 1

[0126] Compound 1, form K (6.0 g), was added to heptane (100 mL) and refluxed at approximately 100°C for 24 h to induce slurry formation. The mixture was cooled to RT and filtered, washed with heptane, and dried under reduced pressure to give the product (5.5 g). Example 3A: Preparation of compound 1, form C (form C)

[0127] Compound 1 (20 mg) was dissolved in MEK. The mixture was filtered, and the resulting clear solution was then slowly evaporated at RT to obtain form C.

[0128] The X-ray powder diffraction (XRPD) pattern was used to characterize form C, indicating that form C is in a crystalline form, see Figure 3A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 3A. [Table 3A]. XRPD pattern of form C of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.91 12.79 100.00 10.61 8.34 53.40 12.65 7.00 36.44 13.88 6.38 55.60 14.23 6.22 38.85 14.99 5.91 12.75 16.43 5.39 99.90 16.75 5.29 39.88 18.42 4.82 9.40 18.81 4.72 8.65 19.17 4.63 15.78 19.53 4.55 33.14 20.97 4.24 16.62 21.50 4.13 13.42 21.97 4.05 16.02 23.42 3.80 8.88 23.97 3.71 14.93 24.72 3.60 19.55 27.49 3.25 7.94

[0129] The TGA / DSC curves showed a weight loss of 8.1% at up to 160°C, and two endothermic peaks at 142.5°C and 177.3°C (Figure 3B). The 1H NMR spectrum (Figure 3C) showed a theoretical weight loss of 5.4% for MEK, which is lower than the TGA weight loss and is presumed to be due to solvent loss during storage before the 1H NMR test. To clarify whether the weight loss was due to solvent absorption, a heating experiment was performed on form C.

[0130] XRPD comparison showed that after heating to 160°C, cooling back to RT, and re-exposure to air, form C transformed into form B, which has weak crystallinity. It is speculated that form C is a MEK solvate. Example 4A: Preparation of form D of compound 1 (form D)

[0131] Amorphous compound 1 (20 mg) was suspended in IPAc. The suspension was slurried by stirring at RT for 1–7 days to obtain form D.

[0132] The X-ray powder diffraction (XRPD) pattern was used to characterize the obtained form D, indicating that form D is a crystalline form, see Figure 4A. The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 4A. [Table 4A]. XRPD pattern of form D of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.50 13.59 100.00 7.25 12.20 6.55 8.86 9.99 14.43 9.80 9.02 0.98 10.74 8.24 3.12 12.98 6.82 27.15 13.66 6.48 15.70 14.23 6.23 14.18 14.63 6.06 19.43 15.68 5.65 3.95 16.39 5.41 2.92 17.25 5.14 12.56 17.82 4.98 43.14 18.21 4.87 23.91 18.90 4.70 6.63 20.09 4.42 19.12 20.46 4.34 14.52 21.51 4.13 5.51 22.22 4.00 4.07 23.16 3.84 2.14 23.83 3.73 4.03 24.32 3.66 3.56 24.80 3.59 4.84 26.00 3.43 4.53 26.84 3.32 3.09 28.99 3.08 3.23

[0133] TGA / DSC data showed a weight loss of 7.2% at up to 130°C, and three endothermic peaks at 108.4°C, 160.1°C, and 177.3°C (Figure 4B). ¹H NMR spectroscopy showed a theoretical IPAc content of 5.4%, indicating that some solvent loss may have occurred during storage (Figure 4C).

[0134] In the XRPD overlay plot of the heating experiment, the form change of form D was observed after heating to 165°C and cooling back to RT. Therefore, it is speculated that form D is an IPAc solvate.

[0135] Furthermore, heating experiments showed that heating form D to 130°C and holding it isothermally for 30 min yielded form B with lower crystallinity and an additional peak. Example 5A: Preparation of form E of compound 1 (form E)

[0136] Compound 1 (20 mg) was dissolved in anisole. The mixture was filtered, and the resulting clear solution was then slowly evaporated at RT to obtain form E.

[0137] The X-ray powder diffraction (XRPD) pattern was used to characterize the obtained form E, indicating that form E is crystalline, see Figure 5A. The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 5A. [Table 5A]. XRPD pattern of form E of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.96 12.70 100.00 8.60 10.29 1.95 10.90 8.12 11.86 12.48 7.09 6.29 13.16 6.73 2.24 13.91 6.37 38.41 14.79 5.99 6.37 15.18 5.84 2.37 16.09 5.51 2.66 16.91 5.24 10.97 17.16 5.17 11.50 18.09 4.90 4.50 18.54 4.78 2.90 19.14 4.64 11.69 19.51 4.55 1.34 20.13 4.41 2.79 21.08 4.21 10.74 21.73 4.09 3.29 22.09 4.02 6.11 22.62 3.93 2.01 22.96 3.87 2.07 24.01 3.71 2.14 25.10 3.55 4.72 25.81 3.45 1.59 27.18 3.28 3.39 27.75 3.21 2.10 28.17 3.17 1.39 29.67 3.01 1.15

[0138] The TGA curve showed a weight loss of 11.9% at temperatures up to 180°C, and the DSC curve showed an endothermic peak at 157.4°C (peak) before decomposition (Figure 5B). Based on the 1H NMR spectrum (Figure 5C), approximately 17.1% anisole was determined, which is higher than the TGA weight loss and is presumably due to solvent inhomogeneity. Heating experiments showed a decrease in crystallinity after heating form E to 170°C and then cooling it back, indicating that the endothermic peak on the DSC curve may be a melting signal. Form E is presumably an anisole solvate. Example 6A: Preparation of Form F of Compound 1

[0139] Amorphous compound 1 (20 mg) was suspended in 0.5 mL EtOH and stirred at 50°C to obtain F.

[0140] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form F, indicating that form F is in a crystalline form, see Figure 6A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 6A. [Table 6A]. XRPD pattern of form F of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.84 12.92 100.00 8.66 10.21 3.90 10.50 8.43 17.22 10.85 8.16 4.13 12.55 7.05 40.33 13.26 6.68 6.78 13.66 6.48 40.53 14.02 6.31 39.46 14.56 6.08 7.50 14.81 5.98 11.14 16.34 5.43 28.89 16.83 5.27 21.43 17.33 5.12 3.01 18.35 4.84 10.27 19.05 4.66 18.00 19.41 4.57 30.18 20.35 4.36 7.25 20.68 4.30 23.86 21.07 4.22 2.05 21.68 4.10 3.97 22.26 3.99 8.26 22.73 3.91 4.02 23.10 3.85 3.15 23.73 3.75 14.37 24.39 3.65 15.77 24.89 3.58 1.93 25.69 3.47 5.86 26.55 3.36 2.66 26.99 3.30 7.78 27.48 3.25 2.08 28.22 3.16 1.76 29.64 3.01 2.68

[0141] TGA / DSC curves show a weight loss of 0.8% at up to 80°C, a broad peak near 69.7°C, and two endothermic peaks at 156.8°C and 177.8°C (peaks) before decomposition (Figure 6B). No EtOH signal was detected in the 1H NMR spectrum (Figure 6C). Heating experiments showed no change in form when form F was heated to 80°C, and diffraction peaks of form B were detected after heating form F to 150°C and 165°C. According to VT-XRPD results, no change in form was observed after heating form F to 100°C in N2 and cooling it back to 30°C, indicating that form F is anhydrous. It is speculated that the broad endothermic reaction observed in DSC at 69.7°C is due to the loss of residual solvent or moisture, while the endothermic reaction at 156.8°C may be related to form transformation at higher temperatures. Example 7A: Preparation of form G of compound 1 (form G)

[0142] Amorphous compound 1 (20 mg) was suspended in MTBE. The suspension was slurried by stirring at RT for 1–7 days to obtain form G.

[0143] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form G, indicating that form G is in a crystalline form, see Figure 7A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 7A. [Table 7A]. XRPD pattern of form G of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 5.98 14.78 35.87 7.17 12.33 69.66 8.71 10.15 24.51 10.00 8.84 36.98 12.76 6.94 58.14 13.16 6.73 57.59 13.61 6.51 100.00 14.18 6.25 36.45 15.66 5.66 31.51 16.02 5.53 41.86 16.62 5.33 29.84 18.10 4.90 33.05 18.62 4.76 67.01 19.08 4.65 53.52 19.43 4.57 58.83 19.81 4.48 50.65 20.38 4.36 27.44 20.80 4.27 33.06 21.81 4.08 34.69 22.23 4.00 29.86 23.62 3.77 46.42 25.82 3.45 14.89 26.52 3.36 8.36 27.80 3.21 8.42 28.83 3.10 6.55 30.41 2.94 7.11

[0144] TGA / DSC results showed a 4.6% weight loss at up to 160°C, a weak endothermic peak at 117.2°C before decomposition, and a strong endothermic peak at 157.7°C (Figure 7B). Based on the 1H NMR results (Figure 7C), the theoretical weight of MTBE was calculated to be 5.1%. XRPD coverage plots before and after heating showed a significant decrease in crystallinity after heating. Form G is presumed to be an MTBE solvent. Example 8A: Preparation of Form H of Compound 1 (Form H)

[0145] Amorphous compound 1 (20 mg) was suspended in ACN. The suspension was slurried at RT for 1-7 days to obtain form H.

[0146] X-ray powder diffraction (XRPD) patterns were used to characterize form H, indicating that form H is crystalline, see Figure 8A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 8A. [Table 8A]. XRPD pattern of form H of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.29 14.05 14.03 6.91 12.79 37.76 7.28 12.14 86.45 7.64 11.57 38.25 8.87 9.96 36.86 9.05 9.78 26.74 9.63 9.18 2.53 10.78 8.21 18.31 10.94 8.09 17.12 12.54 7.06 23.74 13.10 6.76 67.22 13.83 6.40 21.51 14.68 6.03 55.92 14.99 5.91 100.00 15.39 5.76 22.45 16.54 5.36 11.39 17.76 4.99 16.90 18.16 4.89 22.35 18.74 4.74 23.28 19.46 4.56 40.81 20.02 4.44 41.97 20.88 4.26 11.42 22.09 4.02 15.85 23.52 3.78 20.04 25.33 3.52 4.14 28.98 3.08 2.57

[0147] TGA / DSC curves showed a 1.2% weight loss at up to 170°C, and three endothermic peaks at 60.1°C, 162.9°C, and 179.5°C (Figure 8B) before decomposition. No ACN signal was detected in the 1H NMR results (Figure 8C), indicating that form H may be an anhydrous / hydrated form. Example 9A: Preparation of form I of compound 1 (form I)

[0148] Amorphous compound 1 (20 mg) was suspended in 0.5 mL IPA and stirred at 50°C to obtain form I.

[0149] X-ray powder diffraction (XRPD) pattern is used to characterize the obtained form I, which shows that form I is crystalline, see Figure 9A. Position [°2θ] Interplanar spacing [Å] Relative strength [%] 3.58 24.68 4.38 7.09 12.47 100.00 8.03 11.01 2.57 9.61 9.20 1.69 10.67 8.29 0.94 12.21 7.25 7.50 12.49 7.09 8.01 14.02 6.32 10.43 14.68 6.04 1.56 15.44 5.74 6.62 16.32 5.43 1.69 16.95 5.23 4.71 17.69 5.01 2.53 18.89 4.70 7.02 19.90 4.46 3.08 20.56 4.32 14.49 21.44 4.14 2.06 22.07 4.03 2.72 22.97 3.87 1.97 24.06 3.70 1.61

[0150] TGA / DSC curves showed a weight loss of 2.1% at up to 120°C, and two endothermic peaks at 134.0°C and 159.7°C before decomposition (Fig. 9B). An IPA peak was observed in the 1H NMR spectrum (Fig. 9C), with a calculated content of 3.2%. XRPD comparison showed a significant decrease in crystallinity of form I during heating experiments. Form I is presumed to be an IPA solvate. Example 9B: Preparation of Compound 1 Form I. Form I obtained by slow evaporation in acetone showed the same XRPD curve as form I in Example 9A. TGA weight losses were observed in both steps (1.9% at up to 110°C and 2.7% from 110°C to 200°C, see Fig. 9D), and two endothermic peaks were observed in the DSC thermogram before decomposition at 78.0°C and 160.3°C. 1H NMR (Figure 9E) results showed that the acetone content in the sample was 2.8%.

[0151] Based on the heating experiment results, no change in form I from acetone was observed after heating to 130°C, but an amorphous sample was observed when the heating temperature reached 180°C. Combining TGA and 1H NMR data, the first step of the TGA weight loss may be the desorption of volatile components, while the second step of the weight loss may be due to the loss of acetone, leading to a transition to the amorphous phase. Therefore, it is speculated that form I from acetone is an acetone solvate.

[0152] Since different solvates have the same XRPD plot as form I, it is speculated that isomorphism occurred during the formation of form I. Example 10A: Preparation of compound 1, form J (form J)

[0153] Amorphous compound 1 (20 mg) was suspended in 2-MeTHF / n-heptane (1:1, v / v). The suspension was slurried by stirring at RT for 1–7 days to obtain form J.

[0154] The X-ray powder diffraction (XRPD) pattern was used to characterize form J, indicating that form J is a crystalline form, see Figure 10A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 10A. [Table 10A]. XRPD pattern of form J of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.98 12.67 100.00 8.66 10.21 10.03 10.73 8.24 51.79 12.53 7.06 61.11 13.10 6.76 16.09 13.90 6.37 30.35 14.18 6.25 27.31 14.96 5.92 36.10 15.83 5.60 8.80 16.57 5.35 64.29 16.82 5.27 55.14 17.34 5.11 5.84 18.44 4.81 31.33 19.23 4.62 67.51 20.28 4.38 19.13 20.54 4.32 14.82 21.03 4.23 33.70 21.59 4.12 22.00 21.97 4.05 6.20 22.42 3.97 16.54 23.24 3.83 11.51 23.89 3.72 19.15 24.85 3.58 21:30 25.89 3.44 6.76 27.39 3.26 5.43 28.69 3.11 2.02 29.67 3.01 3.21

[0155] TGA / DSC curves showed an 8.0% weight loss at up to 160°C, and two endothermic peaks at 125.3°C and 175.2°C (Figure 10B) before decomposition. ¹H NMR results (Figure 10C) showed 2-MeTHF and n-heptane signals in form J (theoretical weight loss: ~10.2%). XRPD coverage plots showed that form J transformed into form B after heating to 150°C and cooling back to RT. Based on TGA, ¹H NMR, and heating experimental data, form J is presumed to be a 2-MeTHF solvate.

[0156] Furthermore, form J was heated to 130°C, then held isothermally at 130°C for 30 min, and then cooled to RT. XRPD results showed that form B with low crystallinity was obtained. Example 11A: Preparation of form K of compound 1 (form K)

[0157] Amorphous compound 1 (20 mg) was suspended in methyl acetate. The suspension was stirred at RT for 1–7 days to slurry up to obtain form K.

[0158] The X-ray powder diffraction (XRPD) pattern was used to characterize the obtained form K, indicating that form K is in crystalline form, see Figure 11A. The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 11A. [Table 11A]. XRPD pattern of form K of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.90 12.81 100.00 8.69 10.17 13.04 10.56 8.38 36.54 12.63 7.01 68.65 13.85 6.39 38.66 14.21 6.23 51.09 14.88 5.95 22.63 16.33 5.43 45.89 16.82 5.27 47.13 18.42 4.82 27.88 18.99 4.67 36.24 19.54 4.54 47.49 20.80 4.27 21.69 21.78 4.08 11.61 22.29 3.99 19.17 23.73 3.75 21.88 24.50 3.63 19.06 25.85 3.45 8.16 27.53 3.24 7.32

[0159] TGA / DSC showed a weight loss of 5.8% at up to 120°C, and two endothermic peaks at 112.1°C and 177.7°C (peaks) before decomposition (Figure 11B). In 1H NMR (Figure 11C), a methyl acetate signal was observed, with a theoretical weight loss of ~2.5%.

[0160] The XRPD overlay plot of the heating test showed that form B, exhibiting weak crystallinity, was observed after heating form K to 120°C. As the XRPD overlay plot shows, form K transformed into form B, exhibiting low crystallinity, after approximately 5 weeks of RT storage in a sealed HPLC vial. It is speculated that form K is a methyl acetate solvate. Example 11B: Preparation of form K of compound 1

[0161] Compound 1 (8.0 g) was added to methyl acetate (100 mL), and then heated to 50°C for 2 hours. The mixture was cooled to RT and stirred for 16 hours. The mixture was filtered, washed with methyl acetate, and dried under reduced pressure to give the product (7.1 g). Example 12A: Preparation of Compound 1 in Form L (Form L)

[0162] Amorphous compound 1 (20 mg) was suspended in 0.5 mL of acetone / n-heptane (1:1, v / v) and stirred at 50°C to obtain form L.

[0163] X-ray powder diffraction (XRPD) patterns were used to characterize form L, indicating that form L is a crystalline form, see Figure 12A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 12A. [Table 12A]. XRPD pattern of form L of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.80 13.00 100.00 9.68 9.14 12.98 12.06 7.34 10.20 13.08 6.77 16.45 13.59 6.52 12.89 15.13 5.86 34.96 15.79 5.61 19.25 18.25 4.86 13.64 19.90 4.46 29.09 21.37 4.16 6.13 23.83 3.73 5.23

[0164] TGA / DSC curves show that a weight loss of 2.2% was observed at up to 100°C in the TGA curve; and in the DSC curve, multiple signals were detected, including four endothermic peaks at 53.7°C, 62.7°C, 76.3°C, and 162.1°C (peaks) before decomposition, and one exothermic peak at 89.6°C (Figure 12B). No acetone peak was observed based on 1H NMR spectroscopy (Figure 12C). Therefore, form L is likely anhydrous / hydrated.

[0165] Form L transforms into another form in a wet sample, and form L transforms into form I by storage at RT. Therefore, form L is presumed to be a metastable anhydrous / hydrated form that can be desolvated from the wet cake of the solvent system. Example 13A: Preparation of form M of compound 1 (form M)

[0166] Amorphous compound 1 (20 mg) was suspended in CHCl3 / n-heptane (1:1, v / v). The suspension was circulated from 50°C to 5°C to obtain form M.

[0167] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form M, indicating that form M is crystalline, see Figure 13A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 13A. [Table 13A]. XRPD pattern of form M of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 5.37 16.47 100.00 7.13 12.39 63.54 8.52 10.38 59.43 10.76 8.23 24.08 11.50 7.69 10.18 12.30 7.20 21.26 14.24 6.22 53.69 14.77 6.00 89.84 16.92 5.24 10.90 17.64 5.03 12.50 18.43 4.81 18.15 19.16 4.63 54.84 20.63 4.31 7.01 21.60 4.11 51.33 23.42 3.80 13.32

[0168] TGA / DSC curves showed a 1.6% weight loss at up to 170°C, and an endothermic peak at 171.0°C (peak) before decomposition (Figure 13B). Based on 1H NMR results, no obvious CHCl3 signal was observed (Figure 13C). VT-XPRD results showed no change in form after heating form M to 120°C in N2 and cooling it back to 30°C, indicating that form M is an anhydrous compound. Example 14A: Preparation of form N of compound 1 (form N)

[0169] Amorphous compound 1 (20 mg) was suspended in 0.5 mL ACN and stirred at 50°C to obtain form N.

[0170] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form N, indicating that form N is in a crystalline form, see Figure 14A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 14A. [Table 14A]. XRPD pattern of form N of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.73 13.13 100.00 9.32 9.49 8.75 11.53 7.67 6.33 12.37 7.16 7.10 13.15 6.73 5.35 13.59 6.52 3.45 14.32 6.19 15.35 15.07 5.88 23.16 15.52 5.71 12.47 16.11 5.50 6.41 16.53 5.36 3.46 16.94 5.23 2.46 18.00 4.93 18.18 19.52 4.55 6.27 20.22 4.39 8.03 20.69 4.29 8.63 21.43 4.15 13.04 21.98 4.04 4.63 22.49 3.95 3.42 23.59 3.77 1.73 24.29 3.66 1.84 26.46 3.37 1.24 27.07 3.29 2.28 28.31 3.15 1.80

[0171] TGA / DSC curves showed a weight loss of 0.3% at up to 160°C, and an endothermic peak at 160.6°C before decomposition (Figure 14B). 1H NMR results showed no ACN signal (Figure 14C). Combining the TGA and 1H NMR data, it is inferred that form N is an anhydrous form. Example 15A: Preparation of form O of compound 1 (form O)

[0172] Amorphous compound 1 (20 mg) was suspended in 0.5 mL of toluene at 50°C and stirred at 50°C to obtain form O.

[0173] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form O, indicating that form O is in a crystalline form, see Figure 15A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 15A. [Table 15A]. XRPD pattern of form O of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.90 12.81 100.00 8.50 10.41 9.61 10.88 8.13 53.19 12.37 7.16 52.85 13.14 6.74 11.30 13.55 6.54 21.25 13.83 6.40 70.34 14.80 5.99 32.57 15.17 5.84 22.34 15.88 5.58 13.21 16.44 5.39 8.86 16.88 5.25 68.94 17.07 5.20 68.37 18.27 4.86 19.15 18.97 4.68 55.62 19.27 4.61 13.03 19.49 4.55 12.56 19.92 4.46 11.93 20.33 4.37 8.46 21.00 4.23 40.76 21.91 4.06 19.76 22.23 4.00 18.01 23.05 3.86 6.26 23.71 3.75 9.70 24.04 3.70 9.97 24.54 3.63 4.45 25.06 3.55 22.88 25.62 3.48 9.59 26.48 3.37 4.21 26.98 3.31 8.87 27.65 3.23 7.28 29.72 3.01 6.34

[0174] TGA / DSC curves showed an 8.9% weight loss at temperatures up to 160°C, and four endothermic peaks at 115.8°C, 117.7°C, 146.6°C, and 175.8°C (Figure 15B) before decomposition. In the 1H NMR spectrum (Figure 15C), a toluene peak was observed, and the theoretical weight loss was determined to be 11.0%. The higher theoretical weight loss is likely due to inhomogeneous solvent residue. Heating experiments indicated that form O converted to form B after heating to 160°C and cooling back to RT. Combining the TGA and 1H NMR data, it is speculated that form O is a toluene solvate. Example 16A: Preparation of form P of compound 1 (form P)

[0175] Compound 1 (20 mg) was dissolved in chlorobenzene and centrifuged. The supernatant was exposed to toluene at RT to obtain form P.

[0176] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form P, indicating that form P is crystalline, see Figure 16A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 16A. [Table 16A]. XRPD pattern of form P of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.68 13.23 58.51 7.02 12.60 21.55 8.19 10.80 100.00 9.73 9.09 6.04 11.01 8.03 10.11 11.44 7.74 26.79 13.17 6.72 80.77 13.53 6.54 27.66 14.04 6.31 9.60 15.59 5.68 9.22 16.29 5.44 13.17 17.17 5.16 31.50 17.59 5.04 12.69 18.51 4.79 19.35 19.18 4.63 24.97 19.39 4.58 28.13 19.90 4.46 15.46 21.19 4.19 20.11 24.37 3.65 15.17

[0177] TGA / DSC results showed a 9.9% weight loss at up to 140°C, and an endothermic peak at 121.6°C (peak) before decomposition (Figure 16B). Based on the 1H NMR results (Figure 16C), the theoretical weight of chlorobenzene was calculated to be 9.8%, which matches the TGA weight loss. XRPD comparison showed that some diffraction peaks disappeared after ~4 weeks of RT storage. Heating the sample to 140°C resulted in the disappearance of more diffraction peaks. Combining the TGA, 1H NMR data, and heating experiments, form P is presumed to be a chlorobenzene solvent compound. Example 17A: Preparation of form Q of compound 1 (form Q)

[0178] Amorphous compound 1 (20 mg) was subjected to solid vapor diffusion in 1,4-dioxanone for 10 days at RT to obtain form Q.

[0179] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form Q, indicating that form Q is crystalline, see Figure 17A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 17A. [Table 17A]. XRPD pattern of form Q of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.89 12.83 100.00 10.31 8.58 0.85 11.90 7.44 1.19 13.76 6.44 2.12 15.28 5.80 0.48 17.22 5.15 3.72 18.44 4.81 2.30 20.07 4.42 4.18 21.74 4.09 0.74 22.54 3.94 0.80 23.37 3.81 0.67 24.91 3.57 0.18 25.79 3.45 0.23

[0180] TGA / DSC showed a 9.0% weight loss at up to 160°C, and an endothermic peak at 155.1°C (peak) before decomposition (Figure 17B). A 1,4-dichlorobenzene peak with a theoretical weight of 5.6% was detected in the 1H NMR spectrum (Figure 17C). The theoretical weight loss is lower than the TGA weight loss, likely due to solvent loss during storage. Form Q is presumed to be a 1,4-dichlorobenzene solvate. Example 18A: Preparation of Form R of Compound 1 (Form R)

[0181] Amorphous compound 1 (about 100 mg) was suspended in 0.5 mL ACN to obtain form R.

[0182] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form R, indicating that form R is in a crystalline form, see Figure 18A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 18A. [Table 18A]. XRPD patterns of form R of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.33 13.97 10.48 7.71 11.47 100.00 8.72 10.14 6.79 9.95 8.89 30.90 10.65 8.30 3.13 12.19 7.26 6.78 12.65 7.00 5.42 13.25 6.68 7.15 14.00 6.33 12.62 14.84 5.97 20.21 15.39 5.76 44.58 16.26 5.45 11.53 17.14 5.17 9.11 17.62 5.03 14.23 18.12 4.90 26.07 18.63 4.76 15.33 18.87 4.70 15.19 20.02 4.44 13.86 20.54 4.32 12.96 20.88 4.26 8.89 21.58 4.12 4.86 22.57 3.94 11.69 23.41 3.80 4.88 24.16 3.68 5.51 25.37 3.51 5.98 28.20 3.16 3.92

[0183] TGA / DSC curves showed a 2.8% weight loss at up to 120°C, and five endothermic peaks at 74.6°C, 89.5°C, 111.2°C, 130.0°C, and 168.6°C (peaks) before decomposition, and one exothermic peak at 144.6°C (Figure 18B). No ACN signal was observed in the 1H NMR spectrum (Figure 18C). VT-XRPD showed no change in form R after drying with N2 for 20 min; additional peaks and a significant peak shift were observed after heating form R to 100°C under N2 and cooling it back to 30°C. Considering the complex thermal signal observed in DSC before 100°C, form R is presumed to be anhydrous / hydrated. Example 19A: Preparation of form S of compound 1 (form S)

[0184] Compound 1 form R was heated to 150°C under a N2 atmosphere and then cooled back to 30°C to obtain form S.

[0185] X-ray powder diffraction (XRPD) patterns were used to characterize form S, indicating that form S is crystalline, see Figure 19A. Characteristic peaks and peak intensity percentages obtained from the XRPD analysis are listed in Table 19A. [Table 19A]. XRPD pattern of form S of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 5.84 15.14 15.29 7.24 12.22 100.00 9.38 9.43 37.96 11.53 7.67 49.13 11.88 7.45 32.48 14.01 6.32 61.09 14.69 6.03 75.74 14.98 5.91 27.89 15.40 5.76 36.26 16.30 5.44 30.21 17.23 5.15 47.74 17.54 5.06 37.46 17.96 4.94 76.34 18.41 4.82 16.47 19.41 4.57 29.43 19.74 4.50 20.80 20.09 4.42 30.41 20.46 4.34 37.54 21.10 4.21 18.10 21.57 4.12 18.65 22.35 3.98 18.57 22.74 3.91 15.87 23.31 3.82 15.84 24.65 3.61 11.24 24.99 3.56 22.96 25.76 3.46 4.45 26.57 3.35 3.45 27.39 3.26 10.16 28.38 3.14 9.01 28.98 3.08 3.76

[0186] TGA / DSC curves showed a 1.7% weight loss up to 120°C, and two endothermic peaks at 93.8°C and 169.5°C before decomposition (Figure 19B). Example 20A: Preparation of compound 1 in form T (form T)

[0187] Form N is converted to form T after being kept at 25°C / 60% RH and 40°C / 75% RH for one week and then sealed at 80°C for 24 hours. However, after being stored under the same conditions for 3 days, form T is converted back to form N.

[0188] X-ray powder diffraction (XRPD) patterns were used to characterize the obtained form T, indicating that form T is crystalline, see Figure 20A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 20A. [Table 20A]. XRPD pattern of form T of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.20 14.25 10.69 6.80 13.00 100.00 8.86 9.98 8.80 9.55 9.26 10.52 11.46 7.72 5.34 12.42 7.13 8.73 13.72 6.45 25.47 14.53 6.10 13.02 15.19 5.83 25.93 15.47 5.73 14.73 16.81 5.28 6.96 17.28 5.13 20.61 18.00 4.93 6.69 19.21 4.62 6.85 19.89 4.46 7.13 20.53 4.33 7.63 20.95 4.24 13.74 21.43 4.15 8.46 22.01 4.04 14.71 22.88 3.89 7.58 23.37 3.81 5.11 23.65 3.76 4.84 24.36 3.65 1.30 25.74 3.46 5.19 26.19 3.40 1.32 27.48 3.25 1.72 28.49 3.13 3.91

[0189] The XRPD coverage plot of this transformation shows that form T may be anhydrous / hydrated. Example 21A: Preparation of form U of compound 1 (form U)

[0190] At 20°C–40°C, n-heptane (140 mL) was slowly added to a solution of compound 1 (40 g) in DCM (240 mL). After stirring for 1 hour, another batch of n-heptane (20 mL) was added, and the mixture was kept at 20°C–40°C for 0.5 hours. This process was repeated with the addition of n-heptane (20 mL) for 0.5 hours, followed by the addition of n-heptane (20 mL). Finally, n-heptane (40 mL) was added and the mixture was stirred at 20°C–40°C for 12 hours. The mixture was filtered, and the resulting filter cake was dried at 45°C–55°C for 18 hours to give compound 1 in form U (36.5 g), which can be used as seed crystals.

[0191] At 25°C–35°C, n-heptane (14 kg) and seed crystals (0.020 kg) were added to a solution of compound 1 (6.6 kg) in DCM (51 kg). The mixture was stirred at 20°C–35°C for about 4.5 hours, and four batches of n-heptane (2.0 kg + 2.0 kg + 4.0 kg + 5.0 kg) were slowly added to the mixture, and then stirred separately at 20°C–35°C for about 2 hours. The mixture was then stirred at 20°C–35°C for about 16 hours. The mixture was filtered and washed with n-heptane (13 kg), and the resulting filter cake was dried at 45°C–55°C for 30 hours to give compound 1 in form U (5.86 kg) as a yellow solid.

[0192] X-ray powder diffraction (XRPD) plot (performed on a Bruker D8 advanced X-ray powder diffractometer) was used to characterize the obtained form U, indicating that form U is crystalline, see Figure 21A. Characteristic peaks and peak intensity percentages obtained from XRPD analysis are listed in Table 21A. [Table 21A]. XRPD plot of form U of compound 1 Position [°2θ] Interplanar spacing [Å] Relative strength [%) 6.968 12.67611 100.0 9.438 9.36287 13.1 10.237 8.63383 4.0 10.745 8.22714 4.0 11.282 7.83632 48.2 13.497 6.55533 53.3 13.920 6.35675 3.3 14.949 5.92133 8.4 15.019 5.89418 4.2 15.553 5.69298 32.7 16.073 5.50983 15.4 16.993 5.21352 16.6 17.116 5.17632 15.5 17.484 5.06834 13.1 18.036 4.91445 1.7 18.441 4.80742 1.7 18.908 4.68974 7.7 19.223 4.61343 2.2 19.549 4.53719 10.0 19.970 4.44261 13.9 20.515 4.32585 5.6 21.192 4.18916 13.9 21.613 4.10833 3.7 22.333 3.97756 2.0 22.600 3.93122 2.2 22.935 3.87453 2.0 23.633 3.76165 4.4 24.299 3.66000 9.0 25.736 3.45878 3.7 25.782 3.45273 4.4 26.147 3.40536 1.2 27.608 3.22838 2.6 28.462 3.13347 2.4 28.879 3.08910 1.0 29.317 3.04393 1.2

[0193] As shown in the TGA / DSC curves (performed on a NETZSCH TG 209 F1 instrument and TA DSC 250), a weight loss of 0.2% was detected up to 150°C, and an endothermic peak was observed at 170.8°C (Figures 21B and 21C). No DCM signal was detected in the 1H NMR spectrum (Figure 21D).

[0194] The DVS (Method B) cycle was carried out at 25°C. The adsorption and desorption system was correctable throughout the DVS cycle. The water absorption rate was 1.4% at 95% RH humidity. Compound 1 in form U has slight hygroscopicity.

[0195] The synthesis of compound 1, as shown in International Patent Publication WO 2019 / 210828, involves reacting the acidic intermediate (S)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(2-(2-(2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzoic acid with the sulfonamide intermediate 4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrobenzenesulfonamide to obtain compound 1. Simultaneously, due to the reaction of the azaindole moiety in compound 1 with the acidic intermediate, a dimer compound is generated, becoming a process impurity. In manufacturing, only form U can unexpectedly and effectively remove the dimer impurity. In one manufacturing batch, the dimer impurity content in the process control (IPC) was 0.4% (wt); after EA crystallization, form A was obtained, with the dimer impurity content still at 0.4%; further, after recrystallization with a THF / ACN mixed solution, the dimer impurity content decreased to 0.22%; finally, after recrystallization with a DCM / heptane mixed solution, form U was obtained, and no dimer impurities were detected. Example 21B: Preparation of form U of compound 1

[0196] At RT, amorphous compound 1 (20 mg) was suspended in a mixture of DCM / n-heptane (1:1, v / v). The suspension was stirred at RT for 1–7 days to slurry up to obtain form U. Example 21C: Preparation of form U of compound 1

[0197] Compound 1 (2.0 g) was dissolved in DCM (20 mL) at 40°C. Heptane (15 mL) was added to the solution and stirred at 40°C, followed by the addition of heptane (5.0 mL) at 40°C. The mixture was cooled to RT and stirred to produce a precipitate. The precipitate was filtered, washed with heptane, and dried to give the product (1.4 g). Example 22A: Compound 1, amorphous form (amorphous form)

[0198] Compound 1 (153.5 g) was dissolved in DCM (1.0 L) to obtain a clear solution. The solution was concentrated under reduced pressure to remove the solvent, and the residue was mixed with MTBE (1.0 L) to form a slurry and filtered. The filter cake was collected and dried under vacuum to give the product (137.5 g).

[0199] The obtained amorphous form is shown in the X-ray powder diffraction (XRPD) pattern of Figure 22A. TGA / DSC (Figure 22B) results show weight loss in two phases (0.7% up to 110°C and 0.5% from 110°C to 200°C) and a possible glass transition signal (moderate) at 126.7°C. Chemical purity was determined to be 98.3% by high-performance liquid chromatography (HPLC). DVS results indicate a water absorption rate of 1.8% at 80% RH / 25°C. Physical stability

[0200] To assess physicochemical stability, 1–3 mg of samples in the forms of B, S, M, R, F, H, and N were stored at 25°C / 60% RH or 40°C / 75% RH for one week (unsealed) or at 80°C for 24 hours (sealed).

[0201] The XRPD overlay plot showed no changes in the forms B, S, and M.

[0202] No change in form R was observed after one week of storage at 25°C / 60% RH or 40°C / 75% RH. After 24 hours of storage at 80°C / sealed, form R transformed into a form similar to form S.

[0203] For form F, as shown in the XRPD coverage plot, a mixture of form F and B was observed after storing the form F sample under all test conditions.

[0204] The XRPD overlay plot shows that no change in the form of form H was observed after one week of continuous exposure at 25°C / 60% RH or 40°C / 75% RH, but a significant decrease in crystallinity was observed after exposure to 80°C / sealed for 24 hours.

[0205] Form N is converted to form T, and form T may convert back to form N when stored at RT for ~3 days. Solid form solubility.

[0206] The solubility of compound 1 in different physical forms was tested in water, 0.1 N HCl, pH 4.5 acetate buffer, and pH 6.8 phosphate buffer. The concentration of compound 1 was determined by HPLC at 24-hour time points.

[0207] For the amorphous form of compound 1, compound 1 was not detected in water, pH 4.5 buffer, and pH 6.8 buffer, but the corresponding solubility in 0.1 N HCl was 35.30 µg / mL. For form A of compound 1, the corresponding solubilities in water, pH 4.5, and pH 6.8 buffer were 0.37 µg / mL, 0.76 µg / mL, and 0.43 µg / mL, respectively, but the corresponding solubility in 0.1 N HCl was 29.36 µg / mL. Therefore, form A showed higher solubility in water, pH 4.5, and pH 6.8 buffer, and lower solubility in 0.1 N HCl compared to the amorphous form. Solid form stability

[0208] The solubility of form B in solid form was evaluated in an acetone / H₂O system at 50°C. Approximately 2 mg of form B sample was slurried or shaken in acetone / H₂O (1:9, v / v) and H₂O solutions (by saturating the amorphous sample).

[0209] The crystallization state under the slurry state was tracked. The XRPD overlay plot showed that: 1) After slurrying form B in acetone / H2O (1:9, v / v) or H2O for about 4 days, a decrease in crystallinity was observed in form B (including amorphous form); and 2) After slurrying form B in H2O for about 4 hours, no change in form was observed.

[0210] In addition, the crystallization state under oscillation was also monitored. XRPD coverage plots showed that no change in form B was observed after oscillation in acetone / H₂O (1:9, v / v) or H₂O for approximately 4 hours or 4 days, suggesting that form B may be affected by mechanical forces. Physical and chemical stability tests

[0211] Long-term and accelerated stability studies of different physical forms of compound 1 were conducted by storing samples at 25 ± 2°C / 60 ± 5% RH and 40 ± 2°C / 75 ± 5% RH for up to 6 months. The total impurity content of each sample was determined by HPLC.

[0212] For the amorphous form of compound 1, the chemical purity of compound 1 is significantly reduced, for example, when stored at 40 ± 2°C / 75 ± 5% RH for 6 months, the total impurity content increases from 2.1% to 4.2%, and many new impurities are detected.

[0213] For compound 1 form A, the chemical purity of compound 1 changed significantly. For example, when stored at 40 ± 2°C / 75 ± 5% RH for 6 months, the total impurity content increased only from 0.40% to 0.52%. In addition, no changes in crystalline form and optical purity were observed, but the content of solvent EA decreased slightly from about 9.5 to 8.8 (x10 4 ppm).

[0214] For compound 1 in form U, the chemical purity of compound 1 changed significantly. For example, when stored at 40 ± 2°C / 75 ± 5% RH for 6 months, the total impurity content increased only from 0.40% to 0.72%. In addition, no changes in crystalline form or optical purity were observed.

[0215] Therefore, both form A and form U of compound 1 exhibit better physical stability than its amorphous form, and form A exhibits better chemical stability.

[0216] Although the present invention has been described in conjunction with its specific embodiments, it should be understood that it is capable of further modifications, and this application is intended to cover any variations, uses or adaptations of the invention that generally follow the principles of the invention and are included within the scope of known or customary practice in the field to which the invention pertains and that are applicable to the essential features described above and deviations from this disclosure within the scope of the appended claims. [Simplified Explanation of the Diagram]

[0084] Figure 1A shows the X-ray powder diffraction (XRPD) pattern of compound form A (EtOAc solvate 1:1) prepared according to Example 1A. Figure 1B shows the differential scanning calorimetry (DSC) curve of compound form A prepared according to Example 1A. Figure 1C shows the thermogravimetric analysis (TGA) curve of compound form A prepared according to Example 1A. Figure 1D shows the 1H-NMR spectrum of compound form A (EtOAc solvate 1:1) prepared according to Example 1A. Figure 1E shows the calculated XRPD of the single crystal structure and the experimental XRPD of the single crystal of compound form A. Figure 2A shows the X-ray powder diffraction (XRPD) pattern of compound form B (anhydrous) prepared according to Example 2A. Figure 2B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form B prepared according to Example 2A. Figure 2C shows the 1H-NMR spectrum of compound form B prepared according to Example 2A. Figure 2D shows the XRPD coverage of compound form B prepared according to Example 2A before heating, heating to 120°C, and heating to 160°C. Figure 3A shows the X-ray powder diffraction (XRPD) pattern of compound form C (MEK solvate) prepared according to Example 3A. Figure 3B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form C prepared according to Example 3A. Figure 3C shows the 1H-NMR spectrum of compound form C prepared according to Example 3A. Figure 4A shows the X-ray powder diffraction (XRPD) pattern of compound form D (IPAc solvate) prepared according to Example 4A. Figure 4B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form D prepared according to Example 4A. Figure 4C shows the 1H-NMR spectrum of compound form D prepared according to Example 4A. Figure 5A shows the X-ray powder diffraction (XRPD) pattern of compound form E (anisole solvate) prepared according to Example 5A. Figure 5B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form E prepared according to Example 5A. Figure 5C shows the 1H-NMR spectrum of compound form E prepared according to Example 5A. Figure 6A shows the X-ray powder diffraction (XRPD) pattern of compound form F prepared according to Example 6A. Figure 6B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form F prepared according to Example 6A.Figure 6C shows the 1H-NMR spectrum of compound form F prepared according to Example 6A. Figure 7A shows the X-ray powder diffraction (XRPD) pattern of compound form G prepared according to Example 7A. Figure 7B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form G prepared according to Example 7A. Figure 7C shows the 1H-NMR spectrum of compound form G prepared according to Example 7A. Figure 8A shows the X-ray powder diffraction (XRPD) pattern of compound form H (anhydrous / hydrated) prepared according to Example 8A. Figure 8B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form H prepared according to Example 8A. Figure 8C shows the 1H-NMR spectrum of compound form H prepared according to Example 8A. Figure 9A shows the X-ray powder diffraction (XRPD) pattern of compound form I (IPA solvate) prepared according to Example 9A. Figure 9B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form I prepared according to Example 9A. Figure 9C shows the 1H-NMR spectrum of compound form I prepared according to Example 9A. Figure 9D shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form I prepared according to Example 9B. Figure 9E shows the 1H-NMR spectrum of compound form I prepared according to Example 9B. Figure 10A shows the X-ray powder diffraction (XRPD) pattern of compound form J (2-MeTHF solvate) prepared according to Example 10A. Figure 10B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form J prepared according to Example 10A. Figure 10C shows the 1H-NMR spectrum of compound form J prepared according to Example 10A. Figure 11A shows the X-ray powder diffraction (XRPD) pattern of compound form K (methyl acetate solvate) prepared according to Example 11A. Figure 11B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form K prepared according to Example 11A. Figure 11C shows the 1H-NMR spectrum of compound form K prepared according to Example 11A. Figure 12A illustrates the X-ray powder diffraction (XRPD) pattern of compound form L (anhydrous / hydrated) prepared according to Example 12A.Figure 12B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form L prepared according to Example 12A. Figure 12C shows the 1H-NMR spectrum of compound form L prepared according to Example 12A. Figure 13A illustrates the X-ray powder diffraction (XRPD) pattern of compound form M (anhydrous) prepared according to Example 13A. Figure 13B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form M prepared according to Example 13A. Figure 13C shows the 1H-NMR spectrum of compound form M prepared according to Example 13A. Figure 14A shows the X-ray powder diffraction (XRPD) pattern of compound form N (anhydrous) prepared according to Example 14A. Figure 14B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curves of compound form N prepared according to Example 14A. Figure 14C shows the 1H-NMR spectrum of compound form N prepared according to Example 14A. Figure 15A shows the X-ray powder diffraction (XRPD) pattern of compound form O (toluene solvate) prepared according to Example 15A. Figure 15B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form O prepared according to Example 15A. Figure 15C shows the 1H-NMR spectrum of compound form O prepared according to Example 15A. Figure 16A shows the X-ray powder diffraction (XRPD) pattern of compound form P (chlorobenzene solvate) prepared according to Example 16A. Figure 16B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form P prepared according to Example 16A. Figure 16C shows the 1H-NMR spectrum of compound form Q prepared according to Example 16A. Figure 17A shows the X-ray powder diffraction (XRPD) pattern of compound form Q (1,4-dichlorodimethylamine solvate) prepared according to Example 17A. Figure 17B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form Q prepared according to Example 17A. Figure 17C shows the 1H-NMR spectrum of compound form Q prepared according to Example 17A. Figure 18A shows the X-ray powder diffraction (XRPD) pattern of compound form R (anhydrous / hydrated) prepared according to Example 18A. Figure 18B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form R prepared according to Example 18A.Figure 18C shows the 1H-NMR spectrum of compound form R prepared according to Example 18A. Figure 19A shows the X-ray powder diffraction (XRPD) pattern of compound form S prepared according to Example 19A. Figure 19B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound form S prepared according to Example 19A. Figure 19C shows the 1H-NMR spectrum of compound form S prepared according to Example 19A. Figure 20A shows the X-ray powder diffraction (XRPD) pattern of compound form T prepared according to Example 19A. Figure 21A shows the X-ray powder diffraction (XRPD) pattern of compound form U (anhydrous) prepared according to Example 21A. Figure 21B shows the differential scanning calorimetry (DSC) curve of compound form U prepared according to Example 21A. Figure 21C shows the thermogravimetric analysis (TGA) curve of compound 1 in form U prepared according to Example 21A. Figure 21D shows the 1H-NMR spectrum of compound 1 in form U prepared according to Example 21. Figure 22A shows the X-ray powder diffraction (XRPD) pattern of the amorphous form of compound 1. Figure 22B shows the differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) curve of compound 1 in its amorphous form. Figure 23 shows the interconversion of the crystalline forms of compound 1.

Claims

1. A crystalline form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzylamine (compound 1), which is an EtOAc solvate, each mole of which contains about 1 mole of EtOAc, said form being designated as form A, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 16.5 ± 0.1° and 24.5 ± 0.1°.

2. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 12.4 ± 0.1°, 16.5 ± 0.1° and 24.5 ± 0.1°.

3. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 12.4 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

4. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

5. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

6. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 20.7 ± 0.1° and 24.5 ± 0.1°.

7. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 20.7 ± 0.1°, and 24.5 ± 0.1°.

8. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.7 ± 0.1°, and 24.5 ± 0.1°.

9. The crystalline form as described in claim 1, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.9 ± 0.1°, 10.6 ± 0.1°, 12.4 ± 0.1°, 13.8 ± 0.1°, 14.1 ± 0.1°, 16.5 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.7 ± 0.1°, and 24.5 ± 0.1°.

10. The crystalline form as described in claim 1, wherein form A has an XRPD diagram substantially as shown in Figure 1A or Figure 1E.

11. The crystalline form as described in claim 1, wherein form A is characterized by having two endothermic peaks at about 150°C and about 178°C according to differential scanning calorimetry (DSC).

12. The crystalline form as described in claim 1, wherein form A has a DSC thermogram substantially as shown in Figure 1B.

13. The crystal form as claimed in claim 1, wherein form A is a triclinic crystal system and has a space group of P1, having the following cell parameters: (a) about 13.644 Å, (b) about 14.070 Å, (c) about 15.012 Å, (α) about 112.0202(3)°, (β) about 104.6821(3)°, and (γ) about 93.6507(2)°.

14. A crystalline form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzylamine (compound 1), wherein the form is anhydrous, the form being designated as form B, wherein the X-ray powder diffraction pattern of the crystalline form has a diffraction peak at an angle of 14.4 ± 0.1° at 2θ.

15. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1° and 17.5 ± 0.1°.

16. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1°, 17.5 ± 0.1° and 18.4 ± 0.1°.

17. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

18. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at °2θ values ​​independently selected from the group consisting of: 7.2 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1°, and 19.6 ± 0.1°.

19. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.7 ± 0.1°, 7.2 ± 0.1°, 13.8 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

20. The crystalline form as described in claim 14, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 6.7 ± 0.1°, 7.2 ± 0.1°, 13.8 ± 0.1°, 14.4 ± 0.1°, 17.5 ± 0.1°, 18.4 ± 0.1° and 19.6 ± 0.1°.

21. The crystalline form as described in claim 14, wherein form B has an XRPD diagram substantially as shown in Figure 2A or Figure 2D.

22. The crystalline form as described in claim 14, wherein form B is characterized by having two endothermic peaks at approximately 107.7°C and 187°C according to differential scanning calorimetry (DSC).

23. The crystalline form as described in claim 14, wherein form B has a DSC thermogram substantially as shown in Figure 2B.

24. A crystalline form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzylamine, wherein the form is anhydrous, the form being designated as form U, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1° and 24.3 ± 0.1°.

25. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 15.6 ± 0.1° and 24.3 ± 0.1°.

26. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 15.6 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

27. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

28. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at °2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

29. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 21.2 ± 0.1° and 24.3 ± 0.1°.

30. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

31. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

32. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

33. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 17.0 ± 0.1°, 17.5 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, and 24.3 ± 0.1°.

34. The crystalline form as described in claim 24, wherein the X-ray powder diffraction pattern of the crystalline form has diffraction peaks at 2θ angles of 7.0 ± 0.1°, 9.4 ± 0.1°, 11.3 ± 0.1°, 13.5 ± 0.1°, 15.6 ± 0.1°, 16.1 ± 0.1°, 17.0 ± 0.1°, 17.5 ± 0.1°, 19.5 ± 0.1°, 20.0 ± 0.1°, 21.2 ± 0.1°, 21.6 ± 0.1°, and 24.3 ± 0.1°.

35. The crystalline form as described in claim 24, wherein form U has an XRPD diagram substantially as shown in FIG21A.

36. The crystalline form as described in claim 24, wherein form U is characterized by having an endothermic peak at approximately 171°C according to differential scanning calorimetry (DSC).

37. The crystalline form as described in claim 24, wherein form U has a DSC thermogram substantially as shown in Figure 21B.

38. An amorphous form of 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-N-((4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrophenyl)sulfonyl)-4-(2-((S)-2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonane-7-yl)benzamide (compound 1).

39. A pharmaceutical composition comprising (a) a therapeutically effective amount of the crystalline form as described in any one of claims 1-37, or the amorphous form of compound 1 as described in claim 38; and (b) one or more pharmaceutically acceptable excipients.

40. An use of a crystalline form as described in any one of claims 1-37, or an amorphous form of compound 1 as described in claim 38, or a pharmaceutical composition as described in claim 39, for the preparation of a medicament for treating diseases associated with Bcl-2 protein inhibition.

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