Solid forms of Bcl-2 inhibitors, methods for their preparation and use - Patents.com

Abstract

Disclosed are solid forms, particularly crystalline forms, 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]nonan-7-yl)benzamide, pharmaceutical compositions comprising said solid forms, processes for preparing said solid forms, and methods of their use.

Description

[Technical field]

[0001] Disclosed herein are solid forms 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]nonan-7-yl)benzamide, pharmaceutical compositions comprising said solid forms, processes for preparing said solid forms, and methods of their use. [Background technology]

[0002] Programmed cell death, or apoptosis, occurs in multicellular organisms to remove damaged or unnecessary cells and is crucial for normal tissue homeostasis (Br. J.Cancer 1972,26,239). However, defective apoptotic processes are involved in a variety of diseases. Excessive apoptosis leads to atrophy, while insufficient amounts of apoptosis lead to uncontrolled cell proliferation, e.g., cancer (Cell 2011,144,646). Resistance to apoptotic cell death is a hallmark of cancer and contributes to chemotherapy resistance (Nat Med.2004,10,789-799). In cancer, several major pathways controlling apoptosis are commonly altered. Some factors, such as the Fas receptor and caspases, promote apoptosis, while some members of the B-cell lymphoma 2 (Bcl-2) protein family inhibit apoptosis. Negative regulation of apoptosis inhibits cell death signaling pathways, helping tumors avoid cell death and resulting in drug resistance.

[0003] There are two distinct apoptotic pathways: the extrinsic pathway and the intrinsic pathway. The extrinsic pathway is activated in response to the binding of death-inducing ligands to cell surface death receptors (Nat Rev Drug Discov. 2017 16, 273-284). The B-cell lymphoma 2 (BCL-2) gene family, a group of proteins with homology to the Bcl-2 protein, encodes more than 20 proteins that control the intrinsic apoptotic pathway. Bcl-2 family proteins are characterized by containing at least one of the four conserved Bcl-2 homology (BH) domains (BH1, BH2, BH3, and BH4) (Nat.Rev.Cancer 2008, 8, 121; Mol.Cell 2010, 37, 299; Nat.Rev.Mol.Cell Biol. 2014, 15, 49). Bcl-2 family proteins consist of pro-apoptotic and anti-apoptotic molecules and can be classified into three subfamilies according to sequence homology with the four BH domains: (1) subfamilies that share sequence homology in all four BH domains, e.g., anti-apoptotic Bcl-2, Bcl-XL, and Bcl-w; (2) subfamilies that share sequence homology in BH1, BH2, and BH4, e.g., pro-apoptotic Bax and Bak; and (3) subfamilies that share sequence homology only in BH3, e.g., pro-apoptotic Bik, Bid, and HRK. One of the unique features of Bcl-2 family proteins is heterodimerization between anti-apoptotic and pro-apoptotic proteins, which is believed to inhibit the biological activity of the partner. This heterodimerization is mediated by the insertion of the BH3 region of the pro-apoptotic protein into a hydrophobic groove composed of BH1, BH2, and BH3 from the anti-apoptotic protein. For anti-apoptotic activity, the BH4 domain is required in addition to BH1 and BH2, whereas the BH3 domain is important and sufficient for pro-apoptotic activity.

[0004] Similar to oncogene addiction, where tumor cells depend on a single dominant gene to survive, tumor cells may also become dependent on Bcl-2 to survive.Bcl-2 overexpression is frequently seen in acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), relapsed / refractory chronic lymphocytic leukemia (CLL), follicular lymphoma (FL), non-Hodgkin's lymphoma (NHL), and solid tumors such as pancreatic cancer, prostate cancer, breast cancer, small cell lung cancer, and non-small cell lung cancer (Cancer 2001,92,1122-1129; Cancer Biol.2003;13:15-23; Curr.Cancer Drug Targets 2008,8,207-222; Cancers 2011,3,1527-1549). Dysregulation of apoptotic pathways has also been implicated in the pathology of other serious diseases, e.g., neurodegenerative conditions such as Alzheimer's disease (upregulated apoptosis), as well as proliferative diseases such as cancer, autoimmune diseases and prothrombotic conditions (downregulated apoptosis).

[0005] International Publication No. WO 2019 / 210828 discloses a series of Bcl-2 inhibitors, in particular 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]nonan-7-yl)benzamide (hereinafter Compound 1), which selectively inhibits Bcl-2 protein for the treatment of diseases of apoptosis dysregulation, such as cancer, autoimmune diseases and prothrombotic conditions.

[0006] Compound 1 has 13 freely rotatable bonds and a high molecular weight (Mw>800). Molecules with high conformational flexibility tend to be extremely difficult to crystallize, and the most important molecular descriptors responsible for the crystallization behavior of these molecules were 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, it is not possible to predict whether a pure physical form can be obtained for a particular compound, especially one with a high molecular weight and many freely rotatable bonds, and which physical form is stable and suitable for use in pharmaceutical products. Similarly, it is equally impossible to predict whether a particular crystalline solid-state physical form can be produced, with the desired chemical and physical properties suitable for pharmaceutical formulations. Summary of the Invention [Problem to be solved by the invention]

[0007] For all of the above reasons, there is a great need to find a crystalline form of Compound 1 that offers good stability and good ease of manufacture. The present disclosure advantageously meets one or more of these requirements. [Means for solving the problem]

[0008] The present disclosure addresses the above problems and needs by providing a solid form, preferably a crystalline form, of Compound 1, which is suitable for use in pharmaceuticals. Compound 1 was found to have multiple freely rotatable bonds and a high molecular weight of over 800, but the inventors of the present disclosure unexpectedly discovered 21 crystalline forms of Compound 1, namely, 6 anhydrates (forms B, S, U, M, F and N), 4 hydrates / anhydrates (forms H, R, L and T), and 11 solvates (forms A, C, D, E, G, I, J, K, O, P and Q). Form I isomorphous during formation, Form L is a metastable form, Forms N and T can be converted into each other during storage, and Form S was obtained by heating Form R to 150°C.

[0009] The inventors discovered that Form A was an EtOAc solvate of Compound 1. Form A has good physical properties, such as better physical stability and better solubility. However, it is difficult to control the ethyl acetate content of Form A during manufacturing, storage and formulation, and Form A may convert to Form B after Form A is heated to 160°C, cooled to room temperature and re-exposed to air atmosphere.

[0010] Solvate forms C, D, J, K and O and anhydrous form F can be converted to anhydrous form B after heating to high temperatures. Forms K and F can be converted spontaneously to form B after prolonged storage. Form R can be converted to anhydrous form S after heating to 150°C.

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

[0012] Furthermore, Form B has good thermodynamic stability with a high melting point and slight hygroscopicity with a water uptake of 0.9% at 25° C. / 80% RH. Form B also exhibited good physicochemical and thermodynamic stability after exposure under 25° C. / 80% RH and shaking in acetone / HO (1:9, v / v) and HO for about 4 days.

[0013] We tried to scale up form B, but failed to obtain the desired form directly by normal crystallization. Form A had to be heated or form K had to be treated in a specific solvent at a temperature of about 100°C to obtain form B, which could not meet the requirements of the scale-up process. Form M with good stability was obtained as an anhydrous form from a solvent of CHCl3 and heptane, but CHCl3 is not environmentally friendly and belongs to Class 2 with a low ICH guideline acceptable daily exposure (PDE) of 0.6 mg / day. Form U of compound 1 as an anhydrous form was unexpectedly obtained by replacing CHCl3 with DCM in the recrystallization process, which is reproducible and suitable for the scale-up process. Form U showed good physicochemical, thermodynamic and physical stability. For example, no significant change in chemical purity, crystal form and optical purity occurred when stored at 25±2°C / 60±5% RH or 40±2°C / 75±5% RH for up to 6 months. Furthermore, only Form U can remove the important dimeric impurity in the preparation, which is a process impurity that is essentially formed by the reaction of compound 1 with the acid intermediate (S)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(2-(2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonan-7-yl)benzoic acid.

[0014] Although Form U has a lower melting point than Form B, Form U does not have the challenges from, for example, preparation, scale-up process, solvent residues, and API and pharmaceutical formulation qualification issues, and has good stability and formation performance through solution precipitation. Therefore, Form U is more suitable for manufacturing and pharmaceutical formulation.

[0015] In a first aspect, disclosed herein is 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]nonan-7-yl)benzamide, designated herein as Form A.

[0016] In a second aspect, disclosed herein is a crystalline form of Compound 1, which is an EtOAc solvate containing about 1 mole of EtOAc per mole.

[0017] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 16.5±0.1° and 24.5±0.1°.

[0018] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 12.4±0.1°, 16.5±0.1°, and 24.5±0.1°.

[0019] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 12.4±0.1°, 16.5±0.1°, 20.7±0.1°, and 24.5±0.1°.

[0020] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form is at 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°, 19.2±0.1°, 20.2±0.1°, 21.2±0.1°, 22.2±0.1°, 23.2±0.1°, 24.2±0.1°, 25.2±0.1°, 26.2±0.1°, 27.2±0.1°, 28.2±0.1°, 29.2±0.1°, 30.2±0.1°, 31.2±0.1°, 32.2±0.1°, 33.2±0.1°, 34.2±0.1°, 35.2±0.1°, 36.2±0.1°, 37.2±0.1°, 38.2±0.1°, 39.2±0.1°, 40.2±0.1°, 41.2±0.1°, 42.2±0.1°, 43.2±0.1°, 44.2±0.1°, 45.2±0.1 The compound has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at: 18.5±0.1°, 19.1±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 pattern substantially as shown in FIG. 1A or FIG. 1E.

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

[0029] In some embodiments, Form A has a DSC thermogram substantially as shown in FIG. 1B.

[0030] In some embodiments, Form A is characterized by a triclinic crystal system and space group P1 with lattice parameters of (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 third aspect, disclosed herein is a crystalline form of Compound 1, designated Form B, which is anhydrous.

[0032] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising a diffraction peak having a °2θ angle value at 14.4±0.1°.

[0033] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1° and 17.5±0.1°.

[0034] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1°, 17.5±0.1°, and 18.4±0.1°.

[0035] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1°, 17.5±0.1°, 18.4±0.1°, and 19.6±0.1°.

[0036] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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, Form B has an XRPD pattern substantially as shown in FIG. 2A or FIG. 2D.

[0041] In some embodiments, Form B is characterized by having one endothermic peak at about 187° C. by differential scanning calorimetry (DSC).

[0042] In some embodiments, Form B has a DSC thermogram substantially as shown in Figure 2B.

[0043] In a fourth aspect, disclosed herein is a crystalline form of Compound 1, designated Form U, which is anhydrous.

[0044] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 11.3±0.1° and 24.3±0.1°.

[0045] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 11.3±0.1°, 15.6±0.1°, and 24.3±0.1°.

[0046] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 11.3±0.1°, 15.6±0.1°, 21.2±0.1°, and 24.3±0.1°.

[0047] In some embodiments, the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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 crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having degree 2θ angle values ​​at 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°, and 24.3±0.1°.

[0055] In some embodiments, the crystalline form is 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. and 29.3±0.1°.

[0056] In some embodiments, form U has an XRPD pattern substantially as shown in FIG. 21A.

[0057] In some embodiments, Form U is characterized by having one endothermic peak at about 164° C. by differential scanning calorimetry (DSC).

[0058] In some embodiments, Form U has a DSC thermogram substantially as shown in FIG. 21B.

[0059] In a fifth aspect, the crystalline form of Compound 1 is designated 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, 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 and Form T have XRPD patterns substantially as shown in Figure 3A, Figure 4A, Figure 5A, Figure 6A, Figure 7A, Figure 8A, Figure 9A, Figure 10A, Figure 11A, Figure 12A, Figure 13A, Figure 14A, Figure 15A, Figure 16A, Figure 17, Figure 18A, Figure 19A or Figure 20A, respectively.

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

[0062] In a sixth aspect, disclosed herein is 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]nonan-7-yl)benzamide (Compound 1).

[0063] In some embodiments, an amorphous form of Compound 1 has an XRPD pattern substantially as shown in FIG. 22A.

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

[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 seventh aspect, disclosed herein is a pharmaceutical composition comprising (a) a therapeutically effective amount of a solid form of Compound 1, preferably a crystalline form of Compound 1 disclosed herein, or an amorphous form of Compound 1, and (b) one or more pharma- ceutically acceptable excipients.

[0067] In some embodiments, the crystalline form of Compound 1 is an EtOAc solvate of Compound 1 containing about 1 mole of EtOAc per mole, and an anhydrous crystalline 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 of Compound 1.

[0070] In an eighth aspect, disclosed herein is a process for preparing a pharmaceutical solution of Compound 1 comprising dissolving a solid form of Compound 1, preferably a crystalline form of Compound 1 as described in claim 1, in a pharma- ceutically acceptable solvent or mixture of solvents, or comprising an amorphous form of Compound 1.

[0071] In a ninth aspect, disclosed herein is a method for treating a disease associated with Bcl-2 protein inhibition, comprising administering to a subject a therapeutically effective amount of a crystalline form of Compound 1, an amorphous form of Compound 1, or a pharmaceutical composition disclosed herein.

[0072] In some embodiments, the disease associated with Bcl-2 protein inhibition is a disease of apoptosis dysregulation. In some preferred embodiments, the disease associated with Bcl-2 protein inhibition is a neoplastic disease, a prothrombotic disease, an immune disease, or an 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 an EtOAc solvate of Compound 1 containing about 1 mole of EtOAc per mole, or an anhydrous crystalline 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 of Compound 1.

[0076] In some embodiments, a therapeutically effective amount is administered orally at a dosage of about 1 mg to about 640 mg of Compound 1 per day.

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

[0078] In some embodiments, Form A is obtained by a process comprising any one of the following steps: a) Compound 1 is dissolved in DCM, the DCM is removed, and EA is charged to obtain Form A; b) Compound 1 is dissolved in DCM, concentrated, charged with EA, and the DCM is exchanged with EA, MeOH, and EA separately to obtain Form A; c) dissolving Compound 1 in EA, heating and cooling to obtain Form A; or d) Compound 1 is dissolved in a THF / EtOAc (1:2, v / v) solvent mixture and evaporated to obtain Form A.

[0079] In some embodiments, Form B is obtained by a process comprising any one of the following steps: a) dissolving compound 1 in acetone and evaporating the solvent to obtain the desired crystalline form; b) heating Form A, Form C, and Form O to about 160° C. and cooling to room temperature to obtain Form B; c) stepwise isothermal heating of Form A to about 100° C. to obtain Form B; d) heating Form D or Form J to about 130° C. and further isothermal to obtain Form B; or e) Form K is added to heptane and refluxed at about 100° C. and cooled to obtain Form B.

[0080] In some embodiments, Form U is obtained by a process comprising any one of the following procedures: a) Compound 1 is dissolved in DCM and the batch is added with n-heptane and stirred to obtain Form U; b) Compound 1 is dissolved in a mixture of DCM / n-heptane (1:1, v / v) and stirred to obtain Form U.

[0081] In some embodiments, Form A and / or Form B are obtained by a process comprising adding a crystalline seed into a solution system.

[0082] In some embodiments, the amorphous form is obtained by a process comprising any one of the following procedures: a) Compound 1 is dissolved in DCM and dried to obtain an amorphous form, or b) Compound 1 is dissolved in a mixture of solvents containing DCM and dried to obtain an amorphous form.

[0083] In some embodiments, the amorphous form is obtained by a process comprising dissolving Compound 1 in a solid form, preferably a crystalline form, of Compound 1. [Brief description of the drawings]

[0084] [Figure 1A]FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form A (1:1 EtOAc solvate) prepared according to Example 1A. [Figure 1B] 1 is a differential scanning calorimetry (DSC) profile of Compound 1 Form A prepared according to Example 1A. [Figure 1C] 1 shows a thermogravimetric analysis (TGA) profile of Compound 1 Form A prepared according to Example 1A. [Figure 1D] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of Compound 1 Form A (1:1 EtOAc solvate) prepared according to Example 1A. [Figure 1E] 1 shows the calculated and experimental XRPD of the single crystal structure of Compound 1 Form A. [Figure 2A] FIG. 2 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form B (anhydrous) prepared according to Example 2A. [Figure 2B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form B prepared according to Example 2A. [Figure 2C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form B prepared according to Example 2A. [Figure 2D] 1 shows XRPD overlay patterns of Compound 1 Form B prepared according to Example 2A before heating, heated to 120°, and heated to 160°. [Figure 3A] FIG. 2 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form C (MEK solvate) prepared according to Example 3A. [Figure 3B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form C prepared according to Example 3A. [Figure 3C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of Compound 1 Form C prepared according to Example 3A. [Figure 4A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form D (IPAc solvate) prepared according to Example 4A. [Figure 4B]1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form D prepared according to Example 4A. [Figure 4C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form D prepared according to Example 4A. [Figure 5A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form E (anisole solvate) prepared according to Example 5A. [Figure 5B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form E prepared according to Example 5A. [Figure 5C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form E prepared according to Example 5A. [Figure 6A] 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form F prepared according to Example 6A. [Figure 6B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form F prepared according to Example 6A. [Figure 6C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form F prepared according to Example 6A. [Figure 7A] 1 shows an X-ray powder diffraction (XRPD) pattern of Compound 1 Form G prepared according to Example 7A. [Figure 7B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form G prepared according to Example 7A. [Figure 7C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form G prepared according to Example 7A. [Figure 8A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form H (anhydrous / hydrate) prepared according to Example 8A. [Figure 8B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form H prepared according to Example 8A. [Figure 8C]1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form H prepared according to Example 8A. [Figure 9A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form I (IPA solvate) prepared according to Example 9A. [Figure 9B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form I prepared according to Example 9A. [Figure 9C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 Form I prepared according to Example 9A. [Figure 9D] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form I prepared according to Example 9B. [Figure 9E] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 Form I prepared according to Example 9B. [Figure 10A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form J (2-MeTHF solvate) prepared according to Example 10A. [Figure 10B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form J prepared according to Example 10A. [Figure 10C] FIG. 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form J prepared according to Example 10A. [Figure 11A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form K (methyl acetate solvate) prepared according to Example 11A. [Figure 11B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form K prepared according to Example 11A. [Figure 11C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form K prepared according to Example 11A. [Figure 12A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form L (anhydrous / hydrate) prepared according to Example 12A. [Figure 12B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form L prepared according to Example 12A. [Figure 12C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form L prepared according to Example 12A. [Figure 13A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form M (anhydrous) prepared according to Example 13A. [Figure 13B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form M prepared according to Example 13A. [Figure 13C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form M prepared according to Example 13A. [Figure 14A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form N (anhydrous) prepared according to Example 14A. [Figure 14B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form N prepared according to Example 14A. [Figure 14C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form N prepared according to Example 14A. [Figure 15A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form O (toluene solvate) prepared according to Example 15A. [Figure 15B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form O prepared according to Example 15A. [Figure 15C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form O prepared according to Example 15A. [Figure 16A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form P (chlorobenzene solvate) prepared according to Example 16A. [Figure 16B]1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form P prepared according to Example 16A. [Figure 16C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form P prepared according to Example 16A. [Figure 17A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form Q (1,4-dioxane solvate) prepared according to Example 17A. [Figure 17B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form Q prepared according to Example 17A. [Figure 17C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form Q prepared according to Example 17A. [Figure 18A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form R (anhydrate / hydrate) prepared according to Example 18A. [Figure 18B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form R prepared according to Example 18A. [Figure 18C] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form R prepared according to Example 18A. [Figure 19A] FIG. 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form S prepared according to Example 19A. [Figure 19B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of Compound 1 Form S prepared according to Example 19A. [Figure 20A] 2 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form T prepared according to Example 20A. [Figure 21A] FIG. 2 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 Form U (anhydrous) prepared according to Example 21A. [Figure 21B] 2 is a differential scanning calorimetry (DSC) profile of Compound 1 Form U prepared according to Example 21A. [Figure 21C]2 shows a thermogravimetric analysis (TGA) profile of Compound 1 Form U prepared according to Example 21A. [Figure 21D] 1 shows the 1H-nuclear magnetic resonance (1H-NMR) spectrum of compound 1 form U prepared according to Example 21A. [Figure 22A] 1 shows the X-ray powder diffraction (XRPD) pattern of Compound 1 amorphous form. [Figure 22B] 1 is a differential scanning calorimetry (DSC) / thermogravimetric analysis (TGA) profile of the amorphous form of Compound 1. [Figure 23] 1 illustrates the interconversion of Compound 1 crystalline forms. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0085] Unless otherwise defined, 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 belongs. All patents, patent applications and publications referenced herein are hereby incorporated by reference.

[0086] As used herein, the term "solvate" refers to a crystalline form of Compound 1 that contains a solvent.

[0087] As used herein, the terms "subject," "individual," or "patient," used interchangeably, refer to any animal, including mammals, e.g., mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and humans. In some embodiments, the patient is a human. In some embodiments, the subject is experiencing and / or exhibiting at least one symptom of a disease or disorder to be treated and / or prevented. In some embodiments, the subject is suspected of having a multi-tyrosine kinase-associated cancer.

[0088] As used herein, a "therapeutically effective amount" of a crystalline form of a salt of Compound 1 is an amount sufficient to ameliorate or in any way reduce the symptoms, or to halt or reverse the progression of a condition, or to negatively regulate or inhibit the activity of multiple tyrosine kinases. Such an amount may be administered as a single dose or according to any regimen whereby it is effective.

[0089] As used herein, the term "form" is used to describe a crystalline form and is interchangeable with the term "species". The term "crystal form" or "crystalline form" refers to a solid form that is crystalline. In certain embodiments, a crystalline form of a substance may be substantially free of amorphous forms and / or other crystalline forms. In certain embodiments, a crystalline form of a substance may contain less than about 1% by weight, less than about 2% by weight, less than about 3% by weight, less than about 4% by weight, less than about 5% by weight, less than about 6% by weight, less than about 7% by weight, less than about 8% by weight, less than about 9% by weight, less than about 10% by weight, less than about 15% by weight, less than about 20% by weight, less than about 25% by weight, less than about 30% by weight, less than about 35% by weight, less than about 40% by weight, less than about 45% by weight, or less than about 50% by weight of one or more amorphous forms and / or other crystalline forms. In certain embodiments, a crystalline form of a substance may be physically and / or chemically pure. In certain embodiments, a crystalline form of a 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 well-defined structure, such as particles lacking a crystal structure. Unless otherwise specified, the term "amorphous" or "amorphous form" means that the substance, component, or product in question is not substantially crystalline as determined by X-ray diffraction. Specifically, the term "amorphous form" describes an irregular solid form, i.e., a solid form lacking long-range crystalline order. In certain embodiments, an amorphous form of a substance is substantially free of other amorphous forms and / or crystalline forms. In certain embodiments, the amorphous form of the substance may contain, by weight, less than about 1% by weight, less than about 2% by weight, less than about 3% by weight, less than about 4% by weight, less than about 5% by weight, less than about 10% by weight, less than about 15% by weight, less than about 20% by weight, less than about 25% by weight, less than about 30% by weight, less than about 35% by weight, less than about 40% by weight, less than about 45% by weight, or less than about 50% by weight of one or more other amorphous and / or crystalline forms. In certain embodiments, the amorphous form of the substance may be physically and / or chemically pure. In certain embodiments, the amorphous 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.

[0091] As used herein, "treatment" means any manner in which the symptoms or pathology of a condition, disorder or disease are ameliorated or otherwise beneficially altered, and treatment also includes the optional use of the compositions herein as pharmaceuticals.

[0092] As used herein, amelioration of symptoms of a particular disorder by administration of a particular pharmaceutical composition refers to any reduction, whether permanent or temporary, sustained or transient, that may result from or be associated with administration of the composition.

[0093] As used herein, the term "about" when used in connection with XRPD peak positions refers to the inherent variability of the peaks depending on the instrument calibration, the process used to prepare the crystalline forms of the present invention, the age of the crystalline forms, and the type of instrument used for the analysis. The variability of the measurements used in the XRPD analysis was about ±0.1 degrees 2θ.

[0094] As used herein, the term "about" when used in relation to DSC endothermic peak appearance refers to the inherent variability of the peak depending on the instrument calibration, the method used to prepare the samples of the present invention, and the type of instrument used for the analysis. The variability of the measurements used for the DSC analysis was about ±1°C.

[0095] Common methods Unless otherwise stated, the general methods outlined below were used in the illustrated examples.

[0096] I. Crystallization techniques The crystalline forms disclosed herein may be prepared using a variety of methods known to those skilled in the art, such as crystallization or recrystallization from a suitable solvent or by sublimation. A variety of techniques may be used for crystallization or recrystallization, such as evaporation of a water-miscible or water-immiscible solvent or solvent mixture, seeding with a supersaturated solution, reducing the temperature of a solvent mixture, or freeze-drying a solvent mixture, including those techniques illustrated in the Examples.

[0097] The crystallization disclosed herein may be performed with or without crystal seeds, which may be derived from any of the aforementioned batches of the desired crystalline form, e.g., 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. [Table 1]

[0098] Instruments and parameters For XRPD analysis, a PANalytical Empyrean and X'Pert3 X-ray powder diffractometer were used to characterize the physical forms obtained in this disclosure unless otherwise indicated. The XRPD parameters used are listed below. [Table 2]

[0099] A Bruker D8 advanced X-ray powder diffractometer or equivalent was also used for XRPD analysis to characterize Form A and Form U. The XRPD parameters used are listed below. [Table 3]

[0100] TGA and DSC were used to determine the physical forms obtained in this disclosure unless otherwise specified. TGA data was collected using a Ta Q500 / Q5000 TGA manufactured by TA Instruments, and DSC was performed using a Ta Q200 / Q2000 DSC manufactured by TA Instruments. The details of the parameters used are listed below. [Table 4]

[0101] Several instruments were also used for the TGA and DGA analysis of Form A or Form U. TGA data was collected using NETZSCH TG 209 F1 Instruments, and DSC was performed using TA Q 20 or TA DSC 250 Instruments. The detailed parameters used are listed below. [Table 5]

[0102] The DVS in the form obtained in this disclosure was measured via DVS Intrinsic from SMS (Surface Measurement Systems) without any special instructions (Method A). The relative humidity at 25°C was calibrated against the deliquescence points of LiCl, Mg(NO3)2 and KCl. The parameters of the DVS test are listed below. [Table 6]

[0103] DVS of Form A and Form U were also measured via DVS Intrinsic from SMS (Surface Measurement Systems) (Method B). Relative humidity at 25° C. was calibrated against the deliquescence points of LiCl, Mg(NO3)2 and KCl. The parameters of the DVS test are listed below. [Table 7]

[0104] 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. [Table 8]

[0105] The following examples are intended to illustrate further specific embodiments of the invention, but are not intended to limit the scope of the invention. EXAMPLES

[0106] 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]nonan-7-yl)benzamide (Compound 1) are known. For example, International Publication No. WO2019 / 210828 provides a detailed synthetic route for the preparation of Compound 1.

[0107] Example 1A: Preparation of Compound 1 Form A (Form A) Compound 1 (40 g) was dissolved in DCM (120 mL). The solution was concentrated to dryness, and then EA (250 mL) was added. The resulting mixture was warmed to 60-70° C. and slowly cooled to 15-25° C., followed by filtration. The resulting cake was dried at 40-50° C. for 16 h to obtain Compound 1 Form A (about 40 g), which could be used as crystal seed.

[0108] Compound 1 (8.1 kg) was dissolved in DCM (58 kg) at 20-30° C. The solution was concentrated to approximately half the volume of the mixture, after which the solution was charged with EA (45 kg) and crystal seeds (0.035 kg) were added. After stirring at 20-30° C. for 1 h, the solution was concentrated and the EA solvent mixture was exchanged with EA three times (43 kg+43 kg+24 kg). The mixture was heated to 60-70° C. and stirred for 2 h, followed by slow cooling to 15-25° C.

[0109] The resulting mixture was introduced with MeOH (32 kg) at 45-55°C and stirred for 16 h. The solvent was exchanged with MeOH three times (20 kg+21 kg+20 kg) and then the mixture was returned to EA solution by exchanging with EA three times (23 kg+47 kg+40 kg). The mixture was warmed to 60-70°C and stirred for 2.5 h followed by slow cooling to 15-25°C. The resulting mixture was cooled slowly to 15-25°C and filtered. The resulting cake was washed with EA (9 kg) and dried at 45-55°C for 18.5 h to obtain the product as a yellow solid. The solid was sieved and then a total of 7.36 kg of compound form A was obtained.

[0110] The resulting Form A was characterized using X-ray powder diffraction (XRPD) patterns (performed on a Bruker D8 advanced X-ray powder diffractometer), which showed that Form A was a crystalline form (see FIG. 1A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 1A. [Table 9-1] [Table 9-2]

[0111] Compound Form A 1 The H NMR spectrum is shown in Figure 1D. The DSC / TGA curve (performed on a NETZSCH TG 209 F1 Instrument and TA Q 20) showed that a weight loss of 8.8% up to 160°C and two endothermic peaks at 149.6°C and 178.2°C (peaks) were observed (Figures 1B and 1C). The XRPD overlay showed that after Form A was heated to 160°C, cooled to room temperature, and re-exposed to air conditions, Form A transformed into Form B. The TGA data and 1 Combined with the 1 H NMR results, it was speculated that Form A is an EtOAc solvate.

[0112] Compound 1 Form A was stepwise isothermally analyzed in a nitrogen atmosphere by TGA. When the weight loss reached 0.02%, the system was equilibrated at a constant temperature until the weight loss was <0.002%. After stepwise heating of Form A to 100°C, the results showed that the TGA weight loss was consistent with the weight loss detected by linear heating. After cooling to room temperature, the less crystalline Form B was obtained.

[0113] The DVS cycle was performed at 25° C. (Method B), and the adsorption and desorption were allowed to vary during the complete DVS cycle. Compound 1 Form A is slightly hygroscopic, with water adsorption of 0.4% at 95% RH humidity.

[0114] Example 1B: Preparation of Compound 1 Form A Compound 1 (7.0 g) was added to EA (140 mL) and then heated to reflux for 2 h. The mixture was slowly cooled to room temperature (RT), stirred for 0.5 h, filtered, washed with EA and dried under reduced pressure to give the product (4.9 g).

[0115] Example 1C: Preparation of Single Crystals of Compound 1 Form A Compound 1 (2.8 mg) was dissolved in 0.5 mL of THF / EtOAc (1:2, v / v) solvent mixture. After slow evaporation, single crystals of Compound 1 Form A were obtained.

[0116] A single crystal of Compound 1 Form A (EtOAc solvate) was characterized by SCXRD. The calculated XRPD of the single crystal structure is in close agreement with the experimental XRPD of Form A single crystal (FIG. 1E).

[0117] The single crystal was analyzed by single crystal X-ray diffractometer. The crystal system of the single crystal is triclinic and the space group is P1. The lattice 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}.

[0118] 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 with a 1:1 molar ratio of compound 1 to EtOAc, and adjacent compound 1 molecules are bonded to each other through intermolecular hydrogen bonds.

[0119] Example 2A: Preparation of Compound 1 Form B Compound 1 (20 mg) was dissolved in acetone. The mixture was filtered, and the resulting clear solution was then slowly evaporated at room temperature to obtain Form B.

[0120] The resulting Form B was characterized using an XRPD pattern, which showed that Form B was a crystalline form (see FIG. 2A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 2A. [Table 10]

[0121] The TGA / DSC curve showed that a weight loss of 3.3% up to 110 °C and two endothermic peaks at 107.7 °C and 187.3 °C (peaks) were detected before decomposition (Figure 2B). 1 Approximately 2.2% acetone was observed in the H NMR spectrum (FIG. 2C). Form B was heated to 160° C., after which no change in form was observed.

[0122] The VT-XRPD results showed that after Form B was heated to 150 °C in a N2 atmosphere and cooled to 30 °C, no morphological change was observed, suggesting that Form B was anhydrous. 1 The acetone detected by 1 H NMR was presumably caused by solvent residues.

[0123] Moreover, after Form B was heated to 160 °C, cooled to room temperature, and then heated to 160 °C again, Form B with high crystallinity could be obtained (Figure 2D). The TGA / DSC curve showed that a weight loss of 2.8% up to 150 °C and one endothermic peak at 186.5 °C (peak) was observed before decomposition. And the signal of acetone was 1 It was not detected in the 1 H NMR spectrum.

[0124] Example 2B: Preparation of Compound 1 Form B Compound 1 Form B was obtained by any one of the following procedures: 1) Heating Form A to 160° C. followed by natural cooling to room temperature; 2) stepwise isothermally heating Form A to about 100°C; 3) Form D is heated to 130° C. and isothermal for 30 minutes; 4) Form J is heated to 130° C. and isothermal for 30 minutes; or 5) Form B is heated to 160°C, cooled to room temperature, and subsequently heated to 160°C.

[0125] Example 2C: Preparation of Compound 1 Form B Compound 1 Form K (6.0 g) was added to heptane (100 mL) followed by slurried for 24 hours at reflux at a temperature of about 100° C. The mixture was cooled to room temperature, filtered, washed with heptane and dried under reduced pressure to give the product (5.5 g).

[0126] Example 3A: Preparation of Compound 1 Form C (Form C) Compound 1 (20 mg) was dissolved in MEK. The mixture was filtered, and the resulting clear solution was then slowly evaporated at room temperature to obtain Form C.

[0127] The resulting Form C was characterized using X-ray powder diffraction (XRPD) pattern, which showed that Form C was a crystalline form (see FIG. 3A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 3A. [Table 11]

[0128] The TGA / DSC curve showed a weight loss of 8.1% up to 160° C. and two endothermic peaks at 142.5° C. and 177.3° C. (peak) (FIG. 3B). 1 The H NMR spectrum (FIG. 3C) showed that the theoretical weight of MEK was calculated to be 5.4%, which is smaller than the TGA weight loss. 1 We speculate that this was caused by solvent loss during storage prior to H NMR testing. Heating experiments were performed on Form C to determine if the weight loss was due to solvent absorption.

[0129] Comparison of XRPDs showed that after heating Form C to 160° C., cooling to room temperature, and re-exposing to air conditions, Form C converted to the weakly crystalline Form B. Form C was speculated to be a MEK solvate.

[0130] Example 4A: Preparation of Compound 1 Form D (Form D) Amorphous Compound 1 (20 mg) was suspended in IPAc and the suspension was slurried by stirring at room temperature for 1-7 days to obtain Form D.

[0131] The resulting Form D was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that Form D was a crystalline form (see FIG. 4A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 4A. [Table 12]

[0132] The TGA / DSC data showed a weight loss of 7.2% up to 130° C. and three endothermic peaks at 108.4° C., 160.1° C., and 177.3° C. (FIG. 4B). 1 H NMR spectrum showed that the theoretical IPAc content was determined to be 5.4%, suggesting that there may have been some solvent loss during storage (Figure 4C).

[0133] In the XRPD overlay of the heating experiment, a morphological change to form D was observed after heating form D to 165° C. and cooling to room temperature, therefore, it was inferred that form D is an IPAc solvate.

[0134] Furthermore, the results of the heating experiment showed that after heating Form D to 130° C. and isothermal for 30 minutes, a less crystalline Form B with additional peaks was obtained.

[0135] Example 5A: Preparation of Compound 1 Form E (Form E) Compound 1 (20 mg) was dissolved in anisole. The mixture was filtered, and the resulting clear solution was then slowly evaporated at room temperature to obtain Form E.

[0136] The resulting form E was characterized using X-ray powder diffraction (XRPD) pattern, which showed that form E was a crystalline form (see FIG. 5A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 5A. [Table 13]

[0137] The TGA curve showed a weight loss of 11.9% up to 180° C., and the DSC curve showed one endothermic peak at 157.4° C. (peak) before decomposition (FIG. 5B). 1 Based on the H NMR spectrum (Figure 5C), we determined approximately 17.1% anisole. This was higher than the weight loss from TGA and was speculated to be caused by heterogeneous solvent residues. The results of the heating experiment showed that a decrease in crystallinity was observed after heating form E to 170 °C and subsequent cooling, suggesting that the endothermic peak on the DSC curve may be a signal of melting. We speculated that form E is an anisole solvate.

[0138] Example 6A: Preparation of Compound 1 Form F Amorphous Compound 1 (20 mg) was suspended in 0.5 mL of EtOH and stirred at 50° C. to obtain Form F.

[0139] The resulting form F was characterized using X-ray powder diffraction (XRPD) pattern, which showed that form F was a crystalline form (see FIG. 6A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 6A. [Table 14-1] [Table 14-2]

[0140] The TGA / DSC curve showed a weight loss of 0.8% up to 80° C., one broad peak around 69.7° C. before decomposition, and two endothermic peaks at 156.8° C. and 177.8° C. (peaks) (FIG. 6B). 1 No EtOH signal was detected in the H NMR spectrum (FIG. 6C). The results of the heating experiment showed that no morphological change was observed when form F was heated to 80° C., and the diffraction peaks of form B were detected after heating form F to 150° C. and 165° C. According to the VT-XRPD results, no morphological change was observed after heating form F to 100° C. in N2 and cooling to 30° C., indicating that form F was anhydrous. In the DSC, the broad endotherm observed at 69.7° C. was speculated to be caused by the loss of residual solvent or moisture, and the endotherm at 156.8° C. could be related to the morphological transformation at high temperature.

[0141] Example 7A: Preparation of Compound 1 Form G (Form G) Amorphous Compound 1 (20 mg) was suspended in MTBE, and the suspension was slurried by stirring at room temperature for 1-7 days to obtain Form G.

[0142] The resulting form G was characterized using X-ray powder diffraction (XRPD) pattern, which showed that form G was a crystalline form (see FIG. 7A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 7A. [Table 15]

[0143] The TGA / DSC results showed that a weight loss of 4.6% up to 160 °C and one weak endothermic peak at 117.2 °C and one strong endothermic peak at 157.7 °C (peak) were observed before decomposition (Figure 7B). 1 Combined with the H NMR results (Figure 7C), the theoretical weight of MTBE was calculated to be 5.1%. XRPD overlays before and after heating showed that a clear decrease in crystallinity was observed after the heating experiment. Form G was assumed to be an MTBE solvate.

[0144] Example 8A: Preparation of Compound 1 Form H (Form H) Amorphous Compound 1 (20 mg) was suspended in ACN, and the suspension was slurried by stirring at room temperature for 1-7 days to obtain Form H.

[0145] The resulting Form H was characterized using X-ray powder diffraction (XRPD) pattern, which showed that Form H was a crystalline form (see FIG. 8A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 8A. [Table 16]

[0146] The TGA / DSC curve showed that a weight loss of 1.2% up to 170° C. and three endothermic peaks at 60.1° C., 162.9° C., and 179.5° C. (peaks) were detected before decomposition (FIG. 8B). 1 No ACN signal was detected in the H NMR spectrum (FIG. 8C), indicating that form H may be an anhydrate / hydrate.

[0147] Example 9A: Preparation of Compound 1 Form I (Form I) Amorphous Compound 1 (20 mg) was suspended in 0.5 mL of IPA and stirred at 50° C. to obtain Form I.

[0148] The obtained Form I was characterized using X-ray powder diffraction (XRPD) pattern, which showed that Form I was a crystalline form (see FIG. 9A). [Table 17]

[0149] The TGA / DSC curve showed that a weight loss of 2.1% up to 120° C. and two endothermic peaks at 134.0° C. and 159.7° C. before decomposition were detected (FIG. 9B). 1 In the H NMR spectrum (FIG. 9C), IPA peaks were observed and the content was calculated to be 3.2%. Comparison of XRPD showed that a clear decrease in crystallinity was observed for Form I during the heating experiment. Form I was assumed to be an IPA solvate.

[0150] Example 9B: Preparation of Compound 1 Form I Form I, slowly evaporated in acetone, showed an XRPD pattern identical to that of Form I of Example 9A. Two TGA weight losses (1.9% up to 110° C. and 2.7% from 110° C. to 200° C., see FIG. 9D) and two endothermic peaks at 78.0° C. and 160.3° C. prior to decomposition were observed in the DSC thermogram. 1 H NMR (FIG. 9E) results showed that the acetone content in the sample was determined to be 2.8%.

[0151] According to the results of the heating experiment, no morphological change was observed after heating Form I from acetone to 130 °C, but an amorphous sample was observed when the heating temperature reached 180 °C. TGA data and 1 Combined with the H NMR data, the first step TGA weight loss could be the desorption of volatile components, while the second step weight loss could be due to the loss of acetone, which led to the transformation to an amorphous phase. Therefore, we speculate that form I from acetone is an acetone solvate.

[0152] Since the different solvates had identical XRPD patterns as Form I, it was inferred that isomorphism occurred during the formation of Form I.

[0153] Example 10A: Preparation of Compound 1 Form J (Form J) Amorphous Compound 1 (20 mg) was suspended in 2-MeTHF / n-heptane (1:1, v / v). The suspension was slurried by stirring at room temperature for 1-7 days to obtain Form J.

[0154] The resulting form J was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form J was a crystalline form (see FIG. 10A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 10A. [Table 18]

[0155] The TGA / DSC curve showed that a weight loss of 8.0% up to 160° C. and two endothermic peaks at 125.3° C. and 175.2° C. (peak) were detected before decomposition (FIG. 10B). 1 H NMR (FIG. 10C) results showed that 2-MeTHF and n-heptane signals were observed in Form J (theoretical weight loss: about 10.2%). XRPD overlay showed that Form J was converted to Form B after heating to 150° C. and cooling to room temperature. TGA, 1 Based on 1 H NMR and heating experimental data, Form J was predicted to be a 2-MeTHF solvate.

[0156] Further, Form J was heated to 130° C., followed by isothermal at 130° C. for 30 minutes, and then cooled to room temperature. XRPD results showed that less crystalline Form B was obtained.

[0157] Example 11A: Preparation of Compound 1 Form K (Form K) Amorphous Compound 1 (20 mg) was suspended in methyl acetate and slurried by stirring at room temperature for 1-7 days to obtain Form K.

[0158] The resulting form K was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form K was a crystalline form (see FIG. 11A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 11A. [Table 19]

[0159] TGA / DSC showed that there was a 5.8% weight loss up to 120° C. and two endothermic peaks at 112.1° C. and 177.7° C. (peak) before decomposition (FIG. 11B). 1 In the 1 H NMR (FIG. 11C), a signal for methyl acetate was observed, and the theoretical weight loss was about 2.5%.

[0160] The XRPD overlay of the heating experiment showed that after heating form K to 120° C., weakly crystalline form B was observed. As the XRPD overlay showed, after storing form K at room temperature in a closed HPLC vial for about 5 weeks, form K converted to the less crystalline form B. Form K was speculated to be a methyl acetate solvate.

[0161] Example 11B: Preparation of Compound 1 Form K 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 room temperature 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).

[0162] Example 12A: Preparation of Compound 1 Form L (Form L) 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] The resulting form L was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form L was a crystalline form (see FIG. 12A). The characteristic peaks and the percentage of peak intensities obtained from the XRPD analysis are shown in Table 12A. [Table 20]

[0164] The TGA / DSC curves showed that a weight loss of 2.2% up to 100°C was observed in the TGA plot, and multiple signals were detected in the DSC curve, including four endothermic peaks at 53.7°C, 62.7°C, 76.3°C, and 162.1°C (peaks) and one exothermic peak at 89.6°C before decomposition (Figure 12B). 1 Based on the H NMR spectrum (FIG. 12C), no acetone peaks were observed, therefore, form L was likely anhydrous / hydrate.

[0165] Form L converted to another form upon storage at room temperature, and Form L converted to Form I. Therefore, Form L was speculated to be a metastable anhydrate / hydrate that can be desolvated from the wet cake from the solvent system.

[0166] Example 13A: Preparation of Compound 1 Form M (Form M) Amorphous Compound 1 (20 mg) was suspended in CHCl3 / n-heptane (1:1, v / v). The suspension was exposed to a temperature cycle from 50°C to 5°C to obtain Form M.

[0167] The resulting Form M was characterized using X-ray powder diffraction (XRPD) pattern, which showed that Form M was a crystalline form (see FIG. 13A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 13A. [Table 21]

[0168] The TGA / DSC curve showed a weight loss of 1.6% up to 170° C. and one endothermic peak at 171.0° C. (peak) before decomposition (FIG. 13B). 1Based on the H NMR results, no obvious signal of CHCl3 was observed (Figure 13C). The VT-XRPD results showed that after Form M was heated to 120 °C in N2 and cooled to 30 °C, no morphological change was observed, suggesting that Form M was anhydrous.

[0169] Example 14A: Preparation of Compound 1 Form N (Form N) Amorphous Compound 1 (20 mg) was suspended in 0.5 mL of ACN and stirred at 50° C. to obtain Form N.

[0170] The resulting form N was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form N was a crystalline form (see FIG. 14A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 14A. [Table 22]

[0171] The TGA / DSC curve showed that a weight loss of 0.3% up to 160° C. and one endothermic peak at 160.6° C. (peak) was observed before decomposition (FIG. 14B). 1 The H NMR results showed that there was no ACN signal (Figure 14C). 1 Combining the 1 H NMR data, it was inferred that Form N is anhydrous.

[0172] Example 15A: Preparation of Compound 1 Form O (Form O) 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] The resulting Form O was characterized using an X-ray powder diffraction (XRPD) pattern, which indicated that Form O was a crystalline form (see FIG. 15A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 15A. [Table 23-1] [Table 23-2]

[0174] The TGA / DSC curve showed that a weight loss of 8.9% up to 160° C. and four endothermic peaks at 115.8° C., 117.7° C., 146.6° C., and 175.8° C. (peaks) were observed before decomposition (FIG. 15B). 1 In the H NMR spectrum (Figure 15C), toluene peaks were observed and the theoretical weight loss was determined to be 11.0%. The high theoretical weight loss may have been caused by inhomogeneous solvent residues. The results of the heating experiment showed that after heating Form O to 160 °C and cooling to room temperature, Form O transformed into Form B. The TGA data and 1 Combined with the H NMR data, the species O was inferred to be a toluene solvate.

[0175] Example 16A: Preparation of Compound 1 Form P (Form P) Compound 1 (20 mg) was dissolved in chlorobenzene and centrifuged. The supernatant was exposed to toluene at room temperature to obtain Form P.

[0176] The resulting form P was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form P was a crystalline form (see FIG. 16A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 16A. [Table 24]

[0177] TGA / DSC results showed a 9.9% weight loss up to 140° C. and one endothermic peak at 121.6° C. (peak) before decomposition (FIG. 16B). 1 Combined with the H NMR results (Figure 16C), the theoretical weight of chlorobenzene was calculated to be 9.8%, which was consistent with the weight loss from the TGA. Comparison of the XRPD showed that some of the diffraction peaks had disappeared after about 4 weeks of storage at room temperature. After the sample was heated to 140 °C, even more diffraction peaks had disappeared. TGA,1 Combining 1 H NMR data and heating experiments, Form P is predicted to be a chlorobenzene solvate.

[0178] Example 17A: Preparation of Compound 1 Form Q (Form Q) Amorphous Compound 1 (20 mg) was subjected to solid vapor diffusion in 1,4-dioxane at room temperature for 10 days to give Form Q.

[0179] The resulting form Q was characterized using an X-ray powder diffraction (XRPD) pattern, which indicated that form Q was a crystalline form (see FIG. 17A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 17A. [Table 25]

[0180] TGA / DSC showed a 9.0% weight loss up to 160° C. and one endothermic peak at 155.1° C. (peak) before decomposition (FIG. 17B). 1 In the H NMR spectrum (FIG. 17C), a peak of 1,4-dioxane was detected, with a theoretical weight of 5.6%. The theoretical weight loss was smaller than the TGA weight loss, which may have been caused by solvent loss during storage. Form Q was predicted to be a 1,4-dioxane solvate.

[0181] Example 18A: Preparation of Compound 1 Form R (Form R) Amorphous Compound 1 (approximately 100 mg) was suspended in 0.5 mL of ACN to obtain Form R.

[0182] The resulting form R was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that form R was a crystalline form (see FIG. 18A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 18A. [Table 26]

[0183] The TGA / DSC curve showed a weight loss of 2.8% up to 120° C., 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. (FIG. 18B). 1 No ACN signal was observed in the H NMR spectrum (FIG. 18C). VT-XRPD showed that no morphological change was observed after Form R was dried with N2 for about 20 minutes, and additional peaks and obvious peak shifts were observed after Form R was heated to 100° C. in N2 and cooled to 30° C. Considering the complex heat signal observed before 100° C. in DSC, Form R was assumed to be anhydrous / hydrate.

[0184] Example 19A: Preparation of Compound 1 Form S (Form S) Compound 1 form R was heated to 150° C. in a N2 atmosphere and subsequently cooled to 30° C. to obtain form S.

[0185] The resulting Form S was characterized using an X-ray powder diffraction (XRPD) pattern, which indicated that Form S was a crystalline form (see FIG. 19A). The characteristic peaks and percentages of peak intensities obtained from the XRPD analysis are shown in Table 19A. [Table 27-1] [Table 27-2]

[0186] The TGA / DSC curve showed that a weight loss of 1.7% up to 120° C. and two endothermic peaks at 93.8° C. and 169.5° C. (peak) were observed before decomposition (FIG. 19B).

[0187] Example 20A: Preparation of Compound 1 Form T (Form T) After Form N was kept under 25° C. / 60% RH and 40° C. / 75% RH for 1 week and sealed at 80° C. for 24 hours, Form N converted to Form T. However, after 3 days of storage under the same conditions, Form T converted back to Form N.

[0188] The resulting Form T was characterized using an X-ray powder diffraction (XRPD) pattern, which showed that Form T was a crystalline form (see FIG. 20A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 20A. [Table 28]

[0189] XRPD overlay of the transformation indicated that Form T could be anhydrous / hydrate.

[0190] Example 21A: Preparation of Compound 1 Form U (Form U) To a solution of Compound 1 (40 g) in DCM (240 mL) at 20-40° C., n-heptane (140 mL) was slowly added. Stirred for 1 h, then another batch of n-heptane (20 mL) was added followed by holding for 0.5 h. Subsequently, n-heptane (20 mL) was added and held for 0.5 h, followed by n-heptane (20 mL). Finally, n-heptane (40 mL) was added and stirred at 20-40° C. for 12 h. The mixture was filtered and the resulting cake was dried at 45-55° C. for 18 h to obtain Compound 1 Form U (36.5 g), which could be used as a crystal seed.

[0191] To a solution of Compound 1 (6.6 kg) in DCM (51 kg) at 25-35° C., n-heptane (14 kg) was added followed by crystal seeds (0.020 kg). The mixture was stirred at 20-35° 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 followed by stirring at 20-35° C. for about 2 hours each. The mixture was then stirred at 20-35° C. for about 16 hours. The mixture was filtered and washed with n-heptane (13 kg), followed by drying the resulting cake at 45-55° C. for 30 hours to obtain Compound 1 Form U (5.86 kg) as a yellow solid.

[0192] The resulting form U was characterized using X-ray powder diffraction (XRPD) patterns (performed on a Bruker D8 advanced X-ray powder diffractometer), which showed that form U was a crystalline form (see FIG. 21A). The characteristic peaks and peak intensity percentages obtained from the XRPD analysis are shown in Table 21A. [Table 29-1] [Table 29-2]

[0193] As shown by the TGA / DSC curve (performed on a NETZSCH TG 209 F1 Instrument, TA DSC 250), a weight loss of 0.2% up to 150° C. and one endothermic peak at 170.8° C. (peak) were detected (FIGS. 21B and 21C). 1 No DCM signals were detected in the 1 H NMR spectrum (Figure 21D).

[0194] DVS cycling was performed at 25° C. (Method B), and sorption and desorption were allowed to vary during a complete DVS cycle. Compound 1 form U was slightly hygroscopic, with water sorption of 1.4% at 95% RH humidity.

[0195] As in the synthesis of compound 1 shown in International Patent Publication WO2019 / 210828, the acid intermediate (S)-2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(2-(2-(2-isopropylphenyl)pyrrolidin-1-yl)-7-azaspiro[3.5]nonan-7-yl)benzoic acid was reacted with the sulfamide intermediate 4-((((1r,4r)-4-hydroxy-4-methylcyclohexyl)methyl)amino)-3-nitrobenzenesulfonamide to obtain compound 1. At the same time, due to the azaindole moiety of compound 1 reacting with the acid intermediate, a dimeric compound as a process impurity could be generated. In the production, unexpectedly, only in form U can the dimeric impurity be effectively removed. In one production batch, the content of the dimeric impurity in the in-process control (IPC) was 0.4% (by weight). After working up by EA crystallization, form A was obtained, and the content of dimer impurity was still 0.4%. After further working up by recrystallization from a THF / ACN mixture solution, the content of dimer impurity was reduced to 0.22%. Finally, after working up by recrystallization from a DCM / heptane mixture solution, form U was obtained, and no further dimer impurity was detected.

[0196] Example 21B: Preparation of Compound 1 Form U Amorphous Compound 1 (20 mg) was suspended in a mixture of DCM / n-heptane (1:1, v / v) at room temperature. The suspension was slurried by stirring at room temperature for 1-7 days to obtain Form U.

[0197] Example 21C: Preparation of Compound 1 Form U 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 heptane (5.0 mL) at 40° C. The mixture was cooled to room temperature and stirred to produce a precipitate. The precipitate was filtered, washed with heptane and dried to give the product (1.4 g).

[0198] Example 22A: Compound 1 Amorphous Form (Amorphous) 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 slurried with MTBE (1.0 L) and filtered. The filter cake was collected and dried under vacuum to obtain the product (137.5 g).

[0199] The resulting amorphous form exhibited the X-ray powder diffraction (XRPD) pattern in Figure 22A. TGA / DSC results (Figure 22B) showed that two stages of weight loss (0.7% up to 110°C, 0.5% from 110°C to 200°C) and a possible glass transition signal at 126.7°C (center) were observed. The chemical purity was determined to be 98.3% by high performance liquid chromatography (HPLC). DVS results showed it to have a water uptake of 1.8% at 80% RH / 25°C.

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

[0201] XRPD overlays showed that no morphological changes between forms B, S, and M were observed.

[0202] No morphological changes were observed for Form R after 1 week of storage at 25°C / 60% RH or 40°C / 75% RH. After 24 hours of storage at 80°C / closed, Form R transformed to a morphology similar to Form S.

[0203] For Form F, as shown by the XRPD overlays, a mixture of Form F and Form B was observed after storage of the Form F samples at all conditions tested.

[0204] XRPD pattern overlays showed that no morphological change was observed for Form H after storage under 25 °C / 60% RH or 40 °C / 75% RH for 1 week, but an obvious decrease in crystallinity was observed after storage under 80 °C / closed for 24 h.

[0205] Form N converts to form T, which can convert back to form N upon storage at room temperature for about 3 days.

[0206] Solubility of Solid Forms The solubility of different physical forms of Compound 1 was tested in water, 0.1 N HCl, acetate buffer at pH 4.5, and phosphate buffer at pH 6.8. At 24 hours, the concentration of Compound 1 was detected by HPLC.

[0207] For Compound 1 amorphous, Compound 1 was not detected in water, pH 4.5 buffer, and pH 6.8 buffer, while the corresponding solubility in 0.1 N HCl was 35.30 μg / ml. For Compound 1 Form A, 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, while in 0.1 N HCl it was 29.36 μg / ml. Thus, Form A showed higher solubility in water, pH 4.5 buffer, and 6.8 buffer, but lower solubility in 0.1 N HCl, when compared to the amorphous form.

[0208] solid form stability The solid state stability of Form B was evaluated in an acetone / HO system at 50° C. Approximately 2 mg of Form B sample was used to slurry or shake in acetone / HO (1:9, v / v) and HO solution (saturated with amorphous sample).

[0209] The crystalline state under slurry was investigated and the XRPD overlay showed: 1) After Form B was slurried in acetone / H2O (1:9, v / v) or H2O for about 4 days, a decrease in the crystallinity of Form B, including amorphous content, was observed. 2) After Form B was slurried in H2O for approximately 4 hours, no morphology change was observed.

[0210] Furthermore, the crystalline state under shaking was also investigated. XRPD overlays showed that no morphological changes were observed after shaking Form B in acetone / HO (1:9, v / v) or HO for about 4 hours or 4 days, suggesting that Form B could be affected by mechanical force.

[0211] Physical and chemical stability tests Long-term and accelerated stability studies were performed on different physical forms of Compound 1 by storing samples at 25±2°C / 60±5%RH and 40±2°C / 75±5%RH conditions for up to 6 months, and each sample was tested by HPLC for total impurity content.

[0212] For the amorphous Compound 1, the chemical purity of Compound 1 was significantly decreased, for example, the total impurity content increased from 2.1% to 4.2% when stored at 40±2°C / 75±5%RH for 6 months, and many new impurities were detected.

[0213] For Compound 1 Form A, the chemical purity of Compound 1 did not have a significant change. For example, when stored at 40±2°C / 75±5% RH for 6 months, the total content of impurities only increased from 0.40% to 0.52%. Furthermore, no change in crystal form or optical purity was observed, but the content of the solvent EA increased from about 9.5 to 8.8 (x10 4 ppm).

[0214] For Compound 1 Form U, the chemical purity of Compound 1 had no significant change. For example, the total content of impurities only increased from 0.40% to 0.72% upon storage for 6 months at 40±2°C / 75±5%RH conditions. Furthermore, no changes in crystal form and optical purity were observed.

[0215] Thus, both Form A and Form U of Compound 1 exhibited better physical stability and Form A exhibited better chemical stability compared to the amorphous form of Compound 1.

[0216] While the invention has been described in relation to specific embodiments thereof, it is understood that further modifications are possible, and this application is generally intended to cover any variations, uses, or adaptations of the invention in accordance with the principles of the invention, and includes such departures from the present disclosure as are known or within customary practice in the art to which the invention pertains and may be applied to the essential features described above, as well as those departures from the present disclosure as fall within the scope of the appended claims.

Claims

1. 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]nonan-7-yl)benzamide (Compound 1) which is an EtOAc solvate containing about 1 mole of EtOAc per mole, said crystalline form designated as Form A.

2. i) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 16.5±0.1° and 24.5±0.1°; or ii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 12.4±0.1°, 16.5±0.1°, and 24.5±0.1°; or iii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 12.4±0.1°, 16.5±0.1°, 20.7±0.1°, and 24.5±0.1°; or iv) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 10.6±0.1°, 12.4±0.1°, 16.5±0.1°, 20.7±0.1°, and 24.5±0.1°; or v) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or vi) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or vii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or viii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or ix) The crystalline form of claim 1, wherein the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°.

3. 3. The crystalline form of any one of claims 1-2, wherein Form A has an XRPD pattern substantially as shown in Figure 1A or Figure 1E.

4. 3. The crystalline form of any one of claims 1 to 2, wherein Form A is characterized by having two endothermic peaks at about 150°C and about 178°C by differential scanning calorimetry (DSC).

5. 3. The crystalline form of any one of claims 1-2, wherein Form A has a DSC thermogram substantially as shown in Figure 1B.

6. 3. The crystalline form of any one of claims 1-2, wherein the crystalline system of Form A is triclinic and the space group is P1 with lattice parameters of (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)°.

7. A crystalline form of anhydrous 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]nonan-7-yl)benzamide (Compound 1), said crystalline form designated as Form B.

8. i) the crystalline form has an X-ray powder diffraction pattern comprising a diffraction peak having a °2θ angle value at 14.4±0.1°; or ii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1° and 17.5±0.1°; or iii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1°, 17.5±0.1°, and 18.4±0.1°; or iv) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 14.4±0.1°, 17.5±0.1°, 18.4±0.1°, and 19.6±0.1°; or v) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of °2θ values ​​at 7.2±0.1°, 14.4±0.1°, 17.5±0.1°, 18.4±0.1°, and 19.6±0.1°; or vi) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or vii) The crystalline form of claim 7, wherein the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°.

9. 9. The crystalline form of any one of claims 7-8, wherein Form B has an XRPD pattern substantially as shown in Figure 2A or Figure 2D.

10. 9. The crystalline form of any one of claims 7 to 8, wherein Form B is characterized by having two endothermic peaks at about 187°C by differential scanning calorimetry (DSC).

11. 9. The crystalline form of any one of claims 7-8, wherein Form B has a DSC thermogram substantially as shown in Figure 2B.

12. A crystalline form of anhydrous 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]nonan-7-yl)benzamide, said crystalline form designated as Form U.

13. i) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 11.3±0.1° and 24.3±0.1°; or ii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 11.3±0.1°, 15.6±0.1°, and 24.3±0.1°; or iii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 11.3±0.1°, 15.6±0.1°, 21.2±0.1°, and 24.3±0.1°; or iv) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 11.3±0.1°, 13.5±0.1°, 15.6±0.1°, 21.2±0.1°, and 24.3±0.1°; or v) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or vi) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or vii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or viii) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or ix) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or x) the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°; or xi) The crystalline form of claim 12, wherein the crystalline form has an X-ray powder diffraction pattern comprising diffraction peaks having °2θ angle values ​​at 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°.

14. 14. The crystalline form of any one of claims 12-13, wherein form U has an XRPD pattern substantially as shown in Figure 21A.

15. 14. The crystalline form of any one of claims 12-13, wherein form U is characterized by having one endothermic peak at about 171°C by differential scanning calorimetry (DSC).

16. 14. The crystalline form of any one of claims 12-13, wherein Form U has a DSC thermogram substantially as shown in Figure 21B.

17. 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]nonan-7-yl)benzamide (Compound 1).

18. 18. A pharmaceutical composition comprising: (a) a therapeutically effective amount of a crystalline form of any one of claims 1, 2, 7, 8, 12, and 13, or an amorphous form of Compound 1 of claim 17; and (b) one or more pharmaceutically acceptable excipients.

19. A composition comprising the crystalline form of any one of claims 1, 2, 7, 8, 12, and 13 or the amorphous form of compound 1 of claim 17; or (a) 18. A pharmaceutical composition comprising the crystalline form of any one of claims 1, 2, 7, 8, 12 and 13, or the amorphous form of compound 1 of claim 17, and (b) one or more pharmaceutically acceptable excipients, wherein the composition or pharmaceutical composition is for use in treating a disease associated with Bcl-2 protein inhibition.

20. Do one of the following: a) Dissolving Compound 1 in DCM, removing DCM, and charging with EA to obtain Form A; b) Compound 1 is dissolved in DCM, concentrated, charged with EA, and the DCM is exchanged with EA, MeOH, and EA separately to obtain Form A; c) dissolving Compound 1 in EA, heating and cooling to obtain Form A; or 3. The crystalline form of any one of claims 1 to 2, obtained by a process comprising: d) dissolving Compound 1 in a THF / EtOAc (1:2, v / v) solvent mixture and evaporating to obtain Form A.

21. Do one of the following: a) dissolving Compound 1 in acetone and evaporating the solvent to obtain Form B; b) heating Form A, Form C, and Form O to about 160° C. and cooling to obtain Form B; c) stepwise isothermal heating of Form A to about 100°C to obtain Form B; d) heating Form D or Form J to about 130° C. and isothermal to obtain Form B; or e) adding Form K in heptane, heating to about 100° C. and cooling to obtain Form B.

22. Do one of the following: a) dissolving Compound 1 in DCM and adding n-heptane to the batch and stirring to obtain Form U; or 14. The crystalline form of any one of claims 12 to 13, obtained by a process comprising: b) dissolving compound 1 in a mixture of DCM / n-heptane (1:1, v / v) and stirring to obtain form U.

23. Do one of the following: a) Compound 1 is dissolved in DCM and dried, or 18. The amorphous form of claim 17, obtained by a process comprising: b) dissolving Compound 1 in a mixture of solvents containing DCM and drying to obtain said amorphous form.

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