Crystalline forms of 3 - (imidazo [1, 2-b] pyridazin-3-ylethynyl) - 4-methyl-n - {4 - [(4-methylpiperazin-1-yl) methyl] - 3 - (trifluoromethyl) phenyl} benzamide and its monohydrochloride

Abstract

To provide polymorphs of 3 - (imidazo [1, 2-b] pyridazin-3-ylethynyl) - 4-methyl-N - {4 - [(4-methylpiperazin-1-yl) methyl] - 3 - (trifluoromethyl) phenyl} benzamide, which is a multi-targeted tyrosine kinase inhibitor useful for the treatment of diseases including chronic myelogenous leukemia (CML).SOLUTION: Novel crystalline 3 - (imidazo [1, 2-b] pyridazin-3-ylethynyl) - 4-methyl-N - {4 - [(4-methylpiperazin-1-yl) methyl] - 3 - (trifluoromethyl) phenyl} benzamide monohydrochloride in Forms A to K.SELECTED DRAWING: Figure 12

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 61 / 736,543, filed December 12, 2012; U.S. Provisional Patent Application No. 61 / 737,007, filed December 13, 2012; and U.S. Provisional Patent Application No. 61 / 788,208, filed March 15, 2013, which are incorporated herein by reference in their entireties. [Background technology]

[0002] The present application relates to novel crystalline forms of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide and 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride, compositions containing such crystalline forms, and methods for their preparation and use. 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide has the chemical formula C, corresponding to a formula weight of 532.56 g / mol. 29 H 27 It has the chemical structure F3N6O, shown below: [ka]

[0003] The CAS registry number for 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide is 943319-70-8.

[0004] 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride has the chemical formula C, which corresponds to a formula weight of 569.02 g / mol. 29 H 28 It has the chemical structure ClF3N6O, shown below: [ka]

[0005] The CAS registry number for 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride is 1114544-31-8.

[0006] The United States Adopted Name (USAN) and International Nonproprietary Name (INN) for 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide is ponatinib. Alternative chemical names for ponatinib include benzamide, 3-(2-imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-[4-[(4-methyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl], and 3-[2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl]-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide.

[0007] The United States Adopted Name (USAN) and International Nonproprietary Name (INN) for 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride is ponatinib hydrochloride. Alternative chemical names for ponatinib hydrochloride include benzamide, 3-(2-imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-[4-[(4-methyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-, hydrochloride (1:1), and 3-[2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl]-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride.

[0008] Ponatinib is a multitargeted tyrosine kinase inhibitor useful in the treatment of diseases including chronic myeloid leukemia (CML). Ponatinib hydrochloride is a small molecule pan-BCR-ABL inhibitor in clinical development for the treatment of adult patients with chronic-phase, accelerated-phase, or blast phase CML or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) who are resistant or intolerant to prior tyrosine kinase inhibitor therapy. Other tyrosine kinase inhibitors relevant to the treatment of such CML or Ph+ALL include GLEEVEC® (imatinib mesylate) and TASIGNA® (nilotinib) (both Novartis AG), SPRYCEL® (dasatinib) (Bristol Myers Squibb), and BOSULIF® (bosutinib) (Pfizer). A New Drug Application (NDA) for ponatinib hydrochloride was submitted to the U.S. FDA on July 30, 2012. The U.S. FDA approved the NDA on December 14, 2012, and ponatinib hydrochloride is now commercially available under the brand name ICLUSIG® (ponatinib).

[0009] In addition, ponatinib hydrochloride may be clinically useful in treating other disorders or conditions involving the inhibition of other protein kinases. Such kinases and resulting disorders or conditions are described in O'Hare, T., et al., Cancer Cell, Vol. 16, No. 5, 401-412 (2009) and WO 2011 / 053938, which are incorporated herein by reference for all purposes.

[0010] Understanding the possible polymorphic forms of active pharmaceutical ingredients, such as ponatinib and ponatinib hydrochloride, is useful in drug development. Not knowing the specific polymorphic forms present or desired in an API can lead to inconsistent manufacturing of the API, potentially resulting in variations in drug performance across different lots of the API. Furthermore, for many of the same reasons, it is important to discover as many polymorphic forms of an API as possible so that their stability can be systematically characterized over time. Once a specific polymorphic form has been selected for drug development, it is important to be able to reproducibly prepare that polymorphic form. Additionally, because impurities can affect drug performance, it is desirable to have a process for producing APIs, such as ponatinib and ponatinib hydrochloride, with high purity.

[0011] The earliest patent publication known to the applicant to disclose the chemical structure of ponatinib hydrochloride is International Publication No. WO 2007 / 075869, also owned by the applicant (ARIAD Pharmaceuticals), which is incorporated herein by reference for all purposes. Example 16 of WO 2007 / 075869 describes the product being obtained as a solid with m / z (M+H) of 533, which corresponds to the mass of the free base of ponatinib. Example 16 also describes the preparation of the monohydrochloride salt of ponatinib. There is no specific mention in Example 16 that the obtained ponatinib hydrochloride was crystalline, nor is there any specific crystalline form of ponatinib hydrochloride specified.

[0012] U.S. Patent Application No. 11 / 644,849, published as U.S. Patent Application Publication No. 2007 / 0191376, is a counterpart of WO 2007 / 075869, granted February 14, 2012, as U.S. Patent No. 8,114,874, which is incorporated herein by reference for all purposes. U.S. Patent Application Publication No. 13 / 357,745 is a continuation of U.S. Patent Application No. 11 / 644,849, also incorporated herein by reference for all purposes.

[0013] Other patent applications published as of the filing date of the present application that address ponatinib hydrochloride include WO 2011 / 053938 and WO 2012 / 139027, which are incorporated herein by reference for all purposes. Like WO 2007 / 075869, WO 2011 / 053938 and WO 2012 / 139027 do not specify a particular crystalline form of ponatinib hydrochloride. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] International Publication No. 2007 / 075869 Summary of the Invention

[0015] It has now been determined that both ponatinib and ponatinib hydrochloride can exist in polymorphic forms, including certain crystalline forms, some of which are suitable for pharmaceutical formulation development.

[0016] In one aspect, the present disclosure relates to polymorphs of ponatinib, which are designated herein as Forms A, B, C, D, E, F, G, H, I, J, and K.

[0017] In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a polymorph of ponatinib disclosed herein and at least one pharmaceutically acceptable carrier, excipient, or vehicle.

[0018] In yet another aspect, the present disclosure relates to substantially pure crystalline forms of ponatinib hydrochloride, which are designated herein as Forms A, B, C, D, E, F, G, H, I, J, and K.

[0019] In yet another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of a substantially pure crystalline form of ponatinib hydrochloride disclosed herein and at least one pharmaceutically acceptable carrier, excipient, or vehicle.

[0020] In yet another aspect, the disclosure provides a process for preparing a substantially pure crystalline form of ponatinib hydrochloride by contacting ponatinib with hydrochloric acid.

[0021] In another aspect, the disclosure relates to a method of treating a human disorder or condition that responds to the inhibition of a protein kinase by administering to the human a therapeutically effective amount of a polymorph of ponatinib disclosed herein, hi certain embodiments, the disorder or condition is chronic myeloid leukemia (CML).

[0022] In yet another aspect, the disclosure relates to a method of treating a human disorder or condition that responds to the inhibition of a protein kinase, by administering to the human a therapeutically effective amount of a substantially pure crystalline form of ponatinib hydrochloride disclosed herein. In certain embodiments, when the protein kinase is Bcr-Abl or a mutant thereof, the disorder or condition is chronic myeloid leukemia (CML) or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL).

[0023] The following drawings form part of the present specification and are included to demonstrate certain aspects of the present invention in detail. The invention may be better understood by reference to one or more of these drawings and the detailed description of specific embodiments presented herein. [Brief explanation of the drawings]

[0024] [Figure 1] 1 is a summary of the 11 solid state forms of ponatinib hydrochloride, including HCl polymorphs and pseudopolymorphs, that have been discovered and identified as Forms A through K and are disclosed herein. [Figure 2]

[0023] Figure 2 is a summary of the specific solid forms of ponatinib hydrochloride discovered and identified in Figure 1 and disclosed herein. The legend for Figure 2 is as follows: a Starting material: Form HCl or amorphous material (Am) obtained by lyophilization. b Occ: Total occurrence includes 216 experiments performed in Phase 2 (described in Example 1 herein), in which 39 samples were further analyzed wet or the mother liquor was evaporated and analyzed, resulting in a total of 254 materials characterized. For example, "(3, 1.2%)" corresponds to the form appearing in 3 of 254 measurements, representing a percentage of 1.2%. For 62 of 254 measurements (9%), the product yield was too low to identify a solid form or the material was wet. d Am: Amorphous form. [Figure 3] Characteristic X-ray powder diffraction (XRPD) patterns of two batches of ponatinib hydrochloride Form A, each obtained before and after DVS humidity cycling, with relative intensity (expressed in counts) on the vertical axis and angle (2θ) on the horizontal axis. [Figure 4] 1 is a characteristic X-ray powder diffraction (XRPD) pattern obtained from ponatinib hydrochloride Form A, with relative intensity (expressed in counts) on the vertical axis and angle (2θ) on the horizontal axis. [Figure 5] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained from ponatinib hydrochloride Form A. The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 6]1 is a characteristic thermogravimetric analysis (TGA) and thermogravimetric analysis coupled with mass spectrometry (TGMS) scan of volatiles obtained from Form A of ponatinib hydrochloride. [Figure 7] 1 is a characteristic H-NMR spectrum (600 MHz) of ponatinib hydrochloride in solution obtained from Form A of ponatinib hydrochloride in DMSO-d6 at 300 K. The vertical axis shows normalized intensity, and the horizontal axis shows chemical shifts (ppm). [Figure 8] 1 shows a characteristic F-NMR spectrum (564 MHz) of ponatinib hydrochloride in solution obtained from Form A of ponatinib hydrochloride in DMSO-d6 at 300 K. The vertical axis shows normalized intensity, and the horizontal axis shows chemical shifts (ppm). [Figure 9] 1 is a characteristic C-NMR spectrum (151 MHz) of ponatinib hydrochloride in solution obtained from Form A of ponatinib hydrochloride in DMSO-d6 at 300 K. The vertical axis shows normalized intensity, and the horizontal axis shows chemical shifts (ppm). [Figure 10] Characteristic mass spectral patterns obtained from Form A of ponatinib hydrochloride. The upper mass spectral pattern is the observed mass of Form A, and the lower mass spectral pattern is the spectrum of the daughter ion of the parent shown in the upper Form A. The vertical axis shows relative abundance, and the horizontal axis shows atomic mass (m / z). [Figure 11] 1 is a characteristic mass spectral fragmentation pattern of ponatinib hydrochloride form A. The vertical axis shows relative abundance and the horizontal axis shows atomic mass (m / z). [Figure 12] 12 depicts the structure of Form A of ponatinib hydrochloride according to the data set forth in the table herein entitled "Crystalline Data and Structure Refinement for Form A of Ponatinib Hydrochloride." Atoms in this Figure 12 are color-coded according to atomic type: carbon gray, nitrogen blue, oxygen red, hydrogen white, fluorine yellow, and chlorine green. [Figure 13] 1 is a characteristic FT-IR spectrum obtained from ponatinib hydrochloride Form A. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm). [Figure 14]1 is a characteristic HPLC spectrum obtained from ponatinib hydrochloride Form A. The vertical axis shows absorbance units (mAU) and the horizontal axis shows time (minutes). [Figure 15] FIG. 1 shows a characteristic X-ray powder diffraction (XRPD) pattern obtained from ponatinib hydrochloride Form A (bottom) compared with the XRPD patterns of Forms B (middle) and C (top), with relative intensity (expressed in counts) on the vertical axis and angle (2θ) on the horizontal axis. [Figure 16] 1 is a characteristic HPLC spectrum obtained from ponatinib hydrochloride form B. The vertical axis shows absorbance units (mAU) and the horizontal axis shows time (minutes). [Figure 17] Characteristic X-ray powder diffraction (XRPD) patterns obtained from ponatinib hydrochloride Form C (top) compared to the XRPD pattern for Form A (bottom). Relative intensity (expressed as counts) is plotted on the vertical axis, and angle (2θ) is plotted on the horizontal axis. [Figure 18] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained from ponatinib hydrochloride Form C. The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 19] 1 is a characteristic thermogravimetric analysis (TGA) scan obtained from Form C of ponatinib hydrochloride. [Figure 20] A characteristic TGMS thermogram obtained from Form C of ponatinib hydrochloride. [Figure 21] 1 is a characteristic HPLC spectrum obtained from ponatinib hydrochloride Form C. The vertical axis shows absorbance units (mAU) and the horizontal axis shows time (minutes). [Figure 22] 1 shows a characteristic X-ray powder diffraction (XRPD) pattern obtained from Form D of ponatinib hydrochloride compared to the XRPD patterns of Form A and certain other crystalline forms within the HCl3 class. The vertical axis shows relative intensity (expressed in counts) and the horizontal axis shows angle (2θ). [Figure 23] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained from Form D of ponatinib hydrochloride, with heat flow (mW) on the vertical axis and temperature (°C) on the horizontal axis. [Figure 24]1 is a characteristic thermogravimetric analysis (TGA) scan obtained from Form D of ponatinib hydrochloride. [Figure 25] A characteristic FT-IR spectrum obtained from Form D of ponatinib hydrochloride. The vertical axis shows percent transmittance (%), while the horizontal axis shows wavenumber (cm-1). Form A starting material is shown in red, and Form D (PSM1) is shown in blue. [Figure 26] A characteristic HPLC spectrum obtained from Form D of ponatinib hydrochloride, with absorbance units (mAU) on the vertical axis and time (minutes) on the horizontal axis. [Figure 27] 1 is a characteristic X-ray powder diffraction (XRPD) pattern obtained from Form F of ponatinib hydrochloride compared to the XRPD patterns of Form A and certain other crystalline forms within the HCl5 class. The vertical axis shows relative intensity (expressed in counts) and the horizontal axis shows angle (2θ). [Figure 28] FIG. 1 shows two characteristic differential scanning calorimetry (DSC) scans obtained from Form F of ponatinib hydrochloride. The top scan is the DSC curve for VDS1. The bottom scan is the DSC curve for VDS2. The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 29] Characteristic thermogravimetric analysis and overlay of SDTA (top) and TGMS (bottom) scans obtained from Form F of ponatinib hydrochloride (VDS1). [Figure 30] Overlay of characteristic thermogravimetric (top) and TGMS (bottom) scans obtained from Form F of ponatinib hydrochloride (VDS2). [Figure 31] A characteristic FT-IR spectrum obtained from Form F of ponatinib hydrochloride. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm-1). Form A starting material is shown in red, and Form F (VDS1) is shown in green. [Figure 32] A characteristic FT-IR spectrum obtained from Form F of ponatinib hydrochloride. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm-1). Form A starting material is shown in purple, and Form F (VDS2) is shown in red. [Figure 33]A characteristic HPLC spectrum obtained from Form F of ponatinib hydrochloride (VDS2) with absorbance units (mAU) on the vertical axis and time (min) on the horizontal axis. [Figure 34] A characteristic X-ray powder diffraction (XRPD) pattern obtained from Form H of ponatinib hydrochloride compared to the XRPD patterns of Form A (bottom) and certain other crystalline forms within the HCl6 class. Relative intensity (expressed in counts) is plotted on the vertical axis and angle (2θ) is plotted on the horizontal axis. [Figure 35] Characteristic differential scanning calorimetry (DSC) scans obtained from Form H of ponatinib hydrochloride. The top scan is the DSC curve for VDS3. The bottom scan is the DSC curve for VDS4. The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 36] Overlay of characteristic thermogravimetric (top) and TGMS (bottom) scans obtained from Form H of ponatinib hydrochloride (VDS3). [Figure 37] Overlay of characteristic thermogravimetric (top) and TGMS (bottom) scans obtained from Form H of ponatinib hydrochloride (VDS4). [Figure 38] A characteristic FT-IR spectrum obtained from Form H of ponatinib hydrochloride. The vertical axis shows percent transmittance (%), while the horizontal axis shows wavenumber (cm-1). Form A starting material is shown in purple, and Form H (VDS3) is shown in red. [Figure 39] A characteristic FT-IR spectrum obtained from Form H of ponatinib hydrochloride. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm-1). Form A starting material is shown in purple, and Form H (VDS4) is shown in red. [Figure 40] A characteristic HPLC spectrum obtained from Form H of ponatinib hydrochloride (VDS4) with absorbance units (mAU) on the vertical axis and time (minutes) on the horizontal axis. [Figure 41]Figure 1 shows an overlay of characteristic X-ray powder diffraction (XRPD) patterns for each solid form identified in Figure 1, where the vertical axis represents relative intensity (counts) and the horizontal axis represents 2-theta (degrees). The solid forms and solvents in this figure, from bottom to top, are as follows: starting material ponatinib HCl (HCl1) (Form A), Form HCl2 (QSA12.1, solvent: water) (Form B), Form HCl2b (QSA21.1, solvent: water) (Form C), Form HCl3-class (GRP12.1, solvent: toluene) (Form D), mixture HCl1 + HCl4 (GRP1.1, solvent: hexafluorobenzene) (Form E), Form HCl5 (VDS28.1, solvent: butyl acetate) (Form F), Form HCl5b (after drying VDS28.2, solvent: butyl acetate) (Form G), and Form HCl6-class (VDS6.1, solvent: methanol) (Form H). [Figure 42] Figure 1 shows representative digital images of (top to bottom, left to right): HCl 6 class (VDS6.1, vds050.0c:E1), HCl form 5 (VDS28.1, vds05.0c:B3), HCl form 5b (VDS28.2, vds05.1c:B6), mixture HCl 1 + HCl 4 (GRP1.1, grp02.0c:A1), HCl 3 class form (GRP12.1, grp02.1c:L1), HCl form 2 (QSA12.1, qsa00.1c:A2), and HCl form 2b (QSA21.1, qsa00.1c:J2). [Figure 43] A characteristic X-ray powder diffraction (XRPD) pattern obtained from Form J of ponatinib hydrochloride, with relative intensity (expressed in counts) on the vertical axis and angle (2θ) on the horizontal axis. [Figure 44] A characteristic X-ray powder diffraction (XRPD) pattern obtained from Form K of ponatinib hydrochloride, with relative intensity (expressed in counts) on the vertical axis and angle (2θ) on the horizontal axis. [Figure 45] FIG. 1 shows characteristic XRPD patterns of Form A of ponatinib hydrochloride (lower pattern) and amorphous ponatinib hydrochloride (upper pattern) (solvent: 2,2,2-trifluroethanol), where the vertical axis represents relative intensity (counts) and the horizontal axis represents 2-theta (degrees). [Figure 46]This is a characteristic differential scanning calorimetry (DSC) thermogram of amorphous 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride. A strong endothermic event was observed with a peak at 259.4 °C, corresponding to the melting point of this amorphous form. The vertical axis represents heat flow (mW) and the horizontal axis represents temperature (°C). [Figure 47] A characteristic HPLC spectrum obtained from Form H of ponatinib hydrochloride (VDS4). The vertical axis shows absorbance units (mAU) and the horizontal axis shows time (min). The vertical axis shows absorbance units (mAU) and the horizontal axis shows time (min). [Figure 48] 1 is a table summarizing the solid-state forms of ponatinib, including polymorphs and pseudopolymorphs identified as Forms A through J. [Figure 49] 1 is a table summarizing the occurrence rate, crystallization method, physical form, endotherm, and purity of polymorphs and pseudopolymorphs of ponatinib identified as Forms A to J. [Figure 50] FIG. 1 shows the molecular structure and numbering scheme of Form A of ponatinib free base crystalline (anhydrate) as determined by single crystal X-ray analysis. [Figure 51] FIG. 1 shows simulated and experimentally obtained XRPD patterns of Form A of ponatinib free base crystalline (anhydrous). [Figure 52] Characteristic differential scanning calorimetry (DSC) scan obtained from crystalline Form A of ponatinib free base (anhydrous), with heat flow (mW) on the vertical axis and temperature (°C) on the horizontal axis. [Figure 53] FIG. 1 shows the molecular structure and numbering scheme of the B-class ponatinib / 1,4-dioxane (1:1) solvate (Form B) as determined by single crystal X-ray analysis. [Figure 54] FIG. 1 is a simulated XRPD pattern of the B-class ponatinib / 1,4-dioxane (1:1) solvate (Form B). [Figure 55]FIG. 1 shows the molecular structure and numbering scheme of the B-class ponatinib / perfluorobenzene (1:1) solvate (Form B) as determined by single crystal X-ray analysis. [Figure 56] B-Class: Simulated and experimentally obtained XRPD patterns of ponatinib / perfluorobenzene (1:1) solvate (Form B). [Figure 57] FIG. 1 shows the XRPD patterns of Form B of ponatinib / 2-methylTHF (1:0.4) solvate (top pattern); Form B of ponatinib / cyclohexanone (1:1) solvate (middle pattern); and Form A (bottom pattern). [Figure 58] Characteristic differential scanning calorimetry (DSC) scan obtained from Class B 1:1 ponatinib / cyclohexanone solvate (QSA7.1). Heat flow [mW] is shown on the vertical axis and temperature (°C) is shown on the horizontal axis. [Figure 59] Characteristic TGA and SDTA thermogram overlay of B-Class 1:1 ponatinib / cyclohexanone solvate (QSA7.1). [Figure 60] Characteristic differential scanning calorimetry (DSC) scan obtained from B Class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1). The vertical axis shows heat flow [mW] and the horizontal axis shows temperature (°C). [Figure 61] 1 is an overlay of characteristic TGA and SDTA thermograms of B Class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1). [Figure 62] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained from poorly crystalline Form C of ponatinib (GEN3.1). The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 63] 1 is a characteristic TGA and SDTA thermogram overlay of poorly crystalline form C of ponatinib (GEN3.1). [Figure 64] FIG. 1 shows the XRPD patterns of poorly crystalline Form C of ponatinib (GEN3.1) (top pattern) and Form A (bottom pattern). [Figure 65]Characteristic differential scanning calorimetry (DSC) scan obtained from Form D ponatinib (GEN5.1R1). The vertical axis shows heat flow (mW) and the horizontal axis shows temperature (°C). [Figure 66] 1 is an overlay of characteristic TGA and SDTA thermograms of d-form ponatinib (GEN5.1R1). [Figure 67] Figure 1 shows the XRPD patterns of Form D ponatinib (middle pattern) and Form A (bottom pattern), as well as the XRPD patterns of a B-class ponatinib solvate and Form D ponatinib mixture (top pattern). [Figure 68] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained from E-clasponatinib / THF 1:1 solvate (GEN7.1), with heat flow [mW] on the vertical axis and temperature (°C) on the horizontal axis. [Figure 69] 1 is an overlay of characteristic TGA and SDTA thermograms of E-clasponatinib / THF 1:1 solvate (GEN7.1). [Figure 70] FIG. 1 shows the XRPD patterns of E-classponatinib / THF 1:1 solvate (GEN7.1) (top pattern); E-classponatinib / chloroform solvate (SLP3.1) (middle pattern); and Form A (bottom pattern). [Figure 71] 1 is an overlay of characteristic TGA and SDTA thermograms of form F ponatinib (AS16.2). [Figure 72] Characteristic differential scanning calorimetry (DSC) scan obtained from Form H (VLD1, dried solid from stock). Heat flow [mW] is shown on the vertical axis and temperature (°C) is shown on the horizontal axis. [Figure 73] Overlay of characteristic TGA and SDTA thermograms of Form H (VLD1, dry solid from stock). [Figure 74]Figure 2 shows a series of XRPD patterns: Plot 1 is Form H (VLD2 experiment, after 2 weeks and drying); Plot 2 is Form H (VLD1 experiment, after 2 weeks and drying); Plot 3 is Form H (VLD1 experiment, dried solid from stock after DVS); Plot 4 is Form H (VLD1, dried solid from stock); Plot 5 is Form H (VLD19); The bottom plot is the XRPD of Form A. [Figure 75] Figure 1 shows an overlay of the characteristic FT-IR spectrum obtained from ponatinib Form H with the FT-IR spectrum obtained from Form A. The vertical axis shows percent transmittance (%), and the horizontal axis shows wavenumber (cm). [Figure 76] This figure shows an overlay of the characteristic FT-IR spectrum from 1750 to 600 nm obtained from ponatinib Form H and Form A. The vertical axis shows percent transmittance (%), and the horizontal axis shows wavenumber (cm-1). [Figure 77] A characteristic differential scanning calorimetry (DSC) scan obtained from Form I (GEN9.1) shows heat flow (mW) on the vertical axis and temperature (°C) on the horizontal axis. [Figure 78] 1 is an overlay of characteristic TGA and SDTA thermograms of Form I (GEN9.1). [Figure 79] FIG. 1 shows the overlaid XRPD patterns of ponatinib Form I (top pattern) and Form A (bottom pattern). [Figure 80] Figure 1 shows an overlay of the following XRPD patterns: Plot 6 is the XRPD pattern of Form A (VDS2, after stability testing); Plot 7 is the XRPD pattern of Form J (VDS2); Plot 8 is the XRPD pattern of Form J (VDS10 of Screening S10010A); Plot 9 is another XRPD pattern of Form A of ponatinib. [Figure 81] FIG. 1 shows a characteristic FT-IR spectrum obtained from Form J overlaid with the FT-IR spectrum obtained from Form A. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm). [Figure 82]This figure shows the characteristic FT-IR spectrum obtained from Form J superimposed over the wavelength region of 1750 to 600 nm on the FT-IR spectrum obtained from Form A. The vertical axis shows percent transmittance (%), and the horizontal axis shows wavenumber (cm-1). [Figure 83] Figure 1 shows an overlay of XRPD results obtained for selected ponatinib free base polymorphs (from bottom to top): Form A (SM); B-class (QSAS7.1); Form C, poorly crystalline (GEN3.1); Form D (GEN5.1); E-class (SLP3.1); Form F (SLP10.1); Form G (AS19.1); Form H (VDL19.1); Form I, poorly crystalline (GEN9.1); and Form J, poorly crystalline (VDS10.1). [Figure 84] FIG. 1 shows an overlay of XRPD patterns of the free base starting material, with the two patterns representing two different batches (F09-05575: lower pattern; and F09-05576: upper pattern). [Figure 85] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained for ponatinib free base starting material batch F09-05575, with heat flow [mW] on the vertical axis and temperature (° C.) on the horizontal axis. [Figure 86] 1 is a characteristic differential scanning calorimetry (DSC) scan obtained for ponatinib free base starting material batch F09-05576, with heat flow [mW] on the vertical axis and temperature (° C.) on the horizontal axis. [Figure 87] 1 is an overlay of characteristic TGA and SDTA thermograms of ponatinib free base starting material batch F09-05575. [Figure 88] 1 is an overlay of characteristic TGA and SDTA thermograms of ponatinib free base starting material batch F09-05576. [Figure 89] 1 is a table summarizing the physical stability of several solid forms of ponatinib. [Figure 90] 1 is a table summarizing the results of scale-up experiments for selected free base forms. [Figure 91] 1 is a table summarizing various characterizations of the ponatinib free base form replicated at a 120 mg scale. DETAILED DESCRIPTION OF THE INVENTION

[0025] It has been found that both 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide and 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride can be obtained in various solid crystalline forms.

[0026] The terms "crystalline form," "polymorph," or "polymorph" may be used interchangeably herein and refer to a crystalline form of ponatinib or ponatinib hydrochloride that differs from the amorphous form of ponatinib or ponatinib hydrochloride or other form(s) of ponatinib or ponatinib hydrochloride as determined by certain physical properties such as thermodynamic stability, physical parameters, X-ray structure, DSC, and preparation process.

[0027] While polymorphism classically refers to the ability of a compound to crystallize into two or more different crystalline species (same chemical structure but distinct physicochemical properties), the term "pseudopolymorphism" is usually applied to solvate and hydrate crystalline forms. However, for the purposes of this disclosure, both true and pseudopolymorphs (i.e., hydrates and solvates) are included within the terms "crystalline form" and "polymorph." Additionally, "amorphous form" refers to an irregular solid.

[0028] It should be noted that while different samples of a particular crystalline form may have the same major XRPD peak, differences in the XRPD pattern may be observed for minor peaks. With respect to XRPD, the term "about," when used in reference to an XRPD maximum (expressed in degrees 2-theta), generally means within 0.3 degrees 2-theta of a given value. Alternatively, the term "about" can mean a value that one of ordinary skill in the art would consider to fall within an acceptable range of standard error (in this and all contexts). As used herein, the terms "isolated" and "substantially pure" mean that greater than about 50% of the crystalline ponatinib or ponatinib hydrochloride is present in the identified crystalline form (as can be determined by methods in the art) relative to the sum of other solid form(s) present in a selected substance.

[0029] Definitions and Abbreviations Solvent abbreviations: DCM: dichloromethane DMA: N,N-dimethylacetamide DMF: N,N-dimethylformamide DMSO: Dimethyl sulfoxide TFE: 2,2,2-trifluoroethanol THF: Tetrahydrofuran 2-Methyl THF: 2-methyltetrahydrofuran EtOH: Ethanol MeOH: Methanol Other abbreviations (alphabetical): ·Am: Amorphous API: Active Pharmaceutical Ingredient AS: poor solvent CI: Counterion DSC: Differential scanning calorimetry DVS: Dynamic Vapor Sorption GRP: Grinding experiment ID HPLC: High-Performance Liquid Chromatography ·HT-XRPD: High-throughput powder X-ray diffraction HR-XRPD: High-resolution powder X-ray diffraction ·LC: Low crystalline substance ·MS: Mass spectrometry PSM: ID of cooling / evaporation crystallization experiments ·SAS: Solubility evaluation SDTA: Single Differential Thermal Analysis S: Solvent SM: Starting material ·TGA: Thermogravimetric analysis TGMS: Thermogravimetric analysis combined with mass spectrometry VDL: ID for vapor diffusion experiments in liquids VDS: Vapor diffusion on solids XRPD: X-ray powder diffraction

[0030] Analysis method Powder X-ray diffraction XRPD patterns were obtained using an Avantium T2 high-throughput XRPD suite. Plates were mounted on a Bruker GADDS diffractometer equipped with a Hi-Star area detector. The XRPD platform was calibrated using silver behenate for long d-spacings and corundum for short d-spacings.

[0031] The most distinctive feature of the XRPD pattern is the 2θ region of 1.5° to 41.5°, where monochromatic CuK α Data collection was performed at room temperature using a 90-second exposure time. Diffraction patterns were collected for each well over two 2θ ranges (1.5°≦2θ≦21.5° for the first frame and 19.5°≦2θ≦41.5° for the second frame) with an exposure time of 90 seconds for each frame. XRPD patterns were not subjected to background subtraction or curve smoothing. The support material used during XRPD analysis was transparent to X-rays and produced negligible background.

[0032] thermal analysis Melting characteristics were determined from DSC thermograms recorded on a heat flux DSC822e instrument (Mettler-Toledo GmbH, Switzerland). Temperature and enthalpy of the DSC822e were calibrated using small pieces of indium (mp = 156.6 °C; ΔHf = 28.45 J / g). Samples were sealed in 40 μl standard aluminum pans, punctured, and heated in the DSC from 25 °C to 300 °C at a heating rate of 10 °C / min. Dry N gas was used to purge the DSC instrument during the measurements at a flow rate of 50 ml / min.

[0033] Mass loss due to solvent or water loss from various crystalline samples was measured by TGA / SDTA. Sample weights were monitored during heating using a TGA / SDTA851e instrument (Mettler-Toledo GmbH, Switzerland) to obtain weight versus temperature curves. The TGA / SDTA851e was temperature calibrated using indium and aluminum. Samples were weighed into 100 μl aluminum crucibles and sealed. A small hole was punctured in the seal, and the crucibles were heated in the TGA from 25 to 300 °C at a heating rate of 10 °C / min. Dry N2 gas was used for purging.

[0034] The gases released from the TGA samples were analyzed with a mass spectrometer, Omnistar GSD 301 T2 (Pfeiffer Vacuum GmbH, Germany), a quadrupole mass spectrometer that resolves masses in the range of 0–200 amu.

[0035] Digital Imaging Digital images of all wells of each well plate were collected automatically using a Philips PCVC 840K CCD camera controlled by Avantium Photoslider software.

[0036] press The compression tests were performed using an Atlas Power Press T25 (Specac), an electric hydraulic press with a maximum operating capacity of 25 tons.

[0037] HPLC analysis method HPLC analysis was performed using an Agilent 1200SL HPLC system equipped with UV and MS detectors according to the conditions given below: [Table 1]

[0038] The integrity of the compound is expressed as a peak area percentage calculated from the area of ​​each peak other than the "injection peak" in the chromatogram and the total peak area as follows: Peak area % = (peak area / total area) * 100%

[0039] The peak area percentage of the compound of interest is used as an indicator of the purity of the component in the sample.

[0040] I. Polymorphs of Ponatinib Monohydrochloride A total of 11 polymorphs of ponatinib hydrochloride have been discovered through XRPD analysis. These 11 novel polymorphs are referred to herein as HCl1 (also referred to herein as "Form A"), HCl2 (also referred to herein as "Form B"), HCl2b (also referred to herein as "Form C"), HCl3-class (also referred to herein as "Form D"), HCl1+HCl4 mixture (also referred to herein as "Form E"), HCl5-class or simply HCl5 (also referred to herein as "Form F"), HCl5b or HCl5 desolvate (also referred to herein as "Form G"), HCl6-class (also referred to herein as "Form H"), HCl6 desolvate (also referred to herein as "Form I"), HCl7 (also referred to herein as "Form J"), and HCl8 (also referred to herein as "Form K"). The nature or origin of these 11 polymorphs is shown in Figure 1. Additionally, certain characteristics of the referenced polymorphs are shown in Figure 2. For example, Form A has been shown to be an anhydrous form of ponatinib hydrochloride and was obtained as a single crystal.

[0041] In general, crystalline forms of ponatinib hydrochloride have advantageous physical properties (e.g., enhanced stability) for solid dosage commercial formulations compared to amorphous ponatinib hydrochloride. The differences between crystalline ponatinib hydrochloride and amorphous ponatinib hydrochloride are readily apparent from the same types of physicochemical data (e.g., DSC, XRPD, thermal analysis) used to distinguish the individual crystalline forms of ponatinib hydrochloride disclosed herein.

[0042] Next, with reference to the above methodology, attention is directed to each of the discovered polymorphs of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride.

[0043] Characteristics of Form A (HCl1): Anhydrous HCl-1 (the same crystalline form as the starting material) was the predominant crystalline form discovered. The chemical structure of ponatinib hydrochloride has been unambiguously confirmed by nuclear magnetic resonance spectroscopy (NMR), mass spectrometry (MS), and single crystal X-ray crystallography, combined with confirmatory data from elemental and chloride analysis, Fourier transform infrared (FT-IR) spectroscopy, and ultraviolet (UV) spectroscopy. The preferred solid form of ponatinib hydrochloride is the anhydrous crystalline HCl-1 solid form, i.e., Form A.

[0044] With reference to Figure 3, samples of ponatinib HCl, ASI batch 110020 and CGAM batch F08-06057, were analyzed by powder X-ray diffraction (XRPD). In each case, the material was analyzed before and after DVS humidity cycling. XRPD patterns were obtained using a high-throughput XRPD diffractometer. Data collection was performed at room temperature using monochromated CuKα radiation in the 2θ region from 1.5° to 41.5°, which is the most characteristic portion of the XRPD pattern. Diffraction patterns for each well were collected over two 2θ ranges (1.5° ≤ 2θ ≤ 21.5° for the first frame, and 19.5° ≤ 2θ ≤ 41.5° for the second frame) with a 90-second exposure time for each frame. No background subtraction or curve smoothing was performed on the XRPD patterns. Figure 3 shows the powder X-ray diffraction patterns for each of these materials in the HCl-1 solid form. The powder patterns are consistent with those simulated from single-crystal X-ray diffraction experiments of the HCl-1 form. XRPD data obtained before and after the DVS humidity cycling experiment demonstrate that the solid form of HCl-1 is maintained after humidity cycling. The XRPD pattern of Form A, shown in Figure 3, exhibits at least one or all of the following peaks in degrees two-theta (2θ): 5.9; 7.1; 10.0; 12.5; 16.4; 19.3; 21.8; 23.8; and 26.1. In certain embodiments, the XRPD pattern of Form A exhibits two peaks, three peaks, four peaks, or five peaks. The term "about" applies to each peak listed for this form and all other forms referred to in this disclosure.

[0045] Figure 4 shows a characteristic X-ray powder diffraction (XRPD) pattern for Form A of ponatinib hydrochloride, which appears in more detail than the XRPD. The XRPD pattern for Form A shown in Figure 4 shows at least one or more of the following peaks at angles two-theta (2θ): 5.9; 7.1; 10.0; 12.5; 13.6; 14.1; 15.0; 16.4; 17.7; 18.6; 19.3; 20.4; 21.8; 22.3; 23.8; 24.9; 26.1; 27.0; 28.4; 30.3; 31.7; and 35.1. In certain embodiments, Form A is characterized by an XRPD pattern including one or more of the following peaks at angles two-theta (2θ): 12.5; 19.3; 23.8; and 26.1. In certain embodiments, the XRPD pattern of Form A exhibits two peaks, three peaks, four peaks, or five peaks.

[0046] Differential vapor sorption (DVS) experiments using HCl-1 were performed at a constant temperature of 25°C, cycling from 45% to 95% relative humidity (sorption), 0% relative humidity (desorption), and back to 45% relative humidity with a 60-minute hold time per step. Results of this DVS experiment, performed on ponatinib HCl CGAM batch F08-060507, showed a water uptake rate of 1.1% at 95% relative humidity, while ponatinib HCl ASI batch 110020 showed a water uptake rate of 1.4% at 95% relative humidity. This water uptake rate was reversible when cycling to lower humidity levels. These results indicate that HCl-1 is not a hygroscopic compound. Additionally, the effect of humidity cycling on HCl-1 was assessed by powder X-ray diffraction (XRPD) analysis before and after the DVS experiment. XRPD data revealed that humidity cycling had no effect on the solid form of this material, and the material maintained its HCl-1 solid form.

[0047] With reference to Figure 5, the melting point of ponatinib HCl in the HCl-1 solid form was determined by differential scanning calorimetry (DSC). A sample of ponatinib HCl, ASI batch 110020, was analyzed in a crucible with a small hole, using a dry N gas purge, at a heating rate of 10°C / min over the temperature range of 25°C to 300°C. A strong endothermic event was observed, peaking at 264.1°C, corresponding to the melting point of Form A.

[0048] Referring to Figure 6, thermogravimetric analysis (TGA) and thermogravimetric analysis coupled with mass spectrometry (TGMS) of volatiles were performed on ponatinib HCl, ASI batch 110020. Samples placed in a crucible with a small hole were heated in the TGA instrument from 25 °C to 300 °C at a heating rate of 10 °C / min using dry N2 gas for purging. The gases released from the TGA were analyzed using a quadrupole mass spectrometer. The HCl-1 solid form of ponatinib HCl, ASI batch 110020, contained 0.31 wt% water and 0.85 wt% ethanol upon release. The TGA / TGMS experiments showed that mass losses of 0.2% (water) and 0.6% (ethanol from the crystallization solvent) were observed over the temperature ranges of 25 to 130 °C and 130 to 240 °C, respectively. This mass loss is consistent with the water and ethanol content upon release. Ethanol is released from the material at higher temperatures than water, but is not attributed to ponatinib HCl in the HCl-1 solid form as a solvate.

[0049] A thorough liquid-phase NMR study of Form A of ponatinib HCl was performed using a combination of multiple 1D and 2D NMR techniques, 1 H, 19 F and 13 The C resonances were fully assigned, thereby confirming the chemical structure of ponatinib HCl. Analysis of a sample of ponatinib HCl (ASI batch 110020) dissolved in deuterated DMSO (DMSO-d6) solvent was performed at ARIAD Pharmaceuticals (Cambridge, MA). NMR spectra were acquired at 300 K on a Bruker Avance III-600 MHz NMR spectrometer equipped with a 5 mm BBFO z-gradient probe. 1 The H chemical shifts were referenced to the DMSO peak at 2.5 ppm. Figure 7 shows the 1D spectra of Form A of ponatinib HCl in DMSO-d6. 1 The H-NMR spectrum is shown. 1 The H resonance 32a arises from the protonated piperazine moiety of ponatinib HCl. 1 H spectrum (Figure 7) and 13The EtOH resonances seen in both the C spectrum (Figure 9) and the C spectrum (Figure 9) arise from residual EtOH present in ponatinib HCl. Figure 8 shows the 1D structure of Form A of ponatinib HCl in DMSO-d6. 19 Figure 9 shows the 1D F-NMR spectrum of Form A of ponatinib HCl in DMSO-d6, with a characteristic chemical shift at 57.94 ppm. 13 C-NMR spectrum is shown.

[0050] 1 H and 13 Key chemical shift data for Form A of ponatinib monohydrochloride obtained from C-NMR experiments are summarized in Table 1. The number and relative intensities (integrals) of signals confirm the number of protons and carbons in the structure of Form A of ponatinib HCl. These chemical shift data are reported according to the atom numbering scheme shown immediately below. [ka]

[0051] [Table 2]

[0052] With reference to Figure 10, mass spectral experiments and collisionally activated MS2 fragmentation of Form A of ponatinib HCl were performed using a Thermo Finnegan Exactive accurate mass spectrometer and an LTQ XL ion trap mass spectrometer, respectively, operated in positive ion electrospray mode. A sample of Form A of ponatinib HCl (ASI batch 110020) dissolved in acetonitrile was introduced into the mass spectrometer via syringe pump infusion. The Exactive mass spectrometer was used in full scan mode to obtain the exact mass of ponatinib HCl. The observed mass for this infusion experiment was m / z 533.2269 (MH+), with a calculated exact mass of 533.2271 (MH+), a mass difference of 0.2 mmu (Δppm of -0.38) (Figure 10, top). The fragmentation spectrum of ponatinib HCl from an Exactive mass spectrometer is shown in Figure 10, which includes product ions from m / z 533.2269 (the molecular ion of ponatinib HCl) and ions from all other co-eluting compounds.

[0053] Figure 11 shows MS fragmentation data obtained on an LTQ XL ion trap mass spectrometer. Figure 11(A) shows a full scan MS of m / z 533 (MH+) of the injected sample. Figure 11(B) shows a full scan MS of m / z 533 (MH+) of the injected sample. 2 Scan 11 shows the fragment spectrum of the selected mass m / z 533. Figures 11(C) and 11(D) show the product ions from m / z 433 and 260, respectively, which are themselves the first product ions (molecular ions) from m / z 533.

[0054] The crystalline structure of Form A of ponatinib hydrochloride was determined using single-crystal X-ray diffraction analysis. Single crystals of the anhydrous HCl-1 form of ponatinib HCl were obtained using ponatinib HCl CGAM batch F08-06057 in a vapor diffusion crystallization process. Single crystals obtained using methanol as the solvent with ethyl acetate as the antisolvent were analyzed by single-crystal X-ray diffraction. Previous experiments have shown that this form diffracts well, leading to the elucidation of the structure of ponatinib HCl shown in Figure 12, the crystallographic parameters of which are summarized in Table 2. The terminal nitrogen of the piperazine is the protonation site of ponatinib HCl, consistent with previously described NMR analysis of ponatinib HCl. A chloride counterion is present immediately adjacent to the protonation site in the crystal structure. Based on this structural analysis, Form A was determined to be the anhydrous form.

[0055] [Table 3]

[0056] The attenuated total reflectance (ATR) FT-IR spectrum of Form A of ponatinib HCl, ASI batch 110020, is shown in Figure 13. The IR band assignments of selected ponatinib HCl based on the FT-IR shown in Figure 13 are summarized in Table 3.

[0057] [Table 4]

[0058] The FT-IR spectrum shows a functional group region spanning 4000–1300 cm-1. In the 3300–2800 cm-1 region (region A), several overlapping bands are observed, likely arising from stretching vibrations between hydrogen atoms and some other atoms, possibly amide NH stretching, aromatic CH stretching (from the imidazo-pyridazine heterocycle and phenyl groups), and aliphatic CH stretching (in the methyl and methylene groups), all present in the structure of ponatinib HCl. A weak band in the 2100–2260 cm-1 region (region B) is due to triple C–C bond stretching. A moderately intense band in the 1640–1690 cm-1 range is expected due to the amide C=O stretching vibration (amide 1), which is likely the band observed at 1669.8 cm-1 (region C). Two strong bands are observed in the 1500–1560 cm-1 range, which is due to secondary amide NH bending vibrations (amide 2) (region D). Weak to moderate bands observed in the 1300–1600 cm-1 range are due to (hetero)aromatic resonance-stabilized double C–C and double C–N bonds (ring stretching vibrations) and C–H bending vibrations (from methyl and methylene groups) (region E). Multiple bands, including a strong band at 1314.9 cm-1, are observed in the 1250–1335 cm-1 and 1250–1020 cm-1 ranges, which are expected to represent aromatic and aliphatic amine C–N stretching vibrations, respectively (regions F and G). The fingerprint region from 1300–910 cm-1 is complexed with a strong, broad band at 1122.6 cm-1 (region H), which is likely due to C–F stretching vibrations. The aromatic region, 910–650 cm-1, is primarily due to out-of-plane bending vibrations of the heteroaromatic C–H bonds, indicating the heteroaromatic nature of the compound (Region I). The FT-IR spectral data presented here support the proposed structure of Form A of ponatinib hydrochloride.

[0059] Experiments were performed to determine the purity of Form A. With reference to Figure 14, the purity of Form A of ponatinib hydrochloride was determined to be 99.8160% (area percent).

[0060] Characteristics of B-form (HCl2): Solubility evaluation in TFE / water yielded form HCl2, which was confirmed by repeat XRPD measurements of selected samples, converting to form HCl2b after 1 day of storage of the measurement plates under ambient conditions. HCl2 was also obtained from aqueous solvent systems (water and MeOH / water) in experiments performed in the second phase described herein, which also converted to form HCl2b during storage under ambient conditions (see overview in Figure 2).

[0061] Form B was analyzed by powder X-ray diffraction (XRPD). Figure 15 shows (from bottom to top): the XRPD patterns of the starting material (Form A), Form HCl2 (Form B) (QSA12.1, solvent: water), and Form HCl2b (Form C) (QSA12.2, re-measured after several days under ambient conditions). In the XRPD pattern shown in Figure 15, Form B exhibits at least one or all of the following peaks at angles two-theta (2θ): 3.1; 6.5; 12.4; 13.8; 15.4; 16.2; 17.4; 18.0; 20.4; 23.2; 24.4; 26.1; and 26.9. For reference, in the XRPD pattern shown in Figure 15, Form C exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.5; 12.4; 13.8; 17.4; 18.0; 20.6; 22.0; 23.0; 25.5; 26.5; and 27.4. In certain embodiments, Form B is characterized by an XRPD pattern that includes one or more of the following peaks 2-theta (2θ): 13.8; 15.4; 17.4; 18.0; 26.1; and 26.9. In such embodiments, the XRPD patterns of Forms B and C exhibit two peaks, three peaks, four peaks, or five peaks.

[0062] Experiments were performed to determine the purity of Form B. With reference to Figure 15, the purity of Form B of ponatinib hydrochloride was determined to be 99.7535% (area percent).

[0063] Characteristics of Form C (HCl2b): Form C is a hydrated form. Form HCl2b was initially obtained by conversion of Form B from solubility studies over many days at ambient conditions or directly from TFE / water solvent mixtures. Form C was also obtained from aqueous solvent systems (water and water / DMSO) in second-stage experiments (see overview in Figure 2).

[0064] Form C was analyzed by powder X-ray diffraction (XRPD). Figure 17 shows (from bottom to top): the XRPD patterns of the starting material (HCl1) and Form HCl2b (QSA21.1, solvent: water). In the XRPD pattern shown in Figure 17, Form C exhibits at least one or all of the following peaks at angles 2-theta (2θ): 3.1; 6.5; 12.4; 13.8; 17.4; 18.0; 20.6; 22.0; 23.0; 25.5; 26.5; 27.4; 28.4; and 29.0. In certain embodiments, Form C is characterized by an XRPD pattern including one or more of the following peaks 2-theta (2θ): 13.8; 17.4; 18.0; and 25.5. In certain embodiments, the XRPD pattern of Form C exhibits two peaks, three peaks, four peaks, or five peaks.

[0065] With reference to Figure 18, the melting point of Form C of ponatinib HCl was determined by differential scanning calorimetry (DSC). Samples were analyzed in small-hole crucibles with a dry N gas purge at a heating rate of 10°C / min over the temperature range of 25°C to 300°C. peak = 122.9℃, T peak = 158.2°C and T peak A strong endothermic event was observed at =256.2°C.

[0066] Figure 19 shows the TGA and SDTGA thermograms of QSA21.1. Figure 20 shows the TGMS thermogram of Form C of Experiment QSA21.1. A mass loss (water) of 4.3% is observed in the temperature interval 40°C to 140°C. The API:water ratio was estimated to be 1:1.4.

[0067] Experiments were performed to determine the purity of Form C. With reference to Figure 21, the purity of Form C of ponatinib hydrochloride was determined to be 99.7850% (area percent).

[0068] Characteristics of D-type (HCl3 class): The HCl3 class was mostly obtained from aromatic solvents, with the exception of MeOH / acetonitrile mixtures, as outlined in Figure 2. Form D was successfully reproduced on a 120 mg scale using the cooling-vapor crystallization method in toluene.

[0069] Based on thermal analysis, a representative sample of Form D was designated the toluene-solvated form (AP:toluene 1:0.5). This form desolvated and recrystallized at 199.5 °C, and a second melting was observed at 257.6 °C (approximately corresponding to the melting point of Form A). The HCl3 class is slightly hygroscopic, with a water mass uptake of 2.5% at 95% RH. This process was reversible with respect to physical stability and sample appearance.

[0070] The HCl3 class samples were found to be physically stable after 8 months of storage under ambient conditions and after DVS cycling. However, in a humidity chamber (40°C / 75% RH), the HCl3 class samples converted to HCl1 after 1 week.

[0071] Form D was analyzed by powder X-ray diffraction (XRPD). Figure 22 shows an overlay of the XRPD patterns for (bottom to top): HCl1 (AP24534 HCl salt starting material), HCl3 class (PSM17, solvent: toluene), HCl3 (PSM1, solvent: toluene), HCl1 + HCl3 (PSM1 after 1 week at 40°C / 75% RH), and HCl3 (PSM1 after DVS). In the XRPD pattern shown in Figure 22, HCl3 exhibits at least one or all of the following two-theta (2θ) peaks: 8.2; 10.1; 10.9; 14.9; 16.0; 16.3; 16.8; 17.7; 18.7; 20.2; 22.9; 24.0; 25.6; 26.7; and 28.5. In certain embodiments, Form D is characterized by an XRPD pattern including one or more of the following peaks at angles 2-theta (2θ): 8.2; 10.1; 14.9; and 25.6. In the XRPD pattern shown in Figure 22, HCl3 + HCl1 exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.5; 7.4; 12.5; 13.6; 14.1; 16.7; 17.4; 18.0; 19.3; 20.4; 21.8; 24.0; 25.1; 26.3; and 28.0. In certain embodiments, HCl3 + HCl1 is characterized by an XRPD pattern including one or more of the following peaks at angles 2-theta (2θ): 12.5; 19.3; and 26.3. In certain embodiments, the XRPD pattern of Form D exhibits two peaks, three peaks, four peaks, or five peaks.

[0072] With reference to Figure 23, the melting point of Form D ponatinib HCl (PSM1) was determined by differential scanning calorimetry (DSC). Samples were analyzed in small-hole crucibles with a dry N2 gas purge at a heating rate of 10°C / min over the temperature range of 25°C to 300°C. peak = 199.5℃, T peak = 204.1°C and T peak A strong endothermic event was observed at =257.6°C.

[0073] 24, the TGA and SDTGA thermograms of Form D (PSM1) are shown. A mass loss of 7.7% (toluene, API:solvent ratio 1:0.51) was observed in the temperature interval 120°C to 220°C.

[0074] Regarding Figure 25, 1750~500cm -1 The FT-IR spectrum of Form D is shown in the region of Figure 1. These data support the proposed structure of Form D of ponatinib hydrochloride. Furthermore, the spectrum shows unique features of Form D compared to Form A.

[0075] Experiments were performed to determine the purity of Form D (PSM1). With reference to Figure 26, the purity of Form D of ponatinib hydrochloride was determined to be 97.3664% (area percent).

[0076] Characteristics of E-form (HCl4+HCl1 mixture): HCl4 was obtained only as a mixture with Form A from grinding experiments with hexafluorobenzene (see overview in Figure 2).

[0077] Form E of ponatinib hydrochloride was found to be physically unstable when stored under ambient conditions. After 8 months of storage, the mixture HCl1 + HCl4 was re-examined by XRPD and had converted to Form A.

[0078] F-type (HCl5 class) characteristics: The HCl5 form was obtained from vapor diffusion experiments onto solids in butyl acetate as described herein (see overview in Figure 2). The HCl5 class was characterized by DSC, cycling DSC, TGMS, FTIR, HPLC, and DVS. Physical stability under short-term storage conditions (i.e., 1 week at 40°C and 75% RH) was investigated. Samples of the HCl5 class were physically stable after 8 months of storage under ambient conditions, as assessed by XRPD. After 1 week in a humidity chamber (40°C / 75% RH), the material was still the HCl5 class, although slight differences in the XRPD pattern were observed.

[0079] DVS experiments showed that HCl5 was highly hygroscopic, with a water mass adsorption rate of 37%. The material lost its crystallinity as seen by XRPD after the DVS experiment.

[0080] Form F was successfully scaled up to the 120 mg scale using the same conditions as the initial experiments identifying the previously discovered polymorphs. Two scale-up experiments were performed, and the corresponding XRPD patterns indicated that it was an isomorphous form of HCl5. These isomorphous forms, along with HCl5 and HCl5b, were designated as part of the HCl5 class, or Form F.

[0081] Figure 27 shows an XRPD overlay of (from bottom to top): HCl1 (Form A starting material); HCl5 and HCl5b (wet and dry VDS28, solvent: butyl acetate); HCl5-class (VDS1, solvent: butyl acetate), low-crystalline (VDS1 after DVS); HCl5-class (VDS2, solvent: butyl acetate); and HCl5-class (VDS2 after 1 week at 40°, 75% RH). In the XRPD pattern shown in Figure 27, HCl5 exhibits at least one or all of the following peaks at angles two-theta (2θ): 6.8; 9.8; 12.4; 16.2; 17.9; 19.0; 24.0; and 25.1. In certain embodiments, HCl5 is characterized by an XRPD pattern including one or more of the following peaks two-theta (2θ): 9.8; 12.4; and 25.1. In the XRPD patterns shown in Figure 27, the HCl5 class (top pattern) exhibits at least one or all of the following peaks at angles 2-theta (2θ): 7.9; 8.7; 9.7; 11.4; 15.6; 16.5; and 25.8. In certain embodiments, the HCl5 class is characterized by an XRPD pattern that includes one or more of the following peaks 2-theta (2θ): 15.6; 16.5; 25.8. In certain embodiments, the XRPD pattern of Form F exhibits two peaks, three peaks, four peaks, or five peaks.

[0082] Referring to Figure 28, the melting point of Form F of ponatinib HCl (PSM1) was determined by differential scanning calorimetry (DSC). Samples from two different experiments were analyzed in a crucible with a small hole, using a dry N2 gas purge, at a heating rate of 10°C / min over the temperature range of 25°C to 300°C. For one sample from one experiment (VDS1, top curve), the T peak = 120.7℃, T peak = 184.3°C and T peak A strong endothermic event was observed at T = 209.4 °C. In the other experimental sample (VDS2, bottom curve), peak = 122.1℃, T peak = 209.7°C and T peak A strong endothermic event was found to occur at 252.1°C.

[0083] Cyclic DSC experiments showed that when the HCl5 class desolvates, it melts at approximately 210°C and transforms into a form named "HCl5 desolvate."

[0084] Referring to Figure 29, the TGA / SDTA thermogram (VDS1, top) and TGMS (bottom) thermograms for Form F are shown. A mass loss of 17.1% (butyl acetate, API:solvent ratio 1:1.01) was observed over the temperature interval from 25°C to 160°C. TG-MS analysis revealed that the HCl5 class is a butyl acetate solvate with a 1:1 API:butyl acetate ratio, which desolvates at approximately 120°C. Figure 30 shows the corresponding TGA / SDTA (top) and TGMS (bottom) thermograms for Form F of VDS2. A mass loss of 16.6% (butyl acetate, API:solvent ratio 1:0.98) was observed over the temperature interval from 25°C to 160°C.

[0085] Regarding Figures 31 and 32, 1750-500 cm -1 The FT-IR spectrum of Form F is shown in the region of Figure 1. These data support the proposed structure of Form F of ponatinib hydrochloride. Furthermore, the spectrum shows unique features of Form F compared to Form A.

[0086] Experiments were performed to determine the purity of Form F (VDS2). With reference to Figure 33, the purity of Form D of ponatinib hydrochloride was determined to be 98.2833% (area percent).

[0087] Characteristics of G form (HCl5b): Form G of ponatinib hydrochloride was obtained by conversion of HCl5 by drying under full vacuum for 3 days. Form HCl5b was found to be physically stable even after 8 months of storage under ambient conditions.

[0088] Characterization data for Form G are presented herein in relation to Form F.

[0089] Characteristics of H-type (HCl6 class): HCl6 was obtained from two different experiments, evaporation into solution in MeOH / water and MeOH solvent systems, and evaporation onto solids, respectively (see overview in Figure 2). Differences in the time points at which the material was sampled resulted in slightly different corresponding XRPD patterns, indicating, without being bound by theory, that HCl6 is likely an isomorphous class form. HCl6 class was successfully scaled up to 120 mg using the same conditions as the original screening experiment, evaporation onto solids with MeOH.

[0090] The HCl6 class was characterized by DSC, cycling DSC, TGMS, FTIR, HPLC, and DVS. Physical stability under short-term storage conditions (i.e., 1 week at 40°C and 75% RH) was investigated. Samples of Form H were physically stable after 8 months of storage under ambient conditions as assessed by XRPD. After 1 week in a humidity chamber (40°C / 75% RH), the material was still HCl6 class, but slight differences were observed in the XRPD.

[0091] Figure 34 shows an overlay of the XRPD patterns for (from bottom to top): Form A (ponatinib hydrochloride starting material), HCl6-class (VDS6, solvent: methanol), HCl6-class (VDS3, solvent: methanol), HCl6 (VDS3 after DVS), HCl6-class (VDS3 after climate chamber), HCl6-class (VDS4, solvent: methanol), and HCl6-class (VDS4 after DVS). The XRPD pattern shown in Figure 34 shows at least one or all of the following peaks at angles two-theta (2θ) for HCl6 (immediately above the Form A pattern): 5.9; 8.1; 9.5; 10.7; 13.4; 16.0; 17.0; 22.0; 22.8; 24.7; and 28.3. In certain embodiments, HCl6 is characterized by an XRPD pattern that includes one or more of the following peaks at 2-theta (2θ): 8.1; 10.7; 13.4; 24.7; and 28.3. In the XRPD patterns shown in Figure 34, the HCl6 class (top pattern) exhibits at least one or all of the following peaks at angles 2-theta (2θ): 8.0; 10.2; 10.9; 11.8; 14.1; 15.4; 16.3; 19.9; 22.3; 23.7; 25.0; and 28.2. In certain embodiments, the HCl6 class is characterized by an XRPD pattern that includes one or more of the following peaks at 2-theta (2θ): 10.2; 15.4; 23.7; 25.0. In certain embodiments, the XRPD pattern of Form F exhibits two peaks, three peaks, four peaks, or five peaks. XRPD analysis of both samples showed that almost the same pattern was observed after DVS, however TGMS analysis of VDS4 showed that there were no more methanol molecules in the sample, replaced by water molecules forming what appeared to be a hemihydrate form belonging to the HCl6 class.

[0092] Referring to Figure 35, the melting point of Form H of ponatinib HCl was determined by differential scanning calorimetry (DSC). Samples from two different experiments were analyzed in a crucible with a small hole, using a dry N gas purge, at a heating rate of 10°C / min over the temperature range of 25°C to 300°C. For one experiment (VDS3, top curve), the T peak The other experiment (VDS4, bottom curve) revealed a strong endothermic event at T = 219.4 °C. peak = 219.4°C and T peak A strong endothermic event was found to occur at 256.8°C.

[0093] With reference to Figure 36, the TGA / SDTA thermogram (VDS3, top) and TGMS (VDS3, bottom) thermograms for Form H are shown. A 5.4% mass loss (methanol, API:solvent ratio 1:1.01) was observed in the temperature interval from 30°C to 150°C, and a 0.3% mass loss (methanol, API:solvent ratio 1:0.05) was observed in the temperature interval from 190°C to 220°C. For VDS4, the corresponding TGA / SDTA thermogram (top) and TGMS (bottom) thermograms for Form H are shown in Figure 37. A 3.3% mass loss (methanol, API:solvent ratio 1:0.6) was observed in the temperature interval from 30°C to 150°C, and a 0.7% mass loss (methanol, API:solvent ratio 1:0.12) was observed in the temperature interval from 190°C to 220°C.

[0094] Regarding Figures 38 and 39, 1750-500 cm -1 The FT-IR spectrum of Form H of ponatinib hydrochloride is shown in the region of Figure 1. These data support the proposed structure of Form H of ponatinib hydrochloride. Furthermore, the spectrum shows unique features of Form H compared to Form A.

[0095] Experiments were performed to determine the purity of Form H (VDS4). With reference to Figure 40, the purity of Form H of ponatinib hydrochloride was determined to be 97.9794% (area percent).

[0096] Characteristics of Form I (HCl6 desolvate): Cyclic DSC experiments carried out in conjunction with the H-form experiments showed that upon desolvation, the HCl6 class transforms into a form designated "HCl6-desolvate," which melts at approximately 220°C.

[0097] Characteristics of J-form (HCl7): Form J is the pentahydrate of ponatinib HCl and was discovered in conjunction with single crystal analysis. Form J is the most stable hydrated structure identified, as indicated by competitive water slurry between the trihydrate and pentahydrate.

[0098] Single crystals of suitable size were obtained in vapor diffusion experiments carried out in the solvent mixture methanol / water (20:80) with n-butyl acetate as the antisolvent. One parallelepiped single crystal measuring approximately 0.45 × 0.25 × 0.12 was collected from the crystallization vial and mounted on a glass fiber. Its crystallographic data (collected up to θ = 27.5°) are listed in Table 4.

[0099] [Table 5]

[0100] The asymmetric unit contains a cation, a chloride anion, and five water molecules (pentahydrate). The water molecules are bound to the anion, cation, and neighboring water molecules via hydrogen bonds (H-bonds).

[0101] An important consequence of the parent H-bonding arrangement is that in this crystal both charged atoms (i.e., the protonated nitrogen of the API and the chloride anion) are bridged / separated by multiple water molecules.

[0102] Figure 43 shows a characteristic X-ray powder diffraction (XRPD) pattern of Form J of ponatinib hydrochloride. The XRPD pattern of Form J shown in Figure 43 exhibits at least one or more of the following peaks at angles two-theta (2θ) with a relative intensity of 20% or greater: 6.1; 7.0; 13.3; 16.4; 20.7; 22.2; 23.9; 25.5; and 29.1. In certain embodiments, Form J is characterized by an XRPD pattern including one or more of the following peaks two-theta (2θ): 7.0; 22.2; and 25.5. In certain embodiments, the XRPD pattern of Form J exhibits two peaks, three peaks, four peaks, or five peaks.

[0103] Characteristics of K-type (HCl8): Form K was discovered in connection with single crystal analysis. Single crystals were grown in gentle evaporation experiments carried out in a 50:50 TFE / H2O mixture. A bulk single crystal measuring approximately 0.40 x 0.30 x 0.25 mm was analyzed. Although the crystal was large, it diffracted very poorly, indicating irregularities in the structure. Therefore, measurements were recorded only up to θ = 25°. Crystallographic parameters are listed in Table 5.

[0104] [Table 6]

[0105] The structure of the mixed TFE solvate / hydrate form contains a cation, a chloride anion, and two neutrals, trifluoroethanol and a water molecule. In this structure, the water molecule participates in H-bonding but does not separate the charged atoms, as in the pentahydrate and trihydrate forms. TFE and the water molecule act only as donors in the hydrogen-bonding network. In particular, only one of the hydrogen atoms in the water molecule acts as a donor, which is thought to be the reason for the disordered water molecule and the non-stoichiometric ratio of water molecules to API molecules.

[0106] Figure 44 shows a characteristic X-ray powder diffraction (XRPD) pattern of Form K of ponatinib hydrochloride. The XRPD pattern of Form K shown in Figure 44 exhibits at least one or more of the following peaks at angles two-theta (2θ) with a relative intensity of 20% or greater: 6.1; 7.4; 13.5; 17.4; 18.5; 20.7; 23.9; and 28.3. In certain embodiments, Form K is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 7.4 and 23.9. In certain embodiments, the XRPD pattern of Form K exhibits two peaks, three peaks, four peaks, or five peaks.

[0107] Characteristics of amorphous ponatinib hydrochloride: Figure 45 shows the XRPD patterns of Form A of ponatinib hydrochloride (lower pattern) and amorphous ponatinib hydrochloride (upper pattern) (solvent: 2,2,2-trifluroethanol). It can be readily seen that Form A exhibits a distinct set of peaks at specific angles 2-theta, while amorphous ponatinib hydrochloride lacks any distinct peaks.

[0108] Additionally, amorphous ponatinib hydrochloride has a unique melting temperature compared to Form A of ponatinib hydrochloride. Figure 46 shows a characteristic differential scanning calorimetry (DSC) thermogram of amorphous ponatinib hydrochloride. A strong endothermic event was observed with a peak at 259.4°C, which corresponds to the melting point of the amorphous form. This melting point is different from that observed for Form A of ponatinib hydrochloride, which exhibited a melting point of 264.1°C.

[0109] The unique and different physical properties of amorphous ponatinib hydrochloride and Form A of ponatinib hydrochloride are not believed to be attributable to the purity of each material. In the case of amorphous ponatinib hydrochloride, HPLC revealed a purity of 99.7877% (area percent) material (see Figure 47), while Form A of ponatinib hydrochloride was found to be 99.8% (area percent). Example

[0110] Example 1 Polymorphic Discovery Initial efforts to discover polymorphs of ponatinib hydrochloride were divided into two phases. Phase 1 included characterization of the starting material, feasibility studies, and solubility studies to provide data for solvent selection in Phase 2. Phase 2 included 192 milliliter (ml)-scale polymorph screening experiments. Eight polymorphs, Forms A, B, C, D, E, F, G, and H, were discovered from these initial efforts.

[0111] Phase 1: Characterization of starting materials Approximately 24 grams of the compound ponatinib hydrochloride was obtained as a pale yellow solid. This starting material was characterized by XRPD, digital imaging, DSC, TGMS, and HPLC. The starting material, 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride, was obtained as a crystalline material (designated HCl1), the chemical purity of which was estimated to be 99.8% by HPLC. TGA and TGMS analysis showed a mass loss of 0.7% (residual ethanol) between 25°C and 240°C prior to the thermal decomposition process. DSC analysis showed that T peak = 264.8 °C, which is likely due to melting and / or decomposition of the compound, 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride.

[0112] Stage 1: Solubility test A quantitative solubility study of the ponatinib hydrochloride starting material was conducted using a set of 20 solvents. After preparing slurries with an equilibration time of 24 hours, the slurries were filtered. Solubility was determined from saturated solutions by HPLC. The residual solids were characterized by XRPD. The results are summarized in Table 6.

[0113] [Table 7]

[0114] In 19 of the experiments shown in Table 6, the material analyzed after solubility evaluation in 19 different solvents appeared to be the same form as the starting material, designated HCl1. Experiment QSA13, performed with 2,2,2-trifluoroethanol, resulted in complete dissolution of the material at the selected concentration, and the sample obtained after solvent evaporation yielded amorphous material. The solids obtained from two water slurries (QSA12 and QSA21) each yielded two distinct forms: Form HCl2 and Form HCl2b. After several days of storage under ambient conditions, Form HCl2 converted to Form HCl2b and could not be further characterized. Further characterization revealed that Form HCl2b was a hydrated form (API / water ratio 1:1.4).

[0115] Phase 1: Feasibility testing A feasibility study was conducted to attempt to obtain an amorphous starting material that could be used in several crystallization techniques for the second part of this study. Two techniques were used: milling and freeze-drying. The results are shown below.

[0116] crushing Two grinding experiments were performed at a frequency of 30 Hz for different durations. After 60 minutes of grinding, the crystalline starting material was converted to amorphous form. After 120 minutes, the resulting material remained amorphous and had a chemical purity of 99.6%.

[0117] Freeze drying Eight lyophilization experiments were performed with 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride and are summarized in Table 7.

[0118] [Table 8]

[0119] The solubility of the compound ponatinib hydrochloride in tetrahydrofuran, 2-methyltetrahydrofuran, and dichloromethane was too low to allow for lyophilization under favorable conditions. Solvents such as methanol, 2,2,2-trifluoroethanol (TFE), and TFE / water mixtures yielded amorphous material. Samples obtained from neat TFE or solvent mixtures with a high TFE content contained 11% residual solvent in the dried powder (TGMS results). Samples obtained from methanol and TFE / water (50:50) contained less residual solvent, only 0.9% and 1.5%, respectively. Further drying for 24 hours reduced the amount of residual solvent in the amorphous material produced from TFE / water (50:50) to less than 1%. The chemical purity of both the amorphous samples obtained from methanol and TFE / water (50:50) was estimated to be 99.8% by HPLC. Because creeping was observed in lyophilization experiments using methanol, a TFE / water (50:50) method was selected to generate amorphous ponatinib hydrochloride for use in second-stage cooling-evaporation crystallization and vapor diffusion onto solid experiments.

[0120] Phase 2: Polymorphism Discovery Polymorph screening experiments of ponatinib hydrochloride were conducted at the milliliter (ml) scale using 192 different conditions using six different crystallization methods: (1) cooling-evaporation; (2) antisolvent addition; (3) grinding; (4) slurry; (5) vapor diffusion into solution; and (6) vapor diffusion onto solid. After the screening experiments were completed, the material was collected and analyzed by XRPD and digital imaging.

[0121] Cooling-evaporation crystallization experiment Thirty-six ml-scale cryo-evaporation experiments, shown in Table 8, were performed in 1.8 ml vials using 36 different solvents and solvent mixtures and one concentration. 25 mg of amorphous ponatinib hydrochloride was weighed into each vial. Screening solvent was then added until a concentration of approximately 60 mg / ml was reached. Additionally, vials containing magnetic stir bars were capped and placed in an Avantium Crystal 16 to run a temperature profile (as described in Table 9 below). The mixtures were cooled to 5°C and held at that temperature for 48 hours, after which the vials were placed under vacuum. The solvent was allowed to evaporate at 200 mbar or 10 mbar for several days and then analyzed by XRPD and digital imaging.

[0122] [Table 9]

[0123] [Table 10]

[0124] Crash crystallization experiments with antisolvent addition The rapid crystallization experiments used 36 different crystallization conditions using one solvent and 24 different anti-solvents (see Table 9). Anti-solvent addition experiments proceeded as follows: A stock solution was prepared, and after 24 hours of equilibration, the concentration of ponatinib hydrochloride reached its saturated concentration at ambient temperature and was then filtered into 8 ml vials. A different anti-solvent was added to each of these vials using a solvent to anti-solvent ratio of 1:0.25. Since no precipitation occurred, this ratio was increased to 1:4 with a 60 minute wait time between additions. Since there was still no precipitation, the solvent was completely evaporated under vacuum at room temperature. After evaporation, this experiment resulted in no yield.

[0125] [Table 11]

[0126] Crushing experiment In the droplet milling method, a small amount of solvent was added to the starting material, 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride, and the mixture was milled in a stainless steel milling jar containing two stainless steel milling balls. In this manner, the effects of 24 different solvents (see Table 10) were investigated. Typically, 30 mg of starting material was weighed into a milling vessel, and 10 μL of solvent was added to the vessel. Milling experiments were carried out at 30 Hz for 120 minutes. Each wet material was then analyzed by XRPD and digital imaging.

[0127] [Table 12]

[0128] Slurry experiments A total of 48 slurry experiments were conducted using the compound ponatinib hydrochloride and 24 different solvents at 10°C and 30°C over a two-week period. Table 11 summarizes the experimental conditions. The experiments were conducted by stirring the suspension of the material in the solvent at a controlled temperature. At the end of the slurrying period, the vials were centrifuged to separate the solid from the mother liquor. The solid was further dried under full vacuum at room temperature and analyzed by XRPD and digital imaging.

[0129] [Table 13]

[0130] Vapor diffusion into solution In this vapor diffusion experiment, a saturated solution of ponatinib hydrochloride was exposed to solvent vapors at room temperature for two weeks. A volume of the saturated solution was transferred to an 8 ml vial, left open, and placed in a closed 40 ml vial containing 2 ml of antisolvent (see Table 12). After two weeks, the sample was observed to form a solid. Drying the sample under vacuum (200 mbar or 10 mbar) resulted in no yield. Based on this result, new experiments were performed with 12 different crystallization conditions as listed in the table (Experiment IDs, VDL25–VDL36).

[0131] [Table 14]

[0132] Vapor diffusion onto solids In this vapor diffusion experiment, amorphous ponatinib hydrochloride was exposed to solvent vapors at room temperature for two weeks. An 8 ml vial containing amorphous API was left open and placed in a closed 40 ml vial containing 2 ml of antisolvent (see Table 13). After two weeks, the solid was analyzed by XRPD and digital imaging. If the solid had liquefied due to vapor, the sample was dried under vacuum (200 mbar or 10 mbar) before being analyzed by XRPD and digital imaging.

[0133] [Table 15]

[0134] In these initial crystallization experiments, XRPD analysis of the resulting dried (and wet, where applicable) samples revealed the presence of seven new polymorphs in addition to the amorphous material and the starting Form A. The seven forms are designated HCl2, HCl2b, HCl3-class, HCl5, HCl5b, HCl6-class, and the mixture HCl1+HCl4.

[0135] The occurrence of the various forms obtained in the second phase of these initial efforts is shown in Figure 2. Representative XRPD patterns and digital images of each form obtained in these second phase experiments were obtained. Characterization of the forms obtained in the first phase of these initial efforts is summarized in Table 14.

[0136] [Table 16]

[0137] The polymorphs identified in these Phase 1 and Phase 2 experiments and shown in Figure 2 were primarily assigned by XRPD analysis. During this analysis, it was observed that some patterns had nearly identical general fingerprints of the XRPD patterns but exhibited some slight differences, such as peak shifts or additional, smaller peaks. These types of patterns were grouped together as pattern classes (e.g., HCl class). Based on XRPD, it was concluded that the similarities between XRPD patterns within a class can be explained by the fact that these solid forms are isomorphous hydrates / solvates (crystal packing is nearly identical, but slight differences in unit cell parameters are observed due to the incorporation of different solvents and water into the crystal structure).

[0138] Isomorphous solvate classes were designated by numbers (HCl3 class) or number-letter combinations (e.g., HCl2 and HCl2b). Isomorphous solvate / hydrate classes designated by letter-number combinations indicate that several subclasses of this class were observed experimentally (e.g., HCl2 and HCl2b). When four or more subclasses within a class were identified, all XRPD patterns corresponding to the isomorphous solvate / hydrate class were regrouped under a single number (e.g., HCl3 class).

[0139] Isomorphous solvates within a particular class or between classes with the same numerical designation exhibit greater similarity in their XRPD patterns than do isomorphous solvate classes with different numerical designations. The greater differences in the XRPD patterns of these different classes of isomorphous hydrates / solvates reflect significantly different packings in the crystal structures.

[0140] Some XRPD patterns had one or two additional peaks observed compared to the identified forms, and these were designated "plus peaks" because they could not be unambiguously assigned to known forms.

[0141] Example 2 Further discovery of polymorphs Subsequent efforts were undertaken to analyze single crystals of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride. These efforts led to the discovery of five different pseudopolymorphs, two of which were previously undiscovered. The two newly discovered polymorphs are designated herein as HCl7 (also referred to herein as "Form J") and HCl8 (also referred to herein as "Form K"). In these subsequent experiments, three different crystallization methods were used to grow single crystals of a size suitable for analysis: (1) gentle evaporation of the crystallization solvent; (2) antisolvent diffusion into 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride solution; and (3) temperature-controlled crystallization. In these subsequent experiments, a total of 54 crystallization experiments were performed in an attempt to grow single crystals of the hydrated form of ponatinib hydrochloride for structure determination.

[0142] For temperature-controlled crystallization, 24 experiments were prepared using alcohol and water mixtures (see Table 15). Each experiment contained 10 mg of ponatinib hydrochloride. The API and solvent mixture was rapidly heated up to 80°C and slowly cooled (0.1°C / min) to room temperature.

[0143] [Table 17]

[0144] Twenty-five experiments were performed on evaporation into solution. In each experiment, 10 mg of ponatinib hydrochloride was dissolved in 1 ml of a TFE / water (10:90) mixture or a MeOH / water (30:70) mixture. Each solution was placed in a 6 ml vial, which was then placed in a 20 ml vial containing 3 ml of antisolvent. The vials were stored at room temperature for 2 to 4 weeks. Details are reported in Table 16.

[0145] [Table 18]

[0146] For gentle evaporation of the solvent, 10 mg of ponatinib hydrochloride was placed in an 8 ml vial and 2 ml of solvent (mixture of solvents) was added. If the solid did not dissolve, the vial was heated to 90° C. The mixture was then allowed to cool gently to room temperature (see Table 17). For Experiment 53, listed at the end of Table 17, the material did not completely dissolve even after being held at 90° C. for several hours.

[0147] [Table 19]

[0148] Each polymorph disclosed herein is produced by a specific crystallization / solvent method using ponatinib HCl as the starting material. The synthesis of ponatinib HCl has been previously described (e.g., WO 2007 / 075869 and WO 2011 / 053938), but the following synthesis of ponatinib HCl is described in Example 6.

[0149] Example 3 Stress testing of form A of ponatinib hydrochloride Form A is a crystalline anhydrous solid obtained with high reproducibility from a variety of solvents. Form HCl-1 is inherently chemically stable, which directly correlates with the thermodynamic stability of Form HCl-1. Form HCl-1 is stable to temperature, pressure, and humidity stresses, as well as exposure to some solvent vapors, and is thermodynamically stable. Numerous studies have been conducted to confirm its stability in both the formulated (tablet) and unformulated (drug substance) forms. The results of these studies are listed below in Table 18.

[0150] [Table 20]

[0151] Form HCl-1 is stable to temperature, pressure and humidity stresses, as well as exposure to solvent vapors, and is the most thermodynamically stable solid form isolated to date.

[0152] To test the physical stability of crystalline Form HCl1, an experiment was performed as follows.

[0153] Crystalline HCl form 1 and a 50:50 physical mixture of HCl form 1 and amorphous material were exposed to ethanol vapor for 2 weeks (see vapor diffusion experiments). 2 and 100kN / cm 2 (or 4 tonnes / cm 2 and 8 tonnes / cm 2Tablets were prepared by applying a pressure of 0.1 MPa for 10 seconds. Form A was stored in capsules under ambient conditions for up to 17 months. These samples were analyzed by high-resolution XRPD.

[0154] The results obtained are summarized in Table 19, and XRPD measurements and digital images were also obtained, which showed that the HCl1 polymorph remained unchanged under the range of applied stress conditions, confirming its excellent physical stability.

[0155] [Table 21]

[0156] Example 4 Stability of specific polymorphs Samples of eight solid forms of the HCl salt were selected to test their physical stability. Two representative samples of each relevant polymorph of the resulting HCl salt were selected. Each sample was re-analyzed by XRPD. The physical stability of each form after 8 months of storage under ambient conditions. The results are summarized below: HCl1, HCl2b, HCl3 class, HCl5b and HCl6 class are stable under the conditions studied; HCl2 was converted to HCl2b (this conversion occurred already after storing the sample under ambient conditions for 1 day); HCl5 was converted to HCl5b (this conversion occurred already after drying under full vacuum for 3 days); The mixture HCl1+HCl4 was converted to HCl1 after 8 months under ambient conditions.

[0157] [Table 22]

[0158] Example 5 Preparation of Form A Form A of ponatinib HCl is formed as a crystalline material by adding an ethanolic solution of HCl (1.0 equivalent) to an ethanolic solution of ponatinib free base. The drug substance, ponatinib HCl, is crystallized during the final stage of drug substance synthesis by the addition of seed crystals, which results in a highly consistent and characteristic particle size and range for the drug substance. The ethanol content of the final 10 kilogram-scale batches of ponatinib HCl Form 1 ranged from 0.8 to 1.2%.

[0159] Form HCl-1 showed no evidence of ethanol or water, indicating that Form A is anhydrous. Furthermore, the crystal packing of Form HCl-1 lacks void spaces that could accommodate small organic molecules, including ethanol. New studies examining the ethanol content and removal of ethanol from ponatinib HCl during drying revealed that ethanol appears to be bound to the crystalline surface of Form A ponatinib HCl.

[0160] The HCl-1 form is characterized by the consistent presence of residual ethanol in all batches of the drug substance at levels of approximately 1% by weight. Crystallographic and other studies have shown that the residual ethanol is present on the crystal surface (trapped) and not part of the crystalline unit cell, and that HCl-1 is not an ethanol solvate or channel solvate. The last 10 multikilogram-scale drug substance batches had ethanol levels ranging from 0.8 to 1.2%.

[0161] II. Polymorphs of Ponatinib Free Base The preparation of the ponatinib free base starting material (AP23534) is illustrated and described in the Synthesis section of this disclosure (see Scheme 1). The preparation of the amorphous free base is described later in this disclosure. "Feasibility study of ponatinib free base compound" This is discussed in the section entitled:

[0162] XRPD analysis has discovered a total of 11 polymorphs of ponatinib, including anhydrate, hydrate, solvate, and isomorphous hydrate / solvate forms, originating from the ponatinib free base starting material. These 11 novel polymorphs are referred to herein as Form A, Form B (or B-class form), Form C, Form D, Form E (or E-class form), Form F, Form G, Form H (or H-class form), Form I, Form J, and Form K.

[0163] Anhydrous Form A (melting point around 200 °C) was the major crystalline form observed in screening. Form B includes four isomorphous solvates: a 1:1 dioxane solvate, a 1:1 perfluorobenzene solvate, a 1:0.4 2-methyl THF solvate, and a 1:1 cyclohexanone solvate. Form E includes chloroform and dichloromethane isomorphous solvates. Form D is a solvate form (e.g., DMA). Form F is a monohydrate form. Form H includes two isomorphous solvates: a 1:0.6 1-propanol solvate and a 1:0.93 2-methoxyethanol solvate.

[0164] The assignment of forms obtained in the third and fourth stage experiments was primarily based on XRPD analysis. From this analysis, it could be observed that some patterns had almost the same general fingerprint of the XRPD pattern, but showed some slight differences such as peak shifts and / or additional minor peaks. These types of patterns were grouped together as a pattern class (e.g., B class). Based on XRPD, the similarity between XRPD patterns within a class may indicate that these solid forms are isomorphous hydrates / solvates (crystal packing is almost the same, but there are slight differences in unit cell parameters due to the incorporation of different solvents and water into the crystal structure). The classes of isomorphous solvates were designated by letters (B class).

[0165] Some XRPD patterns contained one or more additional peaks compared to the various identified forms. These peaks could not always be unambiguously assigned to known forms and were therefore designated as "plus peaks" where appropriate (e.g., "Form F plus peak").

[0166] The polymorph screening of ponatinib involved six crystallization methods performed on a milliliter scale. These methods included: rapid crystallization by antisolvent addition; milling; slurry experiments; vapor diffusion into solution; cooling-evaporation crystallization; and vapor diffusion onto solid. These methods are described above for the preparation of the various ponatinib hydrochloride polymorphs. Any differences in the methods as applied to the ponatinib free base polymorphs are noted below.

[0167] The crystal structures of Form A and the two solvates of Form B were determined by single crystal X-ray data analysis, which revealed the conformation adopted by Form A in the crystal structure and the solvent content in the two solvates of Form B.

[0168] Generally, these various crystalline forms of ponatinib disclosed herein have advantageous physical properties (e.g., increased stability) for solid dosage commercial formulations compared to amorphous ponatinib. Using the same types of physicochemical data (e.g., DSC, XRPD, thermal analysis) used to distinguish the individual crystalline forms of ponatinib disclosed herein, the differences between crystalline ponatinib and amorphous ponatinib can be readily discerned.

[0169] Figures 48 and 49 tabulate 10 of the 11 solid state forms of ponatinib, including the polymorphs and pseudopolymorphs identified as Forms A through J disclosed herein (Form K is discussed separately later in this specification and is not part of this summary table). Figures 48 and 49 summarize the origin, occurrence, and various characteristics of the ponatinib polymorphs.

[0170] With reference to Figure 48, "LC" refers to low-crystalline ponatinib (e.g., Form C), which is used as the starting material for the various other polymorphs. "A" refers to Form A, which is the most abundant ponatinib crystalline form and can be used to prepare the other forms by the various crystallization methods listed above. The footnotes appearing in the table of Figure 48 are as follows: (a) Classification by XRPD after the crystallization experiment is completed; (b) Crystallization methods: cooling-evaporation (PSM), rapid crystallization with antisolvent addition (AS), grinding (GRP), slurry (SLP), vapor diffusion onto solid (VDS) and vapor diffusion into solution (VDL). Freeze drying (FD) was used to produce poorly crystalline material (see experiments in Phase 3). QSA: quantitative solubility experiments (Phase 3); (c) Occ: Total occurrence includes 192 experiments performed in the fourth stage, in which 61 samples were further analyzed wet or the mother liquor was evaporated and analyzed, resulting in a total of 253 substances characterized. For example, "(6, 2.4%)" corresponds to the form appearing in 6 of the 253 measurements, representing a percentage of 2.4%. For 4 of the 253 measurements (1.6%), the yield or scattering intensity of some products was too low to identify the solid form, or the substance was wet; (d) PO: preferred orientation effect; and (e) Starting material: Form A or low crystalline (LC) material obtained by lyophilization.

[0171] The above methodology used to assess ponatinib hydrochloride polymorphism is described below for polymorphism of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide free base.

[0172] Characteristics of Form A ponatinib Anhydrous Form A (the same crystalline form as the starting material) was the predominant crystalline form discovered in the screening. The chemical structure of ponatinib Form A has been unambiguously established by single crystal X-ray crystallography. The solid form of ponatinib observed is Form A, an anhydrous crystalline solid.

[0173] Figure 50 shows the molecular structure and numbering scheme of Form A of ponatinib free base compound (anhydrous) obtained from single crystal X-ray diffraction. Based on this structural analysis, Form A was found to be the anhydrous form.

[0174] Crystallographic data (collected up to θ=26°) for anhydrous Form A are listed in Table 21.

[0175] [Table 23]

[0176] Figure 51 shows a comparison of the experimental XRPD pattern of Form A with the calculated pattern (assuming FWHM=0.1°) based on the determined crystal structure. The close similarity of the two XRPD patterns indicates that the crystal structure of Form A is representative of the bulk material.

[0177] In the XRPD pattern shown in Figure 51, crystalline Form A exhibits at least one or all of the following peaks at angles two theta (2θ): 6.2; 8.8; 9.9; 11.2; 12.3; 12.9; 13.5; 13.8; 14.2; 14.4; 16.0; 16.4; 17.2; 17.6; 18.0; 18.2; 19.3; 19.5; 19.8; 20.6; 21.5; 21.9; 22.2; 22.6; 23.1; 24.0; 24.4; 25.1; 25.6; 25.9; 26.8; 27.4; 27.8; 29.1; and 29.8. In certain embodiments, Form A is characterized by an XRPD pattern that includes one or more of the following peaks two theta (2θ): 6.2; 12.3; 13.8; 14.4; 16.0; 16.4; 17.2; 17.6; 18.2; 19.5; 19.8; 20.6; 21.5; 22.2; 24.0; 25.9; 26.8; 27.4; and 27.8. In the XRPD pattern shown in Figure 51, Form A exhibits at least one or all of the following peaks at angles two theta (2θ): 12.3; 13.8; 14.4; 17.6; 19.8; 20.6; 21.5; 22.6; and 24.0. In certain embodiments, Form A is characterized by an XRPD pattern comprising one or more of the following peaks 2-theta (2θ): 12.3; 13.8; 17.6; 19.8; 20.6; 21.5; 22.6; and 24.0. In certain embodiments, the XRPD pattern of Form A exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, crystalline Form A of ponatinib free base is characterized by an XRPD pattern substantially similar to the upper XRPD pattern (obtained pattern) in Figure 51. In certain embodiments, crystalline Form A comprises an XRPD pattern having characteristic peaks expressed in degrees 2-theta as shown in either the lower (simulated pattern) or upper (obtained pattern) pattern in Figure 51.

[0178] Referring to Figure 52, the melting point of anhydrous Form A of ponatinib was determined by differential scanning calorimetry (DSC). Two samples of ponatinib, AP24534 Lot 1285.206A (treated with 1-PrOH) and AP24524 Lot 1-PrOH untreated, were analyzed in small-hole crucibles with a dry N gas purge at a heating rate of 10°C / min over the temperature range of 30°C to 350°C. An endothermic event was observed with a peak at 199.6°C, corresponding to the melting point of Form A of ponatinib.

[0179] Characteristics of the B-type (B-class) ponatinib polymorph Characteristics of ponatinib / 1,4-dioxane 1:1 solvate form B Single crystal X-ray diffraction analysis was used to determine the crystal structure of the 1:1 1,4-dioxane solvated Form B. Figure 53 shows the molecular structure and numbering scheme of the 1:1 1,4-dioxane solvated Form B obtained from single crystal X-ray diffraction.

[0180] Crystallographic data (collected up to θ=27.4°) for the 1:1 1,4-dioxane solvate Form B are listed in Table 22.

[0181] [Table 24]

[0182] Figure 54 shows the calculated pattern of ponatinib / 1,4-dioxane 1:1 solvate Form B based on the determined crystal structure.

[0183] In the XRPD pattern shown in Figure 54, ponatinib / 1,4-dioxane 1:1 solvate Form B exhibits at least one or all of the following peaks at angles two theta (2θ): 5.6; 7.2; 9.8; 10.8; 12.1; 12.5; 12.8; 14.5; 15.3; 15.8; 17.0; 17.3; 17.5; 18.5; 19.0; 19.5; 20.0; 20.3; 21.1; 21.6; 22.4; 22.8; 23.5; 24.1; 24.5; 25.3; 26.0; 26.4; 27.0; 27.5; 28.4; 30.8; and 32.0. In certain embodiments, ponatinib / 1,4-dioxane 1:1 solvate Form B is characterized by an XRPD pattern that includes one or more of the following peaks two-theta (2θ): 5.6; 10.8; 12.1; 14.5; 15.3; 15.8; 17.0; 17.3; 18.5; 19.0; 20.0; 20.3; 21.6; 22.4; 24.5; and 26.0. In the XRPD pattern shown in Figure 54, ponatinib / 1,4-dioxane 1:1 solvate Form B exhibits at least one or all of the following peaks at angles two-theta (2θ): 5.6; 12.1; 14.5; 15.3; 15.8; 17.3; 18.5; 19.0; 20.3; 21.6; and 26.0. In certain embodiments, ponatinib / 1,4-dioxane 1:1 solvate Form B is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 5.6; 14.5; 15.3; 15.8; 19.0; 20.3; and 26.0. In certain embodiments, the XRPD pattern of ponatinib / 1,4-dioxane 1:1 solvate Form B exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, ponatinib / 1,4-dioxane 1:1 solvate Form B is characterized by an XRPD pattern substantially similar to the XRPD pattern in Figure 54. In certain embodiments, ponatinib / 1,4-dioxane 1:1 solvate Form B is crystalline.

[0184] Characterization of ponatinib / perfluorobenzene 1:1 solvate form B The crystal structure of the 1:1 perfluorobenzene solvate Form B was determined using single crystal X-ray diffraction analysis. Figure 55 shows the molecular structure and numbering scheme of the 11:1 perfluorobenzene solvate Form B obtained from single crystal X-ray diffraction. The similarity of the unit cell and crystal packing between the 1,4-dioxane solvate and the fluorobenzene solvate confirmed that these Form B class materials are isomorphous solvates.

[0185] Crystallographic data (collected up to θ=27.6°) for the 1:1 perfluorobenzene solvate Form B are listed in Table 23.

[0186] [Table 25]

[0187] Figure 56 shows a comparison of the experimental XRPD pattern of Form B 1:1 perfluorobenzene solvate with the calculated pattern based on the determined crystal structure (assuming FWHM = 0.1 °C). The close similarity of the two XRPD patterns indicates that the crystal structure of Form B 1:1 perfluorobenzene is representative of the bulk material.

[0188] In the XRPD pattern of Figure 56, Form B 1:1 ponatinib / perfluorobenzene solvate exhibits at least one or all of the following peaks at angles two theta (2θ): 7.0; 8.2; 9.8; 11.0; 11.5; 12.4; 12.8; 124.0; 14.4; 15.3; 16.0; 16.6; 17.2; 18.2; 19.0; 19.5; 20.0; 20.1; 21.1; 22.0; 22.5; 22.7; 23.5; 24.0; 24.5; 25.1; 26.0; 27.0; 27.8; 28.2; 31.8; and 35.4. In certain embodiments, Form B of 1:1 ponatinib / perfluorobenzene solvate is characterized by an XRPD pattern that includes one or more of the following peaks two-theta (2θ): 5.6; 11.0; 12.4; 12.8; 14.4; 15.3; 16.0; 16.6; 17.2; 18.2; 19.0; 20.1; 22.0; 22.5; 22.7; 23.5; 24.5; 26.0; and 28.2. In the XRPD pattern shown in Figure 56, Form B of 1:1 ponatinib / perfluorobenzene solvate exhibits at least one or all of the following peaks in degrees two-theta (2θ): 5.6; 12.4; 15.3; 16.0; 19.0; 20.1; 22.0; 22.5; 22.7; and 26.0. In certain embodiments, Form B of 1:1 ponatinib / perfluorobenzene solvate is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 5.6; 12.4; 15.3; 22.0; and 26.0. In certain embodiments, the XRPD pattern of Form B of 1:1 ponatinib / perfluorobenzene solvate exhibits two peaks, three peaks, four peaks, or five peaks. In certain embodiments, Form B of 1:1 ponatinib / perfluorobenzene solvate is characterized by an XRPD pattern substantially similar to the top XRPD pattern in Figure 56. In certain embodiments, Form B of 1:1 ponatinib / perfluorobenzene solvate is characterized by an XRPD pattern substantially similar to the bottom XRPD pattern in Figure 56. In certain embodiments, crystalline Form B comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees two-theta as shown in either the top or bottom of Figure 56.

[0189] Characterization of ponatinib / cyclohexanone 1:1 solvate form B and ponatinib / 2-methylTHF 1:0.4 solvate form B . Figure 57 shows a comparison of the XRPD patterns of two other B-class forms with the XRPD pattern obtained for Form A. The top pattern is obtained with a 1:0.4 ponatinib / 2-methyl THF solvate (B-class, lyophilized, solvent=2-methyl THF), GEN8.1. The middle pattern is obtained with a 1:1 ponatinib / cyclohexanone solvate (B-class, solvent=cyclohexanone), QSA7.1. The bottom pattern is obtained with anhydrous Form A.

[0190] In the above XRPD pattern shown in Figure 57, Form B 1:0.4 ponatinib / 2-methyl THF solvate exhibits at least one or all of the following peaks at angles two theta (2θ): 5.6; 9.5; 10.5; 11.2; 12.0; 12.5; 13.5; 14.0; 15.0; 15.5; 16.2; 16.8; 17.0; 18.1; 18.8; 20.1; 21.2; 22.1; 26.0; 26.1; 27.5; and 28.2. In certain embodiments, Form B of 1:0.4 ponatinib / 2-methyl THF solvate is characterized by an XRPD pattern that includes one or more of the following peaks two-theta (2θ): 5.6; 12.0; 14.0; 15.0; 15.5; 16.8; 18.1; 20.1; and 26.0. In the above XRPD pattern shown in Figure 57, Form B of 1:0.4 ponatinib / 2-methyl THF solvate exhibits at least one or all of the following peaks in degrees two-theta (2θ): 5.6; 12.0; 14.0; 20.1; 22.1; and 26.0. In certain embodiments, Form B of 1:0.4 ponatinib / 2-methyl THF solvate is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 5.6; 14.0; 20.1; and 26.0. In certain embodiments, the XRPD pattern of Form B of 1:0.4 ponatinib / 2-methyl THF solvate exhibits two peaks, three peaks, four peaks, or five peaks selected from the peaks above. In certain embodiments, Form B of 1:0.4 ponatinib / 2-methyl THF solvate is characterized by an XRPD pattern substantially similar to the top XRPD pattern in Figure 57. In certain embodiments, crystalline Form B comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees two-theta as set forth in the top pattern in Figure 57.

[0191] In the middle XRPD pattern shown in Figure 57, Form B 1:1 ponatinib / cyclohexanone solvate exhibits at least one or all of the following peaks at angles two theta (2θ): 5.6; 9.5; 11.0; 12.0; 12.9; 14.0; 15.2; 16.2; 16.8; 17.0; 18.1; 18.8; 19.5; 20.1; 21.8; 22.3; 23.0; 24.5; 26.1; 27.5; 28.0; and 28.2. In certain embodiments, Form B of the 1:1 ponatinib / cyclohexanone solvate is characterized by an XRPD pattern that includes one or more of the following peaks two-theta (2θ): 5.6; 11.0; 12.0; 12.9; 14.0; 15.2; 16.8; 18.1; 18.8; 20.1; 21.8; 22.3; 24.5; and 26.1. In the middle XRPD pattern shown in Figure 57, Form B of the 1:1 ponatinib / cyclohexanone solvate exhibits at least one or all of the following peaks in degrees two-theta (2θ): 5.6; 14.0; 15.2; 16.8; 20.1; 21.8; 22.3; and 26.1. In certain embodiments, Form B of the 1:1 ponatinib / cyclohexanone solvate is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 5.6; 14.0; 15.2; 16.8; 20.1; 22.3; and 26.1. In certain embodiments, the XRPD pattern of Form B of the 1:1 ponatinib / cyclohexanone solvate exhibits two, three, four, or five peaks selected from the peaks listed above. In certain embodiments, Form B of the 1:1 ponatinib / cyclohexanone solvate is characterized by an XRPD pattern substantially similar to the central XRPD pattern in Figure 57. In certain embodiments, crystalline Form B comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees two-theta as shown in the central pattern in Figure 57.

[0192] Figure 58 is a DSC curve obtained for B-Class 1:1 ponatinib / cyclohexanone solvate (QSA7.1) peak = 110.1°C and T peak An endothermic event is observed at 198.6°C.

[0193] Figure 59 is a plot showing the TGA and SDTA thermograms of B-Class 1:1 ponatinib / cyclohexanone solvate (QSA7.1).

[0194] TGMS data for the B-class 1:1 ponatinib / cyclohexanone solvate (QSA7.1) showed a mass loss of 14.8% (cyclohexanone and water) and 0.2% (cyclohexanone) occurring within the temperature intervals of 50-160 °C and 160-210 °C, respectively. The ponatinib / cyclohexanone ratio was estimated to be approximately 1.0 / 0.96 from the TGMS data.

[0195] An experiment was performed to characterize the purity of B-class 1:1 ponatinib / cyclohexanone solvate (QSA7.1) using HPLC. HPLC revealed a purity of 99.7460% (area percent) for B-class 1:1 ponatinib / cyclohexanone solvate (QSA7.1).

[0196] Figure 60 is a DSC curve obtained for B Class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1); peak = 67.1°C and T peak An endothermic event is observed at 197.5°C.

[0197] Figure 61 is a plot showing the TGA and SDTA thermograms of B Class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1).

[0198] TGMS data for the B-class 1:0.4 ponatinib / 2-methylTHF solvate (GEN8.1) showed mass losses of 3.1% (2-methylTHF), 1.9%, and 1.0% (2-methylTHF) occurring within the temperature intervals of 40–90 °C, 90–165 °C, and 165–215 °C, respectively. The ponatinib / 2-methylTHF ratio was estimated to be approximately 1.0 / 0.4 from the TGMS data.

[0199] An experiment was performed to characterize the purity of B-class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1) using HPLC. HPLC revealed a purity of B-class 1:0.4 ponatinib / 2-methyl THF solvate (GEN8.1) of 99.5939% (area percent).

[0200] Characteristics of Form C (poorly crystalline) ponatinib polymorph Poorly crystalline Form C can be obtained from crystalline Form A in the solvent methanol in a slurry experiment. Form C was found to contain solvated methanol at a ponatinib / methanol ratio of approximately 1:0.2.

[0201] Figure 62 shows the DSC curve obtained for the low-crystalline form C (GEN3.1), and T peak = 95.9 °C, endothermic event, T peak = 135.3°C and a pyrogenic event occurred. peak An endothermic event is observed at 198.1°C.

[0202] Figure 63 is a plot showing the TGA and SDTA thermograms of the less crystalline Form C (GEN3.1).

[0203] TGMS data for Form C (GEN3.1) showed a 1.3% mass loss (methanol) occurring within the temperature interval of 40-150° C. The ponatinib / methanol ratio was estimated to be approximately 1.0 / 0.2 from the TGMS data.

[0204] Form C was analyzed by X-ray powder diffraction (XRPD). Figure 64 shows the XRPD pattern of the overlaid XRPD of the starting material crystalline Form A (lower pattern) and the less crystalline Form C (upper pattern).

[0205] In the XRPD pattern shown in Figure 64, Form C exhibits at least one or all of the following peaks at angles 2-theta (2θ): 3.2; 11.1; 11.7; 12.8; 13.3; 13.5; 14.2; 17.1; 18.2; 20.8; 22.3; and 26.5. In certain embodiments, Form C is characterized by an XRPD pattern that includes one or more of the following peaks at angles 2-theta (2θ): 3.2; 12.8; 14.2; 17.1; 18.2; 20.8; 22.3; and 26.5. In the XRPD pattern shown in Figure 64, Form C exhibits at least one or all of the following peaks at angles 2-theta (2θ): 3.2; 12.8; 14.2; and 18.2. In certain embodiments, the XRPD pattern of Form C exhibits two, three, four, or five peaks selected from the peaks described above. In certain embodiments, Form C is characterized by an XRPD pattern substantially similar to the top XRPD pattern in Figure 64. In certain embodiments, crystalline Form C comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in Figure 64.

[0206] An experiment was performed to determine the purity of Form C (GEN3.1) using HPLC. HPLC revealed that Form C of ponatinib was 99.5725% (area percent) pure.

[0207] Characterization of the D-ponatinib polymorph Form D can be obtained from crystalline Form A by rapid anti-solvent crystallization in N,N-dimethylacetamide (DMA). Form D thus produced was found to be a DMA solvate with a ponatinib / DMA ratio of approximately 1:1.

[0208] Figure 65 shows the DSC curve obtained for Type D (GEN5.1R1), and T peak = 103.3℃, ​​T peak = 125.6°C and T peak An endothermic event is observed at 198.4°C.

[0209] FIG. 66 is a plot showing the TGA and SDTA thermograms of the poorly crystalline Form D (GEN5.1R1).

[0210] TGMS data for Form D (GEN5.1R1) showed a 13.8% mass loss (N,N-dimethylaceamide) occurring within the temperature interval of 40-140 °C. The ponatinib / DMA ratio was estimated to be approximately 1.0 / 0.98 from the TGMS data.

[0211] Form D was analyzed by X-ray powder diffraction (XRPD). Figure 67 shows the XRPD patterns of (bottom to top) the starting crystalline Form A; Form D (GEN5.1R1); and an overlaid XRPD of the rerun after 3 days (GEN5.1R4), indicating that the analyzed sample may contain Form D as well as the B class form.

[0212] In the central XRPD pattern shown in Figure 67, Form D exhibits at least one or all of the following peaks at angles two theta (2θ): 6.2; 8.0; 10.8; 11.5; 12.4; 13.5; 13.8; 14.5; 15.6; 16.5; 17.6; 18.5; 19.3; 19.8; 20.1; 20.8; 21.6; 22.1; 23.8; 26.0; 27.1; and 29.6. In certain embodiments, Form D is characterized by an XRPD pattern that includes one or more of the following peaks two theta (2θ): 6.2; 10.8; 12.4; 13.8; 14.5; 15.6; 16.5; 18.5; 20.2; 20.8; 21.6; 26.0; and 27.1. In the central XRPD pattern shown in Figure 67, Form D exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.2; 12.4; 14.5; 15.6; 16.5; 18.5; 20.2; 20.8; 21.6; 26.0; and 27.1. In certain embodiments, Form D is characterized by an XRPD pattern including one or more of the following peaks 2-theta (2θ): 6.2; 15.6; 16.5; 18.5; 20.2; 21.6; 26.0; and 27.1. In certain embodiments, the XRPD pattern of Form D exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form D ponatinib is characterized by an XRPD pattern substantially similar to the central XRPD pattern in Figure 67. In certain embodiments, crystalline Form D comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in FIG.

[0213] HPLC was used to perform a purity study of Form D. HPLC revealed that Form D ponatinib was 99.5056% (area percent) pure.

[0214] Characteristics of E-type (E-class) ponatinib polymorphism Class E solvents are prepared from either crystalline Form A or the less crystalline Form C in slurry and solubility experiments using solvents such as tetrahydrofuran (THF), chloroform, and dichloromethane (DCM), or by vapor diffusion onto the solid.

[0215] Based on thermal analysis, one representative sample of Form E was assigned to be a 1:1 ponatinib / tetrahydrofuran (THF) solvate. Another representative sample of Form E was likely a chloroform solvate, but this material was not further characterized other than by XRPD.

[0216] Figure 68 shows the DSC curve obtained for E-clasponatinib / THF 1:1 solvate (GEN7.1). peak = 95.9°C and T peak An endothermic event is observed at 198.1°C.

[0217] Figure 69 is a plot showing the TGA and SDTA thermograms of E-clasponatinib / THF 1:1 solvate (GEN7.1).

[0218] TGMS data for the E-class ponatinib / THF 1:1 solvate (GEN7.1) showed a mass loss of 11.7% (THF) occurring within the temperature interval of 40–130 °C. The ponatinib / THF ratio was estimated to be approximately 1.0 / 0.98 from the TGMS data.

[0219] The two Class E polymorphs were analyzed by X-ray powder diffraction (XRPD). Figure 70 shows the XRPD patterns of the overlaid XRPDs (from bottom to top) of the starting material crystalline Form A; the E-class ponatinib / THF 1:1 solvate (GEN7.1); and the E-class ponatinib / chloroform solvate (SLP3.1).

[0220] In the upper XRPD pattern shown in Figure 70, E-clasponatinib / THF 1:1 solvate (GEN7.1) exhibits at least one or all of the following peaks at angles two-theta (2θ): 6.2; 7.0; 10.0; 13.0; 15.1; 16.4; 17.2; 18.5; 20.4; 20.5; 22.5; 24.4; 25.6; and 27.0. In certain embodiments, E-clasponatinib / THF 1:1 solvate (GEN7.1) is characterized by an XRPD pattern that includes one or more of the following peaks at angles two-theta (2θ): 6.2; 7.0; 10.0; 15.1; 16.4; 17.2; 18.5; 20.4; 20.5 24.4; and 27.0. In the XRPD pattern shown in Figure 70, E-classponatinib / THF 1:1 solvate (GEN7.1) exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.2; 15.1; 16.4; 17.2; 18.5; 20.4; 24.4; and 27.0. In certain embodiments, E-classponatinib / THF 1:1 solvate (GEN7.1) is characterized by an XRPD pattern that includes one or more of the following peaks at angles 2-theta (2θ): 6.2; 15.1; 16.4; 20.4; and 20.5. In certain embodiments, the XRPD pattern of E-classponatinib THF 1:1 solvate (GEN7.1) exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, E-clasponatinib / THF 1:1 solvate is characterized by an XRPD pattern substantially similar to the top XRPD pattern in Figure 70. In certain embodiments, crystalline Form E comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in the top pattern in Figure 70.

[0221] In the middle XRPD pattern shown in Figure 70, E-clasponatinib / chloroform solvate (SLP3.1) exhibits at least one or all of the following peaks at angles two theta (2θ): 6.2; 7.0; 8.7; 9.8; 12.1; 12.5; 13.0; 15.2; 16.4; 17.2; 18.5; 20.0; 21.0; 23.0; 24.4; 25.0; and 26.2. In certain embodiments, E-classponatinib / chloroform solvate (SLP3.1) is characterized by an XRPD pattern comprising one or more of the following peaks 2-theta (2θ): 6.2; 7.0; 13.0; 15.2; 16.4; 17.2; 18.5; 20.0; 21.0; 24.4; 25.0; and 26.2. In certain embodiments, the XRPD pattern of E-classponatinib / chloroform solvate (SLP3.1) exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, E-classponatinib / chloroform solvate is characterized by an XRPD pattern substantially similar to the central XRPD pattern in Figure 70. In certain embodiments, crystalline Form E comprises a powder X-ray diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in the central pattern in Figure 70.

[0222] An experiment was performed to determine the purity of the E-class ponatinib / THF 1:1 solvate (GEN7.1) using HPLC. HPLC revealed that the purity of ponatinib E-class ponatinib THF 1:1 solvate (GEN7.1) was 99.5120% (area percent).

[0223] Characteristics of the F-form ponatinib polymorph Form F was formed from Form A under certain experimental conditions, such as in the presence of a polar solvent. Analysis of Form F by TGMS revealed that Form F initially loses 3.3% of water in the temperature range 25-140 °C. From these results, it could be inferred that the first endothermic event observed corresponds to a dehydration process and that Form F is a hydrated form, e.g., a 1:1 hydrate with water.

[0224] From the screening experiment, selected samples of pure Form F (6 samples) and mixtures of Forms A and F (6 samples) were stored at ambient temperature for 4 months and then re-analyzed by XRPD, the results of which are shown in Table 24 below.

[0225] [Table 26]

[0226] The results in Table 24 show that the polymorphism remains unchanged in most cases. Results from anti-solvent rapid crystallization experiments (2-methoxyethanol / water) showed that the resulting form F material converted to form A after 4 months if the sample was collected wet. Anti-solvent experiments using acetone / water also showed that form F converted to a mixture of forms A and F after 4 months.

[0227] Figure 71 shows the TGA and SDTA plots obtained for Form F (AS16.2). Form F (AS16.2) exhibited an initial endotherm followed by recrystallization in the temperature range of 130-140°C. A melting point of approximately 189°C was observed for Form F (AS16.2).

[0228] As noted above, the TGMS data for Form F (AS16.2) showed a 3.3% mass loss (water) occurring within the temperature interval of 25 to 140° C. The ponatinib / water ratio of Form F was estimated from the TGMS data to be approximately 1.0 / 1.01.

[0229] Form F (SLP10.1) was analyzed by X-ray powder diffraction (XRPD). Figure 83 shows a stack of XRDP patterns, with Form F (SLP10.1) being the fifth pattern from the top. This sample (SLP10.1) was obtained in a slurry experiment using 1,2-dimethoxyethane as the solvent.

[0230] In the XRPD pattern shown as the fifth pattern from the top in Figure 83, Form F of ponatinib exhibits at least one or all of the following peaks at angles two-theta (2θ): 7.2; 13.2; 14.1; 15.9; 18.1; 20.4; 21.1; 22.0; 23.5; 24.2; 25.5; and 26.8. In certain embodiments, Form F is characterized by an XRPD pattern that includes one or more of the following peaks at angles two-theta (2θ): 7.2; 13.2; 14.1; 15.9; 18.1; 20.4; 23.5; 25.5; and 26.8. In the XRPD pattern shown as the fifth pattern from the top in Figure 83, Form F exhibits at least one or all of the following peaks in degrees two-theta (2θ): 7.2; 14.1; 18.1; 20.4; 25.5; and 26.8. In certain embodiments, the XRPD pattern of Form F exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form F of ponatinib is characterized by an XRPD pattern substantially similar to the fifth XRPD pattern from the top in Figure 83. In certain embodiments, crystalline Form F comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees two-theta as shown in the fifth pattern from the top in Figure 83.

[0231] Characteristics of H-type (H-class) ponatinib polymorphism Characteristics of ponatinib / 1-propanol 1:1 solvate form H Dissolution experiments with 1-propanol revealed that the recovered solid was the 1-propanol solvate of Form H.

[0232] The DSC curve obtained for H-clasponatinib / 1-propanol solvate (SAS35) shows a T of approximately 93°C. peak and a T of approximately 192°C peak The endothermic event occurring at

[0233] TGMS data for the H-class ponatinib / 1-propanol solvate (SAS35) showed a 9.2% mass loss (1-propanol) occurring within the temperature interval of 25–120 °C. The ponatinib / 1-propanol ratio was estimated to be approximately 1.0 / 0.9 for the H-class ponatinib / 1-propanol solvate (SAS35) from the TGMS data. Another sample (SAS30) showed a mass loss corresponding to a 1:0.6 solvate, but it was later determined that this sample converted partially back to the crystalline anhydrous Form A, thereby reducing the overall solvent loss observed in the bulk sample.

[0234] The H-class solvate (SAS35) was analyzed by X-ray powder diffraction (XRPD) and exhibited at least one or all of the following peaks at angles two-theta (2θ): 6.1; 6.8; 10.0; 12.0; 13.2; 13.5; 16.0; 16.5; 16.0; 16.5; 18.0; 19.0; 19.5; 20.4; 21.0; 22.5; 25.0; 25.5; 26.2; 27.0; and 27.5. In certain embodiments, Form H is characterized by an XRPD pattern that includes one or more of the following peaks two-theta (2θ): 12.0; 13.2; 13.5; 18.0; 25.0; and 25.5. In certain embodiments, the XRPD pattern of Form H exhibits two, three, four, or five peaks selected from the peaks described above.

[0235] Characteristics of the ponatinib / 2-methoxyethanol 1:1 solvate form H Ponatinib free base can be formed from isomorphous solvates of Form H in other alcohols. Starting with Form A as the starting material, the 2-methoxyethanol solvate was obtained by vapor diffusion into liquid using 2-methoxyethanol as the solvent. The purity of this form was estimated to be 98.0%. TGMS confirmed this to be a 1:0.93 ponatinib / 2-methoxyethanol solvate, which desolvated at approximately 96°C. After desolvation, a melting point of approximately 198.8°C was observed by DSC, which corresponds approximately to the melting point of Form A.

[0236] The DVS experiment showed a weight loss of approximately 3.6% upon desorption, which was not regained upon sorption at 45% RH. XRPD analysis at the end of the DVS experiment showed no physical changes. Partial desolvation may have occurred, but this process did not cause a collapse of the crystalline structure (as evidenced by the approximately 12% weight loss observed by TG-MS upon desolvation).

[0237] However, Form H converted to Form A after 10 months of storage under ambient conditions as well as after 1 week of accelerated stress conditions (40°C, 75% RH) in a humidity chamber.

[0238] Figure 72 shows the DSC curve obtained for Form H (VLD1, dry solid from stock), and T peak = 96.1 °C (desolvation event) and T peak = 198.8°C. The endothermic event at 198.8°C is likely the melting event for Form A.

[0239] Figure 73 is an overlay of characteristic TGA and SDTA thermograms of Form H (VLD1, dried solid from stock).

[0240] DVS experiments on Form H (VLD1, dried solid from stock) showed a weight loss (2-methoxyethanol) of approximately 11.7% in the temperature range of 25–120 °C. These results indicate that the first endothermic event observed corresponds to the desolvation process, and that Form H (VLD1, dried solid from stock) is a 1:0.93 ponatinib / 2-methoxyethanol solvate.

[0241] Figure 74 shows a series of XRPD patterns obtained for various samples of Form H ponatinib, along with the XRPD of Form A. In Figure 74: Plot 1 is Form H (VLD2 experiment, after 2 weeks and drying); Plot 2 is Form H (VLD1 experiment, after 2 weeks and drying); Plot 3 is Form H (VLD1 experiment, dried solid from stock after DVS); Plot 4 is Form H (VLD1, dried solid from stock); Plot 5 is Form H (VLD19); and the bottom plot is the XRPD of Form A. The XRPD patterns shown as Plots 1-5 are substantially identical.

[0242] In the XRPD pattern shown as plot 5 in Figure 74, Form H of ponatinib exhibits at least one or all of the following peaks at angles two-theta (2θ): 6.2; 6.5; 10.1; 12.0; 13.2; 15.0; 15.5; 16.0; 16.5; 18.0; 19.1; 19.6; 20.5; 21.1; 23.0; 23.7; and 25.5. In certain embodiments, Form H is characterized by an XRPD pattern including one or more of the following peaks at angles two-theta (2θ): 6.2; 12.0; 13.2; 16.0; 18.0; 19.1; 19.6; 20.5; 21.1; 23.0; and 25.5. In the XRPD pattern shown as plot 5 in Figure 74, Form H exhibits at least one or all of the following peaks at angles two-theta (2θ): 6.2; 12.0; 18.0; and 25.5. In certain embodiments, the XRPD pattern of Form H exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form H ponatinib is characterized by an XRPD pattern substantially similar to any one of patterns 1-5 in Figure 74. In certain embodiments, crystalline Form H comprises an X-ray powder diffraction pattern having characteristic peaks expressed in angles two-theta shown in any one of patterns 1-5 in Figure 74.

[0243] An experiment was performed to determine the purity of Form H (VLD1 dried solid from stock) using HPLC. HPLC revealed that the purity of Form H ponatinib (VLD1 dried solid from stock) was 98.0464% (area percent).

[0244] Figure 75 shows an overlay of the characteristic FT-IR spectrum obtained from Form H of ponatinib (VLD1 dried from stock) (Plot 1) and Form A (Plot 2). The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm). -1 ) is shown.

[0245] Figure 76 shows an overlay of the characteristic FT-IR spectrum (plot 1) obtained from Form H of ponatinib (VLD1 dried from stock) and the FT-IR spectrum (plot 2) obtained from Form A in the wavelength region of 1750-600 nm. The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm). -1 ) is shown. In this superposition, several peaks characteristic of Form H as well as several peaks characteristic of Form A are observed. In this regard, some peaks characteristic of Form A (plot 2) in Figure 76 are: 1605 cm -1 ;1415cm -1 ;1295cm -1 ;1250cm -1 ;1150cm -1 ;1145cm -1 ;1110cm -1 ;1100cm -1 ;895cm -1 ;855 -1 ; and 790cm -1 Examples include:

[0246] Characteristics of Form I Ponatinib Low-Crystalline Polymorph The poorly crystalline Form I was obtained by freeze-drying technique in dichloromethane (DCM).

[0247] Based on thermal analysis, Form I (GEN9.1) contains a small level of solvated DCM, corresponding to a ponatinib / DCM ratio of approximately 1:0.03.

[0248] Figure 77 shows the DSC curve obtained for Type I (GEN9.1), and T peak An exothermic event occurred at T = 100.1°C. peak An endothermic event is observed at 196.7°C.

[0249] Figure 78 is a plot showing the TGA and SDTA thermograms of Form I (GEN9.1).

[0250] TGMS data for Form I (GEN9.1) showed a 0.5% mass loss (DCM) occurring within the temperature interval of 40-175°C (it was slightly solvated). The ponatinib / DCM ratio was estimated to be approximately 1.0 / 0.03 from the TGMS data.

[0251] Form I (GEN9.1) was analyzed by X-ray powder diffraction (XRPD). Figure 77 shows (from bottom to top) an XRPD overlay of starting material crystalline Form A and Form I (GEN9.1).

[0252] In the upper XRPD pattern shown in Figure 79, Form I (GEN9.1) exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.5; 8.2; 9.8; 14.3; 15.5; 17.5; 21.2; 23.1; and 26.5. In certain embodiments, Form I (GEN9.1) is characterized by an XRPD pattern that includes one or more of the following peaks at angles 2-theta (2θ): 6.5; 8.2; 9.8; 14.3; 15.5; 17.5; 21.2; and 26.5. In the upper XRPD pattern shown in Figure 79, Form I (GEN9.1) exhibits at least one or all of the following peaks at angles 2-theta (2θ): 6.5; 8.2; 15.5; 17.5; 21.2; and 26.5. In certain embodiments, Form I (GEN9.1) is characterized by an XRPD pattern comprising one or more of the following peaks 2-theta (2θ): 8.2; and 15.5. In certain embodiments, the XRPD pattern of Form I (GEN9.1) exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form I ponatinib is characterized by an XRPD pattern substantially similar to the top pattern in Figure 79. In certain embodiments, crystalline Form I comprises a powder X-ray diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in the top pattern in Figure 79.

[0253] HPLC was used to perform a purity study of Form I (GEN9.1). HPLC revealed that Form I (GEN9.1) ponatinib was 99.5802% (area percent) pure.

[0254] Characteristics of the J-type ponatinib polymorph Form J was obtained by vapor diffusion onto solid. Purity was estimated to be 96.5%. TGMS analysis (11.4% weight loss of thiophene between 25 and 130 °C) confirmed that Form J "poorly crystalline" is a 1:0.82 ponatinib / thiophene solvate that desolvates at 83.7 °C. Following desolvation, a melting point event occurred at approximately 197.3 °C, which is likely the melting of Form A.

[0255] The DVS experiment showed a weight loss of approximately 1.3% during desorption. The sorption phase did not appear to regain weight (mass uptake approximately 0.33%) until at least 45% relative humidity (RH). XRPD analysis at the end of the DVS experiment showed that the material converted to Form A.

[0256] The poorly crystalline form J also converted to form A after 10 months of storage under ambient conditions and after 1 week under stress conditions (40°C, 75% RH).

[0257] Two samples of Form J of ponatinib (designated VDS2 and VDS10 for Screen S10010A) were analyzed by powder X-ray diffraction (XRPD). Figure 78 shows an overlay of XRPD patterns (from top to bottom): Plot 6 is the XRPD pattern of Form A (VDS2, after stability testing); Plot 7 is the XRPD pattern of Form J (VDS2); Plot 8 is the XRPD pattern of Form J (VDS10 in Screen S10010A); and Plot 9 is another XRPD pattern of Form A of ponatinib.

[0258] In XRPD plots 7 and 8 of Figure 80, Form J of ponatinib exhibits at least one or all of the following peaks at angles two-theta (2θ): 5.8; 7.0; 12.1; 15.1; 16.8; 18.1; 18.6; 19.1; 19.5; 20.1; 21.1; 21.8; 22.8; 25.0; 25.7; and 27.0. In certain embodiments, Form J of ponatinib is characterized by an XRPD pattern including one or more of the following peaks at angles two-theta (2θ): 5.8; 7.0; 12.1; 15.1; 16.8; 18.1; 19.1; 19.5; 20.1; 21.1; 21.8; 22.8; 25.0; 25.7; and 27.0. In the XRPD pattern of Figure 80 above, Form J of ponatinib exhibits at least one or all of the following peaks at angles 2-theta (2θ): 5.8; 7.0; 12.1; 15.1; 16.8; 18.6; 19.1; 19.5; 21.8; 22.8; 25.0; 25.7; and 27.0. In certain embodiments, Form J of ponatinib is characterized by an XRPD pattern comprising one or more of the following peaks at angles 2-theta (2θ): 5.8; 7.0; 12.1; 15.1; 18.6; 19.5; 21.8; 25.0; 25.7; and 27.0. In certain embodiments, the XRPD pattern of Form J of ponatinib exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form J of ponatinib is characterized by an XRPD pattern substantially similar to either one of plots 7 and 8 of Figure 80. In certain embodiments, crystalline Form J comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as set forth in either one of plots 7 and 8 of Figure 80.

[0259] Purity experiments for Form J (VDS2) were performed using HPLC, which revealed that Form J (VDS2) ponatinib was 96.4509% (area percent).

[0260] Figure 81 shows a characteristic FT-IR spectrum obtained from Form J (VDS2) ponatinib (Plot 1) overlaid with the FT-IR spectrum obtained from Form A (Plot 2). The vertical axis shows percent transmittance (%) and the horizontal axis shows wavenumber (cm). -1 ) is shown.

[0261] Figure 82 shows a superposition of the characteristic FT-IR spectrum (plot 1) obtained from Form J (VDS2) ponatinib and the FT-IR spectrum (plot 2) obtained from Form A in the wavelength region of 1750 to 600 nm. The vertical axis shows percent transmittance (%), and the horizontal axis shows wavenumber (cm -1 ) is shown.

[0262] Characteristics of the K-type ponatinib polymorph Solid form K was found in sample SAS58 (MSZW experiment) at 25 mg / mL in 1-propanol / acetonitrile (30 / 70). The TGMS thermogram showed a very slight mass loss (less than 0.04% in the temperature interval 25-175 °C) before melting. The SDTA signal indicates a melting point of form K of 184 °C.

[0263] The DSC curve obtained for form K of ponatinib shows a T of approximately 184°C. peak The endothermic event occurring at

[0264] Form K of ponatinib has been analyzed by X-ray powder diffraction (XRPD) and displays at least one or all of the following peaks at angles two-theta (2θ): 10.0; 11.0; 13.4; 14.6; 15.2; 16.0; 17.0; 17.5; 18.0; 19.6; 20.9; 22.1; 22.8; 24.1; 24.8; 26.5; 27.1; 28.5; and 30.5. In certain embodiments, Form K is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 10.0; 11.0; 13.4; 14.6; 15.2; 16.0; 19.6; 20.9; 22.1; 22.8; 24.1; 24.8; and 26.5. In certain embodiments, Form K is characterized by an XRPD pattern comprising one or more of the following peaks two-theta (2θ): 10.0; 11.0; 13.4; 14.6; 15.2; 19.6; 22.1; 22.8; and 24.1. In certain embodiments, the XRPD pattern of Form K exhibits two, three, four, or five peaks selected from the above peaks.

[0265] Overlay of XRPD patterns of selected ponatinib polymorphs Figure 83 is an overlay of XRPD results obtained for selected ponatinib free base polymorphs. The XRPD patterns shown, from bottom to top: Form A (SM); B-class (QSAS7.1); Form C, poorly crystalline (GEN3.1); Form D (GEN5.1); E-class (SLP3.1); Form F (SLP10.1); Form G (AS19.1), described later herein; Form H (VDL19.1); Form I, poorly crystalline (GEN9.1); and Form J, poorly crystalline (VDS10.1).

[0266] Characteristics of the G-type ponatinib polymorph Figure 83 illustrates Form G of ponatinib obtained by a rapid crystallization / anti-solvent process in which the solvent used was 3-methyl-1-butanol and the anti-solvent used was cyclohexane. Form G of ponatinib was analyzed by X-ray powder diffraction (XRPD) and exhibited at least one or all of the following peaks at angles two theta (2θ): 5.0; 6.5; 9.5; 12.0; 12.5; 14.0; 15.0; 16.5; 17.2; 18.4; 20.0; 21.0; 22.8; 23.5; 24.6; and 29.5. In certain embodiments, Form G is characterized by an XRPD pattern comprising one or more of the following peaks 2-theta (2θ): 5.0; 6.5; 9.5; 14.0; 15.0; 16.5; 17.2; 18.4; 20.0; and 22.8. In certain embodiments, Form G is characterized by an XRPD pattern comprising one or more of the following peaks 2-theta (2θ): 5.0; 6.5; 9.5; 14.0; 17.2; 18.4; 20.0; and 22.8. In certain embodiments, the XRPD pattern of Form G exhibits two, three, four, or five peaks selected from the above peaks. In certain embodiments, Form G of ponatinib is characterized by an XRPD pattern substantially similar to the fourth pattern from the top of the overlaid patterns in Figure 83. In certain embodiments, crystalline Form G comprises an X-ray powder diffraction pattern having characteristic peaks expressed in degrees 2-theta as shown in the fourth pattern from the top in Figure 83.

[0267] Example 6 Discovery of ponatinib polymorphs Initial efforts to discover polymorphs of ponatinib were divided into two phases. Phase 3 included characterization of the starting material, feasibility studies, and solubility studies to provide data for solvent selection in Phase 4. Phase 4 included 192 milliliter (ml)-scale polymorph screening experiments. These initial efforts led to the discovery of 11 polymorphs of ponatinib free base: Forms A, B, C, D, E, F, G, H, I, J, and K.

[0268] Phase 1: Characterization of starting materials The synthesis described later herein yielded the compound ponatinib free base, which was characterized by the amide coupling reaction of the amine subunit with the methyl ester subunit. Approximately 20 grams of the free base (total of two batches designated F09-05575 and F09-05576) was obtained as a pale yellow solid. The starting material was characterized by XRPD, digital imaging, DSC, TGMS, and HPLC.

[0269] Figure 84 shows the XRPD patterns of the free base starting material, with two patterns representing the two batches mentioned above (F09-05575: lower pattern; and F09-05576: upper pattern).

[0270] Figure 85 is the DSC curve for ponatinib free base starting material batch F09-05575; peak = 182.6°C and T peak = 199.0°C (major). The major endotherm corresponds to a melting event and is likely accompanied by a desolvation process (observed in the TGA / SDTA trace of this batch).

[0271] Figure 86 is the DSC curve for ponatinib free base starting material batch F09-05576; peak = 199.6°C. The endotherm corresponds to a melting event and is likely accompanied by a desolvation process (observed in the TGA / SDTA trace of this batch).

[0272] Figure 87 is a plot showing the TGA and SDTA thermograms of ponatinib free base starting material batch F09-05575.

[0273] Figure 88 is a plot showing the TGA and SDTA thermograms of ponatinib free base starting material batch F09-05576.

[0274] TGA and TGMS analyses showed that batch F09-05575 experienced only a single mass loss (0.9% over the temperature range of 25-210°C), while batch F09-05576 experienced two mass losses (0.3% over the temperature range of 25-120°C and 0.9% over the temperature range of 120-210°C). The mass losses observed in both batches correspond to 2-methyltetrahydrofuran (confirmed by the m / z ratios observed in the MS data). The presence of this solvent in residual amounts is believed to be from the final synthesis step of the ponatinib free base compound.

[0275] Purity experiments were performed on ponatinib free base compound using HPLC, which revealed a purity of 99.5166% (area percent) for batch F09-05575 and 99.6869% (area percent) for batch F09-05576.

[0276] In various embodiments, alternative methods for preparing Form A resulted in samples of varying crystallinity.

[0277] Alternative Methods for Preparing Form A of Ponatinib Free Base Preparation 1 A 180 g quantity of M010578 sample of ABL411057 (2-Me-THF, 1.1 equivalents of aniline, 1.6 equivalents of KOtBu) was crystallized from neat 1-propanol, and the 1-PrOH wet product was then triturated in neat acetonitrile to give Form A with a purity of 99.39 a%.

[0278] Preparation 2 Two 180 g samples of ABL411060 (2-Me-THF, 26 °C IT, 1.1 equiv. aniline, 1.6 equiv. KOtBu) were obtained from the 1-PrOH solution of M010578 obtained after solvent exchange from 2-Me-THF to 1-PrOH. The 1-PrOH solution was split 9:1.

[0279] 779.5 g (9 portions) of the above solution of M010578 in 1-PrOH was crystallized overnight at ambient temperature. After filtration, the wet cake was triturated in 160 g of acetonitrile at 40° C., filtered, and dried (50° C., 3 mbar) to yield 211.6 g (99.87 a%) of Form A of ponatinib free base.

[0280] To 86.6 g (1 portion) of the above solution of M010578 in 1-PrOH was added 260 g of acetonitrile. After 1 hour, the suspension was filtered, and the filter cake was washed with ACN / 1-PrOH (3:1 v / v) and dried to give 25.6 g (99.27 a%) of the free base. Crystallization and isolation from 1-propanol, followed by trituration in acetonitrile, afforded a product of higher HPLC purity than isolation by precipitation from the ACN / 1-PrOH mixture.

[0281] Stage 1: Solubility test A quantitative solubility study of the ponatinib free base starting material was conducted using a set of 20 solvents. Slurries were prepared with an equilibration time of 24 hours, and then the slurries were filtered. Solubility was determined from saturated solutions by HPLC. Residual solids were characterized by XRPD. The results are summarized in Table 25 below.

[0282] [Table 27]

[0283] The material obtained from 19 of the 22 different solubility evaluations was the same polymorph as the starting free base, designated Form A. The solid obtained from the cyclohexanone slurry exhibited a different XRPD and was designated Class B Form, which was further characterized as four solvates as described above.

[0284] Feasibility study of the ponatinib free base compound A feasibility study was conducted to attempt to obtain amorphous free base material that could be used in several Stage 4 crystallization techniques. Two techniques were used: milling and freeze-drying. The results are shown below.

[0285] crushing Two milling experiments were carried out at a frequency of 30 Hz for two different durations, as summarized below in Table 26. After 60 or 120 minutes of milling, the material remained crystalline (Form A).

[0286] [Table 28]

[0287] Freeze drying Eight lyophilization experiments were performed with the ponatinib free base compound and are summarized in Table 27 below.

[0288] [Table 29]

[0289] Solvates such as 2,2,2-trifluoroethanol (TFE) and TFE / water mixtures yielded amorphous material. In experiments conducted with 2-methyltetrahydrofuran, DMA / water (90:10), and THF, three novel crystalline forms were observed, designated B-class, D-class, and E-class. In the fourth stage of screening, additional B-class and E-class forms were observed, which the results suggest are isomorphous. Remaining experiments conducted with methanol and dichloromethane yielded two low-crystalline materials (Form C low-crystalline and Form I low-crystalline, respectively).

[0290] The novel crystalline forms were further analyzed and characterized by DSC, TGMS, and HPLC, and Forms D and E were confirmed to be solvates (1:1 API / DMA and 1:1 API / THF, respectively).

[0291] The low-crystalline material obtained from methanol (C low-crystalline) and dichloromethane (I low-crystalline) had low amounts of residual solvent (1.3% in the temperature range 40°C to 150°C and 0.5% in the temperature range 40°C to 175°C, respectively).

[0292] Because the freeze-drying method using methanol was not optimal for producing poorly crystalline material, the dichloromethane method was chosen to produce this material for use in the fourth stage cooling / evaporative crystallization and vapor diffusion onto solid experiments.

[0293] Solvents for Phase 4 experiments were selected based on the results of the feasibility study and dissolution behavior of ponatinib free base.

[0294] Solvent Evaluation To select screening solvents and determine the concentration ranges used in the screening, quantitative solubility studies were performed on batch F09-05575 of the free base starting material. A set of 20 solvents was used for this screening. For each solvent, a standard 1.8 ml screw-cap vial was charged with 40 mg of starting material, 200 μl of solvent, and a magnetic stir bar. The vial was then capped and equilibrated at 25°C for 24 hours with stirring. The resulting mixture (slurry) was filtered (0.5 microns), and the isolated mother liquor was diluted to two dilutions selected according to a calibration curve. The amount of API in the diluted solution was determined by HPLC analysis (DAD). Calibration curves were obtained from stock solutions of the two free base compounds prepared separately in 2,2,2-trifluoroethanol.

[0295] After the solubility was determined, residual solvent was evaporated from each vial (slurry) under vacuum at ambient temperature, and all resulting residues were analyzed by powder X-ray diffraction to determine the presence of novel crystalline forms.

[0296] Feasibility testing The experimental conditions for the feasibility study of the free base compounds are summarized below in Table 28. After the experiment, HPLC analysis was performed to determine purity and thermal analysis was performed to determine the thermal behavior of each form.

[0297] [Table 30]

[0298] Polymorphism screening experimental design and protocol Polymorph screening experiments for ponatinib free base compound were conducted at the milliliter (ml) scale using 192 different conditions and six different crystallization methods: cooling-evaporation, antisolvent addition, grinding, slurry, vapor diffusion into solution, and vapor diffusion onto solid.

[0299] Cooling-evaporation crystallization experiment Twenty-four ml-scale cryo-evaporation experiments were performed in 8 ml vials using 24 different solvents and one concentration. In each vial, 25 mg of ponatinib free base was added to a liquid (dichloromethane). The sample was lyophilized to obtain a powdery, poorly crystalline material. The screening solvent was then added until a concentration of approximately 60 mg / ml was reached (see Table 29 below). The vials were capped and subjected to the temperature profile described in Table 30 below. The mixtures were cooled to 5°C and held at that temperature for 48 hours, after which the vials were placed under vacuum. The solvent was evaporated at 200 mbar or 10 mbar for several days and analyzed by XRPD and digital imaging.

[0300] [Table 31]

[0301] [Table 32]

[0302] Rapid crystallization by adding antisolvent Forty-eight different crystallization conditions were used for the rapid crystallization experiments, using 23 different solvents and 18 different anti-solvents (see Table 31 below). For each solvent, a stock solution was prepared and, after 17 hours of equilibration, the concentration of ponatinib free base in each case reached its saturated concentration at ambient temperature and was then filtered into a set of 8 ml vials. To each of these vials, a different anti-solvent was added using a solvent to anti-solvent ratio of 1:0.25. If no precipitation occurred, this ratio was increased to 1:4 with a 60-minute wait time between additions. Solids that precipitated during the wait time between anti-solvent additions were isolated by centrifugation. If no solids were obtained, the solvent was completely evaporated under vacuum at room temperature. If solids were obtained, they were analyzed by XRPD and digital imaging.

[0303] [Table 33]

[0304] Crushing experiment In the droplet milling method, a small amount of solvent is added to the ponatinib free base raw material, which is then milled in a stainless steel milling jar containing two stainless steel milling balls. In this manner, the effect of 24 different solvents (see Table 32) was investigated. Typically, 30 mg of starting material was milled and analyzed.

[0305] [Table 34]

[0306] Slurry experiments A total of 48 slurry experiments were conducted using ponatinib free base and 24 different solvents at 10°C and 30°C, respectively, over a two-week period. Table 33 below summarizes the experimental conditions. The experiments were conducted by stirring the suspension of material in the solvent at a controlled temperature. At the end of the slurry period, the vials were centrifuged to separate the solid from the mother liquor. The solid was further dried under full vacuum at room temperature and analyzed by XRPD and digital imaging.

[0307] [Table 35]

[0308] Vapor diffusion into solution In vapor diffusion experiments, saturated solutions of ponatinib free base were exposed to solvent vapors at room temperature for two weeks. A volume of the saturated solution was transferred to an 8 ml vial, left open, and placed in a closed 40 ml vial containing 2 ml of antisolvent (see Table 34 below). After two weeks, the samples were checked for solid formation. If solids were present, the liquid was separated from the solids. Samples were dried under vacuum (200 mbar or 10 mbar) and then analyzed by XRPD and digital imaging.

[0309] [Table 36]

[0310] Vapor diffusion onto solids In this vapor diffusion experiment, amorphous ponatinib free base was exposed to solvent vapors at room temperature for two weeks. The API was added as a liquid to an 8 ml vial and then lyophilized. The 8 ml vial containing the amorphous material was left open and placed in a closed 40 ml vial containing 2 ml of antisolvent (see Table 35 below). After two weeks, the solid was analyzed by XRPD and digital imaging. If the solid had liquefied due to vapor, the sample was dried under vacuum (200 mbar or 10 mbar) before being analyzed by XRPD and digital imaging.

[0311] [Table 37]

[0312] Physical Stability and Scale-Up of Selected Ponatinib Polymorphs The purpose of this study was to reproduce and further characterize the solid-state forms of ponatinib identified in the studies described herein above. This study revealed that Form D (isomorphous solvate) and Form F (monohydrate) were physically stable for at least 10 months under ambient conditions. Forms B and E, as well as Forms G, H, I (poorly crystalline), and J (poorly crystalline), converted to Form A over the course of 10 months under ambient conditions.

[0313] Successful scale-up of Forms H and J (low crystallinity) was achieved. Attempted scale-up of Form G resulted in Form A. The scale-up project was carried out in three stages: Phase 1: The physical stability of the various forms obtained in previous studies will be examined by XRPD after storage under ambient conditions for 8-10 months; Phase 2: Scale-up to 50-120 mg to further characterize selected solid-state forms of ponatinib free base; and Step 3: Clarify the solvation state, thermal properties, and physical stability of the substance obtained in Step 2.

[0314] Phase 1 results Form D (isomorphous solvate) and Form F (monohydrate) are stable over the study period. Isomorphous solvates of Form B and Form E classes all converted to Form A. Forms G, H, I (poorly crystalline), and J (poorly crystalline) all converted to Form A. Figure 89 tabulates the physical stability of several solid forms of ponatinib.

[0315] Phase 2: Scale-up of selected ponatinib free base forms Forms G, H, and J (low crystallinity) were selected for scale-up studies. Scale-up experimental conditions were adapted from the polymorph screening disclosed herein. Scale-up was successful for Forms H and J (low crystallinity). Figure 90 tabulates the results of scale-up experiments for selected free base forms.

[0316] Phase 3: Characterization of the scaled-up form The solid forms previously scaled up and confirmed by XRPD were further characterized by DSC, TGMS, FTIR, HPLC, and DVS. Form A, which resulted from the attempted scale-up of Form G, was not further characterized. Additionally, the physical stability to accelerated aging conditions (40°C, 75% RH for 1 week) was investigated. Figure 91 tabulates the various characterizations of the ponatinib free base forms successfully reproduced at the 120 mg scale.

[0317] Pharmaceutical compositions and treatment of physiological conditions therewith The present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of ponatinib hydrochloride disclosed herein and at least one pharmaceutically acceptable carrier, excipient, or vehicle. In certain embodiments, a unit dosage form of the pharmaceutical composition comprises a single crystalline form of ponatinib hydrochloride as the API. Alternatively, a unit dosage form of the pharmaceutical composition comprises two or more crystalline forms of ponatinib hydrochloride. In certain embodiments, more than about 50%, more than about 70%, more than about 80%, or more than about 90% of the single crystalline form present in the composition is one of the selected forms. In any of the above embodiments, one or all of the crystalline forms are substantially pure. For example, in certain embodiments, a pharmaceutical composition comprises substantially pure Form A of ponatinib hydrochloride and at least one pharmaceutically acceptable carrier, excipient, or vehicle. Alternatively, a pharmaceutical composition comprises Form A and Form J of ponatinib hydrochloride and at least one pharmaceutically acceptable carrier, excipient, or vehicle. Other variations on this subject matter will be readily apparent to those skilled in the art having the benefit of this disclosure.

[0318] At least one pharmaceutically acceptable carrier, diluent, excipient, or vehicle can be easily selected by those skilled in the art and is determined by the desired method of administration. Specific examples of suitable administration methods include oral administration, nasal administration, parenteral administration, topical administration, transdermal administration, and rectal administration. The pharmaceutical compositions disclosed herein can take any pharmaceutical form recognized as appropriate by those skilled in the art. Suitable pharmaceutical forms include solid, semi-solid, liquid, or lyophilized formulations, such as tablets, powders, capsules, suppositories, suspensions, liposomes, and aerosols.

[0319] Various solid forms of ponatinib and various solid forms of ponatinib hydrochloride may be administered to a subject in need of treatment in a therapeutically effective amount, either alone or in any combination. Similarly, any solid forms of ponatinib and ponatinib hydrochloride disclosed herein may be formulated into pharmaceutical compositions, either alone or in any combination, which can then be used to treat various disease states in animals, including humans. For example, pharmaceutical compositions containing any single or combination of polymorphs of ponatinib and / or ponatinib hydrochloride may be used to treat CML or Ph+ALL in a subject in need thereof by administering a therapeutically effective amount of the pharmaceutical composition to the subject in need thereof.

[0320] III. Synthesis of Ponatinib and Ponatinib Hydrochloride Ponatinib free base and ponatinib HCl are the products of a four-step convergent synthesis illustrated in Scheme 1. Step 1 synthesizes the "methyl ester" intermediate AP25047 from starting materials AP24595, AP28141, and AP25570. Step 2 synthesizes the "aniline" intermediate AP24592 from starting material AP29089. Step 3 is the base-catalyzed coupling of AP25047 and AP24592 to produce ponatinib free base, also known as AP24534, which is isolated as the free base. Step 4 is the formation and crystallization of ponatinib monohydrochloride in ethanol.

[0321] A representative synthetic route to ponatinib HCl is designated Step C.

[0322] Scheme 1 :Process C [ka]

[0323] Step 1: Synthesis of AP25047 ("methyl ester") intermediate Overview and synthesis scheme Step 1 of the ponatinib HCl process is the synthesis of the methyl ester intermediate AP25047 from starting materials AP24595, AP25570, and AP28141 in three reaction sequences (designated 1a, 1b, and 1c) that eliminate intermediate isolation ("shorten"), as illustrated in Scheme 1. Two aromatic ring systems, juxtaposed and connected by a single alkyne linker, are constructed via two tandem palladium / copper-catalyzed Sonogashira couplings and an in situ desilylation reaction under basic conditions. The crude AP25047 product is then subjected to a series of processing steps designed to remove residual inorganic catalyst and treat by-products. These operations include crystallization of AP25047 as the HCl salt from the nonpolar solvent toluene (unit operation 1.3), aqueous workup and filtration through a silica gel plug (unit operation 1.4), and crystallization from the polar solvent 2-propanol (unit operation 1.5). These two crystallizations constitute an orthogonal purification to eliminate related-substance impurities of different polarity. The crystallization of the HCl salt from toluene and solvent washes are controlled by in-process analytical testing of specific process impurities. The final crystallization of the AP25047 intermediate from 2-propanol is being subjected to multivariate DoE testing to define the design space to ensure the exclusion of other impurities arising from the shortened reaction. A series of eight in-process tests in Phase 1 provide quantitative analytical control of reaction completion, exclusion of impurities, and effective removal of residual solvent.

[0324] Scheme 2 Step 1 - Synthesis of AP25047 [ka]

[0325] Unit Operation 1.1: First Sonogashira Reaction AP24595, palladium tetrakistriphenylphosphine (Pd(PPh3)4), copper(I) iodide (CuI), triethylamine, and tetrahydrofuran (THF) are placed in a reactor. The mixture is stirred and degassed with nitrogen, and then pre-degassed AP28141 is added. The resulting mixture is heated to 45-55°C and maintained for at least 3 hours. Completion of the reaction is determined by IPC-1 (HPLC). If the IPC-1 criteria are met, the mixture is concentrated to the desired volume and cooled.

[0326] Unit Operation 1.2: Deprotection / Second Sonogashira Reaction The reactor is charged with AP25570, additional palladium tetrakistriphenylphosphine (Pd(PPh3)4), copper(I) iodide (CuI), and tetrahydrofuran (THF). The mixture is concentrated and the water content is determined by IPC-2 (KF). If the IPC-2 criteria are met, the mixture is warmed to 45-60°C and a 25% solution of sodium methoxide in methanol is slowly added. The reaction mixture is stirred and held at 45-55°C for 30-60 minutes. The progress of the reaction is determined by IPC-3 (HPLC). The reaction mixture may be held at a lower temperature during IPC analysis. If the IPC-3 criteria are met, the process continues to unit operation 1.3.

[0327] Unit Operation 1.3: Isolation of AP25047 HCl The cooled reaction mixture is quenched by the addition of hydrogen chloride gas while stirring. Once a precipitate forms, residual hydrogen chloride is removed from the suspension by nitrogen purging. Tetrahydrofuran (THF) is replaced with toluene by azeotropic distillation under reduced pressure. The resulting warm slurry is filtered through an agitated filter dryer, and the cake is triturated and washed with toluene. The content of the process impurity AP29116 is determined by IPC-4 (HPLC). If IPC-4 criteria are met, the wet cake is dried by stirring under nitrogen flow and reduced pressure at 35-45°C (jacket temperature). Drying is monitored by IPC-5 (LOD, gravimetric). If IPC-5 criteria are met, crude AP25047 HCl is removed and packaged in FEP bags in plastic containers. The isolated AP25047 HCl can be stored for up to 7 days before further processing.

[0328] Unit operation 1.4: Post-processing The crude AP25047 HCl solid is placed in a container with dichloromethane (DCM) and washed with aqueous ammonia. The aqueous phase is back-extracted with DCM to recover yield, and the combined organic phases are washed a second time with aqueous ammonia. The organic layer is then washed with aqueous hydrochloric acid until the aqueous phase reaches a pH of 1-2 as indicated by IPC-6 (pH paper). If the IPC-6 criteria are met, the organic phase is treated with aqueous sodium bicarbonate until the aqueous wash reaches a pH of NLT7 as indicated by IPC-7 (pH paper). The organic phase is briefly concentrated, and fresh dichloromethane is added. The organic solution is passed through a silica gel pad and washed with fresh dichloromethane to increase product recovery.

[0329] Unit Operation 1.5: Crystallization of AP25047 The dichloromethane solution is concentrated under reduced pressure, and the dichloromethane is replaced with 2-propanol by azeotropic distillation under reduced pressure to the desired final volume range. The resulting suspension is then cooled and stirred for further aging.

[0330] Unit Operation 1.6: Isolation / Drying The precipitated product is isolated in an agitated filter dryer under nitrogen flow, and the cake is washed with 2-propanol. The wet cake is dried by agitation under nitrogen flow and vacuum at 45-55°C (jacket temperature). Drying is monitored by IPC-8 (LOD, gravimetric). If IPC-8 criteria are met, the product is collected and packaged in polyethylene bags, which are then placed in heat-sealed Mylar-coated aluminum foil bags and placed in HDPE shipping containers (expected yield range: 65-89%).

[0331] Step 2: Synthesis of AP24592 ("aniline") intermediate Overview and synthesis scheme Step 2 of the ponatinib HCl process, illustrated below, is the synthesis of the aniline intermediate AP24592 by catalytic hydrogenation of the nitro-aromatic starting material AP29089. The reaction is carried out in ethyl acetate, a solvent in which the starting materials and product are highly soluble. The catalyst for this reaction is palladium on carbon, and hydrogen is introduced as a gas directly into the reaction mixture. Upon completion of the reaction, the solvent is switched from ethyl acetate to n-heptane by distillation, which promotes spontaneous crystallization of AP24592, yielding a highly purified material. This crystallization has been shown to be highly effective in purifying the product, as most of the process impurities remain solubilized in n-heptane.

[0332] The three in-process controls for Stage 2 are HPLC of the reaction mixture to confirm consumption of starting material, GC measurement of ethyl acetate after azeotropic solvent exchange to n-heptane, and gravimetric measurement of solvent loss upon drying.

[0333] Scheme 3 Step 1 to Step 2: Synthesis of AP24592 [ka]

[0334] Unit Operation 2.1: Dissolution and Hydrogen Purging AP29089, 10% palladium on carbon and ethyl acetate are placed in a reactor and the suspension is stirred under hydrogen pressure.

[0335] Unit Operation 2.2: Hydrogenation After pressurizing the reactor with hydrogen until a stable pressure range is reached, the mixture is stirred under a hydrogen atmosphere for at least 4 more hours. The reactor is depressurized and samples are taken to assess the completion of the reaction (IPC-1). If the IPC-1 criteria are met, the process continues to unit operation 2.3.

[0336] Unit Operation 2.3: Concentration / Crystallization The reaction mixture is passed through a filter cartridge to remove the catalyst, and the cartridge is washed with fresh ethyl acetate. The combined filtrate and wash solution are concentrated under vacuum to remove the desired volume of ethyl acetate. n-Heptane is added and distillation is continued under vacuum to the target volume. The ethyl acetate content is measured by IPC-2 (GC). If the IPC-2 criteria are met, the process continues to unit operation 2.4.

[0337] Unit Operation 2.4: Isolation / Drying The solid product is dried under vacuum at the target temperature range. Completion of drying is determined by IPC-3 (LOD, gravimetric). AP24592 is obtained in the range of 80-97% (based on the AP29089 input) as a white to yellow solid.

[0338] Step 3: Synthesis of ponatinib free base Overview and synthesis scheme Step 3, shown in Scheme 4, is the synthesis of ponatinib free base via the base-catalyzed reaction of AP25047 with AP24592. This reaction is carried out in the presence of the strong base potassium tert-butoxide under substantially anhydrous conditions to minimize undesired hydrolysis of the methyl ester of AP25047 to the corresponding unreactive carboxylic acid. The presence of this by-product not only reduces yield but also complicates downstream processing during reaction workup. Drying of the reaction mixture via a series of azeotropic distillations ensures reliable reaction and quantitative consumption of starting materials by monitoring and controlling water in the process. Reaction conditions and crystallization parameters that ensure the elimination of process impurities are well understood based on DoE studies.

[0339] Scheme 4 Step 3 - Synthesis of AP24534 Free Base [ka]

[0340] Unit Operation 3.1: Drying of the Reaction Mixture AP25047, AP24592, and 2-methyltetrahydrofuran (2-Me-THF) are placed in a reactor. The mixture is concentrated under reduced pressure to the target volume. Fresh 2-methyltetrahydrofuran is added and distillation is performed again. After another 2-methyltetrahydrofuran addition and distillation cycle, the water content of the mixture is measured by IPC-1 (KF). If the IPC-1 criteria are met, the process continues to unit operation 3.2.

[0341] Unit Operation 3.2: Reaction Potassium tert-butoxide (KOtBu) is added while stirring the suspension and maintaining the target temperature between 13 and 23°C. After 3 hours or more, the reaction progress is assessed by HPLC (IPC-2). If the IPC criteria are met, the process continues to unit operation 3.3.

[0342] Unit Operation 3.3: Quenching and Extraction The reaction mixture is diluted with 2-methyltetrahydrofuran (2-Me-THF) and quenched by the addition of aqueous sodium chloride. The organic layer is separated, and the aqueous layer is extracted twice with 2-methyltetrahydrofuran. The combined organic layer is washed sequentially with aqueous sodium chloride and water. The organic layer is then aged at 15-30°C.

[0343] Unit Operation 3.4: Concentration / Solvent Exchange After aging (see unit operation 3.3), the mixture is passed through a cartridge filter and concentrated under vacuum to the target volume. 1-Propanol is added and stirred at elevated temperature to form a solution, which is then distilled under vacuum to the target volume and gradually cooled to a temperature range of 20-30°C.

[0344] Unit Operation 3.5: Crystallization The product solution in 1-propanol is aged with stirring at a temperature of 20-30°C until the presence of solids is visually observed. Acetonitrile is added to the suspension with stirring, and the resulting suspension is further aged at 20-30°C for 60-120 minutes before isolation in the next unit operation.

[0345] Unit Operation 3.6: Isolation / Drying The slurry produced in unit operation 3.5 is isolated under vacuum in a filter / dryer. The solids are washed twice with a mixture of 1-propanol and acetonitrile. The solids are then dried under vacuum and monitored by IPC-3 (LOD, gravimetric). If IPC criteria are met, the product is discharged as an off-white to yellow solid, bagged in double polyethylene bags, and stored at ambient temperature.

[0346] Step 4: Synthesis of Ponatinib HCl Overview and synthesis scheme Step 4 of the ponatinib HCl process involves combining equimolar amounts of ponatinib free base with hydrochloric acid in ethanol to form the monohydrochloride salt and inducing crystallization by seeding. The parameters of this process have been investigated in DoE studies for their effect on the production of the desired solid form and the particle size distribution of this process. The synthesis scheme for Step 4 is shown in Scheme 5.

[0347] Scheme 5 Step 4 - Synthesis of Ponatinib HCl [ka]

[0348] Unit Operation 4.1: Dissolution Place AP24534 free base and absolute ethanol (EtOH) in a reactor and stir at 60-75°C to form a solution. Confirm dissolution by visual observation.

[0349] Unit Operation 4.2: Clarification The solution is passed through a filter and then washed with ethanol at 60-78°C.

[0350] Unit Operation 4.3: Acidification / Seeding The product solution is concentrated under vacuum to the target volume. With stirring, the first portion (approximately 25%) of a 1N ethanolic solution of hydrogen chloride is added to the reactor. The solution is seeded with qualified AP24534HCl crystals at a temperature of 60-70°C to initiate crystallization. The process continues through unit operation 4.4.

[0351] Unit Operation 4.4: Crystallization If visual observation reveals the presence of solids in the reactor, slowly add the remaining 1N ethanolic hydrogen chloride solution (approximately 75%) to the stirred mixture. Allow the mixture to age for at least 10 minutes and perform IPC-1 to measure the pH of the solution. If the IPC criteria are met, cool the mixture to a temperature of 5-15°C and age it with stirring.

[0352] Unit Operation 4.5: Isolation / Drying The solid product is isolated by filtration and washed with ethanol at a temperature of 5-15°C. Excess ethanol is removed from the solid with gentle agitation and a nitrogen stream at ambient temperature. The solid is then dried under vacuum at 60-70°C. Drying is monitored by IPC-2 (LOD, gravimetric). If IPC-2 criteria are met, ponatinib HCl is removed as an off-white to yellow solid, packaged in double polyethylene bags, and stored in plastic drums at 20-30°C.

[0353] It should be understood that the foregoing description is exemplary and explanatory in nature, and is intended to describe the general inventive concepts of the present disclosure and its preferred embodiments. Those skilled in the art having the benefit of this disclosure may recognize, through routine experimentation, obvious modifications and variations without departing from the spirit and scope of the present disclosure. Accordingly, the present disclosure is not limited to the above description, but rather by the following claims and their equivalents.

Claims

1. A polymorphic crystal of crystalline Form F of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride, characterized by a powder X-ray diffraction pattern containing a peak at at least one 2θ value (±0.3°) selected from the following angles (°): 6.8; 9.8; 12.4; 16.2; 17.9; 19.0; 24.0; and 25.

1.

2. 2. The polymorphic crystal of crystalline Form F of claim 1, wherein said polymorphic crystal of crystalline Form F is substantially pure.

3. A pharmaceutical composition comprising the crystalline Form F polymorph of any one of claims 1 and 2 and a pharmaceutically acceptable carrier.

4. A pharmaceutical composition comprising the crystalline Form F polymorph of any one of claims 1 and 2 and a pharmaceutically acceptable carrier or excipient.

5. A therapeutic agent for treating chronic myeloid leukemia (CML) or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL), comprising a therapeutically effective amount of the crystalline form F polymorph of any one of claims 1 and 2 or the pharmaceutical composition of claim 3 or 4.

6. Use of the polymorphic crystals of crystalline Form F according to any one of claims 1 to 2 in the manufacture of a medicament for treating chronic myeloid leukemia (CML) or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL).

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