AU2016210725A1 - Crystalline forms of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride - Google Patents

Abstract

Novel crystalline forms of 3-(imidazo[1,2-blpyridazin-3-ylethyny)-4-methyl-N-{4-[(4 methylpiperazin- 1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride and to methods of their preparation are disclosed herein.

Description

CRYSTALLINE FORMS OF

3-(IMlDAZO[1,2-B]PYRIDAZIN-3-YLETHYNYL)-4-METHYL-N‘{4-[{4"METHYLPIPERAZIN"1-YL)METHYL]-3-(TRIFLUOROMETHYL)PHENYL>BENZAMIDE MONO HYDROCHLORIDE

This is a divisional of Australian patent application No. 2013204506, the entire contents of which are incorporated herein by reference.

BACKGROUND

[0001] The instant application is directed 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 mono hydrochloride and to methods of their preparation.

[0002] 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride has the chemical formula Cz9H2eCIF3NeO which corresponds to a formula weight of 569.02 g/mol. Its chemical structure is shown below:

The CAS Registry number for 3-(!midazo[1,2-b]pyridazin-3-y!ethynyl)-4-methy[-N-{4-[{4-methylpiperazin~1-yl)methyl]-3-(!rifluoromethyl)phenyI}benzamide mono hydrochloride is 1114544-31-8.

[0003] The United States Adopted Name (USAN) and international Nonproprietary Name (INN) of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yi)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is ponatinib hydrochloride. Alternative chemical names for ponatinib hydrochloride include benzamide, 3-{2-imidazo[1.2-b]pyridazin-3-ylethynyl)-4-methyl-/V-[4-[(4-methyl“1“ piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-, hydrochloride (1:1) and 3-[2-(imidazo[1,2-jb]pyridazin-3-yl)ethynyl]-4-methyl-/V-{4-[(4-methylpiperazin-1- yl)methyl]-3-(trifluoromethyl)phenyl}benzamide monohydrochloride.

[0004] Ponatinib hydrochloride is a small molecuie pan-BCR-ABL inhibitor in clinical development for the treatment of adult patients with chronic phase, accelerated phase, or blast phase chronic myeloid leukemia (CML) or Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ ALL) resistant or intolerant to prior tyrosine kinase inhibitor therapy. Other tyrosine kinase inhibitors relevant to such CML or Ph+ALL therapy include GLEEVEC® (imatinib mesylate) and TASIGNA® (nilotinib) (both from Novartis AG), SPRYCEL® (dasatinib) (from Bristol Myers Squibb Company) and BOSULIF® (bosutinib) (from Pfizer Inc). A New Drug Application for ponatinib hydrochloride was filed with the United States FDA on July 30, 2012, [0005] In addition, ponatinib hydrochloride is potentially clinically useful for the treatment of other disorders or conditions implicated by the inhibition of other protein kinases. Such kinases and their associated disorders or conditions are mentioned in O'Hare, T., ef at., Cancer Cell, Volume 16, Issue 5, 401-412 (2009) and WO 2011/053938, both of which are hereby incorporated herein by reference for all purposes.

[0006] Having an understanding of the potential polymorphic forms for an active pharmaceutical ingredient (API) such as ponatinib hydrochloride is useful in the development of a drug. This is because not knowing the specific polymorphic form present or desired in the API may result in inconsistent manufacturing of the API and as a result, results with the drug may vary between various lots of the API. In addition, it is important to discover the potential polymorphic forms of an API so that one can systematically determine the stability of that form over a prolonged period of time for similar reasons. Once a specific polymorphic form is selected for pharmaceutical development, it is important to be able to reproducibly prepare that polymorphic form. It is also desirable for there to be a process for making an API such as ponatinib hydrochloride in high purity due to the potential of impurities to affect the performance of the drug.

[0007] The earliest patent publication known by Applicant to disclose the chemical structure of ponatinib hydrochloride is WO 2007/075869, which is also owned by Applicant (ARIAD Pharmaceuticals, Inc.) and is hereby incorporated herein by reference for all purposes. Example 16 of WO 2007/075869 states that the product was obtained as a solid: 533 m/z (M+H), This mass corresponds to the free base of ponatinib. Example 16 also discusses the preparation of a mono hydrochloride salt of ponatinib. Example 16 neither specifically mentions that the ponatinib hydrochloride obtained was crystalline nor specifies any particular crystalline forms of ponatinib hydrochloride.

[0008] United States Serial No. 11/644,849, which published as US 2007/0191376, is a counterpart application to WO 2007/075869 and granted on February 14, 2012 as U.S. Patent No. 8,114,874, which is hereby incorporated herein by reference for all purposes. United States Serial No. 13/357,745 is a continuing application of USSN 11/644,849, which also is hereby incorporated herein by reference for all purposes.

[0009] Additional patent applications owned by Applicant that cover ponatinib hydrochloride and published as of the filing date of this application include WO 2011/053938 and WO 2012/139027, both of which are hereby incorporated herein by reference for all purposes. Like WO 2007/075869, neither of WO 2011/053938 or WO 2012/139027 specifies any particular crystalline forms of ponatinib hydrochloride.

SUMMARY

[0010] It has now been discovered that ponatinib hydrochloride can exist in certain crystalline forms, certain of which are suitable for tablet development.

[0011] In one aspect, the present disclosure is directed to substantially pure crystalline forms of ponatinib hydrochloride. The substantially pure crystalline form of ponatinib hydrochloride is Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form I or Form J.

[0012] In another aspect, the present disclosure is directed to pharmaceutical compositions comprising a therapeutically effective amount of a substantially pure crystalline form of ponatinib hydrochloride disclosed herein and at least one pharmaceutically acceptable carrier, vehicle or excipient.

[0013] In another aspect, the present disclosure is directed to a method of treating a disorder or condition in a human 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, the disorder or condition is chronic myeloid leukemia (CML) or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ALL) when the protein kinase is Bcr-Abl or a mutant form thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present inventions. The inventions may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0015] FIG. 1 is a summary of the eleven solid forms of ponatinib hydrochloride that include HCI polymorphs and pseudo-polymorphs that are identified as Forms A through K and that were discovered and are disclosed herein.

[0016] FIG. 2 is a summary of certain of the solid forms of ponatinib hydrochloride identified in Figure 1 that were discovered and are disclosed herein. The legend for Figure 2 is as follows: a Starting material: Form HCI1 or amorphous material (Am) obtained by freeze-drying. b Occ: the total occurrence included 216 experiments carried out in Phase 2 for which 39 samples were analyzed additionally wet or the mother liquor was evaporated and analyzed giving a total of 254 materials characterized. For example, “(3, 1.2%)" correspond to 3 occurrences of the form out of 254 measurements, giving a percentage of 1.2%. For 62 out of the 254 measurements (9%), the product yield was too low to identify the solid form, or the materials were wet. d Am: amorphous form.

[0017] FIG. 3 is a characteristic X-ray powder diffraction (XRPD) pattern of two batches of Form A of ponatinib hydrochloride in which the data for each batch was acquired prior to and after DVS humidity cycling. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0018] FIG. 4 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form A of ponatinib hydrochloride. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0019] FIG. 5 is a characteristic differential scanning calorimetry (DSC) scan obtained from Form A of ponatinib hydrochloride. Pleat flow [mW] is shown on the vertical axis and temperaiure (°C) is shown on the horizontal axis, [0020] FIG, 6 is a characteristic thermogravimetric analysis (TGA) and thermogravimetric analysis with mass spectroscopic analysis of volatiles (TGMS) scan obtained from Form A of ponatinib hydrochloride.

[0021] FIG. 7 is a characteristic 1H-NMR Spectrum (600 MHz) obtained from Form A of ponatinib hydrochloride in DMSO-d6 at 300 K. Normalized Intensity is shown on the vertical axis and chemical shift (ppm) is shown on the horizontal axis, [0022] FIG. 8 is a characteristic 19F-NMR Spectrum (564 MHz) obtained from Form A of ponatinib hydrochloride in DIVISO-d6 at 300 K. Normalized intensity is shown on the vertical axis and chemical shift (ppm) is shown on the horizontal axis, [0023] FIG, 9 is a characteristic 13C-NMR Spectrum (151 MHz) obtained from Form A of ponatinib hydrochloride in DMSO-de at 300 K. Normalized Intensity is shown on the vertical axis and chemical shift (ppm) is shown on the horizontal axis.

[0024] FIG, 10 is a characteristic mass spectral pattern obtained from Form A of ponatinib hydrochloride in which the top mass spectral pattern is the observed mass of Form A and the bottom mass spectral pattern is the calculated mass of Form A. Relative abundance is shown on the vertical axis and atomic weight (m/z) is shown on the horizontal axis.

[0025] FIG. 11 is a characteristic mass spectral fragmentation pattern of Form A of ponatinib hydrochloride. Relative abundance is shown on the vertical axis and atomic weight (m/z) is shown on the horizontal axis.

[0026] FIG. 12 shows the structure of Form A of ponatinib hydrochloride in accordance with the data presented in the table herein designated as “Crystal Data and Structure Refinement for Ponatinib Hydrochloride Form A.” Atoms in this Figure 12 are color coded according to atom type; carbon, grey; nitrogen, blue; oxygen, red; hydrogen, white; fluorine, yellow; chlorine, green, [0027] FIG, 13 is a characteristic FT-IR spectrum obtained from Form A of ponatinib hydrochloride. Percent transmittance (%) is shown on the vertical axis and wavenumber (cm'1) is shown on the horizontal axis.

[0028] FIG. 14 is a characteristic HPLC spectrum obtained from Form A of ponatinib hydrochloride, Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis, [0029] FIG, 15 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form B of ponatinib hydrochloride as compared against a XRPD pattern of Form A and Form C. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0030] FIG. 16 is a characteristic HPLC spectrum obtained from Form B of ponatinib hydrochloride. Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

[0031] FIG. 17 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form C of ponatinib hydrochloride as compared against a XRPD pattern of Form A. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0032] FIG. 18 is a characteristic differential scanning calorimetry (DSC) scan obtained from Form C of ponatinib hydrochloride. Heat flow [mWJ is shown on the vertical axis and temperature (°C) is shown on the horizontal axis.

[0033] FIG. 19 is a characteristic thermogravimetric analysis (TGA) scan obtained from Form C of ponatinib hydrochloride.

[0034] FIG. 20 is a characteristic TGMS thermogram obtained from Form C of ponatinib hydrochloride, [0035] FIG. 21 is a characteristic HPLC spectrum obtained from Form C of ponatinib hydrochloride. Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

[0036] FIG. 22 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form D of ponatinib hydrochloride as compared against a XRPD pattern of Form A and certain other crystalline forms within the class of HCI3. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0037] FIG. 23 is a characteristic differential scanning calorimetry (DSC) scan obtained from Form D of ponatinib hydrochloride. Heat flow [mW] is shown on the vertical axis and temperature (°C) is shown on the horizontal axis, [0038] FIG. 24 is a characteristic thermogravimetric analysis (TGA) scan obtained from Form D of ponatinib hydrochloride, [0039] FIG. 25 is a characteristic FT-IR spectrum obtained from Form D of ponatinib hydrochloride. Percent transmittance (%) is shown on the vertical axis and wavenumber (cm'1) is shown on the horizontal axis. The Form A starting material is shown in red and Form D (PSM1) is shown in blue.

[0040] FIG. 26 is a characteristic HPLC spectrum obtained from Form D of ponatinib hydrochloride. Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

[0041] FIG. 27 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form F of ponatinib hydrochloride as compared against a XRPD pattern of Form A and certain other crystalline forms within the class of HCI5. Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis, [0042] FIG. 28 shows two characteristic differential scanning calorimetry (DSC) scans obtained from Form F of ponatinib hydrochloride. The top scan is the DSC curve of VDS1. The bottom scan is the DSC curve of VDS2. Heat flow [mW] is shown on the vertical axis and temperature (°C) is shown on the horizontal axis, [0043] FIG. 29 is a characteristic thermogravimetric analysis (TGA, top) and TGMS (bottom) scan obtained from Form F of ponatinib hydrochloride (VDS1).

[0044] FIG. 30 is a characteristic thermogravimetric analysis (TGA, top) and TGMS (bottom) scan obtained from Form F of ponatinib hydrochloride (VDS2).

[0045] FIG. 31 is a characteristic FT-IR spectrum obtained from Form F of ponatinib hydrochloride. Percent transmittance (%) is shown on the vertical axis and wavenumber (cm'1) is shown on the horizontal axis. The Form A starting material is shown in red and Form F (VDS1) is shown in green.

[0046] FIG. 32 is a characteristic FT-IR spectrum obtained from Form F of ponatinib hydrochloride. Percent transmittance (%) is shown on the vertical axis and wavenumber (cm '1) is shown on the horizontal axis. The Form A starting material is shown in purple and Form F (VDS2) is shown in red.

[0047] FIG. 33 is a characteristic HPLC spectrum obtained from Form F of ponatinib hydrochloride (VDS2). Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

[0048] FIG. 34 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form H of ponatinib hydrochloride as compared against a XRPD pattern of Form A and certain other crystalline forms within the class of HCI6. Relative Intensity {in counts} is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0049] FIG. 35 shows two characteristic differential scanning calorimetry (DSC) scans obtained from Form FI of ponatinib hydrochloride. The top scan is the DSC curve of VDS3. The bottom scan is the DSC curve of VDS4, Heat flow [mW] is shown on the vertical axis and temperature (°C) is shown on the horizontal axis, [0050] FIG. 36 is a characteristic thermogravimetric analysis (TGA, top) and TGMS (bottom) scan obtained from Form FH of ponatinib hydrochloride (VDS3), [0051] FIG. 37 is a characteristic thermogravimetric analysis (TGA, top) and TGMS (bottom) scan obtained from Form FI of ponatinib hydrochloride (VDS4).

[0052] FIG. 38 is a characteristic FT-iR spectrum obtained from Form H of ponatinib hydrochloride, Percent transmittance (%) is shown on the vertical axis and wavenumber (cnrf1) is shown on the horizontal axis. The Form A starting material is shown in purple and Form H (VDS3) is shown in red.

[0053] FIG, 39 is a characteristic FT-IR spectrum obtained from Form FI of ponatinib hydrochloride. Percent transmittance (%) is shown on the vertical axis and wavenumber (cm-1) is shown on the horizontal axis. The Form A starting material is shown in purple and Form FI {VDS4) is shown in red.

[0054] FIG. 40 is a characteristic FIPLC spectrum obtained from Form FI of ponatinib hydrochloride (VDS4). Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

[0055] FIG. 41 is an overlay of characteristic X-Ray Powder Diffraction (XRPD) patterns for each of the solid forms identified in Figure 1 where the vertical axis denotes relative intensity (counts) and the horizontal axis denotes Two Theta (Degrees). From the bottom to the top of this figure the following solid forms and solvents are as follows: the starting material ponatinib HCI (HCI1), Form FICI2 (QSA12.1, solvent: water), Form HCI2b(QSA21.1, solvent: water), Form FHC|3-class (GRP12.1, solvent: toluene), mixture HCI1+FIC14 (GRP1.1, solvent: hexafluorobenzene), Form FICI5 (VDS28.1, solvent: butylacetate), Form HCISb (VDS28.2 after drying, solvent: butylaceiaie} and HCI6-class (VDS6.1, solvent: methanol).

[0056] FIG. 42 shows representative digital images of (from top to bottom, left to right): HCI6-class (VDS6.1, vds050.0c:E1), Form HCI5 (VDS28.1, vds05.0c:B3). Form HCI5b (VDS28.2, vds05.1c:B6), mixture HCI1+HC14 (GRP1.1, grp02.0c:A1), Form HCI3-class (GRP12.1, grp02.1c:L1), Form HCI2 (QSA12.1, qsa00.1c:A2) and Form HCl2b (QSA21.1. qsa00.1c:J2).

[0057] FIG. 43 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form J of ponatinib hydrochloride. Relative Intensity (ip counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0058] FIG. 44 is a characteristic X-Ray Powder Diffraction (XRPD) pattern obtained from Form K of ponatinib hydrochloride, Relative Intensity (in counts) is shown on the vertical axis and the degrees (2Θ) is shown on the horizontal axis.

[0059] FIG. 45 shows XRPD patterns of Form A of ponatinib hydrochloride (botlom pattern) and amorphous ponatinib hydrochloride (top pattern) (solvents,2,2-trifluroethano|) where the vertical axis denotes relative intensity (counts) and the horizontal axis denotes Two Theta (Degrees).

[0060] FIG. 46 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 mono hydrochloride. An intense endothermic . event with a peak of 259.4°C was observed, corresponding to the melting point of the amorphous form. The vertical axis denotes Heat Flow [mW] and the horizontal axis denotes Temperature (°C).

[0061] FIG. 47 is a characteristic HPLC spectrum obtained from Form H of ponatinib hydrochloride (VDS4). Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis. Absorbance units are shown on the vertical axis (mAU) and time (minutes) is shown on the horizontal axis.

DETAILED DESCRIPTION OF THE INVENTION

[0062] It was surprisingly discovered that 3-(imidazo[1,2~b]pyridazin-3-ylethynyl)~4- methyl-N-{4“[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)pheny|}benzamlde mono hydrochloride can be obtained in various solid state crystalline forms. "Crystalline form" or “polymorphic form" or “polymorph", as these terms may be used interchangeably herein, refers to a crystalline form of ponatinib hydrochloride that is distinct from the amorphous form of ponatinib hydrochloride and other crystalline form(s) of ponatinib hydrochloride as determined by certain physical properties such thermodynamic stability, physical parameters, x-ray structure, DSC and preparation processes. While polymorphism classically refers to the ability of a compound to crystallize into more than one distinct crystal species (having Identical chemicai structure but quite different physicochemical properties), the term pseudopolymorphism is typically applied to solvate and hydrate crystalline forms. For purposes of this disclosure, however, both true polymorphs as well as pseudopolymorphs, i.e., hydrate and solvate forms, are included in the scope of the term “crystalline forms" and “polymorphic forms." In addition, "amorphous" refers to a disordered solid state. It should be noted that different samples of a particular crystalline form will share the same major XRPD peaks, but that there can be variation in powder patterns with regard to minor peaks. In addition, the term "about" with regard to XRPD maxima values {in degrees two theta) generally means within 0.3 degrees two theta, of the given value; alternatively, the term "about" means (in this and all contexts) within an accepted standard of error of the mean, when considered by one of ordinary skill in the art. As used herein, the terms "isolated" and "substantially pure" mean more than 50% of the crystalline ponatinib hydrochloride is present {as can be determined by a method in accordance with the art) in the identified crystalline form relative to the sum of other solid form(s) present in the selected material. DEFINITIONS AND ABBREVIATIONS Solvent abbreviation: • DMA N,N-Dimethylacetamide • DMF N.N-Dimethylformamide • DMSO Dimethylsulfoxide . TFE 2,2,2-Trifluoroethanol • THF Tetrahydrofuran • E10H Ethanol • MeOH Methanol

Other abbreviations (alphabetical order): • Am Amorphous • API Active Pharmaceutical Ingredient • AS Anti-solvent • Cl Counter-ion • DSC Differential Scanning Calorimetry • DVS Dynamic Vapor Sorption • GRP ID for Grinding experiment • HPLC High-Performance Liquid Chromatography • MS Mass Spectroscopy • PSM ID for Cooling/evaporative crystallization experiment • SAS Solubility assessment • SDTA Single Differential Thermal Analysis • S Solvent • SM Starting material • TGA Thermogravimetric Analysis • TGMS Thermogravimetric Analysis coupled with Mass Spectroscopy • VDL ID for Vapor diffusion into liquids experiments • VDS ID for Vapor diffusion onto solids experiments » XRPD X-Ray Powder Diffraction [0063] Through XRPD analysis, a total of eleven polymorphic forms of ponatinib hydrochloride were discovered. Each of the eleven new polymorphic forms are referred to herein as: HCI1 (also referred to herein as “Form A"), HCI2 (also referred to herein as "Form B"), HCI2b (also referred to herein as "Form C"), HCI3-class (also referred to herein as "Form D"), a mixture HCI1+HCS4 (also referred to herein as "Form E"), HCI5-class or simply HCI5 (also referred to herein as "Form F"), HCISb or HCI desolvate (also referred to herein as "Form G"), HCl6-class (also referred to herein as "Form H"), HCI6 desolvate (also referred to herein as "Form I”), HCI7 (also referred to herein as "Form J"), and HCI8 (also referred to herein as "Form K"). The nature or origin of these eleven polymorphic forms is indicated in Figure 1. In addition, certain characteristics of the referenced polymorphic form are provided as well. For instance,

Form A is indicated as being an anhydrate of ponatinib hydrochloride and additionally was obtained as a single crystal.

[0064] In general, crystalline forms of ponatinib hydrochloride have physical properties (such as high stability, etc.) that are advantageous for the commercial preparation of solid dosage forms as compared to amorphous ponatinib hydrochloride. The distinction between crystalline ponatinib hydrochloride and amorphous ponatinib hydrochloride can be readily seen with the same type of physical chemical data (e.g., DSC, XRPD, thermal analysis) that is used to distinguish the individual crystalline forms of ponatinib hydrochloride disclosed herein.

[0065] With reference to the foregoing methodologies, attention is now drawn to each of the discovered polymorphs of 3-(irnidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride.

[0066] Characteristics of Form A fHCH): [0067] The anhydrate HCI1 (same crystalline form as the starting material) was the predominant crystalline form discovered. The chemical structure of ponatinib hydrochloride has been unambiguously established by a combination of nuclear magnetic resonance spectroscopy (NMR), mass spectrometry (MS), and single crystal X-ray crystallography with confirmatory data from elemental and chloride analysis, Fourier transform infra-red (FT-IR) spectroscopy, and ultraviolet (UV) spectroscopy. The preferred solid form of ponatinib hydrochloride is the anhydrous crystalline HCI-1 solid form or Form A.

[0068] With reference to Figure 3, samples of Ponatinib HCI, ASI Batch 110020 and CGAM Batch F08-Q6Q57 were analyzed by X-ray powder diffraction (XRPD). In each case, the materiai was analyzed prior to and after DVS humidity cycling. XRPD patterns were obtained using a high-throughput XRPD diffractometer. Data collection was carried out at room temperature using monochromatic CuKa radiation in the 2Θ region between 1.5° and 41.5°, which is the most distinctive part of the XRPD pattern. The diffraction pattern of each well was collected In two 2Θ ranges (1.5° 5 2Θ £ 21.5° for the first frame, and 19.5° s 2Θ i 41.5° for the second) with an exposure time of 90 seconds for each frame. No background subtraction or curve smoothing was applied to the XRPD patterns. Figure 3 shows the X-ray powder diffraction pattern of these materials, each in the HCI-1 solid form, This powder pattern is consistent with the powder pattern simulated from single crystal X-ray diffraction experiments on the HCI-1 form. XRPD data acquired prior to, and after DVS humidity cycling experiments, indicates that the HCI-1 solid form is maintained after humidity cycling. In the XRPD pattern of Form A shown in Figure 3, at least one or all of the following peaks in degress two theta (2Θ) is shown: 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 shows two peaks, three peaks, four peaks or five peaks. The term "about" applies to each listed peak for this and all other forms mentioned in this disclosure.

[0069] Figure 4 shows a characteristic X-Ray Powder Diffraction (XRPD) pattern for Form A of ponatinib hydrochloride in which greater detail relative to the XRPD is seen. The XRPD pattern of Form A shown in Figure 4 shows at least one or more of the following peaks in degress 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, the XRPD pattern of Form A shows two peaks, three peaks, four peaks or five peaks.

[0070] In differential vapor sorption (DVS) experiment with HCI-1, the relative humidity was cycled from 45% to 95% (sorption), to 0% (desorption) and back to 45% (sorption) at a constant temperature of 25°C, with a hold time of 60 minutes per step. The results of this DVS experiment on ponatinib HCI CGAM Batch F08-060507 exhibited a 1.1% water uptake at 95% humidity, and ponatinib HC! ASI Batch 110020 exhibited a 1.4% water uptake at a relative humidity of 95%. This water uptake was reversible on cycling to lower humidity. These results demonstrate that HCI-1 is not a hygroscopic compound, The effect of the humidity cycling on HCI-1 was also assessed by X-ray powder diffraction (XRPD) analysis before and after the DVS experiment. The XRPD data revealed that humidity cycling had no effect on the solid form of the material, which remained in the HCI-1 solid form.

[0071] With reference to Figure 5, the melting point of ponatinib HCI in the HCI-1 solid form, was determined by differential scanning calorimetry (DSC). The sample of ponatinib HCI, ASI Batch 110020, was analyzed in a pin-holed crucible in the temperature range of 25°C to 30Q°C at a heating rate of 1Q°C per minute using dry gas purge, An intense endothermic event with a peak of 264.1°C was observed, corresponding to the melting point of Form A.

[0072] With reference to Figure 6, Thermogravi metric analysis (TGA) and thermogravimetric analysis with mass spectroscopic analysis of volatiles (TGMS) was performed on ponatinib HCI, ASI Batch 110020. The sample, contained in a pin-holed crucible, was heated in the TGA instrument from 25°C to 300°C at a heating rate of 10°C min-1, with dry Nz gas used for purging. Gases evolved from the TGA were analyzed using a quadrupole mass spectrometer. Ponatinib HCI, ASI Batch 110020, in the HCI-1 solid form, contained 0.31% water by weight and 0.85% ethanol by weight at the time of release.The TGA/TGMS experiment indicated that mass tosses of 0.2% (water) and 0.6% (ethanol, from the crystallization solvent) are observed between the temperature range of 25-130°C and 130 - 240°C, respectively. These losses are consistent with the water and ethanol content at the time of release. Ethanol is released from the material at a higher temperature than water, although not associated with ponatinib HCI in the HCI-1 solid form as a solvate.

[0073] Extensive solution phase NMR studies using a combination of multiple 1D and 2D NMR methods were performed on Form A of ponatinib HCI to obtain a complete assignment of 1H, 1SF, and 13C resonances, and hence to confirm the chemical structure of ponatinib HCI. Analyses were performed at ARIAD Pharmaceuticals, Inc., Cambridge, MA, on a sample of ponatinib HCI (ASI Batch 110020) dissolved in deuterated DMSO (DMSO-dB) solvent. NMR spectra were acquired at a temperature of 300 K on a BruKer Avance III-600 MHz NMR spectrometer equipped with a 5 mm BBFO z-gradient probe. All 1H chemical shifts were referenced to the DMSO peak at 2.5 ppm. With reference to Figure 7, the 1D 1H-NMR spectra of Form A of ponatinib HCI in DMSO-d6 is shown. 1H resonance 32a arises from the protonated piperazine moiety in ponatinib HCI. The EtOH resonances appearing in both Ή (Figure 7} and 13C spectra (Figure 9) arise from residual EtOH present in ponatinib HCI, Figure 8 shows the 1D 19F-NMR spectra of Form A of ponatinib HCI in DMSO-d6 with a characteristic chemical shift at 57.94 ppm. Figure 9 shows the 1D 13C-NMR spectra of Form A of ponatinib HCI in DMSO-dB.

[0074] Table 1 summarizes the relevant chemical shift data of Form A obtained from the 1H and 13C-NMR experiments. The number of signals and their relative intensity (integrals) confirm the number of protons and carbons in the structure of Form A of ponatinib HCI. These chemical shift data are reported in according to the atom numbering scheme shown immediately below:

Table 1: 1H and 13C Chemical Shift Data (in ppm) of Form A of ponatinib HCI, in DMSO-ds at 300 K

m:multiplet dd: doublet οΓdoublets didoublet s: singlet br.s:broad singlet q: quartet ND: not determined due to spectral overlap in the 'H NMR spectrum.

Due to symmetry, the resonance pair 30 and 3d as well as the resonance pair 31 and 33 have degenerate chemical shills. In addition the methylene protons of these resonances appear as diaslereolopic pairs.

[0075] With reference to Figure 10, mass spectral experiments and collisionally activated MS2 fragmentation of Form A of ponatinib FiCI were carried out using Thermo Finnegan Exactive accurate mass and LTQ XL ion trap mass spectrometers, each operating in positive ion electrospray mode. Samples of Form A of ponatinib HCI (ASI Batch 110020), dissolved in acetonitrile, were introduced into the mass spectrometers via infusion by a syringe pump. The accurate mass for ponatinib HCI was obtained on the Exactive mass spectrometer using full scan mode. The mass observed in this infusion experiment is m/z 533.2269 (MH+) with the calculated exact mass being 533.2271 (MH+) yielding a mass difference of 0.2 mmu (Δ ppm of -0.38) (Figure 10 top). The fragmentation spectrum of ponatinib HCI on the Exactive mass spectrometer is shown in Figure 10, and contains the product ions from m/z of 533.2269 (the molecular ion of ponatinib HCI), as well as ions from any other co-eluted compounds.

[0076] Figure 11 shows MS fragmentation data obtained on the LTQ XL ion trap mass spectrometer. Figure 11(A) shows the full scan MS of m/z 533, (MH+) of the sample during infusion. Figure 11(B) (MS2 scan) shows the fragment spectrum of the selected mass m/z 533. Figure 11(C) and Figure 11(D) show the product ions from m/z 433 and 260 respectively; ions m/z 433, and 260 were themselves the Initial product ions from m/z 533 (the molecular ion).

[0077] Single-crystal X-Ray diffraction analysis was employed to determine the crystal structure of the Form A of ponatinib hydrochloride. Single crystals of ponatinib HCI, in the anhydrate HCI-1 form were obtained using the vapor diffusion into liquid crystallization method using ponatinib HCI CGAM Batch F08-06057. A single crystal obtained using methanol as a solvent with ethyl acetate as anti-solvent was analyzed by single crystal X-ray diffraction. From prior experiments, it was known that crystals of this form diffracted well, leading to the solution of the structure of ponatinib HCI shown in Figure 12, with crystallographic parameters summarized in Table 2. The terminal nitrogen of the piperazine is the site of protonation in ponatinib HCI, consistent with the previously described NMR analysis of ponatinib HCI. The chloride counter-ion occurs in the crystal structure immediately adjacent to the site of protonation. Based on this structure analysis, it was determined that Form A is an anhydrated form.

Table 2: Crystal Data and Structure Refinement for Ponatinib Hydrochloride Form A.

[0078] The attenuated total reflectance (ATR) FT-lR spectrum of Form A of ponatinib HCI, ASI Batch 110020, is shown in Figure 13. Table 3 provides a summary of selected IR band assignment of ponatinib HCI based on the FT-IR shown in Figure 13.

Table 3; Selected IR Band Assignment of Ponatinib HCI

[0079] In the FT-IR, the functional group region extends from 4000 to 1300 cm-1. In the region from 3300 to 2800 cm-1 (region A), there are multiple overlapping, absorption bands arising from stretching vibrations between hydrogen and some other atom, likely amide N-H stretching, aromatic C-H stretching (from the imidazo-pyridazine heterocycle and phenyl groups) and aliphatic C-H stretching (in methyl and methylene groups), all present in the structure of ponatinib HCI. A weak band in the 21OQ-2260cm-1 (region B) is due to triple C-C bond stretching. A medium intensity band due to amide C=0 stretching (Amide 1) can be expected in 1640-1690 cm-1 range, likely the band observed at 1669.8 cm-1(region C). Secondary amide N-H bending gives absorption bands in the 1500-1560cm-1 range (Amide 2), where two strong bands are observed (region D). Multiple weak to medium 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). Aromatic and aliphatic amine C-N stretching bands can be expected in the 1250-1335cm-1 range and in the 1250-1020cm-1 range, respectively, where multiple bands are observed, including a particularly strong band at 1314.9 cm-1 (regions F, G). The fingerprint region, 1300 to 910 cm-1, is complex with a strong, broad band at 1122.6 cm-1 (region H). likely due to C-F stretching. The aromatic region, 910 to 650 cm-1, absorption bands are primarily due to the out-of-plane bending of hetero-aromatic ring C-H bonds indicating the hetero-aromatic nature of the compound (region I). These data in the FT-IR spectrum support the proposed structure of Form A of ponatinib hydrochloride.

[0080] Experiments to determine the purity of Form A were carried out, With reference to Figure 14, it was determined that the purity of Form A of ponatinib hydrochloride is 99,8160 % (area percent).

[0081] Characteristics of Form B fHCI2): [0082] Form HCI2 was obtained from a solubility assessment in TFE/water and it was converted to form HCI2b one day after storage of the measuring plate at ambient conditions, as confirmed by the XRPD re-measurement of the specific sample. HCI2 was also obtained in the experiments performed in Phase 2 described herein from aqueous solvent systems (water and MeOH/waler) and it also converted to form HCI2b upon storage at ambient conditions (see overview in Figure 2).

[0083] Form B was analyzed by X-ray powder diffraction (XRPD). Figure 15 shows XRPD patterns of (from bottom to top): starting material (Form A), Form HCI2 (QSA12.1, solvent: water) and Form HCl2b (QSA12.2, remeasurement after few days at ambient conditions). In the XRPD pattern of Form B shown in Figure 15, at least one or all of the following peaks in degress two theta (2Θ) is shown: 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 of Form C shown in Figure 15, at least one or all of the following peaks in degress two theta (2Θ) is shown: 6.5; 12.4; 13.8; 17.4; 18.0; 20.6; 22.0; 23.0; 25.5; 26.5; and 27.4. In these embodiments, the XRPD pattern of Form B and C shows two peaks, three peaks, four peaks or five peaks.

[0084] Experiments to determine the purity of Form B were carried out. With reference to Figure 15, it was determined that the purity of Form B of ponatinib hydrochloride is 99.7535 % (area percent).

[0085] Characteristics of Form C (HCI2b): [0086] Form B is a hydrated form. Form FICI2b was initially obtained from the solubility assessment experiments, either by conversion of Form B, over a number of days under ambient conditions or directly from TFE/water solvent mixtures, Form C was also obtained in the Phase 2 experiments from aqueous solvent systems (water and water/DMSO) (see overview in Figure 2).

[0087] Form C was analyzed by X-ray powder diffraction (XRPD), Figure 17 shows XRPD patterns of (from bottom to lop): starting material (HCI1) and Form HCI2b (QSA21.1, solvent: water). In the XRPD pattern of Form C shown in Figure 17, at least one or all of the following peaks in degress two theta (2Θ) is shown: 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, the XRPD pattern of Form C shows two peaks, three peaks, four peaks or five peaks, [0088] With reference to Figure 18, the melting point of Form C of ponatinib HCI was determined by differential scanning calorimetry (DSC). The sample of was analyzed in a pin-holed crucible in the temperature range of 25°C to 300°C at a heating rate of 10°C per minute using dry N2 gas purge, Intense endothermic events occurred at TpaS|< = 122.9°C, TpBak =158.2°C and Tpeak =256.2DC.

[0089] Figure 19 shows TGA and SDTGA thermograms of QSA21.1. Figure 20 shows a TGMS thermogram of Form C from experiment QSA21.1. A mass loss of 4.3% (water) is observed in the temperature interval 40°C-140°C. The APkwater ratio was assessed as 1:1.4.

[0090] Experiments to determine the purity of Form C were carried out. With reference to Figure 21, it was determined that the purity of Form C of ponatinlb hydrochloride is 99.7850 % (area percent).

[0091] Characteristics of Form D (HCl3-classh [0092] FiCI3-ciass was mostly obtained from aromatic solvents, as can be seen in the overview in Figure 2, with the exception of the MeOH/acetonitrile mixture. Form D was successfully reproduced at the 120 mg scale using cooling-evaporative crystallization in toluene.

[0093] Based on the thermal analyses, the sample representative of the Form D was assigned as a toluene solvated form (APl.toluene 1:0.5), The form desolvated at 199.5°C, recrystallized and a second melting was observed at 257.6°C (most likely corresponding to the melting point of Form A). HCI3-class is mildly hygroscopic, with 2.5% water mass uptake at 95% RH. The process was reversible regarding to physical stability and sample appearance.

[0094] HCI3-class sample was found to be physically stable after 8 months storage under ambient conditions and following the DVS cycle. Flowever, HCI3-class sample converted to HCI1 after 1 week in the humidity chamber (40°C/75% RFI).

[0095] Form D was analyzed by X-ray powder diffraction (XRPD). Figure 22 shows XRPD patterns of a XRPD overlay of (from bottom to top): HCI1 (AP24534 HC1 salt starting material), FICI3-c|ass (PSM17, solvent: toluene), HCI3 (PSM1, solvent: toluene), HCI1+HCI3 (PSM1 after one week at 40°C/75% RFI) and HCF3 (PSM1 after DVS). In the XRPD pattern shown in Figure 22, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI3: 8.2; 10.1; 10.9; 14.9; 16,0; 16.3; 16.8; 17,7; 18,7; 20.2; 22.9; 24,0; 25.8; 26.7; and 28.5. In the XRPD pattern shown in Figure 22, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI3+HCI1: 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, the XRPD pattern of Form D shows two peaks, three peaks, four peaks or five peaks.

[0096] With reference to Figure 23, the melting point of Form D of ponatinib HCI (PSM1) was determined by differential scanning calorimetry (DSC). The sample of was analyzed in a pin-holed crucible in the temperature range of 25°C to 300°C at a heating rate of 10°C per minute using dry Nz gas purge. Intense endothermic events occurred at Tpeai< = 199,5°C, Tpeak = 204.1DC and TpeaK “ 257.6°C.

[0097] With reference to Figure 24, TGA and SDTGA thermograms of Form D (PSM1) are provided. A mass loss of 7.7% (toluene, ratio APfSolvent is 1:0.51) was observed in the temperature interval 120°C-220°C.

[0098] With reference to Figure 25, a FT-IR spectrum of the region of 1750 - 500 cm'1 is shown, These data support the proposed structure of Form D of ponatinib hydrochloride. In addition, this spectrum show the unique identity of Form D relative to Form A.

[0099] Experiments to determine the purity of Form D (PSM1) were carried out. With reference to Figure 26, it was determined that the purity of Form D of ponatinib hydrochloride is 97.3664 % (area percent).

[00100] Characteristics of Form E (mixture of HCI4+HCI1): [00101] HCI4 was only obtained as a mixture with Form A from a grinding experiment with hexafluorobenzene (see overview in Figure 2).

[00102] Form E of ponatinib hydrochloride was found not to be physically stable upon storage at ambient conditions. The mixture HCI1+HCI4 was re-measured by XRPD after 8 months of storage and it had converted to Form A.

[00103] Characteristics of Form F <HCI5-class): [00104] Form HC!5 was obtained from one vapor diffusion onto solids experiment described here in butyl acetate (see overview in Figure 2). HCl5-class was characterized by DSC, cycling-DSC, TGMS, FTIR, HPLC and DVS. The physical stability under short-term storage conditions (i.e. one week at 40°C and 75% RH) was investigated. HCI5-class samples were physically stable, as assessed by XRPD, after 8 months storage under ambient conditions. After 1 week in the humidity chamber (40°C/75% RH), the material was still HCI5-ciass, however with a slightly different XRPD pattern.

[00105] A DVS experiment showed that HCI5-class is highly hygroscopic, with a 37% water mass adsorption. The material lost its crystallinity as indicated by the XRPD following the DVS experiment.

[00106] Form F was successfully scaled up at the 120 mg scale using the same conditions as those of the original experiment to identify the previously discovered polymorphs. Two scale-up experiments were performed and the corresponding XRPD patterns indicated forms isostructural to HCI5. These isostructural forms together with HCI5 and HCl5b were designated HCI5-class or Form F. Figure 27 shows XRPD patterns of a XRPD overlay of (from bottom to top): HCI1 (Form A starting material); HCI5 and HCI5b (VDS28 wet and dry, solvent: butyl acetate); HCI5-class (VDS1, solvent: butyl acetate), Low crystalline (VDS1 after DVS); HCI5-class (VDS2, solvent: butyl acetate); and HCI5-class (VDS2 after one week at 40°, 75% RH). In the XRPD pattern shown in Figure 27, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI5: 6.8; 9.8; 12.4; 16.2; 17.9; 19.0; 24.0; and 25.1. In the XRPD pattern shown in Figure 27, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI5-class (top pattern): 7.9; 8.7; 9.7; 11.4; 15.6; 16.5; and 25.8. In certain embodiments, the XRPD pattern of Form F shows two peaks, three peaks, four peaks or five peaks.

[00107] With reference to Figure 28, the melting point of Form F of ponatinib HCI (PSM1) was determined by differential scanning calorimetry (DSC). Samples from two different experiments were analyzed in a pin-holed crucible in the temperature range of 25°C to 300“C at a heating rate of 10°C per minute using dry N2 gas purge. A sample from one experiment (VDS1, top curve) evidenced intense endothermic events occurred at Tp<,ak = 120.7°C, Tpeak = 184.3°C and Tpeak = 209.4DG. A sample from another experiment (VDS2, bottom curve) evidenced intense endothermic events occurred at Tpe* = 122.ΓΟ, TPeak = 209.7°C and Tpeak = 252.1°C, [00108] A cycling DSC experiment showed that upon desolvation, HCI5-class converted to a form designated "FICl5-desolvate", which melted at circa 210°C.

[00109] With reference to Figure 29, a TGA/SDTA thermogram of Form F (VDS1, top) and TGMS (bottom) thermogram are provided. A mass loss of 17.1% (Butyl acetate, ratio APLSolvent 1:1,01) was observed in the temperature interval 25°C-160°C. TG-MS analyses showed that HCI5-class is a butyl acetate solvate with a ratio API:butyl acetate of 1:1 and if desolvates at around 120°C. Figure 30 provides a corresponding TGA/SDTA (top) and TGMS (bottom) thermogram of Form F for VDS2. A mass loss of 16.6% (Butyl acetate, ratio APLSoivent 1:0.98) was observed in the temperature interval 25°C-160°C.

[00110] With reference to Figures 31 and 32, a FT-IR spectrum of the region of 1750 -500 cm'1 is shown. These data support the proposed structure of Form F of ponatinib hydrochloride. In addition, these spectra show the unique identity of Form F relative to Form A.

[00111] Experiments to determine the purity of Form F (VDS2) were carried out. With reference to Figure 33, it was determined that the purity of Form D of ponatinib hydrochloride is 98.2833 % (area percent).

[00112] Characteristics of Form G (HCI5b): [00113] Form G or ponatinib hydrochloride was obtained by conversion of HC!5, upon drying for 3 days under full vacuum. HC!5b form was found to be physically stable after 8 months storage under ambient conditions.

[00114] Characterization data for Form G is provided herein in the context of Form F.

[00115] Characteristics of Form H (HCi6-classi: [00116] HCI6 was obtained from two experiments; vapor into solution and vapor onto solids, in MeOH/water and MeOH solvent systems, respectively (see overview in Figure 2). Different time points of material sampling showed that the corresponding XRPD patterns were slightly different, without being bound by theory, indicating that HCI6 is a class of forms, likely isostructural. HCI6-ciass was successfully scaled up to 120 mg using the same conditions as those of the MeOH vapor onto solids experiment of the original screening experiment.

[00117] HCI6-class was characterized by DSC, cycling-DSC, TGMS, FTIR, HPLC and DVS. The physical stability under short-term storage conditions (i.e. one week at 40°C and 75% RH) was investigated. Form H samples were physically stable, as assessed by XRPD, after 8 months storage under ambient conditions. After 1 week in the humidity chamber (40°C/75% RH), the material was still HCI6-class, however with a slightly different XRPD.

[00118] Figure 34 shows XRPD patterns of a XRPD overlay of (from bottom to top): Form A (ponatinib hydrochloride starting material), HCI6-class (VDS6, solvent methanol), HCI6-class (VDS3, solvent: methanol), HCI6 (VDS3 after DVS), HCI6-class (VDS3 after climate chamber), HCI6-class (VDS4, solvent: methano!) and HCI6-class (VDS4 after DVS). In the XRPD pattern shown in Figure 34, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI6 (immediately above 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 the XRPD pattern shown in Figure 34, at least one or all of the following peaks in degress two theta (2Θ) is shown for HCI6-class (top pattern): 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 XRPD pattern of Form F shows two peaks, three peaks, four peaks or five peaks. Although XRPD analysis of both samples showed that similar patterns were observed after the DVS run, the TGMS analysis of VDS4 showed that methanol molecules were no longer present in the sample but they had been replaced by water molecules (forming presumably a hemi-hydrated form belonging to the HCI6~class.

[00119] With reference to Figure 35, the melting point of Form Ft of ponatinib HCI was determined by differential scanning calorimetry (DSC). Samples from two different experiments were analyzed in a pin-holed crucible in the temperature range of 25°C to 300°Ο at a heating rate of 10°C per minute using dry N2 gas purge, A sample from one experiment (VDS3, top curve) evidenced an intense endothermic event at TpBak = 219.4°C. A sample from another experiment (VDS4, bottom curve) evidenced intense endothermic events occurred at Tpeak = 219.40CandTpBBk = 256.8oC.

[00120] With reference to Figure 36, a TGA/SDTA thermogram of Form H (VDS3, top) and TGMS (VDS3, bottom) thermogram are provided. A mass loss of 5,4% (Methanol, ratio API:Solvent 1:1,01) was observed in the temperature interval 30°C -150°C and a mass loss of 0.3% (Methanol ratio APl.Solvent 1:0.05) was observed in the temperature interval 190°C-220°C. A corresponding TGA/SDTA thermogram of Form H (top) and TGMS (bottom) thermogram is shown at Figure 37 for VDS4. A mass loss of 3.3% (Methanol, ratio APl.Solvent 1:0.6) was observed in the temperature interval 30°G-150°C and a mass loss of 0.7% (Methanol, ratio APFSolvent 1:0.12) was observed in the temperature interval 190°C-220°C.

[00121] With reference to Figures 38 and 39, a FT-IR spectrum of the region of 1750 -500 cm"1 is shown. These data support the proposed structure of Form H of ponatinib hydrochloride. In addition, these spectra show the unique identity of Form H relative to Form A.

[00122] Experiments to determine the purity of Form H (VDS4) were carried out. With reference to Figure 40, it was determined that the purity of Form H of ponatinib hydrochloride is 97.9794 % (area percent).

[00123] Characteristics of Form I (HCI6 desolvatel: [00124] A cycling DSC experiment conducted in connection with the experiments for Form H showed that upon desolvation, HCS6-class converted to a form designated “HCI6-desolvate", which melted at circa 22Q°C.

[00125] Characteristics of Form J (HCI71: [00126] Form J is a pentahydrate of ponatinib HCI and was discovered in the context of a single crystal analysis. Form J is the most stable hydrated structure identified, as competitive slurries in water between the trihydrate and pentahydrate showed.

[00127] Single crystals of suitable size were obtained in the vapor diffusion experiment performed with the solvent mixture methanol/water 20:80 and n-butyl acetate as anti-solvent (experiment ID 46). One parallelepiped single crystal of approximate size 0.45 x 0.25 x 0.12 mm was collected from the crystallization vial and mounted on a glass fiber. The crystallographic data (collected up to Θ = 27.5°) are listed in Table 4.

Table 4: Crystal Data and Structure Refinement for Form J

[00128] The asymmetric unit comprises the cation, the chloride anion and five water molecules (pentahydrate). The water molecules are connected via hydrogen bonding (H-Bonds) with the anion, the cation and neighboring water molecules.

[00129] The important consequence of the present Η-Bonds arrangement is the fact that in this crystal both charged atoms (i.e. the protonated nitrogen from the API and the chloride anion) are bridged/separated by several molecules of water.

[00130] Figure 43 shows a characteristic X-Ray Powder Diffraction (XRPD) pattern for Form J of ponatinib hydrochloride. The XRPD pattern of Form J shown in Figure 43 shows at least one or more of the peaks having a relative intensity of 20% or greater in degress two theta (20):. In certain embodiments, the XRPD pattern of Form J shows two peaks, three peaks, four peaks or five peaks.

[00131] Characteristics of Form K (HCl8i: [00132] Form K was discovered in the context of a single crystal analysis. The single crystals were grown in the slow evaporation experiment conducted with TFE/H20 mixture 50:50 (experiment ID 23). One block-like single crystal of approximate size 0.40 x 0.30 x 0.25 mm was analyzed. Although the crystal was large, it diffracted quite poorly, which is an indication of partial disorder in the structure. Therefore the measurement was recorded only up to Θ = 25°. The crystallographic parameters are listed in Table 5,

Table 5: Crystal data and structure refinement for Form K.

[00133] The structure of the mixed TFE solvated/hydrated form comprises the cation, the chloride anion and two neutral entities: the trifluoroethanol and the water molecules. In this structure, although water molecules are involved in the H-bonding they do not separate the charged atoms (contrary to the pentahydrated and trihydrated forms. The TFE and water molecules acted only as donors in the hydrogen bonding network. In particular for the water molecules, only one of the hydrogen atoms acts as donor, which could be responsible for the disorder of the water molecules and the fact that the ratio of the water molecules compared to the API molecules is not stoichiometric.

[00134] Figure 44 shows a characteristic X-Ray Powder Diffraction (XRPD) pattern for Form K of ponatinib hydrochloride. The XRPD pattern of Form K shown in Figure 44 shows at least one or more of the peaks having a relative intensity of 20% or greater in degress two theta (2Θ):. In certain embodiments, the XRPD pattern of Form K shows two peaks, three peaks, four peaks or five peaks.

[00135] Characteristics of amorphous form of ponatinib hydrochloride: [00136] Figure 45 shows XRPD patterns of Form A of ponatinib hydrochloride (bottom pattern) and amorphous ponatinib hydrochloride (top pattern) (solvents,2,2-trifluroethanol). It is readily apparent that Form A has a distinct set of peaks at particular angles two theta whereas the amorphous ponatinib hydrochloride does not have any defined peaks.

[00137] In addition, amorphous ponatinib hydrochloride has a unique melting temperature as compared to Form A of amorphous ponatinib hydrochloride. Figure 46 shows a characteristic Differential Scanning Calorimetry (DSC) thermogram of amorphous ponatinib hydrochloride. An intense endothermic event with a peak of 259.4°C was observed, corresponding to the melting point of the amorphous form. This melting point is distinct from that observed with Form A of ponatinib hydrochloride, which demonstrated a melting point of 264.1°C.

[00138] Unique and distinct physical properties of amorphous ponatinib hydrochloride and Form A of ponatinib hydrochloride do not seem to be attributed to purity of the respective materials. In the case of amorphous ponatinib hydrochloride, the material was determined by HPLC to have a purity of 99.7877% (area percent) (see Figure 47), whereas the purity of Form A of ponatinib hydrochloride was determined to be 99.8% (area percent). EXAMPLE 1

DISCOVERY OF POLYMORPHIC FORMS

[00139] Initial efforts to discover polymorphic forms of ponatinib hydrochloride were divided into two phases. Phase 1 included starting-material characterization, feasibility testing and solubility study to provide data for the solvent selection for Phase 2. Phase 2 included 192 polymorph screening experiments at milliliter (ml) scale. These initial efforts led to the discovery of eight polymorphic forms, Form A, Form B, Form C, Form D, Form E, Form F, Form G and Form H.

[00140] Phase 1: Starting Material Characterization [00141] Approximately 24 grams of the compound ponatinib hydrochloride was provided as a light 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 mono hydrochloride, is provided as a crystalline material (designated HCI1) and its chemical purity was assessed by HPLC as 99.8%. TGA and TGMS analyses showed 0.7% of mass loss (residua! ethanol) in the temperature interval 25°C-240°C prior to the thermal decomposition process. DSC analysis showed an endothermic event with Tpa0k = 264,8*0, probably related to melting and/or decomposition of the compound, 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-y!)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride.

[00142] Phase 1: Solubility Study [00143] Quantitative solubility testing was performed on ponatinib hydrochloride starting material, employing a set of 20 solvents. Slurries were prepared with an equilibration time of 24 hours after which the slurries were filtrated. The solubility was determined from the saturated solutions by HPLC. The residual solids were characterized by XRPD. The results are summarized in Table 6.

Table 6: Solubility Study of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methy1]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride

All tests were conducted at room temperature with stirring. 1 The solid form obtained from tho slurry was assessed based on the XRPD analysis. 1 Under Range, lower thBn detection limit, the concentration is lower than 0.22 mg/ml 3 Over Range, the material was dissolved, the concentration is higher then 200 mg/ml. ' Amorphous [00144] In 19 of the experiments shown in Table 6, the materials analyzed following the solubility assessments in 19 different solvents appeared to be the same form as the starting material designated form HCI1. In the experiment QSA13 performed in 2,2,2-trifluoroethanol. the material dissolved completely at the selected concentration and the sample obtained after evaporation of the solvent resulted in amorphous material. The solids from two slurries in water (QSA12 and QSA21) resulted in two different forms, Form HCI2 and Form HCI2b, respectively. After few days stored at ambient conditions, the form HCI2 converted to Form HC!2b and it could therefore not be further characterized. Upon further characterization, the Form HC!2b was determined to be a hydrated form (ratio API/water 1:1.4).

[00145] Phase 1: Feasibility Study [00146] Feasibility tests were performed to attempt to obtain amorphous starting material that could be employed in some crystallization techniques of the Phase 2 portion of the study. Two techniques were employed i.e. grinding and freeze-drying. The results are presented below.

[00147] Grinding. Two grinding experiments were performed with two different durations at a frequency of 30Flz. After 60 minutes of grinding, the crystalline starting material converted to amorphous. After 120 min, the resulting material remained amorphous with a chemical purity of 99.6%.

[00148] Freeze-drying. Eight freeze-drying 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 mono hydrochloride. These experiments are summarized in Table 7.

Table 7: Freeze-drying feasibility study of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1 -yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride

1 Based on the TGMS results. 3 Chemical purity determined by HPLC.

[00149] The solubility of compound ponatinib hydrochloride in tetrahydrofuran, 2-methyltetrahydrofuran and dichloromelhane was too low to apply the freeze drying procedure in good conditions. With solvents such as methanol, 2,2,2-trifluoroethanol (TFE) and TFE/water mixtures, amorphous material was obtained. In the samples obtained from neat TFE or with high TFE content in the solvent mixtures, 11% of residual solvent was detected in the dried powders (according to the TGMS results). The samples obtained from methanol and TFE/water 50:50 contained less residual solvent only 0.9% and 1.5%, respectively. The amount of residual solvent in the amorphous material produced from TFE/water 50:50 could be reduced to below 1% after extra drying for 24 hours. For both amorphous samples obtained from methanol and TFE/water 50:50, the chemical purity was assessed to be 99.8% by FIPLC. Because creeping was observed in the freeze-drying experiment with methanol, the procedure using TFE/water 50:50 was selected to be used to produce the amorphous ponatinib hydrochloride to be used in the cooting-evaporation crystallizations and vapor diffusion onto solids experiments of Phase 2, [00150] Phase 2: Polymorph Discovery [00151] The polymorph screening experiments for ponatinib hydrochloride were carried out at milliliter (ml) scale using 192 different conditions in which six different crystallization procedures were applied: (1) cooling-evaporation; (2) anti-solvent addition; (3) grinding; (4) slurry; (5) vapor diffusion into solutions; and (6) vapor diffusion onto solids. After the screening experiments were completed, the materials were collected and analyzed by XRPD and digital imaging.

[00152] Cooling-Evaporative Crystallization Experiments. The 36 cooling-evaporative experiments shown at Table 8 at ml scale were performed in 1.8 ml vials, employing 36 different solvents and solvent mixtures and 1 concentration. In each vial, 25 mg of amorphous ponatinib hydrochloride was weighed. Then the screening solvent was added to reach a concentration of circa 60 mg/ml. The vials, also containing a magnetic stirring bar, were closed and placed in an Avantium Crystal16 to undergo a temperature profile as described in Table V. The mixtures were cooled to 5°C and held at that temperature for 48 hours before placing the vials under vacuum. The solvents were evaporated for several days at 200 mbar or 10 mbar and analyzed by XRPD and digital imaging.

Table 8: Experimental conditions for the 36 ml experiments using the cooling-evaporation method.

Table 9: Temperature profile employed for the 36 cooling-evaporative experiments

[00153] Crash-crystallization with anti-solvent addition Experiments. Eor the crash-crystallization experiments. 36 different crystallization conditions were applied, using 1 solvent and 24 different anti-solvents (see Table 9). The anti-solvent addition experiments have been performed forwards. A stock solution was prepared, the concentration of ponatinib hydrochloride being that attained at saturation at ambient temperature after equilibration for 24 hours before filtering info 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. Because no precipitation occurred, this ratio was increased to 1:4 with a waiting time of 60 minutes between each addition. As no precipitation occurred yet, the solvents were completefy evaporated under vacuum at room temperature. After evaporation, the experiments resulted to have no yield.

Table 9: crash-crystallization experiments

[00154] Grinding Experiments. The drop-grinding technique uses a small amount of solvent added to the material 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-j(4-methylpiperazm-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride, which is grinded in a stainless steel grinder jar with 2 stainless steel grinding bails. In this manner, the effect of 24 different solvents (see Table 10) was investigated. Typically 30 mg of starting material was weighed in the grinding container and 10 μ| of solvent was added to the container. The grinding experiments were performed at 30 Hz during 120 min. Each wet material was subsequently analyzed by XRPD and digital imaging.

Table 10: Experimental Conditions for Grinding Experiments.

_______________

[00155] Slurry Experiments. A total of 48 slurry experiments were performed with the compound ponatinib hydrochloride and 24 solvents at 10°C and 30°C, for 2 weeks. Table 11 summarizes the experimental conditions. The experiments were carried out by stirring a suspension of the material In a solvent at a controlled temperature. At the end of the slurry time, the vials were centrifuged and solids and mother liquids separated. The solids were further dried under full vacuum at room temperature and analyzed by XRPD and digital imaging.

Table 11: Experimental Conditions for the Slurry Experiments

.

[00156] Vapor Diffusion Into Solutions. For the vapour diffusion experiments, saturated solutions of ponatinib hydrochloride were exposed to solvent vapours at room temperature for two weeks. A volume of saturated solution was transferred to an 8 ml vial which was left open and placed in a closed 40 ml vial with 2 ml of anti-solvent (see Table 12). After two weeks, the samples were checked for solid formation. The samples were dried under vacuum (200 mbar or 10 mbar) and resulted to have no yield. Based on the results, additional experiments were performed with 12 different crystallization conditions as described in the table, experiments ID VDL25 - VDL3B.

Table12: Experimental Conditions for the Vapor Diffusion Into Solutions.

[00157] Vapor Diffusion Onto Solids. For the vapour diffusion experiments, amorphous ponatinib hydrochloride was exposed to solvent vapours at room temperature for two weeks.

The 8 ml vials with the amorphous API were left open and placed in a closed 40 ml via! with 2 ml of anti-solvent (see Table 13). After two weeks, the solids were analyzed by XRPD and digital imaging, if the solids were liquefied by the vapours, the samples were dried under vacuum (200 mbar or 10 mbar) before they were analyzed by XRPD and digital imaging.

Table 13: Experimental Conditions for the Vapor Diffusion Onto Solids.

[00158] Analytical Methods Applicable for Phase 1 and Phase 2 Experiments: [00159] X-ray powder diffraction [00160] Following the evaporation experiments, the products were harvested. XRPD patterns were obtained using an Avantium T2 high-throughput XRPD set-up, The plates were mounted on a Bruker GADDS diffractometer equipped with a Hi-Star area detector. The XRPD platform was calibrated using Silver Behenate for the long d-spacings and Corundum for the short d-spacings.

[00161] Data collection was carried out at room temperature using monochromatic CuKa radiation in the 2Θ region between 1.5° and 41.5°, which is the most distinctive part of the XRPD pattern. The diffraction pattern of each well was collected in two 2Θ ranges (1.5 °< 2Θ < 21.5 0 for the first frame, and 19.5 °< 26 £ 41.5 ° for the second) with an exposure time of 90 seconds for each frame. No background subtraction or curve smoothing was applied to the XRPD patterns. The carrier material used during XRPD analysis was transparent to X-rays and contributed only slightly to the background.

[00162] Thermal Analysis [00163] Melting properties were obtained from DSC thermograms, recorded with a heat flux DSC822e instrument (Mettler-Toledo GmbH, Switzerland). The DSC822e was calibrated for temperature and enthalpy with a small piece of indium (m.p. = 156.6°C; AHf - 28.45 J.g'1). Samples were sealed in standard 40 pi aluminum pans, pin-holed and heated in the DSC from 25°C to 300°C, at a heating rate of 10°C min'1. Dry N2 gas, at a flow rate of 50 ml min'1 was used to purge the DSC equipment during measurement.

[00164] Mass loss due to solvent or water loss from the crystals was determined by TGA/SDTA. Monitoring the sample weight, during heating in a TGA/SDTA851e instrument (Mettler-Toledo GmbH, Switzerland), resulted in a weight vs. temperature curve. The TGA/SDTA851e was calibrated for temperature with indium and aluminum. Samples were weighed into 100 μΙ aluminum crucibles and sealed. The seals were pin-holed and the crucibles heated in the TGA from 25 to 300°C at a heating rate of 10°C min'1. Dry N2 gas was used for purging.

[00165] The gases evolved from the TGA samples were analyzed by a mass spectrometer Omnistar GSD 301 T2 (Pfeiffer Vacuum GmbH, Germany). The latter is a quadruple mass spectrometer which analyses masses in the range of 0-200 amu.

[00166] Digital Imaging [00167] Digital images were automatically collected for all the wells of each well-plate, employing a Philips PCVC 840K CCD camera controlled by Avantium Photoslider software.

[00168] Press [00169] For the compression tests, an Atlas Power Press T25 (Specac) was used. The

Atlas Power T25 is a power assisted hydraulic press operating up to 25 Tons.

[00170] HPLC Analytical Method [00171] HPLC analysis was performed using an Agilent 120OSL HPLC system equipped with UV and MS detectors following the conditions presented below:

HPLC Equipment: LC-MS

Manufacturer: Agilent HPLC: HP1200sl

UV-detector: HP DAD MS-detector: HP1100 API-ES MSD VL-type

Column: Waters Sunfire C18 f 100 x 4.6mm: 3.Sum).

Column temp: 35 °C

Mobile phase: Gradient mode

Mobile phase A: 1000/1; H20/TFA (v/v)

Mobile phase B: 1000/1; ACN/TFA (v/v)

Flow: 1.0ml/min

Gradient program: Time [min]: % A: % B: 0 90 10 15 20 80 16 90 10 18 90 10

Posttime: 1

UV-Detector: DAD 200-400 nm

Wavelength: 260 nm 4 nm

Time: 0-17 min

MS-Detector: MSP

Scan: positive

Mass Range: 70 - 1000 amu

Fragmentator: 70

Time: 0-17 min

Autosampler:

Not controlled

Injection mode: loop

Injection volume: 5 pi

Needle wash: 2/3; ACN/H20 (v/v)

Dilution solvent: 2,2,2-Trifluoroethanol

The compound integrity is expressed as a peak-area percentage, calculated from the area of each peak in the chromatogram, except the ‘injection peak’, and the total peak-area, as follows:

The peak-area percentage of the compound of interest is employed as an indication of the purity of the component in the sample.

[00172] In the crystallization experiments during these initial efforts, XRPD analysis of the dry (and if applicable wet) samples obtained revealed the presence of seven additional polymorphic forms in addition to amorphous maleriais and the starting material, Form A. The seven forms are designated HCI2, HCI2b, HCI3-class, HCI5, HCL5b, HCI6-class and the mixture HC11+HCI4, [00173] The occurrence of the different forms obtained in Phase 2 of these initial efforts is presented in Figure 2. XRPD patterns and digital images representative of each form obtained in these Phase 2 experiments were obtained. The characterization of the forms obtained in Phase 1 of these initial efforts is summarized in Table 14,

Table 14: Characterization of Certain Polymorphic Forms of ponatinib hydrochloride

Occ: the total occurrence included 216 experiments carried out in Phase 2 for which 39 samples were analyzed additionally wel or the mother liquor was evaporated and analyzed giving a total of 254 materials characterized. For example, "(3, 1.2%)" correspond to 3 occurrences of the form out of 254 measurements, giving a percentage of 1.2%. For 62 out of the 254 measurements (9%), the product yield or the scattering intensity of some products was too low to identify the solid form, or the materials were wet. b Crystallization modes: cooling-evaporative (PSM), crash crystallization with anti-solvent addition (AS), grinding (GRP), slurry (SLP), vapour diffusion onto solid (VDS) and vapour diffusion into solution (VD1.). Freeze-drying (FD) was used to produce amorphous material (see Phase I experiments). QSA (quantitative solubility experiment), see Phase 1 experiments. c Solvation slate assessed from the TGMS results. 11 Endotherms assessed from the DSC results. ' Chemical purity assessed from HPLC results. rNot determined in this experiment. E Structure determined by single crystal analysis.

[00174] The polymorphic forms identified in these Phase 1 and Phase 2 experiments and shown in Figure 2 were assigned primarily on XRPD analysis. In the course of this analysis, it was observed that some patterns had similarities in the general fingerprint of the XRPD pattern but showed some small differences like peaks shifting or smaller additional peaks. These types of patterns were clustered as a class of patterns (e.g. HCI3-Class). Based on the XRPD, it was concluded that the similarity between the XRPD patterns within a class is explained by the fact that these solid forms are isomorphic hydrates/solvates (similar crystal packing but slightly different unit cell parameters caused by the incorporation of the different solvents and water in the crystal structure).

[00175] The classes of isomorphic solvates were designated by a number (HCI3-Class) or a number-letter combination (for example HCI2 and HCI2b). The class of isomorphic solvates/hydrates designated by a letter-number combination indicates that few sub-classes were observed for this class in the experiment (example HCI2 and HCI2b). When more than three sub-classes could be identified within the class, all XRPD patterns corresponding to a class of isomorphic solvates/hydrates were regrouped under one number (example HCI3-Class}.

[00176] The isomorphic solvates within a certain class or between classes designated with the same number showed a higher degree of similarity of their XRPD patterns than in the case of the classes of isomorphic solvates designated with different numbers. For these different classes of isomorphic hydrates/solvates, the larger differences in the XRPD patterns reflect that the crystal structure packing is significantly different.

[00177] In some XRPD patterns, one or two additional peaks were observed compared to the identified forms. Since these peaks could not be assigned clearly to the known forms, they were indicated as "plus peaks". EXAMPLE 2

FURTHER DISCOVERY OF POLYMORPHIC FORMS

[00178] Follow on efforts were undertaken to analyze single crystals of 3-(imidazo[1,2-b]pyridazinT5-ylethynyl)-4~methyl-N-{4-[(4-rnethylpiperazin-1-yl)rnethyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride. Such efforts led to the discovery of five different pseudo polymorphs with two of these additional polymorphic forms being previously undiscovered. These two newly discovered polymorphic forms are designated herein as HCI7 (also referred to herein as “Form J”) and HCI 8 (also referred to herein as "Form K"). In these later experiments, three different crystallization techniques were used to grow single crystals of suitable size for analysis: (1) slow evaporation of crystallization solvent; (2) diffusion of antisolvent into a solution of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride; and (3) temperature controlled crystallization. In total, 54 crystallization experiments were performed in these later experiments to attempt to grow single crystals of the hydrated form of ponatinib hydrochloride salt for structure determination.

[00179] With regard to temperature controlled crystallization, 24 experiments were prepared with mixtures of alcohols and water (see Table 15). For each experiment, 10 mg of ponatinib hydrochloride was used. The mixtures of API and solvents were heated fast up to 80°C and slowly cooled to room temperature (0.1°C/min).

Table 15: Experimental Conditions of the Temperature Controlled Crystallization Experimental Exp Alcohol [μΐ] Water [μΙ] Water/Alcohol Outcome

[00180] With regard to vapor diffusion into solution, 25 experiments were performed, For each experiment, 10 mg of ponatinib hydrochloride was dissolved in 1 ml of mixture of TFE/Water (10:90) or MeOH/Water (30:70). Each solution was placed in a 6 mi vial, which was inserted in a 20 ml vial containing 3 ml of anti-solvent. The vials were kept at room temperature for 2-4 weeks. The details are reported in Table 16,

Table 16: Experimental Conditions of the Vapor Diffusion Experiments Exp Solvent Anti-solvent Outcome

[00181] With regard to slow evaporation of solvents, 10 mg of ponatlnib hydrochloride was placed in an 8 ml vial and 2 ml of solvent (mixture of solvents) was added. In those cases in which the solids did not dissolve, the vial was heated to 90°C. Subsequently, the mixture was left to cool slowly to room temperature (see Table 17).

Table 17: Experimental conditions of the slow evaporation experiments Exp Solvents (ratio) Temperature [,JC] Outcome

[00182] Each of the polymorphic forms disclosed herein are made from specific crystailization/solvent modes using ponatinib HCI as the starting material. While the synthesis of ponatinib HCI has been described previously (e.g., WO 2007/075869 and WO 2011/053938), the following synthesis of ponatinib HC! is provided. EXAMPLE 3

STRESS TEST OF FORM A

[00183] Form A is a crystalline, anhydrous solid that has been reproducibly obtained from a range of solvents. Form HCI-1 is intrinsically chemically stable, which directly correlates to the thermodynamic stability of the HCI 1 form. Form HCI-1 is stable to thermal, pressure, and humidity stress as well as exposure to some solvent vapors, and is thermodynamically stable, * The material had not completely dissolved after having been for several hours at this temperature

Numerous studies have been conducted to confirm its stability in both the formulated (tablets) and unformulated (drug substance) state. The results of such studies are provided in Table 18 below:

Table 18: Stress Studies on Form A

Form HCI-1 is stable to thermal, pressure, and humidity stress as well as exposure to some solvent vapors, and is the most thermodynamically stable solid form isolated to date.

Experiments were carried out to test the physical stability of the crystalline Form HCI1 as follows: - The crystalline Form HCI1 and a physical mixture of HCI1 and amorphous material 50:50 were exposed to ethanol vapour for two weeks (see vapour diffusion experiments) - Tablets were prepared by subjecting the crystalline Form HCI1 to a pressure of 50 and 100 kN/cm2 (or 4 and 8 ton/cm2) for 10 sec.

Form A was stored in capsules for up to 17 months at ambient conditions. These samples were analyzed by high resolution XRPD.

The results obtained are summarized in Table 19 and the XRPD measurements and digital images were obtained. They showed that within the stress conditions applied, the polymorphic Form HCI1 remained unchanged, confirming its good physical stability.

Table 19. Results of follow-up work on Form A

EXAMPLE 4

STABILITY OF CERTAIN POLYMORPHIC FORMS

[00184] Samples of the 8 solid forms of HCI salt were chosen to study their physical stability. Two samples representative of each relevant polymorphic forms of the HC! salt obtained were selected. Each sample was re-analyzed by XRPD. The physical stability of the forms after being stored at ambient conditions for 8 months. The results are summarized below: • HCI1, HCI2b, HCI3-Class, HCI5b and HCI6-Class are physically stable under the investigated conditions; • HCI2 converted to HCI2b (this conversion already occurred after storage of the sample under ambient for 1 day); • HCI5 converted to HCI5b (this conversion already occurred following drying for 3 days under full vacuum); • The mixture HCI1+HCI4 converted to HCI1 after 8 months at ambienl conditions.

Table 20. Physical stability of forms of HCI salt.

In grey shading, results obtained from prior polymorph discovery efforts. “ Starting material (SM): Form HCI I or amorphous material (Am) obtained by freeze-drying. bAs classified by XRPD alter completion of the crystallization experiment. ' Crystallization modes: cooling-evaporative (PSM), crash crystallization with anti-solvent addition (AS), grinding (GRP), slurry (SLP), vapour diffusion onto solid (VDS) and vapour diffusion inlo solution (VDL). QSA (quantitative solubility experiment), 11 Solvation state assessed from the TGMS results. ' Nd=not determined. F HC12 and HC15 converted to HCI2b and HCISb after respectively storage at room temperature for a I day or drying under vacuom for 3 days. EXAMPLE 5

PREPARATION OF FORM A

[00185] Form A of ponatinib HCI is formed as a crystalline material by addition of a solution of HCI (1.0 equivalents) in ethanol to an ethanolic solution of the ponatinib free base. The drug substance, ponatinib HCI, is crystallized in the last step of the drug substance synthetic process by addition of seed crystals which results in a very consistent and characteristic particle size and range for the drug substance. Ethanol content in the last 10 multi-kilogram scale batches of ponatinib HCI in the HCI-1 form ranged from 0.8-1.2%.

[00186] No evidence of ethanol or water was found in the HCI-1 form; hence Ihe Form A is an anhydrate. In addition, the crystal packing of the HCl-1 form does not contain voids capable of accommodating ethanol or other small organic molecules. Additional studies to investigate the ethanol content and the removal of ethanol from ponatinib HCI during drying have indicated that the ethanol appears to be associated with the surface of the crystals in Form A of ponatinib HCI.

[00187] Form HCl-1 is characterized by the consistent presence of residual ethanol in all batches of drug substance at a level of approximately 1% by weight. Crystallographic studies and other studies have shown that residual ethanol is present (trapped) on the surface of the crystals, and is not part of the crystalline unit cell, and that HCl-1 is not an ethanol solvate or channel solvate. Ethanol levels in the last ten multi-kilogram scale drug substance batches have ranged from 0.8 to 1.2%. EXAMPLE OF 6

SYNTHESIS OF PONATINIB HYDROCHLORIDE

[00188] Ponatinib HCI is the product of the convergent four step synthesis depicted in Scheme 1. Step 1 involves the synthesis of the "methyl ester” intermediate AP25047 from starting materials AP24595, AP28141, and AP25570. Step 2 involves the synthesis of the "aniline" intermediate, AP24592, from starting material AP29089. Step 3 is the base catalyzed coupling of AP25Q47 and AP24592 to generate ponatinib free base, also designated as AP24534, which is isolated as the free base. Step 4 is the formation and crystallization of the mono-hydrochloride salt of ponatinib in ethanol.

[00189] A preferred route of synthesis of ponatinib HCI is designated as Process C. Scheme 1: Process C

[00190] Step 1: Synthesis of AP25047 (“Methyl Ester") Intermediate [00191] Step 1 of the ponatinib HCI process is the synthesis of the methyl ester intermediate AP25047 in a three reaction sequence (designated 1a, 1b, and 1c), carried out without intermediate isolation (“telescoped"), from starting materials AP24595, AP25570, and AP28141, as depicted in Scheme 2. The array of two aromatic ring systems connected by a single alkyne linker is constructed through 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 catalysts and process by-products. These operations include the crystallization of AP25047 as the HCI salt from a non-polar solvent, toluene (Unit Operation 1,3), an aqueous work-up and silica gel plug filtration (Unit Operation 1.4), and crystallization from a polar solvent, 2-propanol (Unit Operation 1.5). The two crystallizations provide orthogonal purifications for rejection of related substance impurities with differing polarities. The crystallization and solvent wash of the HCI salt from toluene is controlled by an in-process analytical test for a specific process impurity. The final crystallization of the AP25047 intermediate from 2-propanol has been subjected to multi-variate DoE studies to define the design space for robust rejection of other impurities arising from the telescoped reactions. A series of eight in-process tests in Step 1 provide quantitative, analytical control for reaction completions, impurity rejection, and effective removal of residual solvents.

Scheme 2: Step 1 - Synthesis of AP25047

1st Sonogashira Reaction AP24595, palladium tetrakis triphenylphosphine {PdfPPhj),)), copper (I) iodide (Cut), triethylamine, and tetrahydrofuran (THF) are charged to the reactor. The mixture is stirred and degassed with nitrogen and then pre-degassed AP28141 is charged. The resulting mixture is brought to 45 - 55°C and held for not less than 3 hours, The reaction completion is determined by IPC-1 (HPLC). If the IPC-1 criterion is met, the mixture is concentrated to a target volume and cooled.

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

Unit Operation 1.3: Isolation of AP25047-HCI

While stirring, the cool reaction mixture is quenched by addition of hydrogen chloride gas. A precipitate forms, and residual hydrogen chloride is removed from the suspension by a nitrogen purge. Tetrahydrofuran (THF) is replaced with toluene by an azeotropic distillation under reduced pressure. The resulting warm slurry is filtered in an agitated filter dryer and the fitter cake is triturated and washed with warm toluene. The content of process impurity AP29116 is determined by IPC-4 (HPLC). If the IPC-4 criterion is met, the wet filter cake is dried with agitation under a flow of nitrogen and reduced pressure at 35 - 45°C (jacket temperature). The drying is monitored by IPG-5 (LOD, gravimetric). If the IPC-5 criterion is met, the crude AP25047 HCi is discharged and packaged in FEP bags in a plastic container. The isolated AP25047 HCI can be held for up to 7 days prior to forward processing. U nit Operation 1.4: Work-up

The crude AP25047 HCI solid is charged to a reactor with dichloromethane (DCM) and washed with aqueous ammonia. The aqueous phase is back extracted with DCM for yield recovery purposes and the combined organic phase is 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 strips). If the IPC-6 criterion is met, the organic phase is treated with aqueous sodium bicarbonate until the aqueous wash reaches a pH of NLT 7, as indicated by IPC-7 (pH strips). The organic phase is briefly concentrated followed by the addition of fresh dichloromethane. The organic solution is passed through a silica gel pad, which is then rinsed with additional fresh dichloromethane for increased product recovery.

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 targeted final volume range. The resulting suspension is then cooled and further aged with agitation.

Unit Operation 1.6: Isolation / Drying

The precipitated product is isolated in an agitated filter dryer under a flow of nitrogen, and the filter cake is rinsed with 2-propanol. The wet filter cake is dried with agitation under a flow of nitrogen and reduced pressure at 45 - 55°C (jacket temperature). The drying is monitored by I PC-8 (LOD, gravimetric). If the IPC-8 criterion is met, the product is sampled and packaged into polyethylene bags and placed within a heat sealed mylar coated aluminum foil bag, within an HDPE shipping container (Expected yield range, 65 - 89%).

Step 2: Synthesis of AP24592 (“Aniline") Intermediate Summary and Synthetic Scheme

Step 2 of the ponatinib HCI process is the synthesis of the aniline intermediate, AP24592, by catalytic hydrogenation of the nitro-aromatlc starting material AP29089, as depicted below. The reaction is carried out in ethyl acetate, a solvent in which the starting material 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. At the completion of the reaction, a solvent exchange from ethyl acetate to π-heptane via distillation prompts the spontaneous crystallization of AP24592, resulting in material with high purity. This crystallization has been shown to have a significant purification effect, as most of the process impurities remain solubilized in n-heptane. The three in-process controls in Step 2 are an HPLC of the reaction mixture to confirm consumption of starting material, a GC measurement of ethyl acetate following the azeotropic solvent exchange to π-heptane, and a gravimetric determination of solvent loss on drying.

Step 2: Synthesis of AP24592

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

Unit Operation 2.2: Hydrogenation

The reactor is pressurized with hydrogen until a stable pressure range is achieved and the mixture is then stirred under hydrogen atmosphere for at least 4 additional hours. The reactor is depressurized and a sample taken to assess reaction completion (1PC-1). If the IPG-1 criterion is met, the process is continued to Unit Operation 2.3

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 additional ethyl acetate. The combined filtrate and wash solution is concentrated under vacuum to remove a target volume of ethyl acetate. n-Heptane is charged, and the distillation is continued under vacuum to a target volume. The ethyl acetate content is determined by IPC-2 (GC). If the I PC-2 criterion is met, the process is continued to Unit Operation 2.4.

Unit Operation 2.4: Isolation I Drying

The solid product is dried under vacuum at a target temperature range. The end of drying is determined by IPC-3 (LOD, gravimetric). AP24592 is obtained as a white to yellow solid in a range of 80 - 97 % (based on AP29089 input).

Step 3: Synthesis of Ponatinib Free Base Summary and Synthetic Scheme

Step 3 is the synthesis of the free base of ponatinib by the base-catalyzed reaction of AP25047 and AP24592, presented in Scheme 3. The reaction is carried out in the presence of a strong base, potassium ferf-butoxide, under essentially water-free conditions to minimize the undesired hydrolysis of the methyl ester of AP25D47 to the corresponding unreactive carboxylic acid, AP24600. The presence of this by-product results in not only loss of yield, but in complications in downstream processing during the reaction workup. Drying of the reaction mixture by a series of azeotropic distillations, controlled by an in-process test for water, ensures a robust reaction and nearly quantitative consumption of starting materials. The parameters of the reaction conditions and crystallization, in which process impurities are robustly rejected, are well understood on the basis of DoE studies.

Scheme 3: Step 3 - Synthesis of AP24534 Free Base

Unit Operation 3.1: Drying Reaction Mixture AP25047, AP24592, and 2-methyl tetrahydrofuran (2-Me-THF) are charged to a reactor. The mixture is concentrated at reduced pressure to a target volume. Additional 2-methyl tetrahdyrofuran is added and the distillation repeated. Following another charge of 2-methyl tetrahydrofuran and a distillation cycle, the water content of the mixture is determined in IPC-1(KF). If the IPC-1 criterion is met, the process is continued to Unit Operation 3.2.

Unit Operation 3.2: Reaction

The suspension is maintained with stirring at a target temperature of 13 - 23°C range while potassium terf-butoxide (KOfBu) is charged. After a period of not less than 3 hours, the reaction progress is determined by HPLC (IPC-2). If the IPC criterion is met, the process is continued to Unit Operation 3.3.

Unit Operation 3.3: Quench and Extractions

The reaction mixture is diluted with 2-methyltetrahydrofuran (2-Me-THF), and quenched by the addition of aqueous sodium chloride solution. The organic layer is separated and the aqueous iayer is extracted twice with 2-methyl tetrahydrofuran. The combined organic layers are sequentially washed with aqueous sodium chloride and water. The organic layer is then aged at 15- 30°C.

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 a target volume. 1-Propanol is charged and allowed to stir at elevated temperature to furnish a solution, which is distilled under vacuum to a target volume and then cooled slowly to a temperature range of 20 - 30°C.

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 charged to the suspension with stirring and the resulting suspension is aged for an additional 60 - 120 minutes at 20 - 30°C with agitation prior to isolation in the next Unit Operation.

Unit Operation 3.6: Isolation /Drying

The slurry generated 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 the IPC criterion is met, the product is discharged as an off-white to yellow solid and packaged in double polyethylene bags for storage at ambient temperatures.

Step 4: Synthesis of Ponatinib HCI Summary and Synthetic Scheme

Step 4 of the ponatinib HCI process is the formation of the mono-hydrochloride salt through combination of equimolar quantities of ponatinib free base with hydrochloric acid in ethanol and induction of crystallization through seeding. The parameters of this process have been examined in DoE studies for effects on the generation of the desired solid form and particle size distribution of this process.

The synthetic scheme for Step 4 is presented in Scheme 4Error! Reference source not found..

Scheme 4: Step 4 - Synthesis of Ponatinib HCI

Unit Operation 4.1: Dissolution AP24534 free base and absolute ethanol (EtOH) are charged to a reactor and stirred at 60 -75°C to generate a solution. Dissolution is verified by visual observation.

Unit Operation 4.2: Clarification

The solution is passed through a filter, which is then washed with ethanol at 60-78°C.

Unit Operation 4.3: Acidification / Seeding

The product solution is concentrated under vacuum to a target volume. With stirring, an initial portion (approximately 25%} of a solution of 1N hydrogen chloride in ethanol is then charged to the reactor. The solution is treated with qualified seed crystals of AP24534 HCI at a temperature of 60-70°C to initiate crystallization. The process is continued to Unit Operation 4.4.

Unit Operation 4.4: Crystallization

Once the presence of solids in the reactor is verified by visual observation, the remainder (approximately 75%) of the 1N hydrogen chloride solution in ethanol is slowly added to the stirred mixture. The mixture is aged for at least 10 minutes and IPC-1 is performed to determine the pH of the solution. If the I PC criterion is met, the mixture is cooled to a temperature of 5* 15°C and aged with stirring.

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 product by slow agitation and nitrogen flow at ambient temperature. The solid is then dried under vacuum at 60-70cC. The drying is monitored by I PC-2 (LOD, gravimetric). If the IPC-2 criterion is met, ponatinib HCI is discharged as an off-white to yellow solid and packaged in double polyethylene bags for storage in plastic drums at 20 - 30“C.

Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavour to which this specification relates.

Claims (43)

1. THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

   1. Crystalline 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methy!p!perazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide hydrochloride.
2. 2. Substantially pure crystalline 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1“yl)methyl]-3-(trifluoromethyl)phenyl}benzamide hydrochloride of claim 1.
3. 3. Crystalline Form A of 3-{imidazo[1,2-b]pyridazin-3-ylethynyl)-4-rr>ethyl-N-{4-[(4-methylpiperazin-1 -yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride.
4. 4. The crystalline Form A of claim 3, wherein thecrystaHineFormAof3-(imidazo[1,2-b]pyridaziri-3-ylethynyl)-4-methy!-N-{4-[(4-methylpiperazin-1-yl)methy!]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride Is substantially pure.
5. 5. The crystalline Form A of claims 3 or 4 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 4.
6. 6. The crystalline Form A of claims 3 or 4 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about 5.9; 7.1; 10.0; 12.5; 16.4; 19.3; 21.8; 23.8; and 26.1.
7. 7. The crystalline Form A of claims 3 or 4 comprising an x-ray powder diffraction pattern with characteristic peaks expressed In degrees two-theta at about 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
8. 8. The crystalline Form A of any of claims 3-7, wherein the substantially pure crystalline Form A is anhydrous.
9. 9. The crystalline Form A of any of claims 3-7, wherein the substantially pure crystalline Form A is substantially free of ethanol within its crystal system.
10. 10. The crystalline Form A of claim 3 having an onset melting temperature of between about 262°C to about 264°C,
11. 11. The crystalline Form A of claim 10 having an onset melting temperature of about 264.1°C.
12. 12. The crystalline Form A of any of claims 3-12 comprising a FT-IR spectrum with any one of the following frequency bands:
13. 13. A process for preparing crystalline Form A of 3-(imidazo[1 T2-b]pyridazin-3-ylethynyl)-4-methyt-N-{4-[(4-methylpiperazin-1-yl)methyl]'-3~{trifluoromethyl)pfienyl}benzamide mono hydrochloride comprising contacting 3-(imidazo[1,2-blpyridazin-3-ylethynyl)-4-methyl-N~{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide with hydrochloric acid.
14. 14. Crystalline Form B of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[{4-methylpiperazin-1-yl)methyl]-3-(tr!fluoromethyl)phenyl}benzamide mono hydrochloride.
15. 15. The crystalline Form B of claim 14, wherein the crystalline Form B of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantialiypure.
16. 16. The crystalline Form B of claims 14 or 15 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 15.
17. 17. The crystalline Form B of claims 14 or 15 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about: 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.
18. 18. Crystalline Form C of 3-{imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyj)phenyl}benzamide mono hydrochloride.
19. 19. The crystalline Form C of claim 18, wherein the crystalline Form C of 3-(imidazo[1,2-b]pyndazin-3-ylethynyl)-4-methyl-N-{4-[{4-melhylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
20. 20. The crystalline Form C of claims 18 or 19 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 17.
21. 21. The crystalline Form C of claims 18 or 19 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about 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.
22. 22. Crystalline Form D of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4~methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride.
23. 23. The crystalline Form D of claim 22, wherein the crystalline Form D of 3-(imidazo[1,2-b]pyridazin-3-ylethyny!)-4-methyl-N-{4-[(4-methylpiperazin-1-yS)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
24. 24. The crystalline Form D of claims 22 or 23 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 22.
25. 25. The crystalline Form D of claims 22 or 23 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about B.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.
26. 26. Crystalline Form E of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1 -yl)methyl]-3-{trifluoromethyl)phenyl}benzamide mono hydrochloride.
27. 27. The crystalline Form E of claim 26, wherein the crystalline Form E of 3-{imidazo[1,2-b]pyridazin-3-ylethynyi)-4-methyl-N-{4-[(4-methylpiperazin-1-yi)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
28. 28. Crystalline Form F of 3-(imidazo(1,2-b]pyridazin-3-ylethynyl)-4-rnethyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)pheny!}benzamide mono hydrochloride. 29 The crystalline Form F of claim 28, wherein the crystalline Form F of 3-{imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-mcthylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure. 30 The crystalline Form F of claims 28 or 29 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 27.
29. 31. The crystalline Form F of claims 28 or 29 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about 6.8; 9.8; 12.4; 16,2; 17.9; 19.0; 24.0; and 25.1.
30. 32. Crystalline Form G of 3-(imidazo[1 ,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methy!piperazin-1-yl)methyl]-3-(trifluoromethyl)phenyi}benzamide mono hydrochloride.
31. 33. The crystalline Form G of claim 32, wherein the crystalline Form G of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-I(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
32. 34. Crystalline Form H of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-{trifluoromethyl)phenyl}benzamide mono hydrochloride.
33. 35. The crystalline Form H of claim 34, wherein the crystalline Form H of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyi]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
34. 36. The crystalline Form H of claim 34 or 35 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 34.
35. 37. The crystalline Form H of claims 34 or 35 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta at about 5.9; 8.1; 9.5; 10,7; 13.4; 16.0; 17.0; 22.0; 22.8; 24.7; and 28.3.
36. 38. Crystalline Form I of 3-(im!dazo[1,2-b]pyridazin-3-ylethynyl)-4-methy!-N-{4-[(4-meihylpiperazin-1-yl)methyl]-3-(trifiuoromethyi)phenyl}benzamide mono hydrochloride,
37. 39. The crystalline Form I of claim 38, wherein the crystalline Form I of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(lrifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure,
38. 40. Crystalline Form J of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(irifluoromethyl)phenyl}benzamide mono hydrochloride.
39. 41. The crystalline Form J of claim 40, wherein the crystalline Form J of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-melhyl-N-{4-[(4-methylpiperazin-1-yl}nnethyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
40. 42. The crystalline Form J of claims 40 or 41 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 43.
41. 43. Crystalline Form K of 3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-{4-[{4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride.
42. 44. The crystalline Form K of claim 43, wherein the crystalline Form J of 3-(imidazo[1,2-b]pyridazin-3-ylethynyi)-4-methyl-N-{4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl}benzamide mono hydrochloride is substantially pure.
43. 45. The crystalline Form K of claims 43 or 44 comprising an x-ray powder diffraction pattern with characteristic peaks expressed in degrees two-theta as shown in Figure 44.