Crystalline form of JAK2 inhibitor

Abstract

To provide compounds useful as inhibitors of protein kinases, compositions thereof, and methods for treating JAK2-mediated disorders.SOLUTION: Provided is a crystalline form characterized by N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide, and a salt or hydrate thereof, as well as one or more peaks in its X-ray powder diffraction pattern.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to French application FR1902018, filed February 12, 2019, which is incorporated herein by reference in its entirety.

[0002] The present invention provides compounds and compositions thereof that are useful as inhibitors of protein kinases. [Background technology]

[0003] In recent years, the search for new therapeutic agents has been greatly aided by a better understanding of the structure of enzymes and other biomolecules associated with diseases. One important class of enzymes that has been the subject of extensive study is the protein kinases.

[0004] Protein kinases constitute a large family of structurally related enzymes involved in the control of various signaling processes in cells. Protein kinases are thought to have evolved from a common ancestral gene because their structure and catalytic function are conserved. Almost all kinases contain a similar 250-300 amino acid catalytic domain. Kinases can be classified into families by the substrates they phosphorylate (e.g., protein-tyrosine, protein-serine / threonine, lipids, etc.).

[0005] In general, protein kinases mediate intracellular signaling by effecting phosphoryl transfer from a nucleoside triphosphate to a protein acceptor that is involved in a signaling pathway. These phosphorylation events act as molecular on / off switches that can modulate or regulate the biological function of the target protein. These phosphorylation events are ultimately triggered in response to a variety of extracellular and other stimuli. Examples of such stimuli include environmental and chemical stress signals (e.g., osmotic shock, heat shock, ultraviolet radiation, bacterial endotoxin, and H2O2), cytokines (e.g., interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF-α)), and growth factors (e.g., granulocyte-macrophage colony-stimulating factor (GM-CSF) and fibroblast growth factor (FGF)). Extracellular stimuli can affect one or more cellular responses related to cell growth, migration, differentiation, secretion of hormones, activation of transcription factors, muscle contraction, glucose metabolism, control of protein synthesis, and regulation of the cell cycle.

[0006] Many diseases are related to abnormal cellular responses triggered by protein kinase-mediated events as described above.These diseases include, but are not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease, and hormone-related diseases.Therefore, there remains a need to find protein kinase inhibitors that are useful as therapeutic agents. Summary of the Invention

[0007] In some embodiments, the disclosure provides one or more crystalline forms of Compound 1. [ka]

[0008] In some embodiments, Compound 1 is useful for treating a myeloproliferative disorder. In some embodiments, the myeloproliferative disorder is selected from myelofibrosis, polycythemia vera, and essential thrombocythemia. In some embodiments, the myelofibrosis is selected from primary myelofibrosis or secondary myelofibrosis. In some embodiments, the secondary myelofibrosis is selected from post-polycythemia vera myelofibrosis and post-essential thrombocythemia myelofibrosis.

[0009] In some embodiments, the disclosure provides a method of inhibiting activity of JAK2 kinase or a mutant thereof in a biological sample, comprising contacting the biological sample with Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form), or a composition thereof.

[0010] In another embodiment, the disclosure provides a method of inhibiting activity of JAK2 kinase or a mutant thereof in a patient comprising administering to the patient Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form), or a composition thereof. In other embodiments, the disclosure provides a method of treating a JAK2-mediated disease or disorder in a patient in need thereof, the method comprising administering to the patient Compound 1, or a composition thereof. [Brief description of the drawings]

[0011] [Figure 1] 1 shows the FT-Raman spectrum of Form A of Compound 1. [Diagram 2] 1 shows the X-ray powder diffraction (XRPD) pattern of Form A of Compound 1. [Figure 3A] 1 shows a thermogravimetric analysis (TGA) pattern of Form A of Compound 1. [Figure 3B] 1 shows a differential scanning calorimetry (DSC) pattern of Form A of Compound 1. [Figure 4] 1 shows the FT-Raman spectrum of Form B of Compound 1. [Diagram 5] 1 shows the XRPD pattern of Form B of Compound 1. [Figure 6A] 1 shows a TGA pattern of Compound 1, Form B. [Figure 6B] 1 shows the DSC pattern of Form B of Compound 1. [Figure 7] 1 shows the XRPD pattern of Form C of Compound 1. [Figure 8] 1 shows the XRPD pattern of Form D of Compound 1. [Figure 9] 1 shows the XRPD pattern of Form E of Compound 1. [Figure 10] 1 shows the XRPD pattern of Form F of Compound 1. [Figure 11] 1 shows the TGA pattern of Compound 1, Form F. [Figure 12] 1 shows the dynamic vapor sorption (DVS) isotherm of Form F of Compound 1. [Figure 13] 1 shows the XRPD pattern of Form G of Compound 1. [Figure 14] 1 shows the TGA pattern of Compound 1, Form G. [Figure 15] 1 shows the DVS isotherm of Form G of Compound 1. [Figure 16] 1 shows the XRPD pattern of Form H of Compound 1. [Figure 17] 1 shows the TGA pattern of Form H of Compound 1. [Figure 18] 1 shows the DVS isotherm of Form H of Compound 1. [Figure 19] 1 shows the XRPD pattern of Form I of Compound 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] General Description of Certain Aspects of the Invention U.S. Patent 7,528,143, issued May 5, 2009 ("the '143 patent"), which is incorporated herein by reference in its entirety, describes certain 2,4-disubstituted pyrimidine compounds useful for the treatment of myeloproliferative disorders, including polycythemia vera, essential thrombocythemia, and myelofibrosis (e.g., primary myelofibrosis and secondary myelofibrosis, such as post-polycythemia vera myelofibrosis and post-essential thrombocythemia myelofibrosis). Such compounds include N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide. [ka]

[0013] N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide is designated compound number LVII, and its synthesis is detailed in Example 90 of the '143 patent.

[0014] N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide is active in a variety of assays and therapeutic models and exhibits inhibition of Janus kinase 2 (JAK2). Thus, N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide, and its salts, hydrates or solvates, are useful for treating one or more disorders associated with the activity of JAK2.

[0015] In some embodiments, the disclosure provides one or more crystalline forms of Compound 1. [ka]

[0016] It is understood that the crystalline form of Compound 1 can exist in a pure or non-solvated form, a hydrated form, and / or a solvated form. In some embodiments, the crystalline form of Compound 1 is a pure or non-solvated crystalline form, and thus does not have water or solvent incorporated into the crystal structure. In some embodiments, the crystalline form of Compound 1 is a hydrated or solvated form. In some embodiments, the crystalline form of Compound 1 is a hydrate / solvate form (also referred to herein as a "heterosolvate").

[0017] Thus, in some embodiments, the disclosure provides one or more crystalline anhydrous forms of Compound 1. [ka]

[0018] In some embodiments, the disclosure provides one or more solid hydrate forms of Compound 1. [ka]

[0019] In some embodiments, the disclosure provides one or more crystalline solvate forms of Compound 1. [ka]

[0020] In some embodiments, the present disclosure provides a substantially impurity-free sample comprising a crystalline form of Compound 1. As used herein, the term "substantially impurity-free" means that the sample does not contain a significant amount of foreign matter. In some embodiments, the sample comprising a crystalline form of Compound 1 is substantially free of amorphous Compound 1. In certain embodiments, the sample comprises at least about 90% by weight of the crystalline form of Compound 1. In certain embodiments, the sample comprises at least about 91% by weight, at least about 92% by weight, at least about 93% by weight, at least about 94% by weight of the crystalline form of Compound 1. In certain embodiments, the sample comprises at least about 95% by weight of the crystalline form of Compound 1. In yet other embodiments, the sample comprises at least about 99% by weight of the crystalline form of Compound 1.

[0021] According to some embodiments, the sample comprises at least about 95, 97, 97.5, 98.0, 98.5, 99, 99.5, 99.8 weight percent (wt%) of the crystalline form of Compound 1, where the percentage is based on the total weight of the sample. According to some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 5.0% or less of total organic impurities. In some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 3.0% or less of total organic impurities. In some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 1.5% or less of total organic impurities. In some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 1.0% or less of total organic impurities. In some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 0.6% or less of total organic impurities. In some embodiments, the sample comprising the crystalline form of Compound 1 comprises about 0.5% or less of total organic impurities. In some embodiments, the percent of total organic impurities is measured by HPLC.

[0022] Compound 1 has been found to be capable of existing in at least nine different crystalline forms, or polymorphs.

[0023] In some embodiments, the disclosure provides a crystalline hydrate form of Compound 1. In some such embodiments, the crystalline hydrate form of Compound 1 is a monohydrate. In some embodiments, the crystalline monohydrate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 9.6, 10.0, 12.4, 12.7, and 17.0±0.2 degrees 2θ. In some such embodiments, the crystalline monohydrate form of Compound 1 is Form A.

[0024] In some embodiments, Form A is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 1]

[0025] In some embodiments, Form A is characterized by the FT-Raman spectrum shown in FIG.

[0026] In some embodiments, Form A is characterized by the XRPD pattern shown in FIG.

[0027] In some embodiments, Form A is characterized by the TGA pattern shown in Figure 3 A. In some embodiments, Form A is characterized by the DSC pattern shown in Figure 3B.

[0028] In some embodiments, the disclosure provides a crystalline trihydrate form of Compound 1. In some such embodiments, the crystalline trihydrate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.4, 6.2, 11.6, 13.9, 16.4, and 16.7±0.2 degrees 2θ. In some such embodiments, the crystalline trihydrate form of Compound 1 is Form B.

[0029] In some embodiments, Form B is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 2]

[0030] In some embodiments, Form B is characterized by the FT-Raman spectrum shown in FIG.

[0031] In some embodiments, form B is characterized by the XRPD pattern shown in FIG.

[0032] In some embodiments, Form B is characterized by the TGA pattern shown in Figure 6 A. In some embodiments, Form B is characterized by the DSC pattern shown in Figure 6B.

[0033] In some embodiments, the disclosure provides a crystalline anhydrous form of Compound 1. In some such embodiments, the crystalline anhydrous form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 6.2, 8.6, 9.7, 13.6, and 17.3±0.2 degrees 2θ. In some such embodiments, the crystalline anhydrous form of Compound 1 is Form C.

[0034] In some embodiments, Form C is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 3]

[0035] In some embodiments, the disclosure provides a method for preparing a crystalline anhydrous form of Compound 1, comprising heating Form A at about 40° C. to about 80° C. under an inert atmosphere. Thus, in some embodiments, the disclosure provides a method for preparing Form C, the method comprising: (a) providing Form A; and (b) heating Form A to about 40° C. to about 80° C. under an inert atmosphere; Includes.

[0036] In some embodiments, Form C is characterized by the XRPD pattern shown in FIG.

[0037] In some embodiments, the disclosure provides a crystalline anhydrous form of Compound 1 characterized by one or more peaks in its X-ray powder diffraction pattern selected from 12.8, 13.6, 14.9, 16.1, and 17.2±0.2 degrees 2θ. In some such embodiments, the crystalline anhydrous form of Compound 1 is Form D.

[0038] In some embodiments, Form D is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 4]

[0039] In some embodiments, the disclosure provides a method for preparing a crystalline anhydrous form of Compound 1, comprising heating Form B at about 25° C. to about 70° C. under an inert atmosphere. Thus, in some embodiments, the disclosure provides a method for preparing Form D, the method comprising: (c) providing Form B; and (d) heating Form B to about 25° C. to about 70° C. under an inert atmosphere. Includes.

[0040] In some embodiments, form D is characterized by the XRPD pattern shown in FIG.

[0041] In some embodiments, the disclosure provides a crystalline solvate form of Compound 1. In some such embodiments, the crystalline solvate form of Compound 1 is a monosolvate. In some embodiments, the crystalline monosolvate form of Compound 1 is a monoisopropanol solvate. In some such embodiments, the crystalline monoisopropanol solvate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 7.2, 10.4, 10.8, 13.2, and 17.5±0.2 degrees 2θ. In some such embodiments, the crystalline monoisopropanol solvate form of Compound 1 is Form E.

[0042] In some embodiments, Form E is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 5]

[0043] In some embodiments, form E is characterized by the XRPD pattern shown in FIG.

[0044] In some embodiments, the crystalline solvate form of Compound 1 is a tetrasolvate. In some embodiments, the crystalline tetrasolvate form of Compound 1 is a tetraisopropanol solvate. In some embodiments, the crystalline tetraisopropanol solvate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.3, 6.4, 8.2, 10.5, 15.3, and 15.7±0.2 degrees 2θ. In some such embodiments, the crystalline tetraisopropanol solvate form of Compound 1 is Form F.

[0045] In some embodiments, Form F is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 6]

[0046] In some embodiments, form F is characterized by the XRPD pattern shown in FIG.

[0047] In some embodiments, form F is characterized by the TGA pattern shown in FIG.

[0048] In some embodiments, form F is characterized by the DVS isotherm shown in FIG.

[0049] In some embodiments, the crystalline solvate form of Compound 1 is a heterosolvate. In some embodiments, the crystalline heterosolvate form of Compound 1 is a water-isopropanol heterosolvate. In some embodiments, the crystalline water-isopropanol heterosolvate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 7.3, 7.6, 10.4, 10.8, and 17.5±0.2 degrees 2θ. In some such embodiments, the crystalline water-isopropanol heterosolvate form of Compound 1 is Form G.

[0050] In some embodiments, Form G is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 7]

[0051] In some embodiments, form G is characterized by the XRPD pattern shown in FIG.

[0052] In some embodiments, Form G is characterized by the TGA pattern shown in FIG.

[0053] In some embodiments, form G is characterized by the DVS isotherm shown in FIG.

[0054] In some embodiments, the crystalline solvate form of Compound 1 is a hexafluoroisopropanol solvate. In some embodiments, the crystalline hexafluoroisopropanol solvate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 6.0, 6.9, 10.9, 11.5, 14.7, and 17.1±0.2 degrees 2θ. In some such embodiments, the crystalline hexafluoroisopropanol solvate form of Compound 1 is Form H.

[0055] In some embodiments, Form H is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 8]

[0056] In some embodiments, form H is characterized by the XRPD pattern shown in FIG.

[0057] In some embodiments, form H is characterized by the TGA pattern shown in FIG.

[0058] In some embodiments, form H is characterized by the DVS isotherm shown in FIG.

[0059] In some embodiments, the crystalline solvate form of Compound 1 is an ethanol solvate. In some embodiments, the crystalline ethanol solvate form of Compound 1 is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.3, 10.6, and 15.9±0.2 degrees 2θ. In some such embodiments, the crystalline ethanol solvate form of Compound 1 is Form I.

[0060] In some embodiments, Form I is characterized by the following peaks in its X-ray powder diffraction pattern: [Table 9]

[0061] In some embodiments, Form I is characterized by the XRPD pattern shown in FIG.

[0062] Uses, Formulation and Administration Pharmaceutically acceptable compositions According to another embodiment, the present disclosure provides a composition comprising Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) and a pharma- ceutically acceptable carrier, adjuvant, or vehicle. In certain embodiments, the amount of Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) in the composition of the present disclosure is such that it is effective to measurably inhibit JAK2 or a mutant thereof in a biological sample or a patient. In certain embodiments, the composition of the present disclosure is formulated for administration to a patient in need of such a composition. In some embodiments, the composition of the present disclosure is formulated for oral administration to a patient.

[0063] The compounds and compositions, according to the methods of the invention, are administered in any amount and using any route of administration effective to treat or reduce the severity of the disorders provided herein (i.e., JAK2-mediated diseases or disorders). The exact amount required will vary from subject to subject, depending on the subject's race, age, and general condition, the severity of the infection, the particular agent, its mode of administration, and the like. Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) is preferably formulated in unit dosage form for ease of administration and uniformity of dosage.

[0064] The compositions of the present disclosure may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, intraperitoneally, intracisternally, or via an implanted reservoir. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously.

[0065] The sterile injectable form of the composition of the present disclosure may be an aqueous or oily suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be utilized are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspending medium.

[0066] For this purpose, any non-irritating fixed oil can be used, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful for the preparation of injectables, as well as natural pharmaceutically acceptable oils such as olive oil or castor oil, especially in their polyoxyethylated forms. These oil solutions or suspensions may contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose, or similar dispersants commonly used in the preparation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers, commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for formulation purposes.

[0067] Injectable preparations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0068] In order to prolong the effect of compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form), it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The absorption rate of compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) will therefore depend on its dissolution rate, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the specific polymer used, the release rate of compound can be controlled.Other examples of biodegradable polymers include poly(orthoester) and poly(anhydride).Injectable depot formulations can also be prepared by encapsulating Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) in liposome or microemulsion that is compatible with body tissue.

[0069] In some embodiments, the pharma- ceutically acceptable compositions provided are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharma- ceutically acceptable compositions of the present disclosure are administered without food. In other embodiments, the pharma- ceutically acceptable compositions of the present disclosure are administered with food.

[0070] The pharma- ceutically acceptable composition of the present disclosure can be orally administered in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions.For tablets for oral use, commonly used carriers include lactose and cornstarch.Lubricants such as magnesium stearate are also generally added.For oral administration in capsule form, useful diluents include lactose and dry cornstarch.When aqueous suspensions are required for oral use, active ingredient is combined with emulsifying and suspending agents.If desired, certain sweeteners, flavors, or colorants may be added.

[0071] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) is mixed with at least one inert pharma- ceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) glycerol, and the like. humectants, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and / or i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0072] Solid compositions of a similar type may also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and may also be part of the composition so as to release the active ingredient(s) only in a certain part of the intestinal tract, i.e. preferentially, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric substances and waxes. Solid compositions of a similar type may also be used as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols.

[0073] Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) may be in microencapsulated form with one or more of the above excipients. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) can be mixed with at least one inert diluent, such as sucrose, lactose, or starch. According to conventional methods, such dosage forms can also contain additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms can also contain buffering agents. They may optionally contain opacifying agents or may be of a composition that they release the active ingredient(s) only in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0074] Liquid dosage forms for oral administration include, but are not limited to, pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.In addition to Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form), liquid dosage forms may contain, for example, inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0075] Alternatively, the pharma- ceutically acceptable compositions of the present disclosure may be administered in the form of suppositories for rectal administration. These may be prepared by mixing Compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, thereby melting in the rectum and releasing the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0076] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate form) with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, or a suppository wax, which is solid at ambient temperature but liquid at body temperature and thus will melt in the rectal or vaginal cavity and release the active compound therein.

[0077] The pharma- ceutically acceptable compositions of the present disclosure may be administered topically, particularly when the target of treatment includes areas or organs readily accessible by topical application, such as diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0078] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topical-transdermal patches may also be used.

[0079] For topical application, the provided pharma- ceutically acceptable composition can be formulated into a suitable ointment containing compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) suspended or dissolved in one or more carriers. Carriers for topical administration of compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax, and water. Alternatively, the provided pharma-ceutically acceptable composition can be formulated into a suitable lotion or cream containing compound 1 (e.g., crystalline anhydrous form, crystalline hydrate form, crystalline solvate form, or crystalline heterosolvate form) suspended or dissolved in one or more pharma-ceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0080] For ophthalmic use, the pharma- ceutically acceptable compositions provided may be formulated as a micronized suspension in isotonic, pH-adjusted, sterile saline or, preferably, as an isotonic, pH-adjusted, sterile saline solution, with or without a preservative, such as benzylalkonium chloride. Alternatively, for ophthalmic use, the pharma-ceutically acceptable compositions may be formulated in an ointment, such as petrolatum.

[0081] Pharmaceutically acceptable compositions of the invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0082] Dosage forms for topical or transdermal administration of Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate forms) include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate forms) is mixed under sterile conditions with a pharma- ceutically acceptable carrier and, if necessary, any necessary preservatives or buffers. Ophthalmic formulations, ear drops, and eye drops are also contemplated to be within the scope of the present invention. In addition, the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing a controlled delivery of Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate forms) to the body. Such dosage forms can be made by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate forms) across the skin. The rate can be controlled by either using a rate controlling membrane or by dispersing Compound 1 (e.g., crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate forms) in a polymer matrix or gel.

[0083] In some embodiments, the compositions described herein contain a molar equivalent amount of Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) to the free base N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide. For example, a 100 mg formulation of N-tert-butyl-3-[(5-methyl-2-{[4-(2-pyrrolidin-1-ylethoxy)phenyl]amino}pyrimidin-4-yl)amino]benzenesulfonamide (i.e., the unsolvated free base parent of Compound 1, MW=524.26) contains 117.30 mg of Form A (MW=614.22).

[0084] In some embodiments, the present disclosure provides compositions comprising Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) and one or more pharma- ceutically acceptable excipients. In some embodiments, the one or more pharma-ceutically acceptable excipients are selected from binders and lubricants.

[0085] In one embodiment, the binder is microcrystalline cellulose. In some such embodiments, the microcrystalline cellulose is silicified microcrystalline cellulose.

[0086] In some embodiments, the binder is sodium stearyl fumarate.

[0087] In some embodiments, the composition comprises: [Table 10]

[0088] In certain embodiments, the composition comprises: [Table 11]

[0089] Uses of the Compounds and Pharmaceutically Acceptable Compositions The compounds and compositions described herein are generally useful for inhibiting the kinase activity of one or more enzymes. Examples of kinases that are inhibited by the compounds and compositions described herein and for which the methods described herein are useful include JAK2 or mutants thereof.

[0090] The activity of Compound 1 (e.g., in a crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate form) utilized as an inhibitor of JAK2 kinase or a mutant thereof can be assayed in vitro, in vivo, or in a cell line. In vitro assays include assays that determine inhibition of either the phosphorylation activity and / or subsequent functional consequences, or ATPase activity, of activated JAK2 kinase or a mutant thereof.

[0091] According to one embodiment, the present invention relates to a method of inhibiting protein kinase activity in a biological sample, the method comprising the step of contacting said biological sample with Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form), or a composition thereof.

[0092] According to another embodiment, the present invention provides a method of inhibiting activity of JAK2 kinase or a mutant thereof in a biological sample, the method comprising contacting said biological sample with Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form), or a composition thereof.

[0093] According to another embodiment, the present invention relates to a method of inhibiting activity of JAK2 kinase or a mutant thereof in a patient, comprising administering Compound 1 (e.g., in a crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate form), or a composition thereof, to said patient. In other embodiments, the disclosure provides a method for treating a JAK2-mediated disease or disorder in a patient in need thereof, comprising administering Compound 1 (e.g., in a crystalline anhydrous, crystalline hydrate, crystalline solvate, or crystalline heterosolvate form), or a pharma- ceutically acceptable composition thereof, to said patient. Such disorders are described in detail herein.

[0094] The crystalline forms described herein are useful for treating a variety of disorders, including, but not limited to, myeloproliferative disorders, proliferative diabetic retinopathy, and other angiogenesis-related disorders, including solid tumors and other types of cancer, eye diseases, inflammation, psoriasis, and viral infections. Types of cancer that can be treated include, but are not limited to, gastrointestinal / gastrointestinal tract cancer, colon cancer, liver cancer, skin cancer, breast cancer, ovarian cancer, prostate cancer, lymphoma, leukemia (acute myeloid leukemia and chronic myeloid leukemia), kidney cancer, lung cancer, muscle cancer, bone cancer, bladder cancer, or brain cancer.

[0095] Some examples of diseases and disorders that can be treated include ocular angiogenesis, infantile hemangioma; organ hypoxia, vascular hyperplasia, organ transplant rejection, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type 1 diabetes and diabetic complications, inflammatory diseases, acute pancreatitis, chronic pancreatitis, asthma, allergies, adult respiratory distress syndrome, cardiovascular disease, liver disease, other blood diseases, asthma, rhinitis, atopic dermatitis, dermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, metastatic melanoma, Kaposi's sarcoma, multiple myeloma, cytokine-related conditions, as well as glomerulonephritis, sclerosing dermatitis, chronic thyroiditis, Graves' disease, autoimmune gastritis, autoimmune hemolytic anemia, autoimmune rheumatoid arthritis, and autoimmune thyroiditis. Neurodegenerative diseases including neutropenia, thrombocytopenia, atopy (e.g., allergic asthma, atopic dermatitis, or allergic rhinitis), chronic active hepatitis, gravitational myasthenia, multiple sclerosis, inflammatory bowel disease, other autoimmune diseases including graft versus host disease, motor neuron disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia, or neurodegenerative diseases caused by traumatic injury, strike, glutamate neurotoxicity or hypoxia; also included are stroke, myocardial ischemia, renal ischemia, heart attack, cardiac hypertrophy, atherosclerosis and arteriosclerosis, organ hypoxia, and ischemic / reperfusion injury in platelet aggregation.

[0096] Some additional diseases and disorders that can be treated also include cell-mediated hypersensitivity (allergic contact dermatitis, hypersensitivity pneumonitis), rheumatic diseases (e.g., systemic lupus erythematosus (SLE), juvenile arthritis, Sjogren's syndrome, scleroderma, ankylosing spondylitis, psoriatic arthritis), viral diseases (Epstein-Barr virus, Hepatitis B, Hepatitis C, HIV, HTLVI, Varicella-Zoster virus, Human papilloma virus), food allergies, skin inflammation, and immunosuppression induced by solid tumors.

[0097] In some embodiments, Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) is useful for treating a myeloproliferative disorder. In some embodiments, the myeloproliferative disorder is selected from primary myelofibrosis, polycythemia vera, and essential thrombocythemia. In some embodiments, the myeloproliferative disorder is selected from primary myelofibrosis and secondary myelofibrosis. In some such embodiments, the myeloproliferative disorder is secondary myelofibrosis. In some such embodiments, the secondary myelofibrosis is selected from post-polycythemia vera myelofibrosis and post-essential thrombocythemia myelofibrosis.

[0098] In some embodiments, the methods provided include administering Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) to a patient previously treated with a JAK2 inhibitor. In some such embodiments, the methods provided include administering Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) to a patient previously treated with ruxolitinib (JAKAFI®).

[0099] In some embodiments, methods provided include administering Compound 1 (e.g., a crystalline anhydrous form, a crystalline hydrate form, a crystalline solvate form, or a crystalline heterosolvate form) to a patient suffering from or diagnosed with a myeloproliferative disorder unresponsive to ruxolitinib. In some embodiments, the patient suffers from or has been diagnosed with a myeloproliferative disorder that is refractory or resistant to ruxolitinib.

[0100] In some embodiments, the patient relapses during or after ruxolitinib treatment.

[0101] In some embodiments, the patient is intolerant to ruxolitinib. In some embodiments, the patient's intolerance to ruxolitinib is evidenced by hematological toxicity (e.g., anemia, thrombocytopenia, etc.) or non-hematological toxicity.

[0102] In some embodiments, the patient has had an inadequate response to or is intolerant to hydroxyurea.

[0103] In some embodiments, the patient exhibits or experiences, or has exhibited or experienced, one or more of the following during treatment with ruxolitinib: lack of response, disease progression, or loss of response at any time during ruxolitinib treatment. In some embodiments, disease progression is evidenced by an increase in spleen size during ruxolitinib treatment.

[0104] In some embodiments, the patient previously treated with ruxolitinib has a somatic mutation or clonal marker associated with or indicative of a myeloproliferative disorder. In some embodiments, the somatic mutation is selected from a JAK2 mutation, a CALR mutation, or an MPL mutation. In some embodiments, the JAK2 mutation is V617F. In some embodiments, the CALR mutation is a mutation in exon 9. In some embodiments, the MPL mutation is selected from W515K and W515L.

[0105] In some embodiments, the disclosure provides methods of treating a relapsed or refractory myeloproliferative disorder, wherein the myeloproliferative disorder is relapsed or refractory to ruxolitinib.

[0106] In some embodiments, the myeloproliferative disorder is selected from intermediate risk myelofibrosis and high risk myelofibrosis.

[0107] In some embodiments, the intermediate risk myelofibrosis is selected from primary myelofibrosis, post-polycythemia vera (post-PV) myelofibrosis, and post-essential thrombocythemia (post-ET) myelofibrosis. In some embodiments, the myelofibrosis is intermediate risk 1 (also referred to as intermediate 1 risk). In some embodiments, the myelofibrosis is intermediate risk 2 (also referred to as intermediate 2 risk).

[0108] In some embodiments, the high-risk myelofibrosis is selected from primary myelofibrosis, post-polycythemia vera (post-PV) myelofibrosis, and post-essential thrombocythemia (post-ET) myelofibrosis.

[0109] In some embodiments, the disclosure provides an article of manufacture comprising packaging material and a pharmaceutical composition contained within the packaging material, hi some embodiments, the packaging material comprises a label indicating that the pharmaceutical composition can be used to treat one or more of the disorders identified above.

[0110] Example 1: Solubility evaluation Solubility was evaluated in a diverse set of solvents to facilitate the selection of solvent systems and corresponding dosing strategies for subsequent crystal form screening experiments. The solubility of Form A was visually estimated in 12 solvents by injecting a small amount of solvent into a fixed amount of API (10.0 mg) until the dissolution point or a maximum volume of 1.8 mL was reached at room temperature and, where applicable, 40° C. As shown in Table 1, Form A exhibits high solubility (>100 mg / mL) in DMSO, MeOH, and water, but low solubility (<5 mg / mL) in all other solvents evaluated. Table 1: Estimated solubility of Form A in 12 solvents at room temperature and 40°C. [Table 12]

[0111] Example 2: Crystal Morphology Screen FT-Raman spectroscopy: Raman spectra were collected using a Nicolet NXR9650 or NXR 960 spectrometer (Thermo Electron) equipped with a 1064 nm Nd:YVO4 excitation laser, InGaAs and liquid N2-cooled Ge detectors, and a MicroStage. All spectra were measured at 4 cm- through a glass cover using the Happ-Genzel apodization function and two levels of zero-filling. 1 The image was acquired in 64 scans at 100x the resolution.

[0112] X-ray powder diffraction (PXRD): PXRD diffractograms were obtained on a PANalytical X' Pert Pro diffractometer using Ni filtered CuKa (45 kV / 40 mA) radiation and a step size of 0.02° 2θ with an X'celerator™ RTMS (real time multistrip) detector. Incident beam configuration: fixed divergence slit (0.25°), 0.04 rad Soller slit, anti-scatter slit (0.25°), and 10 mm beam mask. Diffracted beam configuration: fixed divergence slit (0.25°) and 0.04 rad Soller lit.

[0113] Differential Scanning Calorimetry (DSC): DSC was performed using a TA Instruments Q100 differential scanning calorimeter equipped with an autosampler and a refrigerated cooling system with a N2 purge of <40 mL / min. Unless otherwise stated, DSC thermograms were obtained in crimped Al pans at 15 °C / min.

[0114] Thermogravimetric analysis (TGA): Unless otherwise stated, TGA thermograms were obtained at 15 °C / min in Al pans with a N2 purge of less than 40 mL / min using a TA Instruments Q500 thermogravimetric analyzer.

[0115] Thermogravimetric analysis with IR off-gas detection (TGA-IR): TGA-IR was performed using a TA Instruments Q5000 thermogravimetric analyzer connected to a Nicolet 6700 FT-IR spectrometer (Thermo Electron) equipped with an external TGA-IR module with a gas flow cell and a DTGS detector. Unless otherwise stated, TGA was performed in Pt or Al pans with a N2 flow rate of 60 mL / min and a heating rate of 15 °C / min. IR spectra were recorded at 4 cm- 1 The images were collected at a resolution of 100x and 32 scans.

[0116] Modulated Differential Scanning Calorimetry (mDSC): mDSC was performed using a TA Instruments Q200 differential scanning calorimeter equipped with an autosampler and a refrigerated cooling system under a 40 mL / min N2 purge. mDSC thermograms were acquired using a modulation of ±0.32°C every 60 seconds, followed by an isothermal hold for 5.00 minutes, followed by a heating ramp to 200°C at 2.00°C / min. Sample preparation was performed in crimped Al pans.

[0117] Freeze drying: Freeze drying was performed on a Virtis Lyo-Centre Benchtop 3.5DBTZL, serial number: 41712. The unit was operated at a pressure below 10 mtorr and a condenser temperature below -100°C.

[0118] Ion Chromatography (IC): Ion chromatography was performed on a Dionex ICS-3000. Column: Dionex IonPac AS12A 4x200mm. Detection: ASRS 300 with suppressed conductivity, 22mA suppressor current. Eluent (2.7mM Na2CO3 / 0.3mM NaHCO3) at 1.5mL / min.

[0119] Solvent Selection: The crystal morphology screen involved 48 solvent systems. Solvents were utilized as pure and binary mixtures to provide a diverse set of polarity, dielectric constant, dipole moment, and hydrogen bond donor / acceptor properties. Hydrous solvents with a range of water activities were also included.

[0120] Crystallization Mode: The following crystallization modes were employed in the crystal morphology screening study using amorphous input material: (1) Stir the suspension while cycling the temperature between 45°C and 5°C for 3 days (TC, n=48) (2) The clarified solution was cooled from 45°C to 5°C and then held for 4 days (RC, n=48) (3) Slowly evaporate the solvent from the solution over 7–10 days at room temperature (EV, n = 48; Form A input).

[0121] Analysis of Screened Products: FT-Raman spectroscopy was selected as the primary method for analysis and grouping of samples. Representative samples from the groupings were analyzed by PXRD to verify their uniqueness. Where possible / practical, representative samples of unique morphology were further characterized by polarized light microscopy, DSC, and / or TGA-IR.

[0122] Screen Results: Form A was observed as the primary output of the screening experiments. Form B, a hydrated form, was observed in 15 different slurry and evaporation experiments. As shown in Table 2, the crystal form screen for Compound 1 produced the following forms: Table 2 - Preliminary crystal morphology screen [Table 13] [Table 14]

[0123] Characterization of Form A: Form A is a white powder and was determined to be crystalline by Raman (Figure 1) and PXRD analysis (Figure 2). DSC shows a broad, shallow endotherm from 25 to 150 °C followed by a decomposition endotherm at 216.4 °C (Figure 3B). TGA-IR analysis showed the release of 2.9% water from 25 to 150 °C (1 equiv, monohydrate) corresponding to the broad DSC endotherm (Figure 3A).

[0124] Characterization of Form B: Form B is a hydrate form observed during screening. Raman (Figure 4) and PXRD (Figure 5) analysis indicate that Form B is crystalline. DSC shows a broad endotherm between 25 and 110 °C (Figure 6B) associated with a 9.4% weight loss (3.4 equiv.) of water by TGA-IR (Figure 6A). DSC analysis also shows a small, low energy endotherm at 147.7 °C. Form B was confirmed by IC to be the di-HCl salt.

[0125] Example 3. Comparative stability study of Forms A and B Relative stability studies were performed on monohydrate form A and hydrate form B at 25° C. to determine stable hydrates at various water activity levels.

[0126] A saturated suspension of Form A was prepared by stirring an excess of Form A in the specified solvent system. The suspension was stirred overnight at 25° C. A clarification filtration was performed and the filtrate was added to a 2 mL vial containing approximately 10 mg of Form A and approximately 10 mg of Form B. The resulting suspension was stirred at 25° C. for 7 days, isolated, dried under vacuum for 45 minutes, and analyzed by FT-Raman.

[0127] Form A was obtained after aging studies. The summary results of the studies are shown in Table 3 and show that Form A is a stable hydrate at 25° C. over the entire water activity range. Table 3 - Results of competitive ripening study [Table 15]

[0128] Example 4. Additional Crystal Morphology Screen measurement High-resolution X-ray powder diffraction (high-resolution XRPD): High-resolution diagrams are recorded in ambient conditions on a Panalytical X'Pert Pro MPD powder diffractometer using a Bragg-Brentano (vertical θ-2θ configuration) parafocusing geometry in combination with an X'Celerator detector. A sealed copper anode X-ray tube is used, operating at 45 kV and 40 mA levels. An incident beam monochromator (Johansson type: symmetrically cut curved germanium (111) crystal) produces pure CuΚα1 radiation (λ=1.54060 Å). A thin layer of the product is deposited on a single-crystal silicon wafer, cut according to the Si(510) crystallographic orientation, which prevents the Bragg reflection by systematic annihilation. To bring more crystallites into the diffraction position and thus reduce the influence of particle statistics on the measurements, a sample spinner is used. The rotation speed of the spinner is set to one revolution per second. The angular range spans from 2 to 50° in 2θ with a step size of 0.017° in 2θ. Variable count times from 500 to 5000 s per step were used.

[0129] X-ray powder diffraction (XRPD): XRPD analysis is performed on a Siemens-Bruker D5000 Matic powder diffractometer using a Bragg-Brentano (vertical θ-2θ configuration) parafocusing geometry. A sample feeder allows the automation of the work. If a sufficient amount of product is available, the powder is top-loaded into a concave stainless steel sample holder. Otherwise, a thin layer of the product is deposited on a single crystal silicon wafer, cut according to the Si(510) crystallographic orientation, which prevents the Bragg reflection by systematic extinction. A sealed cobalt anode X-ray tube operating at 45 kV and 40 mA levels is used. Two lines are usually emitted: CoKα1 (λ=1.7890 Å) and CoKα2 (λ=1.7929 Å). An iron β-filter placed between the detector and the sample does not completely remove the CoKβ (λ=1.6208 Å) radiation, which contributes about 1% of the diffracted beam at the detector (manufacturer's data). The primary beam passes through a parallel plate collimator (0.2 mm Soller slits) and then a divergence slit (0.2 mm). A Brown 50M multicanal linear detector completes the setup. This detector has a detection window 8° wide in angle 2θ. Diagrams should be recorded with a scan from 2 to 50.0° in angle 2θ, a count time of 20 s per degree in 2θ, and ambient environmental conditions such as pressure, temperature and relative humidity.

[0130] Temperature and relative humidity X-ray powder diffraction: The tests are performed using a Siemens-Bruker D5000 diffractometer equipped with a Bragg-Brentano parafocusing (θ-θ) geometry and an Anton-Paar TTK450 temperature chamber. In certain tests, dry nitrogen or RH streams are used. The powder is placed in a concave stainless steel sample holder. A sealed cobalt anode X-ray tube operated at 45 kV and 40 mA levels is used. Typically, two lines are emitted: CoKα1 (λ=1.7890 Å) and CoKα2 (λ=1.7929 Å). An iron β-filter placed between the detector and the sample does not completely eliminate the CoKβ (λ=1.6208 Å) radiation, which contributes about 1% of the diffracted beam at the detector (manufacturer's data). The beam is collimated using Soller slits to improve parallelism. A variable divergence slit keeps the illuminated area of ​​the sample constant. A 1 mm collimator limits diffusion between the tube and the sample. A Brown 50M multicanal linear detector completes the setup. This detector has a detection window 8° wide at angle 2θ. The temperature is increased at a rate of 0.05°C / s. Diagrams are typically recorded with a scan from 1.5 to 50.0 degrees at angle 2θ and a count time of 10-15 seconds per degree at 2θ. Once the requested temperature is reached, data are acquired in isothermal mode.

[0131] Simultaneous thermogravimetric analysis coupled to FTIR spectrometer (TGA-FTIR): The analysis is performed using a TG209C Netzsch Instrument coupled with a Tensor 27 Bruker FTIR spectrometer. This system allows simultaneous thermogravimetric analysis (TGA) and FTIR chemical identification of evolved compounds (water and solvent). The evolved gases are transported to the FTIR spectrometer via a transfer line heated to 476 K to prevent condensation of evolved products. A sample mass of 5-10 mg is deposited in an aluminum crucible. The TGA-FTIR analysis is performed under a dry nitrogen flow of 10 mL / min. Typically, the sample is heated from 298 to 520-570 K at a rate of 5 K / min. From 4000 to 700 cm- 1in the spectral region, 4cm- 1 A resolution of 1000 nm and 20 scans / spectrum are used to record the FTIR spectra. For each solvent analyzed by FTIR, a specific wavenumber range related to the type of solvent must be selected.

[0132] Thermogravimetric analysis: The analysis is performed on a TA Instruments TGAQ500 or TGAQ5000 analyzer. Mass calibration is performed with certified masses of 10 and 100 mg and the instrument is temperature calibrated with Alumel and Nickel standards (Curie points of 154 °C and 354 °C, respectively). Samples are exposed to a constant nitrogen flow of 60 mL / min and the temperature range is 20-250 °C at a rate of 5 °C / min. The amount of product is between 2-5 mg. The powder is deposited in an open aluminum sample pan which is placed in a platinum pan.

[0133] Differential Scanning Calorimetry (DSC): The analysis is performed under nitrogen flow using a TA Instruments Q1000 (or Q200) analyzer. The calorimeter is temperature calibrated with indium and lead (start temperatures are 156.6 °C and 327.5 °C, respectively). Energy calibration is performed using a certified indium calibrator (enthalpy of fusion of 28.45 J / g). A mechanical compressor is used to acquire and equilibrate a temperature program from 0 to 270 °C at a rate of 5 °C / min under a constant nitrogen flow of 55 mL / min (50 mL / min, respectively). The amount of product to be analyzed is 1 to 5 mg and is placed in a crimped or aluminum sample pan.

[0134] Moisture sorption / desorption isotherms: All experiments were performed on a DVS-1 automated gravimetric vapor sorption analyzer (Surface Measurement Systems Ltd., London, UK). The DVS measures vapor uptake and loss gravimetrically using a Cahn D200 recording ultramicrobalance with a mass resolution of ±0.1 μg. Controlled relative humidity is generated by mixing varying ratios of dry and water-saturated carrier gas streams (monitored by a mass flow controller). Temperature was kept constant at ±0.1 °C by enclosing the entire system in a temperature-controlled incubator. Sample sizes of approximately 10 mg were used. Prior to exposure to water vapor, samples were dried at 0% relative humidity (RH) to remove any surface water present and to establish a dry baseline mass. Samples were then exposed to increasing relative humidity, where the relative humidity was increased from 0% to 95% RH (or 90% RH) in 5% RH increments. At each stage, the sample mass was allowed to reach equilibrium before the relative humidity was increased or decreased (considering equilibrium established when the dm / dt ratio (m = mass, NS = time) did not exceed a value of 3.3 10-4 mg / s for 30 min). If equilibrium was not reached, a change in relative humidity occurred automatically after 600 min. Two consecutive cycles were recorded. From the complete moisture sorption and desorption profiles, isotherms were calculated using DVS Advanced Analysis Suite v3.6. All experiments were carried out at 25.0 °C.

[0135] Polymorph screening was performed at a variety of solvents, supersaturations, temperatures, and water activities. Slow evaporation at room temperature, evaporation at elevated temperatures under atmospheric pressure or vacuum, dissolution at reflux temperature followed by crystallization by slow cooling to room temperature Precipitation by addition of non-solvent, solvent transfer by azeotropic distillation Slurrying in pure form or mixtures of forms in anhydrous or aqueous solvents at room temperature The condition was used.

[0136] The polymorphs identified by the crystallization studies using the XRPD results are shown in Table 4. Table 4: Results of crystallization studies [Table 16]

[0137] Example 5: Variable Temperature T-XRPD Analysis of Form A and Form B: The crystalline form identified in Example 4 was characterized as follows.

[0138] The structural behavior of Form A under heating was studied in situ by T-XRPD under nitrogen atmosphere. The temperature was increased from room temperature to 230 °C in 10 °C increments. Structural changes in the XRPD diagram due to heat are observed (Figure 7). From 40 °C to 80 °C, the appearance of Form C is recorded, corresponding to the dehydrated form of Form A. Then, between 80 °C and 90 °C, a structural change is observed. At 90 °C, a new XRPD pattern appears, corresponding to another anhydrous crystalline phase. Between 160 °C and 170 °C, a major structural change is observed.

[0139] The structural behavior of form B under heating was studied in situ by T-XRPD under nitrogen atmosphere. The temperature was increased from room temperature to 160 °C in 10 °C increments. Changes in the XRPD diagram due to thermal loss of water molecules and expansion of the crystal lattice were observed from room temperature to 70 °C, resulting in the formation of form D (Figure 8). Between 90 °C and 120 °C, no structural changes were observed. Finally, at 130 °C, the XRPD diagram is flat, resembling an amorphous material.

[0140] Example 6: Characterization of Crystalline Morphology The crystalline form identified in Example 4 was characterized as follows.

[0141] The XRPD of Form F: tetraisopropanol solvate is shown in FIG.

[0142] DVS of the tetraisopropanol solvate was performed at 25 °C after a "0%P / P0" pretreatment at 25 °C (Figure 12). When the sample was exposed to increasing partial pressures of isopropanol, very slight and continuous isopropanol adsorption was observed from 0 to 45%P / P 0(IPA) (45%P / P 0(IPA) 2.1%). 50~55%P / P 0(IPA) Between 60%P / P a significant isopropanol uptake of up to 25% is observed. 0(IPA) from 90% P / P 0(IPA) Between 90%P / P, a continuous isopropanol uptake of up to 3% is observed. 0(IPA) and 40% P / P 0(IPA) A very slight and continuing loss of isopropanol amounting to 3% is observed between 35% P / P 0(IPA) and 0% P / P 0(IPA) A very significant loss of isopropanol is observed, reaching 30%. XRPD of the tetraisopropanol solvate sample after DVS revealed a new form, which was characterized by XRPD as monoisopropanol solvate form E (Figure 9).

[0143] The TGA-IR curve of Form F is shown in Figure 11. A first weight loss of 22%, corresponding to 3 moles of isopropanol, is recorded from room temperature to about 90°C. A second weight loss of 7%, corresponding to 1 mole of isopropanol, is recorded from 90°C to about 160°C. Above 210°C, thermal decomposition is observed.

[0144] The XRPD pattern of Form G: water:isopropanol heterosolvate is shown in FIG.

[0145] The TGA-IR curve of the water:isopropanol heterosolvate is shown in Figure 14. A first weight loss of 3%, corresponding to the adsorption of water, is recorded from room temperature to about 100°C. A second weight loss of 5%, corresponding to 0.5 moles of isopropanol, is recorded from 100°C to about 160°C. Above 210°C, thermal decomposition is observed.

[0146] DVS of the water:isopropanol heterosolvate is performed at 25 °C after a "0% RH" pretreatment at 25 °C (partial dehydration, observed for pretreatment at 25 °C) (Figure 15). After 1 day of exposure at 25 °C and 0% RH, partial dehydration and desolvation of the sample is observed, with a loss of 7.7% of water and IPA. When the sample is exposed to increasing relative humidity, significant and continuous water adsorption is measured between 5 and 65% RH (7.4% at 65% RH). Between 70 and 90% RH, a slight and continuous water uptake is observed, reaching 0.7%. Between 90% RH and 10% RH, a continuous water uptake is observed, reaching 2.5%. At 10% RH, a significant water loss is measured, reaching 7.1%. At 0% RH, a difference between the first and second cycle is recorded (reaching 0.6%), which may correspond to a structural change of the crystalline phase.

[0147] The XRPD pattern of Form H: hexafluoroisopropanol solvate is shown in FIG.

[0148] The TGA curve of the hexafluoroisopropanol solvate is shown in Figure 17. A first weight loss of 14.8% is recorded from 60°C to about 100°C (likely corresponding to 0.5 moles of hexafluoroisopropanol, no TGA-IR or TGA-MS measurements were performed). A second weight loss of 8.5% is recorded from 100°C to about 160°C. Above 180°C, thermal decomposition is observed.

[0149] A DVS curve is performed at 25 °C in hexafluoroisopropanol solvate (Figure 18). After 6 h under nitrogen, a weight loss of 5% is recorded (partial desolvation is observed with pretreatment at 25 °C). During the first adsorption cycle, a significant weight loss is recorded from 0 to 75% RH (approximately 30%). This weight loss corresponds to a structural change indicating desolvation due to solvent exchange induced by water vapor. It is confirmed that form H is transformed into form B.

[0150] The XRPD pattern of Form I: Ethanol solvate is shown in Figure 19. Form I is a weatherable solvate under ambient conditions.

Claims

1. Crystalline forms of Compound 1. 【Chemistry 1】

2. 2. The crystalline form of claim 1, wherein the form is a hydrate.

3. 3. The crystalline form of claim 2, wherein the form is a monohydrate.

4. 3. The crystalline form of claim 2, wherein the monohydrate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 9.6, 10.0, 12.4, 12.7, and 17.0±0.2 degrees 2θ.

5. 3. The crystalline form of claim 2, wherein the monohydrate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 1】

6. 3. The crystalline form of claim 2, wherein the form is a trihydrate.

7. 7. The crystalline form of claim 6, wherein the trihydrate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.4, 6.2, 11.6, 13.9, 16.4, and 16.7±0.2 degrees 2θ.

8. 7. The crystalline form of claim 6, wherein the trihydrate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 2】

9. 2. The crystalline form of claim 1, wherein the form is anhydrous.

10. 10. The crystalline form of claim 9, wherein the anhydrous form is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 6.2, 8.6, 9.7, 13.6, and 17.3±0.2 degrees 2θ.

11. 10. The crystalline form of claim 9, wherein the anhydrous form is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 3】

12. 10. The crystalline form of claim 9, wherein the anhydrous form is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 12.8, 13.6, 14.9, 16.1, and 17.2±0.2 degrees 2θ.

13. 10. The crystalline form of claim 9, wherein the anhydrous form is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 4】

14. 2. The crystalline form of claim 1, wherein the form is a solvate.

15. 15. The crystalline form of claim 14, wherein the form is a monoisopropanol solvate.

16. 16. The crystalline form of claim 15, wherein the monoisopropanol solvate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 7.2, 10.4, 10.8, 13.2, and 17.5±0.2 degrees 2θ.

17. 16. The crystalline form of claim 15, wherein the monoisopropanol solvate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 5】

18. 15. The crystalline form of claim 14, wherein the form is a tetraisopropanol solvate.

19. 20. The crystalline form of claim 18, wherein the tetraisopropanol solvate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.3, 6.4, 8.2, 10.5, 15.3, and 15.7±0.2 degrees 2θ.

20. 20. The crystalline form of claim 18, wherein the tetraisopropanol solvate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 6】

21. 2. The crystalline form of claim 1, wherein the form is a heterosolvate.

22. 22. The crystalline form of claim 21, wherein the form is a water-isopropanol heterosolvate.

23. 23. The crystalline form of claim 22, wherein the water-isopropanol heterosolvate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 7.3, 7.6, 10.4, 10.8, and 17.5±0.2 degrees 2θ.

24. 23. The crystalline form of claim 22, wherein the water-isopropanol heterosolvate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 7】

25. 15. The crystalline form of claim 14, wherein the form is a hexafluoroisopropanol solvate.

26. 26. The crystalline form of claim 25, wherein the hexafluoroisopropanol solvate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 4.3, 6.0, 6.9, 10.9, 11.5, 14.7, and 17.1±0.2 degrees 2θ.

27. 26. The crystalline form of claim 25, wherein the hexafluoroisopropanol solvate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 8】

28. 15. The crystalline form of claim 14, wherein the form is an ethanol solvate.

29. 30. The crystalline form of claim 28, wherein the ethanol solvate is characterized by one or more peaks in its X-ray powder diffraction pattern selected from 5.3, 10.6, and 15.9±0.2 degrees 2θ.

30. 30. The crystalline form of claim 28, wherein the ethanol solvate is characterized by the following peaks in its X-ray powder diffraction pattern: 【Table 9】

31. 31. A substantially impurity-free sample comprising the crystalline form of any one of claims 1 to 30.

32. 32. The sample of claim 31 comprising at least about 90% by weight of Compound 1.

33. 32. The sample of claim 31, comprising at least about 95% by weight of Compound 1.

34. 32. The sample of claim 31, comprising at least about 99% by weight of Compound 1.

35. 32. The sample of claim 31 containing less than about 5.0% total organic impurities.

36. 32. The sample of claim 31 containing less than about 3.0% total organic impurities.

37. 32. The sample of claim 31 containing less than about 1.5% total organic impurities.

38. 32. The sample of claim 31 containing less than about 1.0% total organic impurities.

39. 32. The sample of claim 31 containing less than about 0.5% total organic impurities.

40. 31. A method of inhibiting the activity of JAK2 kinase or a mutant thereof in a biological sample, comprising contacting said biological sample with a crystalline form or composition thereof according to any one of claims 1 to 30.

41. 31. A method of inhibiting activity of JAK2 kinase or a mutant thereof in a patient, comprising administering to said patient a crystalline form or composition thereof according to any one of claims 1-30.

42. 31. A method for treating a JAK2-mediated disease or disorder in a patient in need thereof, comprising administering to the patient a crystalline form of any one of claims 1-30, or a pharma- ceutically acceptable composition thereof.

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