Crystalline forms of RIP1 inhibitors

Abstract

Provided are crystalline forms of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, methods for preparing and characterizing the crystalline forms, and methods for using the crystalline forms to treat various diseases or conditions, such as diseases or conditions mediated by RIP1.

Description

[Technical field]

[0001] Related Applications This application claims priority to International Application No. PCT / CN2021 / 125895, filed October 22, 2021, the contents of which are incorporated by reference in their entirety herein.

[0002] The present disclosure relates to solid forms, e.g., crystalline forms, of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, methods of preparing and characterizing the crystalline forms, and methods of using the crystalline forms to treat a disease or condition, e.g., a disease or condition mediated by receptor-interacting protein 1 (RIP1). [Background technology]

[0003] Necroptosis is a highly regulated, caspase-independent form of cell death that is an important form of programmed cell death and plays a key role in many necrotic cell diseases that manifest in various pathological forms of cell death, including ischemic brain injury, neurodegenerative diseases, viral infections, and peripheral autoimmune diseases. (Dunai, et al., Dec 2011, Pathol. Oncol. Res.: POR 17(4):791-800. J. Med. Chem. 2020, 63, 4, 1490-1510. Nature Reviews Drug Discovery, 19, 553-571 (2020)). Tumor necrosis factor alpha (TNF-α)-induced NF-κB activation plays a central role in the immune system and inflammatory responses.

[0004] RIP1 is a multifunctional signal transduction agent involved in the activation of nuclear factor kappa B (NF-kappa B), apoptosis, and mediation of necroptosis. The kinase activity of RIP1 plays a key role in mediating necroptosis, a caspase-independent pathway of necrotic cell death (Holler et al. Nat Immunol 2000;1:489-495; Degterev et al. Nat Chem Biol 2008;4:313-321). RIP1 may contribute to D-1 immunotherapy resistance (e.g., Manguso et al., 2017 Nature 547,413-418) and act as a checkpoint kinase governing tumor immunity (e.g., Wang et al, Cancer Cell 34,757-774, Nov 12, 2018). RIP1 has emerged as a promising therapeutic target for the treatment of a wide range of human neurodegenerative, autoimmune, and inflammatory diseases, such as psoriasis, rheumatoid arthritis, and ulcerative colitis (Pharmacol. Res. Perspect. 2017, 5, e00365, PNAS May 14, 2019, 116(20)9714-9722), as well as central nervous system (CNS) indications such as amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (Nat. Rev. Neurosci. 2019, 20, 19-33).

[0005] Certain compounds for modulating necrosis or necroptosis are disclosed in U.S. Pat. No. 9,974,762, U.S. Pat. No. 10,092,529, U.S. Pat. No. 6,756,394, U.S. Pat. No. 8,278,344, U.S. Patent Application Publication No. 20120122889, U.S. Patent Application Publication No. 20090099242, U.S. Patent Application Publication No. 20100317701, U.S. Patent Application Publication No. No. 20110144169, U.S. Patent Application Publication No. 20030083386, U.S. Patent Application Publication No. 201200309795, WO 2009023272, WO 2010075290, WO 2010075561, WO 2012125544, WO 2020 / 103884, and WO 2020103859.

[0006] It is desirable to obtain various solid forms of RIP1 inhibitors, such as crystalline forms of the inhibitors, that are suitable for therapeutic applications and manufacturing processes. Summary of the Invention

[0007] One aspect of the disclosure provides a solid state, e.g., crystalline, form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide according to formula I: [ka]

[0008] In some embodiments, the disclosure provides, for example, crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide in substantially pure form.

[0009] In some embodiments, the disclosure provides, for example, crystalline Type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide in substantially pure form.

[0010] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a substantially pure crystalline form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (e.g., Type A or Type C) and a pharma- ceutically acceptable carrier. In some embodiments, the pharmaceutical composition may further comprise an additional active pharmaceutical agent.

[0011] Another aspect of the present disclosure provides a method of treating a disease or condition comprising administering a therapeutically effective amount of a substantially pure crystalline form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (e.g., Type A or Type C) or a pharmaceutical composition thereof to a subject in need thereof, wherein the disease or condition is selected from inflammatory diseases, immune diseases (e.g., autoimmune diseases), allergic diseases, transplant rejection, necrotic cell diseases, neurodegenerative diseases, CNS diseases, eye diseases, infectious diseases, malignancies, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and viral infections.

[0012] A further aspect of the present disclosure provides a method of treating a disease or condition mediated by RIP1, comprising administering to a subject in need thereof a therapeutically effective amount of a substantially pure crystalline form (e.g., Type A or Type C) of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, or a pharmaceutical composition thereof.

[0013] In some embodiments, the method of treatment comprises administering to a subject in need thereof an additional active pharmaceutical agent, either in the same pharmaceutical composition as the substantially pure crystalline form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (e.g., Type A or Type C), or in a separate composition. When administered as a separate dosage form, the additional therapeutic agent may be administered prior to, simultaneously with, or after administration of the crystalline form.

[0014] Also disclosed herein is a method of mediating, e.g., inhibiting, RIP1, comprising contacting a RIP1 protein or a fragment thereof with a substantially pure crystalline form (e.g., Type A or Type C) of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, or a pharmaceutical composition thereof.

[0015] In certain figures showing multiple graphs, a legend in a box in the upper right corner of the figure indicates, from top to bottom, the identification number of the material used in each graph in the figure. In certain figures, each graph is also separately annotated with the identification number of the material used in each graph. [Brief description of the drawings]

[0016] [Figure 1] FIG. 1 shows an XRPD pattern for crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (also referred to herein as “Type A Reference” or “Type A_817506-48-A”, Experiment ID 817506-48-A).

[0017] [Diagram 2] The DSC thermogram (bottom) and TGA graph (top) of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide are shown (Experiment ID 817506-48-A).

[0018] [Diagram 3] Proton NMR of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide using DMSO-d6 as the solvent (Experiment ID 817506-48-A).

[0019] [Figure 4] Shown is a PLM image of crystal type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (Experiment ID 817506-48-A).

[0020] [Figure 5A] FIG. 1 shows the XRPD pattern of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (non-micronized). [Figure 5B] FIG. 2 shows a DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (non-micronized), the first peak indicating melting of the compound and the second peak indicating decomposition of the compound.

[0021] [Figure 6A] FIG. 1 shows the XRPD pattern of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (microparticulated). [Figure 6B] Figure 2 shows the DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (micronized), where the first peak indicates melting of the compound and the second peak indicates decomposition of the compound. The particle size (D90) of the micronized sample was about 15 μm.

[0022] [Figure 7] FIG. 1 shows the XRPD patterns of two samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the antisolvent addition method (Experiment ID 810048-19-B1 and 810048-19-B2), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0023] [Figure 8] FIG. 1 shows a DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the antisolvent addition method (Experiment ID 810048-19-B2).

[0024] [Figure 9]FIG. 1 shows the XRPD patterns of five samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained by solid vapor diffusion (Experiment ID 810055-02-A1, Experiment ID 810055-02-A2, Experiment ID 810055-02-A3, Experiment ID 810055-02-A4, and Experiment ID 810055-02-A5), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0025] [Figure 10] FIG. 1 shows DSC thermograms of two samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from solid vapor diffusion method (Experiment ID 810055-02-A1 (top) and Experiment ID 810055-02-A3 (bottom)).

[0026] [Figure 11] FIG. 1 shows the XRPD patterns of four samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry by room temperature method (Experiment ID 810055-03-A1, Experiment ID 810055-03-A2, Experiment ID 810055-03-A3, and Experiment ID 810055-03-A4), a Type A standard (Experiment ID 817506-48-A), and an impurity B standard.

[0027] [Figure 12] FIG. 1 shows the XRPD patterns of four samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry by room temperature method (Experiment ID 810055-03-A5, Experiment ID 810055-03-A6, Experiment ID 810055-03-A7, and Experiment ID 810055-03-A8), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0028] [Figure 13]FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry by room temperature method (Experiment ID 810055-03-A9, Experiment ID 810055-03-A10, and Experiment ID 810055-03-A11), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0029] [Figure 14] FIG. 1 shows DSC thermograms of three samples of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from slurries by the room temperature method (Experiment ID 810055-03-A1 (top), Experiment ID 810055-03-A6 (middle), Experiment ID 810055-03-A9 (bottom)).

[0030] [Figure 15] FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry using the 50° C. Method (Experiment ID 810055-04-A1, Experiment ID 810055-04-A2, and Experiment ID 810055-04-A3), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0031] [Figure 16] FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from slurries using the 50° C. Method (Experiment ID 810055-04-A5, Experiment ID 810055-04-A6, and Experiment ID 810055-04-A7), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0032] [Figure 17]FIG. 1 shows DSC thermograms of two samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry by the 50° C. Method (Experiment ID 810055-04-A3 (top) and Experiment ID 810055-04-A6 (bottom)).

[0033] [Figure 18] FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the slow evaporation method (Experiment ID 810055-05-A1, Experiment ID 810055-05-A2, and Experiment ID 810048-19-B3), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0034] [Figure 19] FIG. 1 shows DSC thermograms of three samples of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the slow evaporation method (Experiment ID 810055-05-A1, Experiment ID 810055-05-A2, and Experiment ID 810048-19-B3).

[0035] [Figure 20] FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the slow cooling method (Experiment ID 810055-06-A1 (top), Experiment ID 810055-06-A2 (middle), and Experiment ID 810048-19-B5 (bottom)), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0036] [Figure 21] FIG. 1 shows DSC thermograms of two samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the slow cooling method (Experiment ID 810055-06-A2 (top) and Experiment ID 810048-19-B5 (bottom)).

[0037] [Figure 22] FIG. 1 shows the XRPD patterns of four samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained by liquid vapor diffusion (Experiment ID 810055-07-A1, Experiment ID 810055-07-A2, Experiment ID 810055-07-A3, and Experiment ID 810048-19-B6), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0038] [Diagram 23] FIG. 1 shows DSC thermograms of three samples of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from liquid vapor diffusion method (Experiment ID 810055-07-A1 (top), Experiment ID 810055-07-A3 (middle), and Experiment ID 810048-19-B6 (bottom)).

[0039] [Figure 24] FIG. 1 shows the XRPD patterns of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from a slurry by the grinding method (Experiment ID 810055-08-A1, Experiment ID 810055-08-A2, and Experiment ID 810055-08-A3), a Type A reference (Experiment ID 817506-48-A), and an Impurity B reference.

[0040] [Diagram 25] FIG. 1 shows DSC thermograms of three samples of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from the grinding method (Experiment ID 810055-08-A2 (top), Experiment ID 810055-08-A1 (middle), and Experiment ID 810055-08-A3 (bottom)).

[0041] [Figure 26]FIG. 1 shows XRPD patterns of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from MeOH / HO (Experimental ID 817506-48-A2) and type A standard (Experimental ID 817506-48-A).

[0042] [Figure 27] 1 shows a DSC thermogram (bottom) and a TGA graph (top) of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from MeOH / H2O (experimental ID 817506-48-A2).

[0043] [Figure 28] FIG. 1 shows a PLM image of crystals type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from MeOH / HO (experimental ID 817506-48-A2).

[0044] [Figure 29] FIG. 1 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from acetone / HO (Experimental ID 817506-48-A1) and EtOAc / n-heptane (Experimental ID 817506-48-A4), as well as a Type A standard (Experimental ID 817506-48-A).

[0045] [Diagram 30] FIG. 1 shows the XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained from MeOH / HO (Experimental ID 810048-03-A1) as well as Type A standard (Experimental ID 817506-48-A) and Impurity B standard.

[0046] [Diagram 31]FIG. 1 shows a DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide obtained in MeOH / HO (experimental ID 810048-03-A1).

[0047] [Diagram 32] 1 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide before (Experiment ID 817506-48-A2) and after (Experiment ID 817506-50-A) solubility testing, and of Type A standard (Experiment ID 817506-48-A). [Diagram 33] HPLC chromatograms of type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (817506-48-A2) for stability evaluation: (1): blank; (2): 80°C / 1 day; (3): 25°C / 60% RH / 1 week; (4): 40°C / 75% RH / 1 week.

[0048] [Diagram 34] FIG. 1 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after stability studies, Experiment IDs 810048-01-A1 (80° C. / 1 day), 810048-01-A2 (25° C. / 60% RH / 1 week), and 810048-01-A3 (40° C. / 75% RH / 1 week), as well as a Type A standard (Experiment ID 817506-48-A). [Diagram 35] 1 shows a DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after storage at 80° C. (Experiment ID 817506-48-A2).

[0049] [Diagram 36] FIG. 1 shows a DSC thermogram of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after storage at 25° C. / 60% RH (Experiment ID 817506-48-A2).

[0050] [Figure 37] Figure 2 shows a DSC thermogram of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after storage at 40°C / 75% RH (Experiment ID 817506-48-A2).

[0051] [Figure 38] The HPLC chromatograms of Type A (817506-48-A2) after stability testing under white light conditions are shown. (1): Blank; (2) White light 1,200,000 Lux·hrs; (3): White light_control.

[0052] [Figure 39] The HPLC chromatograms of Type A (817506-48-A2) after stability testing under UV conditions are shown. (1): Blank; (2): UV200W·hrs / m2; (3): UV_control.

[0053] [Diagram 40] FIG. 1 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, Type A standard (Experimental ID 817506-48-A), and Impurity B standard after stability testing under white light conditions, Experimental ID 817506-48-A2 (white light), 817506-48-A2 (white light_control).

[0054] [Diagram 41] Figure 2 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, Type A standard (Experimental ID 817506-48-A), and Impurity B standard after stability testing under UV conditions, Experimental ID 817506-48-A2 (UV), 817506-48-A2 (UV_Control).

[0055] [Diagram 42]FIG. 1 shows DSC thermograms of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after stability testing under white light conditions or under white light control.

[0056] [Diagram 43] FIG. 1 shows a DSC thermogram of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide under UV conditions or as a UV control.

[0057] [Diagram 44] The DVS graph of Type A (817506-48-A2) is shown.

[0058] [Diagram 45] 1 shows XRPD patterns of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide before and after DVS testing, along with a Type A standard.

[0059] [Figure 46] 1 shows a DSC thermogram of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide after DVS testing.

[0060] [Figure 47] The XRPD pattern of crystalline Type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide is shown along with a Type A standard.

[0061] [Figure 48] The XRPD patterns of crystalline Type C N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide before and after drying are shown along with a Type A standard.

[0062] [Figure 49]Figure 1 shows the XRPD patterns of three samples of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (SN-045-17, micronized YF2022-2-API, and micronized YF2022-3-API). The particle size (D90) of the "micronized YF2022-2-API" and "micronized YF2022-3-API" samples was estimated to be approximately 30-40 μm.

[0063] [Figure 50] The XRPD pattern of one sample of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (810048-18-B) is shown along with a Type A standard.

[0064] [Figure 51] Shown is a DSC thermogram (bottom) and a TGA graph (top) of one sample (810048-18-B) of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide.

[0065] [Figure 52] The XRPD pattern of one sample of crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (810055-01-A) is shown along with a Type A standard.

[0066] [Figure 53] Shown is a DSC thermogram (bottom) and a TGA graph (top) of one sample (810055-01-A) of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide.

[0067] [Figure 54] The proton NMR of one sample (810055-01-A) of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide is shown.

[0068] [Figure 55] 1 shows the XRPD patterns of four samples of crystalline Type A and Impurity B of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, along with a Type A standard and an Impurity B standard.

[0069] [Figure 56] 1 shows DSC thermograms of four samples of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide and impurity B. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0070] All publications, including but not limited to patents and patent applications, cited herein are hereby incorporated by reference as if fully set forth. In the event that any particular content of a reference cited herein contradicts or is inconsistent with the present disclosure, the present disclosure shall control.

[0071] Any one embodiment of the present disclosure described herein, including embodiments described only in a section of this specification describing particular aspects of the disclosure and embodiments described only in the examples or drawings, may be combined with any other one or more embodiments, unless expressly prohibited or inappropriate.

[0072] I. Definition It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0073] Although any methods and materials similar or equivalent to those described herein can be used in the practice of testing of the present disclosure, exemplary materials and methods are described herein. In describing and claiming the present disclosure, the following terminology is used.

[0074] Abbreviations for certain solvents described herein are provided below. [Table 1]

[0075] As used herein, the singular forms "a," "an," and "the" include plural references unless the context dictates otherwise. Thus, for example, reference to "a compound" includes a combination of two or more compounds, and the like.

[0076] The term "about" as used herein means having a value that falls within the mean tolerance criteria when considered by one of ordinary skill in the art, for example, ±20%, preferably ±10%, more preferably ±5%, or even more preferably ±2% of the mean.

[0077] The term "substantially the same" or "substantially as represented" means that typical variability for a particular method is taken into account. For example, with respect to peak positions in the context of XRPD, DSC, TGA, DVS, and NMR, the term "substantially the same" or "substantially as represented" means that typical variability in peak positions and intensities is taken into account. Those skilled in the art will understand that peak positions will show some variability. Furthermore, those skilled in the art will understand that relative peak intensities will show variability between instruments, as well as variability due to degree of crystallization, particle size, preferred orientation, sample surface prepared, and other factors known to those skilled in the art.

[0078] The terms "substantially pure" or "substantially free" with respect to a particular crystalline form of a compound means that a composition comprising the crystalline form contains less than 30%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.2%, or less than 0.1% by weight of other substances, including other crystalline or other solid state forms, and / or impurities. In some embodiments, "substantially pure" or "substantially free" refers to a material that is free of other substances, including other crystalline forms, other solid state forms, and / or impurities. Impurities may include, for example, by-products or residual reagents from chemical reactions, contaminants, decomposition products, other crystalline forms, water, and solvents.

[0079] In some embodiments, compositions comprising crystalline forms of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide may also include impurities, such as impurity B, substantially as shown in certain figures herein, such as FIG. 7. The impurity, such as impurity B, may be present in the compositions disclosed herein in an amount of less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt%, less than 0.5 wt%, less than 0.2 wt%, or less than 0.1 wt%. In some embodiments, impurity B may be present in the compositions disclosed herein in an amount of about 0.1 wt% to 0.5 wt%, such as about 0.14 wt%.

[0080] The term "crystalline" as used herein means having a regularly repeating arrangement of molecules or external face planes. Crystal forms can differ with respect to thermodynamic stability, physical parameters, X-ray structure, and preparation method.

[0081] The term "micronization" as used herein refers to a process that reduces the average diameter of particles of a solid material to the micrometer range, or even to the nanometer scale. Micronization processes may utilize mechanical means such as milling and grinding, or utilize the properties of supercritical fluids and manipulate the principles of solubility. A "micronized" material means that the material has undergone a particular micronization process to reduce particle size.

[0082] As used herein, "API" means "active pharmaceutical ingredient," such as (3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide, for example, crystalline Type A of such compound.

[0083] The term "subject" refers to animals, including humans.

[0084] As used herein, the term "pharmaceutical acceptable" refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response, and the like, and is commensurate with a reasonable benefit / risk ratio.

[0085] The term "therapeutically effective amount" refers to an amount of a compound, e.g., a crystalline form of Formula I disclosed herein, that when administered results in a desired effect (e.g., amelioration of a disease or condition, reduction in the severity of a disease or condition, and / or reduction in the progression of a disease or condition), wherein the disease or condition is selected from an inflammatory disease, an immune disease (e.g., an autoimmune disease), an allergic disease, a transplant rejection, a necrotic cell disease (e.g., a disease associated with necroptosis), a neurodegenerative disease, a CNS disease, an ischemic brain injury, an eye disease, an infectious disease, and a malignancy (including those mediated by receptor interacting protein 1 (RIP1) signaling); a disease or condition selected from an ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and a viral infection (including those mediated by RIP1 signaling); a disease mediated by RIP1 signaling. The precise amount of a therapeutically effective amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lloyd (1999), The Art, Science and Technology of Pharmaceutical Compounding).

[0086] As used herein, the term "treatment" and its synonyms refer to slowing or stopping disease progression. As used herein, "treatment" and its synonyms include, but are not limited to, complete or partial remission of a disease or condition or its symptoms, cure of a disease or condition or its symptoms, and reducing the risk of a disease or condition, including a disease or condition selected from inflammatory diseases, immune diseases (e.g., autoimmune diseases), allergic diseases, graft rejection, necrotic cell diseases, neurodegenerative diseases, CNS diseases, ischemic brain injury, eye diseases, infectious diseases, and malignant tumors, including those mediated by receptor interacting protein 1 (RIP1) signaling; a disease or condition selected from ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and viral infections, including those mediated by receptor interacting protein 1 (RIP1) signaling; a disease or condition mediated by RIP1 signaling. Improvement or reduction in the severity of any of these symptoms can be assessed according to methods and techniques known in the art.

[0087] As used herein, the term "N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide" or "Formula I" refers to a compound, or one or more variations of the compound, such as tautomers, solvates (e.g., hydrates), and pharma- ceutically acceptable salts of the foregoing. Compounds, tautomers, solvates (e.g., hydrates), and pharma-ceutically acceptable salts may also be used, for example, with -CD in place of methyl. 3 , CD 2 H or CDH 2 Such compounds may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds, such as deuterium. For example, the compounds may contain radioactive isotopes, such as tritium ( 3 H) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are intended to be encompassed within the scope of the present disclosure.

[0088] II.Crystal form One aspect of the disclosure provides a solid form, such as a crystalline or amorphous form, of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (Formula I).

[0089] In some embodiments, the present disclosure provides crystalline Type A of formula I, eg, crystalline Type A substantially free of other solid forms of formula I.

[0090] In some embodiments, the present disclosure provides crystalline Type C of formula I, eg, crystalline Type C that is substantially free of other solid forms of formula I.

[0091] In some embodiments, the present disclosure provides an amorphous form of formula I, eg, an amorphous form that is substantially free of other solid forms of formula I.

[0092] In some embodiments, the disclosure provides a composition comprising crystalline Type A of formula I. In some embodiments, the disclosure provides a composition comprising crystalline Type C of formula I.

[0093] In some embodiments, the present disclosure provides a composition comprising crystalline Type A of Formula I and an amorphous form.

[0094] In some embodiments, the present disclosure provides a composition comprising crystalline Type A and crystalline Type C of Formula I.

[0095] In some embodiments, the present disclosure provides a composition comprising crystalline Type A, crystalline Type C, and an amorphous form of Formula I.

[0096] Techniques for characterizing the crystalline forms of the present disclosure include, but are not limited to, powder X-ray diffractometry (XRPD), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), dynamic vapor sorption (DVS), polarized light microscopy (PLM), vibrational spectroscopy (e.g., IR spectroscopy and Raman spectroscopy), scanning electron microscopy, solid-state NMR (nuclear magnetic resonance), hot stage optical microscopy, electron crystallography, single crystal X-ray diffraction, quantitative analysis, particle size analysis (e.g., particle size, particle size distribution, and particle shape), specific surface area analysis, surface energy analysis (e.g., inverse gas chromatography or IGC), solubility and dissolution tests, or combinations of these techniques.

[0097] In some embodiments, the crystalline form is characterized by an X-ray powder diffraction pattern (XRPD). The XRPD diffractogram is typically represented by a plot of the intensity of the peaks versus the position of the peaks, i.e., the diffraction angle 2θ (2 theta) degrees. Characteristic peaks of a given XRPD can be selected according to the peak positions and their relative intensities to distinguish this crystalline structure from other crystalline structures. Those skilled in the art will recognize that the measurements of XRPD peak positions and / or intensities for a given crystalline form of the same compound will vary as disclosed herein. The value of 2θ degrees allows for appropriate variation.

[0098] Differences in XRPD patterns between separate measurements of the same polymorph can occur for many reasons. Sources of variation include sample preparation variation (e.g., sample height), instrument variation, particle size variation, calibration variation, and operator variation (including variation in determining peak positions). Lack of preferred orientation, i.e., random orientation of crystals in an XRPD sample, can result in significant differences in relative peak heights. Particle size can also cause significant differences in relative peak heights. For example, generally, the smaller the particle size, the stronger the signal of the diffraction peaks. Calibration and sample height errors often result in all shifts of the diffractogram peaks in the same direction and by the same amount. Small differences in sample height on a flat holder can result in large displacements of the XRPD peak positions. See Chen et al., J. Pharmaceutical and Biomedical Analysis 30 (2001) 26:63 for a systematic study showing that a sample height difference of 1 mm can result in peak shifts as high as 1-28.

[0099] Typically, the tolerance is expressed as "±". For example, "(±0.2 degrees 2θ at 8.7)" means "at about 8.7±0.2 degrees 2θ", indicating a range from about (8.7+0.2), i.e. 8.9, to about (8.7-0.2), i.e. about 8.5. Depending on the sample preparation technique, the calibration technique applied to the device, human operational variation, etc., one of skill in the art will recognize that an appropriate tolerance for XRPD may be ±0.5; ±0.4; ±0.3; ±0.2; ±0.1; ±0.05; or less. In some embodiments of the present disclosure, the XRPD tolerance is ±0.2. In some embodiments of the present disclosure, the XRPD tolerance is ±0.5.

[0100] In many cases, peak shifts between diffraction patterns due to systematic errors can be eliminated by compensating for the shifts (e.g., applying a correction factor to all peak position values) or by recalibrating the diffractometer. In general, the same techniques can be used to compensate for differences between diffractometers so that XRPD peak positions obtained from two different instruments can be made to match. Furthermore, when these techniques are applied to XRPD measurements from the same or different diffractometers, the peak positions of a particular polymorph will usually match to within about ±0.2.

[0101] Type A of Formula I In some embodiments, crystalline Type A of Formula I has an XRPD pattern substantially identical to one of the XRPD patterns displayed for crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 1, 5A, 6A, 7, 9, 11, 12, 13, 15, 16, 18, 20, 22, 24, 26, 29, 30, 32, 34, 40, 41, 45, 47, 48, 49, 50, 52, and 55.

[0102] In some embodiments, crystalline Type A of Formula I has an XRPD pattern characterized by at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, or at least nine of the 2θ peaks having maximum intensity substantially as shown in one of the XRPD patterns displayed for crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 1, 5A, 6A, 7, 9, 11, 12, 13, 15, 16, 18, 20, 22, 24, 26, 29, 30, 32, 34, 40, 41, 45, 47, 48, 49, 50, 52, and 55.

[0103] In some embodiments, crystalline Type A of Formula I has an XRPD pattern characterized by at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or at least eleven of the 2θ peaks having the greatest intensities substantially as shown in the XRPD pattern of FIG. 5A.

[0104] In some embodiments, crystalline Type A of Formula I has an XRPD pattern characterized by at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or at least eleven of the 2θ peaks having the greatest intensities substantially as shown in the XRPD pattern of FIG. 6A.

[0105] In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes a peak at 13.96 degrees 2θ (±0.2 degrees 2θ) and one of the following peaks at 9.15, 14.15, 17.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes a peak at 13.96 degrees 2θ (±0.2 degrees 2θ) and two of the following peaks at 9.15, 14.15, 17.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 13.96 and 14.15 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern comprising peaks at any three degrees 2θ (±0.2 degrees 2θ) selected from the group consisting of 9.15, 13.96, 14.15, 17.15, 18.22, and 26.31.

[0106] In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes a peak at 18.20 degrees 2θ (±0.2 degrees 2θ) and one of the following peaks at 9.10, 17.10, 21.7, and 27.40 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes a peak at 18.20 degrees 2θ (±0.2 degrees 2θ) and two of the following peaks at 9.10, 17.10, 21.7, and 27.40 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.10 and 18.20 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern comprising peaks at any three degrees 2θ (±0.2 degrees 2θ) selected from the group consisting of 9.10, 17.10, 18.20, 21.7, and 27.40.

[0107] In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 17.15, and 18.22 degrees 2θ (2 theta) (± 0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 17.15, 18.22, 21.8, and 27.40 degrees 2θ (2 theta) (± 0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 13.34, 17.15, 18.22, 21.80, and 27.40 degrees 2θ (2 theta) (± 0.2 degrees 2θ).

[0108] In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 13.96, 14.15, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 13.96, 14.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 13.96, 14.15, 17.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 13.96, 14.15, 17.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 13.96, 14.15, 17.15, 18.22, 20.38, 26.31, and 27.40 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.15, 13.34, 13.96, 14.15, 17.15, 18.22, 20.38, 20.93, 21.80, 21.93, 26.31, and 27.40 degrees 2θ (±0.2 degrees 2θ).

[0109] In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.10, 17.10, and 18.20 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.10, 17.10, 18.20, and 21.7 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type A of Formula I has an XRPD pattern that includes peaks at 9.10, 17.10, 18.20, 21.7, and 27.40 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Formula I Form I has an XRPD pattern that includes peaks at 9.10, 13.9, 17.10, 18.20, 21.7, and 27.40 degrees 2θ (±0.2 degrees 2θ).

[0110] In one embodiment, crystalline Type A of Formula I comprises XRPD peaks substantially as set forth in Table 1 below. The XRPD peaks set forth in Table 1 were obtained from a crystalline sample having a particle size (D90) of about 15 μm. [Table 2-1] [Table 2-2]

[0111] In one embodiment, crystalline Type A of Formula I comprises XRPD peaks substantially as shown in Table 2 below. [Table 3]

[0112] In some embodiments, the crystalline type A of formula I may exhibit, prior to decomposition of the compound, a DSC thermogram substantially similar to one of the DSC thermograms shown in Figure 2, Figure 5B (prior to decomposition of the compound), Figure 6B (prior to decomposition of the compound), Figure 8, Figure 10, Figure 14, Figure 17, Figure 19, Figure 21, Figure 23, Figure 25, Figure 27, Figure 31, Figure 35, Figure 36, Figure 37, Figure 42, Figure 43, Figure 46, Figure 51, Figure 53, and Figure 56. In certain embodiments, the crystalline type A has an onset melting temperature of about 94°C to about 96°C. In certain embodiments, the crystalline type A has an onset melting temperature of about 94.5°C to about 95.5°C. In certain embodiments, the crystalline type A has an onset melting temperature of about 95°C. In certain embodiments, the crystalline type A has an onset melting temperature of about 94.8°C. In one particular embodiment, crystalline Type A has an onset melting temperature of about 95.1°C.

[0113] In some embodiments, crystalline Type A of Formula I may exhibit a TGA graph substantially similar to one of the TGA graphs shown in FIG. 3, FIG. 27, FIG. 51, and FIG.

[0114] In some embodiments, crystalline Type A of Formula I may exhibit a DVS graph substantially similar to one of the DVS graphs shown in FIG.

[0115] In some embodiments of Crystal Type A of Formula I, at least one, at least two, at least three, or all of the following (a)-(d) apply to Crystal Type A of Formula I: (a) among the XRPD patterns displayed for Crystal Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 1, 5A, 6A, 7, 9, 11, 12, 13, 15, 16, 18, 20, 22, 24, 26, 29, 30, 32, 34, 40, 41, 45, 47, 48, 49, 50, 52, and 55, 2, 5B (before decomposition of the compound), 6B (before decomposition of the compound), 8, 10, 14, 17, 19, 21, 23, 25, 27, 31, 35, 36, 37, 42, 43, 46, 51, 53, and 56, prior to decomposition of the compound; (c) a TGA graph substantially the same as one of the TGA graphs shown in Figures 3, 27, 51, and 53; and (d) a DVS graph substantially the same as one of the DVS graphs shown in Figure 44.

[0116] In some embodiments, crystalline Type A of Formula I has the following characteristics: (a) an XRPD pattern substantially the same as the XRPD pattern shown in FIG. 1; (b) a DSC thermogram substantially the same as the DSC thermogram shown in FIG. 2; and (c) a TGA graph substantially the same as the TGA graph shown in FIG. 2.

[0117] In some embodiments, crystalline Type A of Formula I has the following characteristics: (a) an XRPD pattern substantially the same as the XRPD pattern shown in FIG. 6A; and (b) a DSC thermogram substantially the same as the DSC thermogram shown in FIG. 6B.

[0118] In some embodiments, the crystalline Type A of Formula I is anhydrous.

[0119] In some embodiments, crystalline Type A of Formula I is non-hygroscopic. In certain embodiments, dynamic vapor sorption (DVS) data for crystalline Type A of Formula I shows water uptake of about 0.10% at 80% RH / 25° C.

[0120] In some embodiments, H at room temperature 2 The equilibrium solubility of crystalline Type A of Formula I in O is approximately 0.15 mg / mL.

[0121] Crystalline Type A of Formula I is substantially stable. In certain embodiments, no change in crystal type is detected after suspending crystalline Type A of Formula I in HO for 24 hours. In certain embodiments, the solid-state stability of Type A is evaluated for 1 day at 80° C., 1 week at 25° C. / 60% RH, and 1 week at 40° C. / 75% RH. Under these conditions, no change in crystal morphology or loss in HPLC purity is detected. In certain embodiments, the photostability of Type A is evaluated under white light (1,200,000 Lux·hrs) and UV (200 W·hrs / m 2 ) is evaluated under both conditions. Under either condition, no change in crystal morphology or loss in HPLC purity is observed.

[0122] As further shown in the examples, crystalline Type A of Formula I can be prepared by methods such as antisolvent addition, solid vapor diffusion, solution vapor diffusion, slurrying, slow evaporation, slow cooling, polymer-induced crystallization, and milling.

[0123] In some embodiments, the crystalline Type A of Formula I is: dissolving N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide in a first solvent to obtain a solution; adding a second solvent to the solution while stirring the solution to obtain a suspension; isolating the solid from the suspension; drying the solid to obtain crystalline type A; The composition may be produced by a method comprising:

[0124] In certain embodiments, the first solvent is a solvent in which N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide has a solubility of greater than 10 mg / mL, such as greater than 15 mg / mL, greater than 20 mg / mL, greater than 25 mg / mL, greater than 30 mg / mL, or greater than 35 mg / mL; and the second solvent is a solvent in which N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide has a solubility of less than 10 mg / mL, such as less than 8 mg / mL, less than 6 mg / mL, less than 4 mg / mL, less than 3 mg / mL, less than 2 mg / mL, or less than 1 mg / mL.

[0125] In certain embodiments, the first solvent is isoamyl alcohol, ethyl lactate, MEK, anisole, n-butanol / n-BuOH, ethyl formate, 2,2,2-trifluoroethanol, toluene, pyridine, isobutanol, chlorobenzene, CPME, m-xylene, n-butyl acetate, cumene, NMP, MTBE, 2-MeTHF, EtOAc, acetone, THF, DMAc, IPA, EtOH, DCM, MeOH, IPA, MIBK, IPAc, THF, 1,4-dioxane, DCM, CHCl 3 , toluene, DMSO, DMAc, NMP, and ACN, and the second solvent is selected from the group consisting of n-hexane, water, 2-MeTHF, MTBE, cyclohexane, and n-heptane.

[0126] In certain embodiments, the first solvent is selected from the group consisting of EtOAc, acetone, THF, DMAc, IPA, EtOH, DCM, and MeOH; and the second solvent is selected from the group consisting of water, 2-MeTHF, cyclohexane, and n-heptane.

[0127] Type C of Formula I In some embodiments, crystalline Type C of Formula I has an XRPD pattern substantially identical to one of the XRPD patterns displayed for crystalline Type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 47 and 48.

[0128] In some embodiments, crystalline Type C of Formula I has an XRPD pattern characterized by at least two, at least three, at least four, at least five, or at least six of the 2θ peaks having the greatest intensity substantially as shown in one of the XRPD patterns displayed for crystalline Type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 47 and 48.

[0129] Crystalline type C of formula I can be converted to type A after drying.

[0130] In some embodiments, crystalline Type C of Formula I has an XRPD pattern including a peak at 21.78 degrees 2θ (±0.2 degrees 2θ) and one of the following peaks at 9.08, 13.45, 18.18, and 27.39 degrees 2θ (±0.2 degrees 2θ).

[0131] In some embodiments, crystalline Type C of Formula I has an XRPD pattern including a peak at 21.78 degrees 2θ (±0.2 degrees 2θ) and two of the following peaks at 9.08, 13.45, 14.28, 18.18, and 27.39 degrees 2θ (±0.2 degrees 2θ).

[0132] In some embodiments, crystalline Type C of Formula I has an XRPD pattern including a peak at 21.78 degrees 2θ (±0.2 degrees 2θ) and three of the following peaks at 9.08, 13.45, 14.28, 18.18, and 27.39 degrees 2θ (±0.2 degrees 2θ).

[0133] In some embodiments, crystalline Type C of Formula I has an XRPD pattern that includes peaks at 9.08, 18.18, and 21.78 degrees 2θ (2 theta) (± 0.2 degrees 2θ). In some embodiments, crystalline Type C of Formula I has an XRPD pattern that includes peaks at 9.08, 18.18, 21.78, and 27.39 degrees 2θ (± 0.2 degrees 2θ). In some embodiments, crystalline Type C of Formula I has an XRPD pattern that includes peaks at 9.08, 13.45, 14.28, 18.18, 21.78, and 27.39 degrees 2θ (± 0.2 degrees 2θ). In some embodiments, crystalline Type C of Formula I has an XRPD pattern that includes peaks at 9.08, 13.45, 13.91, 14.28, 18.18, 21.78, 27.39, 30.80, 36.77, and 37.14 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Type C of Formula I has an XRPD pattern that includes peaks at any three degrees 2θ (±0.2 degrees 2θ) selected from the group consisting of 9.08, 13.45, 14.28, 18.18, 21.78, and 27.39.

[0134] In one embodiment, crystalline Type C of Formula I comprises XRPD peaks substantially as shown in Table 3 below. [Table 4]

[0135] III. Composition Another aspect of the disclosure provides pharmaceutical compositions comprising a crystalline form of Formula I (e.g., Type A or Type C) and at least one pharma- ceutically acceptable carrier. In some embodiments, the crystalline form of Formula I (e.g., Type A or Type C) in the pharmaceutical composition is substantially free of other solid forms of Formula I.

[0136] In some embodiments, the pharma- ceutically acceptable carrier is selected from a pharma- ceutically acceptable vehicle and a pharma- ceutically acceptable adjuvant, hi some embodiments, the pharma- ceutically acceptable carrier is selected from a pharma- ceutically acceptable filler, disintegrant, surfactant, binder, and lubricant.

[0137] It will also be understood that the pharmaceutical compositions of the present disclosure can be used in combination therapy: i.e., the pharmaceutical compositions described herein can further comprise an additional active pharmaceutical agent. Alternatively, a pharmaceutical composition comprising a substantially pure crystalline form of Formula I (e.g., Type A or Type C) can be administered as a separate composition, simultaneously with a composition comprising the additional active pharmaceutical agent, prior to a composition comprising the additional active pharmaceutical agent, or after a composition comprising the additional active pharmaceutical agent.

[0138] In some embodiments, pharma- ceutically acceptable carriers can be selected from adjuvants and vehicles. Pharmaceutically acceptable carriers used herein can be selected from, for example, any and all solvents, diluents, other liquid vehicles, dispersants, suspending agents, surfactants, isotonicity agents, thickening agents, emulsifiers, preservatives, solid binders, and lubricants suitable for the specific dosage form desired. Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. DB Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 1988 to 1999, Marcel Dekker, New York disclose various carriers used in formulating pharmaceutical compositions and known techniques for their preparation. Except insofar as any conventional carrier is incompatible with the compounds of the present disclosure, for example, by producing any undesirable biological effects or by otherwise interacting in a deleterious manner with any other components of the pharmaceutical composition, its use is contemplated within the scope of the present disclosure.Non-limiting examples of suitable pharma- ceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates, glycine, sorbic acid, and potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts, and electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, wool fat, sugars (such as lactose, glucose, and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as carboxymethylcellulose, cellulose acetate, cellulose esters ... Ingredients that can be used include, for example, sodium cellulose, ethyl cellulose, and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository wax), oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (such as propylene glycol and polyethylene glycol), esters (such as ethyl oleate and ethyl laurate), agar, buffers (such as magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer, non-toxic compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), colorants, releasing agents, coating agents, sweeteners, flavors, fragrances, preservatives, and antioxidants.

[0139] The substantially pure crystalline forms (e.g., Type A or Type C) or pharmaceutical compositions of formula I disclosed herein can be administered orally in solid dosage forms, such as capsules, tablets, troches, dragees, granules, and powders, or in liquid dosage forms, such as elixirs, syrups, emulsions, dispersions, and suspensions. The crystalline forms of formula I described herein can also be administered parenterally in sterile liquid dosage forms, such as dispersions, suspensions, or solutions. Other dosage forms that can also be used to administer the crystalline forms of formula I described herein are as ointments, creams, drops, transdermal patches, or powders for topical administration, ophthalmic solution or suspension formulations for ocular administration, such as eye drops, as aerosol sprays or powder compositions for inhalation or intranasal administration, or as creams, ointments, sprays, or suppositories for rectal or vaginal administration.

[0140] Gelatin capsules containing the crystalline form of formula I disclosed herein and powdered carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid can also be used.Compressed tablets can be manufactured using similar diluents.Both tablets and capsules can be manufactured as sustained release products, and can release medicine continuously for a certain period of time.Compressed tablets can be sugar-coated or film-coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric-coated to selectively disintegrate in the gastrointestinal tract.

[0141] Liquid dosage forms for oral administration may further comprise at least one agent selected from coloring and flavoring agents to enhance patient acceptance.

[0142] Generally, water, suitable oil, saline, aqueous dextrose (glucose) solution and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol may be examples of suitable carriers for parenteral solutions. Solutions for parenteral administration may contain a water-soluble salt of at least one compound described herein, at least one suitable stabilizer, and at least one buffer substance, if necessary. Antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or in combination, may be examples of suitable stabilizers. Citric acid and its salts and sodium EDTA may also be used as examples of suitable stabilizers. In addition, parenteral solutions may further contain at least one preservative (antiseptic), for example, selected from benzalkonium chloride, methyl and propyl paraben, and chlorobutanol.

[0143] Pharmaceutically acceptable carriers are selected from carriers that are compatible with the active ingredients of the composition (in some embodiments, can stabilize the active ingredients) and are not harmful to the subject being treated. For example, solubilizers such as cyclodextrins (which can form specific and more soluble complexes with at least one compound and / or at least one pharma-ceutically acceptable salt disclosed herein) can be utilized as pharmaceutical excipients for delivery of active ingredients. Other examples of carriers include colloidal silicon dioxide, magnesium stearate, cellulose, sodium lauryl sulfate, and pigments such as D&C Yellow#10. Suitable pharma-ceutically acceptable carriers are described in Remington's Pharmaceutical Sciences, A. Osol.

[0144] For administration by inhalation, the crystalline form of formula I described herein can be conveniently delivered in the form of aerosol spray from a pressurized pack or nebulizer.The crystalline form of formula I described herein can also be delivered as a powder that can be formulated, and the powder composition can be inhaled using an insufflation powder inhaler.One exemplary delivery system for inhalation can be metered dose inhalation (MDI) aerosol, which can be formulated as a suspension or solution of the crystalline form described herein in at least one suitable propellant selected from, for example, fluorocarbons and hydrocarbons.

[0145] For ocular administration, ophthalmic preparations can be formulated with a solution or suspension of the crystalline form of Formula I described herein in an appropriate weight percentage in a suitable ophthalmic vehicle such that the crystalline form of Formula I described herein is maintained in contact with the ocular surface for a period of time sufficient to allow the active compound to penetrate the corneal and interior regions of the eye.

[0146] Pharmaceutical dosage forms useful for administration of the crystalline forms of Formula I described herein include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injections, and oral suspensions. In some embodiments, the pharmaceutical compositions disclosed herein may be in the form of controlled release or sustained release compositions known in the art.

[0147] The term "unit dosage form" refers to physically discrete units suitable as unitary dosage forms for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampoules or syringes in the case of liquid compositions, or pills, tablets, capsules, drops, and the like in the case of solid compositions. In such compositions, the active material is usually in the range of about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight, with the remainder being various vehicles or carriers and processing aids that serve to form the desired dosage form. The unit dosage formulations are preferably about 5, 10, 25, 50, 100, 250, 500, or 1,000 mg per unit. In certain embodiments, the unit dosage forms are packaged in multipacks adapted for sequential use, such as blister packs containing at least 6, 9, or 12 sheets of unit dosage forms.

[0148] In some embodiments, unit capsules can be prepared by filling each standard two-piece hard gelatin capsule with, for example, 100 milligrams of powder of the crystalline form of formula I described herein, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams of magnesium stearate.

[0149] In some embodiments, a mixture of the crystalline form of Formula I described herein and a digestible oil, such as soybean oil, cottonseed oil, or olive oil, can be prepared and injected into gelatin by a positive displacement pump to form a soft gelatin capsule containing 100 milligrams of the active ingredient. The capsule is washed and dried.

[0150] In some embodiments, tablets can be prepared by conventional procedures so that a dosage unit contains, for example, 100 milligrams of the crystalline form of Formula I or a pharma- ceutically acceptable salt thereof, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline cellulose, 11 milligrams of starch, and 98.8 milligrams of lactose. Appropriate coatings can be applied to increase palatability or delay absorption.

[0151] In some embodiments, a parenteral composition suitable for administration by injection can be prepared by stirring 1.5% by weight of the crystalline form of Formula I disclosed herein and / or at least one enantiomer or pharma- ceutically acceptable salt thereof in 10% by volume of propylene glycol. The solution is made up to volume with water for injection and sterilized.

[0152] In some embodiments, an aqueous suspension can be prepared for oral administration. For example, 5 milliliters of each aqueous suspension can be used, containing 100 milligrams of micronized compound or its pharma- ceutically acceptable salt, 100 milligrams of sodium carboxymethylcellulose, 5 milligrams of sodium benzoate, 1.0 gram of sorbitol solution (USP), and 0.025 milliliters of vanillin.

[0153] When the crystalline form of formula I described herein is administered stepwise or in combination with at least one other therapeutic agent, the same dosage form can generally be used.When drugs are administered in physical combination, the dosage type and administration route should be selected according to the compatibility of the drugs to be combined.Therefore, the term co-administration is understood to include administration of at least two drugs simultaneously or sequentially, or administration as a fixed dose combination of at least two active ingredients.

[0154] The crystalline forms of Formula I disclosed herein can be administered as the sole active ingredient or in combination with at least one second active ingredient.

[0155] The crystalline forms of formula I described herein (e.g., type A or type C) can be used as such or in the form of their pharmaceutically acceptable salts, such as hydrochloride, hydrobromide, acetate, sulfate, citrate, carbonate, trifluoroacetate, etc. Salts can be obtained by adding a desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium salt, potassium salt, calcium salt, ammonium salt, organic amino salt, or magnesium salt, etc. Salts can also be obtained by adding a desired acid, either neat or in a suitable inert solvent. Examples of pharma- ceutically acceptable acid addition salts include salts derived from inorganic acids (e.g., hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, etc.), as well as salts derived from relatively non-toxic organic acids (e.g., acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc.).Also included are salts of amino acids such as arginates, and salts of organic acids such as glucuronic acid or galacturonic acid (see, e.g., Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19).

[0156] IV. METHODS OF TREATMENT AND USES Another aspect of the disclosure provides a method of treating a disease or condition comprising administering a therapeutically effective amount of a substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a pharmaceutical composition thereof to a subject in need thereof, wherein the disease or condition is selected from inflammatory diseases, immune diseases (e.g., autoimmune diseases), allergic diseases, graft rejection, necrotic cell diseases, neurodegenerative diseases, CNS diseases, ischemic brain injury, eye diseases, infectious diseases, malignancies, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and viral infections. In some embodiments, the disease or condition is mediated by RIP1 signaling.

[0157] In another aspect, disclosed herein is a substantially pure crystalline form of Formula I (eg, Type A or Type C) or a pharmaceutical composition thereof for use as a medicament.

[0158] In another aspect, disclosed herein is the use of a substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a pharmaceutical composition thereof for the manufacture of a medicament for treating a disease or condition selected from inflammatory diseases, immune diseases (e.g., autoimmune diseases), allergic diseases, graft rejection, necrotic cell diseases, neurodegenerative diseases, CNS diseases, ischemic brain injury, eye diseases, infectious diseases, malignancies, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and viral infections. In some embodiments, the disease or condition is mediated by RIP1 signaling.

[0159] In a further aspect of the disclosure, the substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a pharmaceutical composition thereof is for use in the treatment of a disease or condition selected from an inflammatory disease, an immune disease (e.g., an autoimmune disease), an allergic disease, a graft rejection, a necrotic cell disease, a neurodegenerative disease, a CNS disease, an ischemic brain injury, an eye disease, an infectious disease, a malignancy, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and a viral infection. In some embodiments, the disease or condition is mediated by RIP1 signaling.

[0160] Another aspect of the disclosure provides a method of regulating, e.g., inhibiting, RIP1 signaling in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a pharmaceutical composition thereof.

[0161] In another aspect, disclosed herein is the use of a substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a medicament thereof for modulating, e.g., inhibiting, RIP1 signaling in a subject in need thereof.

[0162] In another embodiment of the disclosure, a substantially pure crystalline form of Formula I (e.g., Type A or Type C) or a pharmaceutical composition thereof is for use in modulating, e.g., inhibiting, RIP1 signaling in a subject in need thereof by contacting the subject with the crystalline form or pharmaceutical composition.

[0163] The compounds of the formulae disclosed herein, the substantially pure crystalline forms of Formula I (e.g., Type A or Type C), or pharmaceutical compositions thereof may be administered once daily, twice daily, or three times daily to treat, for example, a disease or condition selected from inflammatory diseases, immune diseases (e.g., autoimmune diseases), allergic diseases, graft rejection, necrotic cell diseases, neurodegenerative diseases, CNS diseases, ischemic brain injury, eye diseases, infectious diseases, malignancies, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, ALS, Alzheimer's disease, and viral infections. In some embodiments, the disease or condition is mediated by RIP1 signaling.

[0164] The substantially pure crystalline form of formula I (e.g., Type A or Type C) or pharmaceutical composition thereof can be administered in various ways, for example, orally, topically, rectally, parenterally, by inhalation spray, or via an implanted reservoir, although the most suitable route in any given case will depend on the particular host and the nature and severity of the condition for which the active ingredient is administered. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. The compositions disclosed herein are conveniently provided in unit dosage form and can be prepared by any of the methods well known in the art. Parenteral administration can be by continuous infusion over a selected period of time. Other dosage forms contemplated in the present disclosure are as described in WO 2013 / 075083, WO 2013 / 075084, WO 2013 / 078320, WO 2013 / 120104, WO 2014 / 124418, WO 2014 / 151142, and WO 2015 / 023915.

[0165] Contacting is generally accomplished by administering to the subject an effective amount of the crystalline forms of Formula I disclosed herein. Generally, administration is adjusted to achieve a therapeutic dose of about 0.1-50, preferably 0.5-10, more preferably 1-10 mg / kg, although optimal doses vary from compound to compound and are generally determined empirically for each compound.

[0166] The dosage will vary depending on factors such as the recipient's age, health and weight, the extent of the disease, the type of concurrent treatment, if any, the frequency of treatment, and the nature of the effect desired. In general, the daily dose of the active ingredient may vary, for example, from 0.1 to 2000 milligrams per day. For example, 10 to 500 milligrams taken once or multiple times a day may be effective to obtain the desired results.

[0167] In some embodiments, 2 mg to 1500 mg or 5 mg to 1000 mg of a compound of the formulas disclosed herein, a crystalline form of formula I, or a pharmaceutical composition thereof is administered once daily, twice daily, or three times daily. The crystalline form of formula I described herein is administered for morning / midday administration with a nighttime rest period. EXAMPLES

[0168] The following examples are provided to more fully illustrate the present disclosure and are intended to illustrate, but not limit, the present disclosure.

[0169] Example 1: Synthesis of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide [ka] A mixture of 3,5-difluorobenzaldehyde (400 mg, 2.81 mmol) and hydroxylamine hydrochloride (215.15 mg, 3.10 mmol, 1.1 equiv.) was dissolved in a solution of THF / EtOH / H 2 The mixture was stirred in 2 mL of 2H2O, 4 mL / 10 mL / 2 mL) at room temperature for 16 h. The mixture was extracted with EtOAc, washed with water and brine, dried (Na 2 SO 4 Top), concentrated in vacuo to give 3,5-difluorobenzaldehyde oxime as a white solid, which was used in the next step without further purification.

[0170] A mixture of 3,5-difluorobenzaldehyde oxime and 8 M pyridine-borane complex (0.64 mL) in 5 mL of EtOH and 2 mL of THF was kept below 5° C. 10% HCl (6.5 mL) was added dropwise to the mixture. The mixture was then allowed to warm to room temperature over 30 min. The mixture was diluted with Na 2 CO 3 The mixture was neutralized with ethyl acetate, extracted with EtOAc, washed with water and brine, dried (Na 2 SO 4Top), concentrated in vacuo to give the crude product of N-(3,5-difluorobenzyl)hydroxylamine, which was used directly in the next step without purification.

[0171] N-(3,5-difluorobenzyl)hydroxylamine was dissolved in 2 mL of THF / H 2 HO (1:1) and 0.44 mL of saturated NaHCO 3 The solution was cooled to 0° C., 2,2-dimethylbutanoyl chloride (92 mg) was added, and the mixture was stirred at room temperature for 16 h. The mixture was extracted three times with EtOAc, and the combined organic layers were washed with brine, dried (Na 2 SO 4 Top), concentrated in vacuo, and purified by silica gel chromatography to give the title compound (311 mg) in 43% overall yield. 1 H NMR(400MHz,DMSO-d6)δ9.76(s,1H),7.09(td,J=9.4,2.1Hz,1H),6.95-6.86( m,2H),4.66(s,2H),1.64(q,J=7.5Hz,2H),1.13(s,6H),0.72(t,J=7.5Hz,3H). LC-MS(m / z)258.4(M+H + ).

[0172] As disclosed herein, crystalline forms of the title compound were obtained. Certain batches of the title compound, for example, crystalline Type A, were micronized to reduce particle size. The compound was charged to a feeder hopper and then to a jet mill. Milling was started and continued until the desired API particle size (D90), for example, about 30-40 μm or about 15 μm, was obtained. In some batches, the API particle size (D90) was below 20 μm.

[0173] Example 2: Preparation of crystal type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by anti-solvent addition and reverse anti-solvent addition Two anti-solvent addition experiments were performed. Approximately 15 mg of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide ("starting material") (810048-48-B) was dissolved in 0.1-0.2 mL of solvent as shown in Table 4 to obtain a clear solution. The solution was magnetically stirred with the addition of anti-solvent until a precipitate appeared or until the total volume of anti-solvent reached 10 mL. The resulting precipitate was isolated for XRPD analysis. The XRPD pattern is shown in Figure 7 and indicates that only Type A was produced. A DSC curve in Figure 8 showing only a sharp endotherm at 95.1 °C (onset) was observed for the Type A sample (810048-19-B2). [Table 5]

[0174] Twelve more anti-solvent addition experiments were performed. Approximately 15 mg of starting material (817506-01-A) was dissolved in 0.1-0.2 mL of solvent to obtain a clear solution, as shown in Table 5. The solution was magnetically stirred with anti-solvent addition until a precipitate appeared or the total volume of anti-solvent reached 10 mL. The resulting precipitate was isolated for XRPD analysis. The results in Table 5 showed that mixtures of Type A and Type A + Impurity B were produced using different sets of solvents and anti-solvents. [Table 6] * After antisolvent addition and slurrying at 5°C, a clear solution was obtained, which was transferred and evaporated at room temperature.

[0175] Approximately 20 mg of starting material (8152100-01-A) was added to a 20 mL glass vial and dissolved in 0.2-2.0 mL of the corresponding solvent to obtain a clear solution. The solution was magnetically stirred with the addition of anti-solvent until a precipitate appeared or until the total volume of anti-solvent reached 3 mL. The resulting precipitate was isolated for XRPD analysis. If no solid was obtained, slurrying at 5°C / -20°C or evaporation at room temperature was performed. The results in Table 6 showed that Type A was produced. [Table 7] * An oil-like sample was obtained after antisolvent addition and slurrying at 50-5°C temperature cycles (3 cycles), which was transferred and evaporated at room temperature. #: After anti-solvent addition and slurrying at 5°C / -20°C a clear solution was obtained which was decanted and evaporated at room temperature.

[0176] One reverse anti-solvent addition experiment was performed. Approximately 20 mg of starting material (8152100-01-A) was added to a 5 mL glass vial and dissolved in 0.2-2.0 mL of the corresponding solvent to obtain a clear API solution. 5 mL of the anti-solvent from Table 7 was then added to the 20 mL glass vial. The API solution was added to the 20 mL vial containing the anti-solvent under magnetic stirring. The resulting precipitate was isolated for XRPD analysis. If no solid was obtained after addition of the API solution, slurrying at 5 °C / -20 °C or evaporation at room temperature was performed. The results in Table 7 indicated that Type A was produced. [Table 8] * After reverse antisolvent addition and slurrying at 5°C / -20°C a clear solution was obtained so the sample was then transferred to evaporate at room temperature. * : No solid was obtained after addition of antisolvent. A solid was obtained after slurrying at 5°C. ^:After inverse antisolvent addition and slurrying at 5°C / -20°C, a clear solution was obtained, so the sample was transferred and evaporated at room temperature. Compared to the pattern of Type A, some extra weak diffraction peaks were observed, the intensity of which was slightly higher than the background noise. Since only a very limited amount of sample was obtained, no further studies were carried out. ** After reverse antisolvent addition and slurrying for 2 days at 5° C., a clear solution was obtained, so the sample was transferred and allowed to evaporate at room temperature.

[0177] Example 3: Preparation of Crystal Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by Solid Vapor Diffusion Solid vapor diffusion experiments were performed with five different solvents as shown in Table 8. Approximately 12 mg of starting material (810055-01-A) was weighed into a 3 mL vial and placed in a 20 mL vial with 4 mL of volatile solvent. The 20 mL vial was sealed with a cap and kept at RT (room temperature) for 7 days to allow the solvent vapor to interact with the sample. The dissolved sample in the 3 mL vial was removed and evaporated at room temperature to obtain a solid. The solid was tested by XRPD and the results outlined in Table 8 showed that only Type A was obtained. The XRPD and DSC results are shown in Figures 9 and 10. Only a sharp endotherm at about 95°C (onset) was observed in the DSC curve in Figure 10. [Table 9] * After 7 days a clear solution was obtained which was decanted and allowed to evaporate at room temperature.

[0178] Additional solid vapor diffusion experiments were performed using 12 different solvents, as shown in Table 9. Approximately 12 mg of starting material (817506-01-A) was weighed into a 3 mL vial and placed in a 20 mL vial with 4 mL of volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature for 7 days to allow the solvent vapor to interact with the sample. The solids were examined by XRPD and the results, outlined in Table 9, showed that only mixtures of Type A and Type A + Impurity B were obtained with the different solvents used. [Table 10] * After 7 days a clear solution was obtained which was decanted and allowed to evaporate at room temperature.

[0179] Example 4: Preparation of Crystal Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by Slurrying at Room Temperature Slurry conversion experiments were performed in different solvent systems at room temperature. Approximately 20 mg of starting material (810055-01-A) was suspended in 0.2-0.4 mL of the solvent shown in Table 10 in an HPLC vial. After magnetic stirring of the suspension for 4 days at room temperature, the remaining solid was isolated for XRPD analysis. The results outlined in Table 10 indicated that only Type A was produced. The XRPD and DSC results are shown in Figures 11-14. [Table 11] * After slurrying at room temperature for 3 days and then at 5° C. for a further 2 days, a clear solution was obtained which was decanted and allowed to evaporate at room temperature.

[0180] Additional slurry conversion experiments were performed in different solvent systems at room temperature. Approximately 20-40 mg of starting material (817506-01-A) was suspended in 0.2-0.4 mL of the solvent shown in Table 11 in an HPLC vial. After magnetic stirring of the suspension for 4 days at room temperature, the remaining solid was isolated for XRPD analysis. The results outlined in Table 11 showed that Type A, a mixture of Type A + impurity B, and an oil-like sample were produced using the different solvent systems. [Table 12] # After 3 days at room temperature followed by slurrying at 50°C to 5°C (3 cycles), a clear solution was obtained which was transferred and slurried at -20°C to give a solid. & After 3 days at room temperature and 3 cycles of slurrying at 50°C to 5°C, followed by slurrying at -20°C, a clear solution was obtained. 2 Add O or n-heptane and slurry at room temperature for 3 days. * After addition of the anti-solvent n-heptane, a clear solution was obtained, which was decanted and evaporated at room temperature.

[0181] Example 5: Preparation of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by slurrying at 50° C. and −20° C. and by temperature cycling Slurry conversion experiments were performed in different solvent systems at 50° C. Approximately 25 mg of starting material (810055-01-A) was suspended in 0.2-0.4 mL of the solvent shown in Table 12 in an HPLC vial. After magnetic stirring of the suspension at 50° C. for approximately 3 days, the remaining solid was isolated for XRPD analysis. The results outlined in Table 12 indicate that only Type A was produced. Comparative XRPD results are shown in FIG. 15 and FIG. 16. The DSC curve in FIG. 17 shows only a sharp endotherm at approximately 95° C. (onset). [Table 13] * After slurrying at 50° C. for 3 days, a clear solution was obtained, which was transferred and slurried at 5° C. for an additional 2 days. * After slurrying at 50° C. for 3 days and then at 5° C. for 2 days, a clear solution was obtained which was decanted and evaporated at room temperature.

[0182] Additional slurry conversion experiments were also conducted in different solvent systems at 50° C., as shown in Table 13. Approximately 25-50 mg of starting material (817506-01-A) was suspended in 0.4 mL of solvent in an HPLC vial. After magnetically stirring the suspension at 50° C. for approximately 4 days, the remaining solid was isolated for XRPD analysis. The results outlined in Table 13 show that Type A, a mixture of Type A + impurity B, and an amorphous sample were produced using the different solvent systems. [Table 14] * After slurrying at 50° C. for 4 days, a clear solution was obtained, which was transferred and slurried at 50° C. and 5° C. (3 cycles). *After 3 days at room temperature followed by slurrying at 50°C to 5°C (3 cycles), a clear solution was obtained which was transferred and slurried at -20°C to give a solid. & After 3 days at room temperature and 3 cycles of slurrying at 50°C to 5°C, followed by slurrying at -20°C, a clear solution was obtained. 2 Add O or n-heptane and slurry at room temperature for 3 days. ** After addition of the anti-solvent n-heptane, a clear solution was obtained, which was decanted and evaporated at room temperature.

[0183] Approximately 20 mg of starting material (8152100-01-A) was suspended in 0.5 mL of the corresponding solvent in an HPLC vial. The suspension was magnetically stirred (approximately 1000 rpm) at -20°C for approximately 7 days, after which the remaining solid was isolated by centrifugation for XRPD analysis. The results, outlined in Table 14, indicated that Type A was produced. [Table 15] * After slurrying at -20°C, a clear solution was obtained, which was decanted and evaporated at room temperature.

[0184] Approximately 20 mg of starting material (8152100-01-A) was suspended in 0.5 mL of the corresponding solvent in an HPLC vial. The suspension was then heated to 50° C. and equilibrated at 50° C. for 2 hours. The temperature was set to decrease from 50° C. to 5° C. over 450 minutes and hold at 5° C. for 2 hours. The temperature was increased to 50° C. over 30 minutes. The heating and cooling steps were repeated twice and then cooled to 5° C. over 450 minutes. The resulting solid was kept isothermal at 5° C. and then isolated for XRPD analysis. The results outlined in Table 15 showed that Type A was obtained. [Table 16] *After temperature cycling and slurrying at -20°C, a clear solution was obtained, which was decanted and evaporated at room temperature. # After slurrying with temperature cycling, a clear solution was obtained. After slurrying at -20°C, a solid was obtained.

[0185] Example 6: Preparation of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by slow evaporation and fast evaporation at room temperature and 20° C. Slow evaporation experiments were performed under six conditions. Approximately 15 mg of starting material (810055-01-A or 810048-48-B) was dissolved in 2.0 mL of the solvent shown in Table 16 in a 3 mL glass vial. If the material did not completely dissolve, the suspension was filtered using a 0.45 μm PTFE membrane and the filtrate was used in the subsequent step. The visually clear solution was allowed to evaporate at room temperature in a vial sealed with Parafilm® (Pork 5 small hole). The solid was isolated for XRPD analysis and the results outlined in Table 16 indicated that only Type A was observed. The XRPD and DSC results are shown in Figures 18 and 19. [Table 17]

[0186] Additional slow evaporation experiments were performed under 10 conditions. Approximately 15 mg of starting material (817506-01-A) was dissolved in 2.0 mL of the solvent shown in Table 17 in a 3 mL glass vial. If the material did not completely dissolve, the suspension was filtered using a 0.45 μm PTFE membrane and the filtrate was used in the subsequent step. The visually clear solution was allowed to evaporate at room temperature in a vial sealed with Parafilm® (Pork 4 small hole). The solid was isolated for XRPD analysis and the results outlined in Table 17 showed that a mixture of Type A and Type A + Impurity B was observed using the different solvent systems. [Table 18]

[0187] Approximately 20 mg of starting material (8152100-01-A) was dissolved in 2.0 mL of the corresponding solvent in a 3 mL glass vial, and shaking and sonication were performed to dissolve the solid. The sample was filtered using a PTFE membrane (pore size 0.45 μm). The clear solution was allowed to evaporate at room temperature in a vial sealed with Parafilm® (Pork 4 small hole). The solid was isolated for XRPD analysis, and the results summarized in Table 18 showed that Type A was observed. [Table 19] * : After 2 weeks of evaporation at room temperature, no solids were obtained. The sample was transferred and dried under vacuum at room temperature.

[0188] Approximately 20 mg of starting material (8152100-01-A) was dissolved in 0.5-2.0 mL of the corresponding solvent in a 3 mL glass vial, and shaking and sonication were performed to dissolve the solids. The sample was filtered using a PTFE membrane (pore size 0.45 μm). The clear solution was allowed to evaporate at -20°C in a vial sealed with Parafilm® (Pork 4 small hole). The solid was isolated for XRPD analysis, and the results outlined in Table 19 indicated that Type A was observed. [Table 20] * After evaporation at -20°C for 17 days, a clear solution was obtained, which was transferred and evaporated at room temperature.

[0189] Approximately 20 mg of starting material (8152100-01-A) was dissolved in 0.2-2.0 mL of the corresponding solvent in a 3 mL glass vial, and shaking and sonication were performed to dissolve the solids. The sample was filtered using a PTFE membrane (pore size 0.45 μm). The clear solution was evaporated at 80 °C, and the results outlined in Table 20 showed that Type A was observed. [Table 21] * No solids were obtained after evaporation at 80° C. for 3 days. The sample was transferred and dried under vacuum at room temperature.

[0190] Example 7: Preparation of Crystal Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by slow and fast cooling Slow cooling experiments were performed with the three solvent systems. Approximately 20 mg of starting material (810055-01-A or 810048-18-B) was suspended in 0.1-0.2 mL of the solvent shown in Table 21 in a 3 mL glass vial at room temperature. The suspension was then heated to 50° C. and equilibrated for approximately 2 hours, and the suspension was filtered into a new vial using a 0.45 μm PTFE membrane. The solution or filtrate was slow cooled to 5° C. at a rate of 0.1° C. / min. The resulting solid was kept isothermal at 5° C. before being isolated for XRPD analysis. The results outlined in Table 21 indicated that only Type A was obtained. The XRPD and DSC results are shown in FIG. 20 and FIG. 21. [Table 22] * After slow cooling and further standing at -20°C, a clear solution was obtained which was decanted.

[0191] Additional slow cooling experiments were performed in eight solvent systems as shown in Table 22. Approximately 20 mg of starting material (817506-01-A) was suspended in 0.1-2.0 mL of the corresponding solvent in a 3 mL glass vial at room temperature. The suspension was then heated to 50° C., equilibrated for approximately 2 hours, and filtered into a new vial using a 0.45 μm PTFE membrane. The filtrate was slow cooled to 5° C. at a rate of 0.1° C. / min. The resulting solid was kept isothermal at 5° C. and then isolated for XRPD analysis. The results outlined in Table 22 showed that mixtures of Type A and Type A + Impurity B were obtained using the different solvent systems. [Table 23] *After slow cooling, a clear solution was obtained, which was transferred and placed at -20°C. * After slow cooling and further standing at -20°C, a clear solution was obtained. It was transferred to room temperature and allowed to evaporate.

[0192] Approximately 20 mg of starting material (8152100-01-A) was suspended in 0.2-1.0 mL of the corresponding solvent in a 3 mL glass vial. The suspension was then heated to 50°C and equilibrated at 50°C for 2 h before being filtered through a PTFE membrane (pore size 0.45 μm) into a new vial. The filtrate was slow cooled to 5°C at a rate of 0.1°C / min. The resulting solid was kept isothermal at 5°C before being isolated for XRPD analysis. The results in Table 23 indicated that Type A was produced. [Table 24] * After slow cooling to 5° C. and slurrying at −20° C., a clear solution was obtained, so the sample was transferred to evaporate at room temperature. ** After slow cooling to 5° C. and slurrying at −20° C., a clear solution was obtained and allowed to evaporate at room temperature, at which point the sample was transferred and dried under vacuum at room temperature. # After slow cooling to 5° C., a clear solution was obtained, so the sample was transferred and allowed to evaporate at room temperature. ^:After slow cooling to 5°C and slurrying at -20°C, a clear solution was obtained and evaporated at room temperature. The sample was then transferred and dried at room temperature under vacuum. Extra weak diffraction peaks were observed compared to the Type A pattern. As the amount of sample was very limited, the sample was insufficient for XRPD retest or further investigation.

[0193] Approximately 20 mg of starting material (8152100-01-A) was suspended in 0.3-0.5 mL of the corresponding solvent in a 3 mL glass vial. The suspension was then heated to 50 °C and equilibrated at 50 °C for 2 h before being filtered through a PTFE membrane (pore size 0.45 μm) into a new vial. The filtrate was placed at -20 °C for precipitation. The resulting solid was isolated for XRPD analysis. The results in Table 24 indicated that Type A was produced. [Table 25] * : Samples were frozen at -20°C and allowed to clear at room temperature. Samples were transferred and allowed to evaporate at room temperature.

[0194] Example 8: Preparation of Crystal Type A or Crystal Type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by Liquid Vapor Diffusion Four liquid vapor diffusion experiments were performed. Approximately 15 mg of starting material (810055-01-A or 810048-18-B) was dissolved in 0.1-0.3 mL of the solvent shown in Table 25 in a 3 mL glass vial to obtain a clear solution. The 3 mL vial was then placed in a 20 mL vial containing 4 mL of a volatile solvent (shown in the "anti-solvent" column of Table 25). The 20 mL vial was sealed with a cap and kept at room temperature to allow sufficient time for the organic vapor to interact with the solution. The precipitate was isolated for XRPD analysis. After 7-10 days of diffusion, the clear solution was transferred and allowed to evaporate at room temperature and the resulting solid was examined by XRPD. The results are summarized in Table 25 and the XRPD pattern is shown in Figure 22; only Type A was produced. The DSC curve in Figure 23 shows only a sharp endotherm at approximately 95°C (onset). [Table 26] * A clear solution was obtained after diffusion for 7 days, then decanted and allowed to evaporate at room temperature.

[0195] Ten additional liquid vapor diffusion experiments were performed. Approximately 15 mg of starting material (817506-01-A) was dissolved in 0.1-0.5 mL of the solvent shown in Table 26 in a 3 mL glass vial to obtain a clear solution. The 3 mL vial was then placed in a 20 mL vial containing 4 mL of a volatile solvent (shown in the "anti-solvent" column of Table 26). The 20 mL vial was sealed with a cap and kept at room temperature allowing sufficient time for the organic vapor to interact with the solution. The precipitate was isolated for XRPD analysis. After 7 days of diffusion, the clear solution was transferred and allowed to evaporate at room temperature and the resulting solid was examined by XRPD. The results outlined in Table 26 showed that mixtures of Type A, Type C, and Type A + Impurity B were observed using the different solvent systems. [Table 27] * A clear solution was obtained after diffusion for 7 days, then decanted and allowed to evaporate at room temperature.

[0196] Example 9: Preparation of Crystal Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by Grinding Three grinding experiments were performed. Approximately 20 mg of starting material (810055-01-A) was hand ground with a pestle in an agate mortar for about 5 minutes. The solids were confirmed by XRPD (shown in FIG. 24) and the results outlined in Table 27 showed that only Type A was obtained. The DSC curve in FIG. 27 shows only a sharp endotherm at about 95° C. (onset). [Table 28] Four additional grinding experiments were performed. Approximately 25 mg of starting material (817506-01-A) was hand ground with a pestle in an agate mortar for approximately 5 minutes. The solids were confirmed by XRPD and the results, outlined in Table 28, showed that only Type A and a mixture of Type A + impurity B were obtained using different solvent systems. [Table 29]

[0197] Example 10: Preparation of crystalline type A or amorphous forms of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide by polymer-induced, polymer / ionic liquid-induced crystallization, and co-crystal screening methods Polymer induction experiments were performed with two sets of polymer mixtures in six solvents, as shown in Table 29. Approximately 15 mg of starting material (817506-01-A) was dissolved in 2.0 mL of solvent in a 3 mL vial to obtain a clear solution. Approximately 2 mg of polymer mixture was added to a 3 mL glass vial. The solution was allowed to evaporate at room temperature with the vial sealed with Parafilm (poke 3-5 small holes). The solid was isolated for XRPD analysis. The results, outlined in Table 29, showed that Type A and amorphous samples were obtained. [Table 30] * : 0.1 mL of solvent was added and no filtration was performed. Polymer mixture A: polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA), polyvinyl chloride (PVC), polyvinyl acetate (PVAC), hypromellose (HPMC), methyl cellulose (MC) (mass ratio 1:1:1:1:1:1) Polymer mixture B: polycaprolactone (PCL), polyethylene glycol (PEG), poly(methyl methacrylate) (PMMA), sodium alginate (SA), and hydroxyethyl cellulose (HEC) (mass ratio 1:1:1:1:1).

[0198] Eight experiments were set up in different solvents by polymer / ionic liquid induced crystallization. Approximately 20 mg of starting material (815298-01-A) was dissolved in the corresponding solvent in an HPLC vial, and shaking and sonication were performed to dissolve the solid. The sample was filtered using a PTFE membrane (pore size 0.45 μm) to prepare a saturated clear solution. The polymer or ionic liquid was then added to the solution, and the solution was magnetically stirred (approximately 1000 rpm) at room temperature. The resulting solid was isolated for XRPD analysis. The results, outlined in Table 30, showed that only Type A was observed. [Table 31] Polymer mixture A: Polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyvinyl chloride (PVC), polyvinyl acetate (PVAC), hypromellose (HPMC), methylcellulose (MC) (mass ratio 1:1:1:1:1:1) Polymer mixture B: polycaprolactone (PCL), polyethylene glycol (PEG), poly(methyl methacrylate) (PMMA), sodium alginate (SA), and hydroxyethyl cellulose (HEC) (mass ratio 1:1:1:1:1).

[0199] Starting from type A (8152100-01-A), a total of 68 cocrystal screening experiments were set up with 17 coformers and 4 solvent systems via solvent-assisted reactive crystallization. The detailed procedure is as follows: Approximately 20 mg of starting material (8152100-01-A) and the corresponding coformer were mixed in each HPLC vial at the desired molar ratio of 1:1. Then, 0.5 mL of the corresponding solvent (acetone, EtOAc, MTBE) was added to the vial, and all samples were magnetically stirred at room temperature for about 6 days. The solids were isolated for XRPD analysis. The clear solutions were transferred and slurried at 5°C. If a clear solution was still obtained, evaporation at room temperature or anti-solvent addition was performed to induce precipitation. For the experiments with EtOH as the solvent, approximately 20 mg of starting material (8152100-01-A) and equimolar corresponding coformer were transferred to an agate mortar. After hand grinding for approximately 3 minutes, the solids were collected for XRPD analysis. Only Type A, coformer, or a mixture of Type A and coformer was obtained, as outlined in Table 31. [Table 32] [1] The experiment was carried out by grinding. [2] After slurried at room temperature for 6 days, a clear solution was obtained. After slurried at 5° C. for 10 days, a solid was obtained. [3] : A clear solution was obtained after 6 days of slurrying at room temperature and 10 days at 5° C. A solid was obtained after anti-solvent addition (n-heptane). [4] : After slurrying at room temperature for 6 days and at 5° C. for 10 days, a clear solution was obtained. After addition of anti-solvent (n-heptane), no solids were obtained. The solution was decanted and evaporated at room temperature. [5]: Based on the results of the reconstitution and control experiments, the samples with different XRPD patterns were suspected to be new forms of lysine (see Appendix 7.1.2). [6]Based on the results of the reconstitution and control experiments, the samples with different XRPD patterns were suspected to be new forms of arginine (see Appendix 7.1.1). [7] After slurried at room temperature for 6 days, a clear solution was obtained. After slurried at 5° C. for 10 days, a limited amount of solid was obtained. The XRPD pattern of the resulting solid was different from Type A and benzenesulfonic acid. 1 H NMR results showed that several peaks were observed that could not be assigned to the API or benzenesulfonic acid, indicating decomposition of the sample. Reconstitution was performed in MTBE, yielding only Type A.

[0200] Example 11: Preparation of crystal type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide using type A sample Using Type A sample (817506-48-A) as the starting material, five experiments were carried out to prepare Type A at a 200 mg scale. The results are summarized in Table 32. The detailed procedure is as follows:

[0201] 1. Dissolve approximately 200 mg of Type A sample (817506-48-A) in a solvent to obtain a clear solution; 2. While stirring, add anti-solvent dropwise to the solution; 3. The resulting suspension is isolated by centrifugation (10000 rpm, 2 min) for XRPD analysis.

[0202] 4. Dry the solid under vacuum at room temperature for approximately 16 hours.

[0203] As shown in Table 32, the sample (817506-48-A2) was prepared by dissolving Type A in MeOH / H 20, TGA, DSC, and PLM characterization were also performed. As shown in Figure 27, a weight loss of about 0.8% at a maximum of 100°C and a sharp endotherm at 94.8°C (onset) were observed on the TGA / DSC curve. Based on the XRPD (Figures 26 and 29) and DSC results, the batch of sample (817506 48-A2) is pure Type A. The PLM image in Figure 28 showed that Type A (817506-48-A2) contains blade-like crystals. [Table 33] * An oily sample was obtained and converted to Type A after slurrying.

[0204] Example 12: Preparation of crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide using type A + impurity B as starting material To investigate whether pure type A could be obtained from a mixture of type A + impurity B, the A+B sample (817506-38-A) was used as the starting material and purified using MeOH / H 2 Another batch of experiments was carried out in the O system. The detailed procedure is as follows:

[0205] 1. Weigh 100.4 mg of A+B sample (817506-38-A, Lot Number: SN-023-91-17) into a 5 mL glass vial. Dissolve the solid in 0.5 mL of MeOH to obtain a clear solution.

[0206] 2. While stirring, add 0.25 mL of anti-solvent H 2 Add O dropwise to the clear solution. A suspension was obtained.

[0207] 3. Isolate the solids by centrifugation (10000 rpm, 2 min) for XRPD analysis.

[0208] 4. Dry the solid under vacuum at room temperature for approximately 16 hours. A total of 71.3 mg of solid was obtained.

[0209] The XRPD pattern in Figure 30 showed that the obtained solid (810048-03-A1) was Type A with no diffraction peaks of impurity B. The DSC curve in Figure 31 showed only a sharp endotherm at 94.3°C (onset). Based on the data, pure Type A was obtained.

[0210] Example 13: Evaluation of Type A--Equilibrium Solubility in Water H 2 The equilibrium solubility of Type A in 20 was measured at room temperature. Specifically, 10.0 mg of Type A sample (817506-48-A2) was dissolved in 1 mL of 200 mL of 200 mL of 1 ... 2 The suspension was suspended (approximately 1000 rpm) in 100 mL of 1000 mL at room temperature. After 24 hours, the suspension was centrifuged (10000 rpm, 5 min, room temperature) and subsequently filtered (0.45 μm PTFE membrane). The supernatant (the first few drops were discarded) was analyzed for HPLC solubility and pH, and the remaining solid was used for XRPD analysis. The results are summarized in Table 33. The measured solubility of Type A was 0.15 mg / mL. As shown in FIG. 32, H 2 No morphological changes were observed for Type A after suspension in O at room temperature for approximately 24 hours. [Table 34]

[0211] Example 14: Evaluation of Type A--Solid State Stability To evaluate the solid state stability, Type A sample (817506-48-A2) was stored at 80° C. for 1 day, and at 25° C. / 60% RH and 40° C. / 75% RH for 1 week each. All samples were characterized using XRPD, DSC, and HPLC, and the results are summarized in Table 34. No morphological changes or loss of purity was observed for Type A sample (817506-48-A2) under all conditions, indicating that Type A has good solid state stability. The HPLC chromatogram overlay is shown in FIG. 33. The XRPD results are shown in FIG. 34. The DSC characterization results are shown in FIG. 35-FIG. 37. Only one endotherm was observed at about 95° C., indicating that the solid was pure Type A under all conditions. [Table 35]

[0212] Example 15: Evaluation of Type A--Photostability The photostability of Type A (817506-48-A2) was verified in accordance with ICH guidelines under white light (1,200,000 Lux.hrs) and ultraviolet light (200W·hrs / m 2 ) conditions. The results are summarized in Table 35. As shown in the HPLC chromatograms in Figures 38 and 39, no purity loss was observed for Type A after the photostability evaluation. The XRPD results are shown in Figures 40 and 41. Only an endotherm at about 95°C was observed on the DSC curve (Figures 42 and 43). Combined with the XRPD results, no morphological changes were detected after the photostability evaluation under both white light and UV light, indicating that Type A has good photostability. [Table 36] * : Samples in foil-covered vials under both conditions are run as control tests.

[0213] Example 16: Evaluation of Type A - Hygroscopicity To investigate the solid morphology stability as a function of humidity, DVS isotherm plots of Type A (817506-48-A2) were collected at 25° C. from 0 to 95% RH. The DVS plot of Type A in FIG. 44 showed a water uptake of 0.10% at 25° C. / 80% RH, indicating that Type A is non-hygroscopic. As shown in FIG. 45, no difference in the XRPD patterns was observed for Type A before and after the DVS test, indicating no morphology change. The DSC curve in FIG. 46 showed only a melting endotherm for Type A at 95.0° C. (onset) after the DVS test.

[0214] Example 17: Preparation of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide type C N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide type C (817506-10-A10) was 2 The XRPD pattern was obtained via liquid vapor diffusion of a DMSO solution of the starting material (817506-01-A) in an O atmosphere. As shown in Figure 47, the XRPD pattern of Type C is similar to Type A, except for some peak shifts (shown by dashed lines). After drying in vacuum at 50 °C, Type C was converted to Type A. The XRPD comparison results are shown in Figure 48.

[0215] Example 18: Solubility measurement of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide The approximate solubility of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide (817506-01-A) was measured at room temperature in 20 solvent systems. Approximately 2 mg of sample was added to a 3 mL glass vial. The solvents shown in Table 36 were then added stepwise to the vial (50 / 50 / 200 / 700 / 1000 μL) until the solid was visibly dissolved or a total volume of 2 mL was reached. The solubility results outlined in Table 36 were used to guide solvent selection in preparing the solid forms disclosed herein. [Table 37]

[0216] Example 19: Characterization of Type A Form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide Three batches of the crystalline Type A form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide were characterized using XRPD, TGA, and DSC. The results are shown in Table 37 and Figures 49-53. One batch of material was also characterized by NMR, as shown in Figure 54. [Table 38]

[0217] Example 20: Characterization of Type A Form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide Containing Impurity B Four batches of material containing the crystalline Type A form of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide and impurity B were characterized using XRPD and DSC. The results are shown in Table 38 and Figures 55 and 56. [Table 39] * : Peak endotherm.

[0218] Example 21: Apparatus and methods XRPD An X-ray powder diffractometer / Bruker D8 Advance (ADS-XRPD-001) instrument was used for the XRPD analysis shown in Figures 5A and 6A. The XRPD parameters used are listed in Table 39 parameters for these XRPD studies. [Table 40] * NOTE: The detector SSD160 belongs to LynxEye.

[0219] An X'pert 3 X-ray powder diffractometer was used for the XRPD analyses described in Examples 2-20 and shown in Figures other than Figures 5A and 6A. Test samples were spread in the center of a Si holder with zero background. The XRPD parameters used are listed in Table 40 parameters for these XRPD studies. [Table 41]

[0220] TGA and DSC TGA data was collected using a TA Instruments TA Q5000 and Discovery TGA 5500 TGA, and DSC was performed using a TA Instruments TA Q2000 and Discovery DSC 2500 DSC. The detailed parameters used are listed in Table 41. [Table 42]

[0221] HPLC An Agilent 1290 UPLC equipped with a DAD detector and an Agilent 1260 HPLC equipped with a VWD detector were used. Detailed chromatographic conditions for purity and solubility analysis are listed in Tables 42 and 43. [Table 43] [Table 44]

[0222] DVS DVS was measured by SMS (Surface Measurement Systems) DVS Intrinsic. Relative humidity at 25 °C was measured using LiCl, Mg(NO 3 ) 2 The DVS test parameters are listed in Table 44. [Table 45]

[0223] PLM PLM images were taken with a ZEISS Scope. A1 microscope.

[0224] 1 H NMR 1H (proton) solution NMR was collected on a Bruker 400M NMR spectrometer using DMSO-d6 as the solvent.

[0225] All publications, including but not limited to disclosures and disclosed applications, cited herein are incorporated by reference as if fully set forth. In the event that any particular content of a publication cited herein contradicts or is inconsistent with the present disclosure, the present disclosure shall control.

[0226] Those skilled in the art will readily recognize that various changes, modifications, and variations can be made therein without departing from the spirit and scope of the disclosure, as defined in the following claims.

Claims

1. Crystalline compound N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide.

2. The crystalline compound of claim 1, wherein the crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide is a crystalline compound of claim 1.

3. 3. The crystalline type A of claim 2, wherein the crystalline type A is substantially pure.

4. 3. The crystalline Type A of claim 2, wherein the crystalline Type A has an XRPD pattern substantially the same as one of the XRPD patterns displayed for crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide shown in Figures 1, 5A, 6A, 7, 9, 11, 12, 13, 15, 16, 18, 20, 22, 24, 26, 29, 30, 32, 34, 40, 41, 45, 47, 48, 49, 50, 52, and 55.

5. 3. Crystalline Type A according to claim 2, wherein the Crystalline Type A has an XRPD pattern comprising peaks at 9.15, 17.15, 18.22, 21.8, and 27.40 degrees 2θ (two theta) (±0.2 degrees 2θ).

6. 3. Crystalline Type A according to claim 2, wherein the crystal type A has an XRPD pattern including a peak at 13.96 degrees two-theta (2-theta) (±0.2 degrees θ2) and one of the following peaks at 9.15, 14.15, 17.15, 18.22, and 26.31 degrees two-theta (±0.2 degrees 2θ).

7. 3. Crystalline Type A according to claim 2, wherein the crystal type A has an XRPD pattern including a peak at 13.96 degrees two-theta (2-theta) (±0.2 degrees θ2) and two of the following peaks at 9.15, 14.15, 17.15, 18.22, and 26.31 degrees two-theta (±0.2 degrees 2θ).

8. 3. Crystalline Type A according to claim 2, wherein the crystalline Type A has an XRPD pattern including any three of peaks at degrees 2θ (±0.2 degrees 2θ) selected from the group consisting of 9.15, 13.96, 14.15, 17.15, 18.22, and 26.

31.

9. 3. Crystalline Type A according to claim 2, wherein the Crystalline Type A has an XRPD pattern comprising peaks at 9.15, 13.96, 14.15, 17.15, 18.22, and 26.31 degrees 2θ (±0.2 degrees 2θ).

10. 3. Crystalline Type A according to claim 2, wherein the crystal type A has an XRPD pattern including a peak at 18.20 degrees two-theta (2-theta) (±0.2 degrees θ2) and one of the following peaks at 9.10, 17.10, 21.7, and 27.40 degrees two-theta (±0.2 degrees 2θ).

11. 3. Crystalline Type A according to claim 2, wherein the crystal type A has an XRPD pattern including a peak at 18.20 degrees two-theta (2-theta) (±0.2 degrees θ2) and two of the following peaks at 9.10, 17.10, 21.7, and 27.40 degrees two-theta (±0.2 degrees 2θ).

12. 3. Crystalline Type A according to claim 2, wherein the crystalline Type A has an XRPD pattern including any three of peaks at degrees 2θ (±0.2 degrees 2θ) selected from the group consisting of 9.10, 17.10, 18.20, 21.7, and 27.

40.

13. 3. Crystalline Type A according to claim 2, wherein the Crystalline Type A has an XRPD pattern comprising peaks at 9.10, 17.10, 18.20, 21.7, and 27.40 degrees 2θ (±0.2 degrees 2θ).

14. 3. The crystalline type A of claim 2, wherein the crystalline type A is anhydrous.

15. 15. The crystalline type A of any one of claims 2 to 14, wherein the crystalline type A has, prior to decomposition of the compound, a differential scanning calorimetry thermogram substantially the same as one of the DSC thermograms shown in Figure 2, 5B (prior to decomposition of the compound), 6B (prior to decomposition of the compound), 8, 10, 14, 17, 19, 21, 23, 25, 27, 31, 35, 36, 37, 42, 43, 46, 51, 53, and 56.

16. 15. The crystalline type A according to any one of claims 2 to 14, wherein the crystalline type A has an onset melting temperature of about 94°C to about 96°C.

17. Crystalline type A according to any one of claims 2 to 14, having an onset melting temperature of about 94.5°C to about 95.5°C.

18. Crystalline type A according to any one of claims 2 to 14, having an onset melting temperature of about 95°C.

19. The crystalline type A according to any one of claims 2 to 14, wherein the crystalline type A has a thermogravimetric analysis graph substantially the same as one of the TGA graphs shown in Figures 3, 27, 51, and 53.

20. 3. A method for preparing crystalline type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide according to claim 2, comprising the steps of: (a) dissolving N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide in a first solvent to obtain a solution; (b) adding a second solvent to the solution while stirring the solution to obtain a suspension; (c) isolating the solids from the suspension; (d) drying the solid to obtain crystalline type A; A method comprising:

21. 21. The method of claim 20, wherein the first solvent is a solvent in which N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide has a solubility of greater than 10 mg / mL, and the second solvent is a solvent in which N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide has a solubility of less than 10 mg / mL.

22. The first solvent may be isoamyl alcohol, ethyl lactate, MEK, anisole, n-butanol / n-BuOH, ethyl formate, 2,2,2-trifluoroethanol, toluene, pyridine, isobutanol, chlorobenzene, CPME, m-xylene, n-butyl acetate, cumene, NMP, MTBE, 2-MeTHF, EtOAc, acetone, THF, DMAc, IPA, EtOH, DCM, MeOH, IPA, MIBK, IPAc, THF, 1,4-dioxane, DCM, CHCl ３ , toluene, DMSO, DMAc, NMP, and ACN, and the second solvent is selected from the group consisting of n-hexane, water, 2-MeTHF, MTBE, cyclohexane, and n-heptane.

23. 3. A method for making crystalline Type A of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide according to claim 2, wherein the method is selected from the group consisting of anti-solvent addition, reverse anti-solvent addition, solid vapor diffusion, slurrying at room temperature, slurrying at about 50°C, slurrying at about -20°C, slurrying with temperature cycling, slow evaporation at room temperature, slow evaporation at about 20°C, fast evaporation, slow cooling, fast cooling, liquid vapor diffusion, grinding, polymer-induced method, polymer / ionic liquid-induced crystallization, and co-crystal screening.

24. The crystalline compound of claim 1, wherein the crystalline type C of N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide is a crystalline compound of claim 1.

25. 25. The crystalline type C of claim 24, wherein the crystalline type C is substantially pure.

26. 25. The crystalline Type C of claim 24, wherein the crystalline Type C has an XRPD pattern substantially the same as the XRPD pattern displayed for Type C shown in Figure 47.

27. 25. Crystalline Type C according to claim 24, wherein the Crystalline Type C has an XRPD pattern comprising peaks at 9.08, 18.18, 21.78, and 27.39 degrees two-theta (2θ) (±0.2 degrees two-theta).

28. 25. Crystalline Type C according to claim 24, wherein the Crystalline Type C has an XRPD pattern comprising peaks at 9.08, 13.45, 14.28, 18.18, 21.78, and 27.39 degrees two-theta (2θ) (±0.2 degrees two-theta).

29. A composition comprising the crystalline compound N-(3,5-difluorobenzyl)-N-hydroxy-2,2-dimethylbutanamide according to any one of claims 1 to 14 and claims 24 to 28, and a pharmaceutically acceptable carrier.

30. 29. The crystalline compound of any one of claims 1 to 14 and 24 to 28, or a pharmaceutical composition comprising said crystalline compound, for use in the treatment of a disease or condition, wherein the disease or condition is selected from inflammatory diseases, immune diseases, allergic diseases, transplant rejection, necrotic cell diseases, neurodegenerative diseases, central nervous system (CNS) diseases, ischemic brain injury, eye diseases, infectious diseases, malignant tumors, ulcerative colitis, Crohn's disease, psoriasis, rheumatoid arthritis, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, and viral infections.

31. 31. The crystalline compound or pharmaceutical composition of claim 30, wherein the disease or condition is mediated by receptor-interacting protein 1 (RIP1) signaling.

32. 29. A method of inhibiting receptor-interacting protein 1 (RIP1), comprising contacting the RIP1 protein or a fragment thereof with a crystalline compound according to any one of claims 1 to 14 and 24 to 28, or a pharmaceutical composition comprising the crystalline compound.

Images