Amorphous and crystalline forms of MCI-1 antagonists

Abstract

Disclosed herein is (1S,3'R,6'R,7'S,8'E,11'S,12'R)-6-chloro-7'-methoxy-11',12'-dimethyl-3,4-dihydro-2H,15'H-spiro[naphthalene-1,22'-[20]oxa[13]thia[1,14]diazatetracyclo[14.7.2.0 3,6 .0 19,24 [0013] The present invention relates to crystalline and amorphous forms of pentacosa[8,16,18,24]tetraen]-15'-one 13',13'-dioxide (AMG176), as well as salts and hydrates thereof. Also disclosed are methods of making the crystalline and amorphous forms, and methods of treating diseases and disorders with the crystalline and amorphous forms. TIFF2025511331000013.tif26170

Description

[Technical field]

[0001] The present disclosure relates to the use of (1S,3'R,6'R,7'S,8'E,11'S,12'R)-6-chloro-7'-methoxy-11',12'-dimethyl-3,4-dihydro-2H,15'H-spiro[naphthalene-1,22'-

[20] oxa

[13] thia[1,14]diazatetracyclo[14.7.2.0])-1,2'-diamino-2,3'-diphenylphosphine oxide (2H,15'H-DMPO) that functions as an inhibitor of myeloid cell leukemia 1 protein (Mcl-1). 3,6 .0 19,24 ]Pentacosa[8,16,18,24]tetraen]-15'-one 13',13'-dioxide (AMG176) in crystalline and amorphous forms, its hydrates, and salts. [Background technology]

[0002] 2. Description of Related Art The compound (1S,3'R,6'R,7'S,8'E,11'S,12'R)-6-chloro-7'-methoxy-11',12'-dimethyl-3,4-dihydro-2H,15'H-spiro[naphthalene-1,22'

[20] oxa

[13] thia[1,14]diazatetracyclo[14.7.2.03,6.019,24]pentacosa[8,16,18,24]tetraene]-15'-one 13',13'-dioxide (AMG176) is useful as an inhibitor of myeloid cell leukemia 1 (Mcl-1): [ka]

[0003] One common characteristic of human cancers is overexpression of Mcl-1, which prevents cancer cells from undergoing programmed cell death (apoptosis), allowing them to survive despite extensive genetic damage.

[0004] Mcl-1 is a member of the Bcl-2 family of proteins. The Bcl-2 family includes pro-apoptotic members (such as BAX and BAK) that form homo-oligomers in the outer mitochondrial membrane shortly after activation, leading to pore formation and escape of mitochondrial contents, a step that triggers apoptosis. Anti-apoptotic members of the Bcl-2 family (such as Bcl-2, Bcl-XL, and Mcl-1) block the activity of BAX and BAK. Other proteins (such as BID, BIM, BIK, and BAD) exhibit additional regulatory functions. Studies have shown that Mcl-1 inhibitors may be useful in the treatment of cancer. MCl-1 is overexpressed in many cancers.

[0005] US Patent No. 9,562,061, the entirety of which is incorporated herein by reference, discloses AMG176 as an Mcl-1 inhibitor and provides a method for its preparation. However, alternative forms of AMG176 with improved properties are desirable, particularly for the clinical use of AMG176. Summary of the Invention [Means for solving the problem]

[0006] Crystalline and amorphous forms of AMG176 are provided herein, and AMG176 has the following structure: [ka]

[0007] Also, the solids at 12.39, 19.40, 20.46, 27.24, 28.07, 30.54, 33.09, 33.80, 37.18, 41.80, 42.98, 54.86, 58.69, 60.01, 63.11, 80.06, 85.77, 117.20, 119.77, 120.86, 127.04, 129.01, 129.71, 131.30, 132.24, 133.58, 139.16, 140.11, 140.69, 152.08, and 169.82 ± 0.5 ppm. 13Also provided is a crystalline form of AMG176 characterized by C NMR peaks ("AMG176 Form 1").

[0008] Also provided is a crystalline form of AMG176 characterized by an XRPD pattern peaks at 13.0, 16.9, and 17.3±0.2 degrees 2θ using Cu Kα radiation ("AMG176 Form 2").

[0009] Also provided is an amorphous form of AMG176 ("AMG176 amorphous") having an XRPD pattern substantially as shown in Figure 14.

[0010] Also provided is a crystalline form of AMG176 as a calcium salt hydrate characterized by an XRPD pattern peaks at 6.8, 7.8, and 15.5±0.2 degrees 2θ using Cu Kα radiation ("AMG176 calcium hydrate Form 1").

[0011] Also provided is a crystalline form of AMG176 as a calcium salt hydrate characterized by an XRPD pattern peaks at 6.2, 20.2, and 24.4±0.2 degrees 2θ using Cu Kα radiation ("AMG176 calcium hydrate Form 2").

[0012] Also provided is a crystalline form of AMG176 as a calcium salt hydrate characterized by an XRPD pattern peaks at 6.2, 6.7, and 8.2±0.2 degrees 2θ using Cu Kα radiation ("AMG176 calcium hydrate Form 3").

[0013] Also provided is an amorphous form of AMG176 as a calcium salt hydrate ("AMG176 amorphous calcium hydrate") having an XRPD pattern substantially as shown in Figure 28.

[0014] Also provided are pharmaceutical formulations comprising a crystalline or amorphous form of AMG176 described herein and a pharma- ceutically acceptable excipient.

[0015] Also provided is a method of treating a subject suffering from cancer comprising administering to the subject a therapeutically effective amount of a crystalline or amorphous form of AMG176 described herein, or a pharmaceutical formulation comprising a crystalline or amorphous form of AMG176 described herein and a pharma- ceutical acceptable excipient. [Brief description of the drawings]

[0016] [Figure 1] 1 shows the solid state 13C NMR of crystalline Form 1 of AMG176. [Diagram 2] 1 shows the X-ray powder diffraction ("XRPD") pattern of crystalline Form 1 of AMG176. [Diagram 3] 1 shows a differential scanning calorimetry ("DSC") thermograph of crystalline Form 1 of AMG176. [Figure 4] 1 shows a thermogravimetric analysis ("TGA") trace of crystalline Form 1 of AMG176. [Diagram 5] 1 shows the moisture sorption profile (DVS) of crystalline Form 1 of AMG176. [Figure 6] 1 shows the single crystal X-ray crystal structure of the unit cell of crystalline Form 1 of AMG176. [Figure 7-1] 1 shows an overlay of X-ray powder diffraction ("XRPD") patterns of crystalline Form 1 of AMG 176 after solid state physical stability evaluation. No form transformation was observed after 8 weeks under stress conditions. [Figure 7-2] 1 shows an overlay of X-ray powder diffraction ("XRPD") patterns of crystalline Form 1 of AMG 176 after solid state physical stability evaluation. No form transformation was observed after 8 weeks under stress conditions. [Figure 8] 1 shows the X-ray powder diffraction ("XRPD") pattern of crystalline Form 2 of AMG176. [Figure 9] 1 shows a differential scanning calorimetry ("DSC") thermograph of crystalline Form 2 of AMG176, showing an onset and melting peak temperatures of approximately 233°C and 238°C, respectively. [Figure 10]1 shows a differential scanning calorimetry ("DSC") thermograph and thermogravimetric analysis ("TGA") trace overlay of crystalline 2-Me THF solvate Form 2 of AMG176, showing a DSC onset temperature of 176°C and a TGA weight loss of 12.8% from 140 to 215°C. [Figure 11] 1 shows a differential scanning calorimetry ("DSC") thermograph and thermogravimetric analysis ("TGA") trace overlay of crystalline THF / water solvate Form 2 of AMG176, showing a DSC onset temperature of 177°C and a TGA weight loss of 8.9% from 140-200°C. [Figure 12] 1 shows a differential scanning calorimetry ("DSC") thermograph and thermogravimetric analysis ("TGA") trace overlay of crystalline MTBE solvate Form 2 of AMG176, showing a DSC onset temperature of 163°C and a TGA weight loss of 12.7% from 140-190°C. [Figure 13] 1 shows a differential scanning calorimetry ("DSC") thermograph and thermogravimetric analysis ("TGA") trace overlay of crystalline 1,4-dioxane / water solvate Form 2 of AMG176, showing a DSC onset temperature of 170°C and a TGA weight loss of 10.1% from 120-180°C. [Figure 14] 1 shows the X-ray powder diffraction ("XRPD") pattern of an amorphous form of AMG176. [Figure 15] 1 shows a differential scanning calorimetry ("DSC") thermograph of the amorphous form of AMG176, showing a Tm of approximately 163°C. [Figure 16] 1 shows a thermogravimetric analysis ("TGA") trace of the amorphous form of AMG176, showing a 5.4% weight loss by 200°C and an 11.6% weight loss by 275°C. [Figure 17] 1 shows a thermogravimetric analysis ("TGA") trace of the amorphous form of AMG176, showing a weight loss of 3.8% by 180° C. [Figure 18]Figure 1 shows the moisture sorption profile (DVS) of the amorphous form of AMG176, showing a weight gain of approximately 0.95 wt% water vapor from 5% to 95% relative humidity. More weight was lost (1.59%) than gained upon adsorption, likely due to loss of residual DCM and / or water from the starting material. XRPD of the material after DVS was consistent with amorphous material. [Figure 19] 1 shows an overlay of XRPD analysis of an amorphous AMG176 sample after solid state physical stability evaluation: no crystallization was observed after 8 weeks under stress conditions. [Figure 20] 1 shows the X-ray powder diffraction ("XRPD") pattern of the crystalline calcium salt hydrate Form 1 of AMG176. [Figure 21] FIG. 1 shows overlaid differential scanning calorimetry ("DSC") thermograph and thermogravimetric analysis ("TGA") traces of the crystalline calcium salt hydrate Form 1 of AMG176, showing melting at 301°C, recrystallization, and another melting at 322°C, followed by decomposition, and a TGA weight loss of approximately 5.5%. [Figure 22] 1 shows the X-ray powder diffraction ("XRPD") pattern of the crystalline calcium salt hydrate Form 2 of AMG176. [Diagram 23] FIG. 1 shows the moisture sorption profile (DVS) of the crystalline calcium salt hydrate Form 2 of AMG176, showing a weight gain of approximately 5.2% at 70% relative humidity and 23% at 95% relative humidity, likely due to the presence of residual NaCl. [Figure 24] 1 shows the X-ray powder diffraction ("XRPD") pattern of the crystalline calcium salt hydrate Form 3 of AMG176. [Diagram 25] 1 shows a differential scanning calorimetry ("DSC") thermograph of the crystalline calcium salt hydrate Form 3 of AMG176, showing a Tm of 314°C. [Figure 26] 1 shows a thermogravimetric analysis ("TGA") trace of the crystalline calcium salt hydrate Form 3 of AMG176, showing a weight loss of 3.5%. [Figure 27] FIG. 1 shows the moisture sorption profile (DVS) of the crystalline calcium salt hydrate Form 3 of AMG176, showing a weight gain of approximately 7.4% water at 95% relative humidity. [Figure 28] 1 shows the X-ray powder diffraction ("XRPD") pattern of the amorphous calcium salt form of AMG176. [Figure 29] 1 shows an overlay of differential scanning calorimetry ("DSC") thermographs of calcium salt forms of AMG176 (top to bottom: crystalline form 2, amorphous form, and crystalline form 1), showing Tms of 323°C, 295°C, and 322°C, respectively. [Diagram 30] 1 shows an overlay of thermogravimetric analysis ("TGA") traces of calcium salt forms of AMG176 (top to bottom: amorphous form, crystalline form 2, and crystalline form 1) showing approximately 2.0% weight loss, approximately 3.1% weight loss, and approximately 5.5% weight loss, respectively. [Diagram 31] FIG. 1 shows the moisture sorption profile (DVS) of the amorphous calcium salt form of AMG176, showing weight gain from approximately 9% to 95% relative humidity. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] AMG176 is a small molecule that targets the Mcl-1 pathway for the treatment of hematological malignancies. High-throughput (HTS) and manual screening experiments conducted to date have failed to isolate and / or scale-up a salt or crystalline form suitable for development. Polymorph screening against the free acid disclosed herein has identified the polymorphs disclosed herein, such as the crystalline free acid, AMG176 Form 1, as suitable forms for development to support intravenous (IV) drug development. The AMG176 polymorphs disclosed herein can also be suitable for oral delivery as amorphous solid dispersions, if desired.

[0018] The present invention relates to ((1S,3'R,6'R,7'S,8'E,11'S,12'R)-6-chloro-7'-methoxy-11',12'-dimethyl-3,4-dihydro-2H,15'H-spiro[naphthalene-1,22'-

[20] -oxa

[13] thia[1,14]diazatetracyclo[14.7.2.0 3,6 .0 19,24]Pentacosa[8,16,18,24]tetraen]-15'-one 13',13'-dioxide (AMG176), its hydrates and salts are disclosed: [ka] AMG176 crystalline Form 1 is an anhydrous / nonsolvated form of AMG176 that may be thermodynamically stable between 2 and 79 °C. AMG176 crystalline Form 1 may be advantageous over other solvated forms due to improved properties for formulation.

[0019] Also provided herein are pharmaceutical formulations of crystalline and amorphous forms of AMG176, and methods of treating a subject suffering from cancer comprising administering to the subject a therapeutically effective amount of a pharmaceutical formulation of the crystalline or amorphous form disclosed herein.

[0020] US Pat. No. 9,562,061, the entirety of which is incorporated herein by reference, discloses synthetic procedures for synthesizing Mcl-1 inhibitors, such as AMD176.

[0021] Further provided herein are crystalline hydrate forms of AMG176, pharmaceutical formulations thereof, and methods of treating a subject suffering from cancer comprising administering to the subject a therapeutically effective amount of a pharmaceutical formulation of the crystalline hydrate form disclosed herein.

[0022] The compounds disclosed herein may be identified herein either by their chemical structure and / or chemical name. In the event that the chemical structure and the chemical name conflict, the chemical structure is determinative of the compound's identity.

[0023] When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formula, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included.

[0024] As used herein, dashed and bold bonds (i.e., [ka] Chemical structures containing one or more stereocenters, as depicted in the drawings, are meant to depict the absolute stereochemistry of the stereocenters present in the chemical structure. As used herein, bonds represented by simple line symbols do not depict stereogenic preference. Unless specifically indicated to the contrary, chemical structures containing one or more stereocenters illustrated herein without depicting absolute or relative stereochemistry encompass all possible stereoisomeric forms (e.g., diastereomers, enantiomers) of the compound and mixtures thereof. Structures with a single bold or dashed line and at least one additional simple line encompass a single enantiomeric series of all possible diastereomers.

[0025] The term "about" is intended to account for variations due to experimental error. All measurements reported herein are understood to be modified by the term "about," unless expressly stated otherwise, regardless of whether this term is expressly used. As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0026] "Treatment" or "treating" refers to any treatment of a disease in a patient, including a) preventing the disease, i.e., not allowing clinical symptoms of the disease to develop; b) inhibiting the disease; c) slowing or arresting the progression of clinical symptoms; and / or d) relieving the disease, i.e., causing regression of clinical symptoms. Treatment of diseases and disorders, as used herein, is also intended to include prophylactic administration of the pharmaceutical formulations described herein to a subject (i.e., an animal, preferably a mammal, most preferably a human) believed to be in need of treatment, such as, for example, cancer.

[0027] The term "therapeutically effective amount" means an amount effective when administered to a human or non-human patient to treat a disease, for example, a therapeutically effective amount can be an amount sufficient to treat a disease or disorder responsive to inhibition of McI-1. A therapeutically effective amount can be ascertained experimentally, for example, by assaying blood levels of a chemical, or theoretically, or by calculating bioavailability.

[0028] The term "solvate" refers to a chemical entity formed by the interaction of a solvate with a compound. Crystalline solvates of AMG176 are specifically contemplated for use in the formulations herein. Solvents that can form crystalline solvate forms of AMG176 include, but are not limited to, 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), methyl tert-butyl ether (MTBE), 1,4-dioxane, water, and combinations thereof. In some cases, the solvate has 0.5 to 2 solvent molecules per AMG176 molecule.

[0029] The term "hydrate" is a special type of solvate in which the solvent is water. Hydrates, as used herein, have variable amounts of water, including, for example, hemihydrates, monohydrates, dihydrates, trihydrates, etc. Crystalline hydrates of AMG176 are specifically contemplated for use in the formulations disclosed herein. In some cases, the hydrates have 0.5 to 2 water molecules per AMG176 molecule.

[0030] The term "polymorph" as used herein includes all crystalline and amorphous forms of a compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrous), and conformational polymorphs, and mixtures thereof, unless a specific crystalline or amorphous form is referred to. In some embodiments, the present disclosure provides crystalline forms of AMG176, such as crystalline polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrous), and conformational polymorphs, and mixtures thereof, unless a specific crystalline form is referred to.

[0031] crystalline form Crystalline Form 1: AMG176 crystalline Form 1 has peaks at 12.39, 19.40, 20.46, 27.24, 28.07, 30.54, 33.09, 33.80, 37.18, 41.80, 42.98, 54.86, 58.69, 60.01, 63.11, 80.06, 85.77, 117.20, 119.77, 120.86, 127.04, 129.01, 129.71, 131.30, 132.24, 133.58, 139.16, 140.11, 140.69, 152.08, and 169.82 ± 0.5 ppm. 13 In some embodiments, AMG176 Form 1 can be characterized by C NMR. 13 C NMR substantially having a C NMR spectrum, where "substantially" means that the reported peaks may vary by ±0.5 ppm.

[0032] AMG 176 Form 1 can be characterized by X-ray powder diffraction obtained as described in the Examples, having peaks at 14.2, 18.0, and 18.6±0.2 degrees 2θ using Cu Kα radiation, and optionally further characterized by additional peaks at 12.6, 17.0, 22.7, and 26.6±0.2 degrees 2θ using Cu Kα radiation, and / or additional peaks at 5.6, 12.3, 12.8, 14.5, 16.7, 24.4, 25.2, 27.1, 28.3, and 28.9±0.2 degrees 2θ using Cu Kα radiation. In some embodiments, AMG 176 Form 1 has an X-ray powder diffraction pattern substantially as shown in Figure 2, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum depend on a number of factors such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0033] A differential scanning calorimetry (DSC) thermogram was obtained for AMG176 Form 1 as described in the Examples. The DSC curve shows an endothermic transition at 236° C.±3° C. Thus, in some embodiments, the amorphous form of AMG176, Form 1, may be characterized by a DSC thermogram having a transition endotherm with an onset temperature of about 234° C. For example, in some embodiments, AMG176 Form 1 is characterized by DSC as shown in FIG. 3.

[0034] AMG176 Form 1 can be characterized by thermogravimetric analysis (TGA). Thus, AMG176 Form 1 can be characterized by a weight loss in the range of about 5.3% with an onset temperature of about 234° C. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as shown in FIG. 4, where "substantially" means that the reported TGA characteristics may vary by ±5° C.

[0035] AMG176 Form 1 can be characterized by a moisture sorption profile. For example, in some embodiments, AMG176 Form 1 is characterized by a moisture sorption profile as shown in Figure 5, which shows that AMG176 Form 1 is non-hygroscopic up to 95% RH.

[0036] AMG176 Form 1 may be substantially characterized by a single crystal structure as shown in FIG. 6 or as described in the Examples.

[0037] Crystalline Form 2: AMG176 crystalline Form 2 can be characterized by an X-ray powder diffraction pattern obtained as described in the Examples, having peaks at 13.0, 16.9, and 17.3±0.2 degrees 2θ using Cu Kα radiation, and optionally further characterized by additional peaks at 19.1, 21.5, 22.7, 24.3, 27.0, 33.8, 40.0, and 42.8±0.2 degrees 2θ using Cu Kα radiation. In some embodiments, AMG176 Form 2 has an X-ray powder diffraction pattern substantially as shown in FIG. 8, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum are dependent on a number of factors, such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0038] Differential scanning calorimetry (DSC) thermographs were obtained for AMG 176 Form 2 as described in the Examples. The DSC curves show endothermic transitions with onset temperatures at 162°C-169°C, 169°C-173°C, 177°C-179°C, 178°C-182°C, or 232°C-238°C. Thus, in some embodiments, AMG 176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 163°C±3°C, 170°C±3°C, 176°C±3°C, 177°C±3°C, or 238°C±3°C. For example, in some embodiments, AMG 176 Form 2 is characterized by a DSC as shown in FIG. 9, FIG. 10, FIG. 11, FIG. 12, or FIG. 13. In some embodiments, the DSC curve shows an endothermic transition having an onset temperature of 232° C. to 238° C., i.e., AMG 176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 238° C.±3° C. For example, in some embodiments, AMG 176 Form 2 can be characterized by a DSC thermograph as shown in FIG. 9. In some embodiments, the DSC curve shows an endothermic transition having an onset temperature of 177° C. to 179° C., i.e., AMG 176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 176° C.±3° C. For example, in some embodiments, AMG 176 Form 2 can be characterized by a DSC thermograph as shown in FIG. 10. In some embodiments, the DSC curve shows an endothermic transition having an onset temperature of 178° C. to 182° C., i.e., AMG 176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 177° C.±3° C. For example, in some embodiments, AMG176 Form 2 is characterized by DSC as shown in Figure 11. In some embodiments, the DSC curve exhibits an endothermic transition with an onset temperature of 162°C to 169°C, i.e., AMG176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 163°C ± 3°C. For example, in some embodiments, AMG176 Form 2 is characterized by DSC as shown in Figure 12.In some embodiments, the DSC curve exhibits an endothermic transition with an onset temperature of 169° C. to 173° C., i.e., AMG176 Form 2 can be characterized by a DSC thermograph having a transition endotherm with an onset temperature of about 170° C.±3° C. For example, in some embodiments, AMG176 Form 2 is characterized by a DSC as shown in FIG.

[0039] AMG176 Form 2 can be characterized by thermogravimetric analysis (TGA). Thus, AMG176 Form 2 can be characterized by a weight loss in the range of about 12.8% at 140-215°C, about 8.9% at 140-200°C, about 12.7% at 140-190°C, or about 10.1% at 120-180°C. In some embodiments, AMG176 Form 2 can be characterized by a weight loss in the range of about 12.8% at 140-215°C. In some embodiments, AMG176 Form 2 can be characterized by a weight loss in the range of about 8.9% at 140-200°C. In some embodiments, AMG176 Form 2 can be characterized by a weight loss in the range of about 12.7% at 140-190°C. In some embodiments, AMG176 Form 2 can be characterized by a weight loss in the range of about 10.1% at 120-180°C. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as displayed in Figure 10, Figure 11, Figure 12, or Figure 13, where "substantially" means that the reported TGA characteristics may vary by ±5°C. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as displayed in Figure 10. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as displayed in Figure 11. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as displayed in Figure 12. In some embodiments, AMG176 Form 1 has a thermogravimetric analysis substantially as displayed in Figure 13.

[0040] Amorphous AMG176 can be characterized by an X-ray powder diffraction pattern obtained as described in the Examples. In some embodiments, amorphous AMG176 has an X-ray powder diffraction pattern substantially as shown in Figure 14, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum depend on a number of factors, such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0041] Differential scanning calorimetry (DSC) thermographs for amorphous AMG176 were obtained as described in the Examples. The DSC curves show an endothermic transition at 122°C ± 3°C or 163°C ± 3°C. In some embodiments, the DSC curves show an endothermic transition at 122°C ± 3°C. In some embodiments, the DSC curves show an endothermic transition at 163°C ± 3°C. Thus, in some embodiments, amorphous AMG176 can be characterized by a DSC thermograph with a transition endotherm with an onset temperature of 122°C to 130°C or 159°C to 166°C. In some embodiments, amorphous AMG176 can be characterized by a DSC thermograph with a transition endotherm with an onset temperature of 122°C to 130°C. In some embodiments, amorphous AMG176 can be characterized by a DSC thermograph with a transition endotherm with an onset temperature of 159°C to 166°C. For example, in some embodiments, amorphous AMG 176 is characterized by DSC as shown in Figure 15 or Figure 16. In some embodiments, amorphous 176 is characterized by DSC as shown in Figure 15. In some embodiments, amorphous 176 is characterized by DSC as shown in Figure 16.

[0042] Amorphous AMG 176 can be characterized by thermogravimetric analysis (TGA). Thus, amorphous AMG 176 can be characterized by a weight loss of about 3.8% by weight from 28 to 180° C. In some embodiments, anhydrous AMG 176 has a thermogravimetric analysis substantially as displayed in FIG. 17, where "substantially" means that the reported TGA characteristics may vary by ±5° C.

[0043] Amorphous AMG 176 can be characterized by a moisture sorption profile, for example, in some embodiments, amorphous AMG 176 is characterized by a moisture sorption profile as shown in FIG. 18, which shows a weight gain of about 0.95% at 95% RH.

[0044] AMG176 calcium salt hydrate Form 1 may be characterized by an X-ray powder diffraction pattern obtained as described in the Examples, having peaks at 6.8, 7.8, and 15.5±0.2 degrees 2θ using Cu Kα radiation, and optionally further characterized by additional peaks at 11.6, 19.4, and 31.8±0.2 degrees 2θ using Cu Kα radiation. In some embodiments, AMG176 calcium salt hydrate Form 1 has an X-ray powder diffraction pattern substantially as shown in FIG. 20, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum are dependent on a number of factors, such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0045] A differential scanning calorimetry (DSC) thermogram was obtained for AMG176 calcium salt hydrate Form 1 as described in the Examples. The DSC curve shows endothermic transitions at 301° C.±3° C. and 322° C.±3° C. Thus, in some embodiments, AMG176 calcium salt hydrate Form 1 can be characterized by a DSC thermogram having transition endotherms with onset temperatures of 298° C.-304° C. and 319° C.-325° C. For example, in some embodiments, AMG176 calcium salt hydrate Form 1 is characterized by a DSC as shown in FIG. 21.

[0046] AMG176 calcium salt hydrate Form 1 can be characterized by thermogravimetric analysis (TGA). Thus, AMG176 calcium salt hydrate Form 1 can be characterized by a weight loss of about 5.5%, determined to be due to loss of water and ethanol by TGA-IR. In some embodiments, AMG176 calcium salt hydrate Form 1 has a thermogravimetric analysis substantially as displayed in FIG. 21, where "substantially" means that the reported TGA characteristics may vary by ±5° C.

[0047] AMG176 calcium salt hydrate Form 2 can be characterized by an X-ray powder diffraction pattern obtained as described in the Examples having peaks at 6.2, 20.2, and 24.4±0.2 degrees 2θ using Cu Kα radiation, and optionally further characterized by additional peaks at 15.5, 18.2, 19.3, and 21.6±0.2 degrees 2θ using Cu Kα radiation, and / or additional peaks at 10.8, 11.8, 13.9, 16.9, 17.5, 19.6, 22.7, 23.3, 23.5, 25.5, 26.0, and 26.5±0.2 degrees 2θ using Cu Kα radiation. In some embodiments, AMG176 calcium salt hydrate Form 2 has an X-ray powder diffraction pattern substantially as shown in Figure 22, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum depend on a number of factors such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0048] AMG176 calcium salt hydrate Form 2 can be characterized by a moisture sorption profile. For example, in some embodiments, AMG176 calcium salt hydrate Form 2 is characterized by a moisture sorption profile as shown in Figure 23, showing approximately 5.2% weight gain at 70% RH and 23% weight gain at 95% RH.

[0049] AMG176 calcium salt hydrate Form 3 can be characterized by an X-ray powder diffraction pattern obtained as described in the Examples having peaks at 6.2, 6.7, and 8.2±0.2 degrees 2θ using Cu Kα radiation, and optionally further characterized by additional peaks at 17.4, 18.6, and 20.0±0.2 degrees 2θ using Cu Kα radiation, and / or additional peaks at 11.1, 12.0, 13.0, 21.4, 25.4, and 29.4±0.2 degrees 2θ using Cu Kα radiation. In some embodiments, AMG176 calcium salt hydrate Form 3 has an X-ray powder diffraction pattern substantially as shown in Figure 24, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum depend on a number of factors such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0050] A differential scanning calorimetry (DSC) thermogram was obtained for AMG176 calcium salt hydrate Form 3 as described in the Examples. The DSC curve shows an endothermic transition at 314° C.±3° C. Thus, in some embodiments, AMG176 calcium salt hydrate Form 3 can be characterized by a DSC thermogram having a transition endotherm with an onset temperature of 311° C.-317° C. For example, in some embodiments, AMG176 calcium salt hydrate Form 3 is characterized by a DSC as shown in FIG. 25.

[0051] AMG176 calcium salt hydrate Form 3 may be characterized by thermogravimetric analysis (TGA). Thus, AMG176 calcium salt hydrate Form 3 may be characterized by a weight loss of about 3.5%. In some embodiments, AMG176 calcium salt hydrate Form 3 has a thermogravimetric analysis substantially as displayed in FIG. 26, where "substantially" means that the reported TGA characteristics may vary by ±5° C.

[0052] AMG176 calcium salt hydrate Form 3 can be characterized by a moisture sorption profile. For example, in some embodiments, AMG176 calcium salt hydrate Form 3 is characterized by a moisture sorption profile as shown in Figure 27, which shows a weight gain of 7.4% at 95% RH.

[0053] Amorphous AMG176 calcium salt hydrate may be characterized by an X-ray powder diffraction pattern obtained substantially as described in the Examples as shown in Figure 28, where "substantially" means that the reported peaks may vary by ±0.2°. It is well known in the field of XRPD that the relative peak heights in a spectrum depend on a number of factors such as sample preparation and instrument geometry, while the peak positions are relatively insensitive to experimental details.

[0054] For the amorphous AMG176 calcium salt hydrate, a differential scanning calorimetry (DSC) thermogram was obtained as described in the Examples. The DSC curve shows an endothermic transition at 292°C ± 3°C. Thus, in some embodiments, the amorphous AMG176 calcium salt hydrate can be characterized by a DSC thermogram having an endothermic transition with an onset temperature of 289°C to 295°C. For example, in some embodiments, the amorphous AMG176 calcium salt hydrate is characterized by a DSC as shown in Figure 29.

[0055] The amorphous AMG176 calcium salt hydrate can be characterized by thermogravimetric analysis (TGA). Thus, the amorphous AMG176 calcium salt hydrate can be characterized by a weight loss in the range of about 2%. In some embodiments, the amorphous AMG176 calcium salt hydrate has a thermogravimetric analysis substantially as displayed in FIG. 30, where "substantially" means that the reported TGA characteristics may vary by ±5°C.

[0056] The amorphous AMG176 calcium salt hydrate can be characterized by a moisture sorption profile. For example, in some embodiments, the amorphous AMG176 calcium salt hydrate is characterized by a moisture sorption profile as shown in FIG. 31, which shows a weight gain of approximately 9% at 95% RH.

[0057] Pharmaceutical preparations Provided herein are pharmaceutical formulations comprising the crystalline forms disclosed herein and a pharma- ceutically acceptable excipient.

[0058] In some embodiments, the pharmaceutical formulation is in the form of a tablet. In some embodiments, the pharmaceutical formulation is in the form of an immediate release tablet. Solid oral drug compositions (e.g., tablets) or preparations have various release profiles, including immediate release profiles as referred to by FDA guidelines ("Dissolution Testing of Immediate Release Solid Oral Dosage Forms," ​​published August 1997, Section IV-A). In the dissolution testing guidelines for immediate release profiles, a material that dissolves at least 80% in the first 30-60 minutes in solution is considered an immediate release profile. Thus, an immediate release solid dosage form releases most or all of the active ingredient over a short period of time, such as 60 minutes or less, allowing for rapid absorption of the drug. In contrast, sustained release solid oral dosage forms maintain therapeutically effective plasma levels over a similarly long time interval, allowing for the release of the active ingredient over an extended period of time to improve dosing compliance and / or to alter other pharmacokinetic properties of the active ingredient.

[0059] "Pharmaceutically acceptable excipient" refers to a wide range of ingredients that can be combined with the compounds or salts of the present invention to prepare pharmaceutical compositions or formulations. Excipients are additives included in formulations because they either impart or enhance the stability, delivery and manufacturability of the drug product, and are physiologically harmless to the recipient. Regardless of the reason for including an excipient, it is an essential component of the drug product and therefore needs to be safe and well tolerated by the patient. With the teachings and guidance provided herein, one of ordinary skill in the art will easily vary the amount or range of excipients without increasing the viscosity to undesirable levels. Excipients can be selected to achieve the desired bioavailability, the desired stability, resistance to aggregation or degradation or precipitation, protection under freezing, lyophilization or high temperature conditions, or other properties. In general, excipients include, but are not limited to, diluents, colorants, vehicles, anti-adherents, glidants, disintegrants, flavors, coatings, binders, sweeteners, lubricants, adsorbents, preservatives, and the like. Examples of suitable excipients will be well known to those skilled in the art of tablet formulation and can be found, for example, in Handbook of Pharmaceutical Excipients (eds. Rowe, Sheskey & Quinn), 6th edition 2009.

[0060] As used herein, the term "excipient" is intended to refer to, among others, basifying agents, solubilizing agents, glidants, fillers, binders, lubricants, diluents, preservatives, surfactants, dispersing agents, etc. The term also includes substances such as sweetening agents, flavoring agents, coloring agents and preservatives. Such components are generally present in admixture within the tablet.

[0061] Examples of solubilizing agents include, but are not limited to, ionic surfactants (including both ionic and non-ionic surfactants) such as sodium lauryl sulfate, cetyltrimethylammonium bromide, polysorbates (such as polysorbate 20 or 80), poloxamers (such as poloxamer 188 or 207), and macrogols.

[0062] Examples of lubricants, glidants and flow aids include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oils, glyceryl palmitostearate, glyceryl behenate, sodium stearyl fumarate, colloidal silicon dioxide and talc. The amount of lubricant in a tablet can generally be 0.1-5% by weight.

[0063] Examples of disintegrants include, but are not limited to, starch, cellulose, cross-linked PVP, sodium starch glycolate, croscarmellose sodium, and the like.

[0064] Examples of fillers (also known as bulking agents or diluents) include, but are not limited to, starch, maltodextrin, polyols (such as lactose) and cellulose. The tablets provided herein may contain lactose and / or microcrystalline cellulose. Lactose may be used in anhydrous or hydrated form (e.g., monohydrate), and is generally prepared by spray drying, fluidized bed granulation, or roller drying.

[0065] Examples of binders include, but are not limited to, cross-linked PVP, HPMC, microcrystalline cellulose, sucrose, starch, and the like.

[0066] In some embodiments, the pharma- ceutically acceptable excipient may include one or more diluents, binders, or disintegrants. In embodiments, the pharma-ceutically acceptable excipient may include diluents including one or more of microcrystalline cellulose, starch, dicalcium phosphate, lactose, sorbitol, mannitol, sucrose, and methyldextrin, binders including one or more of povidone, hydroxypropylmethylcellulose, hydroxypropylcellulose, and sodium carboxymethylcellulose, and disintegrants including one or more of crospovidone, sodium starch glycolate, and croscarmellose sodium.

[0067] The tablets provided herein may be uncoated or coated (in which case they include a coating). Although uncoated tablets may be used, it is more common to provide coated tablets, in which case a conventional non-enteric coating may be used. Film coatings are known in the art and may be composed of hydrophilic polymeric materials, including but not limited to polysaccharide materials such as hydroxypropylmethylcellulose (HPMC), methylcellulose, hydroxyethylcellulose (HEC), hydroxypropylcellulose (HPC), poly(vinyl alcohol-co-ethylene glycol) and other water-soluble polymers. Although the water-soluble material included in the film coating of the present invention may include a single polymeric material, it may also be formed using a mixture of multiple polymers. The coating may be white or colored, for example gray. Suitable coatings include, but are not limited to, polymeric film coatings (including partially hydrolyzed PVA, titanium dioxide, macrogol 3350, and talc, with optional coloring agents such as iron oxide or indigo carmine or iron oxide yellow or FD&C yellow #6), such as those including polyvinyl alcohol, for example "Opadry® II". The amount of coating is generally 2-4% of the core weight, and in certain specific embodiments 3%. Unless specifically stated otherwise, if the dosage form is coated, it should be understood that references to the weight % of the tablet mean the % of the total tablet, i.e., weight including the coating.

[0068] The pharmaceutical formulations disclosed herein may further comprise a surfactant. As used herein, a surfactant may be cationic, anionic, or nonionic. In some embodiments, the pharmaceutical formulation may comprise a nonionic surfactant. In some embodiments, the surfactant may comprise a polysorbate, a poloxamer, or a combination thereof. In some embodiments, the surfactant may comprise polysorbate 20, polysorbate 60, polysorbate 80, or a combination thereof.

[0069] Method of Treating a Subject Further provided herein is a method of treating a subject suffering from cancer, comprising administering to the subject a therapeutically effective amount of an amorphous or crystalline form disclosed herein, optionally as a pharmaceutical formulation disclosed herein, in some embodiments, the cancer is multiple myeloma, non-Hodgkin's lymphoma, or acute myeloid leukemia.

[0070] Preparation of crystalline forms The crystalline forms disclosed herein can be prepared by various methods known to those skilled in the art. For example, the crystalline forms can be prepared from an amorphous form, a crude form, or another crystalline form of AMG176. In some embodiments, AMG176 is combined with a solvent to form a desired crystalline form, for example as discussed in the examples below. In some embodiments, AMG176 is dissolved in a solvent or combined with a solvent to form a slurry. In some embodiments, AMG176 is combined with a solvent and the solution or slurry so formed is aged to form a crystalline form. In some embodiments, the solution or slurry is heated prior to aging or crystal formation.

[0071] Other embodiments It should be understood that the foregoing description, while being read in conjunction with the detailed description thereof, is intended to be illustrative and not limiting of the scope of the present disclosure, as defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. EXAMPLES

[0072] The following examples are offered for illustrative purposes and are not intended to limit the scope of the invention.

[0073] Materials and Methods Unless otherwise stated, commercially available reagents are used as received without further purification.

[0074] The synthesis of the starting material (AMG176) for the following process is disclosed in U.S. Patent No. 9,562,061. The crystalline forms disclosed herein may be characterized using conventional means, including physical constants and spectral data.

[0075] X-ray powder diffraction: X-ray powder diffraction data were obtained using a PANalytical X-Pert Pro diffractometer. The radiation used was CuKα (1.542 Å) with a voltage and current of 45 kV and 40 mA. Data were collected at ambient temperature from 5.00 to 40.00° 2θ using a step size of 0.0167°. A low background sample holder was used and the stage was rotated with a rotation time of 2.0 seconds. The incident beam path included a 0.02 Radsolar slit, a 15 mm mask, a 4° fixed anti-scatter slit and a programmable divergence slit. The diffracted beam included a 0.02 Radsolar slit, a programmable anti-scatter slit and a 0.02 mm nickel filter.

[0076] Alternatively, XRPD patterns were collected in transmission mode using a PANalytical X'Pert PRO MPD diffractometer with an incident beam of Cu radiation generated using an Optix long microfocus source. An ellipsoidal multilayer mirror was used to focus the Cu Kα X-rays through the sample and onto the detector. Prior to analysis, a silicon standard (NIST SRM 640e) was analyzed to confirm that the observed position of the Si111 peak matched the NIST certified position. Samples were sandwiched between 3 μm thick films and analyzed in transmission geometry. A beam stop, short antiscatter extension, and knife edge were used to minimize background caused by the atmosphere. Soller slits of the incident and diffracted beams were used to minimize spread from axial divergence. Diffraction patterns were collected using a scanning position sensitive detector (X'Celerator) located 240 mm away from the sample, and data collector software v.2.2b. The data collection parameters for each pattern are shown above the images in the data section of this report, and include a divergence slit (DS) in front of the mirror.

[0077] XRPD parameters were collected in reflection mode on a PANalytical X'Pert PRO MPD diffractometer using an incident beam of Cu Kα radiation generated using a long micropoint source and a nickel filter. The diffractometer was configured to use a symmetric Bragg-Brentano geometry. Prior to analysis, a silicon standard (NIST SRM 640e) was analyzed to confirm that the observed position of the Si111 peak matched the NIST certified position. Sample specimens were packed into wells. Anti-scatter slits (SS) were used to minimize background caused by the atmosphere. Soller slits of the incident and diffracted beams were used to minimize spread from axial divergence. Diffraction patterns were collected using a scanning position sensitive detector (X'Celerator) positioned 240 mm from the sample and data collector software v.2.2b. Data acquisition parameters for each pattern are displayed in the images above in the data section of this report, including the divergence slit (DS) and incident beam SS.

[0078] In the figures referencing a given indexed XRPD pattern, agreement between the accepted peak positions marked with red bars and the observed peak positions indicates a consistent unit cell determination. Successful indexing of a pattern indicates that the sample is composed primarily of a single crystalline phase unless otherwise noted. The space group consistent with the assigned annihilation symbols, unit cell parameters, and derived quantities is labeled in the figure below. To confirm the tentative indexing solution, the molecular packing motifs within the crystalline unit cell must be determined. No attempt at molecular packing was made.

[0079] Single crystal structure: A colorless needle-like crystal (monohydrate) with dimensions of 0.18 x 0.11 x 0.05 mm was mounted on a nylon loop using a very small amount of paratone oil. The crystal was mounted and the lattice was determined to be monohydrate, then the temperature was raised to 400°K to dehydrate. The crystal was left at 400°K for 4 hours, then the temperature was lowered to 173°K to collect the data. The data was collected using a Bruker CCD (charge-coupled device)-based diffractometer equipped with an Oxford Cryostream cryostat operating at 173°K. The data was measured for 30 seconds using omega and phi scans of 0.5° per frame. The total number of images was based on the results from the program COSMO, with a redundancy of 4.0 and a completeness of 0.83 Å, expected to be 100%. Lattice parameters were obtained using APEX II software and refined using SAINT for all observed reflections. Data reduction was performed using SAINT software. Scaling and absorption corrections were applied using the SADABS multiscan technique. The structure was solved by direct methods using the SHELXS-97 program and refined by least-squares fitting to F2, SHELXL-97, implemented in SHELXTL-PC V6.10.

[0080] Differential Scanning Calorimetry: Differential scanning calorimetry (DSC) analysis was performed on a TA Instruments Q100 instrument. Approximately 1 mg sample size was weighed into a standard aluminum DSC pan, and the pan was not crimped. The sample was heated from ambient temperature to 300° C. at 10° C. / min under 50 mL / min dry nitrogen. Temperature modulated DSC analysis was performed using a TA Instruments Q100 instrument. Approximately 1 mg sample size was used in an aluminum uncrimped pan. The sample was equilibrated at 20° C., held for 5 minutes, and then heated to 300° C. at a heating rate of 3° C. / min under 50 mL / min dry nitrogen. The modulated temperature was ±0.75° C. every 45 seconds.

[0081] Alternatively, DSC was performed using a TA Instruments Q2000 Differential Scanning Calorimeter. Temperature calibration was performed using NIST tracked indium metal. Samples were placed in aluminum Tzero pans, covered with lids, and weights were accurately recorded. A weighing aluminum pan configured as the sample pan was placed on the reference side of the cell. Data collection parameters and pan configuration for each thermogram are displayed in images in the data section of this report. Thermogram method codes are abbreviations for the start and end temperatures and heating rate, e.g., -30-250-10 means "-30°C to 250°C at 10°C / min."

[0082] Alternatively, MDSC data were obtained on a TA Instruments Q2000 Differential Scanning Calorimeter equipped with a refrigerated cooling system (RCS). Temperature calibration was performed using NIST tracked indium metal. Samples were placed in aluminum DSC pans and the weights were accurately recorded. The pans were covered with lids and the lids were crimped. The weighed and crimped aluminum pans were placed in the reference side of the cell. Data were obtained using a modulated temperature amplitude of ±0.8°C and a duration of 60 seconds with an elementary heating rate of 2°C / min from -30°C to 250°C. The reported glass transition temperatures are obtained from the inflection point of the step change in the reversing heat flow versus temperature curve.

[0083] Alternatively, DSC was performed using a Mettler-Toledo DSC3+ differential scanning calorimeter. Temperature calibration was performed using adamantane, phenyl salicylate, indium, tin, and zinc. Samples were placed in hermetically sealed or open aluminum DSC pans and the weights were accurately recorded. A weighed aluminum pan configured as the sample pan was placed on the reference side of the cell. Samples were analyzed from -30 to 250°C at a ramp rate of 10°C / min. Thermograms are plotted by reference temperature (x-axis), but results are reported according to sample temperature.

[0084] Thermal Analysis: Thermogravimetric analysis was performed on a TA Instruments Q500 instrument. Sample sizes of approximately 1-5 mg were used in aluminum pans. Samples were heated at 10°C / min from ambient temperature to 400°C under 25 mL / min dry nitrogen.

[0085] Alternatively, TG analysis was performed using a TA Instruments Q5000 IR Thermogravimetric Analyzer. Temperature calibration was performed using nickel and Alumel™. Each sample was placed in an aluminum pan. The samples were hermetically sealed, the lid was drilled, and then inserted into the TG furnace. The furnace was heated under nitrogen. Data acquisition parameters for each thermogram are displayed in the images in the data section of this report. Thermogram method codes are abbreviations for the start and end temperatures and heating rate, e.g., 25-350-10 means "25°C to 350°C at 10°C / min."

[0086] Alternatively, thermogravimetric analysis was performed using a Mettler Toledo TGA / DSC3+ analyzer. Temperature calibration was performed using phenyl salicylate, indium, tin, and zinc. Samples were placed on aluminum plates. Open pans were inserted into the TG furnace. The furnace was heated under nitrogen. Each sample was heated from ambient to 350° C. at a ramp rate of 2, 5, or 10° C. / min. Thermograms are plotted by reference temperature (x-axis), but results are reported according to sample temperature.

[0087] Hygroscopicity: Hygroscopicity data was collected at 25°C using a VTI vapor sorption analyzer. Sample sizes of approximately 4-10 mg were used on standard platinum pans. Hygroscopicity was evaluated from 5 to 95% RH in 5% RH increments. Data was collected for adsorption and desorption cycles. Equilibrium criteria were set at 0.001% weight change in 10 minutes with a maximum equilibration time of 180 minutes.

[0088] NMR: Proton solution NMR spectra were obtained on a Varian UNITYINOVA-400 spectrophotometer at 25° C. by Spectral Data Services of Champaign, Ill. Samples were dissolved in DMSO-d6. In some cases, solution NMR spectra were obtained on an SSCI using an Agilent DD2-400 spectrophotometer using deuterated DMSO or methanol.

[0089] 13 C SSNMR data was obtained from 600MHz ( 1 H. All experiments were performed using a Bruker DSX spectrophotometer operating at 100 Hz. A 4 mm H / F / X spinning probe operating at a spinning frequency of 14 kHz was used. A CPMAS with the TOSS program was used with a recycle delay of 10 seconds. A 2.5 μs 1 H 90° pulse and 8μs 13 A C 180° pulse was used. Decoupling was performed using a spinal64 sequence. 4096 transients were acquired for signal averaging. Data were processed with Topspin 3.0 software.

[0090] Example 1: AMG176 Form 1 AMG176 Form 1 has acceptable pharmaceutical and physical properties, is scalable on a kilogram scale, and can be compounded during pharmaceutical formulation as the sodium salt in situ to achieve the desired drug concentration to enable IV delivery. AMG176 can also be converted to amorphous material to aid oral administration, if desired, depending on the dosage range. Dog PK studies suggested that absorption was improved by only 10-fold when AMG176 was administered as an amorphous suspension, compared to a crystalline suspension. Although improved exposure was observed by administering the amorphous material, this exposure would likely plateau if gram-level doses were required for oral delivery.

[0091] AMG176 (121 g) was dissolved in ethyl acetate. Ethanol (800 mL) was added and the combination was mixed with heating for 20 minutes. Water (250 mL) was added dropwise over 30 minutes. The slurry was cooled to room temperature for 2 hours and then further cooled in an ice bath for 4 hours before filtering. The wet cake was washed with cold 30% ethanol in water (300 mL). The cake was air dried for 2 days and further dried under vacuum at 40° C. for 4 days. The dried solid was identified as crystalline AMG176 Form 1.

[0092] AMG176 Form 1 consists of anhydrous / unsolvated AMG176 and is likely thermodynamically stable between 2 and 79 °C. The XRPD pattern for Form 1 was successfully indexed, indicating that the material consists primarily or exclusively of a single crystalline phase. The unit cell volume obtained from the indexing solution contains the minimum free volume consistent with anhydrous / unsolvated AMG176. The proton NMR spectrum for Form 1 was measured in deuterated DMSO. The spectrum was consistent with the chemical structure of AMG176.

[0093] AMG176 Form 1 was analyzed by DSC and TGA. Negligible weight loss was observed by TGA up to 220° C., consistent with anhydrous / nonsolvated material (FIG. 4). The DSC thermogram was unremarkable until a sharp endotherm occurred at 232° C., likely corresponding to simultaneous melting and decomposition (FIG. 3).

[0094] AMG176 Form 1 was also analyzed by DVS (Figure 5). The material exhibited low hygroscopicity, absorbing only 0.3% water vapor from 5% to 95% RH. All of this weight was lost upon desorption, and virtually no hysteresis was observed. XRPD of the material after DVS showed no morphological changes (Figure 7).

[0095] [Table 1]

[0096] Single crystal data: The crystal structure of AMG176 form 1 has been determined. The crystal parameters are summarized in Table 2 and the unit cell is displayed in Figure 6.

[0097] [Table 2]

[0098] Solid state stability evaluation of Form 1 of AMG 176 was performed at accelerated conditions of 25° C. / 60% RH (open), 40° C. (closed), 60° C. (closed), and 40° C. / 75% RH (open) to determine the chemical and physical stability of AMG 176. After 8 weeks, Form 1 was physically and chemically stable under all stress conditions, as displayed in FIG.

[0099] Example 2: AMG176 Form 2 AMG176 Form 2 consists of an isostructural solvate containing one mole of 2-MeTHF, THF, MTBE, or 1,4-dioxane per mole of AMG176. This form was produced from multiple slurry or steam stress experiments in relevant solvent systems. Representative samples from each solvent were characterized for comparison by XRPD indexing, proton NMR, DSC, and TGA. The solvent systems in which AMG176 Form 2 was produced were IPA / heptane, 2-MeTHF, THF / water, MTBE, and 1,4-dioxane / water.

[0100] Peak shifts consistent with isostructural solvates were observed in the XRPD patterns. Unit cell volumes from the XRPD indexing solutions and proton NMR spectra were all consistent with approximately 1 mole of each solvent in the crystal lattice. Additionally, the amount of weight loss in each of the TGA thermograms is consistent with a loss of 0.8 to 1 mole of each solvent. The desolvation events occurred at slightly different onset temperatures based on the endotherms observed by DSC, and overlays of DSC and TGA thermograms for each sample are presented in Figures 9-13 to illustrate the differences in weight loss upon heating. The DSC thermograms for the samples from 2-MeTHF and 1,4-dioxane exhibit an endothermic event concomitant with the desolvation endotherm, likely indicative of form conversion.

[0101] Based on the DSC and TGA data, experiments were designed to investigate the desolvation of various samples of AMG176 Form 2 by heating, vacuum, and VT-XRPD. Attempts to desolvate samples of Form 2 under vacuum at about 70° C. and heating at about 164-165° C. for 10 minutes were insufficient to remove the solvent.

[0102] Attempts to reproduce form 2 by slurrying in 2-MeTHF on a 600 mg scale were successful. This material was utilized for form screening experiments.

[0103] For example, AMG176 Form 2 was formed by slurrying AMG176 in IPA / heptane (1:1, 8.4 mg / mL) for 8 hours at 25° C. The solvent was evaporated. The remaining solid was identified as crystalline Form 2 by XRPD.

[0104] [Table 3]

[0105] A summary of the XRPD peak distinctions between AMG Form 1 and Form 2 is provided in Table 4 below.

[0106] [Table 4]

[0107] Example 3: Amorphous AMG176 Numerous evaporation and vapor diffusion experiments during morphology screening yielded amorphous material. Noting the ease of preparation through fast evaporation, material was prepared on a ∼500 mg scale from DCM for use in additional morphology screening experiments and characterization.

[0108] The amorphous material was characterized by proton NMR, DVS, temperature modulated DSC, and TGA. The proton NMR spectrum was consistent with the chemical structure of AMG176.

[0109] The DVS isotherm is shown in Figure 18. The amorphous material exhibited limited hygroscopicity, absorbing 0.95 wt% water vapor from 5% to 95% RH. More weight was lost (1.59%) than gained upon adsorption, likely due to loss of residual DCM and / or water from the starting material. The XRPD of the material after DVS was consistent with an amorphous material.

[0110] The observation of a glass transition (Tg) can be characteristic of the amorphous nature of a material. Temperature modulated DSC (mDSC) was performed to determine the Tg of a material and is shown in FIG. 15. mDSC allows the total heat flow signal to be separated into thermodynamic (heat capacity) and kinetic components. Thus, the Tg can typically be seen as a step change in the inversion signal. Amorphous AMG176 exhibits a Tg at approximately 163° C. (ΔCP: 0.3 J / (g° C.)). It should be noted that differences in solvent and / or water content in the sample can shift the temperature at which the glass transition occurs.

[0111] The TGA thermogram is shown in Figure 17. A gradual weight loss of 3.8 wt % was observed from 28 to 180°C, consistent with loss solvent (presumably DCM and water).

[0112] Attempts to crystallize the amorphous material were set up by steam stressing and slurrying under various conditions. The majority of the experiments led to crystallization to Form 1, except for those from related solvent systems (1,4-dioxane / water, 2-MeTHF, and MTBE) that led to Form 2. Only one experiment, steam stress with water at room temperature, did not lead to crystallization, likely due to lack of sufficient solubility.

[0113] For example, amorphous AMG176 was formed by dissolving AMG176 (3.9 g) in a minimum amount of EtOAc and then rapidly precipitating with heptane. The suspension was filtered. The remaining solid was identified as amorphous by XRPD.

[0114] Solid state stability evaluation of AMG 176 amorphous material and Form 1 was performed at 25° C. / 60% RH (open), 40° C. (closed), 60° C. (closed), and 40° C. / 75% RH (open) accelerated conditions to determine the chemical and physical stability of AMG 176. After 8 weeks, the amorphous material and Form 1 were physically and chemically stable under all stress conditions, as displayed in FIG.

[0115] Example 4: AMG176 Calcium Hydrate Form 1 AMG176 sodium salt was dissolved in ethanol at 140 mg / mL to which 0.55 equivalents of CaCl2.2H2O was added as a solution in ethanol resulting in a precipitate. The suspension was filtered and then a 0.5 mL aliquot of the filtrate was layered with heptane and then allowed to stand at room temperature to crystallize. The solid was identified as crystalline by XRPD (Figure 20).

[0116] TGA analysis showed a weight loss of approximately 5.5%, which was confirmed by TGA-IR to be due to loss of water and ethanol. Approximately 0.5 molar equivalents of Ca were detected. The Ca salt appeared to melt at approximately 301° C., recrystallize, and undergo further melting at 322° C. before decomposing. The TGA and DSC data are presented in FIG. 21.

[0117] [Table 5]

[0118] AMG176 Calcium Hydrate Form 1- 1 H NMR confirmed salt formation with residual ethanol. TGA analysis showed approximately 5.5% weight loss from 25 to 225 °C, which was confirmed by TGA-IR to be due to loss of water and ethanol. Approximately 0.5 molar equivalents of Ca were detected by CE. 1 H NMR(400MHz,DMSO-d6)δ ppm 0.78-1.04(m,4H)1.04-1.18(m,4H)1.18-1.29(m,1H)1.29-1.53(m,2H)1.54-1.79(m,4H)1.84(br d,J=7.67Hz,3H)1.91-2.11(m,4H)2.22-2.49(m,3H)2.65-2.83(m,2H)2.89-3.07(m,1H)3.07-3.12(m,3H)3.17(br d,J=14.10Hz,1H)3.34-3.60(m,6H)3.64-3.82(m,2H)3.82-4.06(m,5H)4.35(t,J=5.08Hz,1H)5.36(br dd,J=15.45,9.23Hz,2H)6.00(br s,2H)6.73(d,J=8.09Hz,2H)6.88-6.96(m,2H)7.01(dd,J=7.98,1.76Hz,2H) 7.16(d,J=2.28Hz,2H)7.27(dd,J=8.50,2.28Hz,2H)7.69(d,J=8.71Hz,1H).

[0119] Example 5: AMG176 Calcium Hydrate Form 2 AMG176 sodium was dissolved in EtOAc at 43.6 mg / mL, to which was added 0.55 equivalents of CaCl2.2H2O as a solution in ethanol. The filtrate was seeded with AMG176 calcium Form 1 and then stirred at room temperature. The solid was identified as crystalline by XRPD (Figure 22).

[0120] AMG176Ca salt Form 2 had a weight loss of approximately 3.6% by TGA (Figure 30), consistent with the presence of 3.2% water. Moisture sorption analysis showed a weight gain of approximately 5.2% at 70% RH and 23% at 95% RH, likely due to the presence of residual NaCl (Figure 23). The Ca content was determined to be 3.17%, which corresponds to approximately 0.5 moles of Ca. The water solubility of the crystalline salt was determined to be approximately 24 μg / mL.

[0121] [Table 6]

[0122] AMG176 Calcium Hydrate Form 2- 1 H NMR confirmed salt formation with residual ethanol. TGA from 25 to 225 °C showed a weight loss of 3.6%, which was confirmed to be water (3.2%) by KF. The Ca content was determined by capillary electrophoresis to be 3.17%, which corresponds to 0.5 moles of calcium. 1 H NMR(400MHz,DMSO-d6)δ ppm 0.85(d,J=6.01Hz,3H)0.99-1.21(m,5H)1.24-1.44(m,1H)1.55-1.79(m,3H)1.84(br d,J=7.46Hz,3H)1.91-2.09(m,4H)2.22-2.48(m,2H)2.65-2.83(m,2H)2 .89-3.07(m,1H)3.09(s,2H)3.13-3.31(m,1H)3.34-3.60(m,4H)3.72(br d,J=14.93Hz,1H)3.85-4.03(m,3H)4.35(t,J=5.08Hz,1H)5.36(dd,J=15.34,9.12Hz,1H)6.01(br dd,J=15.65,8.40Hz,1H)6.72(d,J=8.09Hz,1H)6.91-6.97(m,1H)7.01(dd,J=7.98,1. 76Hz,2H)7.16(d,J=2.28Hz,2H)7.27(dd,J=8.50,2.28Hz,2H)7.70(d,J=8.71Hz,1H).

[0123] Example 6: AMG176 Calcium Hydrate Form 3 AMG176 calcium salt Form 2 was stirred in water at 17.5 mg / mL for 24 hours at 25° C. The solid was characterized as crystalline by XRPD.

[0124] Upon TGA analysis, there was a weight loss of approximately 3.5% (Figure 26), likely the result of water loss based on a KF of 4.3% water. A melting point of approximately 314°C was observed by DSC analysis (Figure 25). The material was determined to be hygroscopic, absorbing approximately 7.4% water at 95% RH (Figure 27). The amount of Ca present in the sample was approximately 1.8%, which was lower than expected.

[0125] [Table 7]

[0126] AMG176 Calcium Hydrate Form 3- 1 H NMR spectrum confirmed no salt formation and no residual organic solvents. The water content was 4.3% by KF. 1H NMR(400MHz,DMSO-d6)δ ppm 0.85(br d,J=6.01Hz,3H)1.13(d,J=7.05Hz,3H)1.36(br t,J=9.85Hz,1H)1.55-1.79(m,3H)1.84(br d,J=7.46Hz,2H)1.92-2.03(m,3H)2.21-2.48(m,2H)2.65-2.83(m,2H)2 .89-3.07(m,1H)3.09(s,2H)3.13-3.31(m,1H)3.44-3.62(m,2H)3.72(br d,J=14.72Hz,2H)3.86-4.06(m,3H)5.36(br dd,J=15.24,9.02Hz,2H)6.01(br dd,J=15.13,8.09Hz,1H)6.72(d,J=8.09Hz,1H)6.92-6.97(m,1H)7.01(dd,J=8.09,1. 66Hz,1H)7.16(d,J=2.49Hz,1H)7.27(dd,J=8.50,2.49Hz,1H)7.70(d,J=8.50Hz,1H).

[0127] Example 7: Amorphous AMG176 calcium hydrate Amorphous AMG 176 calcium hydrate appeared by microscopy as irregularly shaped particles with some minor birefringence present. When analyzed by XRPD, the amorphous material did not exhibit any characteristic diffraction peaks (Figure 28). Approximately 2% weight loss was observed by TGA (Figure 30), likely due to residual solvent. DSC analysis showed a melting endotherm at approximately 292°C (Figure 29). The amorphous material likely recrystallized during the analysis even though no distinct recrystallization endotherm was observed. During moisture sorption analysis, the amorphous material was determined to be hygroscopic with approximately 9% weight gain at 95% RH (Figure 31).

[0128] The amorphous material exhibited limited hygroscopicity, absorbing 0.95 wt.% water vapor from 5% to 95% RH. More weight was lost (1.59%) than gained upon adsorption, likely due to loss of residual DCM and / or water from the starting material. XRPD of the material after DVS was consistent with amorphous material.

[0129] The foregoing descriptions are set forth merely for clarity of understanding, and no unnecessary limitations should be understood therefrom, since modifications within the scope of the invention may be apparent to those skilled in the art.

[0130] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integers or steps or group of integers or steps.

[0131] Throughout this specification, when a composition is described as comprising ingredients or raw materials, it is understood that the composition can also consist essentially of, or consist of, any combination of the listed ingredients or raw materials, unless otherwise stated. Similarly, when a method is described as comprising particular steps, it is understood that the method can also consist essentially of, or consist of, any combination of the listed steps, unless otherwise stated. The invention illustratively disclosed herein may suitably be practiced in the absence of any element or step not specifically disclosed herein.

[0132] As will be apparent to one of ordinary skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope and spirit of the present disclosure. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0133] The implementation of the methods disclosed herein, and their individual steps, can be performed manually and / or with the aid of automation provided by electronic equipment. Although the methods have been described with respect to specific embodiments, one skilled in the art will readily appreciate that other ways of performing the acts associated with the methods can be used. For example, the order of various steps can be altered without departing from the scope or spirit of the methods, unless otherwise noted. In addition, some of the individual steps can be combined, omitted, or further subdivided into additional steps.

[0134] In the context of this disclosure (especially in the context of the claims), the use of the terms "a," "an," "the," and similar referents should be construed to include both the singular and the plural, unless otherwise specified. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise specified herein, and each separate value is incorporated herein as if it were individually recited herein. The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended to better illustrate the disclosure herein and is not a limitation on the scope of the disclosure herein, unless otherwise specified. No language herein should be construed as indicating any non-claimed element as essential to the practice of the disclosure herein.

[0135] All patents, publications, and references cited herein are hereby incorporated by reference in their entirety. In the event of a conflict between the present disclosure and the incorporated patents, publications, and references, the present disclosure shall control.

Claims

1. The crystalline form of AMG176 is solid at concentrations of 12.39, 19.40, 20.46, 27.24, 28.07, 30.54, 33.09, 33.80, 37.18, 41.80, 42.98, 54.86, 58.69, 60.01, 63.11, 80.06, 85.77, 117.20, 119.77, 120.86, 127.04, 129.01, 129.71, 131.30, 132.24, 133.58, 139.16, 140.11, 140.69, 152.08, and 169.82 ± 0.5 ppm. １３ Crystal morphology characterized by 13C NMR peaks ("AMG176 morphology 1").

2. The crystalline morphology according to claim 1, further characterized by XRPD pattern peaks at 14.2, 18.0, and 18.6 ± 0.2° 2θ using Cu Kα radiation.

3. The crystalline morphology according to claim 2, further characterized by XRPD pattern peaks at 12.6, 17.0, 22.7, and 26.6 ± 0.2° 2θ using Cu Kα radiation.

4. The crystalline morphology according to claim 3, further characterized by XRPD pattern peaks at 5.6, 12.3, 12.8, 14.5, 16.7, 24.4, 25.2, 27.1, 28.3, and 28.9 ± 0.2° 2θ using Cu Kα radiation.

5. The crystalline morphology according to claim 1, having an endothermic transition between 233°C and 238°C, as measured by differential scanning calorimetry.

6. The crystalline morphology of AMG176 ("AMG176 morphology 2") is characterized by XRPD pattern peaks at 13.0, 16.9, and 17.3 ± 0.2° 2θ using Cu Kα radiation.

7. The crystalline morphology according to claim 6, further characterized by XRPD pattern peaks at 19.1, 21.5, 22.7, 24.3, 27.0, 33.8, 40.0, and 42.8 ± 0.2° 2θ using Cu Kα radiation.

8. The crystalline morphology according to claim 6, having an endothermic transition between 232°C and 238°C, as measured by differential scanning calorimetry.

9. The crystalline form according to claim 6, having an endothermic transition at 162°C to 169°C, 169°C to 173°C, 177°C to 179°C, or 178°C to 182°C, as measured by differential scanning calorimetry.

10. The crystalline form according to claim 6, wherein the endothermic transition occurs at 163°C ± 3°C, 170°C ± 3°C, 176°C ± 3°C, or 177°C ± 3°C.

11. The amorphous form of AMG176 ("AMG176 amorphous") substantially has an XRPD pattern as shown in Figure 14.

12. The amorphous form according to claim 11, having an endothermic transition between 122°C and 130°C or 159°C and 166°C, as measured by differential scanning calorimetry.

13. The amorphous form according to claim 12, wherein the endothermic transition occurs at 122°C or 163°C ± 3°C.

14. A crystalline form of AMG176 as a calcium salt hydrate, characterized by XRPD pattern peaks at 6.8, 7.8, and 15.5 ± 0.2° 2θ using Cu Kα radiation ("AMG176 calcium hydrate form 1").

15. The crystalline morphology according to claim 14, further characterized by XRPD pattern peaks at 11.6, 19.4, and 31.8 ± 0.2° 2θ using Cu Kα radiation.

16. The crystalline morphology according to claim 14, having endothermic transitions at 301°C ± 3°C and 322°C ± 3°C, as measured by differential scanning calorimetry.

17. A crystalline form of AMG176 as a calcium salt hydrate, characterized by XRPD pattern peaks at 6.2, 20.2, and 24.4 ± 0.2° 2θ using Cu Kα radiation ("AMG176 calcium hydrate form 2").

18. The crystalline morphology according to claim 17, further characterized by XRPD pattern peaks at 15.5, 18.2, 19.3, and 21.6 ± 0.2° 2θ using Cu Kα radiation.

19. The crystalline morphology according to claim 17, further characterized by XRPD pattern peaks at 10.8, 11.8, 13.9, 16.9, 17.5, 19.6, 22.7, 23.3, 23.5, 25.5, 26.0, and 26.5 ± 0.2° 2θ using Cu Kα radiation.

20. A crystalline form of AMG176 as a calcium salt hydrate, characterized by XRPD pattern peaks at 6.2, 6.7, and 8.2 ± 0.2° 2θ using Cu Kα radiation ("AMG176 calcium hydrate form 3").

21. The crystalline morphology according to claim 20, further characterized by XRPD pattern peaks at 17.4, 18.6, and 20.0 ± 0.2° 2θ using Cu Kα radiation.

22. The crystalline morphology according to claim 20, further characterized by XRPD pattern peaks at 11.1, 12.0, 13.0, 21.4, 25.4, and 29.4 ± 0.2° 2θ using Cu Kα radiation.

23. The crystalline morphology according to claim 20, having an endothermic transition at 314°C ± 3°C, as measured by differential scanning calorimetry.

24. An amorphous form of AMG176 as a calcium salt hydrate, substantially possessing an XRPD pattern as shown in Figure 28 ("AMG176 amorphous calcium hydrate").

25. The crystalline morphology according to claim 24, having an endothermic transition at 292°C ± 3°C as measured by differential scanning calorimetry.

26. A pharmaceutical preparation comprising a crystalline or amorphous form according to any one of claims 1 to 25 and a pharmaceutically acceptable excipient.

27. A method for treating a subject suffering from cancer, comprising administering to the subject a therapeutically effective amount of a crystalline or amorphous form described in any one of claims 1 to 25.

28. The method according to claim 27, wherein the cancer is multiple myeloma, non-Hodgkin lymphoma, or acute myeloid leukemia.

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