WO2022036269A1

WO2022036269A1 - CRYSTALLINE FORMS OF AN HIF-1α PROLYL HYDROXYLASE INHIBITOR

Info

Publication number: WO2022036269A1
Application number: PCT/US2021/046021
Authority: WO
Inventors: Maurice MARSINI; Nicole White; Richard WINNIKE
Original assignee: Gb004, Inc.
Priority date: 2020-08-13
Filing date: 2021-08-13
Publication date: 2022-02-17

Abstract

Crystalline free base and salt forms of a HIF-1α prolyl hydroxylase inhibitor are provided, as well as pharmaceutical compositions comprising the same. Also provided are methods for their production and use in the context of treating conditions for which inhibition of HIF-1α prolyl hydroxylase is desired.

Description

CRYSTALLINE FORMS OF AN HIF-1α PROLYL HYDROXYLASE INHIBITOR

BACKGROUND

Technical Field

The invention relates to crystalline free base and salt forms of a HIF-1α prolyl hydroxylase, as well as to pharmaceutical compositions comprising the same, and to methods for their production and use in the context of treating conditions for which inhibition of the HIF-1α prolyl hydroxylase is desired.

Description of the Related Art

Inflammatory bowel disease (IBD) is characterized by repeated wounding and inflammation of the intestinal mucosa and typically involves a breach in intestinal barrier integrity, which allows the influx of luminal antigens and sets up a vicious cycle of inflammation and epithelial injury. During mucosal inflammation, changes in metabolic activity and vascular tissue damage lead to a reduction in tissue-oxygen tension (hypoxia), in which healing processes such as angiogenesis, cell migration, and re-epithelialization occur. Typically it is the epithelial cells on the surface of mucosal tissue that enter an induced state of hypoxia due to the presence of inflammation and the body's response to this hypoxic condition is to increase the presence of hypoxia inducible factor-1 alpha (HIF-1α), which subsequently drives the expression of downstream HIF-1 target genes, inter alia, erythropoietin. As such, HIF-1α is an important mediator in the body's response to inflammation.

The cellular concentration of HIF-1α is regulated by prolyl hydroxylase (PHD) enzymes that serve to destabilize HIF-1α during periods of normoxia resulting in the destruction of this protein. Inhibition of HIF-1α prolyl hydroxylase thus leads to increased stabilization of HIF-1α resulting in a up regulation of HIF-1α which in turn leads to a corresponding increased response to inflammation. In subjects suffering from one or more inflammatory epithelial diseases, treatment with effective HIF-1α prolyl hydroxylase inhibitors can increase the level of the body's cellular inflammatory response. In addition, during periods of low level inflammation in the case of chronic diseases, HIF-1α prolyl hydroxylase inhibitors can increase the amount of epithelial cell healing over that which the body would normally provide. While advances have been made with regard to inhibition of HIF-1α prolyl hydroxylases, there remains a need in the art for inhibition of HIF-1α prolyl hydroxylases, as well as the need to treat various conditions and/or disorders that would benefit from the same. The present invention fulfills that need and provides further related advantages as evident in the following disclosure.

BRIEF SUMMARY

In brief, crystalline free base and salt forms of a HIF-1α prolyl hydroxylase, as well as to pharmaceutical compositions comprising the same, and to methods for their production and use in the context of treating conditions for which inhibition of the HIF-1α prolyl hydroxylase is desired, are provided.

Accordingly, in some embodiments, the HC1 salt of tert-butyl 4-[[l-[(4- chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate is provided in crystalline form.

In some embodiments, the free base of tert-butyl 4-[[l-[(4-chlorophenyl)methyl]- 3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate is provided in crystalline form.

In some embodiments, the free base of l-(4-chlorobenzyl)-3-hydroxy-4- (piperazin-l-ylmethyl)pyridin-2(lH)-one is provided in crystalline form.

In some embodiments, the napsylate salt of tert-butyl 4-[[l-[(4- chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate is provided in crystalline form.

In some embodiments, the xinafoate salt of tert-butyl 4-[[l-[(4- chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate is provided in crystalline form.

In some embodiments, a pharmaceutical composition is provided comprising a crystalline form of tert-butyl 4-[[l-[(4-chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate in salt or free base form and a pharmaceutically acceptable carrier.

In some embodiments, a method for inhibiting HIF-1α prolyl hydroxylase activity is provided comprising contacting the HIF-1α prolyl hydroxylase with an effective amount of a crystalline form of tert-butyl 4-[[l-[(4-chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate in salt or free base form. In some embodiments, a method for treating a disease or condition for which HIF- lα prolyl hydroxylase inhibition is beneficial is provided, comprising administering to a subject in need thereof an effective amount of a crystalline form of tert-butyl 4-[[l-[(4- chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate in salt or free base form.

In some embodiments, a method for treating an inflammatory disease is provided, comprising administering to a subject in need thereof effective amount of a crystalline form of tert-butyl 4-[[l-[(4-chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1- carboxylate in salt or free base form.

BRIEF DESCRIPTION THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

Figure 1 shows the relationship between various crystal forms of Compound (1) Free Base and HC1 salt.

Figure 2 shows an XRPD 29 diffractogram of Compound (1) Free Base.

Figure 3A shows an XRPD spectrum of the free base Form of Compound (1) and Figure 3B shows the XRPD spectrum of Form E of the HC1 salt of Compound (1).

Figure 4 shows an ¹H NMR Spectrum of Compound (1) Free Base (500.12 MHz, d6-DMSO).

Figure 5 shows a DSC thermogram of Compound (1) Free Base showing the melting transition.

Figure 6 shows a DVS isotherm plot of Compound (1) Free Base.

Figure 7 shows an FT-IR spectrum of Compound (1) Free Base.

Figure 8 shows a view of an asymmetric unit with one molecule of Compound (1) Free Base crystal structure, with the structural formula below (non-H atoms labeled).

Figure 9A shows a simulated XRPD 29 diffractogram of Compound (1) Free Base at 199 K and Figure 9B shows an experimental XRPD 29 diffractogram of Compound (1) Free Base at 298 K.

Figure 19 shows an XRPD 29 diffractogram of Compound (2). Figure 11 shows a TG/DTA thermogram of Compound (2).

Figure 12 shows an XRPD 2θ diffractogram of Compound (1) HC1 salt Form A.

Figure 13 shows a TG/DTA thermogram of Compound (1) HC1 salt Form A.

Figure 14A shows a DSC thermogram of the first heating cycle from 20 °C- 150 °C of Compound (1) HC1 salt Form A. Figure 14B shows a DSC thermogram of the first cooling cycle from 150 °C-20 °C of Compound (1) HC1 salt Form A. Figure 14C shows a second heating cycle from 20 °C-300 °C of Compound (1) HC1 salt Form A.

Figure 15A shows a¹H NMR spectrum (500.12 MHz, DMSO-d6) of Compound (1) HC1 salt Form A. Figure 15B shows a heteronuclear single quantum correlation (HSQC) of Compound (1) HC1 salt Form A.

Figure 16 shows a GVS isotherm plot of Compound (1) HC1 salt Form A.

Figure 17 shows a GVS kinetic plot of Compound (1) HC1 salt Form A.

Figure 18 shows variable temperature XRPD 29 diffractograms of Compound (1) HC1 Form A, displaying a change in form at 159 °C.

Figure 19 shows a XRPD 29 diffractogram of post-variable temperature material compared to XRPD 29 diffractograms of Compound (1) HC1 Forms A and D.

Figure 29 shows an LC-MS spectrum of Compound (1) HC1 salt Form A.

Figure 21 shows and FT-IR spectrum of Compound (1) HC1 salt Form A.

Figure 22 shows an XRPD 29 diffractogram of Compound (1) HC1 salt Form B.

Figure 23 shows a TG/DTA thermogram from 2 °C-299 °C of Compound (1) HC1 salt Form B.

Figure 24A shows a DSC thermogram of the first heating cycle from 29 °C- 299 °C of Compound (1) HC1 salt Form B. Figure 24B shows a DSC thermogram of the first cooling cycle from 299 °C-29 °C of Compound (1) HC1 salt Form B.

Figure 25 shows a DVS isotherm at 25 °C of Compound (1) HC1 salt Form B.

Figure 26 shows an ¹H NMR spectrum of Compound (1) HC1 salt Form B.

Figure 27 shows an FT-IR spectrum of Compound (1) HC1 salt Form B.

Figure 28 shows an XRPD 29 diffractogram of Compound (1) HC1 salt Form D.

Figure 29 shows a TG/DTA thermogram from 2 °C-299 °C of Compound (1)

HC1 salt Form D. Figure 30A shows a DSC thermogram of the first heating cycle from 20 °C- 200 °C of Compound (1) HC1 salt Form D. Figure 30B shows a DSC thermogram of the first cooling cycle from 200 °C-20 °C of Compound (1) HC1 salt Form D.

Figure 31 shows a DVS isotherm at 25 °C of Compound (1) HC1 salt Form D.

Figure 32 shows an ¹H NMR spectrum of Compound (1) HC1 salt Form D.

Figure 33 shows an FT-IR spectrum of Compound (1) HC1 salt Form D.

Figure 34 shows an XRPD 2θ diffractogram of Compound (1) HC1 salt Form G.

Figure 35 shows a TG/DTA thermogram from 2 °C-200 °C of Compound (1) HC1 salt Form G.

Figure 36A shows a DSC thermogram of the first heating cycle from 20 °C- 200 °C of Compound (1) HC1 salt Form G. Figure 36B shows a DSC thermogram of the first cooling cycle from 200 °C-20 °C of Compound (1) HC1 salt Form G.

Figure 37 shows a DVS isotherm at 25 °C of Compound (1) HC1 salt Form G.

Figure 38 shows an ¹H NMR spectrum of Compound (1) HC1 salt Form G.

Figure 39 shows an FT-IR spectrum of Compound (1) HC1 salt Form G.

Figure 40 shows polarized light microscopy (PLM) images of Compound (1) HC1 salt crystalline Forms A, B, D, and G.

Figure 41 shows an XRPD 29 diffractogram of Compound (1) Napsylate salt.

Figure 42 shows an ¹H NMR spectrum of Compound (1) Napsylate salt.

Figure 43 shows a TGA thermogram of Compound (1) Napsylate salt.

Figure 44 shows a DSC thermogram of Compound (1) Napsylate salt.

Figure 45 shows an FT-IR spectrum of Compound (1) Napsylate salt.

Figure 46 shows an XRPD 29 diffractogram of Compound (1) Xinafoate salt Form 1.

Figure 47 shows an ¹H NMR (dmso-d₆, 599.12 MHz) spectrum of Compound (1) Xinafoate salt Form 1.

Figure 48 shows a TGA thermogram of Compound (1) Xinafoate salt Form 1.

Figure 49 shows a DSC thermogram of Compound (1) Xinafoate salt Form 1. Figure 59 shows an FT-IR spectrum of Compound (1) Xinafoate salt Form 1.

Figure 51 shows an XRPD 29 diffractogram of Compound (1) Xinafoate salt

Form 2. Figure 52 shows an ¹H NMR (dmso-de, 500.12 MHz) spectrum of Compound (1)

Xinafoate salt Form 2.

Figure 53 shows a TGA thermogram of Compound (1) Xinafoate salt Form 2.

Figure 54 shows a DSC thermogram of Compound (1) Xinafoate salt Form 2. Figure 55 shows an FT-IR spectrum of Compound (1) Xinafoate salt Form 2.

DETAILED DESCRIPTION

Unless the context requires otherwise, throughout this specification and claims, the words “comprise,” “comprising” and the like are to be construed in an open, inclusive sense; the words “a,” “an,” and the like are to be considered as meaning at least one and are not limited to just one; and the term “about” is to be construed as meaning plus or minus 10%. Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context.

The present invention provides crystalline forms of Zc/V-butyl 4-[[l-[(4- chlorophenyl)methyl]-3-hydroxy-2-oxo-4-pyridyl]methyl] piperazine- 1 -carboxylate (Compound (1)) and certain salts thereof, as shown in Table 1.

Also provided are crystalline forms of l-(4-chlorobenzyl)-3-hydroxy-4-

(piperazin-l-ylmethyl)pyridin-2(lH)-one (Compound (2):

Compound (2) l-(4-chlorobenzyl)-3-hydroxy-4-(piperazin-l-ylmethyl)pyridin-2(lH)-one C17H20CIN3O2 Mol Wt: 333.82

The compounds of the present invention may generally be utilized as the free base or in the form of acid addition salts. In some embodiments, the invention provides a crystalline form having the structure of Compound (1) in the form of its free base. In some embodiments, the invention provides a crystalline form having the structure of Compound (1) in the form of its HC1 salt. In some embodiments, the invention provides a crystalline form having the structure of Compound (1) in the form of its napsylate salt. In some embodiments, the invention provides a crystalline form having the structure of Compound (1) in the form of its xinaphoate salt.

As used herein, the term “free base” refers to Compound (1) devoid, or essentially devoid, of addition of any salt.

In some embodiments, the invention provides a stereoisomerically pure compound having the structure (1) in the form of its pharmaceutically acceptable salt.

A “salt” as is well known in the art includes an organic or inorganic compound in ionic form, capable existing in combination with a counterion. A “pharmaceutically acceptable” salt is a salt formed from an ion that has been approved for human or other animals’ consumption and is generally non-toxic, which possess toxicity profiles within a range that affords utility in pharmaceutical applications.

Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, P -hydroxybutyric, salicylic, galactaric and galacturonic acid.

Non-limiting examples of potential salts of this disclosure include but are not limited to hydrochloride, citrate, glycolate, fumarate, malate, tartrate, mesylate, esylate, cinnamate, isethionate, sulfate, phosphate, diphosphate, nitrate, hydrobromide, hydroiodide, succinate, formate, acetate, di chloroacetate, lactate, p-toluenesulfonate, pamitate, pidolate, pamoate, salicylate, 4-aminosalicylate, benzoate, 4-acetamido benzoate, glutamate, aspartate, glycolate, adipate, alginate, ascorbate, besylate, camphorate, camphorsulfonate, camsylate, caprate, caproate, cyclamate, laurylsulfate, edisylate, gentisate, galactarate, gluceptate, gluconate, glucuronate, oxoglutarate, hippurate, lactobionate, malonate, maleate, mandalate, napsylate, napadisylate, naphthoate, oxalate, oleate, sebacate, stearate, succinate, thiocyanate, undecylenate, and xinafoate.

A “hydrate” is a compound that exists in a composition with water molecules. The composition can include water in stoichiometric quantities, such as a monohydrate or a dihydrate, or can include water in random amounts. As the term is used herein a “hydrate” refers to a solid form, i.e., a compound in water solution, while it may be hydrated, is not a hydrate as the term is used herein.

A “solvate” is a similar composition except that a solvent other that water replaces the water. For example, methanol or ethanol can form an “alcoholate”, which can again be stoichiometric or non-stoichiometric. As the term is used herein a “solvate” refers to a solid form, i.e., a compound in solution in a solvent, while it may be solvated, is not a solvate as the term is used herein.

Hydrates and solvates are held together by weak interactions that are generally broken upon dissolution, similar to salts. When a drug substance in such forms is dissolved in the stomach, intestinal canal or blood of a subject, these hydrate/ solvate forms will generally expose the subject to the same active moiety. It is well known in the art that hydrates and solvates are considered eligible for applications in the same way as salts are.

An “isotope” of a compound of the present invention is a compound having one or more atoms of the compound replaced by an isotope of such atom. For example, isotopes include compounds with deuterium in place of one or more hydrogen atoms of Compound (1). Isotopic substitutions which may be made in the formation of isotopes of the present invention include non-radioactive (stable) atoms such as deuterium and carbon 13, as well as radioactive (unstable) atoms such as tritium, carbon 14, iodine 123, iodine 125, and the like.

In one embodiment is provided a crystalline form of an HC1 salt of a compound having the following structure (1):

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is anhydrous.

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is Form B. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 16.8°, 18.6°, 15.2°, 9.0°, and 20.3°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 4.8°, 22.6°, 18.8°, 19.9°, and 29.7°. In another embodiment, crystalline Form B of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 16.8°, 18.6°, 15.2°, 9.9°, and 29.3°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 4.8°, 22.6°, 18.8°, 19.9°, and 29.7°. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 22. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting. In another embodiment, crystalline Form B of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C. In a further embodiment, the melting isotherm has an onset at about 189 °C and a peak at about 193 °C. In another embodiment, the DTA thermogram has a melting isotherm with an onset at about 193 °C and a peak at about 195 °C. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a TG/DTA thermogram substantially as depicted in Figure 23.

In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting. In one embodiment, the DSC thermogram shows a melting endotherm with onset at about 196 °C and a peak at about 198 °C. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 24A or Figure 24B.

In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 26.

In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 25.

In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2344 cm^-1, 2963 cm^-1, 3062 cm^-1, 3179cm^-1. In one embodiment, crystalline Form B of the HC1 salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 27.

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is Form D. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 4.3°, 9.0°, 16.9°, 18.1°, and 22.6°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 14.7°, 15.2°, 16.2°, 18.6°, and 19.3°. In another embodiment, crystalline Form D of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 4.3°, 9.0°, 16.9°, 18.1°, and 22.6°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 14.7°, 15.2°, 16.2°, 18.6°, and 19.3°. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 28.

In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C. In a further embodiment, the DTA thermogram shows a melting isotherm with an onset at about 191 °C and a peak at about 194 °C. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a TG/DTA thermogram substantially as depicted in Figure 29.

In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting. In one embodiment, the DSC thermogram shows a melting endotherm with onset at about 192 °C and a peak at about 196 °C. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 30A or Figure 30B.

In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 32.

In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 0.8% at 90% relative humidity during first sorption. In another embodiment, crystalline Form D of the HC1 salt of Compound (1) has a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 31. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) is class II hygroscopic.

In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2349 cm^-1, 2933 cm^-1, 3062cm^-1, and 3177 cm^-1. In one embodiment, crystalline Form D of the HC1 salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 33. In one embodiment, the crystalline form of the HC1 salt of Compound (1) is Form G. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 4.7°, 9.5°, 13.2°, 16.7°, and 19.3°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 11.2°, 19.1°, 19.5°, 21.2°, and 24.5°. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 4.7°, 9.5°, 13.2°, 16.7°, and 19.3°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 9.5°): 11.2°, 19.1°, 19.5°, 21.2°, and 24.5°. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 34.

In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting. In another embodiment, crystalline Form G of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C. In a further embodiment, the DTA thermogram shows a melting isotherm with an onset at about 191 °C and a peak at about 195 °C. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a TG/DTA thermogram substantially as depicted in Figure 35.

In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting. In one embodiment, the DSC thermogram shows a melting endotherm at about 192 °C and a peak at about 196 °C. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 36A or Figure 36B.

In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a proton nuclear magnetic resonance (^rH NMR) spectrum substantially as depicted in Figure 38.

In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 9.8% at 99% relative humidity during first sorption. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 37. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) is class II hygroscopic. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2349 cm^-1, 2933 cm^-1, 3062cm^-1, and 3177 cm^-1. In one embodiment, crystalline Form G of the HC1 salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 39.

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is a monohydrate. In a further embodiment, the crystalline form of the HC1 salt of Compound (1) is Form A.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 15.1°, 17.4°, 19.8°, 20.0°, and 20.6°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 15.9°, 15.3°, 16.2°, 17.3°, and 26.9°. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 15.1°, 17.4°, 19.8°, 29.9°, and 29.6°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 15.9°, 15.3°, 16.2°, 17.3°, and 26.9°. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 12.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 3.4 % from about 29 °C to about 89 °C. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 53 °C. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a TG/DTA thermogram substantially as depicted in Figure 13.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a differential scanning calorimetry (DSC) thermogram having an endothermic peak with onset at about 79 °C and a peak at about 116 °C. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 14A, Figure 14B, or Figure 14C.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a proton nuclear magnetic resonance NMR) spectrum substantially as depicted in Figure 15 A. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a heteronuclear single quantum correlation (HSQC) spectrum substantially as depicted in Figure 15B.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a gravimetric vapor sorption (GVS) isotherm plot substantially as depicted in Figure 16. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a GVS kinetic plot substantially as depicted in Figure 17.

In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 764 cm^-1, 1657 cm^-1, 2977 cm^-1, 3202 cm^-1, and 3483 cm^-1. In one embodiment, crystalline Form A of the HC1 salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 21.

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is a chloroform solvate. In a further embodiment, the crystalline form of the HC1 salt of Compound (1) is Form F.

In one embodiment, crystalline Form F of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 17.6 % from about 20 °C to about 150 °C. In one embodiment, crystalline Form F of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 191 °C and a peak at about 195 °C.

In one embodiment, the crystalline form of the HC1 salt of Compound (1) is a anisole solvate. In a further embodiment, the crystalline form of the HC1 salt of Compound (1) is Form H.

In one embodiment, crystalline Form H of the HC1 salt of Compound (1) has a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 10.0 % from about 20 °C to about 130 °C. In one embodiment, crystalline Form H of the HC1 salt of Compound (1) has a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 64 °C.

In one embodiment is provided a crystalline form of the free base of a compound having the following structure (1):

In one embodiment, the crystalline form of the free base of Compound (1) is Form E. In one embodiment, crystalline Form E of the free base of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 13.5°, 16.7°, 17.9°, 18.5°, and 19.8°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 12.3°, 18.4°, 19.5°, 21.2°, and 22.7°. In one embodiment, crystalline Form E of the free base of Compound (1) has an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 13.5°, 16.7°, 17.9°, 18.5°, and 19.8°. In a further embodiment, the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 12.3°, 18.4°, 19.5°, 21.2°, and 22.7°. In one embodiment, crystalline Form E of the free base of Compound (1) has an XRPD spectrum substantially as depicted in Figure 2.

In one embodiment, crystalline Form E of the free base of Compound (1) has a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting. In a further embodiment, crystalline Form E of the free base of Compound (1) has a melting endotherm with a peak at about 188°C. In one embodiment, crystalline Form E of the free base of Compound (1) has a DSC thermogram substantially as depicted in Figure 5.

In one embodiment, crystalline Form E of the free base of Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 4.

In one embodiment, crystalline Form E of the free base of Compound (1) has a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 9.2% at 99% relative humidity during first sorption. In one embodiment, crystalline Form E of the free base of Compound (1) has a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 6. In one embodiment, crystalline Form E of the free base of Compound (1) is hygroscopic. In one embodiment, crystalline Form E of the free base of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 1065 cm^-1, 2816 cm^-1, 2848 cm^-1, 2961 cm^-1, 2976 cm^-1, and 3085 cm^-1. In one embodiment, crystalline Form E of the free base of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 7.

In one embodiment, single crystal Form E of the free base of Compound (1) has a primitive orthorhombic lattice Bravais type. In a further embodiment, Form E has a space group of Pbca. In a further embodiment, the primitive orthorhombic lattice comprises vectors wherein a is about 10.4186 A, a is about 90°, b is about 11.6775 A, p is about 90°, c is about 35.442 A, and y is about 90°.

In one embodiment is provided a crystalline form of a napsylate salt of a compound having the following structure (1):

In one embodiment, the crystalline form of the napsylate salt of Compound (1) is Napsylate Form 1. In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 41.

In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of Compound (1) has a TGA thermogram substantially as depicted in Figure 43.

In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 42.

In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 44. In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of

Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 622 cm^-1, 648 cm^-1, 867 cm^-1, 1394 cm^-1, 1663 cm^-1, 2979 cm^-1. In one embodiment, crystalline Napsylate Form 1 of the napsyalte salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 45.

In one embodiment is provided a crystalline form of a xinafoate salt of a compound having the following structure (1):

In one embodiment, the crystalline form of the xinafoate salt of Compound (1) is Xinafoate Form 1. In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 46.

In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of Compound (1) has a TGA thermogram substantially as depicted in Figure 48.

In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of

Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 47.

In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 49.

In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 532 cm^-1, 586 cm^-1, 659 cm^-1, 1494 cm^-1, 1657 cm^-1, 2976 cm^-1. In one embodiment, crystalline Xinafoate Form 1 of the xinafoate salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 50. In one embodiment, the crystalline form of the xinafoate salt of Compound (1) is Xinafoate Form 2. In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has an XRPD spectrum substantially as depicted in Figure 51.

In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has a TGA thermogram substantially as depicted in Figure 53.

In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 52.

In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has a DSC thermogram substantially as depicted in Figure 54.

In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 685 cm^-1, 876 cm^-1, 925 cm^-1, 1493 cm^-1, 1741 cm^-1, 2983 cm^-1. In one embodiment, crystalline Xinafoate Form 2 of the xinafoate salt of Compound (1) has an FT-IR spectrogram substantially as depicted in Figure 55.

In one embodiment is provided a crystalline form of a compound having the following structure (2):

In one embodiment, the crystalline form of Compound (2) has an XRPD spectrum substantially as depicted in Figure 10.

In certain embodiments, the invention provides a pharmaceutical composition comprising Compound (1), or a pharmaceutically acceptable salt, hydrate, solvate, or isotope thereof, or a crystalline form thereof; and a pharmaceutically acceptable carrier. In a further embodiment, the present pharmaceutical composition comprises the free base form of Compound (1). In further embodiments, the present pharmaceutical composition comprises the HC1, calcium, napsylate, xinafoate, or 3-hydroxy-2-napthoate salt of Compound (1).

Pharmaceutically acceptable carrier can be those familiar to persons skilled in the art. The particular carrier employed in these pharmaceutical compositions may vary depending upon the type of administration desired (e.g. intravenous, oral, topical, suppository, or parenteral). For compositions formulated as liquid solutions, acceptable carriers include saline and sterile water, and may optionally include antioxidants, buffers, bacteriostats and other common additives. The compositions can also be formulated as pills, capsules, granules, or tablets which contain, in addition to Compound (1), excipients such as diluents, binders, and lubricants. One skilled in this art may further formulate Compound (1) in an appropriate manner, and in accordance with accepted practices, such as those disclosed in Remington: The Science and Practice of Pharmacy, 22^nd Edition, Allen, Lloyd V., Jr. Ed. (2012) (incorporated herein by reference).

For example, the compounds or crystalline forms of the invention will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which can be in the form of an ampoule, capsule, sachet, paper, or other container. When the compounds or crystalline forms of the invention is mixed with a carrier, or when the carrier serves as a diluent, it can be solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The compounds or crystalline forms of the invention can be adsorbed on a granular solid carrier, for example contained in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxy ethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrier can include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.

In certain embodiments, the invention provides a method for inhibiting an HIF-1α prolyl hydroxylase, comprising contacting the HIF-1α prolyl hydroxylase with an effective amount of Compound (1), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, or a crystalline form thereof, or a pharmaceutical composition comprising the same. In further embodiments, the present method comprises administering the free base form of Compound (1). In other embodiments, the present method comprises administering the HC1, calcium, napsylate, xinafoate, or 3-hydroxy-2-napthoate salt of Compound (1).

The expression “effective amount”, when used to describe use of a compound or composition of the invention, refers to the amount of a compound or composition of the invention that is effective to inhibit a HIF-1α prolyl hydroxylase. When applied in a subject and the HIF-1α prolyl hydroxylase is implicated in a medical disorder or condition, such binding occurs to an extent sufficient to produce a beneficial therapeutic effect on the subject. Similarly, as used herein, an “effective amount” of a compound or composition of the invention refers to an amount of the compound or composition that alleviates, in whole or in part, symptoms associated with the disorder or condition, or halts or slows further progression or worsening of those symptoms, or prevents or provides prophylaxis for the disorder or condition, in particular, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result by acting as an inhibitor of HIF-1α prolyl hydroxylase. A therapeutically effective amount is also one in which any toxic or detrimental effects of compounds or compositions of the invention are outweighed by the therapeutically beneficial effects.

In more specific embodiments, the invention provides a method for treating a disease or condition for which inhibition of HIF-1α prolyl hydroxylase is beneficial, comprising administering to a subject in need thereof an effective amount of Compound (1), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, or a crystalline form thereof, or a pharmaceutical composition comprising the same. In further embodiments, the present method comprises administering the free base form of Compound (1). In other embodiments, the present method comprises administering the HC1, calcium, napsylate, xinafoate, or 3-hydroxy-2- napthoate salt of Compound (1).

“Treating” or “treatment” within the meaning herein refers to an alleviation of symptoms associated with a medical disorder or disease, or inhibition of further progression or worsening of those symptoms, or prevention or prophylaxis of the disease or disorder.

As used herein, “subject” means warm-blood animals, including, for example, humans; non-human primates, e.g. apes and monkeys; cattle; horses; sheep; and goats.

“Administering” or “administration” can be conducted through any route of administration which effectively transports the active compound of the invention which inhibits the NK-3 receptor to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal or parenteral, e.g., rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment, the oral route being preferred. The administration can be local or systemic, preferably, through the pharmaceutical compositions as discussed above. As used herein, systemic administration includes, for example, oral and parenteral methods of administration, subcutaneous, intramuscular, intracranial, intraorbital, ophthalmic, intraventricular, intracapsular, intraarticular, intraspinal, intracisternal, intraperitoneal, intranasal, aerosol, intravenous, intradermal, inhalational, transdermal, transmucosal, and rectal administration.

In more specific embodiments, the invention provides a method for treating an inflammatory disease, comprising administering to a subject in need thereof an effective amount of Compound (1), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, or a crystalline form thereof, or a pharmaceutical composition comprising the same. In further embodiments, the present method comprises administering the free base form of Compound (1). In other embodiments, the present method comprises administering the HC1, calcium, napsylate, xinafoate, or 3-hydroxy-2-napthoate salt of Compound (1). In some embodiments, the inflammatory disease is an inflammatory bowel disease. In further embodiments, the inflammatory bowel disease is ulcerative colitis. In another embodiment, the inflammatory bowel disease is Crohn’s disease.

EXAMPLES

In order that this invention may be more fully understood, the following examples are set forth. These examples are for the purpose of illustrating embodiments of this invention, and are not to be construed as limiting the scope of the invention in any way. The reactants used in the examples below may be obtained either as described herein, or if not described herein, are themselves either commercially available or may be prepared from commercially available materials by methods known in the art.

Polymorph screen of Compound (1) HC1 in 26 solvent systems under varying conditions, identified six novel XRPD patterns (identified as Forms A, B, D, E, F and G).

The forms identified from the Polymorph Screen are characterized below: Form A - a crystalline powder with small crystals with plate- like morphology and slight hygroscopicity, identified as a stoichiometric hydrate of Compound (1) HC1.

Form B - an anhydrous / non solvated form, recovered from temperature cycling in pure organic systems, (primary alcohols, acetone, methylisopropyl ketone, isopropyl acetate). A free-flowing crystalline powder with large, rod shaped crystals. Very low hygroscopicity and stable even under prolonged exposure to elevated temperatures/high humidity. Some conversion to Compound (1) HC1 Form A was observed when exposed to solvents with higher water activity.

Form D - an anhydrous / non-solvated form, recovered from temperature cycling in THF, tert-butylmethyl ether and 1,4-di oxane. A free- flowing, crystalline powder of small crystals with irregular morphology. Slightly hygroscopic and stable at ambient though slowly converts to Form B.

Form E - recovered from temperature cycling in deionized water, identified as Compound (1) Free Base, indicating the HC1 salt dissociates to its free base when stored for a prolonged period under aqueous conditions.

Form F - recovered from temperature cycling in chloroform identified as a chloroform solvate. Unstable form that converts to Form B upon drying.

Form G - a non-solvated / anhydrous form, recovered from temperature cycling in methyl ethyl ketone, ethyl and propyl acetate and 2-m ethyl THF. An agglomerated solid with acicular rod-shaped crystals. Slightly hygroscopic and stable at ambient conditions, while increased temperatures/high humidity initiate conversion to a mixture of Forms D and B.

Form H - a hemi-anisole solvate, recovered from temperature cycling in anisole. An unstable form that converts to Form D (through Form G) upon drying.

The relationship between the various crystal forms is shown in FIG 1.

Form A is very stable, prepared utilizing water-based crystallization, and preferred in systems with high water activity, in particular in API manufacturing and wet granulation processes where water may be present.

Stable Forms B, D and G were scaled-up using linear procedures from the primary polymorph screen, recovered with yields of 78.8%, 68.1% and 81.0%, respectively.

Form B was found to be stable across a wide range of conditions tested, including retaining structural integrity under high temperatures/RH, against contamination with other forms (while in a slurry seeded with Forms D and G). It also showed stability in water activity below 0.3 (methanol), with indications of conversion into Form A above that threshold. Forms D and G were found to be stable only under ambient conditions and when no other forms were present. Based on these observations, Form B is the most stable anhydrous, non-solvated form of Compound (1) HC1 and Form A is preferred in systems with high water activity.

EXAMPLE 1

Analytical Methods

A. X-ray Powder Diffraction (XRPD)

XRPD analysis was carried out on a PANalytical X’pert pro fitted with PIXcel detector, scanning the samples between 3 and 35° 29. The material was gently ground to release any agglomerates and loaded onto a multi-well plate with Kapton or Mylar polymer film to support the sample. The multi-well plate was then placed into the diffractometer and analyzed using Cu K radiation (al = 1.54060 A; a2 = 1.54443 A; P = 1.39225 A; al : a2 ratio = 0.5) running in transmission mode (step size 0.0130° 29) using 40 kV / 40 mA generator settings.

B. Polarized Light Microscopy (PLM)

Crystallinity (birefringence) was determined using an Olympus BX50 polarizing microscope, equipped with a Motic camera and image capture software (Motic Images Plus 2.0). Images were typically recorded using the 20x objective.

C. Thermogravimetric Analysis (TGA) and Differential Thermal Analysis (DTA)

Approximately, 5mg of material was weighed into an open aluminum pan and loaded into a simultaneous thermogravimetric/differential thermal analyzer (TG/DTA) and held at room temperature. The sample was then heated at a rate of 10 °C/minute from 20 °C to 300 °C (unless otherwise stated) during which time the change in sample weight was recorded along with any differential thermal analysis (DTA) events. Nitrogen was used as purge gas, at a flow rate of 300 cm³/minute. D. Differential Scanning Calorimetry (DSC)

Approximately, 5mg of material was weighed into an aluminum DSC pan and sealed non- hermetically with a pierced aluminum lid. The sample pan was then loaded into a Seiko DSC6200 (equipped with a cooler) cooled and held at 20 °C. Once a stable heat-flow response was obtained, the sample and reference were heated to 300 °C (unless otherwise stated) at scan rate of 10 °C/minute and the resulting heat flow response monitored. Nitrogen was used as the purge gas, at a flow rate of 50 cm3/minute.

E. Infrared Spectroscopy (IR)

Infrared spectroscopy was carried out on a Bruker ALPHA P spectrometer.

Sufficient material was placed onto the center of the plate of the spectrometer and the spectra were obtained using the following parameters:

Resolution: 4 cm'¹

Background Scan Time: 16 scans

Sample Scan Time: 16 scans

Data Collection: 4000 to 400 cm'¹

Result Spectrum: Transmittance

Software: OPUS version 6

F. Nuclear Magnetic Resonance (NMR)

NMR experiments were performed on a Bruker AVIHHD spectrometer equipped with a DCH cryoprobe operating at 500.12 MHz for protons. Experiments were performed in deuterated DMSO and each sample was prepared to ca. 10 mM concentration.

G. Dynamic Vapor Sorption (DVS)

Approximately, 10-20 mg of sample was placed into a mesh vapor sorption balance pan and loaded into a DVS Advantage dynamic vapor sorption balance by Surface Measurement Systems. The sample was subjected to a ramping profile from 40 - 90% relative humidity (RH) at 10% increments, maintaining the sample at each step until a stable weight had been achieved (dm/dt 0.004%, minimum step length 30 minutes, maximum step length 500 minutes) at 25 °C. After completion of the sorption cycle, the sample was dried using the same procedure to 0% RH and then a second sorption cycle back to 40% RH. Two cycles were performed. The weight change during the sorption/desorption cycles were plotted, allowing for the hygroscopic nature of the sample to be determined. Where applicable, non-ambient analysis was conducted under temperature stated, using settings above. XRPD analysis was then carried out on any solid retained.

H. Gravimetric Vapor Sorption (GVS)

Approximately 10-20 mg of sample was placed into a mesh vapor sorption balance pan and loaded into an IGASorp Moisture Sorption Analyzer balance (Hiden Analytical). The sample was subjected to a ramping profile from 40 - 90% relative humidity (RH) at 10% increments, maintaining the sample at each step until a stable weight had been achieved (98% step completion, minimum step length 30 minutes, maximum step length 60 minutes) at 25 °C. After completion of the sorption cycle, the sample was dried using the same procedure to 0% RH, and finally taken back to the starting point of 40% RH. Two cycles were performed. The weight change during the sorption/desorption cycles were plotted, allowing for the hygroscopic nature of the sample to be determined. XRPD analysis was then carried out on any solid retained.

I. Variable Temperature X-ray Powder Diffraction (VT-XRPD)

VT-XRPD analysis was carried out on a Philips X’Pert Pro Multipurpose diffractometer equipped with a temperature chamber. The samples were scanned between 4 and 35.99 °29 using Cu K radiation (al = 1.54060 A; a2 = 1.54443 A; P = 1.39225 A; al : a2 ratio = 0.5) running in Bragg-Brentano geometry (step size 0.008 °29) using 40 kV / 40 mA generator settings. Measurements were performed using the following temperature profiles, at a heating rate used of 10 °C/minute.

Profile 1 : 30 °C, 1 minute hold, scan

Heat to 150 °C, 5 minute hold, scan; 10 minute hold, scan Cool to 30 °C, 1 minute hold, scan.

Profile 2: 30 °C, 5 minutes hold, scan

Heat to 60 °C, 5 minutes hold, scan

Heat to 70 °C, 5 minutes hold, scan Heat to 80 °C, 5 minutes hold, scan Heat to 90 °C, 5 minutes hold, scan Heat to 100 °C, 5 minutes hold, scan Heat to 110 °C, 5 minutes hold, scan Heat to 120 °C, 5 minutes hold, scan Heat to 130 °C, 5 minutes hold, scan

Heat to 160 °C, 5 minutes hold, scan Cool to 30 °C, 5 minutes hold, scan

J. High Performance Liquid Chromatography-Ultraviolet Detection (HPLC-UV)

Instrument: Dionex Ultimate 3000

Column: Waters Sunfure C18 5pm 150 mm * 4.6 mm

Column Temperature: 40 °C

UV wavelength: 240 nm

Injection Volume: 10 pL

Flow Rate: 1.0 mL/min

Mobile Phase A: 20 mM KH2PO4 pH 6.0

Mobile Phase B: 20 mM KH₂PO₄:ACN (20:80 % v/v)

Gradient Program: Time (minutes) Solvent B [%]

0.0 40.0

20.0 100.0

25.0 100.0

25.1 40.0

35.0 40.0

K. High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS)

Column: ACE EXCEL3 C18, 3.0pm, 75 x 4.6mm.

Mobile Phase A: 0.1% Formic Acid in Deionised water

Mobile Phase B: 0.1% Formic Acid in Acetonitrile

Diluent: Acetonitrile: Water (50:50 % v/v)

Flow Rate: 1.0 mL/min Runtime: 20 minutes

Column Temperature: 30 °C

Injection Volume: 10 pL

Needle Wash Acetonitrile

PDA Range : 210 nm

Gradient Program: Time (minutes) Solvent B [%]

0 5

12.00 95

15.00 95

15.10 5

20.00 5

Both +ve and -ve ESI used.

Tune method:

Capillary temp: 200 °C

Sheath Gas Flow: 20.0

S ource Voltage : 4.50 k V

Source Current: 80.00 uA

Capillary Voltage: 8.0 V

Tube Lens Offset: 40.00 V

L. High Performance Liquid Chromatography-Charged Aerosol Detection (HPLC-CAD)

Instrument: Dionex Ultimate 3000

Column: ACE 3 Hilic-N, 100x3 mm, 3 pm

Column Temperature: 30 °C

Autosampler Temperature: 30 °C

Injection Volume: 15 pL

Flow Rate: 0.6 mL/min

Mobile Phase A: 100 mM Ammonium Formate Buffer (pH 3.65) :

Acetonitrile (10:90 v/v)

Mobile Phase B: 100 mM Ammonium Formate Buffer (pH 3.65) :

Acetonitrile (90: 10 v/v) Gradient Program: Time (minutes) Solvent B [%]

0 0.5

5.0 0.5

35.0 99.5

40.0 99.5

40.10 0.5

50.0 0.5

CAD Parameters

Gain: 100 pA

Offset: 0% Nebuliser Temperature: 30 °C

EXAMPLE 2

Characterization of Compound (1) Free Base

A. X-ray Powder Diffraction (XRPD) of Compound (1) Free Base

FIG 2 shows the XRPD 29 Diffractogram of Compound (1) Free Base; the corresponding peak list is presented in Table 2. The XRPD diffractogram of Compound (1) Free Base is consistent with the diffractogram pattern of Compound (1) HC1 Form E (see FIG 3).

Table 2: XRPD peak list of Compound (1) Free Base

No. Pos. [20] Area [cts* 20] d-spacing [A] Height [cts] Rel. Int. [%]

1 4.92660 65.74 17.93747 1041.95 19.91

2 12.25660 114.17 7.22155 1507.98 28.82

3 13.45380 204.28 6.58149 2312.69 44.19

4 14.94480 101.14 5.92807 1145.00 21.88

5 16.70960 528.29 5.30573 5233.26 100.00

6 17.87180 310.46 4.96324 3075.40 58.77

7 18.36240 94.02 4.83171 1241.84 23.73 8 18.53020 456.56 4.78835 4522.62 86.42

9 19.00810 62.26 4.66903 616.79 11.79

10 19.47490 170.72 4.55815 1503.25 28.72

11 19.84010 216.86 4.47507 1909.49 36.49

12 21.18790 176.48 4.19335 1398.58 26.72

13 22.42820 69.50 3.96419 550.75 10.52

14 22.67490 129.73 3.92160 1285.06 24.56

15 23.07420 122.12 3.85463 967.74 18.49

16 23.76800 161.40 3.74366 1162.80 22.22

17 27.12040 84.89 3.28804 560.64 10.71

18 27.39640 42.92 3.25554 566.87 10.83

19 28.26000 53.89 3.15798 305.07 5.83

20 29.19610 45.08 3.05883 357.28 6.83

B.

Compound (1) Free Base

FIG 4 shows the ^XH NMR spectrum of Compound (1) Free Base.

C. DSC thermogram of Compound (1) Free Base

FIG 5 shows the DSC thermogram of Compound (1) Free Base. D. DVS Isotherm of Compound (1) Free Base

FIG 6 shows the DVS isotherm of Compound (1) Free Base.

E. FT-IR of Compound (1) Free Base

FIG 7 shows the FT-IR spectrum of Compound (1) Free Base and the corresponding peak listing is presented below in Table 3. Table 3: Compound (1) Free Base FT-IR Peaks

Wavenumber Absolute intensity

1689.65 0.75

1654.34 0.87

1597.59 0.77 Wavenumber Absolute intensity

1489.73 0.88

1455.02 0.83

1444.03 0.87

1405.88 0.79

1364.02 0.84

1304.53 0.88

1290.69 0.87

1262.91 0.86

1248.04 0.77

1216.50 0.80

1190.58 0.78

1161.14 0.78

1118.76 0.78

1092.02 0.76

1064.92 0.89

1016.92 0.88

1007.55 0.86

994.24 0.78

966.59 0.89

867.05 0.86

846.96 0.85

831.29 0.87

809.77 0.86

784.08 0.84

761.84 0.77

729.76 0.88

557.05 0.81

543.93 0.89

515.88 0.81

479.31 0.89

449.20 0.89

410.98 0.85

2961.35 0.93

2847.78 0.94 Wavenumber Absolute intensity

2815.79 0.94

3085.10 0.96

2975.52 0.93

EXAMPLE 3

Single Crystal Analysis of Compound (1) Freebase

A. Single Crystal Growth

Compound (1) Freebase (10-20 mg) was dissolved in dichloromethane (500 pL) to obtain a saturated solution. Hexane (2 mL) was added as antisolvent and then left at ambient temperature. After 2 weeks, suitable crystals had formed.

B. Single Crystal X-ray Analysis (SXRD)

A suitable crystal of Compound (1) Free Base Form 1 was selected and mounted in a loop using paratone oil. Data were collected using a Bruker D8Venture diffractometer equipped with a Photon III detector operating in shutterless mode at 100(2) K with Cu-Ka radiation (1.54178 A). The structure was solved in the Olex2 software package with the ShelXT (intrinsic phasing) structure solution program and refined with the ShelXL refinement package using Least Squares minimisation. Data were collected, solved and refined in the orthorhombic space-group Pbca.

All non-hydrogen atoms were located in the Fourier map and their positions refined prior to describing the thermal movement of all non-hydrogen atoms anisotropically.

Within the asymmetric unit, one complete Compound (1) formula unit was refined. All hydrogen atoms were placed in calculated positions using a riding model with fixed Uiso at 1.2 times for all CH and CHi groups, and 1.5 times for all CH3 and OH groups. See FIG 8 for structure.

The highest residual Fourier peak was found to be 0.43 e.A'³ , approx. 0.86 A from Cl( 1 ) and the deepest Fourier hole was found to be -0.50 e.A'³ , approx. 0.64 A from Cl( 1 ). C. Crystal Data & Structural Features

The unit cell dimensions are:

Orthorhombic, space group Pbca a = 10.4186(9) A a = 90° b = 11.6775(8) A = 90° c = 35.442(2) A y = 90°

Volume = 4312.0(6) A3

Z = 8, Z' = 1

V= 4312.0(6) A³

T= 100(2) K p(CuKa) = 1.850 mm"¹

66439 reflections were collected (4.986° < 20 < 144.634°), 4267 unique (/Cu = 0.0835, Rsigma = 0.0279) which were used in all calculations.

The final R\ was 0.0391 (>2c(I)) and wRi was 0.0982 (all data).

Goodness of fit = 1.027

Rint = 8.35 %

The asymmetric unit contained one complete Compound (1) Free Base Form 1 formula unit and did not contain any solvent accessible voids or regions of electron density that could be accounted for as unassigned solvent when viewed along unit cell axes a, b and c.

Packing within the structure was shown to be efficient with a density of 1.337 g.cm"³.

Within the packing, a hydrogen bonding network was noted between N(2) • • H(3) with a length of 1.9969(13) A and an N(2) — H(3) — 0(3) angle of 151.51(8)°, propagated along the Z>-axis. All other interactions were due to van der Waals forces.

A simulated XRPD diffractogram was calculated which was consistent with the experimental diffractogram (FIG 9). EXAMPLE 4

Characterization of Compound (2)

Compound (2)

C17H20CIN3O2 Mol Wt: 333.82

FIG 10 shows the XRPD 29 diffractogram of Compound (2) which was clearly distinct from the other patterns identified for Compound (1).

TG analysis (FIG 11) showed an initial mass loss of 4.97% from the onset of heating to ca. 62 °C related to the loss of surface moisture (4.97%, ca. 1.26 equivalents of water). The material remained stable to 155 °C where a 2.31 % mass loss was observed during melting transition, after which thermal degradation was observed. DT analysis showed a broad, shallow endothermic event corresponding to the loss of surface moisture (onset ca. 35 °C), followed by a shallow melting endotherm (onset at ca. 187 °C) and a peak at ca. 192 °C, followed by material degradation. The melting transition is slightly lower than that of Compound (1) HC1 (Compound (1) HC1 melting point onset at 190°C / peak at 194 °C, Compound (2) melting point onset at 187 °C / peak at 192 °C).

The

NMR spectrum was consistent with the compound structure; the singlet corresponding to the BOC group (1.4 ppm) was absent. EXAMPLE 5

Characterization of Compound (1) HC1 salt. Form A

A. XRPD of Compound (1) HC1 salt Form A

FIG 12 shows the XRPD 29 Diffractogram of Compound (1) HC1 salt Form A; the corresponding peak list in presented in Table 4. Polarized Light Microscopy showed small birefringent crystals (ca. 20- 100pm) with plate-like morphology.

Table 4: XRPD peak list of Compound (1) HC1 salt, Form A

No. Pos. [20] Area [cts* 20] d-spacing [A] Height [cts] Rel. Int. [%]

1 5.91200 50.07 14.94968 793.52 21.30

2 15.00140 93.68 5.90583 1856.06 49.81

3 15.11510 259.59 5.86166 2938.83 78.87

4 15.34350 116.84 5.77491 1322.78 35.50

5 15.60160 89.41 5.67997 885.73 23.77

6 15.93380 44.34 5.56228 702.71 18.86

7 16.23020 161.42 5.46135 1599.02 42.91

8 17.27050 114.45 5.13467 1511.64 40.57

9 17.37160 166.49 5.10501 2198.95 59.01

10 19.77420 417.34 4.48983 3674.85 98.62

11 20.04290 470.19 4.43025 3726.15 100.00

12 20.57610 241.80 4.31663 2129.11 57.14

13 20.76170 54.15 4.27845 715.23 19.19

14 21.58470 97.17 4.11715 700.03 18.79

15 23.15250 59.46 3.84178 673.16 18.07

16 25.75900 120.64 3.45864 1195.02 32.07

17 25.92860 178.40 3.43641 1570.90 42.16

18 26.53970 148.49 3.35866 840.55 22.56

19 26.85890 102.20 3.31945 809.91 21.74

20 30.12340 136.45 2.96675 831.79 22.32 B. Thermogravimetric/differential thermal analyzer (TG/DTA) of Compound (1) HC1 salt,

Form A

FIG 13 shows the TG/DTA Thermogram of Compound (1) HC1 Form A. Thermogravimetric analysis showed an initial mass loss (from onset to ca. 80 °C) of 3.4% related to loss of water (ca. 0.9 equiv). A loss of ca. 1 equivalent of water indicated Compound (1) HC1 form A is a monohydrate. A second mass loss of 40.9% was observed (with an onset at ca. 160 °C) related to the material melt and subsequent decomposition.

Differential thermal analysis showed a shallow, broad endothermic event (onset at ca. 53 °C) related to the loss of water. An initially sharp then broadening melt was recorded from an onset of ca. 190 °C and a peak at ca. 194 °C (potentially related to material degradation).

C. Differential Scanning Calorimetry (DSC) of Compound (1) HC1 salt, Form A

FIG 14 shows DSC Thermograms of Compound (1) HC1 Form A, (i) first heating cycle, 20-150 °C; (ii) first cooling cycle, 150-20 °C; and (iii) second heating cycle, 20-300 °C. DSC analysis showed a broad endothermic event with onset at ca. 70 °C related to the loss of water during the first heating cycle (up to 150 °C), as well as a sharp melt endothermic event with an onset at 191 °C and peak at 194 °C during the second heating cycle (up to 300 °C). The melting point was consistent with TG/DTA.No endothermic events were detected during the second heat cycle in the solvent loss region (up to ca. 150 °C), suggesting that originally monohydrated material dehydrated during the first heat phase. No significant thermal events were observed during cooling or the second heating cycle.

D. NMR of Compound (1) HC1 salt

FIG 15 shows (i) the ¹H-NMR and (ii) Heteronuclear Single Quantum Correlation (HSQC) spectra of Compound (1) HC1.

E. Gravimetric Vapor Sorption (GVS) of Compound (1) HC1 salt, Form A

FIG 16 shows a GVS isotherm plot, and FIG 17 shows a GVS kinetic plot of Compound (1) HC1 Form A. GVS analysis showed the material to be slightly hygroscopic, with a maximum mass increase of 0.3% at 90% RH during the second sorption cycle. The difference in mass uptake in the first sorption cycle (ca. 0.17%) and in the second sorption cycle (ca. 0.3%) was potentially due to the monohydrate nature and thus, having a limited moisture uptake capability. The increased noise level during the second cycle may have also been a contributing factor. During the first desorption cycle, the material potentially converted to the anhydrous form which allowed for an increase moisture uptake during the second sorption cycle. XRPD analysis post-GVS showed no change in form after exposure to the GVS humidity conditions.

F. VT-XRPD of Compound (1) HC1 salt

FIG 18 shows Variable Temperature XRPD 20 Diffractograms of Compound (1)

HC1 Form A, displaying a change in form at 150 °C, potentially due to dehydration. Post VT-

XRPD Transmission XRPD diffractogram resembled Form D (FIG 19), suggesting this polymorph might be the anhydrous form. TG/DT analysis on the post-VT XRPD material confirmed the material was anhydrous, as no mass loss was observed until thermal degradation.

G. HPLC of Compound (1) HC1 salt, Form A

HPLC analysis recovered a purity of 99.6% by area HPLC Integration; see Table

5 for integration results. Table 5: HPLC Integration results

Relative Relative

Retention Retention Area Height Area

No. Peak Name Time Time mAU*min mAU %

Injection 1

1 3.410 0.26 0.197 0.429 0.20

2 12.590 0.96 0.114 0.385 0.11

3 Compound (1) 13.050 1.00 100.168 824.188 99.63

4 16.453 1.26 0.061 0.390 0.06

Injection 2

1 3.440 0.26 0.206 0.478 0.02

2 12.610 0.96 0.130 0.428 0.13

3 Compound (1) 13.070 1.00 101.475 838.322 99.62

4 16.470 1.26 0.053 0.391 0.05 High Performance Liquid Chromatography-Charged Aerosol Detection (HPLC- C D) analysis confirmed Compound (1) to be a mono-HCl salt.

H. LC-MS of Compound (1) HC1 salt Form A

FIG 20 shows the LC-MS spectrum of Compound (1) HC1 salt, demonstrating an observed a mass of 434.2 m/z +ve ionization, consistent with the expected mass of 433.93 g/mol.

I. FT-IR of Compound (1) HC1 salt Form A

The FT-IR spectrum of Compound (1) HC1 salt Form A is shown in FIG 21.

The FT-IR peaks for functional groups present for Compound (1) HC1 Form A are as shown in Table 6.

Table 6: FT-IR Peaks for Compound (1) HC1 Form A

Functional Group Wavenumber [cm⁴] Type

N-H 3483 medium, stretch

O-H 3202 broad, stretch

C-H (sp³) 2977 medium, stretch

C=O 1657 strong, stretch

C-Q 764 strong, stretch

EXAMPLE 6

Solubility studies of amorphous Compound (1) HC1 salt

The solubility of amorphous Compound (1) HC1 salt was assessed in 35 solvent systems.

Compound (1) HC1 salt (~700mg) was transferred to a 250mL round-bottom flask. To this, 50 mL of 1,4-dioxane: water (50:50 % v/v) was added and the solution stirred gently to promote dissolution. The solution was evenly split between 35 x 2mL vials (~1.4 mL solution; 20mg solid per vial). The vials were frozen and placed into a desiccator connected to a freeze drier and lyophilized for ca. 72 hours. A subsample was removed and analyzed by XRPD showing the material to be amorphous. To ca. 20mg of lyophilized amorphous material, the test solvent was added in 50pL aliquots. If solid remained the vial was gently heated to ca. 40 °C to aid dissolution. This cycle was repeated until the material fully dissolved or 2mL of solvent system had been added.

The approximate solubilities in 35 solvent systems investigated are shown in Table 7, along with the solvent’s International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) class. Amorphous Compound (1) HC1 was soluble in 16 solvents, partially soluble in 4 solvents, and insoluble in the remaining 15 solvents. High solubility was observed in methanol/water mixtures or pure methanol, acetic acid and dimethyl sulfoxide. Moderate solubility was observed in methanol/THF mixtures, 2- ethoxy ethanol, dichloromethane, ethanol, N,N- dimethylacetamide, N,N-dimethylformamide, nitromethane, N- methylpyrrolidone and low solubility was observed in chloroform.

After the assessment was complete, the vials (with both dissolved and undissolved material) were uncapped and evaporated at ambient temperature. Any solids produced through evaporation were analyzed by XRPD to determine form. Table 7 shows five forms were identified by XRPD analysis:

Form A identified from toluene;

Form B identified from 1,2-dimethoxyethane, 1 -butanol, 2-m ethyl THF, 2- propanol, acetone, acetonitrile, dichloromethane, ethyl acetate and methanol;

Form B with slight peak changes (possibly due to preferred orientation, denoted as “Form B + PO”) from ketones, propyl acetate, and methanol/THF mixtures;

Form D identified from 1,4-di oxane and THF;

Form E (consistent with Compound (1) Free Base) identified from water; and Form F identified from chloroform, a possible chloroform solvate.

The remaining samples did not produce sufficient solids for XRPD analysis.

Table 7: Solubility studies of amorphous Compound (1) HC1 salt e_e - Approximate Solubility XRPD Post- ICH

Sample Solvent / (mg/ /m _TL \) _c So ilu ib,i Tli*ty C--lIass

1 1,2-Dimethoxyethane 0 Form B 2

2 1,4-Dioxane 0 Form D 2

3 1 -Butanol 0 Form B 3 _{e e} - Approximate Solubility XRPD Post- ICH

Sample Solvent / (mg/ /m _TL \) _c So ilu ib,i Tli*ty C--lIass

4 2-Ethoxyethanol 25 no solid 2

5 2 -Methyl THF 0 Form B U

6 2-Propanol 0 Form B 3

7 Methanol: Water (40:60 % v/v) 5 no solid N/A

8 Methanol: Water (80:20 % v/v) 100 Form A N/A

9 Methanol: Water (95:5 % v/v) 200 mix A + B N/A

10 Acetic Acid 200 no solid 3

11 Acetone 0 Form B 3

12 Acetonitrile 0 Form B 2

13 Anisole 5 mix A + B 3

14 Chloroform 7 Form F 2

15 Dichloromethane 11 Form B 2

16 Dimethylsulfoxide 100 no solid 3

17 Ethanol 10 no solid 3

18 Ethyl Acetate 0 Form B 3

19 Heptane 0 mix A + B 3

20 Isopropyl Acetate 5 no solid 3

21 Methanol 100 Form B 2

22 Methylethyl Ketone 0 Form B + PO 3

23 Methylisopropyl Ketone 0 Form B + PO U

24 N,N'-Dimethylacetamide 14 no solid 2

25 N,N'-Dimethylformamide 50 no solid 2

26 Nitromethane 22 no solid 2

27 N-Methylpyrrolidone 50 no solid 2

28 Propyl Acetate 0 Form B + PO 3

29 tert-Butylmethyl Ether 0 mix A + B + D 3

30 THF 0 Form D 2

31 Toluene 5 Form A 2

32 Water 0 Form E N/A

33 Methanol: THF (75:25 % v/v) 67 Form B + PO N/A

34 Methanol: THF (50:50 % v/v) 67 Form B + PO N/A

35 Methanol: THF (25:75 % v/v) 50 Form B + PO N/A EXAMPLE 7

Maturation studies of amorphous Compound (1) HC1 salt

To amorphous Compound (1) HC1 salt, 100 pL aliquots of test solvent / solvent system were added until a mobile slurry was produced, and initial observations recorded. The vials were then capped, sealed with parafilm and temperature cycled for ca. 72 hours in 4-hour cycles between ambient (ca. 22 °C) and 40 °C with constant agitation. At this time all samples presented as off-white solids, other than THF: 0.01 N HC1 (50:50 % v/v), which produced an orange solid and solution. The solids were separated by centrifugation and analyzed by XRPD. The samples were then dried for ca. 24 hours at 40 °C and the XRPD analysis repeated. The results are presented in Table 8.

The saturated mother liquors were separated into three equal parts and retained for subsequent experiments, i.e. evaporation, crash cooling and anti-solvent addition (Examples 9, 10, and 11).

Table 8: Maturation Experiments

Solvent Solvent Volume Initial XRPD Post- XRPD Post-

(pL) Observations Maturation Drying

1,2-Dimethoxyethane 3500 milky slurry + solid contains B contains B

1,4-Dioxane 3000 milky slurry + solid Form D B + D

2-Ethoxyethanol 250 slurry Form B Form B

2-Methyl THF 3000 milky slurry + solid Form G Form G

2-Propanol 1000 milky slurry + solid Form B Form B

Methanol : Water ... . . .

(40 60 v/v) 1000 milky slurry amorphous amorphous

Methanol : Water , ,,,, . _ . _ .

/on o/ / \ 199 slurry Form A Form A

(80:20 % v/v)

Acetone 2000 milky slurry + solid Form B Form B

Solvent Solvent Volume Initial XRPD Post- XRPD Post-

(pL) Observations Maturation Drying

THF : 0.01 N HCl ... . . _c _

. 300 milky slurry A + E A + E

(25:75 % v/v)

EXAMPLE 8

Evaporation studies of amorphous Compound (1) HC1 salt

Evaporation experiments using the saturated solutions prepared from the maturation cycles described in Example 7, were set-up as follows: The saturated solutions were transferred to 2 mL vials, which were left uncapped and allowed to evaporate under ambient conditions. Observations were taken and any resulting solids were isolated by centrifugation and analyzed by XRPD, see Table 9, where X means no solids were isolated, (most samples did not produce sufficient solids for analysis.)

Table 9: Evaporation Experiments - observations and XRPD results e . . Observations Post- x

Solvent .. . XRPD

Evaporation

1,2-Dimethoxyethane X X

1,4-Dioxane off-white solid Form B

2-Ethoxyethanol off-white solid Form B

2 -Methyl THF X X

2-Propanol off-white solid Some B peaks

Methanol : Water (40:60 % v/v) off-white solid Form B

Methanol : Water (80:20 % v/v) off-white solid B & D

Acetone X X

Anisole X X

Chloroform X X

Dimethylsulfoxide off-white solid amorphous

Ethanol off-white solid A & B

Ethyl Acetate off-white solid amorphous

Isopropyl Acetate X X

Methanol X X Methylethyl Ketone X X

Methylisopropyl Ketone X X

N-Methylpyrrolidone X X

Propyl Acetate X X tert-Butylmethyl Ether X X

THF X X

Toluene X X

Water X X

Methanol : THF (50 : 50 % v/v) X X

THF : 0.01 N HC1 (50:50 % v/v) off-white solid Form A

THF : 0.01 N HC1 (25:75 % v/v) off-white solid Form A

EXAMPLE 9

Crash Cooling studies of amorphous Compound (1) HC1 salt

Crash cooling experiments using the saturated solutions prepared from the maturation cycles described in Example 7, were set-up as follows: The saturated solutions were transferred to 2 mL vials, which were capped and placed at 2-8 °C for ca. 72 hours.

Observations were then taken, and any resulting solids isolated by centrifugation and analyzed by XRPD. The samples were then placed in a freezer at < -10 °C for ca. 24 hours. Observations were then taken, and any resulting solids isolated by centrifugation and analyzed by XRPD as shown in Table 10, where X means no solids were isolated.

Table 10: Crash Cooling Experiments

„ , Observations Post- Observations Post-

^Solven' Cooling (5 °C) ^XRPD Cooling (-25 °C) ^XRPD

1,2-Dimethoxyethane clear solution X clear solution X

1,4-Dioxane frozen X X X

2-Ethoxyethanol off-white solid some B peaks X X

2-Methyl THF clear solution X clear solution X

2-Propanol very little solid X very little solid X

Methanol: Water (40:60 % v/v) clear solution X very little solid X _e , Observations Post- _vrinr. Observations Post-

^S°^lven' Cooling (5 °C) ^XRPD Cooling (-25 °C) ^XRPD

Methanol: Water (80:20 % v/v) very little solid X very little solid X

Acetone clear solution X very little solid X

Anisole clear solution X very little solid X

Chloroform clear solution X clear solution X

Dimethylsulfoxide pale orange solution X pale orange oil X

Ethanol off-white solid weak B X X

Ethyl Acetate clear solution X clear solution X

Isopropyl Acetate clear solution X clear solution X

Methanol clear solution X clear solution X

Methylethyl Ketone clear solution X very little solid X

Methylisopropyl Ketone clear solution X very little solid X

N-Methylpyrrolidone off-white slurry X X X

Propyl Acetate clear solution X clear solution X tert-Butylmethyl Ether clear solution X very little solid X

THF orange solution X orange oil X

Toluene clear solution X clear solution X

Water clear solution X frozen X

Methanol: THF (50:50 % v/v) orange solution X yellow oil X

THF: 0.01 N HC1 (50:50 % v/v) off-white solid Form A X X

THF: 0.01 N HC1 (25:75 % v/v) clear solution X frozen amorphous

EXAMPLE 10

Anti-solvent addition studies of amorphous Compound (1) HC1 salt

Anti-solvent experiments using the saturated solutions prepared from the maturation cycles described in Example 7, were set-up as follows: The saturated solutions were transferred to 2 mL vials and the appropriate antisolvent was added until either a change was observed or 1 mL had been added. Observations were taken of these changes. The samples were then capped and allowed to mature under ambient conditions for ca. 24 hours. Further observations were taken, and any resulting solids isolated by centrifugation and analyzed by XRPD as shown in Table 11, where X means no solids were isolated, and sm means small amount. Table 11: Anti -solvent Experiments - observations and XRPD results

EXAMPLE 11

Characterization of Crystal Forms B. D. E. F, G. and H

The various forms were characterized by TG/DTA, ¹ H NMR and PLM where possible. Origin of materials used for analyses, including solvent and experimental source, is provided in Table 12. Table 12: Primary Screen Material - Sources and Solvents

A. Form B

The material appeared crystalline by XRPD.

TG analysis showed no solvent related mass loss, due to the anhydrous nature of Form B.

DT analysis showed a melting endotherm with an onset at 189 °C and peak at

193 °C.

Large, rod-shaped crystals (ca. 20-200 pm) with birefringence were observed by

PLM. ¹H-NMR spectrum showed few impurities, with expected peaks (including the broad water peak at 3.1 ppm). No methanol was detected in the spectra however, a peak at ca. 4.11 ppm seen in Form A was absent.

B. Form D

The material appeared crystalline by XRPD.

TG analysis showed no solvent related mass loss. This is due to the anhydrous nature of Form D.

DT analysis showed a melting endotherm with an onset at ca. 186 °C and peak at ca. 191 °C. The melting point of Form D was slightly lower than previously observed with Form A.

Very fine crystals (ca. 1 - 5 pm) with irregular morphology and birefringence were observed by PLM.

¹H-NMR spectrum showed few impurities, with the expected peaks for Form A (including the broad water peak at 3.1 ppm). No THF peaks were observed in the spectra.

C. Form E

XRPD diffractogram of Form E was consistent with the diffractogram of Freebase material, indicating that Compound (1) HC1 disproportionates into the free base in water.

TG analysis showed no solvent related mass loss.

DT analysis showed a sharp melt endothermic event with an onset at ca. 185 °C and peak at ca. 187 °C. The melting point of Form E was slightly lower than Form A.

D. Form F

The material was crystalline by XRPD.

TG analysis showed a 17.58% solvent related mass loss from the onset of heating to ca. 150 °C, equivalent to 0.8 equiv. of chloroform, suggesting Form F is a chloroform solvate.

DT analysis showed a melt endothermic event with an onset at ca. 191 °C and peak at ca. 195 °C, followed by thermal decomposition.

¹H-NMR spectrum showed connectivity consistent with Form A (including the broad water peak at 3.1 ppm). Chloroform was detected by a singlet at 8.32 ppm, which integrated to 0.45, equivalent to 10.25% by weight. The discrepancy between TG/DTA and 1H NMR solvent levels is possibly due to dryness of the respective material batches used for these analyses.

Form F was found to be a metastable form, which converted to Form B upon drying under ambient conditions (both under vacuum and on air) or at 40 °C. Form F was confirmed to be a chloroform solvate.

E. Form G

The material was crystalline by XRPD.

TG analysis showed no significant mass loss in the solvent related mass loss area.

DT analysis showed a melting endotherm with an onset at ca. 185 °C and peak at ca. 190 °C, followed by thermal degradation. The melt is slightly lower than Form A.

Fine, rod-shaped crystals (ca. 10 - 20 pm) exhibiting some agglomeration and birefringence were observed by PLM.

¹H-NMR showed few impurities, with peaks consistent Form A (including the broad water peak at 3.1 ppm). Traces of solvent were detected: MEK singlet at ca. 2.07 ppm, integrating to 0.004, equivalent to 0.02% by weight.

F. Form H

The material was determined to be crystalline by XRPD.

TG analysis showed a 10.01% solvent related mass loss from the onset of heating to ca. 130 °C, equivalent to 0.48 equiv. of anisole. This suggested Form H may be an anisole solvate.

DT analysis showed a shallow, broad endothermic event with an onset of ca.

64 °C related to solvent loss, as well as a melt endothermic event with an onset at ca. 192 °C and peak at ca. 195 °C, followed by thermal decomposition. Form H may have desolvated at ca.

130 °C.

DSC analysis performed on Form H material previously heated to 150 °C with TG/DT instrument, showed a mass loss from the onset of heating up to ca. 130 °C related to solvent loss (loss of 0.5 equiv. of anisole). DT analysis showed a shallow, broad endothermic event with an onset at ca. 52 °C related to solvent loss, after which the curve returned to the baseline indicating solvent loss was complete.

DSC analysis of the post-TG/DT material showed no major endothermic events below 130 °C. A sharp, intense melting endotherm was observed from an onset at ca. 193 °C and peak at ca. 196 °C. The absence of solvent loss related endothermic events indicated this to be the melting point of the desolvated material.

VT-XRPD analysis of Form H suggested that upon heating Form H converts to a mixture of forms D and G. Where initially Form H was found to convert to predominantly Form G, an increased energy supply lead to a multi-step conversion into predominantly Form D.

¹H-NMR showed few impurities, with the expected peaks, including the broad water peak at 3.1 ppm). An anisole singlet was observed at 3.75 ppm, integrating to 0.49 (equivalent to 3.62% by weight).

EXAMPLE 12

Further Analyses of Forms B. D. and G

Due to their desirable physical properties, Forms B, D and G were identified for scale up. Solvent choice was based on previous results, including crystallinity of produced material and stability. Each material was prepared on a 1g scale and characterized by XRPD, PLM, TG/DTA, DSC, DVS, FT-IR, 1H NMR, UPLC, 7-day stability, and VT-XRPD (Form B).

Scale-Up Procedure: To each of four or five scintillation vials containing ca. 240mg of amorphous material solvent was added to create a mobile slurry. The slurry was temperature cycled with agitation between ambient (ca. 22 °C) and 40 °C for ca. 72 hours to provide a milky slurry containing solid material. A subsample of the matured slurry was analyzed by XRPD. The solid material was isolated using Buchner filtration apparatus, collected, weighted and dried under vacuum at 40 °C for ca. 72 hours. The dried material was analyzed by XRPD (in each case the same form was obtained before and after isolation & drying). The vials were reweighted and gravimetric % yield calculated. A summary of the scale-up results is presented in Table 13. A gravimetric yield was calculated based on the weight of solids recovered, with % yields presented in Table 14. Table 13: Maturation Experiments Results for Forms B, D and G

IPA: 2-propanol; MEK: Methylethyl ketone

* Recorded twice - before and after isolation & drying; in each case the same Form was observed

Table 14: Yields (%) for Forms B, D and G.

A. Form B further characterization

Compound (1) HC1 produced crystalline material with an XRPD diffraction pattern consistent with previously identified Form B (FIG 22), observed diffraction peak listings are provided in Table 15. Table 15: XRPD Peak List of Compound (1) HC1 salt Form B

PLM analysis showed the material to have a form of fine (ca. 5-20 pm), birefringent crystals with irregular morphology and agglomeration. TG analysis (FIG 23) showed no solvent related mass loss, which correlates with previous results and indicates Form B is an anhydrous form. DT analysis showed a sharp melting endotherm with an onset at ca. 193 °C and peak at ca. 195 °C, followed by thermal decomposition. DSC analysis (FIG 24) showed no significant endothermic events related to solvent loss. An intense, sharp melting endotherm was observed with an onset at ca. 196 °C and peak at ca. 198 °C. Form B demonstrated the highest melting point compared to the other forms observed. No significant thermal events were noted in the cooling or second heating cycles. DVS analysis (FIG 25)showed Form B to be slightly hygroscopic, with a maximum mass increase of 0.12% wt. at 90% RH (during first sorption cycle). Form B demonstrated the lowest hygroscopicity compared to other forms observed. The material dehydrated slowly during the desorption cycle until ca. 20% RH. Post-DVS XRPD 20 diffractogram showed there were no changes to the material lattice after exposure to the DVS humidity conditions. DVS analysis at 40 °C showed Form B to be slightly hygroscopic, with maximum mass increase at 90% RH (during first sorption) to be 0.18% wt. Maximum mass increase at 40 °C (at 90%RH during first sorption) was 0.06% higher than during DVS analysis at ambient temperature. After the initial bigger mass gain during first sorption, material lost more mass during desorption stages and gained comparatively less mass during the second sorption. Post-DVS (40 °C) XRPD 20 diffractogram showed no changes to the Form B structure occurred during exposure to the DVS humidity conditions.

¹H-NMR spectrum of Form B (FIG 26) has peaks consistent with spectrum of

Form A, with only minimal impurities detected. Traces of 2- Propanol were observed at ca. 1.04 ppm, which integrated to 0.13% weight. An FT-IR spectrum (FIG 27) of Form B was taken and major functional group peaks identified in Table 18. HPLC showed the material is 99.9% pure.

B. Form D further characterization Compound (1) HC1 produced crystalline material with an XRPD diffraction pattern (FIG 28) consistent with previously identified Form D, observed diffraction peak listings are provided in Table 16.

Table 16: Diffractogram Peak List of Form D

PLM analysis showed agglomerates of fine (ca. 1-20 gm), birefringent crystals with irregular morphology. TG analysis (FIG 29) showed no solvent related mass loss, which correlates with previous results and suggests Form D was an anhydrous form. DT analysis showed a sharp melt endothermic event with an onset at ca. 191 °C and peak at ca. 194 °C, followed by thermal decomposition. DSC analysis (FIG 30) showed no significant endothermic events related to solvent loss. An intense, sharp melt endotherm was observed with an onset at ca. 192 °C and peak at ca. 196 °C. Form D appeared to have the second highest melting point compared to other forms observed. No significant thermal events were observed during the cooling or second heating cycles. DVS analysis (FIG 31) showed Form D to be slightly hygroscopic (class II hygroscopicity), with maximum mass increase at 90%RH (during first sorption) to be 0.8%. Post-DVS XRPD 20 diffractogram showed no significant changes in form after exposure to the DVS humidity conditions. NMR spectrum (FIG 32) of Form D is consistent with spectrum of Form A, with only minimal impurities detected. No solvent peaks were detected. An FT-IR spectrum of Form D (FIG 33) was taken and major functional group peaks identified in Table 18. HPLC showed the material is 99.7% pure.

C. Form G further characterization

Compound (1) HC1 produced crystalline material with an XRPD diffraction pattern (FIG 34) consistent with previously identified Form G, observed diffraction peak listings are provided in Table 17. Table 17: Diffractogram Peak List of Form G

PLM analysis showed the material to have medium (ca. 10-50 gm), needle- shaped, birefringent crystals and agglomerates of crystals. TG analysis (FIG 35) showed no solvent related mass loss, which corelates with previous results and suggests Form G to be an anhydrous form. DT analysis (FIG 36) showed a comparatively weaker melting endotherm with an onset at ca. 188 °C and peak at ca. 190 °C, followed by thermal decomposition, with no significant endothermic events related to solvent loss. An intense melt endothermic event was observed with an onset at ca. 189 °C and peak at ca. 195 °C. Form G appeared to have the lowest melting point of the prepared forms. No significant thermal events were noted in the cooling or second heating cycles. DVS analysis (FIG 37) showed Form G to be slightly hygroscopic (class II hygroscopicity), with maximum mass increase at 90% RH (during first sorption) to be 0.23% wt. Form G was slightly more hygroscopic than Form B. The material exhibited a slower mass loss during desorption, which slowed down further under 40% RH. Post-DVS XRPD 20 diffractogram showed there were no significant changes in form after exposure to the DVS humidity conditions. ¹H NMR spectrum (FIG 38) of Form G is consistent with spectrum of Form A, with only minimal impurities detected. Traces of solvent were detected: MEK triplet at ca. 0.91 ppm, integrating to 0.07 (equivalent to 0.36% by weight). An FT-IR spectrum (FIG 39) of Form G was taken and major functional group peaks identified in Table 18. HPLC showed the material is 99.7% pure.

Table 18: FT-IR peaks for functional groups present for Forms B, D and G.

Polarized Light Microscopy (PLM) Images of Forms A, B, D and G are shown in

FIG 40.

EXAMPLE 13

1-Week Stability Study (Forms B, D & G) Approximately lOmg of material was weighted into 18 x 2mL vials (6 vials per

Form). Two vials with each form were stored under the following conditions for 1 week, after which time, the solids were analyzed by XRPD and HPLC to determine purity; the results are presented in Table 19.

Ambient temperature, light, and humidity (capped vials) 40 °C / 75 %RH (uncapped vials)

80 °C (capped vials)

Table 19: One Week Stability Study Results

Form B was stable across all conditions investigated.

Form D was stable at ambient conditions though converted to Form B at increased temperature and/or increased relative humidity.

Form G was stable at ambient conditions and as predominantly Form G at increased temperature / relative humidity with some Form D and B peaks observed.

EXAMPLE 14

Solubility in HC1 (0. IN and 0.01N) (Forms B, D & G)

The solubility of Forms B, D and G was assessed in 0.1N HC1 and 0.01N HC1. Approximately 30mg of Forms B and D and ca. 40mg of Form G was weighed into x 12 scintillation vials (2 sets of 6 vials). ImL of appropriate buffer was added to create a mobile slurry. The first set of 6 vials were agitated for 4 hours and the second for 24 hours. After this time, the solids were isolated by centrifugation with the use of a nylon filter and analyzed by XRPD. The mother liquors were analyzed by HPLC to determine concentration. The results are presented in Table 20, and show all solids converted into Form A, (other than Form B after 4 hours in 0. IN HC1, which retained its original form.) Solubility of ca. 5 mg/mL was identified in the samples agitated for 4 hours in 0.1 N and 0.01 N HC1 Form B (5.1 and 5.5 mg/mL respectively). All others presented solubility between 2.1 and 2.7 mg/mL.

Table 20: Observations and Results from the Solubility Tests in HC1

EXAMPLE 15

Stability Assessment: Competitive Slurrying (Forms B, D & G)

An assessment of the thermodynamic stability between Forms B, D and G was performed. Approximately lOmg of each form (B, D and G per each vial; 30mg total) was weighted into four vials. lOOpL of solvent system, either 50:50 or 25:75 methanol:THF (% v/v), was added to each vial to provide an off-white mobile slurry. A stirrer bar was added, and the vials stirred at either ambient (ca. 22 °C) or 60 °C for ca. 48 hours. The solids were collected by centrifugation with the use of a nylon filter and analyzed by XRPD and the results shown in Table 21. Form B was determined to be the most stable polymorph.

Table 21: Stability Assessment: Competitive Slurrying

EXAMPLE 16

Water Activity of Form B in MeOH (aw 0.3 and 0.5)

The stability of Form B was examined at two water activity levels (a_w 0.3 and 0.5) in methanol. Form B material was weighed into 4 vials (ca. 30 mg / vial). 150pL of appropriate solvent system (in 3 x 50pL aliquots) was added to each vial to provide off-white mobile slurries. The vials were stirred at either ambient (ca. 22 °C) or 60 °C for ca. 48 hours, then solids collected by centrifugation and analyzed by XRPD. The results are shown in Table 22.

0.3 a_w methanol: add 0.1 mL deionized water per 1 mL of solution 0.5 a_w methanol: add 0.23 mL deionized water per 1 mL of solution.

Table 22: Water Activity (a_w) of Form B

Compound (1) Salts

Several attempts were made to prepare the Compound (1) xinafoate, 3-hydroxy-2- naphthoate, napsylate and calcium salts. Initial attempts were unsuccessful, with Compound (1) Free Base Form 1 isolated.

Further attempts, under varying conditions provided a novel napsylate salt; a xinafoate and 3-hydroxy-2-naphthoate salt. Xinafoate Forms 1 and 2 were isolated from di chloromethane and ethyl acetate respectively. 5g scale up preparation of napsylate and xinafoate Form 1 salts were successful. The solids isolated were highly crystalline, with high purity, and demonstrated 1 equivalent of counterion by NMR. All attempts to form the calcium salt were unsuccessful. EXAMPLE 17

Initial Compound (1) Salt Preparations

A. Compound (1) Xinafoate salt (Methanol)

Compound (1) Free Base (2 g) and methanol (124 mL) were gently heated to approximately 60 °C, with sonication, to form a slurry. A slurry of l-hydroxy-2-naphthoic acid (4.6 mL, ca 1 eq) was added and the mixture heated at 40 °C with agitation for approximately 10 minutes. A sample was removed by centrifugation filtration, which was Compound (1) Free Base Form 1 by XRPD.

The slurry was heated at 50 °C with stirring (300 rpm) for 1 hour, during which time the slurry become a clear solution. Upon cooling to ambient temperature (ca 40 minutes) a precipitate formed. A sub sample was taken and analysed by XRPD and confirmed to be Compound (1) Free Base Form 1. The mixture was stirred for a further 16 hours and heated to 50 °C resulting in redissolution of solids to form a clear solution. Additional 1 -hydroxy-2 - naphthoic acid solution (2.3 mL, ca 0.5 eq) was added and the mixture cooled to ambient temperature. Methanol was removed under vacuum to provide a light brown solid which was analysed by XRPD. A new, very weakly crystalline diffractogram was obtained.

B. Compound (1) Xinafoate salt (THF)

A mixture of Compound (1) Free Base (2 g) in THF (40 mL) was heated at 40 °C with agitation until dissolution was complete, l-hydroxy-2-napthoic acid (894.15 mg, 1 eq) was added and the temperature cycled between ambient and 40 °C in 4-hour cycles for 72 hours to provide a clear, darkened solution. The volume of THF was reduced to ca. 10 mL and after 10 minutes standing at room temperature (22 °C) a precipitate was observed, which was determined by XRPD to be Compound (1) Free Base Form 1. Fresh THF (10 mL) was then added with agitation and heating to approximately 40 °C until a clear solution formed. The solution was then placed at 2-8 °C for 24 hours resulting in the formation of a slurry, which was determined by XRPD to be Compound (1) Free Base Form 1. The bulk solids were filtered and dried under vacuum at ambient temperature; the mother liquor was concentrated under vacuum to provide a viscous oil. The isolated solids were determined by XRPD to be Compound (1) Free Base Form 1. ¹H NMR of the final solids showed no evidence of salt formation.

C. Compound (1) 3 -Hydroxy-2 -Naphthoate salt

A mixture of Compound (1) Free Base (2 g) in THF (40 mL) was heated at 40 °C with agitation until dissolution was complete. 3-hydroxy-2-napthoic acid (894.22mg, 1 eq) was added and the temperature cycled between ambient and 40 °C in 4-hour cycles for 72 hours. The volume of THF was reduced to ca. 10 mL and heptane (2 mL) added as an anti-solvent dropwise until a slurry formed. The slurry solids were confirmed to be Compound (1) Free Base Form 1 by XRPD. The slurry was then placed at 2-8 °C for ca. 24 hours, fresh THF (10 mL ) added and then heated at 40 °C with agitation until a clear solution formed. The sample was then again placed at 2-8 °C overnight to yield a slurry, which was determined by XRPD to be Compound (1) Free Base Form 1.

The bulk solids were filtered and dried under vacuum at ambient temperature; the mother liquor was concentrated under vacuum to provide a viscous oil. ¹H NMR analysis of the final solids isolated showed no evidence of salt formation.

D. Compound (1) Napsylate salt

A mixture of Compound (1) Free Base (2 g) in THF (40 mL) was heated at 40 °C with agitation until dissolution was complete. Naphthalene-2-sulfonic acid (989.05 mg, 1 eq) was added and the temperature cycled between 20 °C and 40 °C in 4-hour cycles for 72 hours. The volume of THF was reduced to ca. 10 mL and heptane (2 mL) added dropwise. No solids were formed. The solution was placed at 2-8 °C overnight resulting in the formation of a slurry. The slurry solids were confirmed to be Compound (1) Free Base Form 1 by XRPD. The bulk solids were filtered and dried under vacuum at ambient temperature; the mother liquor was concentrated under vacuum to provide a viscous oil. ¹H NMR analysis of the final solids isolated showed no evidence of salt formation.

E. Compound (1) Calcium Salt

A mixture of Compound (1) Free Base (2 g) in THF (40 mL) was heated at 40 °C with agitation until dissolution was complete. Calcium hydroxide (352.23 mg, 1 eq) was added and the temperature cycled between ambient and 40 °C in 4-hour cycles for 72 hours resulting in the formation of a slurry. The slurry solids were confirmed to be Compound (1) Free Base Form 1 by XRPD. The slurry was stored at ambient for 12 hours and then filtered. The dried isolated solids were weakly crystalline Compound (1) Free Base Form 1 by XRPD. THF (3 mL) was added to the isolated solids to form a mobile slurry which was shaken at ambient conditions for 1 hour and the solids again isolated by filtration, which were amorphous. Methanol (3 mL) was added to the isolated solids and the resulting slurry heated to 40 °C. The solids were isolated by filtration and were amorphous.

Table 23: Summary of initial salt preparation attempts (ML = mother liquor) - 68 5

EXAMPLE 18

Further Compound (1) Salt Preparations

A. Compound (1) Xinafoate salt

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at

40 °C with agitation until dissolution was complete, l-hydroxy-2-napthoic acid (227.53 mg, 1.05 eq) was added resulting in a darkening of the clear solution. The solution was placed at 2-8 °C for ca. 24 hours, resulting in the formation of a precipitate. The solids were isolated by filtration and shown to be Compound (1) Free Base Form 1 by XRPD. B. Compound (1) 3 -Hydroxy-2 -Naphthoate salt

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at 40 °C with agitation until dissolution was complete. 3-hydroxy-2-napthoic acid (227.60 mg, 1.05 eq) was added resulting in the clear solution turning slightly yellow. The solution was placed at 2-8 °C for ca. 24 hours, resulting in the formation of a precipitate. The solids were isolated by filtration and shown to be Compound (1) Free Base Form 1 by XRPD.

C. Compound (1) Napsylate salt

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at 40 °C with agitation until dissolution was complete. Naphthalene-2-sulfonic acid (251.71 mg, 1.05 eq) was added resulting in the clear solution turning pale pink. The solution was placed at 2- 8 °C for ca. 24 hours during which the solution darkened though no solids had formed. Heptane (500 pL) was added dropwise at ambient temperature (22 °C) until precipitation was observed and the solids isolated by filtration.

XRPD analysis identified a new diffraction pattern different from both the free base pattern 1 and the diffraction pattern for 2-naphthalenesulfonic acid. PLM showed small, birefringent particles with no clearly defined morphology. ¹ H-NMR showed evidence of salt formation from shifting peaks and a broadened water peak with 1 equivalent of naphthalene-2- sulfonic acid present.

FT-IR analysis showed peak shifting indicating formation of a salt.

1691cm'¹ - C=O stretching vibration (carbonyl and ester functionality) 3057cm'¹ - R-OH stretching vibration.

D. Compound (1) Calcium salt (using excess of free base)

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at 40 °C with agitation until dissolution was complete. Calcium hydroxide (82.18 mg, 0.95 eq) was added to yield a thick off-white slurry which was filtered to produce a damp solid. The dried solid was shown be amorphous by XRPD. THF (5 mL) was added to the dried solid to form a slurry. Heptane (2.5 mL) was added and the temperature cycled between 20 and 40 °C in four- hour cycles for ca. 16 hours. The slurry was filtered, and the mother liquor retained. Heptane (4 mL) was added to the mother liquor and additional precipitation was observed. The isolated solids were dried and analyzed.

XRPD analysis showed the solids isolated from the mother liquor were amorphous. PLM analysis showed very small, non-birefringent particles with no clearly defined morphology. ¹H-NMR showed a significant increase in the number of peaks, peak broadening and peak shifting, signifying decomposition. CAD analysis found the material to contain 4.3 %w/w calcium, approximately equivalent to a hemi calcium salt. HPLC analysis found the material to be 88.8 % by % area.

E. Compound (1) Calcium salt (using 1 or 0.5 Equivalents Ca) A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at

40 °C with agitation until dissolution was complete. Calcium hydroxide (85.50 mg, 1 eq or 42.68 mg, 0.5 eq) was added to form an off-white slurry. Solids were isolated by filtration, dried and analyzed by XRPD which showed the material to be amorphous. ¹H-NMR, near identical spectra obtained for both experiments, showed signs of degradation with significant peak broadening and peak shifts.

Table 24: Summary of Further Salt Preparations

EXAMPLE 19

Additional Xinafoate and 3 -Hydroxy-2 -Naphtoate Crystallizations

Compound (1) Free Base (ca 50 mg) was weighed into 10 x 2 mL vials and solvent (500 pL) added to each vial. Either solid l-hydroxy-2-naphthoic acid (1.05 eq) or 3- hydroxy-2-naphthoic acid stock solution (1.05 eq) was added to the vials, which were then heated at 40 °C for ca. 1 hour and cooled to room temperature (approximately 20 °C). Solids were isolated by centrifuge filtration and analyzed by XRPD.

Material isolated from 3-hydroxy-2-naphthoate was Compound (1) Free Base Form 1 in all cases. XRPD results for Xinafoate are shown in Table 25, where Xinafoate salts were observed from DCM and ethyl acetate.

Table 25: Experimental Details for Compound (1) Xinafoate salt

EXAMPLE 20

Compound (1) Salt Formation alternate conditions A. Compound (1) Napsylate

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at

40 °C with agitation until dissolution was complete. Naphthalene-2-sulfonic acid (359.80 mg, 1.05) was added and the mixture heated at 40 °C for 1 hour, cooled to 20 °C and then held at 2- 8 °C for 16 hours to form a pale off white slurry. Solids were isolated by filtration, dried and characterized by XRPD, NMR, TGA/DSC, FT-IR, PLM and HPLC.

XRPD showed a diffraction pattern consistent with the previous preparation, designated Compound (1) Napsylate salt, Form 1, FIG 41.

¹H-NMR analysis (FIG 42) confirmed salt formation, with 1 equivalent of naphthalene-2-sulfonic acid, which was 99.6 % pure by HPLC. PLM showed the material consisted of fine, birefringent particles with some defined needle-like morphology.

TGA/DSC analysis (FIGS 43 and 44) revealed a gradual initial mass loss of 2.8 wt. %, most likely due to loss of 0.5 equivalents of water. Mass loss of 18.7 wt. % with onset at 120 °C was due to loss of l-hydroxy-2-naphthoic acid. Weak melt onset 110 °C with degradation evident above 310 °C.

FT-IR analysis (FIG 45) showed evidence of salt formation. The FT-IR peak list is shown in Table 26.

Table 26: Compound (1) Napsylate salt FT-IR peak list

B. Compound (1) Xinafoate Form 1

A mixture of Compound (1) Free Base (500mg) in DCM (5 mL) was heated at 40 °C with agitation until dissolution was complete, l-hydroxy-2-naphthoic acid (234.79 mg, 1.05) was added and the mixture heated at 40 °C for 1 hour, and then cooled to 20 °C to provide a pale clear solution. Heptane (4 mL) was added dropwise until precipitation was observed and the mixture then placed at 2-8 °C for ca. 16 hours, to provide a thick immobile slurry. Dichloromethane : heptane 1 : 1 (5 mL) was added and the mixture heated at 40 °C for one hour resulting in re-dissolution. Heptane (4 mL) was added to the clear solution until the solution appeared turbid, and then placed at 2-8 °C for ca. 16 hours, again providing a thick slurry. Solids were isolated by filtration, dried and characterized by XRPD, NMR, TGA/DSC, FT-IR, PLM and HPLC.

XRPD (FIG 46) showed a diffraction pattern consistent with the previous preparation, designated Compound (1) Xinafoate salt, Form 1. PLM showed the material consisted of fine, birefringent particles with some defined needle-like morphology.

¹H-NMR analysis (FIG 47) confirmed salt formation, with 1 equivalent of 1- hydroxy-2-naphthoic acid, which was 98.8% pure by HPLC.

TGA/DSC (FIGS 48 and 49) analysis revealed a large mass loss of 49 wt.% with onset 120 °C in the TGA, likely due to loss of counterion and degradation. There was a melting endothermic event with onset at 100 °C in the DSC. Additional endothermic events, corresponding to loss of counterion (onsets at 130 °C and 167 °C) were also present. Degradation was evident above 270 °C.

FT-IR (FIG 50) analysis showed evidence of salt formation. The FT-IR peak list is shown in Table 27. Table 27: Compound (1) Xinafoate salt Form 1 FT-IR peak list

C. Compound (1) Xinafoate Form 2

Compound (1) Free Base (500 mg) and ethyl acetate (5 mL) were heated at 40 °C with agitation, to provide a slurry, l-hydroxy-2-naphthoic acid (234.44 mg, 1.05 eq) was added and the mixture heated at 40 °C for 1 hour. After cooling to 20 °C a thick immobile slurry formed to which ethyl acetate (5 mL) was added. The mixture was again heated at 40 °C for a further hour, and cooled to 20 °C. The solids from the resulting slurry were isolated, dried and characterized by XRPD, NMR, TGA/DSC, FT-IR, PLM and HPLC.

XRPD (FIG 51) showed a diffraction pattern consistent with the previous preparation designated xinafoate Form 2. PLM showed the material consisted of very small, fine, birefringent particles with some defined needle-like morphology.

¹H-NMR (FIG 52) analysis confirmed salt formation with 1 equivalent of 1- hydroxy-2-naphthoic acid which was 99.0 % by HPLC.

TGA/DSC (FIGS 53 and 54) analysis revealed a large mass loss of 49 wt.% onset 120 °C likely due to loss of counterion and degradation. Broad melt onset -100 °C and an endothermic event corresponding to loss of counterion onset 165 °C. Degradation evident above 260 °C.

FT-IR (FIG 55) analysis showed evidence of salt formation. The FT-IR peak list is shown in Table 28. Table 28: Compound (1) Xinafoate salt Form 2 FT-IR peak list

D. Compound (1) Calcium salt (from calcium hydroxide)

A mixture of Compound (1) Free Base (500mg) in THF (5 mL) was heated at

40 °C with agitation until dissolution was complete. Calcium hydroxide (82.74 mg, 1.05 eq) was added and the mixture heated at 40 °C for 1 hour, then cooled to 20 °C to provide a thick white slurry. The solids were isolated by filtration and found to be Compound (1) Free Base by XRPD. Heptane (2 mL) was added dropwise to the mother liquor and then placed at 2-8 °C overnight resulting in formation of a slurry. Solids were isolated by filtration and dried. Only

Compound (1) Free Base Form 1 was observed by XRPD and ¹H-NMR of the sample showed no evidence of salt formation. E. Compound (1) Calcium salt (from Calcium Methoxide)

Compound (1) Free Base (50 mg) was weighed into 7 x 2 mL vials. Solvent (ImL) was added and the mixtures heated to 40 °C until clear solutions formed. Calcium methoxide (1.05 eq) was added and the vials agitated at 20 °C for 14 days. The resulting material was analyzed by XRPD and ¹H-NMR. Table 29: Attempts to form Ca salt from CaOMe

¹H-NMR of amorphous solids showed high levels of degradation in the solids from ethyl acetate and methanol. Amorphous solids from di chloromethane showed no decomposition, but no clear evidence of salt formation.

EXAMPLE 21

Solubility of Compound (1) Salts in Simulated GI Fluids The solubility of the Compound (1) napsylate and xinafoate salts was determined in water and the following buffer systems:

FeSSIF Simulating Fed State Small Intestines

FaSSIF Simulating Fasted State Small Intestines

FaSSGF Simulating Fasted State Gastric (Stomach)

Compound (1) Free Base (ca. 5 mg) was weighed into a vial and buffer was added (50 pL aliquots up to 1 mL) to achieve a slurry. The vials were placed into a heating block and agitated at 37 °C (the slurries prepared in water were agitated at 25 °C). After 1 hour, 200 pL of each sample was filtered by centrifugation filtration, analyzed by HPLC and where possible XRPD. After 24 hours of agitation, a further 200 pL of each sample was filtered by centrifugation filtration, analyzed by HPLC and where possible XRPD. After 48 hours of agitation, 500 pL of each sample was filtered by centrifugation filtration analyzed by HPLC and where possible XRPD analysis, shown in Table 30. The solubility of the salts was low in all systems (see Table 31), decreased over time in SGF and fluctuated in the remaining buffer systems. Table 30: XRPD Results

* = insufficient solids

Am = Amorphous

XI = Compound (1) xinafoate Form 1 FBI = Weakly crystalline Compound (1) Free Base Form 1

Table 31: Compound (1) Free Base Concentration (pg/mL)

EXAMPLE 22

Compound (1) Xinafoate salt 5g scale preparation

Compound (1) Free Base (5.25 g) and DCM (50 mL) were heated at 40 °C to form a clear solution. l-Hydroxy-2-naphthoic acid (2467.40 mg, 1.05 eq) was added and the mixture heated at 40 °C for 1 hour and then allowed to cool to 20 °C to form a slightly darkened clear solution. Heptane (40 mL) was added until a precipitate formed, and the mixture then placed at 2-8 °C for ca. 16 hours to form a thick immobile slurry. DCM:heptane 1 : 1 mixture (20 mL) was added to mobilise the slurry and the solids were isolated by filtration, dried analyzed by XRPD, NMR and HPLC. The isolated material was Compound (1) xinafoate, Form 1 by XRPD. ^-NMR confirmed salt formation (1 equivalent of 1 -hydroxy-2 -naphthoic acid), with a purity of 98.82 % by HPLC.

EXAMPLE 23

Compound (1) Napsylate salt 5g scale preparation

Compound (1) Free Base (5.25 g) and DCM (50 mL) were heated at 40 °C to form a clear solution. Naphthalene-2-sulfonic acid (3779.27 mg, 1.05 eq) was added and the mixture heated at 40 °C for 1 hour during which time a precipitate formed. After cooling to ambient temperature, the mixture was placed at 2-8 °C overnight, resulting in the formation of a pale off white slurry. Solids were isolated by filtration, dried and analyzed by XRPD, NMR and HPLC. The isolated material was Compound (1) napsylate, Form 1 by XRPD. ¹H-NMR confirmed salt formation (1 equivalent of 2-naphthalene sulfonic acid), with a purity of 99.87% by HPLC.

The various embodiments described above can be combined to provide further embodiments. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and/or listed in the Application Data Sheet are incorporated herein by reference, in their entirety. Aspects of the embodiments can be modified, if necessary, to employ concepts of the various patents, applications and publications to provide yet further embodiments.

These and other changes can be made to the embodiments in light of the abovedetailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

Claims

1. A crystalline form of an HC1 salt of a compound having the following structure (1):

2. The crystalline form of claim 1, wherein the crystalline form of the HC1 salt of Compound (1) is anhydrous.

3. The crystalline form of claim 2 having an X-ray powder diffraction

(XRPD) spectrum comprising at least 3 peaks with diffraction angles (29 ± 0.5°) selected from

Table 15.

4. The crystalline form of claim 2 or 3 having an X-ray powder diffraction (XRPD) spectrum comprising at least 6 peaks with diffraction angles (29 ± 0.5°) selected from Table 15.

5. The crystalline form of any one of claims 2-4 having an X-ray powder diffraction (XRPD) spectrum comprising at least 10 peaks with diffraction angles (29 ± 0.5°) selected from Table 15.

6. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 16.8°, 18.6°, 15.2°, 9.0°, and 20.3°.

7. The crystalline form of claim 6, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 4.8°, 22.6°, 18.8°, 19.9°, and 20.7°.

8. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 16.8°, 18.6°, 15.2°, 9.9°, and 29.3°.

9. The crystalline form of claim 8, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 4.8°, 22.6°, 18.8°, 19.9°, and 29.7°.

19. The crystalline form of claim 2 having an XRPD spectrum substantially as depicted in Figure 22.

11. The crystalline form of any one of claims 3-1 , wherein the XRPD is measured using Cu K radiation in transmission mode using 49 kV / 49 mA generator settings.

12. The crystalline form of any one of claims 2-11 having a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting.

13. The crystalline form of any one of claims 2-12 having a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C.

14. The crystalline form of any one of claims 2-13 having a DTA thermogram having a melting isotherm with an onset at about 189 °C and a peak at about 193 °C.

15. The crystalline form of claim any one of claims 2-13 having a DTA thermogram having a melting isotherm with an onset at about 193 °C and a peak at about 195 °C.

78

16. The crystalline form of any one of claims 2-13 having a TG/DTA thermogram substantially as depicted in Figure 23.

17. The crystalline form of any one of claims 2-16 having a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting.

18. The crystalline form of any one of claims 2-17 having a DSC thermogram showing a melting endotherm with onset at about 196 °C and a peak at about 198 °C.

19. The crystalline form of any one of claims 2-18 having a DSC thermogram substantially as depicted in Figure 24A or Figure 24B.

20. The crystalline form of any one of claims 2-19 having a proton nuclear magnetic resonance ( ’H NMR) spectrum substantially as depicted in Figure 26.

21. The crystalline form of any one of claims 2-20 having a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 25.

22. The crystalline form of any one of claims 2-21 having a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2344 cm^-1, 2963 cm^-1, 3062 cm^-1, 3179cm^-1.

23. The crystalline form of any one of claims 2-22 having an FT-IR spectrogram substantially as depicted in Figure 27.

24. The crystalline form of claim 2 having an X-ray powder diffraction

(XRPD) spectrum comprising at least 3 peaks with diffraction angles (29 ± 0.5°) selected from Table 16.

25. The crystalline form of claim 2 or 24 having an X-ray powder diffraction

(XRPD) spectrum comprising at least 6 peaks with diffraction angles (29 ± 0.5°) selected from

Table 16.

26. The crystalline form of any one of claims 2, 24, or 25 having an X-ray powder diffraction (XRPD) spectrum comprising at least 10 peaks with diffraction angles (29 ± 0.5°) selected from Table 16.

27. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 4.3°, 9.0°, 16.9°, 18.1°, and 22.6°.

28. The crystalline form of claim 27, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 14.7°, 15.2°, 16.2°, 18.6°, and 19.3°.

29. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 4.3°, 9.0°, 16.9°, 18.1°, and 22.6°.

30. The crystalline form of claim 29, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 14.7°, 15.2°, 16.2°, 18.6°, and 19.3°.

31. The crystalline form of claim 2 having an XRPD spectrum substantially as depicted in Figure 28.

32. The crystalline form of any one of claims 27-31, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

80

33. The crystalline form of any one of claims 2 and 27-31 having a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting.

34. The crystalline form of any one of claims 2 and 27-31 having a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C.

35. The crystalline form of any one of claims 2 and 27-34 having a DTA thermogram having a melting isotherm with an onset at about 191 °C and a peak at about 194 °C.

36. The crystalline form of any one of claims 2 and 27-35 having a TG/DTA thermogram substantially as depicted in Figure 29.

37. The crystalline form of any one of claims 2 and 27-36 having a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting.

38. The crystalline form of any one of claims 2 and 27-36 having a DSC thermogram showing a melting endotherm with onset at about 192 °C and a peak at about 196 °C.

39. The crystalline form of any one of claims 2 and 27-36 having a DSC thermogram substantially as depicted in Figure 30A or Figure 30B.

40. The crystalline form of any one of claims 2 and 27-39 having a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 32.

41. The crystalline form of any one of claims 2 and 27-40 having a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 0.8% at 90% relative humidity during first sorption.

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42. The crystalline form of any one of claims 2 and 27-41 having a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 31.

43. The crystalline form of claim 41 or 42, wherein the crystalline form of the HC1 salt is class II hygroscopic.

44. The crystalline form of any one of claims 2 and 27-43 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2349 cm^-1, 2933 cm^-1, 3062cm^-1, and 3177 cm^-1.

45. The crystalline form of any one of claims 2 and 27-44 having an FT-IR spectrogram substantially as depicted in Figure 33.

46. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising at least 3 peaks with diffraction angles (29 ± 0.5°) selected from Table 17.

47. The crystalline form of claim 2 or 46 having an X-ray powder diffraction (XRPD) spectrum comprising at least 6 peaks with diffraction angles (29 ± 0.5°) selected from Table 17.

48. The crystalline form of any one of claims 2, 46, or 47 having an X-ray powder diffraction (XRPD) spectrum comprising at least 19 peaks with diffraction angles (29 ± 9.5°) selected from Table 17.

49. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 4.7°, 9.5°, 13.2°, 16.7°, and 19.3°.

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50. The crystalline form of claim 49, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 11.2°, 19.1°, 19.5°, 21.2°, and 24.5°.

51. The crystalline form of claim 2 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 4.7°, 9.5°, 13.2°, 16.7°, and 19.3°.

52. The crystalline form of claim 51, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 11.2°, 19.1°, 19.5°, 21.2°, and 24.5°.

53. The crystalline form of claim 2 having an XRPD spectrum substantially as depicted in Figure 34.

54. The crystalline form of any one of claims 49-53, wherein the XRPD is measured using Cu K radiation in transmission mode using 49 kV / 49 mA generator settings.

55. The crystalline form of any one of claims 2 and 49-54 having a thermogravimetric analysis (TGA) thermogram showing no mass loss before melting.

56. The crystalline form of any one of claims 2 and 49-55 having a differential thermal analysis (DTA) thermogram having a melting isotherm with an onset from about 189 °C to about 193 °C and a peak from about 193 °C to about 195 °C.

57. The crystalline form of any one of claims 2 and 49-56 having a DTA thermogram having a melting isotherm with an onset at about 191 °C and a peak at about 195 °C.

58. The crystalline form of any one of claims 2 and 49-57 having a TG/DTA thermogram substantially as depicted in Figure 35.

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59. The crystalline form of any one of claims 2 and 49-58 having a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting.

60. The crystalline form of any one of claims 2 and 49-58 having a melting endotherm at about 192 °C and a peak at about 196 °C.

61. The crystalline form of any one of claims 2 and 49-58 having a DSC thermogram substantially as depicted in Figure 36A or Figure 36B.

62. The crystalline form of any one of claims 2 and 49-61 having a proton nuclear magnetic resonance NMR) spectrum substantially as depicted in Figure 38.

63. The crystalline form of any one of claims 2 and 49-62 having a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 0.8% at 90% relative humidity during first sorption.

64. The crystalline form of any one of claims 2 and 49-63 having a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 37.

65. The crystalline form of claim 63 or 64, wherein the crystalline form of the HC1 salt is class II hygroscopic.

66. The crystalline form of any one of claims 2 and 49-65 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 776 cm^-1, 1693 cm^-1, 2349 cm^-1, 2933 cm^-1, 3062cm^-1, and 3177 cm^-1.

67. The crystalline form of any one of claims 2 and 49-66 having an FT-IR spectrogram substantially as depicted in Figure 39.

68. The crystalline form of claim 1, wherein the crystalline form of the HC1 salt is a monohydrate.

69. The crystalline form of claim 68 having an X-ray powder diffraction (XRPD) spectrum comprising at least 3 peaks with diffraction angles (29 ± 0.5°) selected from Table 4.

70. The crystalline form of claim 68 or 69 having an X-ray powder diffraction (XRPD) spectrum comprising at least 6 peaks with diffraction angles (29 ± 0.5°) selected from Table 4.

71. The crystalline form of any one of claims 68-70 having an X-ray powder diffraction (XRPD) spectrum comprising at least 10 peaks with diffraction angles (29 ± 0.5°) selected from Table 4.

72. The crystalline form of claim 68 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 15.1°, 17.4°, 19.8°, 20.0°, and 20.6°.

73. The crystalline form of claim 72, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 15.0°, 15.3°, 16.2°, 17.3°, and 26.0°.

74. The crystalline form of claim 68 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 15.1°, 17.4°, 19.8°, 20.0°, and 20.6°.

75. The crystalline form of claim 74, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 15.9°, 15.3°, 16.2°, 17.3°, and 26.9°.

76. The crystalline form of claim 68 having an XRPD spectrum substantially as depicted in Figure 12.

77. The crystalline form of any one of claims 72-76, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

78. The crystalline form of any one of claims 68-77 having a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 3.4 % from about 20 °C to about 80 °C.

79. The crystalline form of any one of claims 68-78 having a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 53 °C.

80. The crystalline form any one of claims 68-79 having a TG/DTA thermogram substantially as depicted in Figure 13.

81. The crystalline form of any one of claims 68-80 having a differential scanning calorimetry (DSC) thermogram having an endothermic peak with onset at about 70 °C and a peak at about 116 °C.

82. The crystalline form of any one of claims 68-81 having a DSC thermogram substantially as depicted in Figure 14A, Figure 14B, or Figure 14C.

83. The crystalline form of any one of claims 68-82 having a proton nuclear magnetic resonance (H NMR) spectrum substantially as depicted in Figure 15 A.

84. The crystalline form of any one of claims 68-83 having a heteronuclear single quantum correlation (HSQC) spectrum substantially as depicted in Figure 15B.

85. The crystalline form of any one of claims 68-84 having a gravimetric vapor sorption (GVS) isotherm plot substantially as depicted in Figure 16.

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86. The crystalline form of any one of claims 68-85 having a GVS kinetic plot substantially as depicted in Figure 17.

87. The crystalline form of any one of claims 68-86 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 764 cm^-1, 1657 cm^-1, 2977 cm^-1, 3202 cm^-1, and 3483 cm^-1.

88. The crystalline form of any one of claims 68-87 having an FT-IR spectrogram substantially as depicted in Figure 21.

89. The crystalline form of claim 1, wherein the crystalline form of the HC1 salt is a chloroform solvate.

90. The crystalline form of claim 89 having a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 17.6 % from about 20 °C to about 150 °C.

91. The crystalline form of claim 89 having a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 191 °C and a peak at about 195 °C.

92. The crystalline form of claim 1, wherein the crystalline form of the HC1 salt is an anisole solvate.

93. The crystalline form of claim 92 having a thermogravimetric analysis (TGA) thermogram having an initial mass loss of about 10.0 % from about 20 °C to about 130 °C.

94. The crystalline form of claim 92 having a differential thermal analysis (DTA) thermogram having an endothermic peak with onset at about 64 °C.

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95. A pharmaceutical composition comprising a crystalline form of a pharmaceutically acceptable salt of a compound of structure (1) according to any one of claims 1-94, wherein the crystalline form of a pharmaceutically acceptable salt of a compound of structure (1) is at least 98% pure.

96. The pharmaceutical composition of claim 95, wherein the crystalline form of a pharmaceutically acceptable salt of a compound of structure (1) is at least 99% pure.

97. A crystalline form of a compound having the following structure (1):

98. The crystalline form of claim 97 having an X-ray powder diffraction (XRPD) spectrum comprising at least 3 peaks with diffraction angles (29 ± 0.5°) selected from Table 2.

99. The crystalline form of claim 97 or 98 having an X-ray powder diffraction (XRPD) spectrum comprising at least 6 peaks with diffraction angles (29 ± 0.5°) selected from Table 2.

100. The crystalline form of any one of claims 97-99 having an X-ray powder diffraction (XRPD) spectrum comprising at least 10 peaks with diffraction angles (29 ± 0.5°) selected from Table 2.

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101. The crystalline form of claim 97 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.5°): 13.5°, 16.7°, 17.9°, 18.5°, and 19.8°.

102. The crystalline form of claim 101, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 12.3°, 18.4°, 19.5°, 21.2°, and 22.7°.

103. The crystalline form of claim 97 having an X-ray powder diffraction (XRPD) spectrum comprising peaks with the following diffraction angles (29 ± 0.2°): 13.5°, 16.7°, 17.9°, 18.5°, and 19.8°.

104. The crystalline form of claim 103, wherein the XRPD spectrum further comprises peak with the following diffraction angles (29 ± 0.5°): 12.3°, 18.4°, 19.5°, 21.2°, and 22.7°.

105. The crystalline form of claim 97 having an XRPD spectrum substantially as depicted in Figure 2.

106. The crystalline form of any one of claims 97-105, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

107. The crystalline form of any one of claims 97-106 having a differential scanning calorimetry (DSC) thermogram showing substantially no thermal events before melting.

108. The crystalline form of any one of claims 97-107 having a melting endotherm with a peak at about 188°C.

109. The crystalline form of any one of claims 97-108 having a DSC thermogram substantially as depicted in Figure 5.

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110. The crystalline form of any one of claims 97-109 having a proton nuclear magnetic resonance ( ’H NMR) spectrum substantially as depicted in Figure 4.

111. The crystalline form of any one of claims 97-110 having a dynamic vapor sorption (DVS) plot showing maximum mass increase of about 0.2% at 90% relative humidity during first sorption.

112. The crystalline form of any one of claims 97-111 having a dynamic vapor sorption (DVS) plot substantially as depicted in Figure 6.

113. The crystalline form of any one of claims 97-112, wherein the crystalline form of the free base is hygroscopic.

114. The crystalline form of any one of claims 97-113 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 1065 cm^-1, 2816 cm^-1, 2848 cm^-1, 2961 cm^-1, 2976 cm^-1, and 3085 cm^-1.

115. The crystalline form of any one of claims 97-114 having an FT-IR spectrogram substantially as depicted in Figure 7.

116. The crystalline form of claim 97 having a primitive orthorhombic lattice Bravais type.

117. The crystalline form of claim 116 having a space group of Pbca.

118. The crystalline form of claim 116, wherein the primitive orthorhombic lattice comprises vectors wherein a is about 10.4186 A, a is about 90°, b is about 11.6775 A, p is about 90°, c is about 35.442 A, and y is about 90°.

119. A crystalline form of a compound having the following structure (2):

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120. The crystalline form of claim 119 having an XRPD spectrum substantially as depicted in Figure 10.

121. A crystalline form of a napsylate salt of a compound having the following structure (1):

122. The crystalline form of claim 121 having an XRPD spectrum substantially as depicted in Figure 41.

123. The crystalline form of any one of claims 122, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

124. The crystalline form of any one of claims 121-123 having a TGA thermogram substantially as depicted in Figure 43.

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125. The crystalline form of any one of claims 121-124 having a proton nuclear magnetic resonance NMR) spectrum substantially as depicted in Figure 42.

126. The crystalline form of any one of claims 121-125 having a DSC thermogram substantially as depicted in Figure 44.

127. The crystalline form of any one of claims 121-126 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 622 cm^-1, 648 cm^-1, 867 cm^-1, 1394 cm^-1, 1663 cm^-1, and 2979 cm^-1.

128. The crystalline form of any one of claims 121-127 having an FT-IR spectrogram substantially as depicted in Figure 45.

129. A crystalline form of a xinafoate salt of a compound having the following structure (1):

130. The crystalline form of claim 129 having an XRPD spectrum substantially as depicted in Figure 46.

131. The crystalline form of claim 130, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

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132. The crystalline form of any one of claims 129-131 having a TGA thermogram substantially as depicted in Figure 48.

133. The crystalline form of any one of claims 129-132 having a proton nuclear magnetic resonance NMR) spectrum substantially as depicted in Figure 47.

134. The crystalline form of any one of claims 129-133 having a DSC thermogram substantially as depicted in Figure 49.

135. The crystalline form of any one of claims 129-134 having a Fourier- transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 532 cm^-1, 586 cm^-1, 659 cm^-1, 1494 cm^-1, 1657 cm^-1, and 2976 cm^-1.

136. The crystalline form of any one of claims 129-135 having an FT-IR spectrogram substantially as depicted in Figure 50.

137. The crystalline form of claim 129 having an XRPD spectrum substantially as depicted in Figure 51.

138. The crystalline form of claim 137, wherein the XRPD is measured using Cu K radiation in transmission mode using 40 kV / 40 mA generator settings.

139. The crystalline form of any one of claims 129 and 137-138 having a TGA thermogram substantially as depicted in Figure 53.

140. The crystalline form of any one of claims 129 and 137-139 having a proton nuclear magnetic resonance

NMR) spectrum substantially as depicted in Figure 52.

141. The crystalline form of any one of claims 129 and 137-140 having a DSC thermogram substantially as depicted in Figure 54.

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142. The crystalline form of any one of claims 129 and 137-141 having a Fourier-transform infrared (FT-IR) spectrogram comprising peaks with the following wavenumbers (± 4 cm^-1): 685 cm^-1, 876 cm^-1, 925 cm^-1, 1493 cm^-1, 1741 cm^-1, and 2983 cm^-1.

143. The crystalline form of any one of claims 129 and 137-142 having an FT- IR spectrogram substantially as depicted in Figure 55.

144. A pharmaceutical compositions comprising a crystalline form of any one of claims 1-143 and a pharmaceutically acceptable carrier.

145. The pharmaceutical composition of claim 144, wherein the crystalline form is 99% pure.

146. A method for inhibiting HIF-1α prolyl hydroxylase activity, comprising contacting the HIF-1α prolyl hydroxylase with an effective amount of a crystalline form of any one of claims 1-143 or a pharmaceutical composition of claim 144.

147. A method for treating a disease or condition for which HIF-1α prolyl hydroxylase inhibition is beneficial, comprising administering to a subject in need thereof an effective amount of a crystalline form of any one of claims 1-143 or a pharmaceutical composition of claim 144.

148. A method for treating an inflammatory disease, comprising administering to a subject in need thereof effective amount of a crystalline form of any one of claims 1-143 or a pharmaceutical composition of claim 144.

149. The method of claim 148, wherein the inflammatory disease is an inflammatory bowel disease.

150. The method of claim 149, wherein the inflammatory bowel disease is ulcerative colitis.

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151. The method of claim 149, wherein the inflammatory bowel disease is

Crohn’s disease.