WO2020222190A1 - Crystalline form of 6-[4-[1 -(propan-2-yl)piperidin-4-yl]-1,4-diazepan-1 -yl]-n-(pyrdin-4-yl)pyridine-2-carboxamide - Google Patents

Abstract

This invention relates to a novel crystalline form of a compound of Formula (I), which is 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide, and pharmaceutical compositions including the crystalline form. It also relates to the uses of such compositions, and to processes for the preparation of the crystalline forms and the compositions.

Description

CRYSTALLINE FORM OF 6-[4-[1 -(PROPAN-2-YL)PIPERIDIN-4-YL]-1 ,4-DIAZEPAN-1 -YL]-N-(PYRDIN-4-YL)PYRIDINE-2-CARBOXAMIDE

FIELD OF THE INVENTION This invention relates to a novel crystalline form of a compound of Formula (I), which is 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide, and pharmaceutical compositions including the crystalline form. It also relates to the uses of such compositions, and to processes for the preparation of the crystalline forms and the compositions.

BACKGROUND OF THE INVENTION

To formulate a pharmaceutically active compound into a suitably acceptable dosage form, it is desirable that the active compound possess acceptable stability and handling properties in addition to possessing acceptable biopharmaceutical properties such as solubility and dissolution. Certain crystalline, that is, morphological or polymorphic forms of compounds may be of interest to those involved in the development of suitable pharmaceutical dosage forms. If a certain polymorphic form is not held constant during clinical and stability studies, the exact dosage used or measured may not be comparable from one batch to the other. Once a pharmaceutical compound is produced for use, it is important to verify the morphological or polymorphic form delivered in each dosage form to assure that the production process delivers the same form and that the same amount of drug is included in each dosage. Therefore, it is imperative to assure that either a single morphological or polymorphic form or a known combination of morphological or polymorphic forms is present. In addition, certain morphological or polymorphic forms may exhibit enhanced thermodynamic stability and may be more suitable than other morphological or polymorphic forms for inclusion in pharmaceutical formulations.

To formulate a pharmaceutically active compound, such as 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide, into a suitably acceptable dosage form, it is desirable that the active compound possess acceptable stability and handling properties in addition to possessing acceptable biopharmaceutical properties such as solubility and dissolution.

It is therefore desirable to produce a crystalline form of 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide.

SUMMARY OF THE INVENTION

In one aspect, there is provided a various crystalline forms of 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide, the structure of which is as follows:

In particular, the crystalline form, or crystalline polymorph, is of Form 2 as defined below.

In a second aspect, there is provided a pharmaceutical composition comprising the crystalline polymorph of the first aspect of the invention.

In a third aspect, there provided a crystalline polymorph of the first aspect, or a pharmaceutical composition of the second aspect, for use in the treatment of a disease or condition responsive to reduction of CXCR4-mediated activity.

In a fourth aspect, there is a provided a process for preparing a crystalline polymorph.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows an exemplary synthetic pathway for 6-{4-[1 -(propan-2 -yl)piperidin- 4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide. How the compound is isolated from ethyl acetate can affect the resulting form of 6-{4-[1 - (propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide (for example, amorphous, Form 1 , or Form 2).

FIG. 2 shows an exemplary X-ray powder diffraction (XRPD) pattern for crystalline polymorph 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1.

FIG. 3 shows an exemplary XRPD pattern for crystalline polymorph 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2.

FIG. 4 shows an overlay of exemplary XRPD patterns for crystalline polymorph 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 1 (represented by the bottom line) and crystalline polymorph 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2 (represented by the top line).

FIG. 5 shows an exemplary differential scanning calorimetry (DSC) thermogram of crystalline polymorph 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1.

FIG. 6 shows an exemplary DSC thermogram of crystalline polymorph 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2.

FIG. 7 shows an exemplary thermogravimetric analysis (TGA)/DSC profile of crystalline polymorph 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1 and crystalline polymorph 6-{4-[1 - (Propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2.

FIG. 8 shows an overlay of exemplary Fourier Transform Infrared Spectroscopy (FT-IR) (4000 cm ¹ to 600 cm ¹) profiles of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1 and crystalline 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 2.

FIG. 9 shows an overlay of exemplary FT-IR (3500 cm ¹ to 1800 cm ¹) profiles of crystalline 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide Form 1 and crystalline 6-{4-[1 -(propan-2 -yl)piperidin- 4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 2.

FIG. 10 shows an overlay of exemplary FT-IR analysis (1800 cm ¹ to 600 cm ¹) profiles of crystalline 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1 (solid line) and crystalline 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2 (dashed line).

FIG. 1 1 shows a representative image of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base Form 1.

FIG. 12 shows a representative image of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base Form 2.

FIG. 13 shows an exemplary dynamic vapor sorption (DVS) study sorption/desorption isotherm of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan- 1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1.

FIG 14 shows a conformational picture of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 , 4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1 based on single crystal X-ray analysis. DETAILED DESCRIPTION OF THE INVENTION

This disclosure describes novel crystalline forms of a compound of Formula (I). The compound of Formula (I) is 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan- 1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide.

Anywhere in the present application where a name of a compound may not correctly describe the structure of the compound, the structure supersedes the name and governs.

6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide has been shown to be an antagonist of the CXCR4 receptor, and could be useful in the treatment of conditions which respond to antagonism of the CXCR4 receptor, such as cancer, HIV/AIDS, neuropathy, HIV related neuropathy, pain, inflammation, brain inflammation, neurodegeration, cognitive degeneration, diabetic retinopathy, age related macular degeneration, retinal neo-vascularisation, and viral infections. The cancer may be a CNS cancer, and in particular glioblastoma or astrocytoma, especially glioblastoma.

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide may be administered to a patient in combination with one or more chemotherapeutic agents, including brain penetrating chemotherapeutic agents, such as a DNA modifying agent, optionally in combination with irradiation. Suitable regimes of irradiation and examples of chemotherapeutic agents can be found in the current guidelines: 201 1 Canada, Easaw et al. , Current Oncology Vol 18 No 3., which is incorporated herein by reference.

The terms "treat" or "treatment" refer to therapeutic or palliative measures. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (that is, not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment.

The term "a therapeutically effective amount" or "effective amount" means an amount of the compound sufficient to induce a therapeutic or prophylactic effect. The exact amount of active compound used in a pharmaceutical composition of the invention will vary according to factors known to those of skill in the art, such as the physical and chemical nature of the compound, the nature of the carrier, and the intended dosing regimen.

The term "solvate" is used herein to describe a molecular complex comprising the compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term ‘hydrate’ is employed when said solvent is water.

The words "preferred" and "preferably" refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.

As used herein, the term“stable” means that no degradation (for example, less than 0.01 %, less than 0.1 %, less than 0.5% by weight) occurs during the specified time period and specified conditions (such as for at least 3 months under accelerated conditions (for example, 40°C and 75% RH), for at least 6 months under accelerated conditions, or for at least 9 months under accelerated conditions).

The terms "comprises" and variations thereof do not have a limiting meaning where these terms appear in the description and claims. Unless otherwise specified, "a," "an," "the," and "at least one" are used interchangeably and mean one or more than one.

Unless otherwise indicated, all numbers expressing quantities of components, molecular weights, and so forth used in the specification and claims are to be understood as being modified in all instances by the term“about”. Accordingly, unless otherwise indicated to the contrary, the numerical parameters set forth in the specification and claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention.

Also herein, the recitations of numerical ranges by endpoints include all numbers subsumed within that range (for example, 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

As used herein, the recitation of a numerical range for a variable is intended to convey that the invention may be practiced with the variable equal to any of the values within that range. Thus, for a variable that is inherently discrete, the variable can be equal to any integer value of the numerical range, including the end-points of the range. Similarly, for a variable that is inherently continuous, the variable can be equal to any real value of the numerical range, including the end-points of the range. As an example, a variable that is described as having values between 0 and 2 or in a range of 0 to 2, can be 0, 1 , or 2 for variables that are inherently discrete, and can be 0.0, 0.1 , 0.01 , 0.001 , or any other real value for variables that are inherently continuous.

The term“about” is used herein to mean approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In some embodiments, the term "about" is used herein to modify a numerical value above and below the stated value by a variance of 20%, preferably 10%. With respect to peak positions in an XRPD pattern, the term“about” means that the peak, as reported in terms of angular positions (two theta), has an allowable variability of ± 0.3°. The variability of ± 0.3° is intended to be used when comparing two XRPD patterns. In practice, if a diffraction pattern peak from one pattern is assigned a range of angular positions (two theta) which is the measured peak position ± 0.3° and if those ranges of peak positions overlap, then the two peaks are considered to have the same angular position. For example, if a peak from one pattern is determined to have a position of 11.0°, for comparison purposes the allowable variability allows the peak to be assigned a position in the range of 10.7°-1 1.3°.

The term“crystalline” is used herein as a synonym for, and interchangeably with, the phrases “crystalline compound,” “crystalline form,” “polymorphic form,” “morphological form” and the like.

The terms "polymorph" and "polymorphic form" refer to different crystalline forms of a single compound. That is, polymorphs are distinct solids sharing the same molecular formula, yet each polymorph may have distinct solid state physical properties. Therefore, a single compound may give rise to a variety of polymorphic forms where each form has different and distinct solid state physical properties, such as different solubility profiles, dissolution rates, melting point temperatures, flowability, and/or different X-ray diffraction peaks. The differences in physical properties may affect pharmaceutical parameters such as storage stability, compressibility and density (which can be important in formulation and product manufacturing), and dissolution rate (which can be an important factor in bioavailability). Techniques for characterizing polymorphic forms include, but are not limited to, X-ray powder diffractometry (XRPD), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), single-crystal X-ray diffractometry (XRD), vibrational spectroscopy, for example, infrared (IR) and Raman spectroscopy, solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility measurements, dissolution measurements, elemental analysis and Karl Fischer analysis. In a first aspect of the invention, there is provided a crystalline polymorph comprising a compound of Formula (I). The crystalline polymorph may consist of a compound of Formula (I).

Polymorphic forms of any given compound may be distinguished from each other using different characterization or identification techniques. For example, conventional organic chemistry identification techniques may be used to distinguish different polymorphic forms. Such identification techniques may include, but are not limited to: X-Ray Powder Diffraction (XRPD), Single Crystal X-Ray Diffraction (SCXRD), Nuclear Magnetic Resonance (NMR) (that is, such as Proton Magnetic Resonance (¹H NMR), ¹³C Nuclear Magnetic Resonance (¹³C NMR), Infrared Spectroscopy (IR), Fourier Transform Infrared Spectroscopy (FT-IR), Thermo-Gravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC), Thermo-Gravimetric Analysis (TGA), Karl Fisher Titration (KFT), Gravimetric Vapor Sorption (GVS), Electron Microscopy, Polarized Light Microscopy (PLM), Hot Stage Microscopy (HSM), Optical Crystallography, Dilatometry, etc.

The compound of Formula (I) may be the free base of 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide, or a pharmaceutically acceptable salt thereof. It is preferred that the compound is the free base.

The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a mammal such as a human (for example, do not produce an adverse, allergic or other unwanted reaction when administered to a mammal).

As used herein the term “salt” includes base addition, acid addition and ammonium salts. 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide is basic and so can form salts, including pharmaceutically acceptable salts with inorganic acids, e.g. with hydrohalic acids such as hydrochloric or hydrobromic acids, sulphuric acid, nitric acid or phosphoric acid and the like, and with organic acids e.g. with acetic, trifluoroacetic, tartaric, succinic, fumaric, maleic, malic, salicylic, citric, methanesulphonic, p-toluenesulphonic, benzoic, benzenesulfonic, glutamic, lactic, and mandelic acids and the like. Those compounds which have a basic nitrogen can also form quaternary ammonium salts with a pharmaceutically acceptable counter-ion such as chloride, bromide, acetate, formate, p- toluenesulfonate, succinate, hemi-succinate, naphthalene-bis sulfonate, methanesulfonate, trifluoroacetate, xinafoate, and the like. For a review on salts, see “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002), which is incorporated herein by reference.

A feature of the first aspect of the invention is that the compound of Formula (I) is crystalline Form 2, subsequently referred to as“Form 2”.

Form 2 may be characterized in several different ways. It may exhibit an X-ray powder diffraction (XRPD) pattern with peaks at 8.9, 15.9, 18.6, and 21.8°2Q. It may exhibit an XRPD pattern with peaks at 8. 2Q, 8.9°2Q, 9.9°2Q, 14.4°2Q, 15.9°2Q, 16.8°2Q, 18.6°2Q, 20.2°2Q, 21.8°2Q, and 25.4°2Q. These peaks may be at 8.9°2Q ± 0.3°2Q, 15.9°2Q ± 0.3°2Q, 18.6°2Q ± 0.3°2Q, and 21.8°2Q ± 0.3°2Q, and may further comprise at least one peak appearing at 8.1 °2Q ± 0.3°2Q, 9.9°2Q ± 0.3°2Q, 14.4°2Q ± 0.3°2Q, 16.8°2Q ± 0.3°2Q, 20.2°2Q ± 0.3°2Q, or 25.4°2Q ± 0.3°2Q. A particular feature of the invention is that the XRPD pattern comprises at least 3, at least 4, at least 5, at least 6, at least 8, or at least 10, of the preceding peaks. Form 2 may exhibit an XRPD pattern that is substantially the same as the XRPD pattern of FIG. 3.

The term "substantially the same as" as used herein, refers to a XRPD pattern, differential scanning calorimetry pattern, or Fourier transform infrared spectrophotometry pattern that is non-identical to those depicted herein, but that falls within the limits of experimental error, when considered by one of ordinary skill in the art. Form 2 may have a DSC thermogram exhibiting an endothermic event having a melt maxima temperature in the range of 157 °C to 159 °C. Form 2 may have a DSC thermogram substantially the same as the DSC thermogram of FIG. 6.

Form 2 may exhibit Fourier-transform infrared (FTIR) spectrophotometry substantially the same as the Form 2 FTIR spectrum of at least one of FIG. 8, FIG. 9, and FIG. 10.

The compound of Formula (I) may also exist in crystalline Form 1 , subsequently referred to as“Form 1”.

Form 1 may be characterised in several different ways. It may exhibit an XRPD pattern with peaks at 8.9°2Q, 13.4°2Q, 17.9°2Q, and 22.4°2Q. It may exhibit an XRPD pattern with peaks at 8.9°2Q, 12.1 °2Q, 13.4°2Q, 16.2°2Q, 17.4°2Q, 17.9°2Q, 22.4°2Q, 23.6°2Q, 26.6°2Q, 28.2°2Q, 29.9°2Q, 31.6°2Q, and 36.2°2Q. These peaks may be at 8.9°2Q ± 0.3°2Q, 13.4°2Q ± 0.3°2Q, 17.9°2Q ± 0.3°2Q, and 22.4°2Q ± 0.3°2Q, and may further comprise at least one peak appearing at 12.1 °2Q ± 0.3°2Q, 16.2°2Q ± 0.3°2Q, 17.4°2Q ± 0.3°2Q, 23.6°2Q ± 0.3°2Q, 26.6°2Q ± 0.3°2Q, 28.2°2Q ± 0.3°2Q, 29.9°2Q ± 0.3°2Q, 31.6°2Q ± 0.3°2Q, or 36.2°2Q ± 0.3°2Q. The XRPD pattern may comprises at least 3, at least 4, at least 5, at least 6, at least 8, or at least 10, of the preceding peaks. Form 1 may have an XRPD peak at 16.2°2Q ± 0.3°2Q. Form 1 may exhibit an XRPD pattern that is substantially the same as the XRPD pattern of FIG. 2.

Form 1 may have a differential scanning calorimetry (DSC) thermogram exhibiting an endothermic event having a melt maxima temperature in the range of 141 °C to 142 °C. Form 1 may have a DSC thermogram substantially the same as the DSC thermogram of FIG. 5.

Form 1 may exhibit Fourier-transform infrared (FTIR) spectrophotometry substantially the same as the Form 1 FTIR spectrum of at least one of FIG. 8, FIG. 9, and FIG. 10. The crystalline polymorph of Form 1 may be a single crystal form of 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide having an orthorhombic crystal system, a Pbca space group, and the following unit cell dimensions: a = 12.4413(5) Angstroms (A), b = 8.9899 (5) A, c = 39.343 (2) A, a = 90°, b = 90°, g = 90°.

Form 1 may exhibit a single crystal data as disclosed in Example 1 1 and/or FIG. 14.

When the crystalline polymorph of the invention comprises Form 2, it may optionally comprise (or be intermixed or combined with) a certain amount of another polymorph, such as Form 1 , or amorphous 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide. The crystalline polymorph of the invention may comprise less than about 50 weight percent, such as less than about 25 wt%, for instance less than about 15 wt%, preferably less than about 10 wt%, more preferably less than about 5 wt%, of Form 2 and/or amorphous 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}- N-(pyridin-4-yl)pyridine-2-carboxamide.

In a second aspect of the invention, there is provided a pharmaceutical composition comprising the crystalline polymorph of the first aspect of the invention, and a pharmaceutically acceptable carrier and/or excipient. It is preferable that the pharmaceutical composition comprises Form 2, but it may also comprise Form 1.

The pharmaceutical composition may be prepared by a method that includes combining a pharmaceutically acceptable carrier and the crystalline polymorph of Form 2, optionally in combination with crystalline polymorph Form 1 , in an amount effective to provide a therapeutically effective amount of the compound of Formula (I), i.e. 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide.

For clinical use, the crystalline polymorph of the invention may be formulated into pharmaceutical formulations for various modes of administration. It will be appreciated that the crystalline polymorph of the invention may be administered together with a physiologically acceptable carrier, excipient, or diluent. The pharmaceutical compositions of the invention may be administered by any suitable route, preferably by oral, rectal, nasal, topical (including buccal and sublingual), sublingual, transdermal, intrathecal, transmucosal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration.

Formulations may conveniently be presented in unit dosage form, for example, tablets and sustained release capsules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy. Pharmaceutical formulations are usually prepared by mixing the active substance, or a pharmaceutically acceptable salt thereof, with conventional pharmaceutically acceptable carriers, diluents or excipients. Examples of excipients are water, gelatin, gum arabicum, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talcum, colloidal silicon dioxide, and the like. Such formulations may also contain other pharmacologically active agents, and conventional additives, such as stabilizers, wetting agents, emulsifiers, flavouring agents, buffers, and the like. Usually, the amount of active compounds is between 0.1 percent (%) and 95% by weight of the preparation, preferably between 0.2 % and 20% by weight in preparations for parenteral use and more preferably between 1 % and 50% by weight in preparations for oral administration.

The formulations can be further prepared by known methods such as granulation, compression, microencapsulation, spray coating, etc. The formulations may be prepared by conventional methods in the dosage form of tablets, capsules, granules, powders, syrups, suspensions, suppositories or injections. Liquid formulations may be prepared by dissolving or suspending the active substance in water or other suitable vehicles. Tablets and granules may be coated in a conventional manner. To maintain therapeutically effective plasma concentrations for extended periods of time, the compound of the invention may be incorporated into slow release formulations. The dose level and frequency of dosage of the specific compound will vary depending on a variety of factors including the potency of the specific compound employed, the metabolic stability and length of action of that compound, the patient's age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the condition to be treated, and the patient undergoing therapy. The daily dosage may, for example, range from 0.001 milligram (mg) to 100 mg per kilogram (kilo) of body weight, administered singly or multiply in doses, for example, from 0.01 mg to 25 mg each. A typical total daily dosage for a human is 1 milligram per day (mg/day) to 2000 mg/day, preferably from 200 mg/day to 2000 mg/day, more preferably from 500 mg/day to 2000 mg/day. Normally, such a dosage is given orally but parenteral administration may also be chosen.

The compound of Formula (I) has CXCR4 inhibitory activity. The crystalline polymorph, or pharmaceutical composition comprising the crystalline polymorph, of the invention may therefore be particularly useful as a medicament, for instance in the treatment of a disease or condition responsive to reduction of CXCR4-mediated activity.

In a third aspect of the invention there is therefore provided a crystalline polymorph of the first aspect of the invention, or a pharmaceutical composition of the second aspect of the invention, for use in the treatment of a disease or condition responsive to reduction of CXCR4-mediated activity.

The crystalline polymorph of the first aspect of the invention may also be used in the manufacture of a medicament for use in the treatment of a disease or condition responsive to reduction of CXCR4-mediated activity.

There is also provided a method for treating a subject having a disease or condition responsive to reduction of CXCR4-mediated activity, the method comprising administering an effective amount of a crystalline polymorph of the first aspect of the invention, or a pharmaceutical composition of the second aspect of the invention, to a subject in need thereof. The disease or condition may be selected from the group consisting of cancer, HIV/AIDS, neuropathy, HIV related neuropathy, pain, inflammation, brain inflammation, neurodegeration, cognitive degeneration, diabetic retinopathy, age related macular degeneration, retinal neo-vascularisation, and viral infections.

The crystalline polymorph or pharmaceutical composition may be particularly useful in the treatment of cancer. The cancer may be selected from the group consisting of cancer of the breast, lung (including non-small cell and small cell), pancreas, cervix, thyroid, kidney, bladder, ovary, prostate, skin (including melanoma), cancer of the Gl tract (including oesophageal, hepatic, colorectal and gastric cancers), oral squamous carcinoma, cancers of the blood including leukaemias such as B-CLL, AML, CML, ALL, lymphomas such as intraocular, non-Hodgkins and Hodgkins lymphomas, and multiple myeloma; neuroblastoma, cancers of the nervous system including cancer of the brain, glioma, glioblastoma, other astrocytomas, oligodendroglial tumour, meningioma, ependymoma, oligodendroglioma, medulloblastoma, and metastases into the CNS from peripheral cancers. They are particularly useful in the treatment of CNS cancers, for instance wherein the CNS cancer is cancer of the brain. The CNS cancer may be a glioma. The CNS cancer may be selected from the group consisting of glioblastoma, other astrocytomas, oligodendroglial tumour, meningioma, ependymoma, oligodendroglioma, medulloblastoma, and metastases into the CNS from peripheral cancers. The CNS cancer is preferably selected from glioblastoma and astrocytoma, especially glioblastoma.

In the above diseases and conditions the compound of Formula (I) may be administered in combination with one or more CHK1 inhibitors, and/or one or more anti-angiogenic agents, preferably with one or more CHK1 inhibitors.

The anti-angiogenic agent may be an inhibitor of the VEGF receptor or an inhibitor of the PDGF receptor. The anti-angiogenic agent may be selected from the group consisting of cediranib, sunitinib, sorafenib, pazopanib, tivozanib vatalanib, vandertanib, brivanib, dovitinib, motesanib, telatinib and axitinib. The treatment may comprise an apheresis procedure for promoting release and mobilisation of stem cells, including haematopoietic and non-haematopoietic stem cells and progenitor stem cells prior to harvesting. In such cases, GCSF may be used in the apheresis procedure and/or the apheresis procedure may be implemented prior to treatment of a subject by chemotherapy or radiotherapy, for reducing chemotherapy- or radiotherapy-induced leukopenia.

The crystalline polymorph or pharmaceutical composition of the invention may be administered in a variety of dosage forms. Thus, they can be administered orally, for example as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules. The crystalline polymorph or pharmaceutical composition may be administered in a sublingual formulation, for example a buccal formulation. The crystalline polymorph or pharmaceutical composition of the invention may also be administered parenterally, whether subcutaneously, intravenously, intramuscularly, intrasternally, transdermally, by inhalation, intranasally, or by infusion techniques. The crystalline polymorph or pharmaceutical composition may also be administered as suppositories. Thus, the crystalline polymorph or pharmaceutical composition of the invention may be administered orally, or by inhalation, or intranasally, but preferably the crystalline polymorph or pharmaceutical composition of the invention may be administered orally and more preferably, the crystalline polymorph or pharmaceutical composition of the invention may be administered as a tablet or capsule. In the latter connection, administration of the crystalline polymorph or pharmaceutical composition in a hard gelatin capsule form, or in one of the many sustained release formulations known in the art will often be preferred.

The crystalline polymorph of the invention may typically be formulated for administration with a pharmaceutically acceptable carrier or diluent. For example, solid oral forms may contain, together with the active compound, diluents, for example, lactose, dextrose, saccharose, cellulose, corn starch or potato starch; lubricants, for example, silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glycols; binding agents; for example, starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents, for example, starch, alginic acid, alginates or sodium starch glycolate; effervescing mixtures; dyestuffs; sweeteners; wetting agents, such as lecithin, polysorbates, laurylsulphates; and, in general, non-toxic and pharmacologically inactive substances used in pharmaceutical formulations. Such pharmaceutical preparations may be manufactured in known manner, for example, by means of mixing, granulating, tableting, sugar coating, or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions and suspensions. The syrups may contain as carriers, for example, saccharose or saccharose with glycerine and/or mannitol and/or sorbitol. Suspensions and emulsions may contain as carrier, for example a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol. The suspension or solutions for intramuscular injections may contain, together with the active compound, a pharmaceutically acceptable carrier, for example, sterile water, olive oil, ethyl oleate, glycols, for example, propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.

It will be understood that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing treatment. Optimum dose levels and frequency of dosing will be determined by clinical trial, as is required in the art. However, it is expected that a typical dose will be in the range from 0.001 to 50 mg per kg of body weight.

The useful treatments include those described in WO 2012/049277 and/or WO 2016/157149, both of which are incorporated herein by reference.

In a fourth aspect of the invention there is provided a process for preparing a crystalline polymorph according to the first aspect of the invention.

Amorphous 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide, i.e. a compound of Formula (I), may be prepared as described below and as described in Example 30 of WO 2012/049277, which is incorporated herein by reference.

6-Chloropyridine-2-carboxylic acid (5.00 g, 31.7 mmol) was dissolved in dichloromethane (DCM) (150 ml_) and oxalyl chloride (5.45 ml_, 63.5 mmol) and A/,/V-dimethylformamide (DMF) (1 ml_) were added. The reaction mixture was stirred for 3 hours, concentrated in vacuo and azeotroped with DCM. The residue was dissolved in tetrahydrofuran (THF) (150 ml_) and potassium tert- butoxide (3.39 mg, 47.6 mmol) was added. The reaction mixture was stirred for 18 hours, quenched with water (250 ml_) and extracted with DCM (3 ^c 150 ml_). The combined organic fractions were washed with saturated aqueous (sat aq) NaHCOs (150 ml_), dried (MgS0₄) and concentrated in vacuo. The residue was purified by column chromatography to give tert-Butyl 6-chloropyridine-2- carboxylate (3.1 1 g, 46%) as a white solid. Liquid Chromatography Mass Spectrometry (LCMS) Electrospray (ES⁺): 236.1 [MNa]⁺.

tert-Butyl 6-chloropyridine-2-carboxylate was dissolved in DMA (60 mL) and homopiperazine (7.03 g, 70.2 mmol) was added. The reaction mixture was heated using a microwave (180 °C, absorption high) for 35 minutes and the solvents were removed in vacuo. The residue was dissolved in DCM (150 mL) and washed with sat aq Na₂C0₃(100 mL), brine (100 mL), dried (MgS0₄) and the solvents were removed in vacuo to give crude tert-butyl 6-(1 ,4-diazepan-1 - yl)pyridine-2-carboxylate (3.37 g, 87%) as a yellow liquid. LCMS (ES⁺): 278.1 [MH]⁺.

tert-Butyl 6-(1 ,4-diazepan-1 -yl)pyridine-2-carboxylate (3.37 g, 12.2 mmol) was dissolved in DCM (125 mL) and 1 -(propan-2-yl)piperidin-4-one (3.61 mL, 24.3 mmol) and NaBH(OAc)₃ (12.9 g, 60.8 mmol) were added. The reaction mixture was stirred for 18 hours, diluted with DCM (250 mL) and quenched with saturated aqueous Na₂C0₃ (150 mL). The aqueous fraction was extracted with DCM (2 x 100 mL) and the combined organic fractions were dried (MgSC ) and concentrated in vacuo. The residue was purified by column chromatography to give tert-Butyl 6-{4-[1 -(propan-2- l)piperidin-4-yl]-1 ,4-diazepan-1 -yl}pyridine-2- carboxylate (3.29 g, 67%) as a yellow liquid. LCMS (ES⁺): 403.5 [MH]⁺.

tert-Butyl 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}pyridine-2- carboxylate was dissolved in DCM (80 mL), TFA (40 mL) was added and the reaction mixture was stirred for 18 hours. The solvents were removed in vacuo and the residue was neutralized with 1 M aq Na₂C0₃. The aqueous solution was washed with DCM, concentrated in vacuo and purified by reverse phase column chromatography to give 6-{4-[1 -(propan-2 -yl)piperidin-4- l]-1 ,4-diazepan-1 - yl}pyridine-2-carboxylic acid (2.19 g, 77%) as a white solid. LCMS (ES⁺): 347.5 [MH]⁺.

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}pyridine-2-carboxylic acid (650 mg, 1.88 mmol), 4-aminotetrahydropyran (210 mg, 2.07 mmol), N,N- diisopropylethylamine (DIPEA) (970 mg, 7.52 mmol) and 2-(1 H-Benzotriazole-1 - yl)-1 , 1 ,3,3-tetramethyluronium hexafluorophosphate (HBTU) (710 mg, 1.88 mmol) were dissolved in DMF (10 mL) and the reaction mixture was stirred for 20 hours. The solvents were removed in vacuo and the residue was diluted with DCM (100 mL), washed with sat aq Na₂COs (10 mL), dried (MgS0₄) and the solvents were removed in vacuo. The residue was purified by reverse phase high performance liquid chromatography (HPLC) and de-salted (K₂C0₃ in DCM) to give 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide (300 mg, 38%) as a colorless gum. HRMS (ESI+) calculated for C₂₄H₃₄N₆0: 422.27941 , found 422.27983. HPLC: R_f 3.18 min, 100% purity.

Amorphous compound of Formula (I) may be prepared as described in Figure 1 and/or Example 1.

The process for preparing amorphous compound of Formula (I) may include heating the compound to a molten state, and then cooling rapidly to solidify. The compound may be heated to about 170 °C. The material can be cooled at a rate of 200 °C per minute.

The process of the fourth aspect of the invention for preparing a crystalline polymorph may comprise the steps of

(i’) mixing the compound of Formula (I) with a suitable solvent to form a first mixture;

(ii’) heating the first mixture to form a second mixture;

(iii’) cooling the second mixture to form a third mixture; and

(iv’) isolating the crystalline polymorph.

This method is particularly useful in the formation of Form 2.

In step (i’), the compound of Formula (I) may be mixed with the suitable solvent by any suitable means, such as using a paddle mixer or an overhead mixer.

The suitable solvent may be a polar or non-polar solvent. Such solvents include, but are not limited to water, DMSO, ethylene glycol, DMF, NMP, MIBK, 1 ,2- dimethoxyethane, IPAc, 2-ethoxyethanol, 1 ,2-xylene, toluene, 1 ,4-dioxane, and heptane. A polar solvent may be preferred because the kinetics of forming Form 2 may be enhanced. The solvent preferably includes water, if slurrying at room temperature is involved in step (iv’). For instance the solvent may comprise water and ethyl acetate, water and methyl ethyl ketone (MEK), or water and isopropyl alcohol (I PA). The solvent may preferably include methyl ethyl ketone.

A slurry may be a mixture comprising a solid and a solvent. In a slurry, the solid may be only sparingly soluble in the solvent.

Where the solvent includes water and another compound, the solvent may include at least 1.0 volume/volume percent (% v/v) and up to 2.0 % v/v water. The solution is preferably 1.5 % v/v water. The solvent may include at least 10 volumes, at least 12 volumes, at least 15 volumes and up to 15 volumes, or up to 17 volumes of ethyl acetate, MEK or I PA. The solution preferably includes 10 volumes of ethyl acetate, MEK or I PA.

The first mixture may be heated by any conventional means, such as in an oven or the like. When the first mixture is heated in step (ii’) (i.e. to form the second mixture) the compound of Formula (I) may be dissolved to form a solution, or it may remain, or remain in part, as a solid in which case a slurry may be formed.

Form 1 and Form 2 of the compound of Formula (I) are enantiotropically related. Form 1 may be stable at low temperatures and Form 2 may be stable at high temperatures. The calculated and observed polymorphic transition temperature between Form 1 and Form 2 is about 69 °C.

In step (ii’), the first mixture may be heated to a temperature of greater than about 69 °C, such as from about 75 °C to about 150 °C, preferably to a temperature of from about 80 °C to about 120 °C, more preferably from about 85 °C to about 1 10 °C, even more preferably from about 90 °C to about 100 °C, to form the second mixture. It will be understood that the suitable solvent may be heated to the desired temperature prior to the compound being mixed with the solvent. The second mixture may be held at that temperature for a suitable time period, which may be for at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 24 hours. It is preferable that the second mixture, when heated, is a slurry. If a slurry is used in step (ii’), the slurry may be slurried for at least 1 hour, at least 3 hours, at least 6 hours, at least 12 hours, at least 18 hours, at least 24 hours, at least 36 hours, at least 48 hours, at least 72 hours, or at least 120 hours. In some embodiments, the slurry can be slurried for up to 24 hours, up to 48 hours, up to 72 hours, up to 120 hours, or up to 168 hours. The process may include centrifuge filtration of the Form 2 produced during the slurrying.

In step (iii’) of the process, the second mixture is allowed to cool to form the third mixture. The second mixture may be cooled rapidly. The solvent may be cooled to a temperature in a range of 0 °C to 5 °C. This may be achieved using an ice bath. The solvent may be cooled over a period of up to 5 minutes, up to 10 minutes, up to 15 minutes, or up to 20 minutes. After cooling, the mixture may be stirred at a temperature in a range of 0 °C to 5 °C for at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, or at least 3 hours.

In step (iv’), the crystalline polymorph may be isolated by centrifuge vacuum filtration and displacement, or by vacuum filtration. Form 2 may be washed, for instance with 1 volume of solvent. The solvent may be at a temperature in a range of 0 °C to 5 °C. Form 2 may then be dried including, for example, in a vacuum oven.

Form 2 can be prepared as described in Example 3-B, Example 4 and Example 13.

Form 1 may be produced by a method analogous to Form 2, i.e. a process that comprises the steps of

(i) mixing the compound of Formula (I) with a suitable solvent to form a first mixture;

(ii) heating the first mixture to form a second mixture;

(iii) cooling the second mixture to form a third mixture; and

(iv) isolating the crystalline polymorph. In step (i), the compound of Formula (I) may be mixed with the suitable solvent by any suitable means, such as using a paddle mixer or an overhead mixer.

The suitable solvent may be a polar or non-polar solvent. Such solvents include, but are not limited to, DMSO, ethylene glycol, DMF, acetonitrile, ethanol, acetone, IPA, MEK, MIBK, 1 ,2-dimethoxyethane, methyl-THF, methyl acetate, ethyl acetate, IPAc, diethyl ether, MTBE, 1 ,2-xylene, toluene, 1 ,4-dioxane, and heptane. The solvent preferably includes at least one of ethyl acetate, methyl ethyl ketone, and isopropyl alcohol. In some embodiments, the solvent includes water.

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide may be mixed with a solvent including water and ethyl acetate. In some embodiments, including, for example, where the solvent includes water and ethyl acetate, the solvent includes at least 1.0 volume/volume percent (% v/v) and up to 2.0 % v/v water. In some embodiments, the solvent is preferably a 1.5 % v/v water in ethyl acetate solution. In some embodiments, the solvent includes at least 10 volumes, at least 12 volumes, at least 15 volumes and up to 15 volumes, or up to 17 volumes of ethyl acetate. In some embodiments, the solvent preferably includes 12 volumes of ethyl acetate.

The first mixture may be heated by any conventional means, such as in an oven or the like. When the first mixture is heated in step (ii) (i.e. to form the second mixture) the compound of Formula (I) may be dissolved to form a solution, or it may remain, or remain in part, as a solid in which case a slurry may be formed.

In a feature of the fourth aspect of the invention, in step (ii), the first mixture is heated to a temperature of about 69 °C or less, such as from about 45 °C to about 69 °C, preferably to a temperature of from about 50 °C to about 68 °C, more preferably from about 53 °C to about 68 °C, to form the second mixture. It will be understood that the suitable solvent may be heated to the desired temperature prior to the compound being mixed with the solvent. The second mixture may be held at the desired temperature for a suitable time period, which may be for at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 24 hours.

In step (iii) of the process, the second mixture is allowed to cool to form the third mixture. The mixture may be transferred to a crystallisation vessel for the cooling process. The transfer may be facilitated via an in-line filter.

The second mixture may be cooled to a temperature of about 40 °C or less, such as about 35 °C or less, for instance 30°C or less, preferably 25 °C or less, more preferably 20 °C or less, and most preferably 15°C or less. When the mixture is cooled to a temperature in a range of 0°C to 5°C, the mixture may be cooled over at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, or at least 8 hours. When the mixture is cooled to a temperature in a range of 0°C to 5°C, the mixture is then held at a temperature in a range of 0°C to 5°C for at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 60 minutes, or at least 2 hours.

If the second mixture is a solution when heated to the desired temperature in step (ii), the cooling step (iii) may form the compound of Formula (I) as Form 1 , i.e. during the cooling step, nucleation and/or precipitation of the compound of Formula (I) in Form 1 may occur. In this case, the third mixture may be slurried prior to isolation of the crystalline polymorph. During or before the slurrying process, a portion of the suitable solvent may be removed from the mixture. The suitable solvent may be removed by evaporation or distillation including, for example, distillation under reduced pressure. The volume of the mixture may be reduced by at least 40%, at least 50%, at least 60%, at least 70, at least 80%, at least 90%, or at least 95%, by volume compared to the volume prior to solvent removal. The mixture may be slurried before a portion of the solvent is removed. Additional nucleation and/or precipitation of the compound of Formula (I) in Form 1 may occur during slurrying. The mixture may be slurried at a temperature of about 25 °C or less, such as about 23 °C or less, for instance about 22 °C or less, preferably about 20 °C or less, more preferably about 15 °C or less. The mixture may be slurried at ambient temperature. The mixture may be slurried for at least about 1 hour, for instance at least about 3 hours, for example at least about 6 hours, at least about 12 hours, at least about 18 hours, at least about 24 hours, at least about 36 hours, at least about 48 hours, at least about 72 hours, or at least about 120 hours.

If the second mixture is a slurry when heated to the desired temperature in step (ii), the compound of Formula (I) as Form 1 may be formed by slurrying the second mixture at that desired temperature, i.e. prior to cooling the second mixture in step (iii). In this case, the second mixture may be slurried at a temperature of about 69 °C or less, such as from about 45 °C to about 69 °C, preferably from about 50 °C to about 68 °C, more preferably from about 53 °C to about 68 °C. During or before the slurrying process, a portion of the suitable solvent may be removed from the mixture. The suitable solvent may be removed by evaporation or distillation including, for example, distillation under reduced pressure. The volume of the mixture may be reduced by at least 40%, at least 50%, at least 60%, at least 70, at least 80%, at least 90%, or at least 95%, by volume compared to the volume prior to solvent removal. The mixture may be slurried for at least about 1 hour, for instance at least about 3 hours, for example at least about 6 hours, at least about 12 hours, at least about 18 hours, at least about 24 hours, at least about 36 hours, at least about 48 hours, at least about 72 hours, or at least about 120 hours.

In step (iv), the crystalline polymorph may be isolated by centrifuge filtration. The crystalline polymorph may be dried. For example, the crystalline polymorph may be transferred to a filter and dried on the filter. The crystalline polymorph may be dried on the filter for at least 6 hours. Other forms of drying may include, for example, tray drying, fluid bed drying, vacuum drying, rotary drum drying, etc. A wet cake comprising the crystalline polymorph may be washed with cold ethyl acetate prior to drying. 1.2 volumes of cold ethyl acetate may be used.

The process may further comprises the step of

(v) slurrying the third mixture prior to isolating the crystalline polymorph in step (iv). This may increase the yield of Form 1 from the process.

The slurrying in step (v) may be conducted at an ambient temperature, such as a temperature of about 25 °C or less, and preferable above 0 °C. That slurrying step may be conducted for a time period of at least about 1 hour, such as at least 2 hours, for instance at least 4 hours, at least 8 hours, at least 12 hours, at least one 1 , at least 2 days, at least 1 week at least 2 weeks.

A specific process for forming Form 1 comprises the steps of

(i) mixing the compound of Formula (I) with a suitable solvent to form the first mixture (which may be a solution);

(ii) heating the first mixture to a temperature of at least about 58 °C to form the second mixture, cooling the second mixture to a temperature in a range of from about 53°C to about 58°C preferably over a period of at least 20 minutes, and preferably holding the second mixture at the temperature in a range of from about 53 °C to about 58 °C for at least 30 minutes;

(iii) cooling the mixture to a temperature in a range of from about 0 °C to about 5 °C preferably over a period of at least 4 hours; and

(iv) isolating the crystalline polymorph, preferably by filtration, e.g. centrifuge filtration, at a temperature in a range of from about 0 °C to about 5 °C.

Form 1 may be prepared as described in the examples, for instance in Example 3-A, Example 4, or Example 12.

Form 2 may demonstrate no degradation (for example, less than 0.01 %, less than 0.1 %, less than 0.5% by weight) for at least 3 months under accelerated conditions (for example, 40°C and 75% RH), for at least 6 months under accelerated conditions, or for at least 9 months under accelerated conditions.

Form 2 may demonstrate no degradation for at least 3 months under ambient conditions (closed vial at approximately 25 °C), for at least 4 months under ambient conditions, for at least 5 months under ambient conditions, for at least 6 months under ambient conditions, for at least 9 months under ambient conditions, for at least 12 months under ambient conditions, for at least 24 months under ambient conditions, for at least 36 months under ambient conditions, or for at least 48 months under ambient conditions.

Form 2 may exhibit superior processability (including, for example, flow, blending, and compressibility) compared to amorphous 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide.

Without wishing to be bound by theory, such increased processability is believed to be due to its morphology. Additionally, consistency in compressibility of Form 2 may be superior to that of amorphous 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 , 4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide due to the variability of hardness of the amorphous material resulting from exposure to variable ambient humidity conditions.

Form 2 of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide exhibits decreased hygroscopicity compared to other solid state forms as determined by dynamic vapor sorption (DVS) studies. Form 2 may uptake less than 1 % water weight at 90% RH, less than 0.5% water weight at 90% RH, less than 0.3% water weight at 90% RH, less than 0.2% water weight at 90% RH, or less than 0.1 % water weight at 90% RH. This property of low hygroscopicity can aid in the preparation of solid pharmaceutical dosage forms.

Crystalline polymorph of Form 2 may be isolated with higher purity compared to other solid state forms, in particular the amorphous form.

The crystalline polymorph comprising Form 2 may have a particle size of 5-50 microns. The average particle size may be at least 10 microns, at least 15 microns, at least 20 microns, at least 25 microns, at least 30 microns, at least 35 microns, at least 40 microns, at least 50 microns, or at least 60 microns. It may have an average particle size of up to 35 microns, up to 40 microns, up to 45 microns, up to 50 microns, up to 60 microns, up to 70 microns, or up to 100 microns. The crystalline polymorph having an average diameter of greater than 5 microns or greater than 10 microns may have improved stability parameters compared to smaller diameters.

For any method disclosed herein that includes discrete steps, the steps may be conducted in any feasible order. And, as appropriate, any combination of two or more steps may be conducted simultaneously.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. All numerical values, however, inherently contain a range necessarily resulting from the standard deviation found in their respective testing measurements.

The invention will now be further illustrated by the following non-limiting examples. The specific examples below are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present invention to its fullest extent. All references and publications cited herein are hereby incorporated by reference in their entirety.

EXAMPLES

Experimental Methods

X-Ray Powder Diffraction (XRPD)

XRPD analyses were conducted using a D2 PHASER Cabinet X-Ray diffractometer (Bruker Corporation, Billerica, MA) operating with a Cu-Ka radiation source (1.54056 A), theta-two theta (Q/2Q) goniometer, 4 degree (°) soller, 0.6 millimeter (mm) divergent slits, and using a LYNXEYE detector (Bruker Corporation, Billerica, MA).

Samples were run under ambient conditions on a flat, low background specimen holder. Approximately 5 milligrams (mg)of material was placed on the holder and flattened with a microscope slide. The sample was rotated on its own plane during analysis. The scanning parameters were as follows: coupled Q/2Q in a range from 4° to 40 °2Q (±0.05 °2Q); 0.05 °2Q step size; 0.1 second dwell time; 30 rotations per minute (rpm) variable rotation. Peak assignment analyses were performed using Bruker DIFFRAC. SUITE EVA software.

The skilled person is aware that an X-ray powder diffraction pattern may be obtained that has one or more measurement errors depending on measurement conditions (such as equipment, sample preparation or instrument used). In particular, it is generally known that intensities in an X-ray powder diffraction pattern may fluctuate depending on measurement conditions and sample preparation. For example, the skilled person will realize that the relative intensity of peaks can be affected by, for example, grains above 30 microns in size and non-unitary aspect ratios, which may affect analysis of samples. The skilled person will also realize that the position of reflections can be affected by the precise height at which the sample sits in the diffractometer and the zero calibration of the diffractometer. The surface planarity of the sample may also have a small effect. Hence a person skilled in the art will appreciate that the diffraction pattern data presented herein is not to be construed as absolute (for further information see Jenkins, R & Snyder, R. L.‘Introduction to X-Ray Powder Diffractometry,’ John Wiley & Sons, 1996). Therefore, it shall be understood that the crystalline forms of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide are not limited to the crystals that provide X-ray powder diffraction patterns identical to the X-ray powder diffraction patterns described herein and any crystals providing X-ray powder diffraction patterns substantially the same as the X-ray powder diffraction patterns described herein fall within the scope of the present invention.

Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC) TGA/DSC analysis was conducted on a TGA/DSC 1 (Mettler Toledo, Columbus, OH) equipped with a 34 position autosampler. Samples typically contained between 2 mg and 6 mg; the sample was loaded onto a pre-tared aluminum crucible crimped with an aluminum lid and subsequently punctured on the instrument prior to sample analysis. Samples were heated at 10 degrees Celsius per minute (°C/min) from 35 °C to 300 °C. A purge of dry nitrogen was maintained over the sample. A second, empty aluminum pan used as a reference. The instrument control and data analysis software was STARe thermal analysis software (version 12.10).

The skilled person is aware that a DSC thermogram may be obtained which has one or more measurement errors depending on measurement conditions (such as equipment, sample preparation or instrument used). In particular, it is generally known that onset and/or peak temperatures may fluctuate depending on measurement conditions and sample preparation. Accordingly, it will be understood that the onset and/or peak temperature values of the DSC may vary slightly from one instrument to another, one method to another, from one sample preparation to another, and depending on the purity of the sample, and so the values quoted are not to be construed as absolute. Therefore, it shall be understood that the crystalline forms of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide are not limited to the crystals that provide DSC thermograms identical to the thermograms herein and any crystals providing thermograms substantially the same as the thermograms described herein fall within the scope of the present invention. As used herein, "substantially the same" when referring to a DSC thermogram means that a crystalline form provides a melt onset that is within ± 1 °C of the value shown in the thermograms referenced herein.

Fourier Transform Infrared Spectroscopy (FT-IR) Analysis

FT-IR Analysis was conducted using a Tensor 37 FT-IR spectrometer (Bruker Corporation, Billerica, MA) with MidIR source and DTGS detector and equipped with a ZnSe single bounce attenuated total reflectance (ATR) accessory. The sample for FT-IR analysis was prepared by placing samples onto crystal by completely filling sample trough. A pressure clamp was hand tightened to press powder into crystal. Data were collected from 4000 cm ¹ to 600 cm ¹.

The skilled person is aware that a FT-IR spectrum may be obtained that has one or more measurement errors depending on measurement conditions (such as equipment, sample preparation or instrument used). Therefore, it shall be understood that the crystalline forms of 6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide are not limited to the crystals that provide FT-IR spectra identical to the spectra herein and any crystals providing FT-IR spectra substantially the same as those described herein fall within the scope of the present invention.

Photomicroscopy Analysis

Photomicroscopy was performed using a VHX-600K (Keyence Corporation, Itasca, IL) digital microscope. A small amount of each sample was placed on a glass slide, mounted in mineral oil and covered with a glass slip. The individual particles were dispersed as well as possible. The sample was viewed with appropriate magnification (100-1000x).

Particle Size Distribution

Particle size analysis was performed using a HELOS laser diffraction unit with an ASPIROS dry feeder (Sympatec Gmbh, Clausthal-Zellerfeld, Germany). Analysis was performed at 2.0 bar. Results were collected using R4 or R6 detectors. Data collection trigger was optical concentration of greater than 2%. The Fraunhofer model was used for calculation.

All analysis were performed at room temperature (approximately 22°C) unless otherwise stated.

Dynamic Vapor Sorption (DVS)

Sorption isotherms were obtained using a DVS Advantage System, controlled by DVS Advantage Control software version 2.1.3.1. The sample temperature was maintained at 25 °C by instrument controls. The humidity was controlled by mixing streams of dry and wet nitrogen, with a total flow rate of 200 milliliters (ml_) per minute. The relative humidity (RH) was measured near the sample (dynamic ranges of 0% to 98% RH). The weight change of the sample as a function of % RH was constantly monitored by the microbalance (sensitivity 0.0001 mg).

About 10 mg of sample was placed in a tared clear bottomed pan. The sample was loaded and unloaded at 0% RH and 25°C. A moisture sorption isotherm was performed as outlined in Table 1. Data analysis was performed in Microsoft Excel using DVS Analysis Suite version 6.2.0.9. Images were taken after each % RH interval. Table 1 - Method Parameters for DVS Experiments

Table 2 - Solvent abbreviations

Example 1

Preparation of 6-{4-[1-(propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N-(pyridin- 4-yl)pyridine-2-carboxamide

A synthetic pathway for the preparation of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]- 1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide is provided in FIG. 1. Different isolation conditions can be used in order to obtain Form 1 of 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide.

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide was prepared through a series of organic synthesis reactions starting from 6-chloropicolinic acid, 4-aminopyridine, homopiperizine, 1 - isopropyl-4-piperidone and commercially available chemicals.

Step 1 - Preparation of Stage 1

To a vessel was added dichloromethane (10 volumes), 6-chloropicolinic acid (1.0 equivalent (eq)), and N,N-dimethylformamide (0.1 volumes). Oxalyl chloride (1.1 eq) was added, followed by a dichloromethane line rinse, while maintaining the temperature at 15°C - 25 °C. The mixture was stirred for at least 1 hour and then sampled for completion. The batch was cooled to 0°C - 5°C and charged with triethylamine (2.5 eq) and 4-aminopyridine (1.1 eq). The batch temperature was adjusted to 15°C - 25°C, stirred for at least 3 hours then sampled for completion. Dichloromethane (5 volumes) and water (3.3 volumes) are added, the mixture was stirred for at least 30 minutes and allowed to settle. The organic layer was discharged into drums and the aqueous layer was re-extracted with dichloromethane (5 volumes). The combined organic layers are washed with water (3.3 volumes) followed by 5% sodium hydrogen carbonate solution (5 volumes). This solution was transferred to another vessel and used in Stage 2 of the sequence.

Step 2 - Preparation of Stage 2 The solution prepared by Step 1 was concentrated down to minimum with stirring by distilling dichloromethane at atmospheric pressure. N,N- dimethylacetamide (5.3 volumes) was added and the mixture was concentrated under reduced pressure at not more than 45°C until all the dichloromethane has been removed. The batch temperature was adjusted to 40°C - 45°C and then homopiperazine (3.3 eq) was added followed by a N,N-dimethylacetamide line rinse (1.5 volumes). The batch was heated to 1 10°C - 120°C and stirred for a minimum of 3 hours. The batch was cooled to not more than 70°C before the addition of isopropanol (also known as Isopropyl Alcohol or IPA) (5 volumes) and then sampled for completion. Additional IPA was charged into the batch (6 volumes), the mixture was warmed back to 70°C - 75°C for at least 1 hour, then cooled back to 0°C - 5°C, and then stirred for a minimum of 2 hours. The batch was then transferred to the filter-dryer and washed with cold IPA (1.6 volumes). The batch was slurried on the filter in IPA (6.7 volumes) at 45°C - 55°C. The batch was washed with additional IPA (1.6 volumes) and dried for at least 12 hours at not more than 45°C until the Loss On Drying (LOD) was not more than 2%.

LOD was determined by drying 0.9 gram (g) - 1.1 g of sample, accurately weighed, in a vacuum oven at 45°C to constant weight.

Step 3 - Preparation of Stage 3

To a vessel was added dichloromethane (10 volumes) and the solution from Step 2 (0.5 eq.). A solution of 0.5 M sodium hydroxide was added (5 volumes) and the mixture was stirred at 20°C - 30°C for at least 30 minutes. The bottom organic layer was discharged to drums and the aqueous layer was re-extracted with more dichloromethane (2.5 volumes). The bottom organic layer was discharged to drums and the aqueous layer to waste. The above operation was repeated on the second 0.5 equivalent of the solution from Step 2. The combined organic layers are added back to the vessel and dichloromethane was distilled at atmospheric pressure to concentrate the batch down to approximately 1 1 volumes. The batch temperature was adjusted to 15°C - 25°C and then 1 - isopropyl-4-piperidone (1.25 eq.) and acetic acid (1.2 eq.) were added, followed by a dichloromethane line rinse. The mixture was stirred for at least 30 minutes and then charged with sodium triacetoxyborohydride (1.5 eq.) and additional dichloromethane (1.8 volumes). The mixture was stirred for not less than 5 hours before sampling for reaction completion. The reaction was quenched by charging 1 M sodium carbonate solution (8.3 volumes) over 1 hour and stirring for an additional 2 hours. The layers are separated and the organic phase was washed with water (3 volumes). The organic phase was transferred to a separate vessel and dichloromethane was distilled at atmospheric pressure down to approximately 3.6 volumes. The vessel was charged with methyl ethyl ketone (11 volumes) and distilled at atmospheric pressure down to a level of 5 volumes. The mixture was cooled to 0°C - 5°C, stirred for 1 hour and then transferred to the filter. The wet cake was washed with methyl ethyl ketone (1.1 volumes) at 0°C - 5°C and dried for at least 12 hours at room temperature until the LOD (method discussed above) was not more than 2%.

Step 4 - Final Product

The amorphous form of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide can be obtained by heating 6-{4-[1 -(Propan- 2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide to a molten state, and then cooling rapidly to solidify.

Example 2

Methods of preparing crystalline 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 , 4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1 and Form 2 are as follows.

Room Temperature Slurry Conversion

A number of solvents were used to convert a slurry into Form 1 or Form 2 at room temperature (approximately 22 °C). For each solvent, a standard 4 ml_ crimp cap vial was loaded with 80 mg of amorphous 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide, 2000 pL of solvent, and a magnetic stir bar. The vials were then crimped closed and mixed. If material dissolved after 24 hours, the sample was moved to other precipitation techniques. If material did not dissolve, samples continued to slurry at room temperature for two weeks.

The presence of Form 1 or Form 2 was confirmed by XRPD. Where applicable, the solids were analysed both wet (that is, without prior drying) and dry.

Solvents in which Form 1 was form when analysed wet include DMSO, ethylene glycol, DMF, acetonitrile, IPA, MEK, MIBK, 1 ,2-dimethoxyethane, IPAc, 1 ,2- xylene, toluene, 1 ,4-dioxane, and heptane. Solvents in which Form 1 was form when analysed dry include DMF, acetonitrile, acetone, IPA, MEK, MIBK, 1 ,2- dimethoxyethane, methylTHF, methyl acetate, ethylacetate, IPAc, diethyl ether, MTBE, 1 ,2-xylene, toluene, 1 ,4-dioxane, and heptane.

Form 2 was formed when water was used as the solvent, when analysed wet or dry.

These results suggest that Form 1 is thermodynamically stable at room temperature

High Temperature Slurry Conversion

In addition to the slurry conversion at room temperature, slurry experiments were performed at 75°C. This temperature was selected because it is above the experimentally determined transition temperature. For each solvent, a standard 4 ml_ crimp cap vial was loaded with 200 mg of amorphous 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide, 200 pL to 1500 pl_ of solvent (depending on the visual solubility), and a magnetic stir bar. The vials were then crimped closed and slurried for two weeks.

At the end of the slurry, solids were dried and analyzed by XRPD. Solvents which produced Form 2 include DMSO, ethylene glycol, DMF, NMP, MIBK, 1 ,2- dimethoxyethane, IPAc, 2-ethoxyethanol, 1 ,2-xylene, toluene, 1 ,4-dioxane, and heptane. All solvents produced Form 2 suggesting that Form 2 is thermodynamically stable at 75 °C.

Room Temperature Evaporation

For each experiment, solution or supernatant from the room temperature slurry conversion was placed into a 4 ml_ vial. The amount of solution was based upon concentration values ensuring that enough solution was used to generate ~5 mg of material for analysis by XRPD. Vial was crimped and punctured with a needle and allowed to slowly evaporate at ambient temperature (approximately 22°C). until dryness. If evaporation was too slow to be complete at two weeks the vial was uncapped. Solids from the evaporation were harvested and analysed by XRPD.

Form 1 was formed using DMF, DCM, THF (in combination with Form 2), methylTHF, 2-ethoxyethanol, 1 ,2-xylene, toluene, and 1 ,4-dioxane.

Form 2 was formed using ethanol, THF (in combination with Form 1 ), and chloroform.

Elevated Temperature Evaporation

For each experiment, solution or supernatant from the room temperature slurry conversion was placed into a 4 ml_ vial. The amount of solution was based upon concentration values ensuring that enough solution was used to generate ~5 mg of material for analysis by XRPD. Vial was crimped and punctured with a needle and allowed to evaporate at 75°C until dryness. If evaporation was to slow to be complete at two weeks the vial was uncapped. Samples were moved to room temperature once dry to prevent degradation when applicable. Solids from the evaporation were harvested and analysed by XRPD

Form 1 was formed using ethanol, MEK, 1 ,2-dimethoxyethane, methylTHF (in combination with Form 2), methyl acetate, and ethyl acetate.

Form 2 was formed using THF, methylTHF (in combination with Form 1 ), chloroform, 1 ,2-xylene, and 1 ,4-dioxane. 5 °C to Evaporation

Form 1 was formed using acetonitrile, acetone, MEK, MIBK, 1 ,2- dimethoxyethane, ethyl acetate, diethyl ether, and MTBE.

Form 2 was formed using methyl acetate.

5°C Precipitation

A solution or supernatant comprising 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 , 4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide was added to a 4 ml_ vial. Each vial was capped and placed in a 5 °C chamber for one week. Solids formed were harvested wet and analysed by XRPD. If there were not enough solids for analysis after one week or the solvent did not evaporate, then the vial was moved to ambient temperature (approximately 22°C) evaporation: the vial was capped with tinfoil, punctured with a needle, and allowed to slowly evaporate at room temperature until dryness. Solids from the evaporation were harvested and analysed by XRPD. Where applicable, the solids were analyzed both wet (that is, without prior drying) and dry.

Solvents in which Form 1 was form when analysed wet include DMF, and 1 ,2- xylene. Solvents in which Form 1 was form when analysed dry include IPAc, 1 ,2-xylene, and toluene. These samples were consistently Form 1.

Crash Crystallization with Anti-Solvent Addition

For each experiment, a solution or supernatant comprising 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide was placed in a 4 ml_ vial. Anti-solvents were selected based on miscibility with the solvent and solubility of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide. Appropriate anti-solvent was added dropwise to the solution/supernatant until approaching precipitation. Samples were then kept at a temperature of 2 °C to 8 °C overnight. Solids were harvested and analysed by XRPD. Where applicable, the solids were analysed both wet (that is, without prior drying) and dry. Solvents in which Form 1 was form when analysed wet include methanol (with water as the anti-solvent) and THF, and 1 ,4-dioxane (with heptane as the anti solvent).

Solvents in which Form 1 was form when analysed dry include methanol (with water as the anti-solvent) and THF, methylTHF, methyl acetate, 1 ,2-xylene, toluene, and 1 ,4-dioxane (with heptane as the anti-solvent).

Solvents in which Form 2 was form when analysed wet include ethylene glycol, and DMF (with water as the anti-solvent).

Anti-Solvent Addition and Evaporation

Solvents in which Form 1 was form when analysed wet include ethanol (in combination with Form 2), and I PA (with heptane as the anti-solvent).

Solvents in which Form 1 was form when analysed dry include ethanol (in combination with Form 2), IPA, MEK, DCM, 1 ,2-domethoxyethane (in combination with Form 2), ethyl acetate, IPAc, 2-ethoxyethanol, and chloroform (with heptane as the anti-solvent).

Solvents in which Form 2 was form when analysed wet include DMSO (with water as the anti-solvent), and ethanol (in combination with Form 1 , and with heptane as the anti-solvent).

Solvents in which Form 2 was form when analysed dry include ethanol (in combination with Form 1 ), and 1 ,2-domethoxyethane (in combination with Form 1), both of which with with heptane as the anti-solvent.

Example 3-A

Preparation of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N-(pyridin- 4-yl)pyridine-2-carboxamide Form 1 6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide of Form 1 was prepared as described in Example 1 except that Step 4 was performed as follows instead of as described in Example 1.

Step 4 - Final Product

A vessel was charged with purified water (1.5% of ethyl acetate charge), ethyl acetate (16 volumes), and the solution from Step 3 (1.0 eq). The mixture was warmed to 50 °C to 55 °C and stirred for at least 1 hour. The resulting solution was transferred to the crystallisation vessel via an in-line filter followed by a warm ethyl acetate line rinse. The batch was cooled to not more than 25 °C and then distilled under reduced pressure down to 5 volumes. This mixture was stirred at 15 °C to 25 °C for not less than 12 hours, cooled to 0 °C to 5 °C, stirred for at least 1 hour before transferring to the filter. The wet cake was washed with additional cold ethyl acetate (1.2 volumes) and dried on the filter for not less than 6 hours until drying specification was met.

The 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide of Form 1 produced was >98% pure with an overall yield for the entire process of 54%.

Example 3-B

Preparation of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N-(pyridin- 4-yl)pyridine-2-carboxamide Form 2

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2 was prepared as described in Example 1 except that Step 4 was performed as follows instead of as described in Example 1.

Step 4 - Final Product

The 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide resulting from Step 3 was dissolved methyl ethyl ketone and then the solvents were removed in vacuo. The residue was dried in a vacuum oven overnight. The 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide Form 2 produced was 98.6% pure.

Example 4

Conversion of 6-{4-[1 -(propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide of Form 1 to 6-{4-[1-(propan-2-yl)piperidin-4-yl]-1,4- diazepan-1-yl}-N-(pyridin-4-yl)pyridine-2-carboxamide of Form 2 and vice versa

The transition temperature for two enantiotropic polymorphs is the temperature at which solubilities of the two forms are equal. This property enable estimation of the transition temperature based on thermodynamic properties using a simplified solubility equation (Yalkowski, Solubility and Solubilization in Aqueous Media, 1999, pages 62-64) and assuming the influence of heat capacity difference between solid and liquid is negligible between forms:

Rearranging yields the following equation for estimate of transition temperature:

Where T₀ is the melting point in Kelvin, DH is the heat of fusion in kilojoules per mole, and the subscripts 1 and 2 refer to the Forms of the two polymorphs.

The transition temperature was calculated from the melting points and heats of fusion for each form, measured by DSC, resulting in a calculated transition temperature of 69°C.

Competitive slurry experiments were conducted in four different solvents at three temperature conditions to determine if experimental results support the calculated transition temperature of the enantiotropic pair. For each solvent, Form 1 and Form 2 were weighed separately into 4 ml_ vials. The indicated solvent was added as shown in Tables 3, 4, and 5. Table 3 - 60°C Competitive Slurry

Table 4 - 70°C Competitive Slurry

Table 5 - 80°C Competitive Slurry

A magnetic stir bar was added to each vial, and each vial was fitted with a screw cap. Vials were placed in a REACTI-THERM (ThermoFisher Scientific, Minneapolis, MN), heated to 60 °C, 70 °C, or 80 °C, and mixed. After the vials were slurried for 19.25 hours, 22.75 hours, or 17.5 hours, respectively, the samples were centrifuge filtered. Solids from the slurry were harvested and analysed by XRPD.

At 60 °C, conversion from Form 2 to Form 1 was apparent in two of the four solvents. In addition, when Form 1 was used as the input material, Form 1 returned, suggesting Form 1 is thermodynamically stable at 60°C. At 70 °C, primarily the form of the input material returned at the end of the slurry experiment. However, slight conversion from Form 1 to Form 2 was apparent in water. These results suggest the transition temperature of the enantiotropic pair is near 70°C. A Form 1 to Form 2 conversion observed in water suggests that the transition temperature maybe slightly lower than 70°C.

At 80 °C, conversion from Form 1 to Form 2 was apparent in two of the four solvents. In addition, when Form 2 was used as the input material, primarily Form 2 returned, suggesting Form 2 is thermodynamically stable at this temperature.

The isolation of Form 1 at temperatures below 70 °C and the isolation of Form 2 at temperatures above 70°C is consistent with the calculated transition temperature of 69 °C.

Example 5-A

XRPD Analysis of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N- (pyridin-4-yl)pyridine-2-carboxamide of Form 1

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 1 , made as described in Example 3-A was subjected to XRPD analysis.

Exemplary results of the XRPD scan are shown in FIGS. 2 and 4, and the peak assignments are as noted above.

Example 5-B

XRPD Analysis of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N- (pyridin-4-yl)pyridine-2-carboxamide of Form 2

6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide Form 2 made as described in Example 3-B, was subjected to XRPD analysis as described above. Exemplary results of the XRPD scan are shown in FIGS. 3 and 4, and the peak assignments are as noted above.

Example 6

TGA and DSC analysis of 6-{4-[1 -(propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}- N-(pyridin-4-yl)pyridine-2-carboxamide of Forms 1 and 2

Thermal properties of crystalline 6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4- diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base made as described in Examples 3-A and 3-B, were assessed by TGA and DSC analysis.

FIG. 5 shows exemplary results of DSC analysis of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base Form 1. FIG. 6 shows exemplary results of DSC analysis of crystalline 6- {4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide free base Form 2. Samples were heated at a rate of 10 °C per minute from 35 °C to 300 °C.

As can be seen in FIG. 5, Form 1 demonstrates a distinct melt endotherm at approximately 141 °C to 142 °C, followed by recrystallization and melting of a second polymorph (Form 2) at 157 °C to 159 °C. As can be seen in FIGS. 5-6, Form 2 melts at approximately 157 °C to 159 °C. Weight loss was minimal across the melt events. Recrystallization into Form 2, as visible in the thermograms following the melt of Form 1 , indicates an endothermic event.

An exemplary combined TGA/DSC profile of Form 1 and Form 2 is provided in FIG. 7.

Example 7

FT-IR Analysis of crystalline 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1- yl}-N-(pyridin-4-yl)pyridine-2-carboxamide FT-IR analysis was performed with several batches of crystalline 6-{4-[1 - (propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2- carboxamide made as described in Examples 3-A and 3B.

As depicted in FIGS. 8-10, changes in frequencies and relative intensity of the band contours show clear differences between Form 1 and Form 2. The differences are most apparent in the 3300-3400 cm ¹ range which most likely corresponds to the amide N-H stretch. Differences were also observed in the 1560-1750 cm ¹ range which most likely corresponds to the amide C=0 carbonyl stretch.

Example 8

Photomicroscopy

Pictomicrographs of samples of crystalline 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]- 1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base (made as described in Examples 3-A and 3-B) were generated, providing insight into crystal morphology.

FIG. 1 1 shows a representative image of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base Form 1. Exemplary crystal shapes of Form 1 are plates.

FIG. 12 shows a representative image of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base Form 2. Exemplary crystal shapes of Form 2 are lathes.

Example 9

Particle Size Distribution Analysis

Laser diffraction data was collected for samples of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base (made as described in Examples 3-A and 3-B). Particle size data show reasonable correlation to the sizes of the particles observed during microscopy (for example, a d₅o (that is, the diameter at which 50% of a sample's mass is comprised of smaller particles, also referred to as a mass median diameter) of approximately 50 micrometers (pm) and a d₉₀ (that is, the diameter at which 90% of a sample's mass is comprised of smaller particles) of approximately 184 pm). This morphology and size range is expected to be conductive to ready flowability of powder, making the material suitable for drug production manufacturing.

Example 10

Moisture Uptake

DVS Data were collected for samples of crystalline 6-{4-[1 -(propan-2 - yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4-yl)pyridine-2-carboxamide free base (made as described in Examples 3-A and 3-B).

Batches typically displayed similar moisture sorption properties with moisture sorption uptakes of 0.0% to 0.5% at 90% RH. An exemplary sorption/desorption profile is shown in FIG. 13.

Example 11

Single Crystal Data for 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan-1-yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1

Crystals of 6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide Form 1 were made as described in Example 3-A. A colourless crystal of 6-{4-[1 -(Propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N- (pyridin-4-yl)pyridine-2-carboxamide Form 1 with dimensions of 0.200 x 0.100 x 0.050 mm was selected for analysis.

Data were collected on an Oxford Diffraction Supernova Dual Source, Cu at Zero, Atlas CCD diffractometer equipped with an Oxford Cryosystems Cobra cooling device. The data was collected using CuKa radiation. Structures were typically solved using either the SHELXS of SHELXD programs and refined with SHELXL program as part of the Bruker AXS SHELXTL suite (V6.10). Unless otherwise stated, hydrogen atoms attached to carbon were placed geometrically and allowed to refine with a riding isotropic displacement parameter. Hydrogen atoms attached to a heteroatom were located in a difference Fourier synthesis and were allowed to refine freely with an isotropic displacement parameter. Data collection and refinement parameters are shown in Table 6.

Table 6 - Data collection and refinement parameters

A conformational picture of 6-{4-[1 -(propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 - yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1 based on single crystal X-ray analysis is shown in FIG 14. 6-{4-[1 -(Propan-2-yl)piperidin-4-yl]-1 ,4-diazepan-1 - yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1 is orthorhombic, Pbca space group, with the following unit cell dimensions: a = 12.4413(5) A, b = 8.9899(5) A, c = 39.343(2) A, a = 90°, b = 90°, g = 90°. Additional crystal data is shown in Table 7.

Table 7 - Additional crystal data

Example 12

A method for crystallization of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan- 1-yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 1:

Dissolve 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide at 60°C-70°C in 1.5 % v/v water in ethyl acetate (12 volumes), transfer through an in-line filter to the crystallization vessel at not less than (NLT) 60°C and then cool to 53°C - 58°C (target 55°C - 56°C) over NLT 30 minutes and hold at 53°C - 58°C (target 55°C - 56°C) for NLT 1 hour. Cool to 0°C - 5°C over NLT 5-6 hours and hold at 0°C - 5°C for NLT 1 hour. Filter and rinse with 1 volume of 1.5 % v/v water in ethyl acetate at 0°C-5°C. Resulting product was analysed by FTIR and XRPD. Results are shown in Table 8.

Table 8 - Comparison of slow cooling stage 4 crystallizations

Example 13

A method for crystallization of 6-{4-[1-(Propan-2-yl)piperidin-4-yl]-1,4-diazepan- 1-yl}-N-(pyridin-4-yl)pyridine-2-carboxamide Form 2

Samples of 6-{4-[1 -(propan-2 -yl)piperidin-4-yl]-1 ,4-diazepan-1 -yl}-N-(pyridin-4- yl)pyridine-2-carboxamide were dissolved in 1.5 % v/v water in ethyl acetate, MEK or IPA (10 volumes) at 60°C and then rapidly cooled to 0°C-5°C in an ice bath (over less than (LT) 15 minutes), stirred at 0-5 °C for not less than (NLT) 1 hour, isolated by vacuum filtration and displacement washed with 1 volume of solvent at 0°C-5°C. The products (static pale yellow powders) were dried in a vacuum oven. Resulting products were analyzed by FTIR and XRPD. Results are shown in Table 9. Table 9 - Comparison of rapid cooling stage 4 crystallizations

The present invention is not intended to describe each disclosed embodiment or every implementation of the present invention. The description that follows more particularly exemplifies illustrative embodiments. In several places throughout the application, guidance is provided through lists of examples, which examples can be used in various combinations. In each instance, the recited list serves only as a representative group and should not be interpreted as an exclusive list.

Claims

Claims

1. A crystalline polymorph comprising the compound of Formula (I)

2. The crystalline polymorph of claim 1 , wherein the compound is a free base.

3. The crystalline polymorph of claim 1 or claim 2, wherein the polymorph consists of the compound of Formula (I).

4. The crystalline polymorph of any preceding claim, wherein the crystalline polymorph is crystalline Form 2.

5. The crystalline polymorph of any preceding claim, wherein the crystalline polymorph exhibits an XRPD pattern with peaks at 8.9°2Q ± 0.3°2Q, 15.9°2Q ± 0.3°2Q, 18.6°2Q ± 0.3°2Q, and 21.8°2Q ± 0.3°2Q..

6. The crystalline polymorph of claim 5, wherein the crystalline polymorph exhibits an XRPD pattern with peaks at 9.9°2Q ± 0.3°2Q.

7. The crystalline polymorph of any preceding claim, wherein the crystalline polymorph exhibits an XRPD pattern and wherein the XRPD pattern is substantially the same as the XRPD pattern of FIG. 3.

8. The crystalline polymorph of any preceding claim having a differential scanning calorimetry (DSC) thermogram exhibiting an endothermic event having a melt maxima temperature in the range of from about 157 °C to 159 °C.

9. The crystalline polymorph of any preceding claim having a differential scanning calorimetry (DSC) thermogram substantially the same as the DSC thermogram of FIG. 6.

10. A pharmaceutical composition comprising the crystalline polymorph of any preceding claim, and a pharmaceutically acceptable carrier and/or excipient.

1 1. The pharmaceutical composition of claim 10 comprising Form 1 of the compound of Formula (I).

12. The crystalline polymorph of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or claim 1 1 , for use in the treatment of a disease or condition responsive to reduction of CXCR4-mediated activity.

13. A method for treating a subject having a disease or condition responsive to reduction of CXCR4-mediated activity, the method comprising administering an effective amount of the crystalline polymorph of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or claim 1 1 , to a subject in need thereof.

14. The crystalline polymorph or pharmaceutical composition for use according to claim 12, or the method according to claim 13, wherein the disease or condition responsive to reduction of CXCR4-mediated activity is selected from the group consisting of cancer, HIV/AIDS, neuropathy, HIV related neuropathy, pain, inflammation, brain inflammation, neurodegeration, cognitive degeneration, diabetic retinopathy, age related macular degeneration, retinal neo- vascularisation, and viral infections.

15. The crystalline polymorph or pharmaceutical composition for use, or the method, according to claim 14, wherein the disease or condition is cancer which may be selected from the group consisting of cancer of the breast, lung (including non-small cell and small cell), pancreas, cervix, thyroid, kidney, bladder, ovary, prostate, skin (including melanoma), cancer of the Gl tract (including oesophageal, hepatic, colorectal and gastric cancers), oral squamous carcinoma, cancers of the blood including leukaemias such as B-CLL, AML, CML, ALL, lymphomas such as intraocular, non-Hodgkins and Hodgkins lymphomas, and multiple myeloma; neuroblastoma, cancers of the nervous system including cancer of the brain, glioma, glioblastoma, other astrocytomas, oligodendroglial tumour, meningioma, ependymoma, oligodendroglioma, medulloblastoma, and metastases into the CNS from peripheral cancers, preferably the cancer is a CNS cancer, for instance cancer of the brain, preferably the CNS cancer is glioma, more preferably the CNS cancer is selected from the group consisting of glioblastoma, other astrocytomas, oligodendroglial tumour, meningioma, ependymoma, oligodendroglioma, medulloblastoma, and metastases into the CNS from peripheral cancers, most preferably the CNS cancer is glioblastoma and astrocytoma, especially glioblastoma.

16. The crystalline polymorph or pharmaceutical composition for use, or the method, according to claim 14 or claim 15, wherein compound of Formula (I) is administered in combination with one or more CHK1 inhibitors and/or one or more anti-angiogenic agents, preferably with one or more CHK1 inhibitors.

17. The crystalline polymorph or pharmaceutical composition for use according to claim 12, or the method according to claim 13, wherein the use or method comprises an apheresis procedure for promoting release and mobilisation of stem cells, including haematopoietic and non-haematopoietic stem cells and progenitor stem cells prior to harvesting, optionally wherein (i) GCSF is used in the apheresis procedure; and/or (ii) the apheresis procedure is implemented prior to treatment of a subject by chemotherapy or radiotherapy, for reducing chemotherapy- or radiotherapy-induced leukopenia.

18. A process for preparing a crystalline polymorph according to any one of claims 1 to 12, the process comprising the steps of

(i’) mixing the compound of Formula (I) with a suitable solvent to form a first mixture;

(ii’) heating the first mixture to form a second mixture;

(iii’) cooling the second mixture to form a third mixture; and (iv’) isolating the crystalline polymorph.

19. The process of claim 18, wherein the step of heating the first mixture to form a second mixture comprises heating the first mixture to a temperature of greater than about 69 °C, such as from about 75 °C to about 150 °C, preferably to a temperature of from about 80 °C to about 120 °C, more preferably from about 85 °C to about 110 °C, even more preferably from about 90 °C to about 100 °C, and optionally holding the second mixture at that temperature for at least 30 minutes before cooling the second mixture in step (iii).

20. The process of claim 18 or claim 19, wherein the second mixture is a slurry.

21. The process of claim 20, wherein the slurry slurried for at least 1 hour, such as at least 3 hours, for example at least 6 hours, preferably at least 12 hours, more preferably at least 18 hours, even more preferably at least 24 hours, such as at least 36 hours, for instance at least 48 hours, e.g. at least 72 hours, or even at least 120 hours, for instance up to 168 hours.

22. The process of any one of claims 18 to 21 , wherein in step (iii’) solvent is cooled to a temperature in a range of 0 °C to 5 °C, for instance over a period of up to 20 minutes, and optionally being mixed at that temperature for at least 30 minutes.

23. The process of any one of claims 18 to 22, wherein in step (iv’) the crystalline polymorph is isolated by centrifuge filtration, for instance centrifuge vacuum filtration and displacement, or by vacuum filtration.

24. The process of any one of claims 18 to 23, wherein the suitable solvent is a polar solvent.

25. The process of any one of claims 18 to 23, wherein the suitable solvent comprises a solvent selected from the group consisting of water, DMSO, ethylene glycol, DMF, NMP, MIBK, 1 ,2-dimethoxyethane, IPAc, 2-ethoxyethanol, 1 ,2-xylene, toluene, 1 ,4-dioxane, and heptane.

26. The process of any one of claims 18 to 23, wherein the suitable solvent comprises at least one of ethyl acetate, metyl ethyl ketone, and isopropyl alcohol, preferably methyl ethyl ketone.

27. The process of any one of claims 18 to 26, wherein the suitable solvent comprises water.