Crystalline forms of potassium channel modulators

Abstract

Provided herein are crystalline and amorphous forms of a compound having structural formula (1). Also provided are pharmaceutical compositions comprising the crystalline and amorphous forms, methods for their manufacture, and uses thereof for treating a variety of diseases, disorders or conditions, associated with potassium channels.

Description

BACKGROUND

Among the ion channels, potassium channels are the most prevalent and diverse, being found in a variety of animal cells such as nervous, muscular, glandular, immune, reproductive, and epithelial tissue. These channels allow the flow of potassium in and/or out of the cell under certain conditions. These channels are regulated, e.g., by calcium sensitivity, voltage-gating, second messengers, extracellular ligands, and ATP-sensitivity.

Dysfunction of potassium channels and dysfunction from other causes which influence these potassium channels are known to generate loss of cellular control, altered physiological function, and disease conditions. Because of their ability to modulate ion channel function and/or regain ion channel activity, potassium channel modulators are being used in the pharmacological treatment of a wide range of pathological diseases and have the potential to address an even wider variety of therapeutic indications.

The small conductance calcium-activated potassium channels (SK channel) are a subfamily of Ca²⁺ activated K⁺ channels and the SK channel family contains 4 members—SK1, SK2, SK3, and SK4 (often referred to as intermediate conductance). The physiological roles of the SK channels have been especially studied in the nervous system, where for example they are key regulators of neuronal excitability and of neurotransmitter release, and in smooth muscle, where they are crucial in modulating the tone of vascular, broncho-tracheal, urethral, uterine or gastro-intestinal musculature.

Compound 1 is a small molecule modulator of potassium ion channels showing great therapeutic potential for treating a variety of diseases characterized by dysfunction of potassium ion channels and dysfunction from other causes which influence these potassium channels. Compound 1 is exemplified in U.S. Pat. No. 9,975,886, the contents of which are incorporated herein by reference, and has the structure:

The development of alternative forms of Compound 1 represents an attractive area to further the treatment of diseases such as those responsive to the modulation of the small conductance calcium-activated potassium subtype 2(SK2) channel.

SUMMARY

Provided herein are amorphous and crystalline forms of Compound 1.

Also provided herein are pharmaceutical compositions comprising one or more of the disclosed amorphous or crystalline forms of Compound 1.

Further provided is the use of one or more of the disclosed amorphous or crystalline forms of Compound 1 in the treatment diseases and/or disorders associated with the activity of potassium channels.

Processes for making the disclosed amorphous and crystalline forms are also provided.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form B.

FIG. 2 depicts the combined thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram for crystalline Form B.

FIG. 3 depicts single x-ray diffraction (SCXRD) analyses of the crystal structure of Form B: A-E the Oak Ridge Thermal Ellipsoid Plot (ORTEP) structural illustrations of 3A the asymmetric unit of Form B single crystal, 3B the asymmetric unit of Form B single crystal consisting of two Compound 1 molecules, 3C the unit cell of Form B single crystal consisting of eight Compound 1 molecules, 3D hydrogen bonds in the Form B single crystal structure, and 3E the 3D packing of the unit cell viewed along c-axis in Form B single crystal; and 3F calculated and experimental XRPD patterns of Form B.

FIG. 4 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form C.

FIG. 5 depicts the combined thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram for crystalline Form C.

FIG. 6 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form D.

FIG. 7 depicts the combined thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram for crystalline Form D.

FIG. 8 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form E.

FIG. 9 depicts the combined thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram for crystalline Form E.

FIG. 10 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form F.

FIG. 11 depicts the combined thermogravimetric analysis (TGA) thermogram and differential scanning calorimetry (DSC) thermogram for crystalline Form F.

FIG. 12 depicts an x-ray powder diffraction (XRPD) pattern for crystalline Form A (a mixture of crystalline Form B and C).

FIG. 13 depicts the x-ray powder diffraction (XRPD) patterns for amorphous Form of Compound 1.

FIG. 14 depicts the differential scanning calorimetry (DSC) thermograms for amorphous Form of Compound 1.

DETAILED DESCRIPTION

1. Definitions

As used herein, “crystalline” refers to a solid form of a compound wherein there exists long-range atomic order in the positions of the atoms. The crystalline nature of a solid can be confirmed, for example, by examination of the X-ray powder diffraction pattern.

As used herein, a “single crystalline form” means that the recited compound, i.e., Compound 1, is present as a single crystal or a plurality of crystals in which each crystal has the same crystal form (e.g., Form B, C, D, E, or F). When the crystal form is defined as a specified percentage of one particular single crystalline form of the compound, the remainder is made up of amorphous form and/or crystalline forms other than the one or more particular forms that are specified. In one embodiment, e.g., a disclosed crystalline form is at least 60% a single crystalline form, at least 70% a single crystalline form, at least 75% a single crystalline form, at least 80% a single crystalline form, least 85% a single crystalline form, at least 90% a single crystalline form, at least 95% a single crystalline form, at least 96% a single crystalline form, at least 97% a single crystalline form, at least 98% a single crystalline form, or at least 99% a single crystalline form by weight. Percent by weight of a particular crystal form is determined by the weight of the particular crystal form divided by the sum weight of the particular crystal, plus the weight of the other crystal forms present plus the weight of amorphous form present multiplied by 100%.

Unless otherwise specified, Form B, Form C, Form D, and Form F are each single crystalline forms as defined herein. Therefore, “Form B”, “single crystalline Form B”, “single crystalline Form B”, or “single solid” when referring to “Form B” are used interchangeably. The same applies for Form C, Form D, and Form F.

Chemical purity refers to extent by which the disclosed form is free from materials having different chemical structures. Chemical purity of the compound in the disclosed crystal forms means the weight of the compound divided by the sum of the weight of the compound plus materials/impurities having different chemical structures multiplied by 100%, i.e., percent by weight. In one embodiment, the compound in one or more of the disclosed crystalline forms has a chemical purity of at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by weight.

The term “amorphous” refers to a solid that is present in a non-crystalline state or form. Amorphous solids are disordered arrangements of molecules and therefore possess no distinguishable crystal lattice or unit cell and consequently have no definable long range ordering. Solid state ordering of solids may be determined by standard techniques known in the art, e.g., by X-ray powder diffraction (XRPD) or differential scanning calorimetry (DSC). Amorphous solids can also be differentiated from crystalline solids e.g., by birefringence using polarized light microscopy.

The term “anhydrous” and “anhydrate” are used interchangeably and mean that the referenced crystalline form has substantially no water in the crystal lattice, e.g., less than 0.1% by weight as determined by Karl Fisher analysis.

The term “solvate” refers to the physical association of a crystalline or amorphous form of Compound 1 as described herein with one or more solvent molecules. In certain instances, the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid. Exemplary solvates include hydrates, isopropanolates, ethanolates, methanolates, dichloromethanolates, and the like.

Compound 1 refers to a compound having the following structural formula:

The 2-theta values of the X-ray powder diffraction patterns for the crystalline forms described herein may vary slightly from one instrument to another and also depending on variations in sample preparation and batch to batch variation due to factors such as temperature variation, sample displacement, and the presence or absence of an internal standard. Therefore, unless otherwise defined, the XRPD patterns/assignments recited herein are not to be construed as absolute and can vary ±0.2 degrees. It is well known in the art that this variability will account for the above factors without hindering the unequivocal identification of a crystal form. Unless otherwise specified, the 2-theta values provided herein were obtained using Cu Kα1 radiation.

Temperature values, e.g., for DSC peaks herein may vary slightly from one instrument to another and also depending on variations in sample preparation, batch to batch variation, and environmental factors. Therefore, unless otherwise defined, temperature values recited herein are not to be construed as absolute and can vary ±5 degrees or ±2 degrees.

“Substantially the same XRPD pattern” or “an X-ray powder diffraction pattern substantially similar to” a defined figure means that for comparison purposes, at least 90% of the peaks shown are present. It is to be further understood that for comparison purposes some variability in peak intensities from those shown are allowed, such as ±0.2 degrees.

As used herein the terms “subject” and “patient” may be used interchangeably, and means a mammal in need of treatment, e.g., companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats and the like) and laboratory animals (e.g., rats, mice, guinea pigs and the like). Typically, the subject is a human in need of treatment.

The term “pharmaceutically acceptable carrier” refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

The terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, reducing the likelihood of developing, or inhibiting the progress of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors), i.e., prophylactic treatment. Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

The term “effective amount” or “therapeutically effective amount” includes an amount of a compound described herein that will elicit a biological or medical response of a subject.

2. Exemplary Forms

Provided herein is a crystalline Form B of a compound having the structural formula:

In one aspect, crystalline Form B is characterized by at least three x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by at least five x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by at least six x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 18.9°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 18.9°, 20.1°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 9.2°, 10.9°, 13.1°, 14.6°, 15.5°, 17.7°, 18.9°, 20.1°, 20.6°, 21.4°, 22.0°, 22.3°, and 23.2°. In other aspects, crystalline Form B is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 9.2°, 10.9°, 11.5°, 12.2°, 13.1°, 13.7°, 14.0°, 14.6°, 15.5°, 16.9°, 17.7°, 18.4°, 18.9°, 19.1°, 20.1°, 20.6°, 20.9°, 21.4°, 22.0°, 22.3°, 23.2°, 24.0°, 24.5°, 25.2°, 25.9°, 26.4°, 27.1°, 28.1°, 28.9°, 29.7°, 33.9°, 34.7°, 35.2°, 37.3°, and 38.9°. In other aspects, crystalline Form B is characterized by at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen, or at least twenty x-ray powder diffraction peaks at 2Θ angles selected from those in Table 16. In other aspects, crystalline Form B is characterized by an XRPD (x-ray powder diffraction) substantially similar to FIG. 1. In other aspects, crystalline Form B is characterized by a Thermogravimetric analysis (TGA) of a 0.1% weight loss up to 150° C. or Differential Scanning calorimetry (DSC) of a sharp endotherm at 170.6° C. (onset temperature), or both, wherein the crystalline Form B may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form B is characterized by a Thermogravimetric analysis (TGA) or Differential Scanning calorimetry (DSC) substantially similar to FIG. 2, wherein the crystalline Form B may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form B crystallizes in the P2₁/c monoclinic space group, wherein the crystalline Form B may also comprise XRPD peaks at 2Θ angles selected from those above and/or the TGA or DSC values or figures recited above. In other aspects, crystalline Form B is anhydrous, wherein the crystalline Form B may also comprise XRPD peaks at 2Θ angles selected from those above, the TGA or DSC values or figures recited above, and/or crystallize in the P2₁/c monoclinic space group.

Also provided herein is a crystalline Form C of a compound having the structural formula:

In one aspect, crystalline Form C is characterized by at least three x-ray powder diffraction peaks at 2Θ angles selected from 4.2°, 8.5°, 17.0°, 20.2°, and 21.4°. In other aspects, crystalline Form C is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 4.2°, 8.5°, 17.0°, 20.2°, and 21.4°. In other aspects, crystalline Form C is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 4.2°, 8.5°, 17.0°, 20.2°, and 21.4°. In other aspects, crystalline Form C is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 4.2°, 4.5°, 8.5°, 9.1°, 9.7°, 10.4°, 13.1°, 13.7°, 15.2°, 17.0°, 17.7°, 18.2°, 18.6°, 19.3°, 19.8°, 20.2°, 21.4°, 22.5°, and 24.1°. In other aspects, crystal Form C is characterized by an XRPD substantially similar to FIG. 4. In other aspects, crystalline Form C is characterized by a Thermogravimetric analysis (TGA) curve with a step weight loss of 6.6% up to 150° C., or a Differential Scanning calorimetry (DSC) curve with three endotherms at 74.9° C. and 168.8° C. (onset temperatures) and 93.3° C. (peak temperature), or both, wherein the crystalline Form C may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form C is characterized by a Thermogravimetric analysis (TGA) or Differential Scanning calorimetry (DSC) substantially similar to FIG. 5, wherein the crystalline Form C may also comprise XRPD peaks at 2Θ angles selected from those above.

Also provided herein is a crystalline Form D of a compound having the structural formula:

In one aspect, crystalline Form D is characterized by at least three x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 17.4°, 21.3°, 22.2°, and 24.8°. In other aspects, crystalline Form D is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 17.4°, 21.3°, 22.2°, and 24.8°. In other aspects, crystalline Form D is characterized by at least five x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 17.4°, 21.3°, 22.2°, and 24.8°. In other aspects, crystalline Form D is characterized by at least six x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 17.4°, 21.3°, 22.2°, and 24.8°. In other aspects, crystalline Form D is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 17.4°, 21.3°, 22.2°, and 24.8°. In other aspects, crystalline Form D is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 4.9°, 9.9°, 11.1°, 14.8°, 17.4°, 21.3°, 22.2°, 24.8°, and 30.1°. In other aspects, crystal Form D is characterized by an XRPD substantially similar to FIG. 6. In other aspects, crystalline Form D is characterized by a Thermogravimetric analysis (TGA) curve with a step weight loss of 8.8% up to 150° C., or a Differential Scanning calorimetry (DSC) curve with two endotherms at 69.8° C. and 169.7° C. (onset temperatures) and an exotherm at 88.5° C. (peak temperature), or both, wherein the crystalline Form D may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form D is characterized by a Thermogravimetric analysis (TGA) or Differential Scanning calorimetry (DSC) substantially similar to FIG. 7, wherein the crystalline Form D may also comprise XRPD peaks at 2Θ angles selected from those above.

Also provided herein is a crystalline Form E of a compound having the structural formula:

In one aspect, crystalline Form E is characterized by at least three x-ray powder diffraction peaks at 2Θ angles selected from 8.0°, 16.0°, 16.8°, 17.1°, 18.8°, 19.3°, and 23.4°. In other aspects, crystalline Form E is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 8.0°, 16.0°, 16.8°, 17.1°, 18.8°, 19.3°, and 23.4°. In other aspects, crystalline Form E is characterized by at least five x-ray powder diffraction peaks at 2Θ angles selected from 8.0°, 16.0°, 16.8°, 17.1°, 18.8°, 19.3°, and 23.4°. In other aspects, crystalline Form E is characterized by at least six x-ray powder diffraction peaks at 2Θ angles selected from 8.0°, 16.0°, 16.8°, 17.1°, 18.8°, 19.3°, and 23.4°. In other aspects, crystalline Form E is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 8.0°, 16.0°, 16.8°, 17.1°, 18.8°, 19.3°, and 23.4°. In other aspects, crystalline Form E is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 3.2°, 8.0°, 9.6°, 11.0°, 12.1°, 12.6°, 14.1°, 14.7°, 16.0°, 16.8°, 17.1°, 17.7°, 18.8°, 19.3°, 19.6°, 20.1°, 21.5°, 21.7°, 22.2°, 22.4°, 23.4°, 24.3°, 25.5°, 26.9°, 27.8°, 28.3°, 29.3°, and 31.4°. In other aspects, crystalline Form E is characterized by an XRPD substantially similar to FIG. 8. In other aspects, crystalline Form E is characterized by a Thermogravimetric analysis (TGA) curve with a step weight loss of 8.1% up to 110° C., or a Differential Scanning calorimetry (DSC) curve two endotherms at 91.0° C. and 170.5° C. (onset temperatures), or both, wherein the crystalline Form E may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form E is characterized by a Thermogravimetric analysis (TGA) or Differential Scanning calorimetry (DSC) substantially similar to FIG. 9, wherein the crystalline Form E may also comprise XRPD peaks at 2Θ angles selected from those above.

Also provided herein is a crystalline Form F of a compound having the structural formula:

In one aspect, crystalline Form F is characterized by at least three x-ray powder diffraction peaks at 2Θ angles selected from 7.1°, 17.9°, 18.8°, 21.4°, 22.1°, 24.4°, and 28.8°. In other aspects, crystalline Form F is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 7.1°, 17.9°, 18.8°, 21.4°, 22.1°, 24.4°, and 28.8°. In other aspects, crystalline Form F is characterized by at least five x-ray powder diffraction peaks at 2Θ angles selected from 7.1°, 17.9°, 18.8°, 21.4°, 22.1°, 24.4°, and 28.8°. In other aspects, crystalline Form F is characterized by at least six x-ray powder diffraction peaks at 2Θ angles selected from 7.1°, 17.9°, 18.8°, 21.4°, 22.1°, 24.4°, and 28.8°. In other aspects, crystalline Form F is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.1°, 17.9°, 18.8°, 21.4°, 22.1°, 24.4°, and 28.8°. In other aspects, crystalline Form F is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 3.4°, 7.1°, 10.2°, 12.2°, 16.1°, 17.5°, 17.9°, 18.8°, 19.3°, 20.5°, 21.4°, 22.2°, 23.0°, 23.4°, 24.4°, 24.9°, 25.2°, 27.0°, 28.8°, 29.3°, 30.7°, and 32.1°. In other aspects, crystalline Form F is characterized by an XRPD substantially similar to FIG. 10. In other aspects, crystalline Form F is characterized by a Thermogravimetric analysis (TGA) curve with a step weight loss of 18.1% up to 150° C., or a Differential Scanning calorimetry (DSC) curve with two endotherms at 75.7° C. and 168.3° C. (onset temperatures) and an exotherm at 86.9° C. (peak temperature), or both, wherein the crystalline Form F may also comprise XRPD peaks at 2Θ angles selected from those above. In other aspects, crystalline Form F is characterized by a Thermogravimetric analysis (TGA) or Differential Scanning calorimetry (DSC) substantially similar to FIG. 11, wherein the crystalline Form F may also comprise XRPD peaks at 2Θ angles selected from those above.

Also provided herein is an amorphous form of a compound having the structural formula:

3. Uses, Formulation and Administration

The crystalline and amorphous forms described herein and compositions thereof are useful in treating diseases and/or disorders associated with the activity of potassium channels. Such diseases and/or disorders include e.g., neurodegenerative and neurological conditions (e.g., Parkinson's disease, tremors, Alzheimer's disease, dementia, amyotrophic lateral sclerosis (ALS) ataxia, anxiety, depression, mood disorders, memory and attention deficits, bipolar disorder, psychosis, schizophrenia, traumatic brain injury, and narcolepsy), heart disease and related conditions (e.g., ischaemic heart disease, coronary heart disease, angina pectoris, and coronary artery spasms), metabolic disease and bladder diseases (e.g., bladder spasms, urinary incontinence, bladder outflow obstruction, gastrointestinal dysfunction, irritable bowel syndrome, and diabetes), withdrawal symptoms associated with termination of addiction, and other conditions associated with the modulation of potassium channels such as e.g., respiratory diseases, epilepsy, convulsions, seizures, absence seizures, vascular spasms, renal disorders (e.g., polycystic kidney disease), erectile dysfunction, secretory diarrhoea, ischaemia, cerebral ischaemia, dysmenorrhea, Reynaud's disease, intermittent claudication, Sjorgren's syndrome, arrhythmia, hypertension, myotonic muscle dystrophia, spasticity, xerostomi, hyperinsulinemia, premature labor, baldness, cancer, immune suppression, migraine and pain.

The present disclosure also provides a method of modulating the activity of a potassium channel in a subject comprising the step of administering a crystalline or amorphous form or composition described herein. In another aspect, the present disclosure provides a method of positively modulating a SK2 channel in a cell comprising the step of contacting the cell with a crystalline or amorphous form or composition described herein.

In one aspect, the provided crystalline and amorphous forms described herein and compositions thereof are used to treat tremors. Tremors include, but are not limited to rest, active, postural, kinetic, intention, task specific, and idiopathic tremors. In one aspect, the provided crystalline and amorphous forms described herein and compositions thereof are used to treat postural and active tremors. Examples of postural and/or active tremors include essential tremor, drug-induced parkinsonism, neuropathic tremor, and tremors induced from toxins (e.g., alcohol withdrawal or from exposure to heavy metals). In one aspect, the provided crystalline and amorphous forms described herein and compositions thereof are used to treat essential tremor.

The present disclosure further provides a method of treating essential tremor in a subject comprising the step of administering a crystalline or amorphous form or composition described herein.

In some aspects, the crystalline and amorphous forms described herein and compositions thereof are useful in treating a disease or condition selected from a neurodegenerative disease, dementia, heart disease, withdrawal symptoms associated with termination of addiction, metabolic disease, and bladder disease. In some aspects, the crystalline and amorphous forms described herein and compositions thereof are useful in treating a disease or condition selected from ataxia, dystonia, Parkinson's disease, ischemia, traumatic brain injury, amyotrophic lateral sclerosis, hypertension, atherosclerosis, diabetes, arrhythmia, over-active bladder, and withdrawal symptoms caused by the termination of abuse of alcohol and other drugs of abuse. In some aspects, the crystalline and amorphous forms described herein and compositions thereof are useful in treating ataxia. In some aspects, the crystalline and amorphous forms described herein and compositions thereof are useful in treating spinocerebellar ataxia.

Also provided are pharmaceutically acceptable compositions comprising one or more of the disclosed crystalline or amorphous forms described herein; and a pharmaceutically acceptable carrier. These compositions can be used to treat one or more of the diseases and conditions described above.

Use of a disclosed crystalline or amorphous form described herein in the manufacture of a medicament for treating a disease or condition recited herein is also provided.

A crystalline or amorphous form described herein for use in treating a disease or condition recited herein is also provided.

Compositions may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Liquid dosage forms, injectable preparations, solid dispersion forms, and dosage forms for topical or transdermal administration of a crystalline or amorphous form are included herein.

The amount of provided crystalline or amorphous forms that may be combined with carrier materials to produce a composition in a single dosage form will vary depending upon the patient to be treated and the particular mode of administration. In some embodiments, provided compositions may be formulated so that a dosage of between 0.01-100 mg/kg body weight/day of the provided compound, such as e.g., 0.1-100 mg/kg body weight/day, can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the particular disease being treated. The amount of a provided crystalline and amorphous form in the composition will also depend upon the particular crystalline and amorphous form in the composition.

EXEMPLIFICATION

As depicted in the Examples below, crystalline and amorphous forms are prepared according to the following general procedures.

Abbreviations
Abbreviation Solvent
MeOH         Methanol
EtOH         Ethanol
IPA          Isopropyl alcohol
MIBK         4-Methyl-2-pentanone
EtOAc        Ethyl acetate
IPAc         Isopropyl acetate
MTBE         Methyl tert-butyl ether
THF          Tetrahydrofuran
2-MeTHF      2-Methyltetrahydrofuran
DCM          Dichloromethane
ACN          Acetonitrile
DMSO         Dimethylsulfoxide
DMAc         N,N-Dimethylacetamide
NMP          1-Methyl-2-pyrrolidone

Units
Full Name              Abbreviation
Celsius                C.
Degrees                °
Equivalents            eq.
Gram                   g
Hour                   hr
Kelvin                 K
Liters                 L
Milligrams             mg
Milliliters            mL
Minute                 min
Resolutions Per Minute rpm
Room temperature       RT
Second                 sec
volume                 vol.
Volume ratio           v/v
Watt                   W
Weight                 wt.
Weight Percentage      wt. %

1. Analytical Techniques

X-Ray Powder Diffraction (XRPD)

For XRPD analysis, a PANalytical Empyrean X-ray powder diffract meter was used. The XRPD parameters used are listed in Table 1.

TABLE 1
Parameters	Empyrean	X′ Pert3
X-Ray wavelength	Cu, kα;	Cu, kα;
	Kα1 (Å): 1.540598	Kα1 (Å): 1.540598
	Kα2 (Å): 1.544426	Kα2 (Å): 1.544426
	intensity ratio	intensity ratio
	Kα2/Kα1: 0.50	Kα2/Kα1: 0.50
X-Ray tube setting	45 kV, 40 mA	45 kV, 40 mA
Divergence slit	Automatic	Automatic
Scan mode	Continuous	Continuous
Scan range (°2TH)	3°~40°	3°~40°
Step size (°2TH)	0.0167	0.0263
Scan step time (s)	18	50
Test time (s)	5 min 30 s	5 min 04 s

Thermogravimetric Analysis (TGA) and Differential Scanning Calorimetry (DSC)

TGA data were collected using a TA Q500/Q5000 TGA from TA Instruments. DSC was performed using a TA Q200/Q2000 DSC from TA Instruments. Detailed parameters used are listed in Table 2.

TABLE 2
Parameters	TGA	DSC
Method	Ramp	Ramp
Sample pan	Aluminum, open	Aluminum, crimped
Temperature	RT-desired temperature	25° C.-desired temperature
Heating rate	10° C./min	10° C./min
Purge gas	N2	N2

Dynamic Vapor Sorption (DVS)

DVS was measured via a SMS (Surface Measurement Systems) DVS Intrinsic. The relative humidity at 25° C. were calibrated against deliquescence point of LiCl, Mg(NO₃)₂ and KCl. Parameters for DVS test were listed in Table 3.

TABLE 3
Parameters                 DVS
Temperature                25° C.
Sample size                10~20 mg
Gas and flow rate          N2, 200 mL/min
dm/dt                      0.002%/min
MM. dm/dtstabilityduration 10 min
Max. equilibrium time      180 min
RH range                   0% RH to 95% RH
RH step size               10% RH from 0% RH to 90% RH
                           5% RH from 90% RH to 95% RH

¹HNMR

¹HNMR was collected on Bruker 400M NMR Spectrometer using DMSO-d6.

Single Crystal X-Ray Diffraction (SCXRD)

The SCXRD data were collected at 150 K using Agilent SuperNova (Dual, Cu at zero, Eos) diffractometer (Cu/Kα radiation, λ=1.54178 Å). The parameters used are listed in Table 4. The microscopic picture was captured using Shanghai Cewei PXS9-T stereo microscope.

TABLE 4
Device                  Agilent SuperNova
X-Ray sources generator SuperNova Microfocus X-ray Source
                        (Cu/kα: 1.54178 Å) 50 KV, 0.8 mA
Detector                Eos CCD detector
                        (Detector resolution: 16.0450 pixels mm−1)
Goniometer              Four-circle Kappa geometry goniometer
Low Temperature Devices Oxford Cryosystems
Software                CrysAlisPro (version: 1.171.38.41)

Single Crystal Structure Determination

The structure was solved with the ShelXS structure solution program using Direct Methods (Sheldrick G M Acta Cryst 2008, A64:112-122) and refined with ShelXL refinement package using full-matrix least-squares on F contained in OLEX2 (Sheldrick G M Acta Cryst 2015, C71:3-8; Dolomanov O V, et al. J Appl Cryst 2006, 42:339-341). The calculated XRPD pattern was obtained by Mercury (Macrae C F, et al. J Appl Cryst 2006, 39:453-457) and the crystal structure representations were generated by Diamond (Brandenburg K DIAMOND 1999, Crystal Impact GbR, Bonn, Germany). The ORTEP (Oak Ridge Thermal Ellipsoid Plot) drawing was generated by ORTEP-III (Farrugia L J J Appl Cryst 2012, 45:849-854). The Platon program version: 21116 (Analysis of Potential Hydrogen Bonds with d(D . . . A)<R(D)+R(A)+0.50, d(H . . . A)<R(H)+R(A)−0.12 Angle, D-H . . . A>100.0 Degree) was used to calculate the classic hydrogen bonds in the crystal structure.

2. Polymorph Screening

The starting material for the following screens was obtained following the procedure set forth in Example 1 of U.S. Pat. No. 9,975,886, the entire contents of which are incorporated herein by reference, except that the material used for polymorph screening was isolated by trituration with alcoholic solvent instead of column chromatography. The XRPD of this material is shown in FIG. 12 as “Form A” and was determined to be a mixture of approximately 96% Form B and approximately 4% of one or more additional forms having certain characteristic peaks of Form C. These percentages are based on the relative intensity differences between the Form B representative peak at 7.2 and the Form C representative peak at 8.5 for the starting material (i.e., Form A) in FIG. 12. Estimating a baseline adjustment of about −1020 counts, the intensity for the Form B representative peak at 7.2 is about 8571 counts (about 9591-about 1020), whereas the intensity for the Form C representative peak at 8.5 is about 408 counts (about 1428-about 1020).

Solubility measurements for this mixture are provided in Table 5 and were obtained using the following procedure: approximately 2 mg of Form A was added into a 3-mL glass vial, a solvent of interest was then added into the vial until the solids were dissolved or a total volume of 1 mL was reached.

TABLE 5
Solvent     Solubility (mg/mL)
MeOH        S > 44.0
EtOH        S > 42.0
IPA         6.3 < S < 19.0
acetone     S > 42.0
MIBK        20.0 < S < 40.0
EtOAc       21.0 < S < 42.0
IPAc        6.3 < S < 19.0
MTBE        2.0 < S < 6.7
THF         S > 38.0
2-MeTHF     S > 40.0
1,4-dioxane S > 42.0
ACN         S > 40.0
DCM         S > 40.0
CHC13       S > 44.0
n-heptane   S < 2.1
toluene     6.7 < S < 20.0
DMSO        S > 38.0
DMAc        S > 40.0
NMP         S > 42.0
H2O         S < 2.1

Using Form A as the starting material, preliminary polymorph screening experiments were performed. The methods used and identified crystalline forms are summarized in Table 6 and further detailed below. Form E was observed in-situ after either purging hydrate Form D sample under N₂ at 30° C. for 30 min or heating to 105° C.

TABLE 6
Method                          Single Crystal Forms Identified
Anti-solvent addition           Form B,
Solid vapor diffusion           Form B, Form E, amorphous Form
Slow evaporation                Form B,
Slow cooling                    Form B, Form C
Slurry at RT                    Form B
Slurry at 50° C.                Form B
Liquid vapor diffusion          Form B, Form C
Polymer induced crystallization Form B, Form D

Anti-Solvent Addition

A total of 14 anti-solvent addition experiments were carried out, using the solvent and anti-solvent systems listed in Table 7. About 20 mg of Form A was dissolved in 0.2˜2.0 mL solvent to obtain a clear solution; the clear solution was magnetically stirred, followed by the step-wise addition of 0.1 mL anti-solvent until a precipitate appeared or the total amount of the anti-solvent reached 10.0 mL. The precipitate was then isolated for XRPD analysis.

TABLE 7
Solvent	Anti-solvent	Single Solid Form
EtOH	n-heptane	Form B*
MIBK		Form B
IPAc		Form B
2-MeTHF		Form B
1,4-dioxane		Form B
DCM		Form B
toluene		Form B
MeOH	H2O	Form B
acetone		Form B
1,4-dioxane		Form B
ACN		Form B
DMSO		Form B
NMP		Form B
*clear solution was obtained and then transferred to slurry at 5° C.

Solid Vapor Diffusion

Solid vapor diffusion experiments were conducted under 12 solvent conditions, as shown in Table 8. For each solvent condition, about 15 mg of Form A was weighed into a 3-mL vial, which was placed into a 20-mL vial with 4 mL volatile solvent. The 20-mL vial was sealed with a cap and kept at RT for 8 days allowing the solvent vapor to interact with Form A. The solids obtained were then analyzed by XRPD.

TABLE 8
Solvent     Single Solid Form
H2O         Form B
DCM         Amorphous Form*
EtOH        Form B
MeOH        Form E
ACN         Form B
acetone     Form B
DMSO        Form B
EtOAc       Form B*
1,4-dioxane Form B
IPA         Form B
*solid dissolved completely and then transferred to evaporate at RT.

Slow Evaporation

Slow evaporation experiments were performed under 12 different solvent conditions, as shown in Table 9. About 20 mg of Form A was dissolved in 0.4˜3.0 mL solvent in a 3-mL glass vial. If not dissolved completely, suspensions were filtered using a PTFE membrane (pore size of 0.45 μm) and the filtrates would be used instead for the follow-up steps. The visually clear solutions were subjected to evaporation at RT with vials sealed by Parafilm®. The solids yielded were then isolated for XRPD analysis.

TABLE 9
Solvent (v/v)       Single Solid Form
MeOH                Form B
acetone             Form B
IPAc                Form B
THF                 Form B
CHCl3               Form B
EtOAc/MTBE (1:1)    Form B
EtOH/2-MeTHF (1:1)  Form B
DCM/MIBK (1:1)      Form B
THF/n-heptane (1:1) Form B
ACN/H2O (1:1)       Form B

Slow Cooling

Slow cooling experiments were conducted under 10 different solvent conditions, as shown in Table 10. For each solvent condition, about 20 mg Form A was suspended in 0.5-1.0 mL solvent in a 3-mL glass vial at RT. The suspension was then heated to 50° C., equilibrated for about 2 hrs and filtered into a new vial using a PTFE membrane (pore size of 0.45 μm). The filtrate was slowly cooled down to 5° C. at a rate of 0.1° C./min. The solids were then obtained and kept isothermally at 5° C. before being isolated for XRPD analysis.

TABLE 10
Solvent (v/v)           Single Solid Form
IPA                     Form C
IPAc                    Form B
MTBE                    Form B
toluene                 Form B
IPA/MTBE (1:1)          Form C
IPAc/toluene (1:1)      Form B
acetone/n-heptane (1:1) Form B
ACN/H2O (1:1)           Form B
THF/n-heptane (1:2)     Form B

Slurry at RT

Slurry conversion experiments were conducted at RT under 20 different solvent systems, as shown in Table 11. About 20 mg Form A was suspended in 0.3 mL solvent in a 1.5-mL glass vial. After the suspension was stirred magnetically for 4 days at RT, the remaining solids were isolated for XRPD analysis.

TABLE 11
Solvent (v/v)               Single Solid Form
IPA                         Form B
IPAc                        Form B
MTBE                        Form B
n-heptane                   Form B
toluene                     Form B
H2O                         Form B
MeOH/H2O (1:9)              Form B
EtOAc/n-heptane (1:9)       Form B
THF/H2O (1:4)               Form B
DCM/n-heptane (1:9)         Form B
IPAc/MTBE (1:1)             Form B
IPA/toluene (1:1)           Form B
MIBK/n-heptane (1:9)        Form B
DMAc/H2O (1:4)              Form B
acetone/n-heptane (1:9)     Form B
1,4-dioxane/n-heptane (1:9) Form B
IPA/H2O (aw~0.2, 98:2)      Form B
IPA/H2O (aw~0.4, 96:4)      Form B
IPA/H2O (aw~0.6, 92:8)      Form B
IPA/H2O (aw~0.8, 85:15)     Form B

Slurry at 50° C.

Slurry conversion experiments were conducted at 50° C. under 14 different solvent systems, as shown in Table 12. About 25 mg Form A was suspended in 0.3 mL solvent in a 1.5-mL glass vial. After the suspension was stirred for about 4 days at 50° C., the remaining solids were isolated for XRPD analysis.

TABLE 12
Summary of slurry conversion experiments at 50° C.
Solvent (v/v)           Single Solid Form
IPA                     Form B
IPAc                    Form B
MTBE                    Form B
n-heptane               Form B
toluene                 Form B
H2O                     Form B
IPAc/toluene (1:1)      Form B
IPA/MTBE (1:1)          Form B
EtOH/H2O (1:9)          Form B
MIBK/n-heptane (1:9)    Form B
toluene/n-heptane (1:1) Form B
ACN/H2O (1:9)           Form B
CHCl3/n-heptane (1:9)   Form B
THF/H2O (1:9)           Form B

Liquid Vapor Diffusion

12 liquid vapor diffusion experiments were conducted, using the solvent and anti-solvent systems listed in Table 13. About 20 mg Form A was dissolved in 0.2˜2.0 mL appropriate solvent to obtain a clear solution in a 3-mL vial. This solution was then placed into a 20-mL vial with 4 mL volatile solvents. The 20-mL vial was sealed with a cap and kept at RT allowing sufficient time for organic vapor to interact with the solution. After the slow cooling process, clear solutions were obtained for the IPA and 2-MeTHF solvent systems, and solids were obtained for all other solvent systems. The solids were then isolated for XRPD analysis. The clear solutions were evaporated to dryness at RT to obtain the solids for XRPD analysis.

TABLE 13
Solvent	Anti-solvent	Single Solid Form
IPA	n-heptane	Form C*
MIBK		Form B
EtOAc		Form B
MTBE		Form B
CHCl3		Form B
toluene		Form B
EtOH	H2O	Form B
acetone		Form B
DMAc		Form B
1,4-dioxane		Form B
ACN		Form B
*Clear solution was obtained and transferred to evaporate at RT.

Polymer Induced Crystallization

As shown in Table 14, polymer induced crystallization experiments were performed with two sets of polymer mixtures in six solvents, wherein polymer mixture A consists of polyvinyl pyrrolidone (PVP), polyvinyl alcohol (PVA), polyvinylchloride (PVC), polyvinyl acetate (PVAC), hypromellose (HPMC), methyl cellulose (MC) in a mass ratio of 1:1:1:1:1:1; and polymer mixture B consists of polycaprolactone (PCL), polyethylene glycol (PEG), poly(methyl methacrylate) (PMMA) sodium alginate (SA), and hydroxyethyl cellulose (HEC) in a mass ratio of 1:1:1:1:1. For each experiment, about 20 mg Form A was dissolved in 0.4˜3.0 mL appropriate solvent to obtain a clear solution in a 3-mL vial. About 2 mg of a polymer mixture was added into 3-mL glass vial. All the samples were subjected to evaporate at RT to induce precipitation. The solids were isolated for XRPD analysis.

TABLE 14
Solvent (v/v)	Polymer	Single Solid Form
EtOH	Polymer	Form D
MIBK/n-heptane (1:4)	mixture A	Form B
EtOAc/1,4-dioxane (1:1)		Form B
CHCl3	Polymer	Form B
IPAc/ACN (1:1)	mixture B	Form B
2-MeTHF/toluene (1:1)		Form B

Polymorph Screening Using the Amorphous Form as the Starting Material

The amorphous Form of Compound 1 was prepared according to the procedures set forth in Example 6 below. Using the amorphous Form as the starting material, 10 additional polymorph screening experiments, referred to as slurry conversion experiments, were conducted at RT or 50° C. in different solvent systems, as shown in Table 15. About 10 mg of the amorphous Form was suspended in 0.2˜0.3 mL of solvent. After the suspension was stirred magnetically (˜1000 rpm) at RT or 50° C., the remaining solids were isolated for XRPD analysis.

TABLE 15
		Single Solid
Solvent (v/v)	Temperature	Form
H2O	RT	Form B
n-heptane		Form B
MTBE		Form B
acetone/H2O (1:9)		Form B
CHCl3/n-heptane (1:9)		Form B
H2O	50° C.	Form B
n-heptane		Form B
IPA/H2O (1:4)		Form B
EtOAc/n-heptane (1:9)		Form B
MIBK/toluene (1:9)		Form B

3. Preparation of Various Forms

Example 1: Preparation of Single Crystalline Form B

Form B was obtained via heating Form A to 140° C. and then cooled to RT. Form B showed a strong, unique XRPD pattern with sharp peaks and a flat baseline, indicative of a crystalline material (see FIG. 1 and Table 16). Form B showed a TGA curve with a 0.1% weight loss up to 150° C. and a DSC curve with a sharp endotherm at 170.6° C. (onset temperature) (FIG. 2). The low TGA weight loss and the sole sharp DSC endotherm suggested that Form B was an anhydrate.

The crystal structure of Form B at the atomic and molecular level was determined by SCXRD (see FIG. 3), using the sample obtained by slow evaporation from ACN/H2O (1:1, v/v) solution at RT. The SCXRD analysis revealed that the crystal adopted the Monoclinic crystal system and P2₁/c space group with a=15.1466(2) Å, b=24.6770(4) Å, c=9.87220(17) Å, α=90°, β=94.4397(15)°, γ=90°, V=3678.89(10) ų. The asymmetric unit of the Form B single crystal (FIG. 3A) consisted of two Compound 1 molecules (FIG. 3B), suggesting the crystal was an anhydrate. The unit cell of the Form B single crystal consisted of eight Compound 1 molecules (FIG. 3C). FIG. 3D showed the classic hydrogen bonds and FIG. 3E showed the 3D packing of the unit cell viewed along c-axis in the Form B single crystal structure. The calculated XRPD from the single crystal structure data was consistent with the experimental XRPD of Form B (FIG. 3F).

TABLE 16
XRPD of single crystalline Form B
Pos. [°2Th.]	d-spacing [Å]	Rel. Int. [%]
7.156134	12.35308	60.27
9.227475	9.58425	10.66
10.886250	8.12731	15.13
11.526900	7.67700	1.49
12.177290	7.26840	1.69
13.094060	6.76149	15.96
13.671450	6.47721	3.21
13.960340	6.34381	4.14
14.600230	6.06719	10.17
15.476650	5.72553	100.00
16.888730	5.24986	5.05
17.665540	5.02072	21.50
18.364500	4.83118	8.57
18.878580	4.70077	30.47
19.146590	4.63557	6.78
20.050540	4.42857	26.55
20.621050	4.30732	11.03
20.926090	4.24522	8.78
21.425230	4.14743	13.13
21.995030	4.04126	16.58
22.304330	3.98592	21.56
23.156060	3.84120	37.18
23.974810	3.71184	7.00
24.534800	3.62838	6.46
25.202700	3.53371	0.89
25.891670	3.44122	2.86
26.353470	3.38196	1.15
27.119010	3.28821	3.15
28.121280	3.17325	5.38
28.886390	3.09092	3.71
29.716040	3.00648	4.36
33.910100	2.64362	2.47
34.673120	2.58718	1.22
35.183730	2.55079	1.34
37.334850	2.40862	1.78
38.910120	2.31466	0.72

GMP Manufacture of Single Crystalline Form B Form

Form A (6.53 Kg) was dissolved in 2-MeTHF and the solution was heated to 70° C. Over the course of approximately 80 minutes, heptane (52.79 L, 35.90 kg, 8 volumes) was slowly charged to the reactor via an addition funnel. Agitation was momentarily paused, allowing water that had separated out of solution to settle on the bottom of the reactor. The water was drained (1.04 L), and agitation resumed for not less than 30 minutes. Over a span of 3 hours, the reactor jacket temperature ramped down (linear) from 70° C. to 60° C. During this period, the crystallization of solids was observed (internal process temperature was approximately 67° C.). The temperature was held at 60° C. for a duration of 3 hours, followed by a linear ramped-down over a 3-hour period from 60° C. to 20° C. The solids were transferred from the reactor to a 24″ polypropylene table-top filter funnel. The product was washed two times with polish-filtered heptane (32.69 L, 22.23 kg, 5 volumes) for a total wash of 10 volumes. The desired Form B product was transferred to Teflon lined drying trays, and dried under vacuum at 50° C. with a nitrogen bleed, until constant mass was achieved. Drying occurred over the course of 4 days, yielding the desired product, Form B, as a light-yellow solid in 82% yield.

Example 2: Preparation of Single Crystalline Form C

Form C was obtained by slow evaporation from an IPA solution at RT. Form C showed a strong, unique XRPD pattern with sharp peaks and a relatively flat baseline, indicative of a crystalline material (see FIG. 4 and Table 17). Form C showed a TGA curve with a step weight loss of 6.6% up to 150° C., as well as a DSC curve with three endotherms at 74.9° C. and 168.8° C. (onset temperatures) and 93.3° C. (peak temperature) (FIG. 5). Form C was found to be IPA solvate that converted to anhydrate Form B after heated to 110° C. or stored at RT for 8 days, by ¹H solution NMR and XRPD analyses.

TABLE 17
XRPD of single crystalline Form C
Pos. [°2Th.]	d-spacing [Å]	Rel. Int. [%]
4.229768	20.89073	34.39
4.544583	19.44426	4.34
8.464246	10.44668	82.24
9.096947	9.72147	1.17
9.698006	9.12027	4.74
10.416890	8.49242	5.24
13.083840	6.76675	1.34
13.746750	6.44190	3.71
15.229290	5.81797	0.90
17.041950	5.20300	100.00
17.686770	5.01474	2.32
18.244950	4.86256	2.38
18.573650	4.77725	3.89
19.278020	4.60426	1.56
19.815810	4.48050	1.82
20.151740	4.40656	9.52
21.367220	4.15856	59.09
22.459880	3.95866	4.15
24.109100	3.69147	2.07

Example 3: Preparation of Single Crystalline Form D

Form D was obtained via slow evaporation from EtOH solution at RT. Form D showed a strong, unique XRPD pattern with sharp peaks and a relatively flat baseline, indicative of a crystalline material (see FIG. 7 and Table 18). Form D showed a TGA curve with a step weight loss of 8.8% up to 150° C. as well as a DSC curve with two endotherms at 69.8° C. and 169.7° C. (onset temperatures) and an exotherm at 88.5° C. (peak temperature) (FIG. 8). Form D was found to be EtOH solvate that converted to anhydrate Form B after stored at RT for a month, by ¹H solution NMR and XRPD analyses.

TABLE 18
XRPD of single crystalline Form D
Pos. [°2Th.]	d-spacing [Å]	Rel. Int. [%]
4.898084	18.04170	22.81
9.870212	8.96153	47.21
11.099930	7.97133	52.87
14.809040	5.98211	13.93
17.394970	5.09820	100.00
21.310910	4.16942	31.74
22.244260	3.99654	68.72
24.792990	3.59117	14.08
30.118660	2.96721	8.12

Example 4: Preparation of Single Crystalline Form E

Form E sample was obtained via placing the Form A starting material in a chamber full of volatile MeOH solvent at RT for 8 days. Form E showed a strong, unique XRPD pattern with sharp peaks and a relatively flat baseline, indicative of a crystalline material (see FIG. 8 and Table 19). Form E showed a TGA curve with a step weight loss of 8.1% up to 110° C., as well as a DSC curve with two endotherms at 91.0° C. and 170.5° C. (onset temperatures) (FIG. 9). Form E was found to be MeOH solvate that converted to anhydrate Form B after heated to 130° C., by ¹H solution NMR and XRPD analyses.

TABLE 19
XRPD of single crystalline Form E
Pos. [°2Th.]	d-spacing [Å]	Rel. Int. [%]
3.184209	27.74763	17.23
7.996826	11.05621	77.65
9.638795	9.17616	29.59
11.045390	8.01056	41.70
12.115180	7.30552	18.93
12.591690	7.03010	50.90
14.056210	6.30076	24.99
14.688860	6.03078	22.42
15.994650	5.54125	86.61
16.815980	5.27241	70.23
17.113930	5.18128	57.68
17.670550	5.01931	6.61
18.797420	4.72088	100.00
19.340050	4.58963	82.07
19.630910	4.52228	15.29
20.101930	4.41737	10.46
21.535870	4.12637	38.06
21.749990	4.08623	41.36
22.153240	4.01276	31.02
22.420710	3.96549	36.04
23.381110	3.80473	84.99
24.258590	3.66906	9.81
25.451220	3.49977	9.70
26.896600	3.31489	8.26
27.818180	3.20713	15.04
28.346960	3.14850	14.69
29.276680	3.05060	15.02
31.408110	2.84827	5.19

Example 5: Preparation of Single Crystalline Form F

Form F was obtained by evaporation of DCM solution (100 mg scale) at RT for 1 day. Form F showed a strong, unique XRPD pattern with sharp peaks and a relatively flat baseline, indicative of a crystalline material (see FIG. 10 and Table 20). Form F showed a TGA curve with a step weight loss of 18.1% up to 150° C., as well as a DSC curve with two endotherms at 75.7° C. and 168.3° C. (onset temperatures) and an exotherm at 86.9° C. (peak temperature) (FIG. 11). Form F was found to be DCM solvate that converted to anhydrate Form B after heated to 120° C., by ¹H solution NMR and XRPD analyses.

TABLE 20
XRPD of single crystalline Form F
Pos. [°2Th.]	d-spacing [Å]	Rel. Int. [%]
3.398548	25.99811	6.08
7.084709	12.47746	59.40
10.164230	8.70296	3.25
12.247130	7.22710	3.22
16.098370	5.50578	1.73
17.451150	5.08192	7.97
17.940350	4.94443	9.62
18.813090	4.71698	10.78
19.260910	4.60831	3.12
20.459110	4.34105	7.07
21.387790	4.15461	100.00
22.149230	4.01348	12.73
22.973100	3.87137	3.37
23.430970	3.79675	3.58
24.402940	3.64768	11.80
24.918290	3.57340	6.59
25.226130	3.53048	7.06
26.994900	3.30304	3.89
28.822940	3.09758	9.85
29.317950	3.04640	2.24
30.744450	2.90822	1.17
32.083800	2.78981	1.13

Example 6: Preparation of the Amorphous Form of Compound 1

Two batches of the amorphous Form (50-mg and 200-mg scale) were prepared via evaporation of DCM solution of the Form A starting material at RT. For the 200-mg batch, 202.0 mg of Form A was weighed into a 3-mL vial followed by the addition of 1 mL of DCM to dissolve the solids. The solution was filtered using a PTFE membrane (pore size of 0.45 μm) and the filtrate was subject to evaporate in vacuum at 30° C. XRPD and mDSC results are shown in FIG. 13 and FIG. 14. No glass transition temperature (Tg) was observed for either batch of the amorphous form.

Claims

1. A single crystalline Form B of a compound having structural formula 1:

2. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by at least four x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°.

3. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by at least five x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°.

4. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by at least six x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°.

5. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 15.5°, 17.7°, 18.9°, 20.1°, 22.3°, and 23.2°.

6. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 9.2°, 10.9°, 13.1°, 14.6°, 15.5°, 17.7°, 18.9°, 20.1°, 20.6°, 21.4°, 22.0°, 22.3°, and 23.2°.

7. The single crystalline Form B of claim 1, wherein the crystalline form is characterized by x-ray powder diffraction peaks at 2Θ angles selected from 7.2°, 9.2°, 10.9°, 11.5°, 12.2°, 13.1°, 13.7°, 14.0°, 14.6°, 15.5°, 16.9°, 17.7°, 18.4°, 18.9°, 19.1°, 20.1°, 20.6°, 20.9°, 21.4°, 22.0°, 22.3°, 23.2°, 24.0°, 24.5°, 25.2°, 25.9°, 26.4°, 27.1°, 28.1°, 28.9°, 29.7°, 33.9°, 34.7°, 35.2°, 37.3°, and 38.9°.

8. A single crystalline Form C, single crystalline Form D, single crystalline Form E, or single crystalline Form F of a compound having structural formula 1:

9. A pharmaceutical composition comprising the crystalline Form B of claim 1; and a pharmaceutically acceptable carrier.

10. A method of treating a disease or condition responsive to modulation of the small conductance calcium-activated potassium channel (SK channel) in a subject comprising the step of administering to the subject a therapeutically effective amount of the crystalline Form B of claim 1.

11. A method of preparing the crystalline Form B of claim 1, said method comprising: dissolving Form A in 2-MeTHF at elevated temperature to form a solution;adding heptane to the solution; andreducing the temperature of the solution, thereby precipitating Form B.

12. A pharmaceutical composition comprising the crystalline Form C, Form D, Form E, or Form F of claim 8, and a pharmaceutically acceptable carrier.

13. A method of treating a disease or condition responsive to modulation of the small conductance calcium-activated potassium channel (SK channel) in a subject comprising the step of administering to the subject a therapeutically effective amount of the crystalline Form C, Form D, Form E, or Form F of claim 8.