PHENYLHEXANAMIDE

RELATED APPLICATIONS

[0001] This application claims priority to, and the benefit of, U.S. Provisional Application Nos. 63/164,266, filed March 22, 2021, and 63/164,286, filed March 22, 2021, the entire contents of each of which are incorporated herein by reference.

BACKGROUND

[0002] Mitochondrial dysfunction may contribute to various types of neurodegenerative diseases. Defective mitochondrial fusion or fission may be especially problematic in this regard, especially when imbalanced fusion and fission lead to mitochondrial fragmentation. Among the many neurodegenerative diseases in which mitochondrial dysfunction has been implicated include, for example, Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis (ALS), and Huntington’s disease.

[0003] Mitochondrial fusion is initiated by outer mitochondrial membrane-embedded mitofusin (MFN) proteins whose extra-organelle domains extend across cytosolic space to interact with counterparts on neighboring mitochondria. The physically linked organelles create oligomers of varying sizes. Mitofusins subsequently induce outer mitochondrial membrane fusion mediated by catalytic GTPase. Aberrant mitofusin activity is believed to be a primary contributor to mitochondrial-based neurodegenerative diseases. For these reasons, mitofusins are attractive targets for drug discovery.

[0004] There remains a need for new compounds that target mitofusins. The present disclosure addresses the need.

SUMMARY

[0005] In some aspects, the present disclosure provides a crystalline polymorph of a compound represented by the structure

N-(trans-4-hydroxycyclohexyl)-6-phenylhexanamide [0006] In some aspects, the present disclosure provides a method of preparing a compound described herein.

[0007] In some aspects, the present disclosure features a pharmaceutical composition comprising any compound described herein and a pharmaceutically acceptable excipient. [0008] In some aspects, the present disclosure features a method of treating diseases, disorders, or conditions, comprising administering to a subject in need thereof any compound described herein in a pharmaceutical composition.

[0009] In some aspects, the present disclosure features any compound described herein in a pharmaceutical composition for use for treating diseases, disorders, or conditions, comprising administering to a subject in need thereof.

[0010] In some aspects, the present disclosure features use of any compound described herein in a pharmaceutical composition in the manufacture of a medicament for treating diseases, disorders, or conditions, composing administering to a subject in need thereof.

[0011] In some aspects, the present disclosure features a method of activating mitofusin in a subject, comprising administering the compound or the pharmaceutical composition of any one of the preceding claims.

[0012] In some aspects, the present disclosure features any compound described herein in a pharmaceutical composition for use in activating mitofusin in a subject.

[0013] In some aspects, the present disclosure features use of the any compound described herein in a pharmaceutical composition in the manufacture of a medicament for activating mitofusin in a subject.

[0014] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the specification, the singular forms also include the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference. The references cited herein are not admitted to be prior art to the claimed invention. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods and examples are illustrative only and are not intended to be limiting. In the case of conflict between the chemical structures and names of the compounds disclosed herein, the chemical structures will control. [0015] Other features and advantages of the disclosure will be apparent from the following detailed description and claims.

BRIEF DESCRIPTION OF THE DRAWINGS [0016] The following figures are included to illustrate certain aspects of the present disclosure, and should not be viewed as exclusive embodiments. The subject matter disclosed is capable of considerable modifications, alterations, combinations, and equivalents in form and function, as will occur to one having ordinary skill in the art and having the benefit of this disclosure. [0017] FIG. 1 shows an illustrative x-ray powder diffraction pattern of as-produced N -(trans- 4-hydroxycyclohexyl)-5-phenylpentanamide (Polymorph 1).

[0018] FIG. 2 shows an illustrative polarized light microscopy image of as-produced N -(trans- 4-hydroxycyclohexyl)-5-phenylpentanamide (Polymorph 1).

[0019] FIG. 3 shows an illustrative x-ray powder diffraction pattern of Polymorph 2 of N- (trans-4-hydroxycyclohexy-5- phenylpentanamide

[0020] FIG. 4 shows an illustrative polarized light microscopy image of Polymorph 2 of N- (trans-4-hydroxycyclohexy-5- phenylpentanamide

[0021] FIG. 5 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 2 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide.

[0022] FIG. 6 shows an illustrative x-ray powder diffraction pattern of Polymorph 3 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0023] FIG. 7 shows an illustrative polarized light microscopy image of Polymorph 3 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0024] FIG. 8 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 3 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide.

[0025] FIG. 9 shows an illustrative x-ray powder diffraction pattern of Polymorph 4 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0026] FIG. 10 shows an illustrative polarized light microscopy image of Polymorph 4 ofN-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0027] FIG. 11 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 4 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide. [0028] FIG. 12 shows an illustrative x-ray powder diffraction pattern of Polymorph 5 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0029] FIG. 13 shows an illustrative polarized light microscopy image of Polymorph 5 of N-(trans-4-hydroxycyclohexy-5- phenylpentanamide

[0030] FIG. 14 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 5 of (trans-4-hydroxycyclohexy-5- phenylpentanamide.

[0031] FIG. 15 shows an overlay plot of the x-ray powder diffraction patterns of Polymorphs

1-5

DETAILED DESCRIPTION

[0032] Although several chemical entities have been found to target mitofusins, most have failed to display pharmacokinetic properties compatible with in vivo use. U.S. Patent Application Publication 2020/0345669 describes various N-(cycloalkyl or heterocycloalkyl) 6- phenylhexanamide compounds or structural variants thereof capable of activating mitofusins. 4-Hydroxycyclohexyl is a particularly effective substituted cycloalkyl in such mitofusin activators. U.S. Patent Application Publication 2020/0345668 describes stereoisomeric N-(4- hydroxycyclohexyl)-6-phenylhexanamide compounds or structural variants thereof that are particularly effective for promoting mitofusin activation. N-(trans-4-hydroxycyclohexy- 6 - phenylhexanamide is the active stereoisomer, whereas the corresponding cis-stereoisomer is inactive toward promoting mitofusin activation.

[0033] The present disclosure generally relates to mitofusin activation, and more specifically, compositions comprising a crystalline polymorph of N-(trans-4-hydroxycyclohexy- 6 - phenylhexanamide.

[0034] As discussed above, various N-(cycloalkyl or heterocycloalkyl)-6-phenylhexanamide compounds are potent mitofusin activators. N-(trans-4-hydroxycyclohexy- 6 - phenylhexanamide (Formula 1) is an example of a particularly potent mitofusin activator.

Formula 1 N-(trans-4-hydroxycyclohexy- 6 -phenylhexanamide [0035] Without wishing to be bound by theory, N-(trans-4-hydroxycyclohexy- 6 - phenylhexanamide is a mitofusin activator may exist as at least five different crystalline polymorphs. The polymorphs may be anhydrates. A crystalline polymorph formed above a phase-transition temperature of about 125°C may be characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 18.43° 2θ, as measured using Cu Kα radiation. A crystalline polymorph formed below a phase transition temperature of about 40°C may be characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 18.57° 2θ, as measured using Cu Kα radiation. Pharmaceutical compositions may be prepared from the polymorphs.

[0036] N-(trans-4-hydroxycyclohexy- 6 -phenylhexan aammiiddee may be obtained as a crystalline polymorph representing the most stable form produced above about 125°C. This polymorph is characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 18.43° 2θ, as measured using Cu Kα radiation. Herein, this polymorph formed at elevated temperatures is referred to as Polymorph 1, which may be obtained as an anhydrate. Polymorph 1 may also be formed through crystallization via antisolvent addition. Polymorph 1 may be converted into at least four additional crystalline polymorphs formed at lower temperatures under various conditions. These additional polymorphs, discussed hereinafter and referred to as Polymorphs 2-5 herein, each have distinctive x-ray powder diffraction patterns and may be obtained readily through manipulation of Polymorph 1 under various conditions. FIG. 15 shows an overlay plot of the x-ray powder diffraction patterns of Polymorphs 1-5.

[0037] Polymorph 1 exhibits additional x-ray powder diffraction peaks at the following approximate 2θ values, as measured using Cu Kα radiation: 17.70° and 17.86° (overlapping), 19.33° and 20.49°. Each of these peaks is present at a significant fraction of the intensity of the most intense peak of Polymorph 1. One or more additional peaks may be observed at the following approximate 2θ values, as measured using Cu Kα radiation: 9.19°, 10.27°, 11.71°, 13.55°, 15.62°, 21.90°, 26.76°, and 27.15°. These peaks are present at a considerably lower fraction of the intensity of the most intense peak of Polymorph 1.

[0038] Polymorph 1 may be converted into a second crystalline polymorph (Polymorph 2) characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 17.44° 2θ, as measured using Cu Kα radiation. In particular, Polymorph 1 may be converted to Polymorph 2 under thermal cycling conditions by transitioning between a temperature of 5°C and 50°C when slurried in various solvents. Polymorph 2 may form at a phase transition temperature of about 40°C to about 60° and be obtained as an anhydrate. In a non-limiting example, temperature cycling to produce Polymorph 2 may be conducted in a 1 :2 ethanol/heptane (vol.wol.) slurry of Polymorph 1, as well as in other solvents producing a slurry of Polymorph 1. Polymorph 2 may revert to Polymorph 1 upon heating above about 125°C.

[0039] Polymorph 2 exhibits additional x-ray powder diffraction peaks at the following approximate 2θ values, as measured using Cu Kα radiation: 8.72°, 17.99°, 18.46°, 18.83°, 19.51°, 19.98°, and 20.85°. Each of these peaks is present at a significant fraction of the intensity of the most intense peak of Polymorph 2. One or more additional peaks may be observed at the following approximate 2θ values, as measured using Cu Kα radiation: 11.51°, 13.24°, 14.03°, 15.50°, 15.86°, 16.18°, 27.05° and 27.83°. These peaks are present at a considerably lower fraction of the intensity of the most intense peak of Polymorph 2.

[0040] Polymorph 1 may be converted into a third crystalline polymorph (Polymorph 3) characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 18.57° 2θ as measured using Cu Kα radiation. In particular, Polymorph 1 may be converted to Polymorph 3 under thermal cycling conditions by transitioning between a temperature of 5°C and 50°C. Polymorph 3 may form at a phase transition temperature of about 40°C or under and be obtained as an anhydrate. In a non-limiting example, temperature cycling to produce Polymorph 3 may be conducted in an acetonitrile slurry of Polymorph 1. Polymorph 3 may revert to Polymorph 1 upon heating above about 125°C. Polymorph 3 may be converted to Polymorph 2 upon heating above about 40°C and below about 125°C.

[0041] Polymorph 3 exhibits additional x-ray powder diffraction peaks at the following approximate 2θ values, as measured using Cu Kα radiation: 8.88°, 15.76°, 17.78°, 19.44°, 20.17°, and 20.61°. Each of these peaks is present at a significant fraction of the intensity of the most intense peak of Polymorph 3. One or more additional peaks may be observed at the following approximate 2θ values, as measured using Cu Kα radiation: 7.85°, 13.08°, 13.68°, 22.53°, 22.98°, 23.87°, 27.31°, and 27.91°. These peaks are present at a considerably lower fraction of the intensity of the most intense peak of Polymorph 3.

[0042] Polymorph 1 may be converted into a fourth crystalline polymorph (Polymorph 4) characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 17.62° 2θ, as measured using Cu Kα radiation. In particular, Polymorph 1 may be converted to Polymorph 4 by slow evaporatinn of a methanol solution or other solvents in which Polymorph 1 may be dissolved. Polymorph 4 may be obtained as anhydrate. Polymorph 4 may revert to Polymorph 1 upon heating above about 125°C. Polymorph 4 may convert to Polymorph 2 upon heating above about 40°C and below about 125°C. Below about 40°C, Polymorph 4 may convert to Polymorph 3.

[0043] Polymorph 4 exhibits additional x-ray powder diffraction peaks at the following approximate 2θ values, as measured using Cu Kα radiation: 8.80°, 15.73°, 18.31°, 18.49°, 19.17°, 19.46°, 19.99°, 20.35°, and 20.72°. Each of these peaks is present at a significant fraction of the intensity of the most intense peak of Polymorph 4. One or more additional peaks may be observed at the following approximate 2θ values, as measured using Cu Kα radiation: 7.88°, 13.11°, 13.84°, 21.03°, 21.82°, 27.18°, and 27.86°. These peaks are present at a considerably lower fraction of the intensity of the most intense peak of Polymorph 4.

[0044] Polymorph 1 may be converted into a fifth crystalline polymorph (Polymorph 5) characterized by an x-ray powder diffraction pattern having a most-intense peak at approximately 16.91° 2θ, as measured using Cu Kα radiation. In particular, Polymorph 1 may be converted to Polymorph 5 under thermal cycling conditions by transitioning between a temperature of 5°C and 50°C. Polymorph 5 may be obtained as an anhydrate. In anon-limiting example, temperature cycling to produce Polymorph 5 may be conducted in a 1 :3 THF/heptane (vol.:vol.) slurry or an MTBE slurry of Polymorph 1. Polymorph 5 may revert to Polymorph 1 upon heating above about 125°C. Polymorph 5 may convert to Polymorph 2 upon heating above about 40°C and below about 125°C. Below about 40°C, Polymorph 5 may convert to Polymorph 3.

[0045] Polymorph 5 exhibits additional x-ray powder diffraction peaks at the following approximate 2θ values, as measured using Cu Kα radiation: 8.43°, 17.26°, 17.54°, 18.04°, 18.52°, 19.20°, 19.86°, and 20.70°. Each of these peaks is present at a significant fraction of the intensity of the most intense peak of Polymorph 5. One or more additional peaks may be observed at the following approximate 2θ values, as measured using Cu Kα radiation: 13.92°, 15.94°, 24.40°, 25.79°, 26.45°, and 27.16°. These peaks are present at a considerably lower fraction of the intensity of the most intense peak of Polymorph 5.

[0046] It is to be appreciated that the 2θ peak positions described above are approximate and may vary to some degree depending on sample placement, instrument limitations, and other factors. Characterization of a sample as comprising a particular polymorph may be based upon mapping multiple x-ray powder diffraction peaks upon an unknown polymorphic form onto the known peak positions. The peak positions specified herein may vary by about 0.02° or less, or about 0.05° or less in some instances.

[0047] Pharmaceutical compositions may comprise any of Polymorphs 1-5. Suitable pharmaceutical compositions may comprise additional components, as described hereinafter. Particularly suitable pharmaceutical compositions may comprise Polymorph 3 or Polymorph 1, preferably Polymorph 3.

Pharmaceutical Compositions

[0048] In some aspects, the disclosure provides pharmaceutical compositions comprising any compound herein, or a pharmaceutically acceptable form thereof.

[0049] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0050] The polymorphs described herein may be formulated using one or more pharmaceutically acceptable excipients (carriers) known to persons having ordinary skill in the art. The term "pharmaceutically acceptable excipient," as used herein, refers to substances or components that do not cause unacceptable losses of pharmacological activity or unacceptable adverse side effects when administered to a subject. Example “pharmaceutically acceptable excipients” include, but are not limited to, solvents, dispersion media, coatings, antibacterial agents, antifungal agents, isotonic, and absorption delaying agents, provided that any of these agents do not produce significant side effects or are incompatible with the polymorphs. Example excipients are described, for example, in Remington’s Pharmaceutical Sciences (A.R. Gennaro, Ed.), 21st edition, ISBN: 0781746736 (2005) and United States Pharmacopeia (USP 29) and National Formulary (NF 24), United States Pharmacopeial Convention, Inc, Rockville, Maryland, 2005 ("USP/NF"), or a more recent edition, and the components listed in the continuously updated Inactive Ingredient Search online database of the FDA. Other useful components that are not described in the USP/NF may also be used. Such formulations may contain a therapeutically effective amount of one or more polymorphs, optionally as a salt, hydrate, and/or solvate, together with a suitable amount of excipient to provide a form for proper administration to a subject. The pharmaceutical compositions may be present in solid or liquid form.

[0051] Pharmaceutical compositions of the present disclosure may be stable to specified storage conditions. A "stable" composition refers to a composition having sufficient stability to allow storage at a convenient temperature, such as from about 0°C to about 60°C or about - 20°C to about 50°C, for a commercially reasonable period of time, such as at least about one day, at least about one week, at least about one month, at least about three months, at least about six months, at least about one year, or at least about two years.

[0052] Pharmaceutical compositions of the present disclosure may be tailored to suit a desired mode of administration, which may include, but are not limited to, parenteral, pulmonary, oral, topical, transdermal, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, ophthalmic, pulmonary, epidural, buccal, and rectal. The pharmaceutical compositions may also be administered in combination with one or more additional agents or together with other biologically active or biologically inert agents.

[0053] Controlled-release (or sustained-release) pharmaceutical compositions may be formulated to extend the activity of mitofusin activation and reduce dosing frequency. Controlled-release pharmaceutical compositions may also be used to affect the time of onset of action or other characteristics, such as plasma levels of the mitofusin activator, and consequently affect the occurrence of side effects. Controlled-release pharmaceutical compositions may be designed to initially release an amount of one or more mitofusin activators that produces the desired therapeutic effect, and gradually and continually release other amounts of the mitofusin activator to maintain the level of therapeutic effect over an extended period. In order to maintain a near-constant level of mitofusin activator in the body, the mitofusin activator may be released at a rate sufficient to replace the amount being metabolized or excreted from a subject. The controlled-release may be stimulated by various inducers (e.g, change in pH, change in temperature, enzymes, water, or other physiological conditions or molecules).

[0054] Pharmaceutical compositions described herein may also be used in combination with other therapeutic modalities, as described further below. Thus, in addition to the therapies described herein, one may also provide to the subject other therapies known to be efficacious for treatment of a disease, disorder, or condition being targeted by the mitofusin activator or a related disease, disorder, or condition.

Methods of Use

In some aspects, the present disclosure features a method of treating diseases, disorders, or conditions, comprising administering to a subject in need thereof any compound described herein in a pharmaceutical composition.

[0055] In some aspects, the present disclosure features any compound described herein in a pharmaceutical composition for use for treating diseases, disorders, or conditions, comprising administering to a subject in need thereof.

[0056] In some aspects, the present disclosure features use of any compound described herein in a pharmaceutical composition in the manufacture of a medicament for treating diseases, disorders, or conditions, composing administering to a subject in need thereof. [0057] In some aspects, the present disclosure features a method of activating mitofusin in a subject, comprising administering the compound or the pharmaceutical composition of any one of the preceding claims.

[0058] In some aspects, the present disclosure features any compound described herein in a pharmaceutical composition for use in activating mitofusin in a subject.

[0059] In some aspects, the present disclosure features use of the any compound described herein in a pharmaceutical composition in the manufacture of a medicament for activating mitofusin in a subject.

[0060] In some embodiments, a compound described herein, or any pharmaceutically acceptable form thereof such as a pharmaceutically acceptable salt thereof, can be used to treat or prevent a disease, disorder, or condition in a subject.

[0061] In some embodiments, a therapeutically effective amount of the compound or the pharmaceutical composition described herein is administered to the subject.

[0062] In some embodiments, the disease, disorder, or condition is associated with mitochondria.

[0063] In some embodiments, the subject is human.

[0064] Mitofusin activators of the present disclosure may stimulate mitochondrial fusion, increase mitochondrial fitness, and enhance mitochondrial subcellular transport. Accordingly, in another aspect of the present disclosure, any one or a combination of mitofusin activators of the present disclosure or a pharmaceutically acceptable salt thereof may be administered in a therapeutically effective amount to a subject having or suspected of having a mitochondria- associated disease, disorder or condition. The subject may be a human or other mammal having or suspected of having a mitochondna-associated disease, disorder, or condition. Any of the crystalline polymorphs of the present disclosure may be administered to the subj ect in a suitable form. In a particular example, the mitofusin activator is present as a cry stalline polymorph or a dissolved form thereof, particularly Polymorph 3, as specified above.

[0065] The mitochondria-associated disease, disorder, or condition may be a pheripheral nervous system (PNS) or central nervous system (CNS) genetic or non-genetic disorder, physical damage, and/or chemical injury. In some aspects, in the method of treating a disease, disorder or condition for which a mitofusin activator is indicated, the PNS or CNS disorder may be selected from any one or a combination of: a chronic neurodegenerative condition wherein mitochondrial fusion, fitness, or trafficking are impaired; a disease or disorder associated with mitofusin- 1 (MFN1) or mitofusin-2 (MFN2) dysfunction; a disease associated with mitochondrial fragmentation, dysfunction, or dysmotility; a degenerative neuromuscular condition such as Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis (ALS), Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, hereditary motor and sensory neuropathy, autism, autosomal dominant optic atrophy (ADOA), muscular dystrophy, Lou Gehrig's disease, cancer, mitochondrial myopathy, diabetes mellitus and deafness (DAD), Leber's hereditary optic neuropathy (LHON), Leigh syndrome, subacute sclerosing encephalopathy, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), myoneurogenic gastrointestinal encephalopathy (MNGIE), myoclonic epilepsy with ragged red fibers (MERRF), mitochondrial myopathy, encephalomyopathy, lactic acidosis, stroke-like symptoms (MELAS), mtDNA depletion, mitochondrial neurogastrointestinal encephalomyopathy (MNGIE), dysautonomic mitochondrial myopathy, mitochondrial channelopathy, or pyruvate dehydrogenase complex deficiency (PDCD/PDH), diabetic neuropathy, chemotherapy -induced peripheral neuropathy, crush injury, SCI, traumatic brain injury (TBI), stroke, optic nerve injury, and/or related conditions that involve axonal disconnection.

[0066] Other mitochondria-associated diseases, disorders, or conditions that may be treated with the compositions disclosed herein mlcude, but are not limited to, Alzheimer's disease, ALS, Alexander disease, Alpers' disease, Alpers-Huttenlocher syndrome, alpha-methylacyl- CoA racemase deficiency, Andermann syndrome, Arts syndrome, ataxia neuropathy spectrum, ataxia (e.g., with oculomotor apraxia, autosomal dominant cerebellar ataxia, deafness, and narcolepsy), autosomal recessive spastic ataxia of Charlevoix-Saguenay, Batten disease, beta- propeller protein-associated neurodegeneration, cerebro-oculo-facio-skeletal syndrome (COFS), corticobasal degeneration, CLN1 disease, CLN10 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN6 disease, CLN7 disease, CLN8 disease, cognitive dysfunction, congenital insensitivity to pain with anhidrosis, dementia, familial encephalopathy with neuroserpm inclusion bodies, familial British dementia, familial Danish dementia, fatty acid hydroxylase-associated neurodegeneration, Friedreich’s Ataxia, Gerstmann-Straussler- Scheinker Disease, GM2 -gangliosidosis (e.g., AB variant), HMSN type 7 (e.g., with retinitis pigmentosa), Huntington's disease, infantile neuroaxonal dystrophy, infantile-onset ascending hereditary spastic paralysis, infantile-onset spinocerebellar ataxia, juvenile primary lateral sclerosis, Kennedy's disease, Kuru, Leigh's Disease, Marinesco-Sjogren syndrome, mild cognitive impairment (MCI), mitochondrial membrane protein-associated neurodegeneration, motor neuron disease, monomelic amyotrophy, motor neuron diseases (MND), multiple system atrophy, multiple system atrophy with orthostatic hypotension (Shy-Drager Syndrome), multiple sclerosis, multiple system atrophy, neurodegeneration in Down’s syndrome (NDS), neurodegeneration of aging, neurodegeneration with brain iron accumulation, neuromyelitis optica, pantothenate kinase-associated neurodegeneration, opsoclonus myoclonus, prion disease, progressive multifocal leukoencephalopathy, Parkinson's disease, Parkinson’s disease- related disorders, polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy, prion disease, progressive external ophthalmoplegia, riboflavin transporter deficiency neuronopathy, Sandhoff disease, spinal muscular atrophy (SMA), spinocerebellar ataxia (SCA), striatonigral degeneration, transmissible spongiform encephalopathies (prion diseases), and/or Wallerian-like degeneration.

[0067] Still other mitochrondria-associated diseases, disorders, or conditions that may be treated with the compositions disclosed herein include abulia; agraphia; alcoholism; alexia; alien hand syndrome; Allan-Hemdon-Dudley syndrome; alternating hemiplegia of childhood; Alzheimer's disease; amaurosis fugax; amnesia; ALS; aneurysm; angelman syndrome; anosognosia; aphasia; apraxia; arachnoiditis; Amold-Chiari malformation; asomatognosia; Asperger syndrome; ataxia; attention deficit hyperactivity disorder; atr-16 syndrome; auditory processing disorder; autism spectrum; Behcets disease; bipolar disorder; Bell's palsy; brachial plexus injury; brain damage; brain injury; brain tumor; Brody myopathy; Canavan disease; capgras delusion; carpal tunnel syndrome; causalgia; central pain syndrome; central pontine myelinolysis; centronuclear myopathy; cephalic disorder; cerebral aneurysm; cerebral arteriosclerosis; cerebral atrophy; cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL); cerebral dysgenesis-neuropathy-ichthyosis- keratoderma syndrome (CEDNIK syndrome); cerebral gigantism; cerebral palsy; cerebral vasculitis; cervical spinal stenosis; Charcot-Marie-Tooth disease; chiari malformation; chorea; chronic fatigue syndrome; chronic inflammatory demyelinating polyneuropathy (CIDP); chronic pain; Cockayne syndrome; Coffm-Lowry syndrome; coma; complex regional pain syndrome; compression neuropathy; congenital facial diplegia; corticobasal degeneration; cranial arteritis; craniosynostosis; Creutzfeldt-Jakob disease; cumulative trauma disorders; Cushing's syndrome; cyclothymic disorder; cyclic vomiting syndrome (CVS); cytomegalic inclusion body disease (CIBD); cytomegalovirus infection; Dandy-Walker syndrome; Dawson disease; de Morsier's syndrome; Dejerine-Klumpke palsy; Dejerine-Sottas disease; delayed sleep phase syndrome; dementia; dermatomyositis; developmental coordination disorder; diabetic neuropathy; diffuse sclerosis; diplopia; disorders of consciousness; down syndrome; Dravet syndrome; Duchenne muscular dystrophy; dysarthria; dysautonomia; dyscalculia; dysgraphia; dyskinesia; dyslexia; dystonia; empty sella syndrome; encephalitis; encephalocele; encephalotrigeminal angiomatosis; encopresis; enuresis; epilepsy; epilepsy-intellectual disability in females; Erb's palsy; erythromelalgia; essential tremor; exploding head syndrome; Fabry's disease; Fahr's syndrome; fainting; familial spastic paralysis; febrile seizures; Fisher syndrome; Friedreich's ataxia; fibromyalgia; Foville's syndrome; fetal alcohol syndrome; fragile x syndrome; fragile x-associated tremor/ataxia syndrome (FXTAS); Gaucher's disease; generalized epilepsy with febrile seizures plus; Gerstmann's syndrome; giant cell arteritis; giant cell inclusion disease; globoid cell leukodystrophy; gray matter heterotopia; Guillain-Barre syndrome; generalized anxiety disorder; HTLV-1 associated myelopathy; Hallervorden-Spatz syndrome; head injury; headache; hemifacial spasm; hereditary spastic paraplegia; heredopathia atactica polyneuritiformis; herpes zoster oticus; herpes zoster; Hirayama syndrome; Hirschsprung's disease; Holmes-Adie syndrome; holoprosencephaly; Huntington's disease; hydranencephaly; hydrocephalus; hypercortisolism; hypoxia; immune-mediated encephalomyelitis; inclusion body myositis; incontinentia pigmenti; infantile refsum disease; infantile spasms; inflammatory myopathy; intracranial cyst; intracranial hypertension; isodicentnc 15; Joubert syndrome; Karak syndrome; Keams-Sayre syndrome; Kinsboume syndrome; Kleine-Levm syndrome; Klippel Feil syndrome; Krabbe disease; Kufor-Rakeb syndrome; Lafora disease; Lambert-Eaton myasthenic syndrome; Landau-Kleffner syndrome; lateral medullary (Wallenberg) syndrome; learning disabilities; Leigh's disease; Lennox- Gastaut syndrome; Lesch-Nyhan syndrome; leukodystrophy; leukoencephalopathy with vanishing white matter; Lewy body dementia; lissencephaly; locked-in syndrome; Lou Gehrig's disease (amyotrophic lateral sclerosis (ALS)); lumbar disc disease; lumbar spinal stenosis; lyme disease - neurological sequelae; Machado-Joseph disease (spinocerebellar ataxia type 3); macrencephaly; macropsia; mal de debarquement; megalencephalic leukoencephalopathy with subcortical cysts; megalencephaly; Melkersson-Rosenthal syndrome; Menieres disease; meningitis; Menkes disease; metachromatic leukodystrophy; microcephaly; micropsia; migraine; Miller Fisher syndrome; mini-stroke (transient ischemic attack); misophonia; mitochondrial myopathy; mobius syndrome; monomelic amyotrophy; Morvan syndrome; motor neurone disease - see ALS; motor skills disorder; moyamoya disease; mucopolysaccharidoses; multi-infarct dementia; multifocal motor neuropathy; multiple sclerosis; multiple system atrophy; muscular dystrophy; myalgic encephalomyelitis; myasthenia gravis; myelinoclastic diffuse sclerosis; myoclonic encephalopathy of infants; myoclonus; myopathy; myotubular myopathy; myotonia congenita; narcolepsy; neuro- Behcet's disease; neurofibromatosis; neuroleptic malignant syndrome; neurological manifestations of aids; neurological sequelae of lupus; neuromyotonia; neuronal ceroid lipofuscinosis; neuronal migration disorders; neuropathy; neurosis; Niemann-Pick disease; non-24-hour sleep-wake disorder; nonverbal learning disorder; O'Sullivan-McLeod syndrome; occipital neuralgia; occult spinal dysraphism sequence; Ohtahara syndrome; olivopontocerebellar atrophy; opsoclonus myoclonus syndrome; optic neuritis; orthostatic hypotension; otosclerosis; overuse syndrome; palinopsia; paresthesia; Parkinson's disease; paramyotonia congenita; paraneoplastic diseases; paroxysmal attacks; Parry-Romberg syndrome; pediatric autoimmune neuropsychiatric disorders associated with streptococcoal infections (PANDAS); Pelizaeus-Merzbacher disease; periodic paralyses; peripheral neuropathy; pervasive developmental disorders; phantom limb/phantom pain; photic sneeze reflex; phytanic acid storage disease; Pick's disease; pinched nerve; pituitary tumors; pmg; polyneuropathy; polio; polymicrogyria; polymyositis; porencephaly; post-polio syndrome; postherpetic neuralgia (phn); postural hypotension; Prader-Willi syndrome; primary lateral sclerosis; prion diseases; progressive hemifacial atrophy; progressive multifocal leukoencephalopathy; progressive supranuclear palsy; prosopagnosia; pseudotumor cerebri; quadrantanopia; quadriplegia; rabies; radiculopathy; Ramsay Hunt syndrome type 1; Ramsay Hunt syndrome type 2; Ramsay Hunt syndrome type 3 - see Ramsay-Hunt syndrome; Rasmussen encephalitis; reflex neurovascular dystrophy; refsum disease; REM sleep behavior disorder; repetitive stress injury; restless legs syndrome; retrovirus-associated myelopathy; Rett syndrome; Reye's syndrome; rhythmic movement disorder; Romberg syndrome; Saint Vitus’ dance; Sandhoff disease; Schilder's disease (two distinct conditions); schizencephaly; sensory processing disorder; septo-optic dysplasia; shaken baby syndrome; shingles; Shy- Drager syndrome; Sjogren's syndrome; sleep apnea; sleeping sickness; snatiation; Sotos syndrome; spasticity; spina bifida; spinal cord injury; spinal cord tumors; spinal muscular atrophy; spinal and bulbar muscular atrophy; spinocerebellar ataxia; split-brain; Steele- Richardson-Olszewski syndrome; stiff-person syndrome; stroke; Sturge-Weber syndrome; stuttering; subacute sclerosing panencephahtis; subcortical arteriosclerotic encephalopathy; superficial siderosis; Sydenham's chorea; syncope; synesthesia; syringomyelia; tarsal tunnel syndrome; tardive dyskinesia; tardive dysphrenia; Tarlov cyst; Tay-Sachs disease; temporal arteritis; temporal lobe epilepsy; tetanus; tethered spinal cord syndrome; Thomsen disease; thoracic outlet syndrome; tic douloureux; Todd's Paralysis; Tourette syndrome; toxic encephalopathy; transient ischemic attack; transmissible spongiform encephalopathies; transverse myelitis; traumatic brain injury; tremor; trichotillomania; trigeminal neuralgia; tropical spastic paraparesis; trypanosomiasis; tuberous sclerosis; 22ql3 deletion syndrome; Unverricht-Lundborg disease; vestibular schwannoma (acoustic neuroma); Von Hippel- Lindau disease (VHL); viliuisk encephalomyelitis (VE); Wallenberg's syndrome; west syndrome; whiplash; Williams syndrome; Wilson's disease; y-linked hearing impairment; and/or Zellweger syndrome.

[0068] Each of the states, diseases, disorders, and conditions, described herein, as well as others, can benefit from compositions and methods described herein. Generally, treating a state, disease, disorder, or condition includes preventing or delaying the appearance of clinical symptoms in a mammal that may be afflicted with or predisposed to the state, disease, disorder, or condition but does not yet experience or display clinical or subclinical symptoms thereof. Treating can also include inhibiting the state, disease, disorder, or condition (e.g., arresting or reducing the development of the disease or at least one clinical or subclinical symptom thereof). Furthermore, treating can include relieving the disease (e.g., causing regression of the state, disease, disorder, or condition or at least one of its clinical or subclinical symptoms). A benefit to a subject to be treated can be either statistically significant or at least perceptible to the subject or to a physician.

[0069] A mitochondria-associated disease, disorder, or condition may be a disease primarily caused by or secondarily associated with mitochondrial dysfunction, fragmentation, or loss-of- fusion, or associated with dysfunction in MFN1 or MFN2 catalytic activity or conformational unfolding. Mitochondrial dysfunction may be caused by genetic mutations of mitofusins or other (nuclear or mitochondrial encoded) genes, or may be caused by physical, chemical, or environmental injury to the CNS or PNS.

[0070] In a particular example, cancer chemotherapy-induced sensory and motor neuropathies may be prevented or treated with the compositions of the present disclosure. Chemotherapy- induced peripheral neuropathy is one of the most common complications of cancer chemotherapy, affecting 20% of all patients and almost 100% of patients receiving high doses of chemotherapeutic agents. Dose-dependent neurotoxicity of motor and sensory neurons can lead to chronic pain, hypersensitivity to hot, cold, and mechanical stimuli, and/or impaired neuromuscular control. The most common chemotherapeutic agents linked to CIPN are platinum, vinca alkaloids, taxanes, epothilones, and the targeted proteasome inhibitor, bortezomib.

[0071] CIPN most commonly affects peripheral sensory neurons whose cell bodies are located in dorsal root ganglia lacking the blood-brain barrier that protects other components of the central and peripheral nervous system. Unprotected dorsal root ganglion neurons are more sensitive to neuronal hyperexcitability and innate immune system activation evoked by circulating cytotoxic chemotherapeutic agents. CIPN affects quality of life, and is potentially disabling, because it provokes chronic neuropathic pain that, like other causes of neuralgia (e.g., post herpetic neuralgia, diabetic mononeuropathy), is refractory to analgesic therapy. Motor nerve involvement commonly manifests as loss of fine motor function with deterioration in hand writing, difficulty in buttoning clothes or sewing, and sometimes upper and lower extremity weakness or loss of endurance. CIPN typically manifests within weeks of chemotherapy and in many cases improves after chemotherapy treatment ends, although residual pain, sensory, or motor defects are observed in one-third to one-half of affected patients. Unfortunately, CIPN-limited chemotherapy dosing can lead to delays, reduction, or interruption of cancer treatment, thus shortening survival.

[0072] Mitochondrial dysfunction and oxidative stress are implicated in CIPN because of observed ultrastructural morphological abnormalities, impaired mitochondria DNA transcription and replication, induction of mitochondrial apoptosis pathways, and reduction of experimental CIPN signs by anticipatory mitochondrial protection. Mitofusin activators may enhance overall mitochondrial function in damaged neurons, increase mitochondrial transport to areas of neuronal damage, and accelerate in vitro neuron repair/regeneration after chemotherapy-induced damage. For this reason, it is believed that mitofusin activators may reduce neuronal injury conferred by chemotherapeutic agents in CIPN and accelerate regeneration/repair of nerves damaged by chemotherapeutic anticancer agents. As such, the present disclosure provides for compositions and methods to treat cancer chemotherapy induced nerve injury and neuropathy.

[0073] In another example, injury in the CNS or PNS (e.g., trauma to the CNS or PNS, crush injury, SCI, TBI, stroke, optic nerve injury, or related conditions that involve axonal disconnection) may be treated with the compositions of the present disclosure. The CNS includes the brain and the spinal cord and the PNS is composed of cranial, spinal, and autonomic nerves that connect to the CNS.

[0074] Damage to the nervous system caused by mechanical, thermal, chemical, or ischemic factors may impair various nervous system functions such as memory, cognition, language, and voluntary movement. Most often, this is through accidental crush or transection of nerve tracts, or as an unintended consequence of medical interventions, that interrupt normal communications between nerve cell bodies and their targets. Other types of injuries may include disruption of the interrelations between neurons and their supporting cells or the destruction of the blood-brain barrier.

[0075] Mitofusin activators may rapidly reverse mitochondrial dysmotility in neurons from mice or patients with various genetic or chemotherapeutic neurodegenerative diseases, in axons injured by chemotherapeutic agents, and in axons severed by physical injury. For this reason, mitofusin activators may enhance regeneration/repair of physically damaged nerves, as in vehicular and sports injuries, penetration trauma from military or criminal actions, and iatrogenic injury during invasive medical procedures. As such, the present disclosure provides for compositions and methods to treat physical nerve injury.

[0076] Mitochondrial motility is also implicated in neuropathy and traumatic crush or severance nerve injuries. After nerve laceration or crush injury, nerves will either regenerate and restore neuromuscular function or fail to regenerate such that neuromuscular function in permanently impaired. Mitofusin activators may increase mitochondrial trafficking, thereby enabling a nerve to regenerate after traumatic injuries.

[0077] The amount of a mitofusin activator and excipient to produce a pharmaceutical composition in a given dosage form may vary depending upon the subject being treated, the condition being treated, and the particular mode of administration. It will be appreciated that the unit content of mitofusin activator contained in an individual dose of a given dosage form need not in itself constitute a therapeutically effective amount, as the necessary therapeutically effective amount could be reached by administration of a number of individual doses, or the therapeutic effect may be cumulative over time.

[0078] Dosing of the mitofusin activators of the present disclosure may occur as a single event or over a time course of treatment. For example, a mitofusin activator may be administered daily, weekly, bi-weekly, or monthly. For treatment of acute conditions, the time course of treatment may be at least several days, with dosing taking place at least once a day or continuously. Certain conditions could extend treatment from several days to several weeks. For example, treatment could extend over one week, two weeks, or three weeks. For chronic conditions, treatment could extend from several weeks to several months or even years.

[0079] Toxicity and therapeutic efficacy of the compositions described herein may be determined by standard pharmaceutical procedures in cell cultures or experimental animals for determining the LDso (the dose lethal to 50% of the population) and the ED50, (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index that may be expressed as the ratio LD50/ED50, where larger therapeutic indices are generally understood in the art to be optimal. For the polymorphic forms of the mitofusin activators disclosed herein, the therapeutic index may be about 30 or greater. Definitions

[0080] The terms “treat” or “treatment”, unless otherwise indicated by context, refer to any administration of a therapeutic molecule (e.g., any compound described herein) that partially or completely alleviates, ameliorates, relieves, inhibits, reduces severity of and/or reduces incidence of one or more symptoms or features of a particular disease, disorder, and/or condition.

[0081] As used herein, the term “preventing,” “prevent,” or “protecting against” describes delaying onset or slowing progression of a disease, condition or disorder.

[0082] As used herein, the term “subject” includes human and non-human animals, as well as cell lines, cell cultures, tissues, and organs. In some embodiments, the subject is a mammal. The mammal can be e.g. , a human or appropriate non-human mammal, such as primate, mouse, rat, dog, cat, cow, horse, goat, camel, sheep or a pig. The subject can also be a bird or fowl. In some embodiments, the subject is a human.

[0083] As used herein, the term “subject in need thereof’ refers to a subject having a disease or having an increased risk of developing the disease. A subject in need thereof can be one who has been previously diagnosed or identified as having a disease or disorder disclosed herein. A subject in need thereof can also be one who is suffering from a disease or disorder disclosed herein. Alternatively, a subject in need thereof can be one who has an increased risk of developing such disease or disorder relative to the population at large (i.e., a subject who is predisposed to developing such disorder relative to the population at large). A subject in need thereof can have a refractory or resistant a disease or disorder disclosed herein (i.e., a disease or disorder disclosed herein that does not respond or has not yet responded to treatment). The subject may be resistant at start of treatment or may become resistant dunng treatment. In some embodiments, the subject in need thereof received and failed all known effective therapies for a disease or disorder disclosed herein. In some embodiments, the subject in need thereof received at least one prior therapy.

[0084] The term “therapeutically effective amount” or “effective amount” refers to an amount of a conjugate effective to treat or prevent a disease or disorder in a subject (e.g., as described herein).

[0085] As used herein, the term “pharmaceutical composition” refers to a composition in which an active agent is formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, a pharmaceutical composition may be specially formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream, or foam; sublingually; ocularly; transdermally; or nasally, pulmonary, and to other mucosal surfaces.

[0086] As used herein, the term “administration” typically refers to the administration of a composition to a subject or system to achieve delivery of an agent that is, or is included in, the composition. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, etc. In some embodiments, administration is parenteral (e.g., intravenous administration). In some embodiments, intravenous administration is intravenous infusion. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e. g. intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc.

[0087] As used herein, the term “pharmaceutically acceptable salt” refers to organic or inorganic salts of a polymorph of the present disclosure that have specified toxicity and/or biodistribution properties. Suitable salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p- toluenesulfonate, and/or pamoate (i.e., l,l'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. The pharmaceutically acceptable salt may balance charge on the parent polymorph by being present as a counterion. More than one counterion may be present. When multiple counterions are present, the polymorphs may be present as a mixed pharmaceutically acceptable salt. [0088] Pharmaceutically acceptable salts, solvates, and/or hydrates of any of Polymorphs 1-5 may be present in the pharmaceutical compositions of the present disclosure. As used herein, the term “pharmaceutically acceptable solvate” refers to an association between one or more solvent molecules and a polymorph of the present disclosure or a salt thereof, wherein the solvate has specified toxicity and/or biodistribution properties. Examples of solvents that may form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and/or ethanolamine. As used herein, the term “pharmaceutically acceptable hydrate” refers to a polymorph of the present disclosure or a salt thereof that further includes a stoichiometric or non-stoichiometric amount of water bound by non-covalent mtermolecular forces, wherein the hydrate has specified toxicity and/or biodistribution properties. Preferably, the polymorphs of the present disclosure are substantially non-solvated and substantially non-hydrated when present in the pharmaceutical compositions described herein.

[0089] To facilitate a better understanding of the present disclosure, the following examples of preferred or representative embodiments are given. In no way should the following examples be read to limit, or to define, the scope of the invention.

EXAMPLES

[0090] X-ray powder diffraction patterns were obtained on a Panalytical XTert Powder system on a Si zero-background sample holder. The 2θ position was calculated against a Pananalytical Si reference standard disc. Other experimental parameters are set forth in Table 1 below.

Table 1

[0091] Thermogravimetric analysis (TGA) data was collected using a TA Discovery 550 TGA from TA Instrument. Differential scanning calorimetry (DSC) was performed using a TA 2500 DSC from TA Instrument. DSC was calibrated with an indium reference standard and TGA was calibrated using nickel reference standard. Detailed parameters used are listed in Table 2.

Table 2

[0092] Polarized light microscopy images were obtained at room temperature using a Nikon

DS-Fi2 upright microscope.

[0093] Synthesis and Characterization of N-(trans-4-hydroxycyclohexyl)-5- phenylpentanamide (Polymorph 1). This mitofusin activator was prepared as described in U.S. Patent Application Publication 2020/0345668, which is incorporated herein by reference. FIG. 1 shows an illustrative x-ray powder diffraction pattern of as-produced N-(trans-4- hydroxycyclohexyl)-5-phenylpentanamide. The as-produced polymorph form of N-(trans-4- hydroxycyclohexyl)-5-phenylpentanamide is designated as Polymorph 1 herein. As shown, this polymorph may be characterized by its most-intense peak at approximately 18.43° 2θ. Table 3 shows additional peak positions in the x-ray powder diffraction pattern of Polymorph 1 and their relative intensity compared to the most intense peak.

Table 3

[0094] FIG. 2 shows an illustrative polarized light microscopy image of as-produced N -(trans- 4-hydroxycyclohexyl)-5-phenylpentanamide (Polymorph 1). As shown. Polymorph 1 exhibited a rod-like morphology. TGA and DSC (not shown) showed a single thermal event at approximately 132°C, indicative of melting, and afforded negligible weight loss, leading to an anhydrate designation for this polymorphic form. No change was observed in the x-ray powder diffraction pattern upon heating to 125°C. Polymorph 1 was also obtained by layering an anti-solvent onto a solution of N -(trans--4-hydroxycyclohexyl)-5-phenylpentanamide and allowing crystallization to take place.

[0095] Conversion of Polymorph 1 into Polymorph 2. As-produced Polymorph 1 of N-(trans--4-hydroxycyclohexyl)-5-phenylpentanamide was converted into a polymorph having a most-intense x-ray powder diffraction peak at approximately 17.44° 2θ by temperature cycling a 1:2 ethanol/heptane (vol.wol.) slurry, as well as under several other temperature cycling conditions (alternating 5°C and 50°C cycles). FIG. 3 shows an illustrative x-ray powder diffraction pattern of Polymorph 2 of N-(trans--4-hydroxycyclohexyl)-5-phenylpentanamide. characterized by its most-intense peak at approximately 17.44° 2θ. Table 4 shows additional peak positions in the x-ray powder diffraction pattern of Polymorph 2 and their relative intensity compared to the most intense peak.

Table 4 [0096] FIG. 4 shows an illustrative polarized light microscopy image of Polymorph 2 of N-(trans--4-hydroxycyclohexyl)-5-phenylpentanamide. As shown, Polymorph 2 had a small, needle-like morphology. FIG. 5 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 2 of N-(trans-4- hydroxycyclohexyl)-5-phenylpentanamide. The small endotherm at 123.4°C and exotherm at 128.8°C are believed to result from concurrent melting and recrystallization of Polymorph 2 to reform Polymorph 1. The endotherm at 133.1°C is believed to result from melting of Polymorph 1. Otherwise, based upon negligible weight loss, the thermal data is consistent with designation of Polymorph 2 as an anhydrate. Upon heating Polymorph 2 to 125°C, Polymorph 1 reformed, as evidenced by reversion of the x-ray powder diffraction pattern to that shown in FIG. 1.

[0097] Conversion of Polymorph 1 into Polymorph 3. As-produced Polymorph 1 of N-(trans--4-hydroxycyclohexyl)-5-phenylpentanamide was converted into a polymorph having a most-intense x-ray powder diffraction peak at approximately 18.57° 2θ by temperature cycling an acetonitrile slurry. FIG. 6 shows an illustrative x-ray powder diffraction pattern of Polymorph 3 of N -(trans--4-hydroxycyclohexyl)-5-phenylpentanamide, characterized by its most-intense peak at approximately 18.57° 2θ. Table 5 shows additional peak positions in the x-ray powder diffraction pattern of Polymorph 3 and their relative intensity compared to the most intense peak.

Table 5 [0098] FIG. 7 shows an illustrative polarized light microscopy image of Polymorph 3 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide. As shown, Polymorph 3 consisted of small agglomerated particles. FIG. 8 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 3 of N-(trans-4- hydroxycyclohexyl)-5-phenylpentanamide. The endothermic events in the TGA/DSC plot are believed to result from reformation of Polymorph 1 at elevated temperatures. Otherwise, based upon negligible weight loss, the thermal data is consistent with designation of Polymorph 3 as an anhydrate. Upon heating Polymorph 3 to 125°C, Polymorph 1 reformed, as evidenced by reversion of the x-ray powder diffraction pattern to that shown in FIG. 1.

[0099] Conversion of Polymorph 1 into Polymorph 4. As-produced Polymorph 1 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide was converted into a polymorph having a most-intense x-ray powder diffraction peak at approximately 17.62° 2θ by slow evaporation of a methanol solution, as well as under several other conditions featuring recrystallization from an organic solvent. FIG. 9 shows an illustrative x-ray powder diffraction pattern of Polymorph 4 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide, characterized by its most-intense peak at approximately 17.62° 2θ. Table 6 shows additional peak positions in the x-ray powder diffraction pattern of Polymorph 4 and their relative intensity compared to the most intense peak.

Table 6 [00100] FIG. 10 shows an illustrative polarized light microscopy image of Polymorph 4 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide. As shown, Polymorph 4 consisted of small needle-like particles. FIG. 11 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 4 of N-(trans-4- hydroxycyclohexyl)-5-phenylpentanamide. The endothermic events in the TGA/DSC plot are believed to result from reformation of Polymorph 1 at elevated temperature. Otherwise, based upon negligible weight loss, the thermal data is consistent with designation of Polymorph 4 as an anhydrate. Upon heating Polymorph 4 to 125°C, Polymorph 1 reformed, as evidenced by reversion of the x-ray powder diffraction pattern to that shown in FIG. 1.

[00101] Conversion of Polymorph 1 into Polymorph 5. As-produced Polymorph 1 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide was converted into a polymorph having a most-intense x-ray powder diffraction peak at approximately 16.91° 2Q by temperature cycling a 1:3 THF/heptane slurry (vol.wol.) or aMTBE slurry, FIG. 12 shows an illustrative x-ray powder diffraction pattern of Polymorph 5 of N-(trans-4-hydroxycyclohexyl)-5- phenylpentanamide, characterized by its most-intense peak at approximately 16.91° 2Q. Table 7 shows additional peak positions in the x-ray powder diffraction pattern of Polymorph 5 and their relative intensity compared to the most intense peak.

Table 7

[00102] FIG. 13 shows an illustrative polarized light microscopy image of Polymorph 5 of N-(trans-4-hydroxycyclohexyl)-5-phenylpentanamide. As shown, Polymorph 5 consisted of small needle-like crystals. FIG. 14 shows an illustrative thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) plot of Polymorph 5 of N- N-(trans-4 hydroxycyclohexyl)-5-phenylpentanamide. The endothermic events in the TGA/DSC plot are believed to result from reformation of Polymorph 1 at elevated temperature. Otherwise, based upon negligible weight loss, the thermal data is consistent with designation of Polymorph 5 as an anhydrate.

[00103] Competitive Slurry Experiments. A saturated stock solution of Polymorph 1 in methyl isobutyl ketone was prepared, and separate suspensions containing an equivalent amount of Polymorphs 1-5 were prepared from the Polymorph 1 saturated stock solution. In each case, the resulting slurry was maintained at room temperature or 60°C for one or three days, and the x-ray powder diffraction patterns of the residue were checked at these times. Polymorph 3 formed as the most stable phase at room temperature, and Polymorph 2 formed as the most stable phase at 60°C.

[00104] A saturated stock solution of Polymorph 1 in methyl isobutyl ketone was prepared, and separate suspensions containing an equivalent amount of Polymorph 2 or Polymorph 3 were prepared from the Polymorph 1 saturated stock solution. The suspensions were maintained at 30°C, 40°C or 50°C for one or three days, and the x-ray powder diffraction patterns of the residue were checked at these times. Polymorph 3 formed at 30°C and 40°C from each suspension. Polymorph 2 formed at 50°C from each suspension. Collectively, the data suggest a phase transition temperature between 40°C and 50°C. Below 40°C, Polymorph 3 is the most stable polymorph, and Polymorph 2 is the most stable polymorph between about 40°C and about 120°C. Above about 120°C, Polymorph 1 is the predominant polymorph. [00105] Solubility Testing. Solubility testing was conducted by placing 2 mg of a polymorph sample into a 3 mL glass vial, and solvent was added stepwise (50 □L, 50 □L, 200 □ L, 700 □L) until the solids dissolved or a total volume of 1 mL was reached. Solubility was estimated visually based upon the amounts of polymorph sample and solvent added. Solubility was determined at room temperature. Table 8 summarizes the approximate solubility of Polymorph 1 in various solvents at room temperature.

Table 8

[00106] Because Polymorph 3 was the most low-temperature stable polymorph, more detailed solubility testing data was collected for this polymorph in a range of vehicles potentially applicable for biological testing. Table 9 summarizes the approximate solubility of Polymorph 3 in various vehicles at room temperature. The approximate solubility was determined in a similar manner to that described above for Polymorph 1.

Table 9

[00107] Aqueous solubility of Polymorph 3 was determined by adding excess Polymorph 3 to a glass HPLC vial and adding a known quantity of water to make a slurry. After stirring at room temperature for 24 hours, the remaining solid was removed through centrifugation. The supernatant was passed through a 0.2 □m filter and diluted for HPLC analysis. The saturation concentration of Polymorph 3 in water was found to be 0.155 mg/mL. No phase conversion was found for the residue solid.

[00108] More detailed solubility testing for Polymorph 3 was collected by placing a known quantity of Polymorph 3 in a glass HPLC vial, followed by a set quantity of vehicle. If full dissolution was initially achieved, additional Polymorph 3 was added until a suspension was obtained. Stirring was then conducted at room temperature for 24 hours. Solids were removed by centrifugation, and the supernatant was passed through a 0.2 □m filter and diluted for HPLC analysis to determine concentration. No phase conversion was found for the residue solid. Table 10 summarizes the detailed solubility of Polymorph 3 in various vehicles at room temperature.

Table 10 Entries marked with an asterisk were tested for compatibility with SGF (NaCl), FaSSIF (phosphate), and FeSSIF (acetate) biological media. All except 20% TPGS in Capmul were compatible with these biological media when mixed in a 1:5 vol.wol. ratio. Table 11 summarizes the saturation solubility of Polymorph 3 when mixed with 1:5 vol.wol. vehicle:biologically relevant medium.

Table 11

EQUIVALENTS

[00109] The details of one or more embodiments of the disclosure are set forth in the accompanying description above. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Other features, objects, and advantages of the disclosure will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms include plural referents unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited in this specification are incorporated by reference.

[00110] The foregoing description has been presented only for the purposes of illustration and is not intended to limit the disclosure to the precise form disclosed, but by the claims appended hereto.

[00111] Unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the embodiments of the present invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claim, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[00112] One or more illustrative embodiments incorporating various features are presented herein. Not all features of a physical implementation are described or shown in this application for the sake of clarity. It is understood that in the development of a physical embodiment incorporating the embodiments of the present invention, numerous implementation-specific decisions must be made to achieve the developer's goals, such as compliance with system-related, business-related, government-related and other constraints, which vary by implementation and from time to time. While a developer's efforts might be time-consuming, such efforts would be, nevertheless, a routine undertaking for those of ordinary skill in the art and having benefit of this disclosure.

[00113] While various systems, tools and methods are described herein in terms of “comprising” various components or steps, the systems, tools and methods can also “consist essentially of’ or “consist of’ the various components and steps.

[00114] As used herein, the phrase “at least one of’ preceding a series of items, with the terms “and” or “or” to separate any of the items, modifies the list as a whole, rather than each member of the list (i.e., each item). The phrase “at least one of’ allows a meaning that includes at least one of any one of the items, and/or at least one of any combination of the items, and/or at least one of each of the items. By way of example, the phrases “at least one of A, B, and C” or “at least one of A, B, or C” each refer to only A, only B, or only C; any combination of A, B, and C; and/or at least one of each of A, B, and C.

[00115] Therefore, the disclosed systems, tools and methods are well adapted to attain the ends and advantages mentioned as well as those that are inherent therein. The particular embodiments disclosed above are illustrative only, as the teachings of the present disclosure may be modified and practiced in different but equivalent manners apparent to those skilled in the art having the benefit of the teachings herein. Furthermore, no limitations are intended to the details of construction or design herein shown, other than as described in the claims below. It is therefore evident that the particular illustrative embodiments disclosed above may be altered, combined, or modified and all such variations are considered within the scope of the present disclosure. The systems, tools and methods illustratively disclosed herein may suitably be practiced in the absence of any element that is not specifically disclosed herein and/or any optional element disclosed herein. While systems, tools and methods are described in terms of “comprising,” “containing,” or “including” various components or steps, the systems, tools and methods can also “consist essentially of’ or “consist of’ the various components and steps. All numbers and ranges disclosed above may vary by some amount. Whenever a numerical range with a lower limit and an upper limit is disclosed, any number and any included range falling within the range is specifically disclosed. In particular, every range of values (of the form, “from about a to about b,” or, equivalently , “from approximately a to b,” or, equivalently, “from approximately a-b”) disclosed herein is to be understood to set forth every number and range encompassed within the broader range of values. Also, the terms in the claims have their plain, ordinary meaning unless otherwise explicitly and clearly defined by the patentee. Moreover, the indefinite articles “a” or “an,” as used in the claims, are defined herein to mean one or more than one of the elements that it introduces. If there is any conflict in the usages of a word or term in this specification and one or more patent or other documents that may be incorporated herein by reference, the definitions that are consistent with this specification should be adopted.