Novel polymorphs and uses thereof

Abstract

In one embodiment, the present application discloses a compound that is a selective neuroactive agent for the treatment of diseases of the central nervous system (CNS). In one aspect, the neuroactive agent is a composition comprising polymorph SP. The present invention relates to Cu II -diacetyl-bis(N 4 -methyl-thiosemicarbazone)(diacetyl-bis(N 4 -Methyl-thiosemicarbazonato)-Cu II , Cu II (atsm), copper ATSM, and CuATSM, the latter term being used herein), as a pharmaceutical, particularly for the treatment of conditions that can be prevented, alleviated, or ameliorated by delivery of the metal.

Description

[Technical Field]

[0001] Background of the application The present invention is II -diacetyl-bis(N 4 -methyl-thiosemicarbazone)(diacetyl-bis(N 4 -Methyl-thiosemicarbazonato)-Cu II , Cu II This invention relates to the use of copper ATSM (also known as copper ATSM, copper ATSM, and copper ATSM, the latter term being used herein) as a pharmaceutical, particularly for the treatment of conditions that can be prevented, alleviated, or ameliorated by delivery of a metal. There are several clinical conditions that are caused by or associated with abnormal levels of metals (typically low metal levels). Conditions of this type include cancer and conditions characterized by or associated with oxidative damage, more specifically neurodegenerative conditions or diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, hypoxia, and prion disease (PrD). [Background technology]

[0002] Bioavailable metals have a profound impact on the function of biological systems. Metals are known to play a major role in enzyme systems and signaling mechanisms within biological systems. For example, Zn plays a key role in the formation of β-amyloid plaques in Alzheimer's disease; the influence of the (Cu, Zn) superoxide dismutase enzyme in mediating reactive oxygen species damage associated with amyotrophic lateral sclerosis; the involvement of the heme enzymes NO synthase and guanylyl cyclase in the production and sensing of nitric oxide (NO), respectively; and the discovery of "zinc finger" motifs in breast and ovarian cancer susceptibility genes (e.g., BRCA1). Furthermore, evidence has demonstrated the propensity of abnormal proteins to misfold in the presence of certain concentrations of metal ions.

[0003] Several cardiovascular conditions have been identified that result in oxidative stress (OS). Other conditions associated with OS include cancer, cataracts, neurodegenerative disorders (such as Alzheimer's disease), and heart disease. There is also evidence that OS plays a prominent role in neuromuscular disorders, including amyotrophic lateral sclerosis (ALS), mitochondrial / metabolic disorders, Friedreich's ataxia, Parkinson's disease, and dementia with Lewy bodies. Common features of these diseases include the deposition of misfolded proteins and substantial cellular injury as a result of OS. Data suggest that OS is a major contributor to physical injury in a wide range of pathologies, including amyloidogenic neurological disorders (Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), prion diseases (including Creutzfeldt-Jakob disease (CJD)), transmissible spongioform encephalopathies (TSEs), cataracts, mitochondrial disorders, Menkes disease, Parkinson's disease (PD), and Huntington's disease (HD)). The effects of OS are not limited to any one part of the human body; examples of its negative effects can be found in almost every organ. For example, the human brain is an organ with high concentrations of metal ions, and recent evidence suggests that disruption of metal homeostasis plays an important role in various age-related neurodegenerative diseases.

[0004] Several therapeutic agents have been developed as potential treatments for symptoms caused by or related to OS. However, drugs such as vitamin E and vitamin C have been found to be ineffective because they do not cross the blood-brain barrier and therefore cannot be effectively used to treat neurodegenerative diseases of central origin.

[0005] Copper metal ion deficiency has been reported as a symptom associated with neurodegenerative diseases (e.g., ALS, PD, and dementia with Lewy bodies). One consequence of copper deficiency is that protective enzymes responsible for detoxifying reactive oxygen species (ROS) are insufficiently loaded with copper and therefore do not perform their normal enzymatic function effectively. Insufficient loading of such protective enzymes in the brain, for example, typically leads to increased OS (as seen in AD), which is reflected by increased protein oxidation (e.g., increased carbonylated proteins).

[0006] Thus, there is a need for highly effective agents (such as CuATSM) for the treatment of diseases involving oxidative damage, particularly neurodegenerative disorders of the central nervous system (such as PD, AD, and ALS). Additionally, there is a need for novel agents for the treatment of conditions involving peripheral tissues, gastrointestinal dysfunction (such as constipation), and acute respiratory distress syndrome, ALS, atherosclerotic cardiovascular disease, and multi-organ dysfunction. There is also a need for commercially viable processes for making such novel agents.

[0007] Although CuATSM has been shown to be effective in diseases involving oxidative damage, particularly neurodegenerative disorders of the central nervous system (e.g., PD and ALS), CuATSM itself is substantially insoluble in aqueous media. Therefore, there is a need to provide CuATSM in a form that is more soluble in aqueous media and / or that can be delivered in an orally available formulation. Summary of the Invention [Means for solving the problem]

[0008] Summary of the application There remains a need for new and effective drugs that are selective neuroactive agents for the treatment of diseases of the central nervous system (CNS). Accordingly, a first object of the present invention is to provide stable polymorphs of CuATSM:gluconic acid. A further object of the present invention is to provide a safe and commercially viable process for the preparation of the stable polymorphs.

[0009] Therefore, Cu II -diacetyl-bis(N4 Described herein are stable polymorphs of CuATSM (CuATSM-methyl-thiosemicarbazone) and gluconic acid, having an X-ray powder diffraction ("XRPD") spectrum comprising peaks at 2-theta angles of 7.5°, 9°, and 11°. In some embodiments, the XRPD spectrum comprises additional peaks at 15.5°, 27.5°, 28.5°, and 32°. The polymorph typically has at least five XPRD spectrum peaks selected from the group consisting of 2-theta angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. Alternatively, the polymorph has at least six XPRD spectrum peaks selected from the group consisting of 2-theta angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. The polymorph may have peaks in the XPRD spectrum at approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32° 2-theta angles.

[0010] Further described herein are methods for treating or preventing a condition in a mammal, wherein delivery of copper prevents, alleviates, or ameliorates the condition, comprising administering to the mammal a therapeutically effective amount of a composition comprising CuATSM and a stable polymorph of gluconic acid, wherein the composition has an XRPD spectrum comprising peaks at 2-theta angles of 7.5°, 9°, and 11°; and a pharmaceutically acceptable excipient. The composition may comprise a polymorph having at least five XPRD spectral peaks selected from the group consisting of 2-theta angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. Alternatively, the composition may comprise a polymorph having at least six XPRD spectral peaks selected from the group consisting of 2-theta angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. The composition may include polymorphs having peaks in the XPRD spectrum at 2θ angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°.

[0011] Also described herein is a process for preparing a stable polymorph of CuATSM and gluconic acid, the process comprising: (a) mixing ATSMH2 and copper gluconate (in a first ratio) with a solvent (in a second ratio) to form a slurry; (b) heating said slurry to form a composition comprising polymorph SP; and (c) isolating said polymorph.

[0012] Further described herein is a stable polymorph of CuATSM and gluconic acid, made by a process comprising: (a) combining ATSMH2 and copper gluconate (in a first ratio) with a solvent (in a second ratio) to form a slurry; (b) heating said slurry to form said composition; and (c) isolating said polymorph. [Brief explanation of the drawings]

[0013] [Figure 1] Figure 1 is an X-ray powder diffraction ("XRPD") spectrum for the product made in Example 1. The spectrum for the product is shown at the top of Figure 1 and compared below with the XRPD spectra of CuATSM, copper D-gluconate, D-gluconic acid lactone, and D-gluconic acid, respectively.

[0014] [Figure 2] Figure 2 is an XRPD spectrum for the product made in Example 2. The spectrum for the product is shown at the top of Figure 2 and compared with the XRPD spectra of CuATSM, copper D-gluconate, and D-gluconic acid lactone below, respectively.

[0015] [Figure 3] Figure 3 is an XRPD spectrum for the product made in Example 3. The spectrum for the product is shown at the top of Figure 3 and compared with the XRPD spectra of CuATSM, copper D-gluconate, and D-gluconic acid lactone below, respectively.

[0016] [Figure 4]Figure 4 is an XRPD spectrum for the product made in Example 4. The spectrum for the product is shown at the top of Figure 4 and compared with the XRPD spectra of copper D-gluconate and gluconic acid lactone below, respectively. DETAILED DESCRIPTION OF THE INVENTION

[0017] Detailed description of the application This application discloses stable polymorphs of compositions containing CuATSM and gluconic acid. II -diacetyl-bis(N 4 -methyl-thiosemicarbazone), diacetyl-bis(N 4 -Methyl-thiosemicarbazonato)-Cu II , Cu II (atsm), or copper ATSM, has the following structure: [ka] The stable polymorphs of CuATSM and gluconic acid are also referred to herein as polymorph SP. The stable polymorphs have the distinctive XRPD spectra disclosed herein. In some embodiments, the stable polymorphs have XRPD spectra that include peaks at 2θ angles of 7.5°, 9°, and 11°. In some embodiments, in addition to these peaks, the stable polymorphs have XRPD spectra that further include peaks at 2θ angles of 15.5°, 27.5°, 28.5°, and 32°. The "2θ angle" values ​​provided herein are meant to be a best approximation of the peak values ​​provided at the top of Figures 1, 3, and 4 herein.

[0018] This application discloses the use of polymorph SP to deliver copper metal to biological sites, tissues, or cells in patients with copper depletion. Several important copper-mediated biological processes (such as copper-mediated enzymatic processes) occur intracellularly rather than in the extracellular matrix. Copper is delivered in the form of polymorph SP to ensure that copper acts intracellularly rather than in the extracellular environment. Furthermore, polymorph SP delivers copper to cells such that copper is not released into the extracellular environment when administered to a patient.

[0019] The properties of CuATSM in polymorph SP are typically retained upon dissolution of the polymorph such that the intrinsic properties of CuATSM (including, but not limited to, cellular uptake, bioavailability, ability to cross the blood-brain barrier, redox potential, or therapeutic efficacy) are maintained.

[0020] definition Unless specifically stated otherwise herein, the definitions of the terms used are standard definitions used in the fields of organic synthesis and pharmaceutical sciences. Exemplary embodiments, aspects, and variations are illustrated in the illustrations and figures, and it is intended that the embodiments, aspects, and variations disclosed herein and the illustrations and figures be considered illustrative and non-limiting.

[0021] The term "neurodegenerative disorder" refers to a disorder in which the integrity of neurons is threatened. Neuronal integrity can be threatened when nerve cells show reduced viability or when neurons can no longer transmit signals. Neurological conditions that can be treated with compositions comprising the polymorph SP of the present application include the conditions listed herein.

[0022] The term "neurological condition" refers to a condition in which various cell types of the nervous system degenerate and / or are damaged as a result of a neurodegenerative disorder or injury or exposure. In particular, compositions containing the polymorph SP of the present application can be used to treat conditions resulting from damage to cells of the nervous system due to surgical intervention, infection, exposure to toxins, tumors, malnutrition, or metabolic disorders. Furthermore, compositions containing the polymorph SP can be used to treat sequelae of neurodegenerative disorders (such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, epilepsy, etc.), drug abuse or addiction (such as alcohol, cocaine, heroin, or amphetamines), spinal cord disorders, neuroretinal dystrophy or degeneration (retinopathy), and peripheral neuropathies (such as diabetic neuropathy and / or toxin-induced peripheral neuropathies).

[0023] As used herein, the term "patient" refers to any animal having a disease or condition requiring treatment or prevention with a biologically active agent. The patient may be a mammal (such as a human) or may be a non-human primate or non-primate used, for example, in animal model studies. While the compounds are suitable for use in human medical treatment, the compounds are also applicable to veterinary treatment.

[0024] The phrase "pharmaceutically acceptable" means that the compound, substance, or composition is chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the patient being treated.

[0025] The term "therapeutically effective amount" or "effective amount" is an amount sufficient to obtain beneficial or desired clinical results. An effective amount can be administered in one or more doses. An effective amount is typically sufficient to alleviate, ameliorate, stabilize, reverse, slow or retard the progression of a disease state.

[0026] In general, the terms "treatment" and "prevention" refer to affecting a subject, tissue, or cell to achieve a desired pharmacological and / or physiological effect, including: (a) preventing a condition from occurring in a subject who may be predisposed to the condition but has not yet been diagnosed as having the condition; (b) suppressing the condition (i.e., arresting its development); or (c) relieving or ameliorating the effects of the condition (i.e., reversing the effects of the condition).

[0027] Preparation method of CuATSM Methods for preparing metal complexes such as CuATSM and methods for treating various neurodegenerative diseases and disorders are disclosed in PCT / AU2007 / 001792, published as WO2008 / 061306, which is incorporated herein by reference in its entirety. Representative examples are provided herein.

[0028] Alternatively, CuATSM can be prepared using the reaction pathways and synthetic schemes below, using techniques available in the art for each individual step / reaction, and using readily available starting materials. Suitable protecting groups can be found in T.W. Greene's Protective Groups in Organic Synthesis, John Wiley & Sons, 1981.

[0029] The preparation of complexes such as CuATSM is shown in the following scheme: [ka]

[0030] Condensation of the dione (X) with two equivalents of a suitably functionalized thiosemicarbazide (XI) under acidic conditions forms the bis(thiosemicarbazone) (XIII), which can then be reacted with a suitable metal salt, such as a metal acetate, to produce the desired metal complex (XIV) and acetic acid.

[0031] Methods for preparing new polymorphs of CuATSM and gluconic acid The method for making the polymorph SP is described as follows: ATSMH2 (also called free ATSM ligand): [ka] (also known as (2E,2'E)-2,2'-(butane-2,3-diylidene)bis(N-methylhydrazine-1-carbothioamide) and copper gluconate: [ka] However, when mixed together, the resulting composition is a mixture of CuATSM and gluconic acid. In aqueous solution, gluconic acid is in equilibrium with its lactone form, gluconolactone. Gluconic acid exists in the (D) and (L) forms, as well as mixtures thereof (including racemic mixtures). In some embodiments, the resulting composition of polymorph SP is a mixture of CuATSM and D-gluconic acid.

[0032] The process for preparing polymorph SP may include the steps of mixing ATSMH2 and copper gluconate in a solvent to form a slurry; heating the slurry for a programmed period of time to form polymorph SP; and isolating the polymorph.

[0033] The above process can also be used to make polymorph SP starting with CuATSM and D-gluconic acid (instead of ATSMH2 and copper gluconate). Polymorph SP can also be made using the ball milling method as described in Example 1 below.

[0034] In the first step, ATSMH2 and copper gluconate are mixed together in a solvent to form a slurry. The ATSMH2 and copper gluconate are added in a molar ratio of between 2:1 and 1:2. In some embodiments, they are added in a molar ratio of between 1.05:1 and 1:1.05. In yet other embodiments, they are added in a molar ratio of about 1:1.

[0035] Solvents are C6-C 10 The solvent is selected from the group consisting of an alkane or cycloalkane, a C1-C6 alkyl acetate, and mixtures thereof. In some embodiments, the alkane or cycloalkane is hexane, heptane, or cycloheptane. In some embodiments, the alkyl acetate is methyl acetate, ethyl acetate, isopropyl acetate, or t-butyl acetate. In some embodiments, the solvent is heptane, isopropyl acetate, or a heptane / isopropyl acetate mixture.

[0036] The slurry is then slowly heated from 40° C. to 80° C. In some embodiments, the heating process is carried out in 20° C. increments over 15-25 days until the reaction is complete by HPLC analysis (ATSMH2 ≤ 2%).

[0037] The SP polymorph is then isolated. In some embodiments, isolation is achieved by filtration or centrifugation. In some embodiments, isolation is achieved by vacuum filtration followed by washing with heptane.

[0038] Methods of treatment, amelioration, and / or prevention Compositions containing polymorph SP are effective as copper metal delivery agents, particularly as agents for delivering copper to cells. Compositions containing polymorph SP can be used to treat or prevent several conditions that can be prevented, alleviated, or improved by metal delivery. There are several conditions of this type. Examples of conditions of this type are conditions related to or caused by oxidative stress. Because many biological antioxidant defense mechanisms involve copper-catalyzed enzymes, it is known that copper delivery can stimulate or restart the activity of biological antioxidant mechanisms, thereby exerting a comprehensive antioxidant effect. In one embodiment, the condition associated with or caused by oxidative stress is selected from the group consisting of cardiovascular conditions, cancer, cataracts, neurological disorders (such as Alzheimer's disease), prion diseases (including Creutzfeldt-Jakob disease (CJD)), and heart disease, amyloidogenic amyotrophic lateral sclerosis (ALS), prion transmissible spongioform encephalopathies (TSEs), cataracts, mitochondrial disorders, Menkes disease, Parkinson's disease, and Huntington's disease.

[0039] In another embodiment, the disorder is a neuromuscular disorder selected from the group consisting of amyotrophic lateral sclerosis (ALS), a mitochondrial / metabolic disease, and Friedreich's ataxia. In one embodiment, the symptom is a neurological symptom or a neurodegenerative disorder.

[0040] Additionally, compositions containing polymorph SP can be used to enhance the effects of other treatments (e.g., to enhance the neuroprotective effects of brain-derived nerve growth factor). Compositions containing polymorph SP can also be used to treat anemia, neutropenia, copper deficiency myelopathy, copper deficiency syndrome, and hyperzincemia. Additionally, treatment methods are directed to conditions that induce oxidative damage in the central nervous system, including acute and chronic neurological disorders such as cerebral ischemia, stroke (ischemic and hemorrhagic), subarachnoid hemorrhage / cerebral vasospasm, brain tumors, AD, CJD and its novel variants associated with "mad cow" disease, HD, PD, Friedreich's ataxia, cataracts, dementia with Lewy body formation, multiple system atrophy, Hallerboden-Spatz disease, diffuse Lewy body disease, amyotrophic lateral sclerosis, motor neuron disease, multiple sclerosis, fatal familial insomnia, Gertsmann-Straussler-Sheinker disease, and hereditary cerebral hemorrhage with amyloidosis of the Dutch type.

[0041] The treatment method is also directed to the treatment of neurodegenerative amyloidosis. Neurodegenerative amyloidosis can be any condition in which neuronal damage results from the deposition of amyloid. Amyloid can be formed from various protein or polypeptide precursors, including, but not limited to, Aβ, synuclein, huntingtin, or prion protein. In one embodiment, the condition is selected from the group consisting of sporadic or familial AD, ALS, motor neuron disease, cataracts, PD, Creutzfeldt-Jakob disease and its novel variants associated with "mad cow" disease, HD, dementia with Lewy body formation, multiple system atrophy, Hallerboden-Spatz disease, and diffuse Lewy body disease.

[0042] In another embodiment, the neurodegenerative amyloidosis is an Aβ-related condition, such as AD or dementia associated with Down's syndrome or one of several autosomal dominant forms of familial AD (reviewed in St. George-Hyslop, 2000). Most preferably, the Aβ-related condition is AD. In another embodiment, prior to treatment, the patient may have moderate or severe cognitive impairment as assessed by the AD Assessment Scale (ADAS)-cog test (e.g., an ADAS-cog score of 25 or greater). In addition to slowing or halting a subject's cognitive decline, compositions and methods comprising the polymorph SP of the present invention may also be suitable for use in treating or preventing neurodegenerative conditions, or for alleviating the symptoms of neurodegenerative conditions. When administered to patients identified as at high risk for a neurodegenerative condition or to subjects showing preclinical signs of cognitive decline (such as mild cognitive impairment or minimally progressing cognitive impairment), these compositions comprising polymorph SP and methods of use thereof may be able to prevent or delay the onset of clinical symptoms in addition to slowing or slowing the rate of cognitive decline.

[0043] In addition, compositions comprising the polymorph SP of the present application may be useful for the treatment of cancer. The term "cancer" describes any of a series of different diseases involving cumulative multigene mutations that activate oncogenes and / or inactivate tumor suppressor genes and / or contribute to uncontrolled cell proliferation. The causes and sources of these mutations vary among various cancers of the human body.

[0044] In one embodiment, the present application relates to a method of treating brain cancer (including brain tumors). Brain cancer or brain tumors can be gliomas or non-gliomatous brain tumors. As used herein, the terms "cancer" and "tumor" may be used interchangeably herein. "Cancer" can include any one of the following conditions: glioma, adenoma, blastoma, carcinoma, sarcoma, and also any one of medulloblastoma, ependymoma, astrocytoma, optic nerve glioma, brain stem glioma, oligodendroglioma, ganglioglioma, craniopharyngioma, or pineal region tumor. Reference to "glioma" includes GMB, astrocytoma, and anaplastic astrocytoma, or related brain cancers.

[0045] In addition, compositions containing the polymorph SP of the present application can be used to treat tau-related disorders. Tau protein is a protein expressed in the central nervous system and is an important protein because it plays a key role in neuronal architecture by stabilizing the intracellular microtubule network. Therefore, any impairment of the physiological role of tau protein, either due to shortening, hyperphosphorylation, or disruption of the balance between the six naturally occurring tau isoforms, is harmful to the subject, resulting in the formation of neurofibrillary tangles (NFTs), dystrophic neurites, and neuropil threads. The main protein subunit of these structures is microtubule-associated tau protein. The amount of NFTs found in autopsies of AD patients correlates with clinical symptoms, including intellectual decline. Therefore, tau protein plays an important role in AD pathology.

[0046] It is believed that the activity of the composition comprising the polymorph SP of the present application to reduce tau phosphorylation levels allows the delivery of metals to cells, thereby demonstrating its antioxidant activity. The fact that the complex acts as an antioxidant may mean that the complex provides desirable protection from OS, which can hyperphosphorylate tau and cause cellular dysfunction. As a result, these complexes can function as antioxidants due to their ability to deliver biologically important metals to cells (especially when oxidative stress is caused by metal deficiency), which means that the metal complexes may have the ability to prevent (or treat) tauopathies. There are several disorders or conditions that are recognized as tauopathies, or more colloquially, tauopathies. Disorders of this type include Richardson syndrome, progressive supranuclear palsy, argyrophilic grain disease, corticobasal degeneration, Pick's disease, frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), postencephalitic parkinsonism (PEP), dementia pugilistica, Down's syndrome, Alzheimer's disease, familial British dementia, familial Danish dementia, Parkinson's disease, Parkinson's disease complex of Guam (PDC), myotonic dystrophy, Hallevorden-Spatz disease, and Niemann-Pick disease type C.

[0047] Compositions containing polymorph SP can also be used in the treatment of Abeta-related disorders. Several Abeta disorders are known, including disorders selected from the group consisting of Parkinson's disease, Alzheimer's disease, multiple sclerosis, neuropathy, Huntington's disease, prion disease, motor neuron disease, amyotrophic lateral sclerosis (ALS), Menkes disease, and amyloidosis.

[0048] Compositions containing the SP polymorph have also been shown to be capable of delivering copper to cells, thereby affecting matrix metalloproteinases (MMPs). Matrix metalloproteinases (MMPs) are a family of zinc- and calcium-dependent secreted or membrane-anchored endopeptidases that perform several important biological functions. While MMPs are involved in numerous physiological processes, they are also implicated in the pathophysiological mechanisms underlying a wide range of diseases. Pathological expression and activation of MMPs have been linked to cancer, atherosclerosis, stroke, arthritis, periodontal disease, multiple sclerosis, and liver fibrosis.

[0049] In addition to slowing or halting cognitive decline in a subject, compositions and methods comprising the polymorph SP of the present invention may also be suitable for use in treating, preventing, or alleviating gastrointestinal (GI) diseases or disorders (such as constipation). When administered to patients identified as at high risk for neurodegenerative conditions and GI diseases or disorders, or to subjects showing preclinical evidence of cognitive decline and related GI diseases or disorders, these metal complexes and methods of use may be able to prevent or delay the onset of clinical symptoms as well as treat, prevent, or alleviate GI diseases or disorders, in addition to slowing or slowing the rate of cognitive decline. Although certain mechanisms of action are proposed herein, the inventors do not intend to be bound by any proposed or suggested mechanisms of action in the present invention.

[0050] In one embodiment, compositions comprising polymorph SP can be administered to mammals by oral or parenteral methods without the need for formulations with excipients, solubilizers, and the like that are not approved for human use.

[0051] Administration of Compositions Containing Polymorph SP The composition comprising polymorph SP can be administered to humans by any of the recognized administration modes well known in the art. For example, the composition can be administered enterally (such as orally or rectally) or parenterally (such as subcutaneously, intramuscularly, intravenously, and intradermally). The injection can be a bolus injection or a constant or intermittent infusion. The composition comprising polymorph SP typically contains a pharmaceutically acceptable carrier or diluent in an amount effective to deliver a therapeutically effective dose to the subject.

[0052] Compositions containing the SP polymorph can be administered in any form or manner that makes the complex bioavailable. Those skilled in the art of preparing formulations can easily select the appropriate form and manner of administration depending on the specific characteristics of the selected complex, the condition being treated, the stage of the condition being treated, and other relevant circumstances. Remington's Pharmaceutical Sciences, 19 th See, e.g., "The Art of Doping with Polymorph SP," ed., Mack Publishing Co. (1995). In one embodiment, the composition comprising polymorph SP can be administered alone or in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable carrier, diluent, or excipient.

[0053] Pharmaceutical compositions containing Polymorph SP for parenteral injection include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions immediately before use. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, and polyethylene glycol), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters (such as ethyl oleate). These compositions containing Polymorph SP can also contain adjuvants (such as preservatives, wetting agents, emulsifiers, and dispersing agents).

[0054] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active complex is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders (such as starch, lactose, sucrose, glucose, mannitol, and silicic acid), b) binders (such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia), c) humectants (such as glycerol), d) disintegrants (such as agar, calcium carbonate, jasmine, etc.), e) glycerol, f) glycerol, g) glycerol, h) glycerol, i) glycerol, j) glycerol, k ... The formulation may be mixed with any of the following additives: gum or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarders (such as paraffin); f) absorption accelerators (such as quaternary ammonium compounds); g) wetting agents (such as cetyl alcohol and glycerol monostearate); h) absorbents (such as kaolin and bentonite clay); and i) lubricants (such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents. [Example]

[0055] Example X-ray powder diffraction (XRPD) method The Rigaku Smart-Lab X-ray diffraction system was configured for reflection Bragg-Brentano geometry using an X-ray beam source. The X-ray source was a Cu Long Fine Focus tube operated at 40 kV and 44 mA. This source produced an incident beam profile at the sample that varied from a thin line at high angles to a wide rectangle at low angles. A beam-adjusting slit was used on the X-ray source to ensure that the maximum beam size was less than 10 mm both along and perpendicular to the line. The Bragg-Brentano geometry was a parallel-focusing type controlled by passive divergence and receiving slits, with the sample itself acting as the focusing component for the optics. The inherent resolution of the Bragg-Brentano geometry was controlled in part by the diffractometer radius and receiving slit width used. Typically, the Rigaku Smart-Lab was operated to obtain peak widths of 0.1° 2θ or less. The axial divergence of the X-ray beam was controlled by 5.0° Soller slits in both the incident and diffracted beam paths.

[0056] The samples were placed in a low-background silicon holder by gently pressing with a finger to keep the sample surface flat and level with the reference surface of the holder. Each sample was analyzed from 2 to 40° 2θ using a continuous scan of 6° 2θ / min with an effective step size of 0.02° 2θ.

[0057] Example 1 Synthesis of bulk powder of polymorph SP by ball milling A pre-weighed amount of ATSMH2 (1000.0 g) and copper(II) D-gluconate (1750.0 g) solids were placed in a PK blender. The solids were blended for 10 minutes. The resulting mixed powder was transferred to a ball mill equipped with a 10-liter porcelain jar (Shimpo) containing a 1-inch media ball. The ball mill was started to begin mixing, and mixing was continued, with frequent checks, until the copper turned dark brown. This occurred after 17.75 days of mixing. This process yielded 2368.8 g of product (86% yield, polymorph SP). The XRPD spectrum of this product is shown at the top of Figure 1 and is compared to the XRPD spectra of CuATSM, copper D-gluconate, D-gluconic acid lactone, and D-gluconic acid. The XRPD spectrum of polymorph SP shows characteristic peaks at approximately 7.5, 9, 11, 15.5, 27.5, 28.5, and 32 which do not appear in the XRPD spectra of CuATSM free base, copper D-gluconate, D-gluconic acid lactone, and D-gluconic acid.

[0058] Example 2 Synthesis of IPA slurry ATSMH2 (1 equivalent) and Cu(II) gluconate (1 equivalent) were suspended in isopropyl alcohol (20 vol) and stirred at 40 °C for 7 days, then at 60 °C for 8 days, and then at 80 °C for 2 days until the reaction was complete. The brown slurry was isolated by filtration, washed with heptane (2 × 2 volumes), and dried in a vacuum oven to give CuATSM gluconate (100% yield, not polymorph SP) as a brown solid. The XRPD spectrum of this product is shown at the top of Figure 2 and is compared with the XRPD spectra of CuATSM, copper D-gluconate, and D-gluconic acid lactone. As shown by XRPD, this method does not yield the CuATSM:gluconic acid polymorph with characteristic peaks at approximately 7.5, 9, 11, 15.5, 27.5, 28.5, and 32.

[0059] Example 3 Synthesis of heptane slurry of polymorph SP ATSMH2 (1 equivalent) and Cu(II) gluconate (1 equivalent) were suspended in heptane (20 vol) and stirred at 40 °C for 7 days, 60 °C for 8 days, and then heated at 80 °C for 2 days until the reaction was complete. The brown slurry was isolated by filtration, washed with heptane (2 × 2 volumes), and dried in a vacuum oven to give CuATSM gluconate (100% yield, polymorph SP) as a brown solid. The XRPD spectrum of this product is shown at the top of Figure 3 and is compared with the XRPD spectra of CuATSM, copper D-gluconate, and D-gluconic acid lactone. The XRPD spectrum of polymorph SP shows characteristic peaks at approximately 7.5, 9, 11, 15.5, 27.5, 28.5, and 32 which do not appear in the XRPD spectra of CuATSM free base, copper D-gluconate, D-gluconic acid lactone, and D-gluconic acid.

[0060] Example 4 Preparation of isopropyl acetate slurry of polymorph SP ATSMH2 (1 equiv.) and Cu(II) gluconate (1 equiv.) were suspended in isopropyl acetate (20 vol.) and stirred at room temperature for 8 days until the reaction was complete. The brown slurry was concentrated in vacuo (10 vol.), isolated by filtration, washed with isopropyl acetate (2 × 2 vol.), and dried in a vacuum oven to give CuATSM gluconate (94% yield, polymorph SP) as a brown solid. The XRPD spectrum of this product is shown at the top of Figure 4 and compared with the XRPD spectra of copper D-gluconate and D-gluconic acid. The XRPD spectrum of polymorph SP exhibits characteristic peaks at approximately 7.5, 9, 11, 15.5, 27.5, 28.5, and 32; these peaks do not appear in the XRPD spectra of CuATSM free base, copper D-gluconate, D-gluconic acid lactone, or D-gluconic acid.

[0061] Example 5 Synthesis of heptane / isopropyl acetate slurry of polymorph SP ATSMH2 (1 equivalent) and Cu(II) gluconate (1 equivalent) were suspended in heptane (15 volumes) and isopropyl acetate (15 volumes) and stirred at 40°C for 7 days, then heated at 60°C for 7 days, and then heated at 80°C for 4 days until the reaction was complete. The brown slurry was isolated by filtration, washed with heptane (2 × 2 volumes), and dried in a vacuum oven to give CuATSM gluconate (90% yield, polymorph SP) as a brown solid. The XRPD spectrum of this product contained characteristic 2θ peaks at angles of 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°; these peaks do not appear in the XRPD spectra of CuATSM free base, copper D-gluconate, D-gluconic acid lactone, and D-gluconic acid.

[0062] The XRPD spectra discussed above demonstrate that the products made in Examples 1, 3, 4, and 5 have characteristic spectra with 2θ peaks at angles of 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°, while the spectra of the product made in Example 2, the precursor materials CuATSM and copper D-gluconate (and, for that matter, D-gluconic acid and D-gluconic acid lactone) do not contain these peaks.

[0063] While several exemplary embodiments, aspects, and variations are provided herein, those skilled in the art will recognize certain modifications, permutations, additions, and combinations, and certain subcombinations, of the embodiments, aspects, and variations. It is intended that the following claims be interpreted to include all such modifications, permutations, additions, and combinations, and certain subcombinations, of the embodiments, aspects, and variations, and that they fall within the scope of the claims. The entire disclosures of all documents cited throughout this application are incorporated herein by reference. The present invention provides, for example, the following items. (Item 1) Cu II -diacetyl-bis(N 4 1. A stable polymorph of Cu(II)-methyl-thiosemicarbazone (CuATSM) and gluconic acid, wherein said composition has at least five XRPD spectral peaks selected from the group consisting of 2θ angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. (Item 2) 2. The polymorph according to claim 1, wherein the polymorph has at least six XRPD spectral peaks selected from the group consisting of 2θ angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. (Item 3) 3. The polymorph according to item 2, wherein the polymorph has peaks in an XRPD spectrum at 2θ angles of approximately 7.5°, 9°, 11°, 15.5°, 27.5°, 28.5°, and 32°. (Item 4) 10. A pharmaceutical composition comprising a therapeutically effective amount of the polymorph according to item 1, and a pharmaceutically acceptable excipient or salt. (Item 5) 10. A method for treating or preventing a condition in a mammal, wherein delivery of copper prevents, alleviates, or ameliorates said condition, comprising administering to said mammal a therapeutically effective amount of the composition of claim 4. (Item 6) The conditions include adriamycin-induced cardiomyopathy; AIDS dementia and HIV-1-induced neurotoxicity; Alzheimer's disease; acute intermittent porphyria; Alzheimer's disease (AD); amyotrophic lateral sclerosis (ALS); atherosclerosis; cataracts; cerebral ischemia; cerebral palsy; brain tumors; chemotherapy-induced organ damage; cisplatin-induced nephrotoxicity; coronary artery bypass surgery; Creutzfeldt-Jakob disease and its novel variants associated with "mad cow" disease; diabetic neuropathy; Down's syndrome; drowning; epilepsy and post-traumatic epilepsy; Friedreich's ataxia; frontotemporal dementia; glaucoma; glomerulopathy; hemochromatosis; hemodialysis; hemolytic uremic syndrome (Weil's disease); dementia with Lewy bodies, Menkes disease; hemorrhagic stroke; Hallerboden-Spatz disease disease); heart attack and reperfusion injury; Huntington's disease; Lewy body disease; intermittent claudication; ischemic stroke; inflammatory bowel disease; macular degeneration; malaria; methanol-induced toxicity; meningitis (aseptic and tuberculous); motor neuron disease; multiple sclerosis; multiple system atrophy; myocardial ischemia; neoplasia; Parkinson's disease; perinatal asphyxia; Pick's disease; progressive supranuclear palsy (PSP); radiation therapy-induced organ damage; restenosis after angioplasty; retinopathy; senile dementia; schizophrenia; sepsis; SCN2A-related The method of item 5, wherein the disease is selected from the group consisting of epileptic encephalopathy; septic shock; spongiform encephalopathy; subarachnoid hemorrhage / cerebral vasospasm; subdural hematoma; surgical trauma (including neurosurgery); thalassemia; transient ischemic attack (TIA); synucleinopathy; transplant; vascular dementia; viral meningitis; viral encephalitis; neuropathy, acrodermatitis enteropathica; dementia with Lewy bodies; tauopathy; mild cognitive impairment (MCI); motor neuron disease (MND), and prion disease. (Item 7) 7. The method of item 6, wherein the condition is a neurodegenerative disease selected from the group consisting of Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), Menkes disease, multiple sclerosis, neuropathy, motor neuron disease (MND), Parkinson's disease, Huntington's disease, frontotemporal dementia, acrodermatitis enteropathica, dementia with Lewy bodies, synucleinopathy, tauopathy, mild cognitive impairment (MCI), progressive supranuclear palsy (PSP), and prion disease. (Item 8) 1. A process for preparing stable polymorphs of CuATSM and gluconic acid, comprising: a.ATSMH 2 and mixing copper gluconate (in a first ratio) with a solvent (in a second ratio) to form a slurry; b. heating the slurry to form a composition comprising CuATSM and gluconic acid; and c. isolating said polymorph The process includes: (Item 9) 9. The process of claim 8, wherein the solvent is selected from the group consisting of heptane, isopropyl acetate, and a heptane / isopropyl acetate mixture. (Item 10) Item 10. The process of item 9, wherein the solvent is heptane. (Item 11) 10. The process of claim 9, wherein the solvent is a heptane / isopropyl acetate mixture. (Item 12) a.ATSMH 2 and mixing copper gluconate (in a first ratio) with a solvent (in a second ratio) to form a slurry; b. heating the slurry to form the composition; and c. isolating said polymorph 2. The polymorph of claim 1, produced by a process comprising: (Item 13) 13. The polymorph according to item 12, wherein the solvent is selected from the group consisting of heptane, isopropyl acetate, and a heptane / isopropyl acetate mixture. (Item 14) 14. The polymorph according to item 13, wherein the solvent is heptane. (Item 15) 14. The polymorph according to item 13, wherein the solvent is a heptane / isopropyl acetate mixture.

Claims

1. 1. A process for preparing a stable polymorph of a mixture of CuATSM and gluconic acid, comprising: ATSMH ２ and mixing copper gluconate with a solvent to form a slurry; b. heating the slurry to form a composition comprising CuATSM and gluconic acid; and c. isolating said polymorph wherein the solvent is selected from the group consisting of heptane, isopropyl acetate, and a heptane / isopropyl acetate mixture.

2. 10. The process of claim 1, wherein the solvent is heptane.

3. 10. The process of claim 1, wherein the solvent is a heptane / isopropyl acetate mixture.

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