Polymorphs and amorphous forms of compounds

Abstract

The present invention discloses polymorphic and amorphous forms of anhydrates, hydrates, and solvates of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, and methods for using such compositions to treat or inhibit oxidative stress disorders, including mitochondrial diseases, impaired energy processing disorders, neurodegenerative diseases, and aging diseases. The present invention further discloses methods for preparing such polymorphic and amorphous forms.

Description

[0001] The present invention claims priority to U.S. Provisional Patent Application No. 62 / 092,743, filed on December 16, 2014, entitled “POLYMORPHIC AND AMORPHOUS FORMS OF (R)-2-HYDROXY-2-METHYL-4-(2,4,5-TRIMETHYL-3,6-DIOXOCYCLOHEXA-1,4-DIENYL) BUTANAMIDE,” and U.S. Provisional Patent Application No. 62 / 133,276, filed on March 13, 2015, entitled “POLYMORPHIC AND AMORPHOUS FORMS OF (R)-2-HYDROXY-2-METHYL-4-(2,4,5-TRIMETHYL-3,6-DIOXOCYCLOHEXA-1,4-DIENYL) BUTANAMIDE,” both of which are incorporated herein by reference in their entireties for all purposes. Technical Field

[0002] The present application discloses compositions and methods for treating or inhibiting diseases, developmental delays and conditions associated with oxidative stress disorders. Examples of such diseases or conditions include mitochondrial diseases, impaired energy processing disorders, neurodegenerative diseases and aging diseases. The present application further discloses methods for preparing such compositions. Technical Background

[0003] Oxidative stress is caused by a disturbance in the normal redox state within cells. An imbalance between the regular production and detoxification of reactive oxygen species such as peroxides and free radicals can lead to oxidative damage to cellular structures and mechanisms. In aerobic organisms, the most important source of reactive oxygen species under normal conditions is probably the leakage of reactive oxygen species from mitochondria during normal oxidative respiration. Impairments associated with this process are suspected to contribute to mitochondrial diseases, neurodegenerative diseases, and diseases of aging.

[0004] Mitochondria are organelles in eukaryotic cells, often referred to as the "powerhouse" of the cell. One of its main functions is oxidative phosphorylation. The role of the adenosine triphosphate (ATP) molecule is to serve as the energy "currency" or energy carrier in the cell, and eukaryotic cells obtain most of their ATP from biochemical processes performed by the mitochondria. These biochemical processes include the citric acid cycle (tricarboxylic acid cycle, or Krebs cycle), which produces reduced nicotinamide adenine dinucleotide (NADH+H+) from oxidized nicotinamide adenine dinucleotide (NAD+), and oxidative phosphorylation, during which NADH+H+ is oxidized back to NAD+. The citric acid cycle also reduces flavin adenine dinucleotide, or FAD, to FADH2; FADH2 is also involved in oxidative phosphorylation.

[0005] The electrons released by NADH+H+ oxidation shuttle in a series of protein complexes (complex I, complex II, complex III and complex IV) known as the mitochondrial respiratory chain. These complexes are embedded in the inner membrane of the mitochondria. Complex IV, at the end of the chain, transfers the electrons to oxygen atoms, which are reduced to water. The energy released by these electrons passing through the complex is used to generate a proton gradient across the inner membrane of the mitochondria, which generates an electrochemical potential across the inner membrane. Another protein complex, complex V (which is not directly related to complexes I, II, III, and IV) uses the energy stored in the electrochemical gradient to convert ADP into ATP.

[0006] Anaerobic respiration is used when cells in an organism are temporarily deprived of oxygen until oxygen becomes available again or the cell dies. The pyruvate produced during glycolysis is converted to lactate during anaerobic respiration. When oxygen cannot be supplied to muscle cells, it is believed that the accumulation of lactate will lead to fatigue of the muscles during intense activity. When oxygen becomes available again, the lactate is converted back to pyruvate for oxidative phosphorylation.

[0007] Oxygen poisoning or oxygen toxicity is caused by high concentrations of oxygen which can be harmful to the body and increase the formation of free radicals and other structures such as nitric oxide, peroxynitrite and peroxytrioxide. Generally speaking, the body has many defense systems to protect against such damage, but at higher free oxygen concentrations, these systems are eventually overwhelmed over time and the rate of cell membrane damage exceeds the ability of the cell membrane systems to control or repair it. Cell damage and cell death then results.

[0008] Qualitative and / or quantitative disruption of oxygen delivery to tissues results in energetic disruption of red blood cell function and leads to various diseases such as hemoglobinopathies. A hemoglobinopathies is a genetic defect that results in an abnormal structure of one of the globin chains of the hemoglobin molecule. Common hemoglobinopathies include thalassemia and sickle cell disease. Thalassemia is an inherited autosomal recessive blood disorder. In thalassemia, a genetic defect results in a reduced rate of synthesis of one of the globin chains that make up hemoglobin. While thalassemia is a quantitative problem of synthesizing too few globins, sickle cell disease is a qualitative problem of synthesizing incorrectly functioning globins. Sickle cell disease is a blood disorder characterized by abnormal, rigid, and sickle-shaped appearance of red blood cells. The sickle shape reduces the flexibility of the cells and results in restricted movement through blood vessels, depriving downstream tissues of oxygen.

[0009] Mitochondrial dysfunction leads to various disease states. Some mitochondrial diseases are caused by mutations or deletions in the mitochondrial genome. If the threshold ratio of mitochondria in a cell is defective, and if the threshold ratio of such cells in a tissue has defective mitochondria, symptoms of tissue or organ dysfunction may result. In fact, any tissue can be affected, and a variety of symptoms may occur, depending on the extent to which different tissues are involved. Examples of some mitochondrial diseases include Friedreich's ataxia (FRDA), Leber's hereditary optic neuropathy (LHON), mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like attacks (MELAS), myoclonic seizures with broken red fiber syndrome (MERRF), Leigh syndrome (subacute necrotizing encephalopathy, Leigh Syndrome) and respiratory chain disorders. Most mitochondrial diseases involve children who show accelerated aging symptoms and signs, including neurodegenerative diseases, stroke, blindness, hearing impairment, visual impairment, diabetes, and heart failure.

[0010] Friedreich's ataxia is an autosomal recessive neurodegenerative and cardiometabolic disorder caused by decreased levels of the protein Frataxin. The disease results in a gradual loss of voluntary motor coordination (ataxia) and cardiac complications. Symptoms typically begin in childhood, and the disease progressively worsens as the patient ages; patients eventually become wheelchair-bound due to movement disorders.

[0011] Leber's hereditary optic neuropathy (LHON) is a disease characterized by blindness with an average age of onset between 27 and 34 years. Other symptoms, such as cardiac abnormalities and neurological complications, may also occur.

[0012] Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) can present in infants, children, or young adults. Stroke with vomiting and seizures is one of the most severe presentations; it is assumed that damage to mitochondrial metabolism in certain areas of the brain is responsible for cell death and neurological damage, rather than damage caused by blood loss as occurs with ischemic stroke.

[0013] Myoclonic seizures with broken red fibers syndrome (MERRF) is one of a class of rare muscle diseases called mitochondrial encephalopathies. Mitochondrial encephalopathy is a disease in which defects in the genetic material produce part of the cell structure that releases energy (mitochondria). This can lead to dysfunction of the brain and muscles (encephalomyopathy). The mitochondrial defects are always present, as well as "broken red fibers" (abnormalities of the tissue when viewed under a microscope). The most typical symptoms of MERRF syndrome are myoclonic seizures that are usually sudden, brief, and spasmodic, which may affect the limbs or the whole body, difficulty speaking (dysarthria), optic atrophy, short stature, hearing loss, dementia, and involuntary twitching of the eyes (nystagmus) may also occur.

[0014] Leigh syndrome (subacute necrotizing encephalopathy, Leigh Syndrome) is a rare hereditary nervous system metabolic disorder characterized by central nervous system degeneration, wherein the symptoms usually begin between 3 months and 2 years old at age, and develop rapidly. In most children, the first sign may be poor sucking ability, and loss of head control and motor skills. These symptoms may be accompanied by loss of appetite, vomiting, irritability, continuous crying and epileptic seizures. Along with the progress of the disease, symptoms may also include general weakness, lack of muscle tone and lactic acidosis attacks, which can cause respiratory and renal damage. Heart problems may also occur.

[0015] Coenzyme Q10 deficiency is a respiratory disorder with syndromes such as exercise intolerance and recurrent myoglobin myopathy manifested by ataxia, urinary incontinence, epileptic seizures or mental retardation, and renal failure (DiMauro et al., (2005) Neuromusc. Disord., 15:311-315, "Childhood-onset cerebellarataxia and cerebellar atrophy," (Masumeci et al., (2001)

[0016] Neurology 56:849-855 and Lamperti et al., (2003) 60:1206:1208)); and pediatric encephalopathy associated with renal disease. Biochemical assays of muscle homogenates from patients with coenzyme Q10 deficiency showed severe reductions in the activities of respiratory chain complexes I and II+III, while complex IV (COX) was moderately reduced (Gempel et al., (2007) Brain, 130(8):2037-2044).

[0017] Complex I deficiency or NADH dehydrogenase NADH-CoQ reductase deficiency is a respiratory chain disorder with symptoms that fall into three major forms: (1) a fatal pediatric multisystem disease characterized by developmental delay, muscle weakness, heart disease, congenital lactic acidosis, and respiratory failure; (2) a myopathy that begins in early childhood or adult life and manifests as exercise intolerance or weakness; and (3) a mitochondrial encephalomyopathy (including MELAS), which may begin in childhood or adult life and include a variable combination of symptoms and signs, including ophthalmoplegia, seizures, dementia, ataxia, hearing loss, pigmentary retinopathy, sensory neuropathy, and uncontrollable movements.

[0018] Complex II deficiency or succinate dehydrogenase deficiency is a respiratory chain disorder with symptoms including encephalomyopathy and a variety of manifestations including failure to thrive, developmental delay, hypotonia, lethargy, respiratory failure, ataxia, myoclonus, and lactic acidosis.

[0019] Complex III deficiency or ubiquinone-cytochrome c oxidoreductase deficiency is a respiratory chain disorder with symptoms classified into four major forms: (1) fatal pediatric encephalomyopathy, congenital lactic acidosis, hypotonia, dystrophic posture, seizures, and coma; (2) late-onset encephalomyopathy (infant to adult): weakness, short stature, ataxia, dementia, hearing loss, sensory neuropathy, pigmentary retinopathy, and various combinations of pyramidal tract signs; (3) myopathy, with exercise intolerance modified by focal weakness; and (4) pediatric histiocytoid cardiomyopathy.

[0020] Complex IV deficiency, or cytochrome c oxidase deficiency, is a respiratory chain disorder with symptoms that fall into two major forms: (1) an encephalomyopathy in which patients are typically normal for the first 6 to 12 months of life and then show developmental regression, ataxia, lactic acidosis, optic atrophy, ophthalmoplegia, nystagmus, dystonia, pyramidal signs, respiratory problems, and frequent seizures; and (2) a myopathy with two major variants: (a) a fatal pediatric myopathy that may develop shortly after birth with hypotonia, weakness, lactic acidosis, shredded red fibers, respiratory failure, and renal problems; and (b) a benign pediatric myopathy that may develop shortly after birth with hypotonia, weakness, lactic acidosis, shredded red fibers, respiratory problems, but (if the child survives) then improves spontaneously.

[0021] Complex V deficiency or ATP synthase deficiency is a respiratory chain disorder that includes symptoms such as slowly progressive myopathy.

[0022] CPEO or chronic progressive external ophthalmoplegia syndrome is a respiratory chain disorder that includes symptoms such as visual myopathy, retinitis pigmentosa, or central nervous system dysfunction.

[0023] Kearns-Sayre syndrome (KSS) is a mitochondrial disease characterized by a triad of symptoms including: (1) typical onset in people younger than 20 years of age; (2) chronic, progressive, extraocular muscle palsies; and (3) retinitis pigmentosa. In addition, KSS may include cardiac conduction defects, cerebellar ataxia, and elevated cerebrospinal fluid (CSF) protein levels (e.g., >100 mg / dL). Other features associated with KSS may include myopathy, dystonia, endocrine dysfunction (e.g., diabetes, growth retardation or short stature, hypoparathyroidism), bilateral sensorineural hearing loss, dementia, cataracts, and proximal renal tubular acidosis.

[0024] Maternally inherited diabetes and deafness (MIDD) is a mitochondrial disorder characterized by maternally transmitted diabetes and sensorineural deafness. In most cases, MIDD is caused by point mutations in the mitochondrial gene MT-TL1, encoding the mitochondrial tRNA for leucine, and in rare cases, in the MT-TE and MT-TK genes, encoding the mitochondrial tRNAs for glutamate and lysine, respectively.

[0025] In addition to congenital diseases involving inherited defective mitochondria, acquired mitochondrial dysfunction leads to disease, particularly neurodegenerative diseases associated with aging, such as Parkinson's disease, Alzheimer's disease, and Huntington's disease. The incidence of somatic mutations in mitochondrial DNA increases exponentially with age; reduced respiratory chain activity is common in the aging population. Mitochondrial dysfunction has also been implicated in excitability, neuronal damage, such as that associated with cerebrovascular accidents, epileptic seizures, and ischemia.

[0026] Some of the above diseases appear to be caused by defects in complex I of the respiratory chain. The transfer of electrons from complex I to the rest of the respiratory chain is mediated by the compound coenzyme Q (also known as ubiquinone). Oxidized coenzyme Q (CoQ 氧化 or ubiquinone) is reduced by complex I to reduced coenzyme Q (CoQ 还原 Then, reduced coenzyme Q transfers its electrons to complex III of the respiratory chain, where the reduced coenzyme Q is oxidized again to CoQ 氧化 (ubiquinone). Then CoQ 氧化 can participate in further iterations of electron transfer.

[0027] Very few treatments are available for patients suffering from these mitochondrial diseases. Recently, the compound idebenone has been proposed for the treatment of Friedreich's ataxia. Although the clinical effects of idebenone have been relatively modest, the complications of mitochondrial diseases can be so severe that even a mildly useful treatment is preferable to untreated disease. Another compound, MitoQ, has been proposed for the treatment of mitochondrial diseases (see, U.S. Pat. No. 7,179,928); clinical results for MitoQ have not been reported. Administration of coenzyme Q10 (CoQ10) and vitamin supplements has shown only transient beneficial effects in individual cases of KSS. CoQ10 supplements have also been used to treat CoQ10 deficiency with inconsistent results.

[0028] Oxidative stress is suspected to be important in neurodegenerative diseases such as motor neuron disease, amyotrophic lateral sclerosis (ALS), Creutzfeldt-Jakob disease, Machado-Joseph disease, spinocerebellar ataxia, multiple sclerosis (MS), Parkinson's disease, Alzheimer's disease, and Huntington's disease. Oxidative stress is thought to be involved in certain cardiovascular diseases and also plays a role in the ischemic cascade due to oxygen reperfusion injury following hypoxia. The cascade includes both stroke and heart attack.

[0029] The damage accumulation theory, also known as the free radical theory of aging, evokes the random action of free radicals produced during aerobic metabolism, which causes damage to DNA, lipids, and proteins and accumulates over time. The concept of free radicals playing a role in the aging process was first introduced in Himan D (1956), "Aging-A theory based on free-radical and radiation chemistry," J. Gerontol. 11, 298-300.

[0030] According to the free radical theory of aging, the aging process begins with oxygen metabolism (Valko et al, (2004), "Role of oxygen radicals in DNA damage and cancer incidence," Mol. Cell. Biochem., 266, 37-56). Even under ideal conditions, some electrons can "leak" from the electron transport chain. These leaked electrons interact with oxygen to produce superoxide radicals, so that under physiological conditions, about 1-3% of oxygen molecules in mitochondria are converted into superoxide. The main site of free radical oxygen damage from superoxide radicals is mitochondrial DNA (mtDNA) (Cadenasetal., (2000) Mitochondrial free radical generation, oxidative stress and aging, Free Radic. Res, 28, 601-609). Cells repair a large amount of damage to nuclear DNA (nDNA), but mtDNA repair seems to be less efficient. Therefore, extensive mtDNA damage accumulates over time and shuts down mitochondria, leading to cell death and aging of the organism.

[0031] Some of the diseases associated with aging are cancer, diabetes, hypertension, atherosclerosis, ischemia / reperfusion injury, rheumatoid arthritis, neurodegenerative diseases such as dementia, Alzheimer's disease and Parkinson's disease. Diseases or symptoms caused by the physiological decline process of aging include decreased muscle strength, cardiopulmonary function, vision and hearing, as well as wrinkled skin and graying hair.

[0032] The ability to regulate the biological production of energy has applications beyond the above-mentioned diseases. Various other diseases can result in suboptimal levels of energy biomarkers (sometimes also referred to as energy function indicators), such as ATP levels. There is also a need to treat these diseases in order to regulate one or more energy biomarkers to improve patient health. In other applications, it may be necessary to regulate certain energy biomarkers away from their normal values ​​in individuals who do not suffer from the disease. For example, if an individual is undergoing a very difficult task, it is desirable to increase the ATP level of the individual.

[0033] Certain polymorphic or amorphous forms of a drug may have more advantageous characteristics than other forms; for example, improved stability, increased solubility, better handling properties, lack of associated toxic solvents, and increased purity.

[0034] PCT application number is PCT / US2008 / 082374, which was published on May 14, 2009 as Example 16 in WO 2009 / 061744, which describes the synthesis of racemic 2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide; however, this example does not specifically describe the synthesis of any specific polymorph or amorphous form of 2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide or any specific stereoisomer thereof. Summary of the Invention

[0035] One aspect of the present invention relates to a polymorph of an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is selected from the group consisting of Form I, Form II, Form III, Form IV, Form V, and Form VI described in the present invention.

[0036] Another aspect of the present invention relates to a polymorph of anhydrous (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form I as described herein. In some embodiments, the polymorph has substantially Fig.10The X-ray powder diffraction pattern shown. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 17.03, and 7.26. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 17.03, and 17.26. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, and 17.26. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 7.67, 10.75, 12.06, 15.33, 16.41, 17.03, 17.26, 18.72, 20.04, and 23.92. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 7.67, 10.75, 12.06, 15.33, 16.41, 17.03, 17.26, 18.72, 20.04, 20.64, 20.91, 21.14, 22.58, 23.13, 23.92, 24.19, 24.53, 27.21, and 27.56. In some embodiments, the polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 5.48, 7.67, 10.75, 12.06, 15.33, 16.41, 17.03, 17.26, 17.71, 17.94, 18.40, 18.72, 19.51, 20.04, 20.64, 20.91, 21.14, 21.55, 21.91, 22.25, 22.58, 23.13, 23.41, 23.92, 24.19, 24.53, 25.64, 26.13, 26.34, 27.21, 27.56, 28.01, 29.04, and 29.46. In some embodiments, including any of the preceding embodiments, the angular position may vary by ±0.1.In some embodiments, including any of the foregoing embodiments, the angular position can vary by ±0.05. In some embodiments, including any of the foregoing embodiments, the angular position can vary by ±0.02. In some embodiments, including any of the foregoing embodiments, the polymorphs are isolated. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein the composition is substantially free of Forms II-VI, wherein Forms II-VI are described in Table A or Tables 3-7, respectively. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein at least about 95 mole % of the composition is the polymorph, excluding any solvent, carrier, or excipient.

[0037] Another aspect of the present invention relates to a polymorph of a solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form V as described herein. In some embodiments, the polymorph has substantially Fig.30The X-ray powder diffraction pattern shown in a) or b) of . In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, and 15.45. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, and 15.45. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 15.45, and 23.96. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 14.80, 15.45, and 23.96. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 14.80, 15.45, 16.53, 23.96, 24.54, and 26.05. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 14.80, 15.45, 16.10, 16.34, 16.53, 20.18, 22.52, 22.86, 23.96, 24.54, and 26.05. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 6.91, 7.72, 9.61, 11.49, 11.86, 12.93, 13.19, 13.87, 14.80, 15.45, 16.10, 16.34, 16.53, 17.14, 17.85, 19.12, 19.85, 20.18, 21.00, 22.06, 22.52, 22.86, 23.09, 23.96, 24.54, 25.26, 26.05, and 26.90. In some embodiments, including any of the foregoing embodiments, the angular positions may vary by ±0.1. In some embodiments, including any of the foregoing embodiments, the angular positions may vary by ±0.05. In some embodiments, including any of the preceding embodiments, the angular position can vary by ±0.02.In some embodiments, including any of the preceding embodiments, the polymorph is isolated. In some embodiments, including any of the preceding embodiments, the polymorph is present in a composition, wherein the composition is substantially free of Forms I-IV and VI, wherein Forms I-IV and VI are described in Table A or Table 2 or Tables 4-7, respectively. In some embodiments, including any of the preceding embodiments, the polymorph is present in a composition, wherein at least about 95 mole % of the composition is the polymorph, not counting any solvent, carrier, or excipient.

[0038] Another aspect of the present invention relates to a polymorph of a hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form III of the present invention. In some embodiments, the polymorph has substantially Fig. 20The X-ray powder diffraction pattern shown in a) or b) of . In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 14.02, 15.23, and 21.10. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 14.02, 15.23, and 21.10. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 14.02, 15.23, and 21.10. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 13.74, 14.02, 15.23, and 21.10. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 14.02, 15.23, 21.10, and 22.69. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.81, 13.74, 14.02, 15.23, 21.10, 22.69, and 23.90. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.81, 13.74, 14.02, 15.23, 17.35, 21.10, 22.69, 23.15, 23.90, and 26.10. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.53, 11.81, 12.68, 12.93, 13.74, 14.02, 15.23, 16.53, 17.35, 17.98, 18.54, 19.09, 20.23, 21.10, 21.93, 22.69, 23.15, 23.50, 23.90, 24.65, 25.09, 25.46, 25.79, 26.10, 27.79, 28.22, 28.93, and 29.33. In some embodiments, including any of the preceding embodiments, the angular positions may vary by ±0.1. In some embodiments, including any of the preceding embodiments, the angular position may vary by ±0.05. In some embodiments, including any of the preceding embodiments, the angular position may vary by ±0.02.In some embodiments, including any of the preceding embodiments, the polymorph is isolated. In some embodiments, including any of the preceding embodiments, the polymorph is present in a composition, wherein the composition is substantially free of Forms I, II, IV, V, and VI, wherein Forms I, II, IV, V, and VI are described in Table A or Tables 2-3 and Tables 5-7, respectively. In some embodiments, including any of the preceding embodiments, the polymorph is present in a composition, wherein at least about 95 mole % of the composition is the polymorph, not counting any solvent, carrier, or excipient.

[0039] Another aspect of the present invention relates to a polymorph of anhydrous (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form II as described herein. In some embodiments, the polymorph has substantially Fig.15a), or b). In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 11.33, and 19.33. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 11.33, and 19.33. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 11.33, 10.85, and 19.33. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 11.33, 10.85, 19.33, and 17.3. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 10.85, 11.33, 13.47, and 19.33. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 5.76, 8.04, 9.63, 10.85, 11.33, 11.97, 13.47, 14.75, 17.37, 17.71, and 19.33. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 5.76, 8.04, 9.63, 10.85, 11.33, 11.97, 13.47, 14.75, 16.42, 16.89, 17.37, 17.71, 19.33, 22.89, and 24.59. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 5.76, 6.72, 7.57, 8.04, 9.63, 10.85, 11.33, 11.97, 12.38, 13.13, 13.47, 14.75, 15.28, 16.42, 16.89, 17.37, 17.71, 18 .17, 18.66, 19.33, 20.01, 20.29, 20.67, 20.90, 21.36, 21.54, 21.80, 22.55, 22.89, 23.27, 23.54, 23.87, 24.35, 24.59, 24.87, 25.29, 25.55, 25.89, 26.44, 27.49, 28.01, 28.39, and 29.17.In some embodiments, including any of the foregoing embodiments, the angular position may vary by ±0.1. In some embodiments, including any of the foregoing embodiments, the angular position may vary by ±0.05. In some embodiments, including any of the foregoing embodiments, the angular position may vary by ±0.02. In some embodiments, including any of the foregoing embodiments, the polymorphs are isolated. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein the composition is substantially free of Forms I, and III-VI, wherein Forms I, and III-VI are described in Table A or Tables 2-4 and Tables 6-7, respectively. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein at least about 95 mole % of the composition is the polymorph, excluding any solvent, carrier, or excipient.

[0040] Another aspect of the present invention relates to a polymorph of a solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form IV of the present invention. In some embodiments, the polymorph has substantially Fig.25The X-ray powder diffraction pattern shown in a), b) or c) of . In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 12.97, and 13.20. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 12.97, and 13.20. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, and 13.20. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 7.94, 8.76, 12.97, 13.20, 16.66, 17.33, and 20.57. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 7.94, 8.76, 12.97, 13.20, 15.08, 16.66, 17.33, 19.09, 20.57, and 21.58. In some embodiments, the polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 5.77, 6.28, 7.53, 7.94, 8.76, 9.39, 9.87, 10.54, 11.07, 11.68, 12.02, 12.28, 12.97, 13.20, 13.52, 14.40, 15.08, 15.90, 16.66, 16.96, 17.33, 17.59, 18.77, 19.09, 19.74, 20.27, 20.57, 21.09, 21.58, 22.81, 23.23, 24.01, 24.65, and 25.60. In some embodiments, including any of the preceding embodiments, the angular position may vary by ±0.1. In some embodiments, including any of the preceding embodiments, the angular position may vary by ±0.05.In some embodiments, including any of the foregoing embodiments, the angular position can vary by ±0.02. In some embodiments, including any of the foregoing embodiments, the polymorphs are isolated. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein the composition is substantially free of Forms I-III and V-VI, wherein Forms I-III and V-VI are described in Table A or Tables 2-5 and Table 7, respectively. In some embodiments, including any of the foregoing embodiments, the polymorphs are present in a composition, wherein at least about 95 mole % of the composition is the polymorph, not counting any solvent, carrier, or excipient.

[0041] Another aspect of the present invention relates to a polymorph of a solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form VI of the present invention. In some embodiments, the polymorph has substantially Fig.33The X-ray powder diffraction pattern shown in a) of . In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.91, and 12.94. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.91, and 12.94. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.91, 12.94, and 15.71. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at least at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.91, 12.94, 15.71, and 19.13. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 13.29. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 8.85, 9.41, 9.91, 12.94, 13.29, 16.67, and 19.13. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.39, 6.27, 8.85, 9.41, 9.91, 11.32, 12.94, 13.29, 14.03, 16.67, 19.13, 20.76, and 22.06. In some embodiments, the X-ray powder diffraction pattern of the polymorph comprises characteristic peaks at the following angular positions, wherein the angular positions may vary by ±0.2: 4.39, 6.27, 7.00, 8.62, 8.85, 9.41, 9.91, 11.32, 11.50, 12.25, 12.56, 12.94, 13.29, 14.03, 14.82, 15.10, 15.44, 15.71, 16.01, 16.67, 16.91, 17.33, 17.59, 18.33, 18.75, 19.13, 20.25, 20.76, 21.68, 22.06, 22.27, 22.61, 22.94, 24.01, 24.33, 24.65, 25.48, 26.05, 28.63, and 29.18. In some embodiments, including any of the preceding embodiments, the angular position can vary by ±0.1. In some embodiments, including any of the preceding embodiments, the angular position can vary by ±0.05.In some embodiments, including any of the foregoing embodiments, the angular position can vary by ±0.02. In some embodiments, including any of the foregoing embodiments, the polymorph is isolated. In some embodiments, including any of the foregoing embodiments, the polymorph is present in a composition, wherein the composition is substantially free of Form IV, wherein Form IV is described in Table A or Tables 2-6, respectively. In some embodiments, including any of the foregoing embodiments, the polymorph is present in a composition, wherein at least about 95 mole % of the composition is the polymorph, not counting any solvent, carrier, or excipient.

[0042] Another aspect of the invention relates to a composition comprising amorphous (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. In some embodiments, the amorphous form is isolated. In some embodiments, the composition is substantially free of Forms I-VI, wherein Forms I-VI are described in Table A or Tables 2-7, respectively. In some embodiments, including any of the preceding embodiments, at least about 95% of the composition is amorphous (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, excluding any solvent, carrier or excipient.

[0043] Another aspect of the present invention relates to a pharmaceutical composition comprising a polymorphic or amorphous form of an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, or a composition comprising this form as described herein, including any of the foregoing or later embodiments, and a pharmaceutically acceptable carrier. In some embodiments, the form is a Type I polymorph. In some embodiments, the form is a Type II polymorph. In some embodiments, the form is a Type III polymorph. In some embodiments, the form is a Type IV polymorph. In some embodiments, the form is a Type V polymorph. In some embodiments, the form is a Type VI polymorph. In some embodiments, the form is amorphous. In some embodiments, the pharmaceutical composition has an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a HPLC purity greater than about 95%, excluding any solvent, carrier, or excipient. In some embodiments, the pharmaceutical composition has an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a HPLC purity greater than about 99%, excluding any solvent, carrier, or excipient. In some embodiments, the pharmaceutical composition has an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide having an HPLC purity greater than about 99.9%, excluding any solvent, carrier, or excipient. HPLC purity % refers to the ratio of the HPLC peak area of ​​a given compound to the total peak area in a given HPLC spectrum. HPLC % is calculated by dividing the compound peak area by the total peak area in the HPLC spectrum and multiplying the quotient by one hundred.

[0044] Another aspect of the present invention relates to a pharmaceutical composition comprising an active drug and a pharmaceutically acceptable carrier, wherein the active drug comprises, or substantially comprises, a polymorph or amorphous form of an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide of the present invention. In some embodiments, the form is a type I polymorph. In some embodiments, the form is a type II polymorph. In some embodiments, the form is a type III polymorph. In some embodiments, the form is a type IV polymorph. In some embodiments, the form is a type V polymorph. In some embodiments, the form is a type VI polymorph. In some embodiments, the form is amorphous.

[0045] Another aspect of the present invention relates to a method for treating or inhibiting oxidative stress disorder, regulating one or more energy biomarkers, normalizing one or more energy biomarkers, or enhancing one or more energy biomarkers, the method comprising administering to an individual in need thereof a therapeutically effective amount or an effective amount of a polymorphic or amorphous form of an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, or a composition comprising such a form as described herein, including any of the preceding or following embodiments. The method may use any single polymorphic or amorphous form described herein, or a combination thereof. In some embodiments, the form is a Type I polymorph. In some embodiments, the form is a Type II polymorph. In some embodiments, the form is a Type III polymorph. In some embodiments, the form is a Type IV polymorph. In some embodiments, the form is a Type V polymorph. In some embodiments, the form is a Type VI polymorph. In some embodiments, the form is amorphous. In some embodiments, including any of the preceding embodiments, the anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is administered in a pharmaceutical composition comprising the polymorphic or amorphous form and a pharmaceutically acceptable carrier. In some embodiments, including any of the preceding embodiments, the pharmaceutical composition comprises an active ingredient that is essentially a polymorphic or amorphous form of the anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. In some embodiments, including any of the preceding embodiments, the method is a method of treating or inhibiting oxidative stress disorders. In some embodiments, including any of the preceding embodiments, the method is a method of treating oxidative stress disorders. In some embodiments, including any of the preceding embodiments, the method is a method of inhibiting oxidative stress disorders. In some embodiments, including any of the preceding embodiments,The oxidative stress disorder is selected from the group consisting of: mitochondrial disease; inherited mitochondrial disease; Alpers disease; Barth syndrome; beta oxidation defect; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive external ophthalmoplegia (CPEO); CPT I deficiency; CPT II deficiency; Friedreich's ataxia (FA); glutaric aciduria type II; Kearns-Sayre syndrome (KSS); lactic acidosis; long chain acyl-CoA dehydrogenase deficiency (LCAD); LCHAD; Leigh syndrome (subacute necrotizing encephalopathy, Leigh Syndrome; Leigh-like syndrome; Leber's hereditary optic neuropathy (LHON); fatal infantile cardiomyopathy (LIC); Luft disease; multiple acyl-CoA dehydrogenase deficiency (MAD); medium-chain acyl-CoA dehydrogenase deficiency (MCAD); mitochondrial myopathy, encephalopathy, lactic acidosis, stroke-like episodes (MELAS); myoclonic seizures with ragged red fibers (MERRF); mitochondrial recessive ataxia syndrome (MIRAS); mitochondrial cytopathy, mitochondrial DNA depletion; mitochondrial encephalopathy; mitochondrial myopathy; mitochondrial neurogastrointestinal encephalopathy (MNGIE); neuropathy, ataxia,and retinitis pigmentosa (NARP); Pearson syndrome; pyruvate carboxylase deficiency; pyruvate dehydrogenase deficiency; POLG mutations; respiratory chain disorders; short-chain acyl-CoA dehydrogenase deficiency (SCAD); SCHAD; very long-chain acyl-CoA dehydrogenase deficiency (VLCAD); myopathy; cardiomyopathy; encephalomyopathy; neurodegenerative diseases; Parkinson's disease; Alzheimer's disease; amyotrophic lateral sclerosis (ALS); motor neuron disease; nervous system diseases; epilepsy; age-related diseases; macular degeneration; diabetes; metabolic syndrome; cancer; brain cancer; genetic diseases; Huntington's disease; mood disorders; schizophrenia; bipolar disorder; pervasive developmental disorder; autism; Asperger's syndrome; childhood disintegrative disorder (CDD); Rett's disorder; PDD-NOS; cerebrovascular accident; stroke; visual impairment; optic neuropathy; dominant juvenile optic atrophy; optic neuropathy induced by toxic drugs; glaucoma; Stargardt's macular dystrophy; diabetic retinopathy; diabetic maculopathy; retinopathy of prematurity; retinal damage associated with ischemia-reperfusion; oxygen toxicity; hemoglobinopathies; thalassemia; sickle cell anemia; seizures; ischemia; renal tubular acidosis; attention-deficit / hyperactivity disorder (ADHD); neurodegenerative disorders causing hearing or balance disorders; dominant optic atrophy (DOA); maternally inherited diabetes and deafness (MIDD); chronic fatigue; contrast-induced renal damage; contrast-induced retinopathy damage; abetalipoproteinemia; retinal atrophy; Wolfram disease; Tourette syndrome syndrome); cobalamin C deficiency; methylmalonic aciduria; glioblastoma; Down syndrome; acute tubular necrosis; muscular dystrophy; leukodystrophy; progressive supranuclear palsy; spinal muscular atrophy; hearing loss; noise-induced hearing loss; traumatic brain injury; juvenile Huntington's disease; multiple sclerosis; NGLY1; multiple system atrophy; adrenoleukodystrophy; and adrenomyeloneuropathy. In some embodiments, the oxidative stress disorder is multiple system atrophy. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is cancer. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is bipolar disorder. In some embodiments, the oxidative stress disorder is schizophrenia. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is an age-related disease. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Huntington's disease. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Alzheimer's disease. In some embodiments,Including any of the preceding embodiments, the oxidative stress disorder is amyotrophic lateral sclerosis (ALS). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is epilepsy. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Parkinson's disease. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is epileptic seizures. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is stroke. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is mitochondrial myopathy. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is a hereditary mitochondrial disease. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Friedreich's ataxia (FA). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Kearns-Sayre syndrome (KSS). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Leigh syndrome (subacute necrotizing encephalopathy, Leigh Syndrome) or Leigh-like syndrome. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Leber's hereditary optic neuropathy (LHON). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is mitochondrial myopathy, encephalopathy, lactic acidosis, stroke-like episodes (MELAS). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is myoclonic seizures with broken red fibers (MERRF). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is macular degeneration. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is brain cancer. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is autism. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is Rett's disorder. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is maternal inherited diabetes and deafness (MIDD). In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is chronic fatigue. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is contrast-induced renal injury. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is contrast-induced retinopathy injury. In some embodiments, including any of the preceding embodiments, the oxidative stress disorder is cobalamin C deficiency. In some embodiments, including any of the preceding embodiments, the method is to modulate one or more energy biomarkers, normalize one or more energy biomarkers,or a method of enhancing one or more energy biomarkers, wherein the one or more energy biomarkers are selected from the group consisting of: lactate (lactate) levels in whole blood, plasma, cerebrospinal fluid, or in ventricular fluid; pyruvate (pyruvate) levels in whole blood, plasma, cerebrospinal fluid, or in ventricular fluid; lactate / pyruvate / lactate ratios in whole blood, plasma, cerebrospinal fluid, or in ventricular fluid; total, reduced or oxidized glutathione levels or reduced / oxidized glutathione ratios in whole blood, plasma, cerebrospinal fluid, or in ventricular fluid; total, reduced or oxidized cysteine ​​levels or reduced / oxidized cysteine ​​ratios in whole blood, plasma, cerebrospinal fluid, or in ventricular fluid; creatine phosphate levels, NADH (NADH+H+) levels; NADPH (NADPH+H+) levels; NAD levels; NADP levels; ATP levels; reduced coenzyme Q (CoQ, 还原 ) level; oxidized coenzyme Q (CoQ 氧化 ) level; total coenzyme Q (CoQ 总) levels; oxidized cytochrome C levels; reduced cytochrome C levels; oxidized cytochrome C / reduced cytochrome C ratio; acetoacetate / root level, β-hydroxypyruvate / root level, acetoacetate / root / β-hydroxypyruvate / root ratio, 8-hydroxy-2'-deoxyguanosine (8-OHdG) level; reactive oxygen species level; oxygen consumption level (VO2); carbon dioxide output level (VCO2); respiratory quotient (VCO2 / VO2); exercise tolerance; and anaerobic threshold. Energy biomarkers can be measured in whole blood, plasma, cerebrospinal fluid, ventricular fluid, arterial blood, venous blood, or any other body fluid, body gas, or other biological sample used for such measurement. In some embodiments, including any of the preceding embodiments, the levels are adjusted to values ​​within about 2 standard deviations of the values ​​of healthy subjects. In some embodiments, including any of the preceding embodiments, the levels are adjusted to values ​​within about 1 standard deviation of the values ​​of healthy subjects. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 10% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 20% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 30% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 40% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 50% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 75% above or below the subject's level. In some embodiments, including any of the foregoing embodiments, prior to adjustment, the level has a change of at least about 100% above or at least about 90% below the subject's level.In some embodiments, including any of the preceding embodiments, the subject or object for which the method of treating or inhibiting oxidative stress disorder, modulating one or more energy biomarkers, normalizing one or more energy biomarkers, or enhancing one or more energy biomarkers is performed is a subject / subject selected from the group consisting of: a subject undergoing strenuous or prolonged physical activity; a subject with chronic energy problems; a subject with chronic respiratory problems; a pregnant woman; a pregnant woman in labor; a newborn; a premature newborn; a subject exposed to extreme environments; a subject exposed to heat; a subject exposed to cold; a subject exposed to below average oxygen levels; a subject exposed to above average carbon dioxide levels; a subject exposed to above average levels of air pollution. subjects in environments with reduced air quality; airline travelers; flight personnel; subjects at high altitudes; urban populations with below-average urban air quality; persons working in enclosed environments with reduced air quality; patients with lung disease; subjects with below-average lung capacity; patients with tuberculosis; patients with lung cancer; patients with emphysema; patients with cystic fibrosis; patients recovering from surgery; persons recovering from illness; the elderly; the elderly experiencing energy loss; subjects suffering from chronic fatigue; subjects suffering from chronic fatigue syndrome; subjects with acute trauma; subjects in shock; subjects requiring acute oxygen therapy; subjects requiring chronic oxygen therapy; subjects requiring contrast solutions for visualization of organs; or other subjects with acute, chronic, or ongoing energy needs who may benefit from enhanced energy biomarkers.

[0046] Another aspect of the present invention relates to the use of a polymorph or amorphous form of the anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide of the present invention, including any of the foregoing or subsequent embodiments, for treating or preventing oxidative stress disorders. In some embodiments, the form is a Type I polymorph. In some embodiments, the form is a Type II polymorph. In some embodiments, the form is a Type III polymorph. In some embodiments, the form is a Type IV polymorph. In some embodiments, the form is a Type V polymorph. In some embodiments, the form is a Type VI polymorph. In some embodiments, the form is amorphous. Another aspect of the present invention relates to the use of a polymorph or amorphous form of an anhydrate, hydrate, or solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide of the present invention, including any of the foregoing or later embodiments, in the production of a compound for treating or preventing oxidative stress disorders. In some embodiments, the form is a type I polymorph. In some embodiments, the form is a type II polymorph. In some embodiments, the form is a type III polymorph. In some embodiments, the form is a type IV polymorph. In some embodiments, the form is a type V polymorph. In some embodiments, the form is a type VI polymorph. In some embodiments, the form is amorphous.

[0047] For all compositions of the invention, and all methods of using the compositions of the invention, the compositions may include the listed components or steps, or may "essentially include" the listed components or steps. When a composition is described as "essentially including" the listed components, the composition includes the listed components, and may contain other components that do not substantially affect the condition being treated, but does not include any other components that substantially affect the condition being treated, except for those components explicitly listed; or, if the composition does contain additional components other than those components listed that substantially affect the condition being treated, the composition does not contain the additional components in sufficient concentration or amount to substantially affect the condition being treated. When a method is described as "essentially including" the listed steps, the method includes the listed steps, and may contain other steps that do not substantially affect the condition being treated, but does not include any other steps that substantially affect the condition being treated, except for those steps explicitly listed. As a non-limiting specific example, when a composition is described as "essentially including" components, the composition may additionally include any amount of a pharmaceutically acceptable carrier, vehicle, or diluent, and any such component that does not substantially affect the condition being treated.

[0048] Another aspect of the present invention relates to a method for preparing the Type I polymorph of anhydrous form of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the method comprises the steps of: (a) contacting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a liquid containing IPA; and (b) removing the liquid. In some embodiments, step (a) comprises dissolving the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in the liquid. In some embodiments, step (a) comprises slurrying the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in the liquid. In some embodiments, the slurrying in step (a) is carried out for at least about 24 hours. In some embodiments, including any of the preceding embodiments, the liquid is 100% IPA. In some embodiments, including any of the preceding embodiments, the liquid is 98% IPA / 2% water (v / v). In some embodiments, including any of the preceding embodiments, the method further comprises step (a)(i): adding heptane to the liquid. In some embodiments, including any of the preceding embodiments, step (b) comprises filtering the (R)-2 hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. In some embodiments, including any of the preceding embodiments, the mixture in step (a) or step (a)(i) can be seeded with type I crystals. In some embodiments, including any of the preceding embodiments, the purity of the (R)-2 hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in step (a) is at least about 95%. In various embodiments, including any of the foregoing embodiments, the purity of the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in step (a) is at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9%. Another aspect of the present invention relates to an anhydrous form of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide prepared by the above method.

[0049] Another aspect of the present invention relates to a method for preparing a Type II polymorph of an anhydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the method comprises the steps of: (a) dissolving (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in EtOAc, (b) rapidly cooling the mixture obtained from step (a), and (c) isolating the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. In some embodiments, the starting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is Type I. In some embodiments, including any of the preceding embodiments, step (a) is performed at about 60° C. In some embodiments, including any of the preceding embodiments, step (b) comprises rapidly cooling the mixture in an ice bath. In some embodiments, including any of the preceding embodiments, the mixture in step (b) may be seeded with Type II crystals. Another aspect of the present invention relates to an anhydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide prepared by the above method.

[0050] Another aspect of the present invention relates to a method for preparing the III polymorph of the hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the method comprises the steps of: (a) mixing (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and an aqueous solution of 0.5% MC / 2% Tween 80 to produce a slurry; (b) slurrying the mixture obtained in step (a), and (c) removing the aqueous solution of 0.5% MC / 2% Tween 80. "MC" used in the present invention refers to methylcellulose, and "Tween 80" refers to a commercially available polysorbate nonionic surfactant. In some embodiments, the starting (R)-2 hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is Type I. In some embodiments, including any of the preceding embodiments, step (b) is performed for at least about 24 hours. In some embodiments, including any of the preceding embodiments, step (b) is performed at room temperature. In some embodiments, including any of the preceding embodiments, the mixture in step (b) can be seeded with Type III crystals. Another aspect of the present invention relates to a solvate of (R)-2 hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide prepared by the above method.

[0051] Another aspect of the present invention relates to a method for preparing the Type III polymorph of a hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the method comprises the steps of: (a) mixing (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and a 0.5% MC aqueous solution to produce a slurry; (b) slurrying the mixture obtained in step (a), and (c) removing the 0.5% MC aqueous solution. In some embodiments, the starting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is Type I, II, IV, V or VI. In some embodiments, including any of the preceding embodiments, step (b) is performed at room temperature. In some embodiments, including any of the preceding embodiments, step (b) is performed for at least about 7 days. In some embodiments, including any of the preceding embodiments, the mixture in step (b) can be seeded with type III crystals. Another aspect of the present invention relates to a solvate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide prepared by the above process. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 XRPD stacked patterns of short term slurry experiments of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide are shown, a) pattern C from slurrying in 0.5% methylcellulose / 2% Tween 80, b) pattern B from slurrying in tetrahydrofuran (THF), and c) pattern A, starting material.

[0053] Figure 2 Shown are XRPD overlays of amplified samples of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, a) starting material, pattern A, and b) pattern D after subsequent crystallization by evaporation from 2-methyltetrahydrofuran (2-MeTHF).

[0054] Figure 3 XRPD overlays of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide crystallization experiments are shown, a) starting material, pattern A, b) pattern D after subsequent evaporative crystallization from 2-MeTHF (Example 4), c) evaporative crystallization in 2-MeTHF (Example 5, fast cooling), and d) evaporative crystallization in 2-MeTHF (Example 5, slow cooling).

[0055] Figure 4 XRPD stacking patterns of all forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide are shown, a) starting material, pattern A, b) pattern B, c) pattern C, d) pattern D, e) pattern E, rapid cooling crystallization from ethyl acetate (EtOAc), and f) pattern F, rapid cooling crystallization from 2-MeTHF.

[0056] Figure 5 Shown are XRPD stacks of the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide forms, a) Pattern E of rapid cooling crystallization from EtOAc (Example 5), and b) an expanded rapid cooling crystallization from EtOAc (Example 6).

[0057] Figure 6 XRPD stacks of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide forms are shown, a) Pattern B from slurry in THF (Example 3), and b) from a scaled-up slurry in THF (Example 6).

[0058] Figure 7 XRPD stacking patterns of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide forms are shown, a) Pattern C from slurrying in 0.5% methylcellulose / 2% Tween 80 (Example 3), and b) from a scaled-up slurrying in 0.5% methylcellulose / 2% Tween 80 (Example 6).

[0059] Figure 8 Shown are XRPD stacked patterns of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide samples, a) followed by competitive pulping in 0.5% aqueous methylcellulose for 24 hours, b) starting material, pattern A, c) pattern C, and d) followed by competitive pulping in 0.5% aqueous methylcellulose for 7 days.

[0060] Fig. 9Shown are XRPD stacked images of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide samples, a) followed by competitive slurrying in IPA for 24 hours, b) followed by competitive slurrying in IPA for 7 days, c) starting material, mode A, d) followed by competitive slurrying in isopropyl alcohol (IPA) / 2% water for 24 hours, e) competitive slurrying in IPA / 2% water for 7 days.

[0061] Fig.10 The XRPD pattern of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material, Pattern A, is shown.

[0062] Fig.11 An optical microscope image of Pattern A of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material is shown.

[0063] Fig.12 The DSC thermogram of Pattern A of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material is shown.

[0064] Fig.13 Shown is the TGA thermogram of Pattern A of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material.

[0065] Fig.14 The moisture sorption-desorption graph of pattern A of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material is shown.

[0066] Fig.15 Shown are XRPD stacks of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide Pattern E isolated from a single solvent rapid cooling experiment, a) from a 50 mg batch, and b) from a 300 mg scale-up batch.

[0067] Fig.16 An optical microscope image of Pattern E of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0068] Fig.17The DSC thermogram of Pattern E of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0069] Fig.18 The TGA thermogram of Pattern E of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0070] Fig.19 The moisture sorption-desorption graph of Pattern E of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0071] Fig. 20 Shown are XRPD stacks of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide Pattern C slurried in 0.5% methylcellulose / 2% Tween 80, a) from a 50 mg batch, and b) from a 300 mg scaled-up batch.

[0072] Fig.21 An optical microscope image of Pattern C of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0073] Fig. 22 The DSC thermogram of Pattern C of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0074] Fig.23 The TGA thermogram of Pattern C of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0075] Fig.24 The moisture absorption-desorption diagram of Pattern C of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0076] Fig.25 Shown are XRPD stacks of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide Pattern B slurried in THF, a) from a 50 mg batch, and b) from a 300 mg scaled-up batch.

[0077] Fig.26An optical microscope image of Pattern B of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0078] Fig. 27 The DSC thermogram of Pattern B of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0079] Fig.28 The TGA thermogram of Pattern B of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0080] Fig.29 The moisture absorption-desorption diagram of pattern B of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0081] Fig.30 XRPD stacking patterns of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide Pattern D obtained by evaporative crystallization in 2-MeTHF, a) from Example 4, b) Example 5 (fast cooling), c) Example 5 (slow cooling), and d) starting material Pattern A are shown.

[0082] Fig.31 The DSC thermogram of Pattern D of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0083] Fig.32 The TGA thermogram of Pattern D of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0084] Fig.33 XRPD stacking patterns of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide Pattern F obtained by single solvent crystallization in 2-MeTHF, a) from Example 6, b) subsequent moisture sorption analysis (starting from Pattern F obtained in Example 6), and c) Pattern E are shown.

[0085] Fig.34 An optical microscope image of Pattern F of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0086] Fig.35 The DSC thermogram of Pattern F of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0087] Fig.36 The TGA thermogram of Pattern F of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0088] Fig.37 The moisture sorption-desorption diagram of Pattern F of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is shown.

[0089] Fig.38 Shown is a polymorphic form-interaction diagram of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0090] Fig.39 Shown is an XRPD stacking pattern of polymorphic forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0091] Fig.40 The (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material pattern A is shown. 1 H NMR spectroscopy.

[0092] Fig.41 (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide pattern B is shown. 1 H NMR spectroscopy.

[0093] Fig.42 (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide pattern C is shown. 1 H NMR spectroscopy.

[0094] Fig.43 The (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide pattern D is shown. 1 HNMR spectrum.

[0095] Fig.44(R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide pattern E is shown. 1 HNMR spectrum.

[0096] Fig.45 The (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide pattern F is shown. 1 H NMR spectroscopy.

[0097] Detailed description of the invention

[0098] The present invention includes polymorphic and amorphous forms of anhydrates, hydrates, and solvates of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide for treating or inhibiting diseases, developmental delays, and disorders associated with oxidative stress, such as mitochondrial diseases, impaired energy processing disorders, neurodegenerative diseases, and diseases of aging, and methods of using such compositions for treating or inhibiting oxidative stress disorders, or for regulating, normalizing, or enhancing one or more (e.g., 1, 2, 3, or more) energy biomarkers. The present invention further includes methods of producing such polymorphic and amorphous forms.

[0099] Abbreviations used herein have their conventional meanings in the chemical and biological fields unless otherwise indicated.

[0100] The "about" value or parameter mentioned in the present invention includes (and describes) the variation of the value or parameter itself. For example, the description related to "about X" includes the description of "X".

[0101] As used herein, the term "a" or "an" refers to one or more than one, unless the context clearly dictates otherwise.

[0102] "Subject," "individual," or "patient" refers to an individual organism, preferably a vertebrate, more preferably a mammal, and most preferably a human.

[0103] (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide can exist in the form of anhydrate, hydrate, and solvate. Unless otherwise specified or otherwise clearly indicated in the context, the term "(R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide" used in the present invention includes anhydrate, hydrate, and solvate forms of the compound.

[0104] When referring to, for example, an XRPD pattern, a DSC thermogram, or a TGA pattern, the term "substantially as shown" includes patterns that are not necessarily those described herein. Figure 1 The XRPD patterns are consistent, but they all fall within the range of experimental errors or deviations considered by ordinary technicians in the field. For example, in an XRPD pattern, the relative intensities of the peaks in the diffraction pattern can vary, for example, due to sample preparation conditions. In addition, changes in temperature (when generating the XRPD data) can affect the peak position and shape. The XRPD pattern given in the present invention is generated at room temperature (~25°C). In some embodiments, the XRPD pattern is generated at about 15°C to about 30°C. In some embodiments, the XRPD pattern is generated at about 20°C to about 30°C. In some embodiments, the XRPD pattern is generated at about 23°C to about 27°C. In some embodiments, the XRPD pattern is generated at about 24°C to about 26°C. In some embodiments, the XRPD pattern is generated at about 25°C.

[0105] Similarly, when describing a polymorph by characteristic peaks (e.g., the angular position of the peaks), it should be understood that the position of the peaks may vary depending on sample preparation, temperature, etc. The XRPD characteristic peaks given in the present invention are generated at room temperature (~25°C). In some embodiments, the XRPD data are generated at about 15°C to about 30°C. In some embodiments, the XRPD data are generated at about 20°C to about 30°C. In some embodiments, the XRPD data are generated at about 23°C to about 27°C. In some embodiments, the XRPD data are generated at about 24°C to about 26°C. In some embodiments, the XRPD data are generated at about 25°C.

[0106] As used herein, an "isolated" polymorph or amorphous form refers to a form that is at least about 90% of the particular form (i.e., less than 10% of the material contains other forms or other compounds, including but not limited to (S)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide).

[0107] A polymorph composition that is "substantially free" of a particular component means that the composition contains less than about 5% of the particular component. As a non-limiting example, a polymorph composition that is substantially free of Form II polymorph refers to a composition that contains at least about 5% Form II. In some embodiments, "substantially free" means that the composition contains at least about 4%, at least about 3%, at least about 2%, or at least about 1% of the particular component, or wherein the particular component is not within the detection limit.

[0108] "Angular position" refers to the 2θ angle.

[0109] "Treatment" of a disease / condition using the compounds, compositions, and methods of the present invention is defined as administration of one or more compounds or compositions of the present invention, with or without additional therapeutic agents, to reduce or eliminate the disease / condition or one or more symptoms of the disease / condition, or to slow the progression of the disease / condition or one or more symptoms of the disease / condition, or to lessen the severity of the disease / condition or one or more symptoms of the disease / condition. "Inhibition" of a disease / condition using the compounds, compositions, and methods of the present invention is defined as administration of one or more compounds or compositions of the present invention, with or without additional therapeutic agents, to inhibit the clinical manifestation of the disease / condition, or to inhibit the manifestation of adverse symptoms of the disease / condition. The difference between treatment and inhibition is that treatment occurs after the adverse symptoms of the disease / condition manifest in the subject, while inhibition occurs before the adverse symptoms of the disease / condition manifest in the subject. Inhibition can be partial, substantially complete, or complete. Since some diseases / conditions are hereditary, genetic screening can be used to identify patients at risk for the disease / condition. The compounds, compositions, and methods of the present invention can then be applied to asymptomatic patients at risk for developing clinical symptoms of the disease / condition, thereby inhibiting the appearance of any adverse symptoms.

[0110] A "therapeutic use" of the compounds and compositions described herein is defined as the use of one or more compounds or compositions described herein for treating or inhibiting a disease / disorder, as described above. A "therapeutically effective amount" of a compound or composition is an amount of the compound or composition sufficient to modulate, normalize, or enhance one or more energy biomarkers (where modulate, normalize, and enhance are defined below). A "therapeutically effective amount" of a compound or composition is an amount of the compound or composition that, when administered to a subject, is sufficient to reduce or eliminate a disease / disorder or one or more symptoms of a disease / disorder, or to slow the progression of a disease / disorder or one or more symptoms of a disease / disorder, or to reduce the severity of a disease / disorder or one or more symptoms of a disease / disorder, or to inhibit the clinical manifestations of a disease / disorder, or to inhibit the adverse symptomatic manifestations of a disease / disorder. A therapeutically effective amount may be administered in one or more administrations. An "effective amount" of a compound or composition includes a therapeutically effective amount as well as an effective amount that modulates, normalizes, or enhances one or more energy biomarkers of a subject.

[0111] "Modulating" an energy biomarker means changing the level of an energy biomarker to a desired value, or changing the level of an energy biomarker in a desired direction (eg, increase or decrease). Modulation may include, but is not limited to, normalizing and enhancing as defined below.

[0112] "Normalizing" or "normalizing" an energy biomarker is defined as converting the level of the energy biomarker from a pathological value to a normal value, wherein the normal value of the energy biomarker can be 1) the level of the energy biomarker in a healthy person or subject, or 2) the level of the energy biomarker that reduces one or more adverse symptoms in an individual or subject. That is, normalizing an energy biomarker for depression in a disease state means increasing the level of the energy biomarker toward a normal (healthy) value or toward a value that reduces adverse symptoms; normalizing an energy biomarker that is elevated in a disease state means decreasing the level of the energy biomarker toward a normal (healthy) value or toward a value that reduces adverse symptoms.

[0113] "Enhancing" an energy biomarker refers to intentionally altering the level of one or more energy biomarkers away from a normal or pre-enhanced value to achieve a beneficial or desired effect. For example, in situations where a significant energy demand is imposed on a subject, it may be desirable to increase the ATP level in the subject to a level above the normal ATP level for the subject. Enhancing may also be a beneficial effect in subjects with a disease or pathology, such as mitochondrial disease, because normalizing the energy biomarker may not achieve an optimal outcome for the subject; in such situations, enhancing one or more energy biomarkers is beneficial, for example, higher than normal levels of ATP or lower than normal levels of lactate (lactate) are beneficial for such subjects.

[0114] By regulating, normalizing or enhancing the energy biomarker Coenzyme Q, it is meant regulating, normalizing or enhancing the variant of Coenzyme Q that is predominant in the species of interest. For example, the variant of Coenzyme Q that is predominant in humans is Coenzyme Q10. If a species or subject has more than one variant of Coenzyme Q, and the variants are present in large amounts (i.e., when adjusted, normalized or enhanced, present in an amount that can have a beneficial effect on the species or subject), regulating, normalizing or enhancing Coenzyme Q may refer to regulating, normalizing or enhancing any or all variants of Coenzyme Q present in the species or subject.

[0115] "Respiratory chain disorder" refers to a disease / disorder in which the utilization of oxygen by mitochondria, cells, tissues, or individuals is reduced due to defects or disorders in proteins or other components contained in the mitochondrial respiratory chain. "Proteins or other components contained in the mitochondrial respiratory chain" refers to components (including but not limited to proteins, tetrapyrroles, and cytochromes) comprising mitochondrial complexes I, II, III, IV, and / or V. "Respiratory chain proteins" refer to protein components of those complexes, and "respiratory chain protein disorders" refer to diseases / disorders in which the utilization of oxygen by mitochondria, cells, tissues, or individuals is reduced due to defects or disorders in proteins contained in the mitochondrial respiratory chain.

[0116] The term "Parkinson's disease" (also known as "Parkinson's disease" and "Parkinson's syndrome") ("PD") is intended to include not only Parkinson's disease, but also drug-induced Parkinson's syndrome and post-encephalitic Parkinson's syndrome. Parkinson's disease is also known as paralytic palsy or shaking palsy. It is characterized by tremors, muscle rigidity and loss of postural reflexes. The disease usually progresses slowly over a period of 10 to 20 years before symptoms lead to disability. Animals treated with methamphetamine or MPTP have been used to generate Parkinson's disease models because they mimic the effects of Parkinson's disease. These animal models have been used to evaluate the efficacy of various therapies for Parkinson's disease.

[0117] The term "Friedreich's ataxia" is intended to include other related ataxias, sometimes referred to as hereditary ataxias, familial ataxias, or Friedreich's tuberculosis.

[0118] The term "ataxia" is a specific clinical presentation that means a malfunction of the part of the nervous system that coordinates movement, such as the cerebellum. Patients with ataxia have problems with coordination because the parts of the nervous system that control movement and balance are affected. Ataxia may affect the fingers, hands, arms, legs, body, speech, and eye movements. The word ataxia is often used to describe symptoms of incoordination that may be related to infection, injury, other diseases, or degenerative changes in the central nervous system. Ataxia is also used to refer to a group of specific degenerative diseases of the nervous system called hereditary and sporadic ataxias. Ataxia is also often associated with hearing impairment.

[0119] There are three types of ataxia: cerebellar ataxia, including vestibulocerebellar, spinocerebellar, and cerebral cerebellar dysfunction; sensory ataxia; and vestibular ataxia. Examples of diseases classified into spinocerebellar ataxia and multiple system atrophy are hereditary olivopontocerebellar atrophy, hereditary cerebellar cortical atrophy, Friedrich's ataxia, Machado-Joseph disease, Ramsay Hunt syndrome, hereditary dentatorubral-pallidoluysian atrophy, hereditary spastic paraplegia, Shy-Drager syndrome, corticocerebellar atrophy, striatonigral degeneration, Marinesco-Sjogren syndrome, alcoholic corticocerebellar atrophy, paraneoplastic cerebellar atrophy associated with malignant tumors, toxic cerebellar atrophy caused by toxic substances, vitamin E deficiency due to tocopherol transport protein (aTTP) mutation or lipid absorption disorder such as abetalipoproteinemia, cerebellar atrophy associated with endocrine disorders, etc.

[0120] Examples of ataxia symptoms include motor ataxia, truncal ataxia, limb ataxia, etc., autonomic nervous system disorders such as orthostatic hypotension, dysuria, oligohidrosis, sleep apnea, orthostatic syncope, etc., lower limb rigidity, nystagmus, oculomotor nerve disorders, pyramidal tract dysfunction, extrapyramidal symptoms (postural adjustment dysfunction, muscle rigidity, akinesia, tremor), dysphagia, tongue atrophy, posterior cord symptoms, muscle atrophy, muscle weakness, deep hyperreflexia, sensory disorders, scoliosis, kyphoscoliosis, foot deformities, dysarthria, dementia, manic state, decreased motivation for rehabilitation, etc.

[0121] Polymorphic and amorphous forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0122] The present invention provides various crystalline and amorphous forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide:

[0123]

[0124] (R)-2-Hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dien-1-yl)butanamide

[0125] and methods of producing such forms, and methods of using such forms.

[0126] Table 1 below provides a summary of certain polymorphic forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide according to the present invention.

[0127] Table 1: Summary of characterization of polymorphic forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0128]

[0129]

[0130] Fig.38 A diagram of the interactions between the various polymorphic forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide is provided. Fig.39 Shown is an XRPD stacking pattern of polymorphic forms of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0131] Table A provides various embodiments of the angular positions of certain characteristic peaks of X-ray powder diffraction of the polymorphic forms of the present invention. In some embodiments, the angular position may vary by ±0.2. In some embodiments, the angular position may vary by ±0.1. In some embodiments, the angular position may vary by ±0.05. In some embodiments, the angular position may vary by ±0.02. Table AI-VI Angular Positions of Certain Characteristic Peaks of X-ray Powder Diffraction

[0132]

[0133]

[0134]

[0135]

[0136] Tables 2-7 provide additional embodiments of the angular positions of the characteristic peaks of the X-ray powder diffraction of the polymorphic forms of the present invention. In some embodiments, the polymorphic forms are characterized by the angular positions of the characteristic peaks shown in Table A. In some embodiments, the polymorphic forms are characterized by 3 or more (e.g., 4, 5, 6, 7, 8, 9, 10, or more than 10) angular positions of the characteristic peaks of the X-ray powder diffraction shown in the following Tables 2-7. In some embodiments, the angular position may vary by ±0.2. In some embodiments, the angular position may vary by ±0.1. In some embodiments, the angular position may vary by ±0.05. In some embodiments, the angular position may vary by ±0.02.

[0137] As non-limiting examples, the Form I polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions shown in Table 2.

[0138] Table 2. Angular positions of characteristic peaks of X-ray powder diffraction of mode A (anhydrous form, type I)

[0139] The Form I polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0140]

[0141]

[0142] In some embodiments, the Form I polymorph is characterized by at least 3 or more angular positions. In some embodiments, the angular positions include 12.06, 17.03, and 17.26 ± 0.2. In some embodiments, the Form I polymorph is characterized by at least 4 or more angular positions. In some embodiments, the at least 4 angular positions include 12.1, 17.0, 17.3, and 15.33 ± 0.2. In some embodiments, the at least 4 angular positions include 12.1, 17.0, 17.3, 15.33, and 18.72 ± 0.2.

[0143] Table 3. Angular positions of characteristic peaks of X-ray powder diffraction of mode D (2-MeTHF solvate, V form)

[0144] The Form V polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0145]

[0146]

[0147] Table 4. Angular positions of characteristic peaks of X-ray powder diffraction of mode C (hydrate, form III)

[0148] The Form III polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0149]

[0150] Table 5. Angular positions of characteristic peaks of X-ray powder diffraction of mode E (anhydrous form, type II)

[0151] The Form II polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0152]

[0153]

[0154] Table 6. Angular positions of characteristic peaks of X-ray powder diffraction of mode B (THF solvate, Form IV)

[0155] The Form IV polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0156]

[0157] Table 7. Angular positions of characteristic peaks of X-ray powder diffraction of pattern F (2-MeTHF solvate, Form VI)

[0158] The Form VI polymorph may be characterized by 3, 4, 5, 6, 7, 8, 9, 10, or more angular positions as shown below:

[0159] 2θ angle Spacing d, A 2θ angle Spacing d, A 4.39 20.1 16.91 5.2 6.27 14.1 17.33 5.1 7.00 12.6 17.59 5.0 8.62 10.3 18.33 4.8 8.85 10.0 18.75 4.7 9.41 9.4 19.13 4.6 9.91 8.9 20.25 4.4 11.32 7.8 20.76 4.3 11.50 7.7 21.68 4.1 12.25 7.2 22.06 4.0 12.56 7.0 22.27 4.0 12.94 6.8 22.61 3.9 13.29 6.7 22.94 3.9 14.03 6.3 24.01 3.7 14.82 6.0 24.33 3.7 15.10 5.9 24.65 3.6 15.44 5.7 25.48 3.5 15.71 5.6 26.05 3.4 16.01 5.5 28.63 3.1 16.67 5.3 29.18 3.1

[0160] Some polymorphic or amorphous forms of a drug may have superior characteristics compared to other forms that affect the desirability of the drug from a pharmaceutical and / or manufacturing perspective, such as: improved stability, increased solubility, better handling properties, lack of associated toxic solvents (e.g., solvates of toxic solvents), improved purity, better particle size and / or particle size distribution, improved bulk density, and ease of manufacture.

[0161] Forms I, II, III and the amorphous form are anhydrates or hydrates and advantageously are not solvates with toxic solvents (eg, THF and 2-MeTHF).

[0162] Forms I-IV and VI exhibit good water solubility (>1.3 mg / ml for each form), with Form I having the highest water solubility at 1.74 mg / ml (Example 10). In addition, Form I was shown to be soluble in a variety of polar and non-polar solvents (Example 2), indicating the ability to be administered using a variety of solvents. In some embodiments, drugs with a physiological log D close to zero are advantageous; thus solubility in polar and non-polar solvents indicates a more favorable physiological log D. Form I further exhibits increased solubility in a simple detergent (0.5% MC / 2% Tween 80) (Example 2); this solubility in a simple detergent may be advantageous because these conditions can simulate intestinal conditions for oral administration of drugs. Form III (hydrate) is formed under simple detergent conditions, and thus Form III may be the form of the drug that will be produced in the intestine.

[0163] Forms I, II and III advantageously demonstrate stability to elevated humidity (Example 8). Form I can also be tested by grinding and shows stability to grinding (Example 9). As shown in the Examples, the experiments performed show that Form I is highly stable.

[0164] Non-hygroscopic forms are easier to handle from a manufacturing standpoint; as shown in Example 11, Forms I, II, IV and VI are non-hygroscopic.

[0165] Certain particle shapes and sizes may be advantageous: particles that are closer to spherical may be preferred, with flakes and needles being less preferred. As shown in Example 11, Types I, II, and III have more advantageous shapes, while Type VI is flake-shaped and Type IV is needle-shaped. With regard to particle size, smaller, more uniform sizes may be preferred. Smaller particles may have increased bioavailability, may be more easily dissolved, and may be easier to handle due to the reduction in drying time. In addition, smaller particles may not require the micronization step that may be required for larger particles. As shown, Types I-III have more advantageous particle sizes than Types IV and VI.

[0166] A higher melting point may indicate a form with improved handling characteristics (e.g., easier drying and processing) and greater thermal stability. Form I has the highest melting point of Forms I-VI. Additionally, in some embodiments, a single peak may be preferred, as multiple peaks may indicate conversion to different forms. Form I has a single DSC peak, and all other forms have 2 or 3 peaks.

[0167] Various forms (polymorphs and amorphous forms) of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide can also be used as intermediates in the preparation of the desired form. As a non-limiting example, if the preferred synthetic method for preparing (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide results in the production of a non-preferred form, the non-preferred form can be used as an intermediate in the preparation of the desired form.

[0168] Diseases suitable for treatment or inhibition by the compositions and methods of the present invention

[0169] A variety of diseases / disorders are believed to be caused or exacerbated by oxidative stress that affects normal electron flow in cells, such as mitochondrial diseases, impaired energy processing disorders, neurodegenerative diseases, and diseases of aging, and diseases that can be treated and inhibited using the polymorphic and amorphous forms and methods of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide described herein.

[0170] Non-limiting examples of oxidative stress disorders include, for example, mitochondrial diseases (including inherited mitochondrial diseases) such as Alpers Disease, Barth syndrome, beta-oxidation defects, carnitine-acyl-carnitine deficiency, carnitine deficiency, creatine deficiency, coenzyme Q10 deficiency, complex I deficiency, complex II deficiency, complex III deficiency, complex IV deficiency, complex V deficiency, COX deficiency, chronic progressive external ophthalmoplegia (CPEO), CPT I deficiency; CPT II deficiency, Friedreich's ataxia (FA), glutaric aciduria type II, Kearns-Sayre syndrome (KSS), lactic acidosis, long chain acyl-CoA dehydrogenase deficiency (LCAD), LCHAD, Leigh Disease or Syndrome (subacute necrotizing encephalopathy, Leigh Disease or Syndrome, Leigh-like syndrome, Leber's hereditary optic neuropathy (LHON, also known as Leber's disease, Leber's optic atrophy (LOA), or Leber's optic neuropathy (LON)), fatal infantile cardiomyopathy (LIC), Luft disease, multiple acyl-CoA dehydrogenase deficiency (MAD), medium-chain acyl-CoA dehydrogenase deficiency (MCAD), mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), myoclonic seizures with ragged red fibers (MERRF), mitochondrial recessive ataxia Mitochondrial dysregulation syndrome (MIRAS), mitochondrial cell disease, mitochondrial DNA depletion, mitochondrial encephalopathy, mitochondrial myopathy, mitochondrial neurogastrointestinal encephalopathy (MNGIE), neuropathy, ataxia, and retinitis pigmentosa (NARP), Pearson syndrome, pyruvate carboxylase deficiency, pyruvate dehydrogenase deficiency, POLG mutation, respiratory chain disorders, short-chain acyl-CoA dehydrogenase deficiency (SCAD), SCHAD, very long-chain acyl-CoA dehydrogenase deficiency (VLCAD); myopathies such as cardiomyopathy and encephalomyopathy; neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig's disease (Lou Gehrig's disease ( Gehrig's disease); motor neuron disease; neurological diseases such as epilepsy; age-related diseases, especially diseases for which Coenzyme Q10 has been proposed for treatment, such as macular degeneration, diabetes (e.g., type 2 diabetes), metabolic syndrome, and cancer (e.g., brain cancer); genetic diseases such as Huntington's disease (which is also a neurological disease); mood disorders such as schizophrenia and bipolar disorder;Pervasive developmental disorders such as autism, Asperger's syndrome, childhood disintegrative disorder (CDD), Rett's disorder, and PDD-NOS; cerebrovascular accidents such as stroke; visual disorders such as those caused by neurodegenerative diseases of the eye such as optic neuropathy, Leber's hereditary optic neuropathy, dominantly inherited juvenile optic atrophy, optic neuropathy caused by toxic drugs, glaucoma, age-related macular degeneration ("dry" or non-exudative macular degeneration and "wet" or exudative macular degeneration), Stargardt's macular dystrophy ( dystrophy, diabetic retinopathy, diabetic maculopathy, retinopathy of prematurity, or retinal damage associated with ischemia-reperfusion; diseases caused by energetic damage including those due to oxygen deprivation, oxygen toxicity, or toxic oxygen, and qualitative or quantitative disruptions in oxygen transport such as hemoglobinopathies, for example, thalassemia or sickle cell anemia; other diseases caused by mitochondrial dysfunction such as neuronal damage from excitotoxic agents, such as those associated with epilepsy, stroke, and ischemia; and other diseases including renal tubular acidosis; attention-deficit / hyperactivity disorder (ADHD); neurodegenerative diseases resulting in hearing or balance impairment; dominant optic atrophy (DOA); maternally inherited diabetes and deafness (MIDD); chronic fatigue; contrast-induced renal damage; contrast-induced retinopathy damage; abetalipoproteinemia; retinal atrophy; Wolfram disease; Tourette syndrome syndrome); cobalamin C deficiency; methylmalonic aciduria; glioblastoma; Down syndrome; acute tubular necrosis; muscular dystrophy; leukodystrophy; progressive supranuclear palsy; spinal muscular atrophy; hearing loss (e.g., noise-induced hearing loss); traumatic brain injury; juvenile Huntington's disease; multiple sclerosis; NGLY1; multiple system atrophy; adrenoleukodystrophy; and adrenomyeloneuropathy. It should be understood that certain specific diseases or conditions may fall into more than one category; for example, Huntington's disease is a genetic disease as well as a disease of the nervous system. In addition, certain oxidative stress diseases and conditions may also be considered mitochondrial diseases. ;

[0171] For some diseases suitable for treatment with the compounds and methods of the present invention, the main cause of the disease is due to defects in the respiratory chain or another defect that prevents the normal utilization of energy in mitochondria, cells or tissues. Non-limiting examples of diseases falling into this category include hereditary mitochondrial diseases, such as myoclonic seizures with broken red fibers (MERRF), mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like attacks (MELAS), Leber's hereditary optic neuropathy (LHON, also known as Leber's disease, Leber's optic atrophy (LOA), or Leber's optic neuropathy (LON)), Leigh disease or Leigh syndrome, Kearns-Sayre syndrome (KSS), and Friedreich's ataxia (FA). For some diseases suitable for treatment with the compounds and methods of the present invention, the main cause of the disease is not due to defects in the respiratory chain or other defects that prevent the normal utilization of energy in mitochondria, cells or tissues; non-limiting examples of diseases falling into this category include stroke, cancer, and diabetes. However, these latter diseases are particularly aggravated by energy damage and are particularly suitable for treatment with the compounds of the present invention to improve the condition. Relevant examples of such diseases include ischemic stroke and hemorrhagic stroke, where the main cause of the disease is impaired blood supply to the brain. Although ischemic attacks caused by thrombosis or embolism, or hemorrhagic attacks caused by ruptured blood vessels are not primarily caused by defects in the respiratory chain or another metabolic defect that prevents normal utilization of energy, oxidative stress plays a role in the ischemic cascade caused by oxygen reperfusion injury caused by hypoxia (such cascade occurs in heart attacks and strokes). Therefore, treatment with the compounds and methods of the present invention will reduce the effects of the disease, disorder or condition. Modulating one or more energy biomarkers, normalizing one or more energy biomarkers, or enhancing one or more energy biomarkers may also prove to be beneficial as a therapeutic measure and preventive measure in such diseases. For example, for patients who are scheduled for non-urgent repair of aneurysms, if the aneurysm ruptures before successful repair, energy biomarkers should be enhanced before and during surgery to improve the patient's prognosis.

[0172] The term "oxidative stress disorder" or "oxidative stress disease" encompasses diseases caused by oxidative stress and diseases aggravated by oxidative stress. The term "oxidative stress disorder" or "oxidative stress disease" encompasses diseases and conditions whose primary cause is due to a defect in the respiratory chain or another defect that prevents the normal utilization of energy in mitochondria, cells or tissues, and diseases and conditions whose primary cause is not due to a defect in the respiratory chain or another defect that prevents the normal utilization of energy in mitochondria, cells or tissues. The former group of diseases may be referred to as "primary oxidative stress disorders" and the latter as "secondary oxidative stress disorders". It should be noted that the distinction between "diseases caused by oxidative stress" and "diseases aggravated by oxidative stress" is not absolute; a disease may be both a disease caused by oxidative stress and a disease aggravated by oxidative stress. The boundary between "primary oxidative stress disorder" and "secondary oxidative stress disorder" is more distinct as long as there is only one primary cause of the disease or condition and the primary cause is known.

[0173] Given the somewhat fluid border between diseases caused by oxidative stress and diseases exacerbated by oxidative stress, mitochondrial diseases or disorders and impaired energy processing diseases and disorders tend to fall into the category of diseases caused by oxidative stress, while neurodegenerative diseases and diseases of aging tend to fall into the category of diseases exacerbated by oxidative stress. Mitochondrial diseases or disorders and impaired energy processing diseases and disorders are often primary oxidative stress disorders, while neurodegenerative diseases and diseases of aging may be either primary or secondary oxidative stress disorders.

[0174] Clinical evaluation and treatment effects of oxidative stress

[0175] Several easily measured clinical markers are used to assess the metabolic status of patients with oxidative stress disorders. These markers can also be used as indicators of the effectiveness of a given treatment, as the levels of the markers are converted from pathological values ​​to healthy values. Such clinical markers include, but are not limited to, energy biomarkers such as lactate (lactate) levels in whole blood, plasma, cerebrospinal fluid or ventricular fluid; pyruvate (pyruvate) levels in whole blood, plasma, cerebrospinal fluid or ventricular fluid; lactate (root) / pyruvate (root) ratio in whole blood, plasma, cerebrospinal fluid or ventricular fluid; total, reduced or oxidized glutathione levels, or reduced / oxidized glutathione ratios in whole blood, plasma, lymphocytes, cerebrospinal fluid or ventricular fluid; total, reduced or oxidized cysteine ​​levels, or reduced / oxidized cysteine ​​ratios in whole blood, plasma, lymphocytes, cerebrospinal fluid or ventricular fluid; creatine phosphate levels, NADH (NADH+H+) or NADPH (NADPH+H+) levels; NAD or NADP levels; ATP levels; anaerobic threshold; reduced coenzyme Q (CoQ 还原 ) level; oxidized coenzyme Q (CoQ 氧化 ) levels; total coenzyme Q (CoQ总 ) levels; oxidized cytochrome C levels; reduced cytochrome C levels; oxidized cytochrome C / reduced cytochrome C ratio; acetoacetate / root level, β-hydroxybutyrate / root level, acetoacetate / β-hydroxybutyrate ratio, 8-hydroxy-2'-deoxyguanosine (8-OHdG) level; reactive oxygen species levels; and oxygen consumption (VO2) levels, carbon dioxide output (VCO2) levels, and respiratory quotient (VCO2 / VO2). Several of these clinical indices are routinely measured in exercise physiology laboratories and provide a convenient assessment of the metabolic state of a subject. In one embodiment of the invention, the level of one or more energy biomarkers in a patient with an oxidative stress disorder such as Friedreich's ataxia (FA), Leber's hereditary optic neuropathy, MELAS, KSS, or CoQ10 (coenzyme Q10) deficiency is improved to within two standard deviations of the mean level of healthy subjects. In another embodiment of the invention, the levels of one or more energy biomarkers in patients with oxidative stress disorders, such as Friedreich's ataxia (FA), Leber's hereditary optic neuropathy, MELAS, KSS, or CoQ10 deficiency, are improved to within one standard deviation of the mean level of healthy subjects. Exercise intolerance can also be used as an indicator of the efficacy of a given treatment, wherein an improvement in exercise tolerance (i.e., a reduction in exercise intolerance) indicates the efficacy of a given treatment.

[0176] Several metabolic biomarkers have been used to evaluate the efficacy of CoQ10, and these metabolic biomarkers can be monitored as energy biomarkers for the methods of the present invention. Lactate / root is an anaerobic metabolite of glucose, which is removed by reduction to pyruvate / root in an aerobic environment or by oxidative metabolism, which depends on a functional mitochondrial respiratory chain. Dysfunction of the respiratory chain can lead to insufficient removal of lactate / root and pyruvate / root from the circulation, and elevated lactate (root) / pyruvate (root) ratios are observed in mitochondrial cell pathology (see Scriver CR, The metabolic and molecular bases of inherited disease, 7th ed., New York: McGraw-Hill, Health Professions Division, 1995; and Munnich et al., J. Inherit. Metab. Dis. 15 (4): 448-55 (1992)). Therefore, the blood lactate / pyruvate ratio (Chariot et al., Arch. Pathol. Lab. Med. 118(7):695-7 (1994)) is widely used as a noninvasive test for mitochondrial cytopathies (see again Scriver CR, The metabolic and molecular bases of inherited disease, 7th ed., New York: McGraw-Hill, Health Professions Division, 1995; and Munnich et al., J. Inherit. Metab. Dis. 15(4):448-55 (1992)) and toxic mitochondrial myopathy (Chariot et al., Arthritis Rheum. 37(4):583-6 (1994)). Changes in the redox state of liver mitochondria can be studied by measuring the arterial ketone body ratio (acetoacetate / 3-hydroxybutyrate: AKBR) (Ueda et al., J. Cardiol. 29(2):95-102 (1997)).Urinary excretion of 8-hydroxy-2'-deoxyguanosine (8-OHdG) is commonly used as a biomarker to assess the extent of repair of ROS-induced DNA damage in clinical and occupational settings (Erhola et al., FEBS Lett. 409(2):287-91 (1997); Honda et al., Leuk. Res. 24(6):461-8 (2000); Pilger et al., Free Radic. Res. 35(3):273-80 (2001); Kim et al. Environ Health Perspect 112(6):666-71 (2004)).

[0177] Magnetic resonance spectroscopy (MRS) has been used to diagnose mitochondrial cell disease by using proton MRS (1H-MRS) to show elevated lactate / radical in cerebrospinal fluid (CSF) and cortical white matter (Kaufmann et al., Neurology 62(8):1297-302 (2004)). Phosphorus MRS (31P-MRS) has been used to show low levels of cortical phosphocreatine (PCr) (Matthews et al., Ann. Neurol. 29(4):435-8 (1991)), and delayed PCr recovery kinetics after exercise in skeletal muscle (Matthews et al., Ann. Neurol. 29(4):435-8 (1991); Barbiroli et al., J. Neurol. 242(7):472-7 (1995); Fabrizi et al., J. Neurol. Sci. 137(1):20-7 (1996)). Low skeletal muscle PCr has been demonstrated by direct biochemical assays in patients with mitochondrial cytopathies.

[0178] Exercise testing is particularly useful as an assessment and screening tool for mitochondrial myopathies. One of the hallmark features of mitochondrial myopathies is a decrease in maximal whole-body oxygen consumption (VO2 最大 ) (Taivassalo et al., Brain 126(Pt2):413-23(2003)). 最大Some mitochondrial myopathies affect cardiac function, determined by cardiac output (Qc) and peripheral oxygen extraction (arterial-venous total oxygen content) differences, in which delivery can be altered; however, most mitochondrial myopathies show characteristic defects in peripheral oxygen extraction (AV O2 difference) and enhanced oxygen delivery (hyperdynamic circulation) (Taivassalo et al., Brain 126 (Pt 2): 413-23 (2003)). This can be demonstrated by the lack of exercise-induced venous deoxygenation, measured by direct AV balance (Taivassalo et al., Ann. Neurol. 51 (1): 38-44 (2002)) and non-invasive near-infrared spectroscopy (Lynch et al., Muscle Nerve 25 (5): 664-73 (2002); van Beekvelt et al., Ann. Neurol. 46 (4): 667-70 (1999)).

[0179] Several of these energy biomarkers are discussed in more detail below. It should be emphasized that although certain energy biomarkers are discussed and listed in the present invention, the present invention is not limited to the modulation, normalization or enhancement of only these listed energy biomarkers.

[0180] Lactate (lactate) levels: Mitochondrial dysfunction often results in abnormal lactate levels, elevated pyruvate levels, and conversion of pyruvate to lactate to maintain glycolytic capacity. Mitochondrial dysfunction can also result in abnormal NADH+H+, NADPH+H+, NAD, or NADP levels because reduced nicotinamide adenine dinucleotide cannot be efficiently processed by the respiratory chain. Lactate levels can be measured by collecting samples of appropriate body fluids such as whole blood, plasma, or cerebrospinal fluid. Using magnetic resonance, lactate levels can be measured in almost any volume required in the body, such as required by the brain.

[0181] The use of magnetic resonance imaging to measure brain lactic acidosis in MELAS patients is documented in Kaufmann et al., Neurology 62(8):1297(2004). Intraventricular lactate levels in the brain are associated with two mutations that cause MELAS, A3243G and A8344G. Whole blood, plasma, and cerebrospinal fluid lactate levels can be measured by commercially available equipment such as the YSI 2300STATPlus Glucose & Lactate Analyzer (YSI Life Sciences, Ohio).

[0182] NAD, NADP, NADH and NADPH levels: NAD, NADP, NADH (NADH+H+) or NADPH (NADPH+H+) can be measured by various fluorescent, enzymatic or electrochemical techniques, such as electrochemical analysis described in US2005 / 0067303.

[0183] GSH, GSSG, Cys, and CySS levels: Briefly, plasma levels of GSH, GSSG, Cys, and CySS were used to calculate the in vivo E h Value. Samples were collected using the procedure reported by Jones et al. (2009 Free Radical Biology & Medicine 47 (10) pp1329-1338), and ethyl bromide was used to alkylate free thiols, and HPLC and electrochemical or MSMS were used to separate, detect and quantify the molecules. More details are described in PCT application number PCT / US2013 / 058568, and methods for different experimental parameters were developed to analyze the most common monothiols and disulfides (cystine, cysteine, reduced (GSH) and oxidized glutathione (GSSG)) present in human plasma, and bathophenanthroline disulfonic acid was used as an internal standard (Is). All target analytes and IS were completely separated at 35 °C on a C18 RP column (250 mm × 4.6 mm, 3 μm) using 0.2% TFA:acetonitrile as the mobile phase at a pump delivery rate of 0.6 ml / min and a detection potential of 1475 mV in direct current (DC) mode using an electrochemical detector.

[0184] Oxygen consumption (vO2 or VO2), carbon dioxide output (vCO2 or VCO2), and respiratory quotient (VCO2 / VO2): vO2 is usually higher at rest (resting vO2) or at maximal exercise intensity (vO2 最大 Optimally, both values ​​would be measured. However, in severely disabled patients, vO2 最大 The measurement of vO2 may be impractical. Both forms of vO2 measurement are readily accomplished using standard equipment from various suppliers, such as Korr Medical Technologies, Inc. (Salt Lake City, Utah). VCO2 can also be easily measured, and the ratio of VCO2 to VO2 under the same conditions (VCO2 / VO2, resting or maximum exercise intensity) provides the respiratory quotient (RQ).

[0185] Oxidized cytochrome C, reduced cytochrome C, and the ratio of oxidized cytochrome C to reduced cytochrome C: Cytochrome C parameters, such as oxidized cytochrome C levels (Cyt C 氧化 ), reduced cytochrome C level (Cyt C还原 ), and the ratio of oxidized cytochrome C / reduced cytochrome C (Cyt C 氧化 ) / (Cyt C 还原 ) can be measured by in vivo near-infrared spectroscopy. See, for example, Rolfe, P., "In vivo near-infrared spectroscopy," Annu. Rev. Biomed. Eng. 2:715-54 (2000) and Strangman et al., "Non-invasive neuroimaging using near-infrared light" Biol. Psychiatry 52:679-93 (2002).

[0186] Exercise tolerance / exercise intolerance: Exercise intolerance is defined as "a decrease in the ability to perform dynamic movements involving large skeletal muscles due to symptoms of dyspnea or fatigue" ( et al., Circulation 107:1210 (2003)). Exercise intolerance is often accompanied by myoglobinuria due to the destruction of muscle tissue and the subsequent excretion of myoglobin in the urine. Various measures of exercise intolerance can be used, such as time spent walking or running on a treadmill before exhaustion, time spent on a bicycle exercise (stationary bicycle) before exhaustion, etc. Treatment with the compounds, compositions or methods of the invention can result in an increase in exercise tolerance of about 10% or more (e.g., an increase in time to exhaustion of about 10% or more, e.g., from 10 minutes to 11 minutes), an increase in exercise tolerance of about 20% or more, an increase in exercise tolerance of about 30% or more, an increase in exercise tolerance of about 40% or more, an increase in exercise tolerance of about 50% or more, an increase in exercise tolerance of about 75% or more, or an increase in exercise tolerance of about 100% or more. While exercise tolerance is not strictly speaking an energy biomarker, for purposes of the present invention, modulating, normalizing or enhancing an energy biomarker includes modulating, normalizing or enhancing exercise tolerance.

[0187] Similarly, for pyruvate levels, lactate / pyruvate ratio, APT levels, anaerobic threshold, reduced coenzyme Q (CoQ 还原 ) levels, oxidized coenzyme Q (CoQ 氧化 ) level, total coenzyme Q (CoQ 总Tests for normal and abnormal values ​​of ) levels, oxidized cytochrome C levels, reduced cytochrome C levels, oxidized cytochrome C / reduced cytochrome C ratios, GSH and cysteine ​​reduced, oxidized, total levels and ratios, acetoacetate / root levels, β-hydroxybutyrate / root levels, acetoacetate (root) / β-hydroxybutyrate (root) ratios, 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, and reactive oxygen species levels are known in the art and can be used to evaluate the efficacy of the compounds, compositions, and methods of the invention. (For purposes of the invention, modulating, normalizing, or enhancing an energy biomarker includes modulating, normalizing, or enhancing the anaerobic threshold.)

[0188] Table 8 below illustrates the effects of various disorders on biochemical and energy biomarkers. It also indicates the physical effects (such as disease symptoms or other effects of the disorder) that are typically associated with a given disorder. It should be noted that any of the energy biomarkers listed in the table, in addition to the energy biomarkers listed elsewhere herein, may also be modulated, enhanced or normalized by the compounds, compositions and methods of the present invention. RQ = respiratory quotient; BMR = basal metabolic rate; HR (CO) = heart rate (cardiac output); T = body temperature (preferably measured as core temperature); AT = anaerobic threshold; pH = blood pH (venous and / or arterial).

[0189] Table 8

[0190]

[0191] Treatment of a subject afflicted with an oxidative stress disorder according to the methods of the present invention may result in inducing a reduction or remission of symptoms in the subject, such as halting further progression of the disease.

[0192] Partial or complete inhibition of oxidative stress disorders may result in a reduction in the severity of one or more symptoms otherwise experienced by the subject. For example, partial inhibition of MELAS may result in a reduction in the number of strokes or epileptic seizures.

[0193] Any one or any combination of the energy biomarkers described herein provides a convenient measurable baseline by which to determine the effectiveness of a treatment or inhibitory therapy. In addition, other energy biomarkers are known to those skilled in the art and can be monitored to assess the efficacy of a treatment or inhibitory therapy.

[0194] Use of compounds or compositions to modulate energy biomarkers

[0195] In addition to monitoring energy biomarkers to assess the status of treating or inhibiting oxidative stress diseases, the compounds or compositions of the present invention can be used in subjects or patients to modulate one or more energy biomarkers. Modulating energy biomarkers can be accomplished to normalize energy biomarkers in a subject, or to enhance energy biomarkers in a subject.

[0196] Normalizing one or more energy biomarkers is defined as restoring the level of one or more such energy biomarkers to normal or near normal levels for a subject whose level of one or more energy biomarkers exhibits a pathological difference from normal levels (i.e., levels in healthy subjects), or altering the level of one or more energy biomarkers to alleviate pathological symptoms in a subject. Depending on the nature of the energy biomarkers, these levels may exhibit measured values ​​that are higher or lower than normal values. For example, pathological lactate levels are generally higher than lactate levels in normal (i.e., healthy) people, and a decrease in such levels may be desirable. Pathological ATP levels are generally lower than ATP levels in normal (i.e., healthy) people, and an increase in ATP levels may be desirable. Thus, normalizing an energy biomarker may involve restoring the level of the energy biomarker to within about at least two standard deviations of the normal value for the subject, more preferably to within about at least one standard deviation of the normal value for the subject, to within about at least one half of the normal value for the subject, or to within about at least one quarter of the normal value.

[0197] Enhancing the level of one or more energy biomarkers is defined as changing the level of one or more energy biomarkers present in a subject to a level that provides a beneficial or desired effect to the subject. For example, a person who undergoes strenuous or prolonged physical activity, such as mountain climbing, may benefit from increased ATP levels or decreased lactate levels. As described above, normalizing energy biomarkers may not be optimal for subjects with oxidative stress diseases, and such subjects may also benefit from enhancing energy biomarkers. Examples of subjects who may benefit from enhanced levels of one or more energy biomarkers include, but are not limited to, subjects who undergo strenuous or prolonged physical activity, subjects with chronic energy problems, or subjects with chronic breathing problems. Such subjects include, but are not limited to, pregnant women, especially those in labor; newborns, especially premature newborns; subjects exposed to extreme environments, such as heat (normal temperatures exceeding about 85-86 degrees Fahrenheit or about 30 degrees Celsius for about 4 hours or more per day), cold (normal temperatures below about 32 degrees Fahrenheit or about 0 degrees Celsius for about 4 hours or more per day), or below average oxygen levels, above average carbon dioxide levels, or above average atmospheric pollution (airline travelers, flight personnel, subjects at high altitudes, people living in cities with below average air quality levels, subjects working in closed environments with deteriorating air quality); subjects with lung disease or below average lung capacity, such as patients with tuberculosis, lung cancer, emphysema, and cystic fibrosis; subjects recovering from surgery or illness; elderly subjects, including elderly subjects experiencing energy decline; subjects with chronic fatigue, including chronic fatigue syndrome; subjects experiencing acute trauma; subjects in shock; subjects requiring acute oxygen therapy; subjects requiring chronic oxygen therapy; or other subjects with acute, chronic, or sustained energy needs that would benefit from enhancement of energy biomarkers.

[0198] Thus, when an increase in the level of one or more energy biomarkers is beneficial to a subject, enhancement of the one or more energy biomarkers may involve increasing the corresponding energy biomarker or energy biomarker level to more than about one quarter standard deviation of normal, to more than about one half standard deviation of normal, to more than about at least one standard deviation of normal, or to more than about at least two standard deviations of normal. Alternatively, the level of the one or more energy biomarkers may be increased by about at least 10% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 20% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 30% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 40% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 50% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 75% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement, or by about at least 100% above the subject's level of the corresponding one or more energy biomarkers prior to enhancement.

[0199] When it is desired to reduce the level of one or more energy biomarkers in order to enhance one or more energy biomarkers, the level of the one or more energy biomarkers may be reduced by an amount of about at least one quarter standard deviation of normal value in a subject, by an amount of about at least one half standard deviation of normal value in a subject, by an amount of about at least one standard deviation of normal value in a subject, or by an amount of about two standard deviations of normal value in a subject. Alternatively, the level of the one or more energy biomarkers may be reduced by about at least 10% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 20% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 30% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 40% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 50% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, by about at least 75% below the subject level of the corresponding one or more energy biomarkers prior to enhancement, or by about at least 90% below the subject level of the corresponding one or more energy biomarkers prior to enhancement.

[0200] Use of compounds or compositions in research applications, experimental systems, and analytical tests

[0201] The compounds or compositions of the present invention may also be used in research applications. They may be used in vitro, in vivo or ex vivo experiments to modulate one or more energy biomarkers in an experimental system. Such experimental systems may be cell samples, tissue samples, cell components or mixtures of cell components, partial organs, whole organs, or organisms. Any one or more of the compounds or compositions may be used in experimental systems or research applications. Such research applications may include, but are not limited to, use as assay reagents in the presence / absence of one or more compounds or compositions of the present invention, elucidation of biochemical pathways, or evaluation of the effects of other drugs on the metabolic state of an experimental system.

[0202] In addition, the compounds or compositions of the present invention can be used for biochemical tests or detections. Such tests may include incubating one or more compounds or compositions of the present invention with tissue or cell samples from a subject to evaluate the potential response of the subject (or the response of a specific subtype of the subject) to administer the one or more compounds or compositions, or determine which compound or composition of the present invention produces the best effect in a specific category or subset of the subject. One such test or detection will involve 1) obtaining a cell sample or tissue sample from a subject, in which the regulation of one or more energy biomarkers can be determined; 2) administering one or more compounds or compositions of the present invention to the cell sample or tissue sample; and 3) determining the amount of the regulation of the one or more energy biomarkers after administering the one or more compounds or compositions compared to the state of the energy biomarkers before administering the one or more compounds or compositions. Another such test or assay would involve 1) obtaining a cell sample or tissue sample from a subject in which modulation of one or more energy biomarkers can be determined; 2) administering to the cell sample or tissue sample at least two compounds or compositions of the invention; 3) determining the amount of modulation of the one or more energy biomarkers after administration of the at least two compounds or compositions as compared to the state of the energy biomarkers prior to administration of the at least two compounds or compositions; and 4) selecting a compound or composition for treatment, inhibition, or modulation based on the amount of modulation determined in step 3.

[0203] In some embodiments, the present invention provides methods of using Type I-VI polymorphs to treat or prevent damage or injury caused by radiation exposure, and methods of using such compounds to treat or prevent damage or injury caused by radiation exposure. In some embodiments, a method for treating or preventing damage or injury caused by radiation exposure, the method comprising administering a therapeutically effective amount or a prophylactically effective amount of the Type I-VI polymorph or composition of the present invention to a cell or cell type, a tissue or tissue type, or a subject in need. In one embodiment, the Type I-VI polymorph is used for treatment during, or after, or during and after radiation exposure. In another embodiment, the Type I-VI polymorph is used for treatment before radiation exposure. In another embodiment, the Type I-VI polymorph is administered simultaneously during radiation exposure. In another embodiment, the one or more compounds are administered after radiation exposure.

[0204] In some embodiments, the present invention provides methods of using Type I-VI polymorphs to treat damage or injury caused by radiation exposure, and methods of using such compounds to treat or prevent damage or injury caused by radiation exposure. In some embodiments, a method for treating damage or injury caused by radiation exposure, the method comprising administering a therapeutically effective amount or a prophylactically effective amount of the Type I-VI polymorph or composition of the present invention to a cell or cell type, a tissue or tissue type, or a subject in need. In one embodiment, the Type I-VI polymorph is used for treatment during, or after, or during and after radiation exposure. In another embodiment, the Type I-VI polymorph is used for prevention before radiation exposure. In another embodiment, the Type I-VI polymorph is administered simultaneously during radiation exposure. In another embodiment, the one or more compounds are administered after radiation exposure.

[0205] Pharmaceutical preparations

[0206] The compound or composition of the present invention can be formulated into a pharmaceutical composition by preparing additives, such as pharmaceutically acceptable adjuvants, pharmaceutically acceptable carriers, and pharmaceutically acceptable vehicles. Suitable pharmaceutically acceptable adjuvants, carriers, and vehicles include processing aids and drug delivery regulators and reinforcing agents, for example, calcium phosphate, magnesium stearate, talcum powder, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, glucose, hydroxypropyl-β-cyclodextrin, polyvinyl pyrrolidone, low melting point waxes, ion exchange resins, etc., and any two or more combinations thereof. Other suitable pharmaceutically acceptable excipients are described in “Remington's Pharmaceutical Sciences,” Mack Pub. Co., New Jersey (1991), and “Remington: The Science and Practice of Pharmacy,” Lippincott Williams & Wilkins, Philadelphia, 20th edition (2003) and 21st edition (2005), which are incorporated herein by reference.

[0207] The pharmaceutical composition may comprise a unit dose formulation, wherein the unit dose is a dose sufficient to have a therapeutic or inhibitory effect, or an amount effective to modulate, normalize, or enhance an energy biomarker. The unit dose may be an amount sufficient to have a therapeutic or inhibitory effect, or an amount effective to modulate, normalize, or enhance an energy biomarker as a single dose. Alternatively, the unit dose may be a dose that is administered periodically during the course of treating or inhibiting a condition, or a dose that is administered periodically for modulating, normalizing, or enhancing an energy biomarker.

[0208] The pharmaceutical composition containing the compounds or compositions of the present invention can be in any form suitable for the intended method of administration, including, for example, solutions, suspensions or emulsions. Liquid carriers are generally used to prepare solutions, suspensions or emulsions. Liquid carriers that can be considered for the practice of the present invention include, for example, water, saline, pharmaceutically acceptable organic solvents, pharmaceutically acceptable oils or fats, etc., and mixtures of two or more thereof. The liquid carrier may include other suitable pharmaceutically acceptable additives, such as solubilizers, emulsifiers, nutrients, buffers, preservatives, suspending agents, thickeners, viscosity regulators, stabilizers, etc. Suitable organic solvents include, for example, monohydric alcohols, such as ethanol, and polyols, such as glycols. Suitable oils include, for example, soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, etc. For parenteral administration, the carrier may also be an oil ester, such as ethyl oleate, isopropyl myristate, etc. The composition of the present invention may also be in the form of microparticles, microcapsules, liposome encapsulations, etc., and any combination of two or more thereof.

[0209] Time-release or controlled-release delivery systems can be used, such as diffusion-controlled matrix systems or erodible systems, for example, as described in Lee, "Diffusion-Controlled Matrix Systems", pp. 155-198 and Ron and Langer, "Erodible Systems", pp. 199-224, and "Treatise on Controlled Drug Delivery", A. Kydonieus Ed., Marcel Dekker, Inc., New York 1992. The matrix can be, for example, a biodegradable material that can degrade spontaneously in situ and in vivo, for example, by hydrolysis or enzymatic cleavage, for example, by proteases. The delivery system can be, for example, a naturally occurring or synthetic polymer or copolymer, for example in the form of a hydrogel. Exemplary polymers with cleavable linkages include polyesters, polyorthoesters, polyanhydrides, polysaccharides, poly(phosphates), polyamides, polyurethanes, poly(imidocarbonates) and poly(phosphazenes).

[0210] The compounds of the present invention or compositions can be used as needed by enteral, oral, parenteral, sublingual, inhalation (for example, as mist or spray), rectum, or locally containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants and vehicle dosage unit formulations. For example, suitable modes of administration include oral, subcutaneous, transdermal, mucosal, ion introduction, vein, artery, intramuscular, intraperitoneal, intranasal (such as through nasal mucosa), subdural, rectum, gastrointestinal tract, etc., and directly to a specific or affected organ or tissue for administration. In order to be delivered to the central nervous system, spinal cord and epidural administration or administration to the ventricle can be used. Topical administration can also involve the use of transdermal administration such as transdermal patches or ion permeation devices. The term parenteral used in the present invention includes subcutaneous injection, intravenous, intramuscular, intrasternal injection or infusion technology. The compound or composition is mixed with a pharmaceutically acceptable, suitable carrier, adjuvant and vehicle for the desired route of administration. Oral administration is a preferred route of administration, and preparations suitable for oral administration are preferred preparations. The compounds used in the present invention can be administered in solid form, liquid form, aerosol form or in tablets, pills, powder mixtures, capsules, granules, injections, creams, solutions, suppositories, enemas, colon lavages, emulsions, dispersions, food premixes and other suitable forms. The compound or composition can also be administered in liposome formulations. The compound can also be administered as a prodrug, wherein the prodrug undergoes conversion into a therapeutically effective form in the treated subject. Other methods of administration are known in the art.

[0211] In some embodiments of the invention, particularly those embodiments, wherein the formulation is for injection or other parenteral administration, including the routes listed herein, but also including routes for oral, gastric, gastrointestinal, or enteral administration, the formulations and formulations used in the methods of the invention are sterile. Sterile pharmaceutical preparations are compounded or prepared according to pharmaceutical grade sterilization standards known to those skilled in the art (U.S. Pharmacopeia Chapters 797, 1072, and 1211; California Business and Professions Code 4127.7; 16 California Statutes 1751, 21 Federal Regulations 211).

[0212] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenteral acceptable diluent or solvent, such as, for example, a propylene glycol solution. The acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspension media. For this reason, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can be used to prepare injections.

[0213] Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose, or starch. Such dosage forms may also contain other substances in addition to the inert diluent, for example, lubricants such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer. Tablets and pills may also be prepared with enteric coatings.

[0214] Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also include adjuvants, such as wetting agents, emulsifying and suspending agents, cyclodextrins, and sweeteners, flavoring agents, and perfuming agents.

[0215] The compounds or compositions of the present invention can also be administered in the form of liposomes. As is well known in the prior art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by monolayer or multilayer hydrated liquid crystals dispersed in an aqueous medium. Any non-toxic, physiologically acceptable and metabolizable lipids capable of forming liposomes can be used. The compositions of the present invention in the form of liposomes may contain stabilizers, preservatives, excipients, etc. in addition to the compounds of the present invention. Preferred lipids are natural and synthetic phospholipids and phosphatidylcholines (lecithins). Methods for forming liposomes are well known in the art, see, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, NW, p. 33 et seq (1976).

[0216] The present invention also provides products and kits containing materials for treating or inhibiting oxidative stress disorders. The present invention also provides kits containing any one or more compounds or compositions described herein. In some embodiments, the kits described herein contain a suitable container.

[0217] In other aspects, the kits can be used in any of the methods described herein, including, for example, treating an individual with a mitochondrial disease, or inhibiting a mitochondrial disease in an individual.

[0218] The amount of active ingredient that can be combined with carrier materials to produce a single dosage form will vary according to the host and specific mode of administration to which the active ingredient is applied. However, it should be understood that the specific dosage level of any particular patient will depend on a variety of factors, including the activity of the specific compound used, patient age, body weight, body area, body mass index (BMI), general health, sex, diet, administration time, route of administration, excretion rate, drug combination, and the type, progression, and severity of the specific disease being treated. The selected drug unit dose is usually formulated and administered to provide a determined drug final concentration in the blood, tissue, organ, or other target area of ​​the body. The therapeutically effective amount or effective amount in a given scenario can be easily determined by routine experiments, and is within the skills and judgment of an ordinary clinician.

[0219] Examples of dosages that can be used are in a dosage range of about 0.1 mg / kg to about 300 mg / kg body weight, or about 1.0 mg / kg to about 100 mg / kg body weight, or about 1.0 mg / kg to about 50 mg / kg body weight, or about 1.0 mg / kg to about 30 mg / kg body weight, or about 1.0 mg / kg to about 10 mg / kg body weight, or about 10 mg / kg to about 100 mg / kg body weight, or about 50 mg / kg to about 150 mg / kg body weight, or about 100 mg / kg to about 200 mg / kg body weight, or about 150 mg / kg to about 250 mg / kg body weight, or about 200 mg / kg to about 300 mg / kg body weight, or about 250 mg / kg to about 300 mg / kg body weight. The compounds or compositions of the present invention can be administered in a single daily dose, or the total daily dose can be administered in divided doses twice, three times or four times a day.

[0220] Although the compounds or compositions of the present invention can be administered as the sole active drug, they can also be used in combination with one or more other drugs for treating or inhibiting the condition. Representative drugs used in combination with the compounds or compositions of the present invention for treating or inhibiting mitochondrial diseases include, but are not limited to, coenzyme Q, vitamin E, idebenone, MitoQ, vitamins, NAC, and antioxidant compounds.

[0221] When additional active drugs are used in combination with the compounds or compositions of the present invention, the additional active drugs can generally be used in therapeutic amounts as described in the Physicians' Desk Reference (PDR), 53rd Edition (1999), or in therapeutically useful amounts known to those of ordinary skill in the art.

[0222] The compounds or compositions of the present invention and other therapeutically active drugs can be administered at the recommended maximum clinical dose or at a lower dose. The dosage level of the active compound in the composition of the present invention can be changed so as to obtain the desired therapeutic response according to the route of administration, the severity of the disease and the patient's response. When administered in combination with other therapeutic agents, the therapeutic agents can be formulated into separate compositions that are administered at the same time or at different times, or the therapeutic agents can be administered in a single composition.

[0223] The present invention will be further understood by way of the following non-limiting examples.

[0224] Preparation of the composition of the present invention

[0225] The compositions of the present invention can be prepared from readily available starting materials using the following general methods and procedures. It should be understood that where typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.) are given, other process conditions may also be used unless otherwise stated. Optimum reaction conditions may vary with the specific reactants or solvents used, but those skilled in the art can determine these conditions by conventional optimization procedures.

[0226] Synthesis reaction parameters

[0227] Solvents used to synthesize the compounds and compositions of the present invention include, for example, methanol ("MeOH"), acetone, water, acetonitrile, 1,4-dioxane, dimethylformamide ("DMF"), benzene, toluene, xylene, tetrahydrofuran ("THF"), chloroform, methylene chloride (dichloromethane, ("DCM")), ether, pyridine, 2-methyltetrahydrofuran ("2-MeTHF"), dimethylacetamide ("DMA"), ethyl acetate ("EtOAc"), ethanol ("EtOH"), isopropanol ("IPA"), isopropyl acetate ("IPAc"), methylcellulose ("MC"), acetonitrile ("MeCN"), methanol (MeOH), methyl tert-butyl ether ("MTBE"), phosphate buffered saline ("PBS"), tetrahydrofuran ("THF"), etc., and mixed solvents thereof.

[0228] The term "qs" refers to the addition of an amount sufficient to achieve a stated function, eg, to bring a solution to a desired volume (ie, 100%).

[0229] The compounds of this invention and compositions are obtained by the combination synthesis of suitable generally known synthetic methods. The technology used to synthesize the compounds of this invention and compositions is according to the teachings of the present invention, is obvious to the technicians of the relevant field and can be easily obtained. The following discussion is provided to illustrate some different methods that can be used to prepare the compounds of this invention and compositions. However, the discussion is not intended to limit the scope of the reaction or reaction sequence that can be used to prepare the compounds of this invention and compositions.

[0230] Other methods for producing the compounds and compositions of the present invention will be apparent to those skilled in the art based on the teachings of the present invention. Example

[0231] Example 1. Synthesis of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (Type I)

[0232] Example 1A. Extraction of (1S,2S)-(+)-pseudoephedrine free base

[0233] To a suspension of (1S,2S)-(+)-pseudoephedrine hydrochloride (300 g, spectrally pure) in 2-MeTHF (1.5 L, 5 vols) was added 20% aqueous NaOH (750 mL, 2.5 vols) and the resulting mixture was stirred for 30 min (some solids remained undissolved) and then transferred to a separatory funnel. The resulting lower aqueous layer was drained along with the solids remaining in the interphase and then extracted with 2-MeTHF (750 mL, 2.5 vols) to completely dissolve the undissolved solids to form two clear layers. The combined organic layers were evaporated to dryness on a rotary evaporator and the resulting solids were dried in a vacuum oven at 50°C overnight to give 240.3 g of the free base as a white solid (97.7% recovery).

[0234] Example 1B. Precipitation of (1S,2S)-pseudoephedrine from 2-MeTHF / heptane

[0235] (1S,2S)-pseudoephedrine (Sigma-Aldrich, sku#212464, 8.2 g) was dissolved in 2-MeTHF (41 ml, 5 volumes) at 50°C. The resulting solution was diluted with heptane (82 ml, 10 volumes) and the resulting suspension was stirred and reacted overnight at room temperature. The crystallized (1S,2S)-pseudoephedrine was filtered and then dried under vacuum at 40°C overnight to give 6.4 g (78%) of white crystalline material. The filtrate was discarded into general waste.

[0236] The relatively low (78%) crystallization yield prompted additional crystallization experiments using a higher ratio of heptane to 2-MeTHF. The crystalline (1S,2S)-pseudoephedrine obtained in the above experiment was dissolved in 2-MeTHF (32 ml, 5 volumes) at 50°C. The resulting solution was diluted with heptane (32 ml, 5 volumes) and the resulting suspension was chased with heptane (3×50 ml) on a rotary evaporator until the molar ratio of 2-MeTHF to heptane was less than 6% by NMR analysis. The resulting suspension was filtered and the product was dried in vacuo at 40°C overnight to give 6.3 g (98%) of white crystalline material.

[0237] Example 1C. Chiral Resolution of Water-Soluble Vitamin E (Trolox) Using (1S,2S)-(+)-Pseudoephedrine

[0238]

[0239] Water-soluble vitamin E (Trolox, 316.6 g, 1.27 mol) and (1S,2S)-(+)-pseudoephedrine free base (240.0 g, 1.46 mol) described in Example 1A were charged to a 4 L jacketed reactor equipped with an overhead stirrer, temperature probe, and nitrogen vent. Ethyl acetate (EtOAc, 1585 mL, 5 volumes) was then charged and the resulting slurry was heated to 50° C. to give a clear solution. (At 40° C., premature precipitation of (R)-water-soluble vitamin E-pseudoephedrine salt (before complete dissolution of the water-soluble vitamin E racemate) was sometimes observed. If premature precipitation occurred, the reaction mixture was heated (usually to reflux temperature) to achieve complete dissolution). The reaction mixture was cooled overnight to room temperature, at which time a large amount of precipitation was observed. The mixture was cooled to 10° C. over 30 minutes and maintained at this temperature for 1 hour. The solid formed was collected by filtration, the wet cake was washed with EtOAc (1.9 L, 6 volumes), and then the cake was dried in a vacuum oven at 25-30° C. to constant weight to give 188.1 g (71.3%, based on (R)-water-soluble vitamin E (Trolox)) of a white solid. Chiral HPLC data indicated nearly 100% enantiomeric purity.

[0240] Example 1D. Recovery of (R)-Water-soluble Vitamin E from Its Salt Using (1S,2S)-(+)-Pseudoephedrine

[0241]

[0242] R-Water-soluble Vitamin E PE Salt

[0243] The obtained (R)-water-soluble vitamin E PE salt (187.3g, 0.45mol) is charged into a 2L round-bottom flask, followed by the addition of 2-MeTHF (570ml, 3 volumes) to form a slurry. Hydrochloric acid (2.5M, 325ml, 0.81mol, 1.75 equivalents) is added in batches, while keeping the temperature below 25°C. The water-soluble vitamin E-PE salt is dissolved, and the (R)-water-soluble vitamin E is extracted into the organic phase. Small black fragments are observed at the interface and are kept together with the water layer. The obtained aqueous phase is further extracted with 2-MeTHF (2×200ml). Then, the combined organic layer is washed with 15% NaCl (200ml) and then with water (200ml). The organic layer is dried with anhydrous sodium sulfate (150g), filtered, and evaporated to obtain a white solid, which is dried at constant weight in a vacuum oven at 30°C to obtain 128.3g, which is a superstoichiometric amount.

[0244] Example 1E. Preparation of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide

[0245]

[0246] CDI (Sigma-Aldrich) (188 g, 1.16 mol) was charged into a 3-neck 2L RBF (round bottom flask) equipped with an overhead stirrer, nitrogen inlet, and temperature probe. 2-MeTHF (290 mL) was added to form a stirrable slurry, followed by the slow addition of a solution of (R)-water-soluble vitamin E (126.0 g, 504 mmol) in 2-MeTHF (500 ml) at 30°C. A slightly exothermic reaction with CO2 evolution was observed. Degassing began after the addition of approximately one-third of the (R)-water-soluble vitamin E. Complete dissolution of the starting material was observed within approximately 15 minutes.

[0247] The contents of the flask were slowly added to 28-30% aqueous ammonia (380 ml) precooled to 5° C., keeping the temperature below 30° C. The resulting biphasic suspension was stirred at room temperature and monitored by HPLC. The reaction was complete after 36 h and was then further processed after 48 h.

[0248] The reaction mixture was acidified to pH 1-2 with sulfuric acid (1:4 v / v) (850 ml), and the temperature was maintained at ≤ 28 ° C. The reaction released a large amount of heat. The aqueous layer (pH = 1) was removed, and the resulting organic layer was washed with NaCl (15% aqueous solution w / v, 250 mL), NaHCO3 (1 M, 250 mL), NaCl (15% aqueous solution w / v, 250 mL) and water (250 ml). Most of the organic layer was used in the subsequent steps.

[0249] Example 1F. Preparation of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0250]

[0251] A 2L 3N RBF (round bottom flask) equipped with an overhead stirrer and a thermocouple was charged with a solution of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide (708 ml) containing -0.39 mole of the amide intermediate and water (126 ml).

[0252] A stock solution of FeCl3×6H2O (480 g, 1.78 mol) in water (336 ml) was divided into 4 equal portions (204 g each) and one quarter of the iron (III) chloride solution was added to the reaction flask. A weak (~3°C) exotherm was observed and the color of the organic layer turned nearly black and then lightened to dark brown. The biphasic reaction mixture was stirred vigorously at room temperature for 40 minutes. After removing the slightly discolored aqueous phase, another portion of the iron (III) chloride solution was added and the reaction was stirred for 40 minutes. The operation was repeated one or more times and the resulting organic phase was stored overnight at room temperature. The fourth treatment with FeCl3×6H2O was performed the next morning. Nearly complete (99.44%) conversion of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide to (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide was observed. Initial iron extraction was accomplished with 1M trisodium citrate solution (2 x 350 ml); the AUC% of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide was increased to 0.84%. The pH of the organic phase was maintained at a strongly acidic pH = 1. A 1 ml aliquot of the organic phase was treated with 1M NaHCO3, which resulted in a large red Fe(OH)3 precipitate. Based on this observation, another trisodium citrate (175 ml) wash was performed (0.74% (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide). The experiment was repeated with a 1 ml aliquot with 1M NaHCO3, and no precipitation was obtained in the aqueous layer, which was yellow in color instead of red, indicating that the iron had been completely or nearly completely removed.

[0253] The organic layer was heated to 40°C to prevent premature precipitation of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and then washed with 1 M sodium bicarbonate solution (175 ml). Phase separation was not immediate but the formation of two clear yellow layers was complete within 15 minutes. The organic layer (0.30% of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide) was further washed with water (350 ml) to give 0.22% of (R)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxamide. The organic layer was evaporated to give 96 g of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0254] The combined bicarbonate / water layers were re-extracted with 2 x 250 ml of 2-MeTHF. Evaporation of these extracts gave 4.0 and 0.9 g of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, respectively.

[0255] The combined solids (100.9 g crude (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, 84% yield based on (R)-water-soluble vitamin E-pseudoephedrine salt) were dissolved in isopropanol (600 ml) at 70°C and the resulting yellow solution was charged to a 2L 3NRBF (three-necked round bottom flask) equipped with an overhead stirrer, heating mantle and thermocouple.

[0256] Heptane (600 ml) was added and no precipitation was observed. The reaction mixture was reheated to 55°C and slowly cooled to room temperature. Seed crystals of the desired polymorph (0.2 g) were added and the reaction mixture was stirred at room temperature overnight. A large amount of precipitation was observed overnight. The reaction mixture was then cooled to 7°C and stirring was continued for 8 hours. The resulting product was filtered, washed with isopropanol-heptane 1:1 v / v (2×75 ml) and dried at 40°C for one week. The yield was 69.4 g of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (58% based on (R)-water-soluble vitamin E-pseudoephedrine salt). The XRPD data of the product corresponded to the desired Type I polymorph.

[0257] Example 2. Solubility determination of mode A

[0258] (R)-water-soluble vitamin E is prepared by twice splitting racemic water-soluble vitamin E with methylbenzylamine in a manner similar to that described in Example 29 of U.S. Patent No. 4,026,907. The (R)-water-soluble vitamin E is used to synthesize the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide starting material. The starting material is referred to as Mode A and is used for the solubility determination.

[0259] Excess solids were slurried in 17 solvent systems with different properties for at least 3 days. The slurry was centrifuged and the resulting clear solution was used for gravimetric analysis. The compound showed improved solubility at ambient temperature in MeOH, EtOH, and at 50°C in IPA, acetone, MeOH, EtOH and 2-MeTHF. Moderate solubility was observed in PBS of EtOAc, THF, IPA, acetone, 2-MeTHF, MeCN, 0.5% methylcellulose / 2% Tween 80, IPAc and 4% DMA at ambient conditions and 50°C. Limited solubility was observed in aqueous solutions of heptane, toluene, MTBE, water and 0.5% methylcellulose at ambient conditions and elevated temperatures. Table 9 shows the solubility data for the assay. A ± 10% error is expected.

[0260] Table 9: Solubility of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in various solvent systems (starting material, use mode A)

[0261]

[0262] *Some concentrations involve solvent components

[0263] As shown in Table 9, Pattern A is soluble in various polar and non-polar solvents and has increased solubility in a simple detergent (0.5% MC / 2% Tween 80).

[0264] Example 3. Short-term pulping experiment

[0265] Short-term pulping experiments of the starting material pattern A were carried out for at least 3 days in 17 different solvent systems with different properties at two different temperatures (25°C and 50°C). A 48-position parallel synthesizer (Chemglass) reaction block was used to heat and stir the slurry in a 2 mL HPLC vial. After an appropriate time, the vial was centrifuged and the wet solid was used for X-ray diffraction. Table 10 shows the results of the pulping experiments. These results show that the pattern A solid is relatively stable if pulped in most of these solvents for a short period of time. However, pulping pattern A in THF and 0.5% methylcellulose / 2% Tween 80 gave two new X-ray patterns, referred to as patterns B and C ( Figure 1 ).

[0266] Table 10: Summary of at least 3 days of slurrying of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (starting material, using mode A)

[0267]

[0268] Example 4. Evaporation crystallization experiment

[0269] Evaporative crystallization experiments of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide were performed using samples generated by gravimetric solubility determination (Example 2). XRPD analysis of most samples resulted in pattern A. However, it was found that XRPD analysis of the solid isolated from 2-MeTHF at 25°C provided a unique crystallization pattern, designated pattern D, as shown in Figure 2XRPD analysis of samples obtained from IPAc, 0.5% MC in water and 4% DMA in PBS yielded mainly amorphous patterns, except for a crystalline form consistent with pattern A obtained from IPAc at 50°C. All results are summarized in Table 11.

[0270] Table 11: Summary of the evaporation crystallization experiment of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (starting material, using mode A)

[0271]

[0272]

[0273] NA - Drying was not possible due to sample drying overnight

[0274] Example 5. Crystallization experiment

[0275] Fast and slow cooling single solvent crystallization experiments were conducted in toluene, EtOAc, IPA, acetone, EtOH, 2-MeTHF, and IPA containing 2% water (Table 12). A 48-position parallel synthesizer (Chemglass) reaction block was used for heating and stirring in a 4mL vial. Each vial was loaded with 50-80mg of the starting material (mode A) and equipped with a magnetic stirring bar. The main solvent was added and stirred and heated until dissolved. Once completely dissolved, the sample was slowly cooled by radiation cooling or rapidly cooled using an ice bath, followed by stirring and balancing overnight. For the binary solvent crystallization method, an anti-solvent (heptane) was added in two ways (Table 13). Method one, the anti-solvent was added dropwise to the sample solution until a slight precipitation was observed. Method two, the sample solution was added in reverse to the heated anti-solvent, wherein the ratio of the sample solution to the heated anti-solvent was 2: 1, and then cooled. The solid sample obtained after overnight equilibration was separated by filtration, and the unprecipitated sample was evaporated under a gentle nitrogen stream. All samples were dried in a vacuum oven overnight under ambient conditions and analyzed using XRPD to check for product form changes. All experimental details and results are summarized in Tables 12-13.

[0276] XRPD analysis of all isolated solids indicated that the resulting crystalline form was nearly identical to the starting material Pattern A. However, single solvent crystallization methods in 2-MeTHF using both rapid and slow cooling methods did not observe solids upon cooling and evaporation to dryness under nitrogen. The XRPD patterns of the solids obtained by these evaporations were consistent with the previously observed Pattern D ( Figure 3 ). Rapid cooling crystallization in EtOAc produced a unique crystalline solid as described by XRPD, which was compared with all known forms and was designated Pattern E ( Figure 4 ). Binary solvent crystallizations in EtOH / heptane and acetone / heptane using fast and slow cooling methods yielded XRPD patterns consistent with Pattern A. Table 12: Summary of single solvent crystallization experiments of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (starting material, using Pattern A)

[0277]

[0278] Table 13: Summary of Binary Solvent Crystallization Experiments of (R)-2-Hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide (Starting Material, Using Mode A)

[0279]

[0280]

[0281] Example 6. Scale-up experiment

[0282] Scale-up experiments were performed in 300 mg batches by single solvent rapid cooling crystallization in 2-MeTHF, EtOAc, and slurrying in THF and 0.5% methylcellulose / 2% Tween 80 in an attempt to separate the previously observed modes D, E, B, and C for further characterization. The experimental details and results are summarized in Tables 14-15.

[0283] Rapid cooling single solvent crystallization experiments were conducted in 2-MeTHF and EtOAc (Table 14). A 48-position parallel synthesizer (Chemglass) reaction block was used to heat and stir in a 4 mL vial. Each vial was loaded with 300 mg of the starting material and equipped with a magnetic stirring bar. 6 mL of the main solvent was added and heated to 60 ° C with stirring until dissolved. Once completely dissolved, the sample was transferred to an ice bath (rapid cooling) and inoculated with a spatula tip of mode D or E, followed by stirring and balancing at room temperature overnight. The solid sample obtained after overnight equilibration was separated by filtration, and the unprecipitated sample was evaporated under a gentle nitrogen stream. All samples were dried overnight under vacuum under ambient conditions, and XRPD analysis was performed to check product form changes.

[0284] Pulping experiments were performed in THF and 0.5% methylcellulose / 2% Tween 80 (Table 15). Heating and stirring were performed in 4 mL vials using a 48-position parallel synthesizer (Chemglass) reaction block. Each vial was charged with 300 mg of starting material and equipped with a magnetic stir bar. Up to 2 mL of slurry solvent was added under ambient conditions and allowed to equilibrate for 30 minutes before adding a spatula tip of Mode B or C.

[0285] XRPD analysis of the solid isolated by single solvent crystallization using a rapid cooling method in 2-MeTHF showed a unique XRPD pattern, referred to as pattern F ( Figure 4 ). XRPD analysis of the crystalline solid obtained by rapid cooling crystallization in EtOAc showed that it was consistent with pattern E ( Figure 5 ). Pulping experiments in THF and 0.5% methylcellulose / 2% Tween 80 yielded patterns B and C after 24 hours of equilibration, respectively. Figure 6-7 ).

[0286] Table 14: Summary of single solvent crystallization scale-up experiments for (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0287]

[0288]

[0289] Table 15: Summary of pulping scale-up experiments for (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0290]

[0291] Example 7. Competitive pulping

[0292] Competitive pulping of patterns A, B, C, D, E and F was performed under ambient conditions in aqueous solutions of IPA, IPA / 2% water and 0.5% methylcellulose, as shown in Table 16. Approximately 100 mg of the starting material, pattern A, was added to a glass vial equipped with a magnetic stir bar. Solvent was added to the vial and allowed to slurry with pattern A for 15 minutes, then approximately 10-20 mg of each opposing pattern (B, C, D, E and F) was added to each vial. The samples were allowed to equilibrate under stirring and after 24 hours or equilibration, XRPD analysis showed that the solids isolated from the pulping in the 0.5% methylcellulose aqueous solution were a mixture of patterns A / C ( Figure 8 All other solids isolated from IPA and IPA / 2% water were consistent with starting material pattern A ( Fig. 9 However, after 7 days of equilibration, complete conversion to Mode C was observed in pulping from 0.5% methylcellulose in water ( Figure 8 ), where the solids separated by competitive pulping from IPA and IPA / 2% water were consistent with Mode A ( Fig. 9 Table 16: Summary of competitive pulping experiments of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in 2 mL solvent

[0293]

[0294] *- Used to saturate the solvent and make a slurry of Mode A

[0295] Example 8. High Humidity Study

[0296] All patterns A, B, C, D, E and F were subjected to a 7-day high moisture humidity experiment (Table 17) under ambient conditions, >95% RH (relative humidity). Approximately 30 mg of each pattern was weighed into a 4 ml glass vial. The uncapped 4 ml vial was then inserted into a 20 ml scintillation vial filled with water and capped. After 24 hours of equilibrium, visual inspection was performed to check the changes in physical appearance, but no changes were observed. After 7 days of equilibrium at high humidity, the sample showed no physical changes and was analyzed by XRPD to check the sample form. After 7 days of equilibrium, patterns A, C and E showed no changes in form; however, pattern B was converted to a mixture of pattern A / B. At >95% RH, it was found that pattern D was converted to a mixture of pattern D / B, and pattern F was converted to pattern E. All experimental details and results are summarized in Table 17.

[0297] Table 17: Summary of 7-day high moisture experiment of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0298]

[0299] Example 9. Grinding experiment

[0300] Pyruvate dehydrogenase deficiency; The tests were performed using a mortar and pestle in IPA and water dry grinding and solvent drop grinding (Table 18). After mild grinding mode A, no change in the crystal form was observed by XRPD analysis.

[0301] Table 18: Summary of grinding experiments for (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0302]

[0303]

[0304] Example 10. Water Solubility

[0305] The water solubility of patterns A, B, C, E and F is detected using an Agilent HPLC system. Approximately 10-20 mg of each form is loaded into a 2 mL glass vial equipped with a magnetic stirring bar and 2 mL of water is added. The sample is stirred overnight at ambient conditions. After 24 hours of equilibrium, the sample is centrifuged and poured into an HPLC sample bottle. A calibration curve is generated based on pattern A in 0.05, 0.1, 0.5 and 1.0 mg / mL of MeOH. After the injection standard curve, the sample is run as is. All experimental details and results are summarized in Table 19.

[0306] Table 19: Summary of water solubility experiments of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide

[0307]

[0308] Example 11. Characterization of Forms

[0309] The solid form of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide was characterized by XRPD, DSC, 1 H NMR, TGA, Karl Fischer, optical microscopy and moisture sorption were performed. The results are summarized in Table 1.

[0310] Mode A (anhydrous, type I)

[0311] Using the starting material as described in Example 2, XRPD analysis of the yellow starting material was found to give a crystalline form designated Pattern A ( Fig.10 The birefringence observed by optical microscopy confirmed the crystallinity observed by XRPD. Fig.11 As shown, the morphology of the crystals was determined to be irregular shapes with certain aggregation.

[0312] Thermal analysis by DSC showed a single endothermic peak at 152.9 °C, which then degenerated after 200 °C ( Fig.12 ).

[0313] TGA analysis showed no weight loss between 45 and 160 °C, however, weight loss due to decomposition was observed from 160 to 300 °C ( Fig.13 The minimum water content was confirmed by Karl Fischer analysis, which showed that the material contained approximately 0.12 wt% water.

[0314] 1Further analysis by HNMR showed that the starting material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and contained 0.28 wt% residual IPA. Fig.40 .

[0315] The hygroscopic analysis of the starting material was done by equilibrating the sample at 25°C and 50% RH to simulate ambient laboratory conditions. The humidity was then dropped to 0% RH, increased from 0 to 95% RH, dropped from 95 to 0% RH, increased from 0 to 95% RH, and then dropped from 95 to 50% RH. Each point represents the estimated asymptotic weight at each humidity or weight. The starting material was found to be non-hygroscopic, absorbing 0.1% of water at 90% RH. No hysteresis was observed upon desorption ( Fig.14 ). The XRPD analysis of the sample after moisture sorption analysis was found to be consistent with Pattern A of the starting material.

[0316] Mode E (anhydrate, type II)

[0317] Pattern E (anhydrate, Form II) was observed during single solvent crystallization using a 50 mg batch, rapid cooling method, and again in a 300 mg scale-up (Examples 5 and 6). From XRPD analysis of the solid, it was found that a single crystalline form was obtained, designated Pattern E ( Fig.15 The birefringence observed by optical microscopy confirmed the crystallinity observed by XRPD. Fig.16 As shown, the morphology of the crystals was determined to be irregular shapes with certain aggregation.

[0318] Thermal analysis of a 50 mg batch by DSC showed two endothermic peaks at 133.9 °C and 151.3 °C, followed by degradation after 200 °C ( Fig.17 ).

[0319] TGA analysis of a 50 mg batch showed a 0.4% weight loss between 120-140 °C, probably due to loss of EtOAc followed by decomposition ( Fig.18 The minimum water content was confirmed by Karl Fischer analysis, which showed that the material contained approximately 0.1 wt. % water.

[0320] Model E (from the 300 mg scale-up) 1 Further analysis by HNMR showed that the material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and contained 0.4 wt% residual EtOAc. Fig.44 .

[0321] The hygroscopic analysis of Pattern E (from the 300 mg scale-up) was done by equilibrating the sample at 25°C and 50% RH to simulate ambient laboratory conditions. The humidity was then dropped to 0% RH, increased from 0 to 95% RH, dropped from 95 to 0% RH, increased from 0 to 95% RH, and then dropped from 95 to 50% RH. Each point represents the estimated asymptotic weight at each humidity or weight. Pattern E was found to be non-hygroscopic, absorbing 0.2% water at 95% RH. No hysteresis was observed upon desorption ( Fig.19 ). It was found that the XRPD analysis of the sample after moisture absorption analysis was consistent with the described pattern E.

[0322] Mode C (hydrate, type III)

[0323] Pattern C (hydrate, Form III) was observed in a 50 mg batch, during short-term slurrying in 0.5% methylcellulose / 2% Tween 80, and again in a 300 mg scale-up batch (Examples 3 and 6, respectively). From XRPD analysis of the solid, it was found that a single crystalline form was obtained, designated Pattern C ( Fig. 20 The birefringence observed by optical microscopy confirmed the crystallinity observed by XRPD. Fig.21 As shown, the morphology of the crystals was determined to be irregular shapes with certain aggregation.

[0324] Thermal analysis of a 50 mg batch by DSC showed two endothermic peaks at 72 °C and 150.7 °C, followed by degradation after 200 °C ( Fig. 22 ).

[0325] TGA analysis of a 50 mg batch showed a 2.5% weight loss between 20-60°C, followed by a 2.3% weight loss between 60-125°C, probably due to dehydration followed by decomposition ( Fig.23 ). The water content was confirmed by Karl Fischer analysis, which showed that the material contained about 4.3 wt% water, slightly less than the monohydrate.

[0326] Model C (300mg scale-up) 1 Further analysis by H NMR showed that the material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. Fig.42 .

[0327] The hygroscopic analysis of mode C (300 mg scale-up) was done by equilibrating the sample at 25°C and 50% RH to simulate ambient laboratory conditions. The humidity was then dropped to 0% RH, increased from 0 to 95% RH, dropped from 95 to 0% RH, increased from 0 to 95% RH, and then dropped from 95 to 50% RH. Each point represents the estimated asymptotic weight at each humidity or weight. Mode C was found to be slightly hygroscopic, absorbing 2% water at 95% RH. This increased the total water content to about 6%, which is the water content of a monohydrate. However, upon reducing the relative humidity, the solid lost water. Therefore, this may be a channel hydrate. No hysteresis was observed upon desorption ( Fig.24 ). It was found that the XRPD analysis of the sample after moisture absorption analysis was consistent with the pattern C.

[0328] Pattern B (THF solvate, Type IV)

[0329] Pattern B (THF solvate, Form IV) was observed in a 50 mg batch, during short-term slurrying in THF, and again in a 300 mg scale-up (Examples 3 and 6, respectively). From XRPD analysis of the solid, it was found that a single crystalline form was obtained, referred to as Pattern B ( Fig.25 The birefringence observed by optical microscopy confirmed the crystallinity observed by XRPD. Fig.26 As shown, the morphology of the crystals was determined to be needle-shaped with certain aggregation.

[0330] Thermal analysis of a 50 mg batch by DSC showed three endothermic peaks at 70.5 °C, 89.1 °C and 149.7 °C, followed by degradation after 200 °C ( Fig. 27 ).

[0331] TGA analysis of a 300 mg batch showed a 4.7% weight loss between 25 and 115 °C, probably due to the loss of THF followed by decomposition ( Fig.28 ). The water content was confirmed by Karl Fischer analysis, which showed that the material contained about 0.3 wt% water.

[0332] Model B (300mg scale-up) 1 Further analysis by H NMR showed that the material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and contained 6.9 wt% residual THF. Fig.41 .

[0333] The hygroscopic analysis of Pattern B (300 mg scale-up) was done by balancing the sample at 25°C and 50% RH to simulate ambient laboratory conditions. The humidity was then dropped to 0% RH, increased from 0 to 95% RH, dropped from 95 to 0% RH, increased from 0 to 95% RH, and then dropped from 95 to 50% RH. Each point represents the estimated asymptotic weight at each humidity or weight. Pattern B was found to be non-hygroscopic, indicating that the weight loss may be due to the release of residual THF ( Fig.29 ). The main weight loss at the beginning was due to the loss of solvent. XRPD analysis of the sample after moisture sorption analysis was found to be consistent with the described pattern A.

[0334] Pattern D (2-MeTHF solvate, Form V)

[0335] Pattern D (2-MeTHF solvate, Form V) was observed during evaporative crystallization in 2-MeTHF (Examples 4 and 5). XRPD analysis of the solid revealed that a unique crystalline form was obtained, referred to as Pattern D ( Fig.30 ).

[0336] Thermal analysis of Mode D obtained in Example 5 (slow cooling) by DSC showed four endothermic peaks at 67.2°C, 92.2°C, 132.6°C and 150.6°C, followed by degradation after 220°C ( Fig.31 ).

[0337] TGA analysis of the slow cooling amplification of Example 5 showed a 2.7% weight loss between 40-60°C, followed by a 5.3% weight loss between 60-115°C, likely due to the loss of 2-MeTHF followed by decomposition ( Fig.32 ).

[0338] Mode D (slow cooling and enlargement in Example 5) 1 Further HNMR analysis showed that the material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and contained 6.1 wt% residual 2-MeTHF, see Fig.43 .

[0339] Pattern F (2-MeTHF solvate, Form VI)

[0340] Pattern F (2-MeTHF solvate, Form VI) was observed during the 2-MeTHF single solvent crystallization scale-up experiment of 300 mg (Example 6). From the XRPD analysis of the solid, it was found that a unique crystalline form was obtained, which is called Pattern F ( Fig.33 The birefringence observed by optical microscopy confirmed the crystallinity observed by XRPD. Fig.34As shown, the morphology of the crystals was determined to be flake-like with certain aggregation.

[0341] Thermal analysis of Mode F (obtained from Example 6) by DSC showed three endothermic peaks at 93.2°C, 135.2°C and 151.0°C, followed by degradation after 220°C ( Fig.35 ).

[0342] TGA analysis of Pattern F (from Example 6) showed a 1.1% weight loss between 30-110°C, followed by a 0.2% weight loss from 110-160°C, probably due to the loss of 2-MeTHF followed by decomposition ( Fig.36 ). The water content was confirmed by Karl Fischer analysis, which showed that the material contained about 0.1 wt. % water.

[0343] Mode F (obtained from Example 6) 1 Further analysis by H NMR showed that the material was consistent with the structure of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and contained 3.9 wt% residual 2-MeTHF, see Fig.45 .

[0344] Hygroscopic analysis of Pattern F (obtained from Example 6) was performed by equilibrating the sample at 25°C and 50% RH to simulate ambient laboratory conditions. The humidity was then dropped to 0% RH, increased from 0 to 95% RH, dropped from 95 to 0% RH, increased from 0 to 95% RH, and then dropped from 95 to 50% RH. Each point represents the estimated asymptotic weight at each humidity or weight. Pattern F was found to be non-hygroscopic, indicating that the weight loss may be due to the release of residual 2-MeTHF ( Fig.37 The initial major weight loss was due to loss of solvent. The XRPD analysis of the sample after moisture sorption analysis was found to be consistent with the pattern E ( Fig.15 ).

[0345] Example 12. Screening of compounds of the present invention from human skin fibroblasts from patients with Friedreich's Ataxia

[0346] Initial screening was performed to identify compounds that were effective in ameliorating redox disorders. The ability of the test samples to rescue FRDA fibroblasts was tested by the addition of L-buthionine-(S,R)-sulfoximide (BSO) as described in Jauslin et al., Hum. Mol. Genet. 11(24):3055 (2002), Jauslin et al., FASEB J. 17:1972-4 (2003), and International Patent Application WO 2004 / 003565. It has been shown that human dermal fibroblasts from patients with Friedreich's ataxia are highly sensitive to inhibition of de novo glutathione (GSH) synthesis with L-buthionine-(S,R)-sulfoximide (BSO), a specific inhibitor of GSH synthase (Jauslin et al., Hum. Mol. Genet. 11(24):3055 (2002)).

[0347] MEM (amino acid and vitamin-rich medium, catalog number 1-31F24-I) and medium 199 (M199, catalog number 1-21F22-I) with Earle's Balanced Salts but without phenol red were purchased from Bioconcept. Fetal bovine serum was obtained from PAA Laboratories. Basic fibroblast growth factor and epidermal growth factor were purchased from PeproTech. Penicillin-streptomycin-glutamine mixture, L-buthionine-(S,R)-sulfoximide and bovine insulin were purchased from Sigma. Calcein AM was purchased from Anaspec. Cell culture medium was made by combining 125 ml M199 EBS, 50 ml fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin, 2 mM glutamine, 10 μg / ml insulin, 10 ng / ml EGF, and 10 ng / ml bFGF; MEM EBS was added to make the volume 500 ml. This solution was stored at +4°C during the described experiments. Cells were obtained from Coriell Cell Repositories (Camden, NJ; library number GM04078) and grown in 10 cm tissue culture plates. Every two days, they were split at a ratio of 1:3.

[0348] Test samples were provided in 1.5 ml glass vials. Compounds were diluted with DMSO, ethanol or PBS to produce 5 mM stock solutions. Once dissolved, they were stored at -20°C.

[0349] Test samples were screened according to the following protocol:

[0350] Cultures with FRDA fibroblasts were started from 1 ml vials stored in liquid nitrogen with approximately 500,000 cells. The cells were propagated in 10 cm cell culture dishes at a ratio of 1:3 split every two days until 9 plates were available. Once fused, the fibroblasts were harvested. For 54 microtiter plates (96-well-MTP), a total of 14.3 million cells (generation 8) were resuspended in 480 ml of culture medium, equivalent to 100 microliters of culture medium with 3,000 cells / well. The remaining cells were distributed in 10 cm cell culture plates (500,000 cells / plate) for propagation. The plates were incubated overnight at 37°C, 95% humidity and 5% CO2 in an atmosphere to allow the cells to attach to the culture plates.

[0351] 10% DMSO (242.5 microliters) was added to the wells of the microtiter plate. The test compound was thawed and 7.5 microliters of the 5 mM stock solution was dissolved in the wells containing 242.5 microliters of 10% DMSO to obtain a 150 micromolar master solution. Serial dilutions of the master solution were prepared. The time between individual dilution steps was kept as short as possible (usually less than 30 seconds). At least 4 hours after attachment to the MTP, the cells were then treated with various compound dilutions.

[0352] The well plates were kept overnight in a cell culture incubator. The next day, a solution containing BSO was added to the wells in a manner similar to that described in Jauslin et al., Hum. Mol. Genet. 11(24):3055 (2002), Jauslin et al., FASEB J. 17:1972-4 (2003), and International Patent Application WO 2004 / 003565. After 48 hours, three well plates were examined under a phase contrast microscope to verify that cells in the negative control (wells E1-H1) were clearly dead. The culture medium was discarded from all plates and the remaining liquid was removed by gently inverting the plates onto a paper towel. The plates were washed twice with 100 uL of PBS containing calcium and magnesium.

[0353] Then 100 microliters of PBS+Ca+Mg containing 1.2 micromoles of Calcein AM was added to each well. The plate was incubated at 37°C for 30 minutes. Afterwards, the time fluorescence was read on a Gemini fluorescence microplate reader (excitation / emission wavelengths of 485nm and 525nm, respectively). The data was imported into Microsoft Excel (EXCEL is a registered trademark of Microsoft's electronic spreadsheet program) and the EC50 concentration of each compound was calculated using ExcelFit.

[0354] The compounds were tested three times, ie the experiment was performed three times and the passage number of cells was increased by one with each repetition.

[0355] Even at the highest concentration tested (1%), the solvents (DMSO, ethanol, PBS) had neither a negative effect on the viability of non-BSO treated cells nor a beneficial effect on BSO treated fibroblasts. None of the compounds showed autofluorescence. The viability of non-BSO treated fibroblasts was set to 100% and the viability of BSO and compound treated cells was calculated relative to this value.

[0356] The following table summarizes the EC50s of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0357] disease EC50 Friedreich's ataxia +++

[0358] +++ means less than 100nM

[0359] Example 13. Screening of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide from fibroblasts from patients with various oxidative stress disorders

[0360] (R)-2-Hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide The assay was performed using a method similar to that described in Example 12, and cells from patients with other oxidative stress disorders were used instead of FRDA cells.

[0361] The following table summarizes the EC50 of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide for various diseases.

[0362]

[0363]

[0364] +++ means less than 100nM; ++ means 100-500nM

[0365] Example 14. Screening of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide for protection against cisplatin-induced ototoxicity

[0366] Immortomouse from transgenic mice TMConditionally immortalized auditory HEI-OC1 cells from long-term cultures of the cochlea (as described in Kalinec, G. et al., Audiol. Nerootol. 2003; 8, 177-189 / ) were maintained in high glucose Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS under permissive conditions at 33°C, 10% CO2. Cells were pretreated overnight with (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, apoptosis was detected by caspase 3 / 7 activity, and 24h later, incubated with 50uM cisplatin. Cells incubated in a separate dilution were the controls.

[0367] The following table summarizes the EC25 of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide.

[0368] disease EC25 Cisplatin-induced ototoxicity in auditory cells +++

[0369] +++ means less than 100 micromolar

[0370] Example 15. Screening of polymorphic and amorphous compositions of the invention from patient fibroblasts

[0371] The polymorphic and amorphous compositions of the present invention are tested using a screening method similar to that in Examples 12-14, and the polymorphic or amorphous form is substituted for (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide. The polymorphic and amorphous compositions of the present invention can also be tested using a screening method similar to that in Examples 12-14, and the FRDA cells or other cell lines are appropriately replaced with cells obtained from patients suffering from the oxidative stress disorders of the present invention (e.g., MERRF, MELAS, KSS, Alzheimer's disease, pervasive developmental disorders (such as autism), etc.). These compositions are tested for their ability to rescue human dermal fibroblasts from patients with oxidative stress or to protect cells from cisplatin-induced toxicity.

[0372] Example 16. Administration of the composition of the present invention

[0373] The composition of the present invention is a capsule containing 300 mg of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide and a pharmaceutically acceptable carrier. The capsule is taken orally once a day.

[0374] The disclosures of all publications, patents, patent applications, and published patent applications cited herein by identification are hereby incorporated by reference in their entirety.

[0375] Although the foregoing invention has been described in detail by way of example and example for the purpose of clear understanding, it is obvious to those skilled in the art that some minor changes and modifications may be made to the present invention. Therefore, the description and examples should not be interpreted as limiting the scope of the present invention.

Claims

1. A polymorph of an anhydrate or hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is selected from the group consisting of Form II and Form III, wherein the Form II polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 10.85, 11.33, 13.47, 17.37 and 19.33; and The X-ray powder diffraction pattern of the Form III polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.81, 13.74, 14.02, 15.23, 21.10 and 22.

69.

2. The polymorph according to claim 1, wherein the polymorph is Form III.

3. The polymorph of claim 1, wherein the polymorph is Form III, and wherein the angular position can vary by ±0.

1.

4. A composition comprising the polymorph of any one of claims 2-3, wherein the composition contains less than 5% of polymorphs I, II, IV, V, and VI, wherein the Form I polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72; wherein the X-ray powder diffraction pattern of the Form IV polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66; wherein the X-ray powder diffraction pattern of the Form V polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05; and The X-ray powder diffraction pattern of the Form VI polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 15.

71.

5. A composition comprising the polymorph of any one of claims 2-3, wherein at least 95 mol% of the composition is the Form III polymorph, excluding any solvent, carrier or excipient.

6. The polymorph of claim 1, wherein the polymorph is Form II.

7. The polymorph of claim 1, wherein the polymorph is Form II, and wherein the angular position can vary by ±0.

1.

8. A composition comprising the polymorph of any one of claims 6-7, wherein the composition contains less than 5% of polymorphs I and III-VI, wherein the Form I polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72; wherein the X-ray powder diffraction pattern of the Form IV polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66; wherein the X-ray powder diffraction pattern of the Form V polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05; and The X-ray powder diffraction pattern of the Form VI polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 15.

71.

9. A composition comprising the polymorph of any one of claims 6-7, wherein at least 95 mol% of the composition is the Form II polymorph, excluding any solvent, carrier or excipient.

10. A pharmaceutical composition comprising the polymorph according to any one of claims 1 to 3, 6 and 7 or the composition according to any one of claims 4, 5, 8 and 9, and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition has an anhydrate or hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a HPLC purity greater than 95%, excluding any solvent, carrier or excipient.

12. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition has an anhydrate or hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a HPLC purity greater than 99%, excluding any solvent, carrier or excipient.

13. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition has an anhydrate or hydrate of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a HPLC purity greater than 99.9%, excluding any solvent, carrier or excipient.

14. Use of a therapeutically effective amount or an effective amount of the polymorph of any one of claims 1-3, 6 and 7, the composition of any one of claims 4, 5, 8 and 9, or the pharmaceutical composition of any one of claims 10-13 in the preparation of a medicament for treating or inhibiting oxidative stress disorders, regulating one or more energy biomarkers, normalizing one or more energy biomarkers, or enhancing one or more energy biomarkers.

15. The method of claim 14, wherein the medicament is used to treat or inhibit oxidative stress disorder, wherein the oxidative stress disorder is selected from the group consisting of Alpers disease; Barth syndrome; beta oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive external ophthalmoplegia (CPEO); CPT I deficiency; CPT II deficiency; Friedreich's ataxia (FA); glutaric aciduria type II; Kearns-Sayre syndrome (KSS); lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency (LCAD); LCHAD; Leigh syndrome (subacute necrotizing encephalopathy, Leigh Syndrome; Leigh-like syndrome; Leber's hereditary optic neuropathy (LHON); fatal infantile cardiomyopathy (LIC); Luft disease; multiple acyl-CoA dehydrogenase deficiency (MAD); medium-chain acyl-CoA dehydrogenase deficiency (MCAD); myoclonic seizures with ragged red fibers (MERRF); mitochondrial recessive ataxia syndrome (MIRAS); mitochondrial cytopathy, mitochondrial DNA depletion; mitochondrial encephalopathy; mitochondrial myopathy; mitochondrial neurogastrointestinal encephalopathy (MNGIE); neuropathy, ataxia, and retinitis pigmentosa (NARP); Pearson syndrome; pyruvate carboxylase deficiency; pyruvate dehydrogenase deficiency; Respiratory chain disorders; Short-chain acyl-CoA dehydrogenase deficiency (SCAD); SCHAD; Very long-chain acyl-CoA dehydrogenase deficiency (VLCAD); Cardiomyopathy; Encephalopathy; Parkinson's disease; Alzheimer's disease; Amyotrophic lateral sclerosis (ALS); Epilepsy; Macular degeneration; Diabetes; Metabolic syndrome; Brain cancer; Huntington's disease; Schizophrenia; Bipolar disorder; Autism; Asperger's syndrome; Childhood disintegrative disorder (CDD); Rett's disorder; PDD-NOS; Stroke; Optic neuropathy; Dominant juvenile optic atrophy; Toxic drug-induced optic neuropathy; Glaucoma; Stargardt's macular dystrophy maculardystrophy); diabetic retinopathy; diabetic maculopathy; retinopathy of prematurity; ischemia-reperfusion-related retinal damage; oxygen toxicity; thalassemia; sickle cell anemia; epileptic seizures; ischemia; renal tubular acidosis; The group consists of attention-deficit / hyperactivity disorder (ADHD); neurodegenerative diseases causing hearing or balance disorders; dominant optic atrophy (DOA); maternally inherited diabetes and deafness (MIDD); chronic fatigue; contrast-induced renal injury; contrast-induced retinopathy injury; abetalipoproteinemia; retinal atrophy; Wolfram disease; Tourette syndrome; cobalamin C deficiency; methylmalonic aciduria; glioblastoma; Down syndrome; acute tubular necrosis; leukodystrophy; progressive supranuclear palsy; spinal muscular atrophy; noise-induced hearing loss; traumatic brain injury; multiple sclerosis; NGLY1; multiple system atrophy; adrenoleukodystrophy; and adrenoleuroneuropathy.

16. Use according to claim 15, wherein the oxidative stress disorder is multiple system atrophy.

17. The use according to claim 14, wherein the oxidative stress disorder is cancer; mitochondrial myopathy, encephalopathy, lactic acidosis, stroke-like episodes (MELAS); or juvenile Huntington's disease.

18. The use according to claim 15, wherein the oxidative stress disorder is bipolar disorder.

19. The use according to claim 15, wherein the oxidative stress disorder is schizophrenia.

20. The use according to claim 15, wherein the oxidative stress disorder is an age-related disease.

21. The use according to claim 15, wherein the oxidative stress disorder is Huntington's disease.

22. The use according to claim 15, wherein the oxidative stress disorder is Alzheimer's disease.

23. The use according to claim 15, wherein the oxidative stress disorder is amyotrophic lateral sclerosis (ALS).

24. The use according to claim 15, wherein the oxidative stress disorder is epilepsy.

25. The use according to claim 15, wherein the oxidative stress disorder is Parkinson's disease.

26. The use according to claim 15, wherein the oxidative stress disorder is epileptic seizures.

27. The use according to claim 15, wherein the oxidative stress disorder is stroke.

28. A method for preparing a polymorph of anhydrous (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide, wherein the polymorph is Form II, The method comprises the steps of: (a) dissolving (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in EtOAc, (b) rapidly cooling the mixture obtained in step (a), and (c) isolating the (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide; and The X-ray powder diffraction pattern of the Form II polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 10.85, 11.33, 13.47, 17.37 and 19.

33.

29. The method of claim 28, wherein the starting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in step (a) is a Type I polymorph; and The X-ray powder diffraction pattern of the Form I polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.

72.

30. A method for preparing a polymorph of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide hydrate, wherein the polymorph is type III, The method comprises the steps of: (a) mixing (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with an aqueous solution of 0.5% MC / 2% Tween 80 to produce a slurry; (b) slurrying the mixture obtained in step (a); and (c) removing the aqueous solution of 0.5% MC / 2% Tween 80; and The X-ray powder diffraction pattern of the Form III polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.81, 13.74, 14.02, 15.23, 21.10 and 22.

69.

31. The process of claim 30, wherein the starting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in step (a) is a Type I polymorph; and The X-ray powder diffraction pattern of the Form I polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.

72.

32. The method of claim 30 or 31, wherein step (b) is performed for at least 24 hours.

33. A method for preparing a polymorph of (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide hydrate, wherein the polymorph is type III, The method comprises the steps of: (a) mixing (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide with a 0.5% MC aqueous solution to produce a slurry; (b) slurrying the mixture obtained in step (a) for at least 7 days; and (c) removing the 0.5% MC aqueous solution; and The X-ray powder diffraction pattern of the Form III polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.16, 11.81, 13.74, 14.02, 15.23, 21.10 and 22.

69.

34. The method according to claim 33, wherein the starting (R)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxacyclohexa-1,4-dienyl)butanamide in step (a) is polymorphic form I, II, IV, V or VI, respectively; wherein the Form I polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72; The X-ray powder diffraction pattern of the Form II polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.63, 10.85, 11.33, 13.47, and 19.33; wherein the X-ray powder diffraction pattern of the Form IV polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66; wherein the X-ray powder diffraction pattern of the Form V polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05; and The X-ray powder diffraction pattern of the Form VI polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 15.

71.

35. The polymorph of claim 1, wherein the polymorph is Form III having a differential scanning calorimetry (DSC) thermogram comprising an endothermic peak as shown in FIG.

22.

36. A composition comprising the polymorph of claim 35, wherein the composition contains less than 5% of polymorphs I-II and IV-VI; wherein the Form I polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72; wherein the X-ray powder diffraction pattern of the Form IV polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66; wherein the X-ray powder diffraction pattern of the Form V polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05; and The X-ray powder diffraction pattern of the Form VI polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 15.

71.

37. A composition comprising the polymorph of claim 35, wherein at least 95 mole % of the composition is the Form III polymorph, not counting any solvent, carrier, or excipient.

38. The polymorph of claim 1, wherein the polymorph is Form II having a differential scanning calorimetry (DSC) thermogram comprising two endothermic peaks as shown in FIG.

17.

39. A composition comprising the polymorph of claim 38, wherein the composition contains less than 5% of polymorphs Forms I and III-VI; wherein the Form I polymorph has an X-ray powder diffraction pattern comprising characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 12.06, 15.33, 17.03, 17.26, and 18.72; wherein the X-ray powder diffraction pattern of the Form IV polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 4.31, 8.76, 12.97, 13.20, and 16.66; wherein the X-ray powder diffraction pattern of the Form V polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 9.61, 11.49, 12.93, 15.45, and 26.05; and The X-ray powder diffraction pattern of the Form VI polymorph comprises characteristic peaks at at least the following angular positions, wherein the angular positions may vary by ±0.2: 6.27, 9.41, 9.91, 12.94, and 15.

71.

40. A composition comprising the polymorph of claim 38, wherein at least 95 mole % of the composition is the Form II polymorph, not counting any solvent, carrier, or excipient.

41. A composition comprising the polymorph of claim 1, wherein the composition comprises Form II and Form III.

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