A metabolic solution for seizure disorders and traumatic brain injury with antiseizure, neuroprotective, and cardioprotective properties

Abstract

Provided herein are methods and pharmaceutical compositions for preventing or reducing frequency or severity of convulsions or seizures, particularly epileptic seizures by administration of a therapeutically effective amount of 2-deoxy-D-glucose (2-DG) in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones. Also provided are methods and pharmaceutical compositions for reducing severity and progression of traumatic brain injury and deleterious sequelae resulting therefrom in an animal by administration of a therapeutically effective amount of 2-DG in combination with a therapeutically effective amount of beta-hydroxybutyrate, or 2-DG with other ketones or metabolic intermediates that increase brain ketones.

Description

P240371US0124- 1044- WOA METABOLIC SOLUTION FOR SEIZURE DISORDERS AND TRAUMATIC BRAIN INJURY WITH ANTISEIZURE, NEUROPROTECTIVE, AND CARDIOPROTECTIVE PROPERTIESBACKGROUND

[0001] Functions of the central nervous system can be impaired by a variety of paroxysmal alterations including seizures, syncope, pain, migraine, and transient ischemia. The nerve cells of the brain function in a highly complex but organized manner. A sudden temporary interruption in some or all of the functions of the nerve cells results in a “seizure.” Each individual has a “seizure threshold” or level of resistance to seizures: this threshold varies from person to person, most likely due to their genetic makeup and other, developmental factors (Stafstrom, 1998, Pediatrics in Review 19: 335-344).

[0002] A person with a tendency to have repeated seizures may be suffering from epilepsy. Epilepsy is a generic term for a common, serious neurological condition that affects one in every 200 adults and one in every 100 children (Hauser & Hersdorffer, 1990, EPILEPSY: FREQUENCY, CAUSES AND CONSEQUENCES, New York: Demos). Epilepsy is defined by recurrent episodes of seizures, which are brief involuntary behavioral alterations caused by paroxysmal intense electrical discharges in the brain. The causes of epilepsy are heterogeneous and include a diverse variety of genetic, metabolic, developmental, traumatic, neoplastic, and vascular etiologies which may present at any time from birth to senescence.

[0003] As yet there is no known cure for epilepsy and therapeutic intervention is directed at reducing seizure frequency and severity. Currently available antiseizure medications used for chronic epilepsy produce favorable effects in about 7 of 10 people, but complete seizure control, i.e., suppression of all seizures (which is the goal of therapy), occurs in only ~ 50% of patients. Of the remaining 50%, ~ 35% achieve partial suppression with occasional seizures, and ~ 15% have minimal response with frequent, sometimes daily, seizures. Currently available antiseizure medications for acute treatment of continuing seizures referred to as status epilepticus seek to achieve seizure suppression within the recommended 5-minute timeframe to prevent seizure- induced brain damage but have proven to be capable of suppressing the continuing seizures in only a subset of patients.

[0004] A methodology known in the art that is alternative to pharmaceutical intervention involves placing the patient on a diet, termed the ketogenic diet, that has been used in children with epilepsy who have not adequately responded to medical therapy with conventional anticonvulsants (Wilder, 1921 , Mayo Clinic Proceedings 2: 307-308; Freeman et al., 1998, Pediatrics 102: 1358-1363). The anticonvulsant action of the diet, which derives calories fromP240371US0124- 1044- WO high fat intake with very low or no carbohydrates and only adequate protein for growth, is associated with ketosis and production of the ketones beta-hydroxybutyrate and acetoacetate. The ketogenic diet can be significantly efficacious and reduce seizures in a substantial subset of patients with severe epilepsy, but understanding of how the diet produces anticonvulsant effects has been limited. Although the diet induces ketosis and generates ketone bodies (inter alia, beta- hydroxybutyrate, acetoacetate, and acetone), in experimental models ketone bodies are not consistently correlated with the anticonvulsant or anti-epileptic effects; see, Lutas and Yellen, 2013, “The ketogenic diet: metabolic influences on brain excitability and epilepsy’’. Trends Neurosci. 36(1):32-40; Simeone et al., 2018, Do ketone bodies mediate the anti-seizure effects of the ketogenic diet?” Neuropharmacology 133: 233-241).

[0005] Despite its general efficacy, treating patients with the ketogenic diet, particularly children, has several drawbacks. Initiation of the diet typically requires hospitalization for up to one week, and the effects and benefits of the diet (i.e., seizure reduction) are usually not experienced immediately, being delayed from one week to three months from when the diet is started. Maintenance of the diet is difficult, because it requires a balance of nutrients at a particular ratio (usually 3:1 to 4:1 fats to all other nutrients) and intake of even a minimal amount of carbohydrates can eliminate the seizure-relieving benefits of the diet. Side-effects of the diet itself include nausea, vomiting, constipation, depression, sleepiness, lethargy, crankiness, decreased alertness, kidney stones, weight gain, increased serum cholesterol, and acidosis (Ballaban-Gil et al., 1998, Epilepsia 39: 744-748). In addition, the diet has limited effectiveness in adults, and can be even more difficult to implement with children who are allergic to dairy products.

[0006] .Another metabolism-based approach that mimics the effects of the ketogenic diet involves administration of a non-metabolized glucose analog, 2-deoxyglucose (2-DG) as set forth in U.S. Patent Nos. 7,795,227 and 8,633,165, incorporated by reference in their entireties herein. Chronic administration of therapeutically effective dosages of 2-DG have been found to have deleterious effects on heart function, however, which has limited extensive use of this compound to treating acute epileptic seizures.

[0007] Thus there remains a need in this art for methods and pharmaceutical compositions that can more reliably provide treatment to patients with seizure disorders and particularly epilepsy.

[0008] Traumatic Brain Injury (TBI) is a common acquired disorder of the nervous system with a broad spectrum of severity and heterogeneity ranging from lethal penetrating injuries toP240371US0124- 1044- WO closed head injury with concussion. It is estimated that approximately 1 .4 million people in the U.S. experience TBI every year caused by transportation and vehicular accidents, falls, sports injuries, gunshot wounds, and child abuse. Of these, at least 1 million are treated in emergency rooms, about 50,000 people die annually from TBI, and about 230,000 are hospitalized and survive. An additional source of TBI is related to military action abroad due to the frequency of blast injuries, which are increasingly common but survivable due to improvements in acute emergency care in contemporary combat zones. While survivability has increased, individuals who survive TBI are often left with significant cognitive and communicative disabilities, behavioral disorders such as post-traumatic stress disorder (PTSD), and long-term medical complications such as post-traumatic epilepsy (PTE).

[0009] The emergence of PTE and PTSD after TBI is a prototypical example of an acquired brain injury leading to adverse long-term functional consequences. PTE and PTSD may develop and progress despite long intervals after the initial injury, implying that neuronal and brain circuit plasticity initiated by the injury may contribute to development of these disorders. Neural plasticity is the capacity of neurons and neural circuits in the brain to undergo structural and functional modification in response to experience, activity, and injury. While the adult brain was once regarded as “hard-wired” with only limited capacity for adaptation, alteration, and reorganization of function, plasticity is now recognized as a fundamental property of the brain that plays a role not only in development but in learning, memory, cognition, pathological processes, and recovery of function after brain injury. Plasticity is defined as the ability of the brain to undergo changes in structure and function. Cellular processes underlying plasticity are now thought to operate at every level of biological organization in the brain, including molecular and cellular levels as well as circuits, networks, and systems. In regard to TBI, plasticity has been implicated as a potential influence on recovery of function after damage, but in addition, processes of plasticity are also hypothesized to contribute to long-term adverse consequences such as PTE, specifically during the latent period from initial injury to emergence of symptomatic seizures. Development of PTE, PTSD or other deleterious sequelae of TBI is unpredictable, even in individuals who experience TBI of apparent comparable severity and location.

[0010] Currently, TBI therapy is limited primarily to surgical treatment of the initial injury when possible and supportive general medical care. There have been long-standing and continuing efforts to develop new therapies for TBI survivors, with the goal of reducing the initial extent and progression of TBI, and preventing its long-term complications such as PTSD and PTE. Unfortunately, no efficacious therapies for TBI have been demonstrated in the art.P240371US0124- 1044- WOHowever, a wide variety of agents have been evaluated in experimental models of TBI and in clinical trials in TBI patients, comprising more than 38 clinical trials that are underway since 2020 for TBI which include studies of both marketed drugs and new chemical entities in preclinical development.

[0011] Thus, there is a need in the art to develop methods and compounds for treating TBI and its consequences.SUMMARY

[0012] This disclosure provides compositions and methods for preventing or reducing frequency or severity of convulsions or seizures, particularly epileptic seizures by administration of 2-DG in combination with beta-hydroxybutyrate or 2-DG in combination with ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones in therapeutically effective amounts. This disclosure also provides methods and pharmaceutical compositions for reducing incidence, frequency, duration or severity of deleterious sequelae of traumatic brain injury in an animal by administration of 2-DG with beta- hydroxybutyrate or 2-DG in combination with other ketones or metabolic intermediates that increase brain ketones in therapeutically effective amounts. In addition to treatment of seizure disorders these aspects of the methods of the invention are also advantageously applied to a human having a traumatic brain injury, in non-limiting examples acquired by transportation and vehicular accidents, falls, sports injuries, gunshot wounds, child abuse or military action.

[0013] In a first aspect disclosed herein are methods for preventing or reducing the frequency or severity of a convulsion or seizure, particularly epileptic seizure in adult or juvenile animal, the method comprising the step of administering to the animal a therapeutically effective amount each of 2-DG and beta-hydroxybutyrate or 2-DG in combination with such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones. In particular embodiments, the therapeutically effective amount of 2-DG administered to a patient would produce a brain concentration of 2-DG of 2.5-10mM. In particular embodiments the therapeutically effective amount of beta-hydroxybutyrate would produce a brain concentration of 15-25mM.

[0014] Also disclosed in this aspect are methods for reducing incidence, frequency, duration or severity of deleterious sequelae of traumatic brain injury in an animal, the method comprising the step of administering to the animal a therapeutically effective amount each of 2-DG and beta- hydroxybutyrate or 2-DG in combination with other ketones such as acetone, ketone precursors,P240371US0124- 1044- WO metabolic precursors, intermediates, or supplements that increase brain ketones. In particular embodiments, the therapeutically effective amount of 2-DG administered to a patient would produce a brain concentration of 2.5-1 OmM. In particular embodiments the therapeutically effective amount of beta-hydroxybutyrate would produce a brain concentration of 15-25mM.

[0015] In a second aspect provided herein are pharmaceutical compositions for preventing or reducing the frequency or severity of a convulsion or epileptic seizure in an adult or juvenile animal comprising a therapeutically effective amount each of 2-DG and beta-hydroxybutyrate or 2-DG in combination with other ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones and a pharmaceutical ly-acceptable excipient. In particular embodiments the pharmaceutical composition comprises a therapeutically effective amount of 2-DG which produces a brain concentration of 2.5-10mM is administered to a patient. In particular embodiments the therapeutically effective amount of beta-hydroxybutyrate administered to the patient produces a brain concentration of 15-25mM.

[0016] Also disclosed in this aspect are pharmaceutical compositions for reducing incidence, frequency, duration or severity of deleterious sequelae of traumatic brain injury in an animal, the method comprising the step of administering to the animal a therapeutically effective amount each of 2-DG and beta-hydroxybutyrate or 2-DG in combination with other ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones. In particular embodiments, the therapeutically effective amount of 2-DG administered to a patient is 2.5-10mM. In particular embodiments the therapeutically effective amount of beta- hydroxybutyrate is 15-25mM.

[0017] The methods and compositions provided herein can advantageously be used to treat or prevent or reduce the frequency or severity of convulsions or seizure, particularly epileptic seizures that are acute convulsion or epileptic seizures. In certain embodiments, the methods and pharmaceutical compositions are employed wherein the therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered during the convulsion or seizure, particularly epileptic seizure or thereafter. In alternative embodiments the methods and pharmaceutical compositions disclosed herein are administered chronically to an adult or juvenile animal in need thereof to suppress or prevent or reduce the frequency or severity of a convulsion or epileptic seizure. In certain embodiments the therapeutically effective amount of 2-DG and beta-hydroxybutyrate are administered by intravenous injection, preferably by a solution composed of a mixture of 2-DG and beta-P240371US0124- 1044- WO hydroxybutyrate, but alternatively by sequential IV-rapid bolus injection of 2-DG immediately followed by rapid bolus IV injection of beta-hydroxybutyrate.

[0018] The methods and compositions provided herein can advantageously be used to reduce incidence, frequency, duration or severity of deleterious sequelae of traumatic brain injury in an animal. In certain embodiments, the methods and pharmaceutical compositions are employed wherein the therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2- DG in combination with ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered to an animal having a traumatic brain injury. In certain embodiments the therapeutically effective amount of 2-DG and beta-hydroxybutyrate are administered by intravenous injection, preferably by a solution composed of a mixture of 2-DG and beta-hydroxybutyrate, but also by sequential IV rapid bolus injection of 2-DG immediately followed by rapid bolus IV injection of beta-hydroxybutyrate.

[0019] Diseases and disorders treated using the methods and pharmaceutical compositions disclosed herein include but are not limited to epilepsy, migraine, syncope, bipolar disorder, psychosis, anxiety, a stress-inducing disorder, convulsions or a neuropsychiatric disorder having paroxysmal or periodic features. In particular the methods and pharmaceutical compositions disclosed herein can be used to treat status epilepticus. In particular embodiments convulsions treated using the methods and pharmaceutical compositions disclosed herein are associated with epilepsy or epileptic seizures.

[0020] The traumatic brain injuries advantageously treated using the methods and pharmaceutical compositions provided herein include, in non-limiting examples acquired by transportation and vehicular accidents, falls, sports injuries, gunshot wounds, child abuse or military action.

[0021] In particularly advantageous embodiments of the methods and pharmaceutical compositions provided herein the combinations of 2-DG and beta-hydroxybutyrate or 2-DG in combination with ketones such as acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered in sufficient and relative quantities together or in sequence to reduce or prevent cardiac-related deleterious side effects associated with administration of therapeutically effective amounts of 2-DG in the absence or insufficient amounts of beta-hydroxybutyrate.

[0022] These and other features, objects, and advantages of this invention will become better understood from the description that follows. In the description, reference is made to theP240371US0124- 1044- WO accompanying drawings, which form a part hereof and in which there is shown by way of illustration, not limitation, embodiments of the invention. The description of preferred embodiments is not intended to limit the invention to cover all modifications, equivalents, and alternatives. Reference should therefore be made to the claims recited herein for interpreting the scope of the invention.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] An understanding of the invention is facilitated by reference to the drawings.

[0024] FIGs. 1A, 1 B, 1C, 1D, 1 E, 1 F, 1G, and 1 H are graphs showing the effect of the addition of various concentrations of 2-DG and beta-hydroxybutyrate (bHB) on spontaneous epileptic burst discharges in the CA3 region of hippocampal slices bathed in artificial cerebrospinal fluid (ACSF) with extracellular potassium ion concentration ([K+]o) increased to 7.5 mM. FIG. 1A and FIG. 1 B demonstrate effects bath application of 2.5 mM 2-DG followed by sequential additions of 0.1-20 mM bHB in slices from Perforant Path Kindling Susceptible (PPKS) rats (FIG. 1A) and Sprague Dawley (SD) rats (Fig.1 B). The insert in FIG. 1A shows an epileptiform burst discharge in CA3 induced by increasing [K+]oto 7.5mM. FIG. 1C and FIG. 1 D demonstrate effects of 5.0 mM 2-DG followed by sequential additions of 0.1-20 mM bHB in slices from PPKS rats (FIG. 1C) and Sprague Dawley rats (FIG. 1D). FIG. 1 E and FIG. 1 F demonstrate the effects of 0.1-20 mM bHB in slices from both PPKS rats and SD rats. The combination of 2.5-5.0mM 2-DG with 20 mM bHB produced greater reduction in burst rates compared to 2.5- 5.0mM 2-DG alone. FIG. 1G compares the burst rate in slices from both PPKS rats and SD rats exposed to 7.5 mM [K+]ofollowed by 0.1- 20 mM bHB. The bars in FIG. 1 H compare the burst rates in slices from PPKS and SD rats exposed to 20mM bHB to the rates in slices exposed to 2.5 mM 2-DG + 20 mM bHB and 5.0 mM 2-DG + 20 mM bHB. The combination of 2-DG +bHB produced greater reduction in burst rates compared to bHB alone.

[0025] FIG. 2A shows representative sections of hematoxylin-eosin stained cardiac muscle from normal male and female SD rats and rats treated for 21 days with 2-DG or 2-DG+bHB administered by intravenous (IV) injection via femoral catheters. Cardiomyocyte vacuolation induced by 2-DG treatment was reduced by 2-DG+bHB. FIG. 2B provides the Grading Scale for examination of the extent of cardiomyocyte vacuolation in sections of cardiac muscle. FIG. 2C provides bar graphs showing mean histological grading scores in males + females (left panel), males (middle panel), and females (right panel), and demonstrated reduction in severity of cardiac vacuolation in sections from rats treated with 2-DG+bHB compared to rats treated with 2- DG alone.P240371US0124- 1044- WO

[0026] FIG. 3A provides the mean ± standard error of the mean (SEM) for the weights of normal male and female rats hearts, and demonstrates that heart weights of normal males are greater than normal females. FIG. 3B-FIG. 3F are scatter plots demonstrating heart weights of normal control rats vs. rats treated with various doses of 2-DG or 2-DG+bHB. Results from combined groups of male and female rats are provided in FIG. 3B. Males and females heart weights are provided separately at different doses in FIG. 3C-FIG. 3F. The horizontal bars in each plot are mean and SEM for each control or treatment group. The plots demonstrate that 2- DG treatment tended to reduce heart weight and 2-DG+bHB tended to restore heart weights toward normal controls.

[0027] FIG. 4A-FIG. 4C shows duration of status epilepticus (SE) induced in awake SD rats by electrical stimulation of the hippocampus and effects of intraperitoneal (IP) treatment with 2- DG (308 mg / kg) + beta-hydroxybutyrate (936 mg / kg) compared to saline. FIG. 4A is EEG recordings from an epidural electrode in a representative saline-treated rat (upper trace) and a rat treated with 2-DG + bHB (lower trace). FIG. 4B is a spectrogram of EEG power in the same saline-treated rat (upper trace) and the rat treated with 2-DG + bHB (lower trace). Treatment onset is indicated by the black dashed vertical line and cessation of SE is indicated by the red dashed vertical lines with asterisks. FIG. 4C shows duration of SE in the 2-DG + bHB treatment group vs. saline controls. Treatment terminated SE in 7 of 8 rats treated with 2-DG+bHB compared to continuing SE observed in 7 of 7 saline controls (duration differences significant, p<0.01).

[0028] FIG. 5A-FIG. 5B are plots of brain lesion volume measured by ex-vivo computerized tomographic (CT) imaging at 23 days after brain injury induced by controlled cortical impact (CCI) in SD rats. FIG. 5A demonstrates that lesion volume was reduced in rats treated with various IP or IV doses of 2-DG+bHB beginning immediately after CCI and continued for 14-21 days compared to untreated controls or rats that received IV or IP injections of saline for matched periods of 14-21 days. FIG. 5B is a plot of lesion volume at 23 days across all examined doses of 2-DG + bHB treated rats for 14-21 days vs. rats matched for treatment with saline for 14-21 days.DETAILED DESCRIPTION

[0029] This disclosure provides methods for treating, by preventing or reducing frequency or severity of convulsions or seizures, particularly those associated with epilepsy, by administration of therapeutically effective amounts of 2-deoxyglucise (2-DG) or glycolysis-inhibiting analogs thereof in combination with beta-hydroxybutyrate, or 2-deoxyglucose (2-DG) or glycolysis-P240371US0124- 1044- WO inhibiting analogs thereof in combination with other ketones or metabolic intermediates that increase brain ketones. Also provided are pharmaceutical compositions of 2-deoxyglucose (2- DG) or glycolysis-inhibiting analogs thereof in combination with beta-hydroxybutyrate or with other ketones or metabolic intermediates that increase brain ketones comprising therapeutically effective amounts of both components of the pharmaceutical compositions.

[0030] This disclosure also provides methods for reducing the severity and deleterious sequelae of TBI, which include progressive neurological dysfunction, progressive structural damage and brain loss (atrophy), post-traumatic epilepsy, and post-traumatic stress disorder, comprising administering to a TBI victim in need thereof a therapeutically effective amounts of 2- deoxyglucose 2-DG or glycolysis-inhibiting analogs thereof in combination with beta- hydroxybutyrate, or 2-DG or glycolysis-inhibiting analogs thereof in combination with other ketones or metabolic intermediates that increase brain ketones. Also provided are pharmaceutical compositions of 2-deoxyglucose (2-DG) or glycolysis-inhibiting analogs thereof in combination with beta-hydroxybutyrate or with other ketones or metabolic intermediates that increase brain ketones comprising therapeutically effective amounts of both components of the pharmaceutical compositions.DEFINITIONS

[0031] Prior to setting forth this disclosure in more detail, it may be helpful to provide definitions of certain terms to be used herein. Additional definitions are set forth throughout this disclosure.

[0032] As used in the specification and claims, the singular form “a,” “an,” and “the” includes plural references unless the context clearly dictates otherwise. It should be understood that the terms “a” and “an” as used herein refer to “one or more” of the enumerated components.

[0033] The use of the alternative (e.g., “or”) should be understood to mean either one, both, or any combination thereof of the alternatives.

[0034] The term “about” as used herein in the context of a number refers to a range centered on that number and spanning 10% less than that number and 10% more than that number. The term “about” used in the context of a range refers to an extended range spanning 15% less than that of the lowest number listed in the range and 10% more than the greatest number listed in the range.

[0035] Throughout this disclosure, any concentration range, percentage range, ratio range,P240371US0124- 1044- WO or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. Also, any number range of this disclosure relating to any physical feature, such as polymer subunits, size, or thickness, are to be understood to include any integer within the recited range, unless otherwise indicated. Throughout this disclosure, numerical ranges are inclusive of their recited endpoints, unless specifically stated otherwise.

[0036] Unless the context requires otherwise, throughout this specification and claims, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” As used herein, the terms “include” and “comprise” are used synonymously.

[0037] The phrase “at least one of” when followed by a list of items or elements refers to an open-ended set of one or more of the elements in the list, which may, but does not necessarily, include more than one of the elements.

[0038] As used herein, the term “paroxysmal disorder” includes but is not limited to seizure disorders such as infantile spasms, myoclonic and “minor motor” seizures, as well as tonic-clonic seizures and partial complex seizures. In preferred embodiments, the seizure disorder is epilepsy, including idiopathic, symptomatic and cryptogenic epilepsy, and more preferably drugresistant or medically-refractory epilepsy, status epilepticus by which is meant that epileptic seizures continue despite adequate administration of antiepileptic drugs.

[0039] Further as used herein, the term “paroxysmal disorders” also includes syncope, convulsive syncope, migraine, pain, tics, tremors and other movement disorders, and neuropsychiatric conditions with paroxysmal or intermittent behavioral disturbances including bipolar disorders, affective disorders, anxiety disorders, and stress disorders.

[0040] As used herein, the term “metabolic precursors” or “metabolic intermediates” includes chemical congeners which are converted to ketones such as ketone esters, intermediates in ketogenesis which are metabolized to ketones, amino acids which are catabolized into ketones, and supplements which elevate brain ketones.

[0041] As used herein, the term “ketone precursors” is intended to encompass, inter alia, free fatty acids, lactate, and molecules of intermediary metabolism which contribute to ketogenesis.

[0042] Other particular embodiments that would be familiar to those skilled in the art include but are not limited to chemical congeners converted to ketones such as ketone esters (e.g. , R,SP240371US0124- 1044- WO1 ,3 butanediol), intermediates in ketogenesis such as 3 - hydroxy- 3 - methylglutaryl CoA (HMGCoA) to be metabolized to ketones, amino acids (such as leucine) catabolized into ketones, and supplements including free fatty acids which may be converted to ketones, and medium chain trigycerides (MCTs) which elevate brain ketones.

[0043] As used herein, an “effective amount” or “therapeutically effective amount” of 2-DG and beta-hydroxybutyrate, or 2-DG in combination with other ketones or metabolic intermediates that increase brain ketones is defined as an amount that when administered to an animal, preferably a human, more preferably a human having a paroxysmal disorder including both adults and juvenile humans with epilepsy, reduces the frequency, duration or severity of seizures experienced by the individual. The “effective amounts” of said compounds are those doses that produce subnanomolar to millimolar concentrations of a compound such in blood or plasma, and will depend on species, pharmacokinetics, and route of administration. In particular, therapeutically effective doses of 2-DG that achieve brain concentrations of 2.5-1 OmM and therapeutically effective doses of beta-hydroxybutyrate other ketones or metabolic intermediates that increase brain ketones achieve brain ketone concentrations of 15-25mM, although lesser doses may also be effective.

[0044] As provided herein, pharmaceutical compositions comprising 2-DG and methods using said compositions will be understood to encompass preparations of 2-DG as the D- stereoisomer, as well as racemic mixtures thereof comprising any combination of D- and L- 2- deoxyglucose, provided that the percentage of the D- stereoisomer is greater than zero. 2-DG is available commercially, and preferably is produced according to the standards and guidelines of the pharmaceutical industry and in compliance with all relevant regulatory requirements. 2-DG can also be synthesized using methods well-established in the art (see, for example, THE MERCK INDE , 12thEd., Monograph 2951 , New Jersey: Merck & Co., 1997; Bergmann et al., 1922, Ber. 55: 158; Snowden et al., 1947, JACS 69: 1048; Bolliger et al., 1954, Helv. Chim. Acta 34: 989; Bolliger, 1962, “2-Deoxy-D-a / -ab / no-hexose (2-Deoxy-d-glucose),” in METHODS IN CARBOHYDRATE CHEMISTRY, vol. I, (Whistler & Wolfram, eds.), New York Academic Press, pp. 186,189). In certain embodiments a combination comprising a therapeutically effective amount of a racemic mixture of R-beta-hydroxybutyrate and D-beta-hydroxybutyrate or a salt thereof is administered in combination with a therapeutically effective amount of 2-DG. In alternative embodiments a combination comprising a therapeutically effective amount of a substantially enantiomerically pure preparation of R- beta-hydroxybutyrate or a salt thereof is administered in combination with a therapeutically effective amount of 2-DG.P240371US0124- 1044- WO

[0045] The invention provides pharmaceutical compositions of combinations of therapeutically effective amounts each of 2-DG and beta-hydroxybutyrate as well as other ketones and metabolic intermediates which increase brain ketones. The pharmaceutical compositions provided herein can be manufactured in a manner that is itself known, e.g., by means of a conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Pharmaceutical compositions provided herein can be formulated and administered through a variety of means, including systemic, localized, or topical administration. Techniques for formulation and administration can be found in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA. The mode of administration can be selected to maximize delivery to a desired target site in the body. In particular embodiments the pharmaceutical compounds provided herein are administered intravenously.

[0046] Pharmaceutical compositions for use in accordance with the methods disclosed herein can be formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the therapeutic compounds into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.

[0047] The compounds comprising pharmaceutical compositions disclosed herein can be formulated for administration by intravenous injection, e.g., by bolus injection or continuous infusion of a solution composed of a mixture of 2-DG and beta-hydroxybutyrate, or by sequential IV rapid bolus injection of 2-DG immediately followed by rapid bolus IV injection of beta- hydroxybutyrate. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and / or dispersing agents. Such pharmaceutical formulations include aqueous solutions of the active compounds in water-soluble form. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient can be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. In certain embodiments the components of the pharmaceutical compositions provided herein can be provided for oral administration, either in combination or as sequential or simultaneous administration.P240371US0124- 1044- WO

[0048] For injection, compounds comprising pharmaceutical compositions disclosed herein can be formulated in appropriate aqueous solutions, such as physiologically compatible buffers such as Hank's solution, Ringer's solution, lactated Ringer’s solution, or physiological saline buffer. Fortransmucosal and transcutaneous administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0049] Pharmaceutical compositions suitable for use as disclosed herein include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose. More specifically, a therapeutically effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0050] For any compounds used in the methods and pharmaceutical compositions disclosed herein, the therapeutically effective dose can be estimated initially from in vitro assays, as disclosed herein, or using art-recognized animal model systems or a combination thereof. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the ECso (effective dose for 50% increase) as determined in vitro, i.e., the concentration of the test compound which achieves a half-maximal amount of seizure frequency. Such information can be used to more accurately determine useful doses in humans.

[0051] It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the compounds employed, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination, the severity and extent of the particular seizure disorder in the patient undergoing therapy and the judgment of the prescribing physician and in particular the age of the patient, who is may be an adult, a juvenile, a child or an infant.

[0052] Preferred compounds provided by the invention will have certain pharmacological properties. Such properties include, but are not limited to bioavailability, low toxicity, low serum protein binding and desirable in vitro and in vivo half-lives. Assays may be used to predict these desirable pharmacological properties. Assays used to predict bioavailability include transport across human intestinal cell monolayers, including Caco-2 cell monolayers. Serum protein binding may be predicted from albumin binding assays. Such assays are described in a review by Oravcova et al. (1996, J. Chromat. B 677: 1-27). In vitro half-lives of antiglycolytic compounds may be predicted from assays of microsomal half-life as described by Kuhnz and Gieschen (1998, Drug Metabolism and Disposition, 26: 1120-1127).P240371US0124- 1044- WO

[0053] Toxicity and therapeutic efficacy of said compounds can be determined by conventional pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD5o (the dose lethal to 50% of the population) and the ED5o (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, and it can be expressed as the ratio between LD5o and ED5o. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED5o with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition. (See, e.g. Fingl et al., 1975, in "The Pharmacological Basis of Therapeutics", Ch.1 , p.1).

[0054] The pharmaceutical compositions disclosed herein can be administered before, during or after the occurrence of a paroxysmal event such as a seizure, particularly an epileptic seizure, and the route of administration and administered dose chosen accordingly. For example, administration of the pharmaceutical compositions of the invention during a seizure will preferably be in a rapidly-bioavailable dosage using a safe and effective administration route (inter alia, which may not include oral formulations in these embodiments).

[0055] The invention provides methods for reducing seizure frequency, duration or intensity in an animal, preferably an adult or juvenile human. The methods of the invention are effective for reducing seizure frequency, duration or intensity in at least 50%, more preferably 60%, more preferably 70%, more preferably 80%, more preferably 90%, more preferably 95%, more preferably 98%, and more preferably 99% of treated patients. In preferred embodiments, the inventive methods are practiced using the pharmaceutical compositions of the invention as disclosed herein.

[0056] The invention also provides methods for reducing the sequelae of TBI including structural damage progression, cognitive dysfunction, posttraumatic epilepsy, and posttraumatic stress disorder.EXAMPLES

[0057] The following examples are intended to illustrate various embodiments of the invention. As such, the specific embodiments discussed are not to be constructed as limitations on the scope of the invention. It will be apparent to one skilled in the art that various equivalents, changes, and modifications may be made without departing from the scope of invention, and it isP240371US0124- 1044- WO understood that such equivalent embodiments are to be included herein. Further, all references cited in the disclosure are hereby incorporated by reference in their entirety, as if fully set forth herein.Example 1Enhanced antiseizure effects of combined 2-DG and bHB on epileptic bursts in rat hippocampus [ 0 0 5 8 ] Horizontal hippocampal slices (-500 pm thickness) were prepared from Perforant Path Kindling Susceptible (PPKS) and Sprague-Dawley (SD) rats aged 90-120 days. PPKS rats are an inbred strain derived from SD rats that have a heritable trait whereby they exhibit a "fast" kindling rate of 10.7 ± 1.1 afterdischarges (ADs) to a first generalized tonic clonic (Class V) in response to stimulation of the perforant path (see, Langberg et al., 2015, Distinct behavioral phenotypes in novel "fast" kindling-susceptible and "slow" kindling-resistant rat strains selected by stimulation of the hippocampal perforant path, Neurobiol. Dis. 85: 122-129). Hippocampal slices from PPKS and SD rats were maintained in artificial cerebrospinal fluid (ACSF) according to published methods (see, Pan et al., 2010, Ictal Activity Induced by Group I Metabotropic Glutamate Receptor Activation and Loss of Afterhyperpolarizations, Neuropharm. 59: 86-92). Briefly, following deep anesthesia with isoflurane, SD and PPKS rats (age 90-120 days) were decapitated; the brain was removed to prepare hippocampal slices (thickness -500 um) using a Lecia vibratome. During slice preparation iced ACSF containing 3.5 mM KCI, elevated magnesium (7 mM MgCh) and low calcium (0.5 mM CaCI2) was used to bathe the brain. Sectioned slices were transferred and incubated in an interface chamber and bathed in ACSF composed of 130mM NaCI, 7.5 mM KCI, 1.25 mM NaH2PO4, 2mM CaCI2, 2 mM MgCI2, 24mM NaHCOs, and 10 mM glucose at 35°C for 90-120 min. Spontaneous extracellular field potentials were recorded in the stratum pyramidale of the CA3 region of the hippocampal slices (see, Cherubini & Miles, 2015, Front. Cell. Neurosci. 9: 19) using a glass pipette filled with 2M NaCI having impedances of 5-10 MQ.

[0059] FIG. 1A (insert) shows a graph of an epileptiform burst discharge in CA3 induced by increasing [K+]oto 7.5 mM. FIG. 1A and FIG. 1 B demonstrate effects bath application of 2.5 mM 2-DG followed by sequential additions of 0.1-20 mM bHB in slices from Perforant Path Kindling Susceptible (PPKS) rats (FIG.1A) and Sprague Dawley (SD) rats (FIG. 1 B). FIG. 1C and FIG. 1D are graphs showing the effect of the addition of 5.0 mM 2-DG followed by sequential additions of 0.1-20 mM bHB in slices from PPKS rats (FIG.1C) and SD rats (FIG. 1 D). In slices from PPKS rats (FIG.1A, 36 slices, n= 3 male, n=3 female), bath addition of 2.5 mM 2-DG alone followed by sequential concentrations of 0.1-20 mM bHB reducedP240371US0124- 1044- WO epileptic burst charges in CA3 (Krusksal-Wallis One Way Analysis of Variance by Ranks, p < 0.001, H = 52.85, df = 5). Bath addition of 5.0 mM 2-DG alone followed by sequential concentrations of 0.1-20 mM bHB (Fig. 1C, 27 slices, n = 3 male, n = 2 female) also reduced epileptic burst charges in CA3 (ANOVA, p < 0.001 , H = 50.09, df = 5). In slices from SD rats (FIG. 1 B, 36 slices, n - 3 male, n - 2 female) bath addition of 2.5 mM 2-DG alone and in combination with sequential concentrations of 0.1-20 mM bHB reduced epileptic burst charges in CA3 (ANOVA, p < 0.001, H = 58.98, df = 5). In slices from SD rats (FIG. 1 D, 36 slices, n =5 male, n=1 female)) bath addition of 5.0 mM 2-DG alone and in combination with specific concentrations of 0.1-20 mM bHB also reduced epileptic burst charges in CA3 (ANOVA, p < 0.001 , H = 74.57, df = 5). In slices from both PPKS and SD rats bath additions of both 2.5 mM and 5.0 mM 2-DG alone induced concentrationdependent decreases in burst rates by 45-52% compared to control rates in 7.5 mM [K+]o(pair-wise comparison (Dunn's method) (see FIG. 1A-FIG. 1 D). Notably, the concentration-dependent decreases with exposure to combined solutions of 2-DG and bHB (~69-76%) were greater than 2-DG alone (45-52%). In FIG. 1E in slices from PPKS rats (35 slices, n = 3 male, n =2 female) sequential addition of bHB alone at concentrations of 0.1 -1.0 mM had no significant effect on burst rates, but with addition of 20mM bHB, burst rates decreased compared to 7.5 mM [K+]oalone (p < 0.01) (Dunn's pair-wise comparison). In FIG.1F slices from SD rats (43 slices , n = 4 males, n = 2 females) burst rates decreased at 10mM concentrations of bHB (p < 0.01), and further decreased with addition of 20mM bHB alone (p < 0.001) compared to 7.5 mM [K+]oalone (Dunn's pair-wise comparison).Bath application of bHB alone induced concentration-dependent decreases in burst rates by 45-65% compared to control rates in 7.5 mM [K+]o(FIG. 1 E and FIG. 1F). Notably, the concentration-dependent decreases with exposure to combined solutions of 2-DG and bHB (-69-76%) exceeded the decreases with exposure to bHB alone (-45-58%) in slices from PPKS and SD rats (FIG.1G). In FIG. 1 H the bars compare the burst rates in slices from PPKS and SD rats exposed to 20mM bHB to the rates in slices exposed to 2.5 mM 2-DG + 20 mM bHB and 5.0 mM 2-DG + 20 mM bHB. Compared to bHB alone, 2.5 mM 2-DG + 20mM bHB and 5.0mM 2-DG +20mM bHB produced greater reduction in burst rates (p< 0.03 ANOVA, p<0.046, p < 0.049, respectively).These results showed that the combination of 2-DG and bHB exhibited additive, dosedependent effects on spontaneous epileptic burst discharges compared to either 2-DG alone (FIG. 1A-FIG. 1 D) or bHB alone in hippocampal slices from PPKS and normal SD rats (FIG. 1 E, FIG. 1F, and FIG. 1G).P240371US0124- 1044- WOExample 2Cardioprotective effects of bHB against 2-DG-induced cardiomyocyte vacuolation

[0060] 2-DG was administered to male and female SD rats by intravenous (IV) femoral catheters for 21 consecutive days in doses of 250-535 mg / kg / day as split twice daily doses separated by ~8-9 hours. Another group received 2-DG (250-535 mg / kg / day) in twice daily split doses + bHB (660-936 mg / kg) with each split dose. Control rats included normal SD rats and SD rats that received twice daily saline for 21 days by IV femoral catheters. The rats were sacrificed immediately after the last dose and the hearts were removed and placed into fixative consisting of an aqueous solution of 10% formalin. After fixation for 48 hours in 10% formalin, the hearts were weighed and bisected into longitudinal blocks from base to apex. Longitudinal blocks containing both atria and ventricles were embedded in paraffin. Paraffin-embedded tissues were sectioned at 10pm and stained with hematoxylin and eosin (H&E) followed by microscopic examination at 10x and higher magnification. Extent of vacuolization was assessed as percentage of the section area showing vacuolization using a scale of 0 for < 1% vacuolization and up to 4 for >41% vacuolization (FIG. 2B). FIG. 2A shows H&E stained sections of cardiac muscle from a normal male SD rat (upper left panel) and representative sections from males treated with 2-DG (250 mg / kg / day) (upper middle left panel), 2-DG (250 mg / kg / day) + bHB (936 mg / kg bid) (upper middle panel), 2-DG (>250 mg / kg / day) (upper middle right panel), and 2-DG (>250 mg / kg / day) + bHB (936 mg / kg bid) (upper right panel). Corresponding representative sections from SD females include a normal female (lower left panel) and representative sections from females treated with 2-DG (250 mg / kg / day) (lower middle left panel) , 2-DG (250 mg / kg) + bHB (936 mg / kg bid) (lower middle panel), 2-DG (>250 mg / kg / day) (lower middle right panel), and 2-DG (>250 mg / kg / day) + bHB (936 mg / kg bid) (lower right panel). Cardiomyocyte vacuolation scores of combined 2-DG treated male (n=8) and female (n=7) rats (mean 3.04 ± 0.24) vs. combined male (n=9) and female (n=11) rats treated with 2- DG+bHB (mean 2.21 ± 0.17) were significantly reduced (p=0.039) (FIG. 2C, left panel). Cardiomyocyte vacuolation was also significantly reduced by 2-DG+bHB in males (3.18 ± 0.32 vs. 2.23 ± 0.61 , p = 0.031) (FIG. 2C middle panel). In female rats there was a trend toward reduction which did not achieve statistical significance (3.42 ± 0.20 vs. 2.73 ± 0.14, p=0.054) (FIG. 2C right panel).P240371US0124- 1044- WOExample 3Effects of bHB against 2-DG-induced changes in heart weight

[0061] The weights of the fixed hearts from the cohorts of male and female SD rats receiving 2-DG, 2-DG + bHB, or saline by intravenous (IV) femoral catheter for 21 consecutive days as described in Example 2 were compared. Heart weights were measured after fixation for 48 hours in 10% formalin. FIG. 3A provides the mean ± standard error of the mean (SEM) for the weights of normal male (n=8) and female (n=8) rat hearts, and demonstrates that heart weights of normal males are greater than normal females. FIG. 3B-FIG. 3F are scatter plots demonstrating heart weights of normal control and saline-injected rats vs. rats treated with various doses of 2-DG or 2-DG+bHB. Results from combined groups of male and female rats are shown in FIG. 3B. Male and female heart weights from control and treated groups are shown separately at different doses in FIG. 3C-FIG. 3F. The horizontal lines in each plot are mean and ±SEM for each control or treatment group. While mean heart weight differences did not achieve statistical significance at the level of p<0.05 across groups, the plots demonstrate that 2-DG treatment tended to reduce heart weight and 2-DG+bHB tended to restore heart weights toward normal controls.Example 4Effects of 2-DG and bHB treatment on status epilepticus induced by electrical stimulation

[0062] Status epilepticus (SE) was induced in awake, freely moving SD rats by electrical stimulation of the left posterior hippocampus with a bipolar electrode (placed at 6mm posterior to bregma, 5mm lateral, 5mm deep). Electrical current consisting of 0.75msec biphasic pulses at 50Hz was delivered at twice the threshold required to evoke an afterdischarge (AD) for 10 seconds followed by 5 seconds without stimulation for a total duration of 60 minutes. Electroencephalographic (EEG) activity was continuously recorded from epidural screw electrodes over the left and right frontal regions. The high frequency stimulation trains administered for 60 minutes reliably induced continuous ictal EEG activity consisting of runs of irregular and synchronous high amplitude spike activity which continued after cessation of stimulation in all rats. At 15 minutes after the end of stimulation, SD rats demonstrating ictal EEG activity were treated with IP injection of 2-DG (305 mg / kg) and bHB (936mg / kg), or an equivalent volume of saline. EEG activity was monitored for 240 minutes after treatment. FIG. 4A is EEG recording from an implanted epidural electrode in aP240371US0124- 1044- WO representative saline-treated rat (upper trace) and a rat treated with 2-DG + bHB (lower trace). FIG. 4B is a spectrogram of EEG power in the same saline-treated rat (upper trace) and the rat treated with 2-DG + bHB (lower trace). The schematic bar above the upper trace in FIG. 4A indicates periods of stimulation, treatment, and duration of EEG recording. Treatment onset is indicated by the black dashed vertical line and cessation of SE is indicated by the red dashed vertical line with asterisks. Treatment with 2-DG + bHB produced sustained termination of SE in 7 of 8 rats vs. continued SE without termination in 7 of 7 saline-treated rats (proportion significantly different, p=0.0035, Chi-square with Yates correction, x2 = 7.292, df= 1 ).

[0063] FIG. 4C is a plot of the duration (minutes) of electrographic SE in individual saline- treated rats vs. rats treated with 2-DG (305 mg / kg) + bHB (936 mg / kg). The horizontal lines are mean ± SEM of duration of SE for the groups. SE persisted for the duration of the recording (~ 240 minutes after induction in saline- treated rats) (n = 7). The duration of SE in the 2-DG + bHB treatment group during the 240 minute period of observation was significantly reduced compared to saline-treated controls (p<0.01).Example 5Effects of 2-DG and 2-DG+ bHB treatment on brain lesion volume after TBI induced by CCI in rats

[0064] The effects of treatment with 2-DG and 2-DG + bHB on brain lesion volume measured by ex-vivo computerized tomographic (CT) imaging were evaluated in SD rats after brain injury induced by controlled-cortical-impact (CCI). CCI was induced in the right hemisphere of isoflurane anesthetized rats by a Leica Impact One Stereotaxic impactor (Leica), utilizing a 3 mm circular blunt impact tip with a velocity of 6 m / sec and a dwell time of 500 ms. Immediately following CCI, rats received IV femoral injections bid for 21 days with saline (n=7), 2-DG (386 mg / kg / day administered as 173 mg / kg bid) (n=2), 2-DG (386 mg / kg / day administered as 173 mg / kg bid) + bHB (936 mg / kg bid) (n=6). Another group received once daily IP injections 2-DG 250 mg / kg / day (n= 12), 2-DG (250 mg / kg / day) + bHB (936 mg / kg) (n=15), or saline (n=6) for 14 days. At 23 days following CCI, the rats were euthanized with CO2, and brains were rapidly removed and placed into 10% formalin. The brains were analyzed in cohorts which included saline-treated controls. The fixed brains were imaged ex-vivo by computerized tomographic (CT) methods with a MILabs CT-UHR (ultra-high resolution) camera in accurate step and shoot ultra-focus mode, with 85 ms exposure time, 50 kVp x-ray tubeP240371US0124- 1044- WO voltage, 0.21 mA x-ray tube current, 0.10 degree step angles and 1x1 binning. Raw data were subsequently reconstructed with 20-micron isotropic voxels. Volumetric image analysis was performed with 3D Slicer (version 5.8.1 , www.slicer.org). Segmentation of the brain tissue was determined by Otsu automatic thresholding of the raw data. The superficial lesion boundary was set by a line drawn from the cortical edges of the injury. Ventricles were excluded from lesion volume even if continuous with the lesion by approximating the ventricle boundary. Treatment with 2-DG (250mg / kg / day), 2-DG (250mg / kg / day) + BHB (936mg / kg / day), 2-DG 380 mg / kg / day, and 2-DG (350 / kg / day) + bHB (1872 mg / kg / day) IP for 14 days reduced lesion size by an average of ~ 50% across treatment groups vs. saline -treated controls. Lesion volume was also reduced by an average of ~ 50% vs. saline controls when all 2-DG, 2-DG + BHB dose and administration regimens were pooled. No differences in lesion volume were observed as a function of dose of 2-DG, dose of 2-DG + BHB, administration route, or with 14 vs. 21 days of 2- DG or 2-DG + bHB treatments (as shown in FIG. 5A and Fig. 5B).

[0065] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, and patent application was specifically and individually indicated to be incorporated by reference

[0066] While some embodiments have been illustrated and described in detail in the appended drawings and the foregoing description, such illustration and description are to be considered illustrative and not restrictive. Other variations to the disclosed embodiments can be understood and effected in practicing the claims, from a study of the drawings, the disclosure, and the appended claims. The mere fact that certain measures or features are recited in mutually different dependent claims does not indicate that the combination of these measures or features cannot be used. Any reference signs in the claims should not be construed as limiting the scope.

Claims

AMENDED CLAIMS received by the International Bureau on 04 April 2026 (04.04.2026)1 . A method for preventing or reducing the frequency or severity of a convulsion or epileptic seizure in an adult or juvenile animal, the method comprising the step of administering to the animal a therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones.

2. The method of claim 1 , wherein beta-hydroxybutyrate is a racemic mixture of R- beta- hydroxybutyrate and D- beta-hydroxybutyrate or a salt thereof3. The method of claim 1 , wherein beta-hydroxybutyrate is a substantially enantiomerically pure preparation of R- beta-hydroxybutyrate or a salt thereof.

4. The method of claim 1 , wherein the therapeutically effective amount of 2-DG achieves a brain concentration of 2.5-1 OmM5. The method of claim 1 , wherein the therapeutically effective amount of beta- hydroxybutyrate achieves a brain concentration of 15-25mM.

6. The method of clam 1 wherein the combination is 2DG and acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones.

7. A pharmaceutical composition for preventing or reducing the frequency or severity of a convulsion or epileptic seizure in an adult or juvenile animal comprising a therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones and a pharmaceutically-acceptable excipient.

8. The pharmaceutical composition of claim 7, wherein beta-hydroxybutyrate is a racemic mixture of R- beta-hydroxybutyrate and D- beta-hydroxybutyrate or a salt thereof.

9. The pharmaceutical composition of claim 7, wherein beta-hydroxybutyrate is a substantially enantiomerically pure preparation of R- beta-hydroxybutyrate or a salt thereof.

10. The pharmaceutical composition of claim 7, wherein the therapeutically effective amount of 2-DG achieves a brain concentration of 2.5-1 OmM.11 . The pharmaceutical composition of claim 7, wherein the therapeutically effective amount of beta-hydroxybutyrate achieves a brain concentration of 15-25mM.2612. The pharmaceutical composition of claim 7, wherein the combination is 2DG and acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones.

13. The method of any one of claims 1 to 6, wherein the convulsion or epileptic seizure is an acute convulsion or epileptic seizure.

14. The pharmaceutical composition of any one of claims 7 to 12 , wherein the convulsion or epileptic seizure is an acute convulsion or epileptic seizure.

15. The method of claim 13, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered during the convulsion or epileptic seizure or thereafter.

16. The pharmaceutical composition of claim 14, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered during the convulsion or epileptic seizure or thereafter.

17. The method of any one of claims 1 to 6, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or with or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered chronically to the adult or juvenile animal in need thereof to suppress or prevent a convulsion or epileptic seizure.

18. The pharmaceutical composition of any one of claims 7 to 12, wherein the composition is formulated wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered chronically to the adult or juvenile animal in need thereof to suppress or prevent a convulsion or epileptic seizure.

19. The pharmaceutical composition of claims 16 or 18, wherein the pharmaceutical composition is formulated for intravenous administration or formulated for oral administration.

20. The method of claims 15 or 17, wherein the therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones is administered intravenously either simultaneously or sequentially.21 . The method of claim 20 wherein the convulsion or epileptic seizure is caused by a disease or disorder that is epilepsy, migraine, syncope, bipolar disorder, psychosis, anxiety, a stress-inducing disorder, convulsions or a neuropsychiatric disorder having paroxysmal or periodic features.

22. The method of claim 21 , wherein the disease or disorder is status epilepticus.

23. The pharmaceutical composition of claim 19, wherein the convulsion or epileptic seizure is caused by a disease or disorder that is epilepsy, migraine, syncope, bipolar disorder, psychosis, anxiety, a stress-inducing disorder, convulsions or a neuropsychiatric disorder having paroxysmal or periodic features.

24. The pharmaceutical composition of claim 23, wherein the disease or disorder is status epilepticus.

25. The method of claim 17, wherein the convulsion is associated with an epileptic seizure.

26. The pharmaceutical composition of claim 18, wherein the convulsion is associated with an epileptic seizure.

27. A method for reducing severity and progression of traumatic brain injury and deleterious sequelae resulting therefrom in an animal, comprising the steps of administering to an animal in need thereof a therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones.

28. The method according to claim 27, wherein the deleterious sequelae are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic stress disorder, or other adverse consequences of the initial traumatic brain injury.

29. The method of claim 27, wherein beta-hydroxybutyrate is a racemic mixture of R- beta- hydroxybutyrate and D- beta-hydroxybutyrate or a salt thereof30. The method of claim 27, wherein beta-hydroxybutyrate is a substantially enantiomerically pure preparation of R- beta-hydroxybutyrate or a salt thereof.31 . The method of claim 27, wherein the therapeutically effective amount of 2-DG achieves a brain concentration of 2.5-1 OmM.

32. The method of claim 27 wherein the combination is 2DG and acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones.

33. The method of claim 27, wherein the therapeutically effective amount of beta- hydroxybutyrate achieves a brain concentration of 15-25mM.

34. A pharmaceutical composition reducing severity and progression of traumatic brain injury and deleterious sequelae resulting therefrom in an animal, comprising a therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones and a pharmaceutically-acceptable excipient.

35. The pharmaceutical composition of claim 34, wherein beta-hydroxybutyrate is a racemic mixture of R- beta-hydroxybutyrate and D- beta-hydroxybutyrate or a salt thereof.

36. The pharmaceutical composition of claim 34, wherein beta-hydroxybutyrate is a substantially enantiomerically pure preparation of R- beta-hydroxybutyrate or a salt thereof.

37. The pharmaceutical composition of claim 34, wherein the therapeutically effective amount of 2-DG achieves a brain concentration of 2.5-1 OmM.

38. The pharmaceutical composition of claim 34, wherein the therapeutically effective amount of beta-hydroxybutyrate achieves a brain concentration of 15-25mM.

39. The method of any one of claims 27 to 33, wherein the deleterious sequelae are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic stress disorder, or other adverse consequences of the initial traumatic brain injury.

40. The pharmaceutical composition of any one of claims 34 to 38, wherein the deleterious sequelae are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic stress disorder, or other adverse consequences of the initial traumatic brain injury.41 . The method of claim 39, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered acutely to reduce severity and progression of traumatic brain injury and deleterious sequelae that are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic stress disorder, or other adverse consequences of the initial traumatic brain injury.2942. The pharmaceutical composition of claim 40, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones and are administered acutely to reduce severity and progression of traumatic brain injury and deleterious sequelae that are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic during the convulsion or [epileptic] seizure or thereafter.

43. The method of any one of claims 27 to 33, wherein the therapeutically effective amounts of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered chronically to an adult or juvenile animal in need thereof to reduce severity and progression of traumatic brain injury and deleterious sequelae that are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic stress disorder, or other adverse consequences of the initial traumatic brain injury.

44. The pharmaceutical composition of any one of claims 34 to 38, wherein the composition is formulated wherein the therapeutically effective amounts of 2-DG or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones are administered chronically to reduce severity and progression of traumatic brain injury and deleterious sequelae that are structural progression of an initial acute injury, progressive brain tissue loss and structural abnormality, progressive cognitive decline, post-traumatic epilepsy, posttraumatic.

45. The pharmaceutical composition of claims 42 or 44 that is formulated for intravenous or oral administration.

46. The method of claims 41 or 43, wherein the therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones is administered intravenously either simultaneously or sequentially.

47. The pharmaceutical composition of claims 42 or 44, wherein the therapeutically effective amount of 2-DG and beta-hydroxybutyrate or 2-DG in combination with acetone, ketone precursors, metabolic precursors or intermediates, or supplements that increase brain ketones is administered orally either simultaneously or sequentially.30[0001]Statement under Article 19(1)[0002]Applicant submits herewith a replacement sheet containing the claims after amendments. Applicant respectfully requests that the replacement sheet be entered into the application.[0003]Claims 13, 14, 17, 18, 39, 40, 43, and 44 have been amended to refer to the claims in an alternative form to satisfy the second sentence of Rule 6.4(a) and address the objections in the Written Opinion. Claims 19, 21 , 23, and 45 have been amended for typographical errors and formalities. No new matter has been added as a result of these amendments. A marked-up version of the claim amendments is shown in Exhibit A that is attached with this statement.[0004]If a representative of the IB believes it to be helpful to resolve any outstanding issue, the IB is invited to contact the undersigned representative by telephone at +1 3129132101 or by email at ta@mbhb.com.