Methods and compositions for the treatment of alzheimer's disease

Abstract

Disclosed herein are methods and kits for preventing, ameliorating, or treating tauopathies and / or reducing the severity of one or more risk factors, signs, or symptoms associated with Alzheimer's disease.

Description

METHODS AND COMPOSITIONS FOR THE TREATMENT OF ALZHEIMER’S DISEASECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 710,870, filed October 23, 2024, the contents of which are incorporated herein by reference in their entirety.TECHNICAL FIELD

[0002] The present technology relates generally to methods and compositions for preventing, ameliorating, or treating Alzheimer’s disease and / or reducing the severity of one or more risk factors, signs, or symptoms associated with Alzheimer’s disease.BACKGROUND

[0003] The following description is provided to assist the understanding of the reader. None of the information provided or references cited is admitted to be prior art to the compositions and methods disclosed herein.

[0004] Alzheimer's disease is a progressive brain disorder that is characterized by changes in the brain that lead to development amyloid plaques and neurofibrillary tangles.Alzheimer's disease causes brain shrinkage and neuron loss and is the most common cause of dementia, a gradual decline in memory, thinking, behavior and social skills. Approximately 6.5 million people in the United States age 65 and older live with Alzheimer's disease and of the 55 million people worldwide with dementia, 60% to 70% are estimated to have Alzheimer's disease.

[0005] Early symptoms include forgetting recent events or conversations, which eventually worsens to serious memory problems and loss of the ability to perform everyday tasks.Current therapies are extremely limited, and there is no cure for the disease. Accordingly, there is a need for new methods and compositions to treat, prevent, and ameliorate Alzheimer’s disease.SUMMARY

[0006] In one aspect, the present disclosure provides a method for treating or preventing Alzheimer’s disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising D-beta-hydroxybutyrate. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition comprises a pharmaceutically acceptable salt appropriate to the beta-hydroxybutyrate compound.

[0007] In some embodiments, the composition is formulated for intranasal administration. In some embodiments, the method further comprises administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent is selected from the group consisting of anti-amyloid plaque agents, cholinesterase inhibitors, glutamate regulators, orexin receptor antagonists, or atypical antipsychotics. In some embodiments, the antiamyloid plaque agent is selected from the group consisting of aducanumab and lecanemab. In some embodiments, the cholinesterase inhibitor is selected from the group consisting of donepezil, rivastigmine, and galantamine. In some embodiments, the glutamate regulators comprises memantime. In some embodiments, the orexin receptor antagonist comprises suvorexant. In some embodiments, the atypical antipsychotic comprises brexpiprazole. In some embodiments, the additional therapeutic agent is administered simultaneously, sequentially or separately with the composition or is administered as part of the composition. In some embodiments, the method is effective for preventing or reducing one or more signs or symptoms of Alzheimer’s disease. In some embodiments, the one or more signs or symptoms of Alzheimer’s disease is selected from the group consisting of amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof. In some embodiments, the subject is administered the composition at a regular or irregular interval. In some embodiments, the subject is administered the composition daily, weekly, or monthly.

[0008] In one aspect, the present disclosure provides a method for treating or preventing Alzheimer’s disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition comprises a pharmaceutically acceptable salt appropriate to the beta-hydroxybutyrate compound.

[0009] In some embodiments, the composition is formulated for intranasal administration. In some embodiments, the method further comprises administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent is selected from the group consisting of anti-amyloid plaque agents, cholinesterase inhibitors, glutamate regulators, orexin receptor antagonists, or atypical antipsychotics. In some embodiments, the antiamyloid plaque agent is selected from the group consisting of aducanumab and lecanemab. In some embodiments, the cholinesterase inhibitor is selected from the group consisting of donepezil, rivastigmine, and galantamine. In some embodiments, the glutamate regulators comprises memantime. In some embodiments, the orexin receptor antagonist comprises suvorexant. In some embodiments, the atypical antipsychotic comprises brexpiprazole. In some embodiments, the additional therapeutic agent is administered simultaneously, sequentially or separately with the composition or is administered as part of the composition. In some embodiments, the method is effective for preventing or reducing one or more signs or symptoms of Alzheimer’s disease. In some embodiments, the one or more signs or symptoms of Alzheimer’s disease is selected from the group consisting of amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof. In some embodiments, the subject is administered the composition at a regular or irregular interval. In some embodiments, the subject is administered the composition daily, weekly, or monthly.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] FIG. 1 is a graph showing the average body weight over time of wild type control C57B1 / 6 mice treated with saline vehicle (WT Veh), ARTE10 (APP-PS1) mice treated with vehicle (Hm Veh), and ARTE10 (APP-PS1) mice treated with BHB (Hm NaBHB).

[0011] FIG. 2 is a graph showing the average nestlet weight change in a nesting assay for WT Veh, Hm Veh, and Hm NaBHB 180 mg mice, with a greater change indicating more manipulation of the nestlet.

[0012] FIG. 3A is a graph showing the average percentage of pre-pulse inhibition (% PPI) for wild type control C57B1 / 6 mice treated with saline vehicle (WT Veh), ARTE10 (APP-PS1) mice treated with vehicle (Hm Veh), and ARTE10 (APP-PS1) mice treated with BHB (Hm 180), wherein the pre-pulse was at 75 dB and the startle pulse was 120 dB. FIG. 3B is a graph showing the % PPI for WT Veh, Hm Veh, and Hm 180 mice, wherein the pre-pulse was at 80 dB and the startle pulse was 120 dB. FIG. 3C is a graph showing the % PPI for WT Veh, Hm Veh, and Hm 180 mice, wherein the pre-pulse was at 85 dB and the startle pulse was 120 dB.

[0013] FIG. 4A is a graph showing the average running wheel revolutions per hour for wild type control C57B1 / 6 mice treated with saline vehicle (WT Veh), ARTE10 (APP-PS1) mice treated with vehicle (Hm Veh), and ARTE10 (APP-PS1) mice treated with BHB (Hm 180).FIG. 4B is a graph showing the cumulative running wheel revolutions over time for WT Veh, Hm Veh, and Hm 180 mice. FIG. 4C is a graph showing the average running wheel distance covered per hour for WT Veh, Hm Veh, and Hm 180 mice. FIG. 4D is a graph showing the cumulative running wheel distance covered over time for WT Veh, Hm Veh, and Hm 180 mice. FIG. 4E is a graph showing the average running wheel speed per hour for WT Veh, Hm Veh, and Hm 180 mice.DETAILED DESCRIPTION

[0014] It is to be appreciated that certain aspects, modes, embodiments, variations and features of the present technology are described below in various levels of detail in order to provide a substantial understanding of the present technology. The definitions of certainterms as used in this specification are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this present technology belongs.I. Definitions

[0015] The following terms are used herein, the definitions of which are provided for guidance.

[0016] As used herein, the singular forms “a,” “an,” and “the” designate both the singular and the plural, unless expressly stated to designate the singular only.

[0017] The term “about” and the use of ranges in general, whether or not qualified by the term about, means that the number comprehended is not limited to the exact number set forth herein, and is intended to refer to ranges substantially within the quoted range while not departing from the scope of the present technology. As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term.

[0018] As used herein, “administration” of an agent, drug, or composition of the present technology to a subject includes any route of introducing or delivering to a subject a compound to perform its intended function. Administration can be carried out by any suitable route, including orally, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), topically, or by inhalation. In some embodiments, the compositions of the present technology are formulated for enteric administration. In some embodiments, the compositions are formulated for oral, sublingual, buccal, or rectal delivery. In some embodiments, the compositions are formulated for intranasal administration. As used herein, administration includes self-administration and administration by another.

[0019] The terms “beta-hydroxybutyrate, P-hydroxybutyrate”, and “BHB” are used herein interchangeably to refer to the predominant ketone body in the blood or a synthetic BHBcompound. In the body, it is formed mainly in the liver from free fatty acids during fasting or on a high-fat, low-carbohydrate ketogenic diet.

[0020] As used herein, the terms “effective amount,” or “therapeutically effective amount,” and “pharmaceutically effective amount” refer to a quantity sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount which results in the prevention of a disease, condition, and / or symptom(s) thereof. In the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will depend on the type and severity of the disease and on the characteristics of the subject, such as general health, age, sex, body weight, and tolerance to the composition drugs. It will also depend on the degree, severity, and type of disease or condition. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. In some embodiments, multiple doses are administered. Additionally or alternatively, in some embodiments, multiple therapeutic compositions or compounds (e.g., pharmaceutical compositions comprising D-beta-hydroxybutyrate alone or in combination with additional active agents, such as anti-amyloid plaque agents (e.g., aducanumab or lecanemab), cholinesterase inhibitors (e.g., donepezil, rivastigmine, or galantamine) glutamate regulators (e.g., m emantime), orexin receptor antagonists (e.g., suvorexant), or atypical antipsychotics (e.g., brexpiprazole). In the methods described herein, compositions comprising D-beta-hydroxybutyrate of the present technology may be administered to a subject having one or more signs, symptoms, or risk factors of Alzheimer’s disease. Common signs and symptoms of Alzheimer’s disease include amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity (e.g. poor problem solving skills), mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof. For example, a “therapeutically effective amount” of the compositions of the present technology, includes levels at which the presence, frequency, or severity of one or more signs, symptoms, or risk factors of Alzheimer’s disease are, at a minimum, ameliorated. In some embodiments, atherapeutically effective amount reduces or ameliorates the physiological effects of Alzheimer’s disease, and / or the risk factors of Alzheimer’s disease, and / or the likelihood of developing Alzheimer’s disease. In some embodiments, a therapeutically effective amount is achieved by multiple administrations. In some embodiments, a therapeutically effective amount is achieved with a single administration.

[0021] As used herein, “pharmaceutically acceptable carrier and / or diluent” or “pharmaceutically acceptable excipient” includes but is not limited to solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In some embodiments, the pharmaceutically acceptable carrier comprises a polysaccharide, locust bean gum, an anionic polysaccharide, a starch, a protein, sodium ascorbate, glutathione, trehalose, sucrose, or pectin. In some embodiments, the polysaccharide comprises a plant, animal, algal, or microbial polysaccharide. In some embodiments, the polysaccharide comprises guar gum, inulin, amylose, chitosan, chondroitin sulphate, an alginate, or dextran. In some embodiments, the starch comprises rice starch. The use of such media and agents for biologically active substances is well known in the art. Further details of excipients are provided below. Supplementary active ingredients, such as antimicrobials, for example antifungal agents, can also be incorporated into the compositions.

[0022] As used herein, “pharmaceutically acceptable excipient” refers to substances and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal or a human. As used herein, the term includes all inert, non-toxic, liquid or solid fillers, or diluents that do not react with the therapeutic substance of the present technology in an inappropriate negative manner, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, preservatives and the like, for example liquid pharmaceutical carriers e.g., sterile water, saline, sugar solutions, Tris buffer, ethanol, mucoadhesives, permeation enhancers, and / or certain oils.

[0023] As used herein, “prevention,” “prevent,” or “preventing” of a disorder or condition refers to reduction in the occurrence or recurrence of the disorder or condition in treated subjects / samples relative to an untreated controls, or refers delays the onset of one or more symptoms of the disorder or condition relative to the untreated controls.

[0024] As used herein “subject” and “patient” are used interchangeably. In some embodiments, the subject is an animal subject. In some embodiments, the animal subject is a mammal. In some embodiments, the mammalian subject is a human.

[0025] As used herein, the term “simultaneous” administration refers to the administration of at least two agents by the same route and at the same time or at substantially the same time.

[0026] As used herein, the term “separate” administration refers to an administration of at least two agents at the same time or at substantially the same time by different routes.

[0027] As used herein, the term “sequential” administration refers to administration of at least two agents at different times, the administration route being identical or different. More particularly, sequential use refers to the whole administration of one agent before administration of the other agent(s) commences. It is thus possible to administer one of the agents over several minutes, hours, or days before administering another.

[0028] A “synergistic therapeutic effect” refers to a greater-than-additive therapeutic effect which is produced by a combination of at least two therapeutic agents, and which exceeds that which would otherwise result from the individual administration of the agents.

[0029] “Treating,” “treat,” “treated,” or “treatment” of a disease or disorder includes: (i) inhibiting the disease or disorder, z.e., arresting its development; (ii) relieving the disease or disorder, z.e., causing its regression; (iii) slowing progression of the disorder; and / or (iv) inhibiting, relieving, or slowing progression of one or more symptoms of the disease or disorder.

[0030] It is to be appreciated that the various modes of treatment or prevention of medical diseases and conditions as described are intended to mean “substantial,” which includes total but also less than total treatment or prevention, and wherein some biologically or medically relevant result is achieved.II. Alzheimer’s disease

[0031] Alzheimer’s disease is a neurodegenerative disorder characterized by progressive memory loss and the accumulation of deposits (e.g., amyloid plaques, tau deposits, and neurofibrillary tangles). The exact cause of Alzheimer’s is not known, however at present it is thought that a combination of age-related changes in the brain along with genetic, environmental, and lifestyle factors lead to the condition developing. Alzheimer’s disease is further characterized by symptoms including, but not limited to: amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity (e.g. poor problem solving skills), mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof.

[0032] Alzheimer’s is typically diagnosed with a combination of behavioural symptom monitoring, memory tests, cerebrospinal fluid assessment for amyloid and tau levels, and brain scans, such as computed tomography (CT), magnetic resonance imaging (MRI), or positron emission tomography (PET), to support an Alzheimer’s diagnosis or rule out other possible causes for symptoms. Familial medical history and genetic counselling may also be of use. For example, the APOE4 risk allele is associated with higher chances of individuals over the age of 55 years developing Alzheimer’s disease and could indicate the likelihood of developing the disease. Standard medical tests, such as blood and urine tests, or brain scans may be conducted to rule out other possible causes of the symptoms. New and emerging diagnostic methods include structural neuroimaging, molecular imaging, and biofluid biomarker analysis, particularly of cerebrospinal fluid.III. D-Beta-Hydroxybutyrate

[0033] The technology of the present disclosure relates to the use of compositions comprising D-beta-hydroxybutyrate to treat or prevent Alzheimer’s disease. In any of theembodiments described herein D-beta-hydroxybutyrate could be a compound in the form of BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester. In some embodiments, D-beta-hydroxybutyrate is used in methods and compositions for treating or preventing Alzheimer’s disease. In some embodiments, compositions of the present technology comprise D-beta-hydroxybutyrate and a pharmaceutically acceptable carrier. In some embodiments, the D-beta-hydroxybutyrate compositions of the present technology are formulated for intranasal administration. In some embodiments, the composition comprises a pharmaceutically acceptable salt appropriate to the betahydroxybutyrate compound.

[0034] According to some embodiments, the composition when delivered intranasally may elevate brain D-beta hydroxybutyrate levels, e.g., to about 0.1-3 mmol / L, or about 0.1 -1 mmol / L for at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at last 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least 25 hours, at least 26 hours, at least 27 hours, at least 28 hours ,at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, at least 48 hours. According to some embodiments, the composition when delivered intranasally may elevate brain D-beta hydroxybutyrate levels, e.g., to about 2.0 pg / g brain mass to about 3.0 pg / g brain mass for at least 6 hours to at least 48 hours, inclusive, i.e., at least 6 hours, at least 7 hours, at least 8 hours, at last 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least 24 hours, at least 25 hours, at least 26 hours, at least 27 hours, at least 28 hours ,at least 29 hours, at least 30 hours, at least 31 hours, at least 32 hours, at least 33 hours, at least 34 hours, at least 35 hours, at least 36 hours, at least 37 hours, at least 38 hours, at least 39 hours, at least 40 hours, at least 41 hours, at least 42 hours, at least 43 hours, at least 44 hours, at least 45 hours, at least 46 hours, at least 47 hours, at least 48 hours. In some embodiments, thecomposition when delivered intranasally may elevate brain D-beta hydroxybutyrate levels, e.g, to about 2.0 pg / g, 2.1 pg / g, 2.2 pg / g, 2.3 pg / g, 2.4 pg / g, 2.5 pg / g, 2.6 pg / g, 2.7 pg / g, 2.8 pg / g, 2.9 pg / g, or 3.0 pg / g brain mass.

[0035] According to some embodiments, for intranasal delivery, the dose is administered as frequently as tolerated, e.g. every 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 3;8 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours ,61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours etc. to achieve a minimum daily dose. In some embodiments, the minimum daily dose ranges from about 85 mg / day e.g., for a child weighing about 45 lb.) to about 800 mg / day (e.g., for an adult) inclusive, i.e., at least about 85 mg / day; about 95 mg / day; about 105 mg / day, about 115 mg / day; about 125 mg / day; about 135 mg / day; about 145 mg / day; about 155 mg / day; about 165 mg / day; about 175 mg / day; about 185 mg / day; about 195 mg / day; about 205 mg / day, about 215 mg / day; about 225 mg / day; about 235 mg / day; about 245 mg / day; about 255 mg / day; about 265 mg / day; about 275 mg / day; about 285 mg / day; about 295 mg / day; about 305 mg / day, about 315 mg / day; about 325 mg / day; about 335 mg / day; about 345 mg / day; about 355 mg / day; about 365 mg / day; about 375 mg / day; about 385 mg / day; about 395 mg / day; about 405 mg / day, about 415 mg / day; about 425 mg / day; about 435 mg / day; about 445 mg / day; about 455 mg / day; about 465 mg / day; about 475 mg / day; about 485 mg / day; about 495 mg / day; about 505 mg / day, about 515 mg / day; about 525 mg / day; about 535 mg / day; about 545 mg / day; about 555 mg / day; about 565 mg / day; about 575 mg / day; about 585 mg / day; about 595 mg / day; about 605 mg / day, about 615 mg / day; about 625 mg / day; about 635 mg / day; about 645 mg / day; about 655 mg / day; about 665 mg / day; about 675 mg / day; about 685 mg / day; about 695 mg / day; about 705 mg / day, about 715 mg / day; about 725 mg / day; about 735 mg / day; about 745 mg / day; about 755 mg / day; about 765 mg / day; about 775 mg / day; about 785 mg / day; about 795 mg / day; or about 800 mg / day. According to some embodiments, the minimum daily dose for a 60 kghuman (132 lb) is 250 mg / day. According to some embodiments, the minimum daily dose for a 90 kg human (200 lb) is 730 mg / day. According to some embodiments, a dose time curve is determined so that a therapeutically effective dose is delivered intranasally at a given tolerated frequency, e.g., every 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, or 30 days.IV. Therapeutic and Prophylactic Methods

[0036] The following discussion is presented by way of example only, and is not intended to be limiting. One aspect of the present technology includes methods of treating or preventing Alzheimer’s disease in a subject diagnosed as having, suspected as having, or at risk of having Alzheimer’s disease. In therapeutic applications, compositions or medicaments comprising D-beta-hydroxybutyrate are administered to a subject suspected of, or already suffering from Alzheimer’s disease (such as, e.g., subjects exhibiting one or more signs or symptoms of Alzheimer’s disease), in an amount sufficient to cure, or at least partially arrest, the signs or symptoms of Alzheimer’s disease, including its complications and intermediate pathological phenotypes in development of the disease.

[0037] Subjects suffering from Alzheimer’s disease can be identified by any or a combination of diagnostic or prognostic assays known in the art. For example, typical symptoms of Alzheimer’s disease include, but are not limited to: amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof. In some embodiments Alzheimer’s disease is diagnosed using imaging technology to examine the brain and spinal column for abnormalities. In some embodiments, Alzheimer’s disease is diagnosed using genetic testing.

[0038] In one aspect, the present technology provides a method for preventing or delaying the onset of Alzheimer’s disease or symptoms of Alzheimer’s disease in a subject at risk of having Alzheimer’s disease. In some embodiments, subjects with Alzheimer’s disease treated with the compositions of the present technology will show amelioration or elimination of one or more of the following symptoms: amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof.V. Modes of Administration and Effective Dosages

[0039] Compositions of the present technology comprising D-beta-hydroxybutyrate are effective for use in preventing, ameliorating, or treating Alzheimer’s disease and / or reducing the severity of one or more risk factors, signs, or symptoms associated with Alzheimer’s disease. The compositions of the present technology are administered to the subject in effective amounts ( / .< ., amounts that have desired therapeutic effect). The dose and dosage regimen will depend upon the stage of Alzheimer’s disease, the degree of Alzheimer’s disease progression and symptom severity in the subject, the characteristics of the composition used, e.g., its therapeutic index, the subject, and the subject’s history. The effective amount may be determined during pre-clinical trials and clinical trials by methods familiar to physicians and clinicians.

[0040] Administration of the compositions of the present technology can be carried out by any suitable route, such as intranasally, orally, subcutaneously, intraperitoneally, systemically, intravenously, intracerebroventricularly, iontophoretically, transmucosally or intramuscularly. Compositions of the present technology may be formulated for administration by any suitable route. Compositions of the present technology may be formulated for intranasal delivery. Compositions formulated for intranasal administration may be advantageous for subjects with difficulty swallowing.

[0041] The compositions of the present technology can be conveniently provided as sterile, or non-sterile, liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may be buffered to a selected pH. Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) and suitable mixtures thereof.

[0042] Dosage, toxicity and therapeutic efficacy of any therapeutic agent can be determined by standard pharmaceutical procedures in cell cultures or experimental animals. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds may be within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the methods, the therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 ( / .< ., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to determine useful doses in humans accurately.

[0043] An exemplary treatment regimen entails administration once per day. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, or until the subject shows partial or complete amelioration of symptoms of disease. Thereafter, the subject can be administered a prophylactic regime. In some embodiments, compositions of the present technology are administered multiple times per day. In some embodiments, compositions of the present technology are administered to a subject once, twice, or three times per day or more for a certain period of time or until the subject is deemed cured of primary disease, not to be at riskfor recurrence of primary disease, or not to be at risk for the disease. In some embodiments, the compositions of the present technology may be administered to the subject for the remainder of the subject’s life. In some embodiments, administration is paired with an exposure to additional therapeutics (i.e., agents known in the art for the treatment of Alzheimer’s disease, such as anti-amyloid plaque agents (e.g., aducanumab or lecanemab), cholinesterase inhibitors (e.g., donepezil, rivastigmine, or galantamine) glutamate regulators (e.g., memantime), orexin receptor antagonists (e.g., suvorexant), or atypical antipsychotics (e.g., brexpiprazole)), either simultaneously, separately, or sequentially with once, twice, or three times daily dosing of the compositions of the present technology. In some embodiments the compositions of the present technology and the additional therapeutics are administered via the same or different routes of administration. In some embodiments, methods of prophylaxis comprise administration of compositions of the present technology once, twice, or three times daily.

[0044] The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to, the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the therapeutic compositions described herein can include a single treatment or a series of treatments.EXAMPLES

[0045] The following examples are provided by way of illustration only and not by way of limitation. Those of skill in the art will readily recognize a variety of non-critical parameters that could be changed or modified to yield essentially the same or similar results. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims.Example 1: D-beta-hydroxybutyrate Treats and Prevents Alzheimer’s disease

[0046] This example demonstrates that compositions comprising beta-hydroxybutyrate (BHB) are useful in methods for treating and / or preventing Alzheimer’s disease by reducing, ameliorating, or preventing one or more signs and symptoms of Alzheimer’s disease.

[0047] The ARTEIO (APP-PS1) transgenic mouse model is a well-established and translationally relevant tool for Alzheimer’s disease (AD) research. Developed on a C57BL / 6 (B6) background, it co-expresses human APP with the Swedish mutation and PSEN1 with the M146V mutation under the Thyl promoter, resulting in early and robust P-amyloid plaque deposition. By 5 months of age, ARTEIO (APP -PSI) mice display behavioral abnormalities such as hyperactivity, impaired sensorimotor gating, and reduced self-care. At this stage, they also exhibit Alzheimer’ s-relevant biomarkers, including elevated amyloid burden in the hippocampus and cortex, along with emerging changes in neurofilament light chain. In this study, the impact of BHB on both behavioral comorbidities and biomarker profiles in 5-month-old ARTEIO mice was assessed.Materials and Methods

[0048] Study Groups: 5-month-old AD model mice ARTEIO (APP -PSI): B6.CBA-Tg (Thyl-PSEN1*M164V-, APP *S we) 10 Arte and wild type (WT) control C57B1 / 6 Ntac mice were acclimated to the study facility for a minimum of 3 days. Food and water were provided ad libitum in standard housing conditions with bedding, 12-hour light / dark cycles, and enrichment. Mice were house individually. Each treatment group was treated intranasally twice a day for 30 days with either vehicle (saline) or 180 mg / kg (R)-(-)-3-Hydroxybutyric acid sodium salt in saline (NaBHB) as follows: (1) WT C57B1 / 6 mice treated with vehicle, (2) ARTEIO (APP -PSI) mice treated with vehicle, and (3) ARTE 10(APP -PSI) treated with NaBHB (n=12 for all). 17.5 pl per nostril was administered for a total of 35 pl per administration. Weight was measured daily throughout treatment. Nesting tests were performed on day 27 of treatment, the running wheel test was performed on day 28 of treatment, pre-pulse inhibition (PPI) was assayed on day 29 of treatment, and terminal collection of the cortex was performed on the last day of treatmetn (see below for experimental protocol details).

[0049] Nesting Behavior: Nesting is an intrinsic behavior observed in both male and female rodents, even when raised in controlled laboratory environments. Consequently, researchers frequently supply rodents with synthetic and natural materials (e.g., twine, tissue, cotton, paper, hay) to assess their overall well-being and as an auxiliary measure to observe potential cognitive decline. Deviations in nesting behaviors, such as the inability to constructa nest, typically signify alterations in health or welfare. Furthermore, nesting behavior is responsive to various environmental and physiological stressors, as well as genetic mutations associated with pathological conditions, as known in the art. Initial dosing occurred at -8:30 am prior to the start of any tests, and the second dose was performed in the PM after testing was completed at the typical dosing time.

[0050] Mice were individually housed with a nestlet, with an absence of additional bedding, for a duration of 5 hours to evaluate nesting behavior. Undisturbed nesting material remaining after the 5-hour period was weighed to determine the extent of animal engagement with nesting materials.

[0051] Pre-Pulse Inhibition: Pre-pulse inhibition (PPI) of the startle reflex is a well-established neurophysiological measure used in translational research to assess sensorimotor gating. PPI reflects a form of startle plasticity, characterized by a reduced startle response when a weak pre-stimulus (prepulse) precedes a more intense startling stimulus (pulse). This phenomenon has been consistently observed across species, including rodents and humans, as known in the art. The PPI assessment was conducted using the SR-LAB Startle Response System (San Diego Instruments), which consists of an illuminated, ventilated outer chamber designed to minimize external noise and vibration. Inside the chamber, each mouse is placed in a Plexiglas cylinder mounted on a platform equipped with a piezoelectric accelerometer. This sensor detects movement and converts it into digital signals, which are processed and recorded via a connected computer system. Acoustic stimuli are delivered through a calibrated loudspeaker. Prior to testing, mice underwent habituation in the Plexiglas cylinder for 10 minutes on two separate days. On the test day, animals were placed in the cylinder for up to 45 minutes. The stimulation protocol includes background noise (70 dB), startle pulse alone (120 dB), prepulse alone (75, 80, or 85 dB), and combined prepulse + startle trials. Startle responses were recorded and analyzed to evaluate sensorimotor gating and the potential modulatory effects of NaBHB treatment. Non-responders, characterized as those who did not respond to the startle 120 db signal alone, were removed from the analysis (n = 4 in WT mice; n = 0 in vehicle-only ARTE10 (APP-PS1) mice; n = 1 in BHB-treated ARTE10 (APP-PS1) mice). Initial dosing occurred at -8:30 am prior to the start of the test, and the second dose was performed in the PM and only after testing was completed.

[0052] Running wheel: Wheel running is believed to reflect multiple underlying behavioral processes beyond general spontaneous activity, offering valuable insights into circadian rhythms, fatigue or depression-like states, and anxiety-related behaviors. Given that ARTE10 mice exhibit heightened anxiety, assessing hyperactivity in a familiar environment using running wheels may provide a more accurate measure than traditional tests such as the Locomotor Activity (LMA) assay, which can be more anxiety-inducing and potentially confound results. Additionally, running wheel activity can reveal disruptions in circadian patterns, which are commonly associated with AD. In this study, wheel running was used to evaluate hyperactivity characteristic of the ARTE10 phenotype and to determine whether treatment with NaBHB mitigates these behavioral alterations. Mice had access to running wheels for a 20-hour period, during which activity was continuously recorded and analyzed over time, including wheel revolutions per hour, cumulative revolutions, distance per hour, cumulative distance traveled, and speed per hour. Initial dosing occurred at -8:30 am, animals were then placed in the running wheel cages by 12:00 and momentarily removed from the testing cages for the second dose in the PM. The running wheel for a single mouse in the vehicle-only ARTE10 (APP-PS1) group malfunctioned, and that mouse was therefore excluded from the analysis resulting in an n = 11 for that treatment group in this experiment.

[0053] Terminal Collection and Assessment of Neurofilament light chain and amyloid beta:Following the final day of behavioral testing animals underwent terminal collections.Animals received their first daily dose on collection day and were humanely euthanized by inhaled anesthesia overdose (isoflurane 4-5%) followed by decapitation for brain collection. Brains were dissected and cortexes were isolated before being flash frozen in liquid Nitrogen. Neurofilament light chain and amyloid beta abundance in the cortex will be assessed using standard methodologies.Results

[0054] The results of the experiments described above demonstrate a clear improvement to the behavioral and physical symptoms of Alzheimer’s disease in BHB-treated ARTE10 (APP-PS1) mice disease model mice.

[0055] Weight loss is a well-known symptom of Alzheimer’s disease that occurs throughout the course of disease progression. As shown in FIG. 1, BHB-treated ARTEIO (APP-PS1) mice had consistently higher body weights than ARTEIO (APP-PS1) mice that received vehicle alone.

[0056] Nesting behavior was also improved in BHB-treated ARTEIO (APP-PS1) mice as compared to vehicle-only ARTEIO (APP-PS1) mice. Treated mice demonstrated a substantial increase in nestlet weight change at the end of the 5-hour test period, reflecting increased interaction with the nesting material (FIG. 2).

[0057] Pre-pulse inhibition results also demonstrated consistent improvement in BHB-treated ARTEIO (APP-PS1) mice (“HM 180” in FIG. 3A-FIG. 3C) compared to control vehicle-only ARTEIO (APP-PS1) mice (“HM Veh” in FIG. 3A-FIG. 3C). At each pre-pulse volume tested of 75dB (FIG. 3A), 80dB (FIG. 3B), and 85dB (FIG. 3C), BHB-treated ARTEIO (APP-PS1) mice had an improved PPI compared to vehicle-only control mice, indicating an improved startle response.

[0058] Running wheel test results further corroborated the results of the body weight, nesting, and PPI assessments in demonstrating a therapeutic effect from BHB treatment in AD model mice. As shown in FIG. 4A- FIG. 4E, BHB-treated ARTEIO (APP-PS1) mice (“HM 180”) had substantially reduced hourly (FIG. 4A) and cumulative (FIG. 4B) running wheel rotations, hourly (FIG. 4C) and cumulative (FIG. 4D) distance traveled, and hourly speed (FIG. 4E), compared to vehicle-only ARTEIO (APP-PS1) mice (“HM Veh”). At multiple time points during the course of the 20-hour assay period, BHB-treated ARTEIO (APP-PS1) mice exhibited statistically significant decreases in hourly and cumulative wheel rotations, hourly and cumulative distance traveled, and hourly speed (data not shown) compared to vehicle-only ARTEIO (APP-PS1) mice. BHB-treated ARTEIO (APP-PS1) mice behaviors were indistinguishable from wild type vehicle-only mice (“WT Veh”), a compelling result considering that treatment was limited to a 30-day window.

[0059] It is anticipated that the results of the beta amyloid and neuro filament light chain analysis will demonstrate that BHB treatment of ARTEIO (APP-PS1) mice reduces betaamyloid and / or neuro filament light chain cortex abundance compared to vehicle-only ARTEIO (APP-PS1) mice.

[0060] These results demonstrate a clear improvement in physical and behavioral symptoms in BHB-treated ARTEIO (APP-PS1) mice as compared to vehicle-only control ARTEIO (APP-PS1) mice, with BHB treatment reducing weight loss, improving nesting behavior, decreasing the startle response, and reducing hyperactivity. Accordingly, these results show that BHB is therapeutically effective for treating Alzheimer’s disease in a subject in need thereof by reducing one or more signs or symptoms of the disease.Example 2: D-beta-hydroxybutyrate Compounds Treat and Prevent Alzheimer’s disease

[0061] This example will demonstrate that compositions comprising the D-beta-hydroxybutyrate compounds, BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester, are useful in methods for treating and / or preventing Alzheimer’s disease by reducing, ameliorating, or preventing one or more signs and symptoms of Alzheimer’s disease.

[0062] Briefly, the experiments described in Example 1 will be repeated using one or more of BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester in place of D-beta-hydroxybutyrate. Treatment groups will include wild type C57B1 / 6 control mice treated with vehicle alone, ARTEIO (APP-PS1) mice treated with vehicle alone, and ARTEIO (APP-PS1) mice treated with BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester, with all treatments administered intranasally. Treatment will begin at 5-months of age, when ARTEIO (APP-PS1) mice typically begin exhibiting symptoms of AD. Mice will be assessed for one or more of body weight, nesting behavior, pre-pulse inhibition or an equivalent assay for measuring the startle response, running wheel usage or an equivalent assay for measuring hyperactivity, and cortex levels of amyloid beta and neurofilament light chain.

[0063] These results are predicted to show that treatment of AD model ARTEIO (APP-PS1) mice with BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester will result in reduced weight loss, improved nesting behavior, decreased startle response, reduced hyperactivity, and reduced cortex levels of amyloid beta and neurofilamentlight chain as compared to vehicle-only controls. Accordingly, these results will demonstrate that compositions comprising BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, or BHB dimer - L-tyrosine ester are useful in methods for the treatment or prevention of Alzheimer’s disease.EQUIVALENTS

[0064] The present technology is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present technology is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0065] Each and every publication and patent mentioned in the above specification is herein incorporated by reference in its entirety for all purposes. Various modifications and variations of the described methods and system of the present technology will be apparent to those skilled in the art without departing from the scope and spirit of the present technology. Although the present technology has been described in connection with specific embodiments, the present technology as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the present technology which are obvious to those skilled in the art and in fields related thereto are intended to be within the scope of the following claims.

Claims

CLAIMSWhat is claimed is:

1. A method for treating or preventing Alzheimer’s disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising D-beta-hydroxybutyrate or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier.

3. The method of claims 1 or 2, wherein the composition is formulated for intranasal administration.

4. The method of any one of claims 1-3, wherein the method further comprises administering an additional therapeutic agent.

5. The method of claim 4, wherein the additional therapeutic agent is selected from the group consisting of anti-amyloid plaque agents, cholinesterase inhibitors, glutamate regulators, orexin receptor antagonists, or atypical antipsychotics.

6. The method of claim 5, wherein the anti-amyloid plaque agent is selected from the group consisting of aducanumab and lecanemab.

7. The method of claim 5, wherein the cholinesterase inhibitor is selected from the group consisting of donepezil, rivastigmine, and galantamine.

8. The method of claim 5, wherein the glutamate regulators comprises memantime.

9. The method of claim 5, wherein the orexin receptor antagonist comprises suvorexant.

10. The method of claim 5, wherein the atypical antipsychotic comprises brexpiprazole.

11. The method of any one of claims 5-10, wherein the additional therapeutic agent is administered simultaneously, sequentially or separately with the composition or is administered as part of the composition.

12. The method of any one of claims 1-11, wherein the method is effective for preventing or reducing one or more signs or symptoms of Alzheimer’s disease.

13. The method of claim 12, wherein the one or more signs or symptoms of Alzheimer’s disease is selected from the group consisting of amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof.

14. The method of any one of claims 1-13, wherein the subject is administered the composition at a regular or irregular interval.

15. The method of claim 14, wherein the subject is administered the composition daily, weekly, or monthly.

16. A method for treating or preventing Alzheimer’s disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a compound selected from the group consisting of BHB dimer-adenine hemiaminal, BHB dimer-adenine amide, BHB dimer - L-tyrosine ester, and pharmaceutically acceptable salts thereof appropriate to the compound.

17. The method of claim 16, wherein the composition further comprises a pharmaceutically acceptable carrier.

18. The method of claims 16 or 17, wherein the composition is formulated for intranasal administration.

19. The method of any one of claims 16-18, wherein the method further comprises administering an additional therapeutic agent.

20. The method of claim 19, wherein the additional therapeutic agent is selected from the group consisting of anti-amyloid plaque agents, cholinesterase inhibitors, glutamate regulators, orexin receptor antagonists, or atypical antipsychotics.

21. The method of claim 20, wherein the anti-amyloid plaque agent is selected from the group consisting of aducanumab and lecanemab.

22. The method of claim 20, wherein the cholinesterase inhibitor is selected from the group consisting of donepezil, rivastigmine, and galantamine.

23. The method of claim 20, wherein the glutamate regulators comprises memantime.

24. The method of claim 20, wherein the orexin receptor antagonist comprises suvorexant.

25. The method of claim 20, wherein the atypical antipsychotic comprises brexpiprazole.

26. The method of any one of claims 20-25, wherein the additional therapeutic agent is administered simultaneously, sequentially or separately with the composition or is administered as part of the composition.

27. The method of any one of claims 16-26, wherein the method is effective for preventing or reducing one or more signs or symptoms of Alzheimer’s disease.

28. The method of claim 27, wherein the one or more signs or symptoms of Alzheimer’s disease is selected from the group consisting of amyloid plaques, neurofibrillary tangles, tau deposits, progressive memory loss, poor judgement, diminished mental capacity, mood or personality changes, increased anxiety, increased aggression, antisocial behavior, inability to learn new things, difficulty using language, difficulty performing math, diminished attention span, changes in sleeping patters, difficulty performing complex tasks, hallucinations, delusions, paranoia, impulsive behavior, emotional outbursts, repetitive statements or movements, weight loss, diminished appetite, seizures, loss of bladder control, inability to communicate, or any combination thereof.

29. The method of any one of claims 16-28, wherein the subject is administered the composition at a regular or irregular interval.

30. The method of claim 29, wherein the subject is administered the composition daily, weekly, or monthly.

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