How to reverse aging-related brain decline

Abstract

Methods for treating brain decline during aging are provided, comprising administration of a glucagon-like peptide-1 receptor (GLP-1) agonist (GLP-1RA) to treat brain aging-related changes. The GLP-1R agonist is exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, taspoglutide, PF-06882961, OWL-833, TTP-273 or any other molecule that activates GLP-1R.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 126,122, filed December 16, 2020, and is incorporated by reference in its entirety, including any tables, figures, or drawings. [Background technology]

[0002] Aging has been considered an irreversible process. Nearly all cellular processes are involved in and affected by aging, ranging from metabolism, stress response, immune response, cellular senescence, to gene expression and genome stability (Non-Patent Document 1). These complex molecular alterations likely result in alterations of cellular state and composition in many body organs, manifesting as age-related functional decline. In the brain, hallmarks of aging include (1) mitochondrial dysfunction, (2) dysregulation of energy metabolism, (3) intracellular accumulation of oxidatively damaged proteins, nucleic acids, and lipids, (4) impaired cellular "waste disposal" mechanisms (i.e., autophagy, lysosomal, and proteasome functionality), (5) impaired adaptive stress response signaling, (6) impaired DNA repair, (7) abnormalities in neural network activity, (8) dysregulation of neuronal calcium signaling and coping, (9) stem cell depletion, and (10) inflammation (Non-Patent Document 15). Given the complexity of the biological changes involved and the lack of a set of easily targetable driving pathways, anti-aging drug therapy is considered to be very challenging. Molecular and cellular alterations in the aging brain lead to its decline in function and predispose it to stroke and neurodegeneration (e.g., Alzheimer's and Parkinson's disease). As human life spans increase, the population suffering from aging-associated brain conditions is growing dramatically. Slowing down or even reversing these alterations in the aging brain may provide a strategy for the primary prevention or even treatment of these conditions.

[0003] Glucagon-like peptide-1 (GLP-1) is a peptide hormone produced peripherally by intestinal L-cells and centrally in the brain by preproglucagon neurons of the nucleus of the solitary tract to enhance glucose-dependent insulin release (Non-Patent Document 11). In the past decade, a number of pharmacokinetically optimized GLP-1 receptor (GLP-1R) agonists (GLP-1RAs) have been approved for the clinical treatment of diabetes mellitus. Apart from the treatment of diabetes mellitus, evidence in cellular and animal models of neurodegeneration supports the neurotrophic and neuroprotective role of GLP-1R stimulation, as well as in increasing neurogenesis (Non-Patent Document 17, Non-Patent Document 9, Non-Patent Document 7, Patent Document 1, Non-Patent Document 12, Non-Patent Document 7), indicating that GLP-1R is involved in learning and neuroprotection. Intracerebroventricular (icv) administration of GLP-1 and [Ser(2)]exendin(1-9) to 8-week-old rats enhances associative and spatial learning via GLP-1R. Peripheral administration of [Ser(2)]exendin(1-9) is also active. Isacson et al. (Non-Patent Document 12) showed that intraperitoneal (ip) injection of exendin-4 improved hippocampal-associated baseline memory performance and reduced immobility in the forced swimming test. Of note, recent clinical studies have provided compelling evidence that GLP-1RAs exhibit neuroprotective effects beyond those provided by glycemic control and reduce the incidence of cognitive decline and Parkinson's disease (PD) in diabetic patients (Non-Patent Document 6, Non-Patent Document 4). Furthermore, GLP-1RAs can slow the progression of established Alzheimer's disease (AD) and PD in non-diabetic patients (Non-Patent Document 8, Non-Patent Document 2). Mechanistically, apart from reducing neuroinflammation in animal models of neurodegeneration (Non-Patent Document 5, Non-Patent Document 19), Zhao et al. demonstrated that treatment with exenatide (GLP-1RA) partially reversed age-related transcriptomic changes in brain endothelial cells (ECs) and reduced non-specific blood-brain barrier (BBB) ​​leakage (Non-Patent Document 20).

[0004] Aging is the process of growing older. The term specifically refers to mammals. In a broader sense, aging can refer to a single cell or organ in an organism. In humans, aging represents the accumulation of human changes over time. Aging is a major risk factor for cancer, cardiovascular disease, and neurodegenerative diseases, which are prevalent diseases in developed countries (Non-Patent Document 16). The cause of aging is unknown. One current theory is the damage hypothesis, which states that accumulated damage leads to a gradual loss of physiological integrity, leading to increased vulnerability to functional impairment and death (Non-Patent Document 14).

[0005] Aging involves almost all cell types and cellular processes. Moreover, the characteristics of the aging brain are different from those of neurodegenerative diseases with specific symptoms, such as AD, PD and Huntington's disease. Given the complexity of the biological changes involved and the lack of a set of easily targetable driving pathways, anti-aging drug therapy is considered to be very difficult, if possible. Thus, there is a need to slow down or even reverse the transcriptomic and functional alterations in the aging brain. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] U.S. Patent Publication No. 6,969,702 B2 [Non-patent literature]

[0007] [Non-Patent Document 1] Almanzar et al., A single-cell transcriptomic atlas characterizes aging tissues in the mouse. Nature 583, 590-595 (2020) [Non-Patent Document 2] Athauda et al., Exenatide once weekly placebo versus in Parkinson's disease: a randomized, double-blind, placebo-controlled trial. Lancet (London, England) 390, 1664-1675 (2017) [Non-Patent Document 3] Boisvert, MM, GA Erikson, MN Shokhirev, NJ Allen, The Aging Astrocyte Transcriptome from Multiple Regions of the Mouse Brain. Cell Reports 22, 269-285 (2018) [Non-Patent Document 4] Brauer et al., Diabetes medications and risk of Parkinson's disease: a cohort study of patients with diabetes. Brain (2020) https: / doi.org / 10.1093 / brain / awaa262 [Non-Patent Document 5] Cai et al., Lixisenatide reduces amyloid plaques, neurofibrillary tangles and neuroinflammation in an APP / PS1 / tau mouse model of Alzheimer's disease. Biochem Bioph Res Co 495, 1034-1040 (2018) [Non-Patent Document 6] Cukierman-Yaffe et al., Effect of dulaglutide on cognitive impairment in type 2 diabetes: an exploratory analysis of the REWIND trial. Lancet Neurology 19, 582-590 (2020)

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

[0008] Described herein is a method of treating a subject for aging-associated brain symptoms.An embodiment of the method includes administering a GLP-1R agonist to a subject in need thereof, for example, an individual at risk of developing or suffering from aging-associated brain symptoms.Age-associated brain symptoms include, for example, cognitive impairment and structural, functional or molecular changes associated with aging of the brain.

[0009] In certain embodiments, the methods include treatment of a subject who may be at risk of developing or suffering from a brain condition caused by natural aging.

[0010] In certain embodiments, a subject may be at risk of developing or suffering from an aging-associated brain condition that has transcriptomic and functional changes across multiple cell types in the brain, including neurons, such as mature neurons (mNeur) and immature neurons (imNeur); glial cells, such as astrocytes (AC), oligodendrocyte precursor cells (OPC), microglia (MG), and oligodendrocytes (OLG); mural cells, such as pericytes (PC) and smooth muscle cells (SMC) cells; choroid plexus cells (CPC); hemoglobin-expressing vascular cells (Hb_EC); and monocytes (MNC).

[0011] In certain embodiments, the subject may be at risk of developing or suffering from an age-associated brain condition treatable with a GLP-1R agonist (GLP-1RA), such as, for example, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, taspoglutide, PF-06882961, OWL-833 and / or TTP-273.

[0012] In certain embodiments, GLP-1RA treatment can reverse and / or inhibit transcriptomic and functional changes exhibited in multiple cell types in the brain, including neurons, such as mature neurons (mNeur) and immature neurons (imNeur); glial cells, such as astrocytes (AC), oligodendrocyte precursor cells (OPC), microglia (MG) and oligodendrocytes (OLG); mural cells, such as pericytes (PC) and smooth muscle cells (SMC) cells; choroid plexus cells (CPC); hemoglobin-expressing vascular cells (Hb_EC); and monocytes (MNC). [Brief description of the drawings]

[0013] [Figure 1A] UMAP visualization of the major cell type clusters identified and analyzed in mouse brain. AC: astrocytes; OPC: oligodendrocyte precursor cells; MG: microglia; MAC: perivascular macrophages; OLG: oligodendrocytes; SMC: smooth muscle cells; PC: pericytes; EC: endothelial cells. Numbers in brackets: cell number of each cell type. [Figure 1B] UMAP visualization of single cell transcriptomes from young adult mouse brains. Numbers in brackets: group cell number (n=3 per group). [Figure 1C] UMAP visualization of single cell transcriptomes from aged adult mouse brains. Numbers in brackets: group cell number (n=3 per group). [Figure 1D] Figure 1 shows UMAP visualization of single cell transcriptomes from GLP-1RA (exenatide)-treated old mouse brains. Numbers in brackets: cell number of the group (n=3 per group). [Figure 2A] Age-related expression changes (x-axis) plotted against expression changes (y-axis) following GLP-1RA treatment in glial (AC: astrocytes; OPC: oligodendrocyte precursor cells; MG: microglia; and OLG: oligodendrocytes), vascular (EC: endothelial cells; PC: pericytes; SMC: smooth muscle cells) cell types, and MAC (perivascular macrophages). Each point represents one differentially expressed gene (DEG). Grey line: line of best fit from linear regression. [Figure 2B] The percentage of inverted DEGs shown in Figure 2A in different cell types and the slope of the best fit line by linear regression are shown. [Diagram 3] Age-related expression changes (x-axis) plotted against expression changes (y-axis) following GLP-1RA treatment in neurons (mNeur: mature neurons; imNeur: immature neurons; MG: microglia; and OLG: oligodendrocytes), monocytes (MNCs), choroid plexus cells (CPCs) and hemoglobin-expressing vascular cells. Each point represents one differentially expressed gene (DEG). Grey line: line of best fit from linear regression. [Figure 4] Figure 1 shows functional pathways with significant enrichment among the most prominent age-related expression changes reversed by GLP-1RA treatment in different brain cell types. [Diagram 5] FIG. 1 shows changes in expression of selected functionally important genes in AC, MG and SMC during aging and after GLP-1RA treatment. [Figure 6] Figure 1 shows the improvement of spatial memory in aged mice treated with GLP-1RA in the Y-maze test. C57BL / 6 mice aged 15-17 months were treated daily with saline vehicle or 5 nmol / kg body weight exenatide for 2-4 months. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] definition Ranges provided herein are understood to be abbreviated to all values ​​within the range. For example, the range 1-20 is understood to include any number, combination of numbers or subranges from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, as well as all intervening decimal values ​​between the aforementioned integers, such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to subranges, "nested subranges" extending from either end of the range are specifically contemplated. For example, nested subranges of the exemplary range 1-50 can include 1-10, 1-20, 1-30, and 1-40 in one direction, and 50-40, 50-30, 50-20, and 50-10 in the other direction.

[0015] As used herein, "decrease" means a negative alteration and "increase" means a positive alteration, where negative or positive alteration means a positive change of at least 0.001%, 0.01%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[0016] The transitional phrase "comprising," which is synonymous with the transitional phrases "including" or "containing," is inclusive or open-ended and does not exclude additional unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes elements, steps, or ingredients specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to certain materials or steps "and that do not materially affect the basic and novel characteristics" of the claimed invention. Use of the term "comprising" contemplates other embodiments that "consist" or "consist essentially of" the recited components.

[0017] Unless otherwise specified or clear from context, the term "or" as used herein is understood to be inclusive. Unless otherwise specified or clear from context, the terms "a" and "an" as used herein are understood to be singular or plural.

[0018] Unless otherwise specified or clear from the context, the term "about" as used herein is understood to be within normal tolerances in the art, for example, within 2 standard deviations of the mean. About can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05% or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values ​​provided herein are modified by the term about.

[0019] As used herein, the term "pharmaceutical acceptable" means compatible with other ingredients of a pharmaceutical composition and not deleterious to the recipient thereof.

[0020] As used herein, the terms "therapeutically effective amount", "therapeutically effective dose", "effective amount" and "effective dose" are used to refer to an amount or dose of a compound or composition that, when administered to a subject, can treat or ameliorate a symptom, disease or illness of the subject, or provide enhanced health or function to an organ, tissue or body system. In other words, when administered to a subject, the amount is "therapeutically effective". The actual amount will vary depending on many factors, including, but not limited to, the particular symptom, disease or illness being treated or ameliorated; the severity of the symptom; the particular organ, tissue or body system in which an enhancement in health or function is desired; the patient's weight, height, age and health, and the route of administration.

[0021] As used herein, "treatment," "treating," "palliating," and "ameliorating" (and grammatical variations of these terms) are used interchangeably. These terms refer to an approach for obtaining a beneficial or desired result, including, but not limited to, a therapeutic benefit. A therapeutic benefit is achieved by eradicating or ameliorating one or more of the physiological symptoms associated with an underlying disease such that an improvement is observed in the subject, even though the subject may still be afflicted with the underlying disease. Treatment includes delaying the appearance of a disease or condition, delaying the onset of symptoms of a disease or condition, slowing the progression of a disease or condition, slowing, arresting or reversing the progression of symptoms or any combination thereof, or any combination thereof. The term refers to eradicating, reducing, ameliorating or reversing to some extent the signs or symptoms of the underlying disease, including, but not required, curing the condition, disease or illness. Treatment can be a cure, amelioration or partial amelioration of a disease. "Treatment" can also include, for example, the functioning of a particular system in the body to an enhanced state of health or homeostasis.

[0022] As used herein, the term "elderly" refers to a subject in the age group beyond middle age. Elderly refers to a subject in a specific age group, for example, 50 years or older, 60 years or older, 65 years or older, 70 years or older, or 75 years or older. The elderly subject is preferably a mammal, more preferably a human, and even more preferably an adult human, at least 50, 55, 60, 65, or 70 years old. More preferably, the patient is a geriatric or elderly geriatric (adult) human patient. The elderly patient may be male or female.

[0023] As used herein, the phrase "executive function" refers to a set of cognitive abilities that control or regulate other abilities or behaviors. Executive functions are high-level abilities that affect more basic abilities such as attention, memory, and motor skills. These executive functions are necessary for goal-directed behavior and include the ability to initiate or stop actions, to monitor and modify behavior as needed, and to plan future behavior when faced with novel tasks or situations. Executive functions allow subjects to predict outcomes and adapt to changing situations. The ability to form concepts and the ability to think abstractly are often considered components of executive functions.

[0024] Brain symptoms associated with aging Along with other organs, the function of the brain gradually declines with age, and the brains of the aging population are at risk of developing or suffering from age-related conditions that manifest as declines in learning and memory, attention, speed of decision-making, sensory perception, and motor coordination.

[0025] In certain embodiments, a subject who would benefit from a treatment as disclosed herein is at least 50, 60, 70, 80, 90 years of age, typically not exceeding 100 years of age, about 50-100 years of age, about 50 years of age, about 55 years of age, about 60 years of age, about 65 years of age, about 70 years of age, about 75 years of age, about 80 years of age, about 85 years of age, about 90 years of age, or about 100 years of age, and is at risk of developing or suffering from a symptom associated with aging, such as, for example, cognitive impairment and / or measurable structural, functional, or molecular changes in the brain.

[0026] In certain embodiments, cognitive impairment can include various brain symptoms, such as attention and concentration; learning tasks and concepts; memory; information processing; visuospatial function; speech; language comprehension; speech fluency; problem solving; decision making; and executive function. Cognitive impairment can be assessed by a number of tests, such as memory, recall, visuospatial awareness, speech fluency, expression, executive function, gait, and dual tasks. Tests used to detect such differences include, for example, the Mini-Mental State Examination (MMSE), the Montreal Cognitive Assessment (MoCA) and its variants, the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), and the Clinical Dementia (CDR) scale. Scores that meet any of the criteria: 26 or less on the MMSE, 25 or less on the MoCA, 12 or more on the ADAS-Cog, or 0.5 or more on the CDR are considered cognitively impaired.

[0027] In certain embodiments, the structural changes in the brain can be a decrease in brain volume, i.e., 5%, 10%, 15%, 20% or more, changes in the structure of gray matter, the structure of white matter, the structure of the ventricular system, the structure and integrity of the neurovascular system. Structural changes in the brain can be measured, for example, by imaging methods such as magnetic resonance imaging (MRI), computed tomography (CT) or ultrasound (US). Age-related declines in these measurements are defined as alterations in the values ​​and / or patterns obtained by the respective imaging or recording methods that reflect changes in the volume of the structure, falling into the extremes of the distribution for the whole population or population of the same age (e.g., top or bottom 5%, 10%, 20%, 25%, 30%), or showing changes in repeated measurements from the same subject over time (e.g., 5%, 10%, 15%, 20%, 25% or more).

[0028] In certain embodiments, functional changes in the brain can refer to measurable changes in the brain that reflect an altered state of neuronal activity, metabolism, or neurovascular function. Functional changes in the brain can be measured, for example, by functional magnetic resonance imaging (fMRI), magnetic resonance imaging (MRSI), hyperpolarized carbon-13 ( 13C) It can be measured by imaging methods such as magnetic resonance imaging (MRSI), ultrasonography (US), positron emission tomography (PET) or single photon emission computed tomography (SPECT). Functional changes in the brain can also be recorded by electrophysiological methods, such as electroencephalography (EEG), intracranial electrode recording and deep brain electrode recording, or magnetic field-based recording methods, such as magnetoencephalography (MEG). Age-related decline in these measurements is defined as an alteration in the values ​​and / or patterns obtained by the respective imaging or recording methods, reflecting neuronal activity, metabolism or neurovascular function, falling into the extremes of the distribution for the whole population or a population of the same age (e.g., top or bottom 5%, 10%, 20%, 25%, 30%), or exhibiting changes in repeated measurements from the same subject over time (e.g., 5%, 10%, 15%, 20%, 25% or more).

[0029] In certain embodiments, brain molecular changes can refer to molecular level measurable modifications in brain tissue or blood, such as gene expression; DNA transcription; RNA translation; DNA, RNA or protein location in brain cells or tissues, and / or changes in the secretion or release of proteins, DNA, RNA, mitochondria, cellular components or mitochondrial components into blood. The body fluid can be cerebrospinal fluid (CSF), blood or plasma. Age-related changes in these measurements are defined as changes in the values ​​and / or patterns obtained by the respective methods that reflect brain molecular changes, falling into the extremes of the distribution (e.g., top or bottom 5%, 10%, 20%, 25%, 30%) for the whole population or population of the same age, or showing changes in repeated measurements from the same subject over time (e.g., 5%, 10%, 15%, 20%, 25% or more).

[0030] GLP-1R agonists In certain embodiments, GLP-1R agonists can be used in the subject methods.GLP-1R agonists are agonists of the GLP-1 receptor.GLP-1, the natural agonist of GLP-1R, has a short duration of action.Several pharmacologically optimized GLP-1R agonists have been approved or developed for the clinical treatment of diabetes mellitus or neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease.

[0031] The activity of GLP-1R agonists can be determined by assays of well-studied downstream signaling and regulatory pathways associated with GLP-1R activation, such as increased cAMP production, induction of phosphorylation of ERK1 / 2, enhanced intracellular mobilization of calcium, and recruitment of beta-arrestin-1 and beta-arrestin-2.

[0032] Some non-limiting examples of GLP-1R agonists include exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, taspoglutide, PF-06882961, OWL-833, and TTP-273.

[0033] Dosage and Administration In certain embodiments, the methods described herein provide a method for treating brain symptoms associated with aging. In one embodiment, the subject can be a mammal. In other embodiments, the subject can be a human, although the present invention is effective for all mammals. The method can be to administer to the subject an effective amount of a pharmaceutical composition comprising a GLP-1R agonist that reverses transcriptomic and functional changes in the aging brain. The composition can further comprise one or more pharmaceutically acceptable carriers and / or excipients, and can further comprise formulating the composition into, for example, solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols. Pharmaceutically acceptable carriers are well known in the art, and are, for example, aqueous solutions, water or buffered saline or other solvents or vehicles, such as polyethylene glycol, Tween-20 or olive oil or injectable organic esters. The dose range of the agonist can depend on the potency. In certain embodiments, large doses of agonist can produce the desired effect, such as reversal of transcriptomic and functional changes in any major cell type in the aging brain, and reversal of structural and / or functional changes in the aging brain. The dose should not be so large as to cause adverse side effects. In general, the dose will vary depending on the agent used. Furthermore, the age, symptoms and sex of the subject can be determined by those skilled in the art and used to determine the dose. The dose can also be adjusted by the individual physician in the case of complications.

[0034] In one embodiment, the composition is formulated as an orally ingestible product, such as a food product, capsule, pill, or drinkable liquid. An orally deliverable health-promoting compound is any bioactive substance that is delivered via initial absorption into the gastrointestinal or oral mucosa. The composition can also be formulated as a solution that can be administered by injection, including, for example, intravenous, intraperitoneal, intramuscular, intrathecal, or subcutaneous. In another embodiment, the composition is formulated to be administered through a patch or directly through the skin for local or systemic effects. The composition can also be administered sublingually, bucally, rectally, or vaginally. Additionally, the composition can be sprayed, atomized, inhaled through the mouth or nose, or administered to the eye or ear for absorption through the nasal mucosa.

[0035] An orally ingestible product according to the present invention is a preparation or composition suitable for ingestion, nutrition, oral hygiene or palatability, which is placed in the oral cavity of a human or animal and remains there for a period of time before being swallowed (e.g., foods for ingestion or pills) or removed again from the oral cavity (e.g., chewing gum or oral hygiene or medicated mouthwash products). Although an orally deliverable pharmaceutical product can be formulated into an orally ingestible product, and an orally ingestible product can include an orally deliverable pharmaceutical product, the two terms are not meant to be used interchangeably herein.

[0036] Consumable products include any substance or product that is intended to be ingested by humans or animals in a processed, semi-processed or raw state, including those that are added to consumable products (in particular foods and medicines) during their manufacture, treatment or handling and that are introduced into the oral cavity of humans or animals.

[0037] Orally ingestible products can also include materials that are intended to be swallowed and digested by humans or animals in an unmodified, prepared or processed state. Orally ingestible products also include casings, coatings or other encapsulations that are intended or anticipated to be swallowed along with the product.

[0038] In one embodiment, the orally consumable product is a capsule, pill, syrup, emulsion or suspension containing the desired orally deliverable substance, hi one embodiment, the orally consumable product can include the orally deliverable substance in a powder form, which can be mixed with water or other liquid to produce a drinkable orally consumable product.

[0039] Carriers and / or excipients according to the present invention may be any solvent, diluent, buffer (such as neutral buffered saline, phosphate buffer, or optionally Tris-HCl, acetate or phosphate buffer), oil-in-water or water-in-oil emulsions, e.g., aqueous compositions with or without organic co-solvents suitable for IV use, solubilizers (e.g., polysorbate 65, polysorbate 80), colloids, dispersion media, vehicles, fillers, chelating agents (e.g., EDTA or glutathione), amino acids (e.g., glycine), proteins, disintegrants, binders, lubricants, wetting agents, emulsifiers, sweeteners, colorants, flavorings, fragrances, thickening agents (e.g., carbomer, gelatin or sodium alginate), coatings, preservatives (e.g., thimerosal, benzyl alcohol, polyquaternium), antioxidants (e.g., ascorbic acid, sodium metabisulfite), tonicity controlling agents, The composition may include, for example, glyceryl stearate ...

[0040] In one embodiment, the composition can be made into an aerosol formulation, e.g., so that it can be nebulized or inhaled. Pharmaceutical formulations suitable for administration in the form of an aerosol or spray are, for example, solutions, suspensions, or emulsions. Formulations for oral or nasal aerosol or inhalation administration can also be formulated with exemplary carriers, including, for example, saline, polyethylene glycol or glycerol, DPPC, methylcellulose, or mixtures with powder dispersants or fluorocarbons. Aerosol formulations can be placed into pressurized propellants, such as dichlorodifluoromethane, propane, nitrogen, fluorocarbons, and / or other solubilizing or dispersing agents known in the art. Illustratively, delivery can be by using a single-use delivery device, mist nebulizer, breath-activated powder inhaler, aerosol metered-dose inhaler (MDI), or any of the many nebulizer delivery devices available in the art. In addition, mist tents and direct administration through endotracheal tubes can be used.

[0041] In one embodiment, the composition can be formulated, for example, as a solution or suspension, for administration by injection. The solution or suspension can include a suitable non-toxic parenterally acceptable diluent or solvent, such as mannitol, 1,3-butanediol, water, Ringer's solution, or physiological saline, or a suitable dispersing or wetting and suspending agent, such as a sterile, non-irritating, fixed oil, including synthetic mono- or diglycerides, and fatty acids, including oleic acid. One illustrative example of a carrier for intravenous use includes a mixture of 10% USP ethanol, 40% USP propylene glycol or polyethylene glycol 600, and the balance USP water for injection (WFI). Other illustrative carriers for intravenous use include 10% USP ethanol and USP WFI; 0.01-0.1% triethanolamine in USP WFI; or 0.01-0.2% dipalmitoyl diphosphatidylcholine in USP WFI; and 1-10% squalane or a parenteral vegetable oil-in-water emulsion. Water or saline and aqueous dextrose and glycerol solutions may be preferably used as carriers, particularly for injectable solutions. Illustrative carriers for subcutaneous or intramuscular use include phosphate buffered saline (PBS) solutions, 5% dextrose in WFI and 0.01-0.1% triethanolamine in 5% dextrose or 0.9% sodium chloride in USP WFI, or 10% USP ethanol, 40% propylene glycol and 1-2 or 1-4 mixtures of the remainder are acceptable isotonic solutions, such as 5% dextrose or 0.9% sodium chloride, or 0.01-0.2% dipalmitoyl diphosphatidylcholine and 1-10% squalane in USP WFI, or parenteral vegetable oil-in-water emulsions.

[0042] In one embodiment, the adjuvant composition can be formulated for administration via topical application to the skin, for example, as a topical solution, including rinse, spray, drop, lotion, gel, ointment, cream, foam, powder, solid, sponge, tape, steam, paste, tincture, or transdermal patch. Suitable formulations for topical application can include, in addition to any pharma- ceutically active carrier, an emollient, such as carnauba wax, cetyl alcohol, cetyl ester wax, emulsifying wax, hydrous lanolin, lanolin, lanolin alcohol, microcrystalline wax, paraffin, petrolatum, polyethylene glycol, stearic acid, stearyl alcohol, white beeswax, or yellow beeswax. In addition, the composition can include a moisturizer, such as glycerin, propylene glycol, polyethylene glycol, sorbitol solution, and 1,2,6 hexanetriol, or a penetration enhancer, such as ethanol, isopropyl alcohol, or oleic acid.

[0043] As determined by those skilled in the art, additional ingredients such as buffers, carriers, viscosity adjusting agents, preservatives, flavorings, dyes and other ingredients specific to the intended use can be added. Those skilled in the art will recognize that the above description is illustrative rather than exhaustive. Indeed, many additional formulation techniques and pharma- ceutical acceptable excipient and carrier solutions suitable for a particular mode of administration are well known to those skilled in the art.

[0044] In certain embodiments, the dosage ranges from about 0.001 mg / kg body weight to about 1 g / kg body weight. In some embodiments, the dosage ranges from about 0.001 mg / kg body weight to about 0.2 mg / kg body weight, from about 0.001 mg / kg body weight to about 0.1 mg / kg body weight, from about 0.001 mg / kg body weight to about 20 mg / kg body weight, from about 0.001 mg / kg body weight to about 10 mg / kg body weight, from about 0.001 mg / kg body weight to about 5 mg / kg body weight, from about 0.001 mg / kg body weight to about 2 mg / kg body weight, from about 0.001 mg / kg body weight to about 1 mg / kg body weight, from about 0.001 mg / kg body weight to about 0.2 mg / kg body weight, from about 0.001 mg / kg body weight to about 0.02 mg / kg body weight, or from about 0.001 mg / kg body weight to about 0.01 mg / kg body weight. Alternatively, in some embodiments, the dose range is about 0.01 g / kg to about 1 g / kg of body weight, about 0.05 g / kg to about 1 g / kg of body weight, about 0.1 g / kg to about 1 g / kg of body weight, about 0.2 g / kg to about 1 g / kg of body weight, about 0.25 g / kg to about 1 g / kg of body weight, or about 0.5 g / kg to about 1 g / kg of body weight. In one embodiment, the dose range is about 5 μg / kg to about 50 μg / kg of body weight.

[0045] GLP-1RA that reverses aging-related brain symptoms can be given multiple times a day, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 times a day, once a day, less than once a day, once a week, once every two weeks, once a month, once every two months, four times a year, twice a year, once a year, or continuously, to achieve a therapeutically effective dose. The administration of the doses used herein can be repeated for a limited period of time, for example, the doses used herein can be administered daily for several weeks, months or years. The duration of treatment depends on the subject's clinical course and responsiveness to treatment. A therapeutically effective amount is an amount of GLP-1RA sufficient to cause a measurable change in the brain (see "Measurement of Efficacy" below). Such effective amounts can be administered in animal experiments as well as in clinical trials.

[0046] GLP-1RA useful in the present invention can be administered orally, intravenously, intranasally, by inhalation, intraperitoneally, intramuscularly, subcutaneously or intracavity. In one embodiment, the GLP-1RA used herein is administered orally or subcutaneously to the patient.

[0047] Measuring effectiveness In one embodiment, the efficacy of a given treatment can be judged by the improvement of cognitive abilities, as measured by a number of tests, such as memory, recall, visuospatial awareness, speech fluency, expressive language, executive function, gait, and dual tasking, multitasking, etc. These can be reflected as improvements in the scores obtained by cognitive tests, including the Mini-Mental State Examination (MMSE), the Montreal Cognitive Assessment (MoCA) and its variants, the Alzheimer's Disease Assessment Scale-Cognitive subscale (ADAS-Cog) and the Cognitive Dementia Rating (CDR) scale. Clinical improvement is defined as a change in a numerical value in any of the following forms: an increase in MMSE score, an increase in MoCA score, a decrease in ADAS-Cog score, or a decrease in CDR score. Improvement of cognitive abilities can also be reflected as no or slower decline in any of these scores, as compared to an age-matched population, e.g., no change in MMSE, MoCA, ADAS-Cog, or CDR values, which are expected to continue to decline with age.

[0048] In other embodiments, the efficacy of a given treatment can be judged by the reversal of aging-related structural brain changes, which can be measured by imaging methods such as magnetic resonance imaging (MRI), computed tomography (CT) and ultrasonography (US). In certain embodiments, the structural changes can be hippocampal atrophy, cortical atrophy, subcortical structural atrophy, cerebellar atrophy, whole brain atrophy; microbleeds in the gray matter of the frontal, temporal, parietal or occipital lobe; microbleeds in the white matter of the frontal, temporal, parietal or occipital lobe; lacunae, cerebral infarcts, perivascular spaces, white matter intensity changes and / or protein species aggregation in the frontal, temporal, parietal or occipital lobe. Amelioration of structural changes can be in the form of lack of development of new age-related structural changes, static values ​​over time, or altered magnitude of a scale that quantifies the severity of structural changes. The protein species can be amyloid beta, tau, alpha-synuclein, TAR DNA binding protein 43 and / or prion and occur in cortical regions, subcortical regions or the brainstem.

[0049] In other embodiments, the efficacy of a given treatment can be determined by the reversal of functional changes in the brain associated with aging, which changes can be measured using imaging techniques, e.g., functional magnetic resonance imaging (fMRI); magnetic resonance imaging (MRSI); hyperpolarized carbon-13 ( 13 C) Magnetic resonance imaging (MRSI); Ultrasonography (US); Positron Emission Tomography (PET); and Single Photon Emission Computed Tomography (SPECT) can be used to measure brain functional changes. Functional changes in the brain can also be recorded by electrophysiological methods, such as electroencephalography (EEG); intracranial electrode recording; deep brain electrode recording; or magnetic field-based recording methods, such as magnetoencephalography (MEG). Functional changes can be altered brain region activity patterns, neuronal activity, functional connectivity, blood-brain barrier leakage, resting blood flow, glucose consumption, metabolite concentrations, neurovascular coupling, and / or functional hyperemia. For each of these measures, efficacy is defined as a reversal of the respective values ​​close to those found in younger subjects, or a lack of progression of age-related changes in these values ​​that would otherwise occur naturally.

[0050] In other embodiments, the efficacy of a given treatment can be determined by reversal of molecular changes in the brain associated with aging, and the changes can be assessed by measuring molecules from brain tissue or blood, such as gene expression; DNA transcription; RNA translation; location of DNA, RNA or proteins in brain cells or tissues; secretion or release of proteins, DNA, RNA, mitochondria, cellular components or mitochondrial components into the blood. The body fluid can be cerebrospinal fluid (CSF), blood and / or plasma.

[0051] In certain embodiments, calculation of differentially expressed genes (DEGs) associated with the magnitude of change expressed as raw P-value, false discovery rate (FDR) adjusted P-value and / or natural log fold change (lnFC) can be calculated for each cell type. In certain embodiments, DEGs with P-value <0.05, preferably FDR adjusted P-value <0.05.

[0052] In certain embodiments, the genes and subsequent transcribed mRNA and translated proteins that can be assessed in the present invention are the following immune response related genes: synapse modification-related genes, such as Sparcl1, Gpc6, Tgfb2, Megf10, Mertk, Chrdl1; homeostasis function-related genes, such as Kcnj10, Kcnn2, Slc1a2, Slc1a3, Slc6a1, Slc6a9, Slc6a11, Slc7a10, Slc7a11, Slc16a1, Srebf1, Gja1, Gjb6, Itpr2, Grm3, Gria2, Gabbr1, Gabbr2; homeostasis-related genes in MG cells, such as Csf2r and P2ry13; immune activation-related genes in MG cells, such as Ap pe, Ccl3, Ccl4, Cd52, Cst7, Fabp5, Tyrobp, Cd14, Cd33, Ifngr1, Ly86, Map4k4; immune response inhibitory genes in MG cells, such as Cd300a, Il10ra, and Il10rb; calcium signaling related genes in SMCs, such as Camk2g, Stim1, Gsn, Atp2a2, Inpp4b, Mcur1, S100a6, and Tspo; SMC contraction related genes, such as Mylk, Itga1, Mgh11, and Sorbs1; and cell adhesion and ECM remodeling in SMCs, such as Col1a2, Lamb1, Itga7, Jam3, Lamb2, Itgb1, and Bsg.

[0053] Materials and Methods Target animals All experimental methods were approved in advance by the Animal Research Ethics Committee of the Chinese University of Hong Kong (CUHK) and were carried out in accordance with the Guide for the Care and Use of Laboratory Animals. C57BL / 6J mice were provided by the Laboratory Animal Services Centre of CUHK and maintained at a controlled temperature (22–23 °C) with alternating 12-h light / dark cycles, with standard mice having free access to food and water. Ambient humidity was maintained at <70% relative humidity. Male mice of two age groups (2–3 months and 18–20 months of age) were used for the experiments, unless otherwise indicated. For treatment groups, exenatide (5 nmol / kg body weight, Byetta, AstraZeneca LP, Cambridge, UK) or saline vehicle (0.9% w / v sodium chloride) was administered intraperitoneally (IP) (volume: 250 μl / 30 g body weight) daily for 4–5 weeks prior to the experiment, unless otherwise indicated.

[0054] Brain tissue dissociation and single cell isolation A dissociation protocol optimized for single brain cell isolation from both young and old mouse brains (Non-Patent Document 18) was adapted for use. Mice were deeply anesthetized and transcardially perfused with 20 ml of ice-cold phosphate-buffered saline (PBS). The head was then rapidly removed and the whole brain (excluding the cerebellum) was immersed in ice-cold Dulbecco's Modified Eagle Medium (DMEM, Thermo Fisher Scientific, Waltham, MA). Brain tissue was cut into small pieces and dissociated into single cells using a modified version of the Neural Tissue Dissociation Kit (P) (130-092-628, Miltenyi Biotec, Bergisch Gladbach, Germany). Myelin debris was removed using the Myelin Removal kit II (130-096-733, Miltenyi Biotec) according to the manufacturer's manual. Cell clumps were removed by sequential filtration through pre-wetted 70-μm (#352350, Falcon, Corning, NY) and 40-μm (#352340, Falcon) nylon cell strainers. Centrifugation was performed at 300 × g for 5 min at 4 °C. The final cell pellet was resuspended in 500–1000 μl of FACS buffer (phenol red-free DMEM (Thermo Fisher Scientific), supplemented with 2% fetal bovine serum (Thermo Fisher Scientific)).

[0055] Single-cell library generation, sequencing and arraying Single-cell RNA-seq libraries were generated using the Chromium Single Cell 3' Reagent Kit v3 (10X Genomics, Pleasanton, CA). Single-cell suspensions at a density of 500–1000 cells / μL in FACS buffer were added to a real-time polymerase chain reaction (RT-PCR) master mix and loaded onto the Single Cell 3' chip along with Single Cell 3' gel beads and dispensing oil according to the manufacturer's instructions. RNA transcripts derived from single cells were uniquely barcoded and reverse transcribed within the droplets. cDNA molecules were pre-amplified and pooled, and then libraries were constructed according to the manufacturer's instructions. All libraries were quantified by Qubit and RT-PCR on a LightCycler96 system (Roche Life Science, Penzberg, Germany). The size profiles of preamplified cDNA and sequencing libraries were examined by the Agilent High Sensitivity D5000 and High Sensitivity D1000 ScreenTape Systems (Agilent, Santa Clara, CA), respectively. All single-cell libraries were sequenced by customized paired-end in single indexing (28 / 8 / 91-bp for v3 libraries) format as recommended by 10X Genomics. All single-cell libraries were sequenced on a NextSeq 500 system (Illumina, San Diego, CA) using the NextSeq 500 High Output v2 Kit (Illumina). An average of 5897.25 reads per cell (range: 2,340-19,700) were obtained, and an average of 2296.01 genes per cell (range: 88-6,062) were detected. Library sequencing saturation averaged 68.73%. Data were aligned with Cell Ranger (v3.0.0, 10X Genomics).

[0056] Data Quality Management Data processing and visualization were performed using the Seurat package (v3.1.5) and custom scripts in R (v3.6.1). The raw count matrix was generated by default parameters (with the mm10 reference genome). There were 106,832 cells in the primary count matrix. Genes expressed by less than 5 cells were removed, leaving a total of 21,259 genes. Among these genes, 3,000 highly variance genes were identified by the Seurat FindVariableFeatures function. The dataset was filtered to exclude low-quality cells by the following criteria: (1) UMI count or gene count of <5% or >95% or (2) proportion of mitochondrial genes >20%. For dimensionality reduction, principal component analysis (PCA) was applied to calculate the first 30 top principal components. Clustering was performed by the Seurat functions FindNeighbors and FindClusters. The FindNeighbors function constructed a shared nearest neighbor (SNN) graph based on the first 50 principal components. Then, we performed modularity optimization on the clustering SNN results (resolution parameter: 1.2). We visualized the clustering results using Uniform Manifold Approximation and Projection (UMAP).

[0057] Cell type identification To identify the major cell types, we used known cell type-specific marker genes to examine their expression levels in all 52 initial clusters included. We further excluded clusters with double high expression of two or more cell type-specific marker genes. These included clusters with high expression of both endothelial and pericyte markers, corresponding to contamination of pericytes with endothelial cell fragments. We classified the remaining clusters into 16 major cell types (see Figures 1A-1D).

[0058] Expression change gene calculation After quality control and cell type classification, gene count normalization and high variance gene identification were applied to the raw data of 78,490 cells that were retained for further analysis. Using Seurat FindMarkers function and MAST package (v1.8.2), we calculated the differentially expressed genes (DEGs) associated raw P-values, false discovery rate (FDR) adjusted P-values, and the magnitude of the expressed changes in natural logarithm of fold change (lnFC) for each cell type. We defined significant DEGs as those that met an FDR adjusted P-value <0.05.

[0059] Pathway enrichment and disease association analysis We used GeneAnalytics, an online universal gene function analysis tool that contains more than 100 data sources for pathway enrichment analysis. Significant DEGs were semi-automatically converted to human gene orthologs. Pathways with significant functional significance were manually identified, grouped, and summarized from the literature review, including metabolic pathways, immune response and cytokine signaling pathways, respiratory electron transport chain / ATP synthesis and glucose / energy metabolism pathways, gene expression / transcriptional regulation pathways, calcium and other second messenger signaling pathways, cell type-specific pathways (e.g., SMC contraction), hormone signaling pathways, cell adhesion and extracellular matrix remodeling-related pathways.

[0060] Y-maze test Spatial memory was assessed using the Y-maze test. The Y-maze included three enclosed arms made of white acrylic glass, 30 cm long, 8 cm wide, and 15 cm high, set at an angle of 120° to each other. Visual cues were placed around the maze in the test room. This test included two trials. In the first trial, the mouse was allowed to explore the maze for 5 min with one of the arms closed. The mouse was returned to its home cage away from the test room for 2 min between trials. In the second trial, the mouse was allowed to freely explore all three arms of the maze for 5 min. The time spent in each arm was registered from the video recording. Arm entry was defined as the mouse's body crossing the threshold of the central zone of the arm and entering the arm. The percentage of time spent in the novel arm (previously closed in the first trial) was calculated.

[0061] All patents, patent applications, provisional applications and publications mentioned or referenced in this specification are incorporated by reference in their entirety, including all figures and tables, to the extent not inconsistent with the explicit teachings of this specification.

[0062] The following are examples illustrating procedures for carrying out the present invention. These examples are not to be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise stated.

[0063] Working Example Example 1 - Exenatide Treatment of Mice Aging induces dramatic transcriptome changes across multiple major brain cell types, including neurons, glial cells, such as astrocytes (AC), oligodendrocyte precursor cells (OPC), microglia (MG) and oligodendrocytes (OLG), and mural cells, such as endothelial cells (EC), pericytes (PC) and smooth muscle cells (SMC).Treatment with the GLP-1R agonist exenatide reverses these aging-associated transcriptome signatures.The age-associated expression changes reversed by GLP-1RA encompass shared and cell type-specific functional pathways involved in aging and neurodegeneration.

[0064] Single-cell transcriptome profiling was performed in young adult, aged and exenatide-treated aged mice. As shown in Figure 1A-1D, the major cell type clusters identified and analyzed in the mouse brain were visualized by UMAP. Genome-wide expression changes in major cell types in the aging brain and their regulation by GLP-1RA treatment were analyzed. To analyze the pattern and effect of expression changes in aging on exenatide treatment, significant DEGs (defined as false discovery rate (FDR) adjusted P value < 0.05) were calculated for each cell type. As shown in Figure 2A, Figure 2B and Figure 3, the transcriptome changes associated with aging were universally reversed by exenatide treatment across the majority of major cell types in the brain.

[0065] The expression changes reversed by GLP-1RA treatment are functionally relevant. As shown in Figure 4, pathway enrichment analysis for each cell type on the most prominent reversed DEGs highlighted amelioration of age-related expression changes involved in a wide range of cellular functions. In most cell types, these included genes mediating glucose / energy, lipid and protein metabolic processes, as well as transcriptional and translational regulation. There are cell type-specific changes by pathway analysis. In addition, expression changes of some genes play important functional roles. In AC, age-related expression changes of genes mediating immune response, homeostatic function and synaptic plasticity have been reported (Non-Patent Document 3). In this example, AC from GLP-1RA-treated mice appeared to partially revert to a young phenotype by downregulation of several complement 1 component genes (Figure 5) and upregulation of a subset of genes encoding synaptic modification-related proteins, metabolite receptors and transporters, neurotransmitter receptors and ion channels (Figure 5). Meanwhile, immune response and cytokine signaling-related genes were prominent among the DEGs reversed in MG and MAC, followed by EC (Figure 5). Previous studies have reported that microglia in the aging brain exhibit a proinflammatory phenotype (Non-Patent Document 10, Non-Patent Document 13). After GLP-1RA treatment, MG showed upregulation of a number of homeostatic function-related and immune response inhibitory genes, and reversed expression changes of several activation-related genes (Figure 5). In SMC, reversal of expression changes was found after treatment in key functional processes related to age-related vascular sclerosis, including calcium signaling, extracellular matrix (ECM) remodeling, and contractile pathways (Figures 4 and 5).

[0066] Example 2 - Exenatide treatment improves cognitive function in aged mice To evaluate the effect of exenatide to improve cognition in aged mice, 15-17 month old C57BL / 6 were treated daily for 2-4 months with saline vehicle (1 male and 2 female; total 3 mice) or 5 nmol / kg body weight exenatide (4 males and 3 females; total 7 mice). After treatment, the mice were subjected to the Y-maze test. As shown in Figure 6, mice treated with exenatide spent significantly more time in the novel arm, indicating improved spatial memory.

[0067] Example Embodiments Embodiment 1 A method of treating a subject for aging-associated brain dysfunction, comprising administering to the subject an effective amount of a GLP-1R agonist, thereby treating the aging-associated brain dysfunction. Embodiment 2. The method of embodiment 1, wherein the GLP-1R agonist is exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, semaglutide, taspoglutide, PF-06882961, OWL-833, TTP-273, or other molecules that activate GLP-1R. Embodiment 3. The method of embodiment 1, further comprising administering a pharma- ceutically acceptable carrier together with said GLP-1R agonist.

[0023] Embodiment 4. The method of embodiment 1, wherein the subject is a mammal. Embodiment 5. The method of embodiment 4, wherein the mammal is a primate.

[0023] Embodiment 6. The method of embodiment 5, wherein the primate is a human.

[0023] Embodiment 7: The method of embodiment 1, wherein the subject is 50 years of age or older.

[0023] Embodiment 8: The method of embodiment 1, wherein the aging-associated brain dysfunction comprises cognitive impairment. Embodiment 9. The method of embodiment 8, wherein the cognitive impairment is any of attention and concentration, learning tasks and concepts, memory, information processing, visuospatial function, speech, language comprehension, verbal fluency, problem solving, decision-making, and executive function.

[0023] Embodiment 10: The method of embodiment 1, wherein the aging-associated brain dysfunction comprises any of measurable structural, functional and molecular changes in the brain. Embodiment 11 The method of embodiment 10, wherein the measurable structural changes associated with aging are any of hippocampal atrophy, cortical atrophy, subcortical structural atrophy, cerebellar atrophy, whole brain atrophy, cerebral microbleeds in gray matter, cerebral microbleeds in white matter, lacunae, cerebral infarcts, perivascular spaces, white matter intensity changes, and protein species aggregation.

[0023] Embodiment 12. The method of embodiment 11, wherein the protein species is any of amyloid beta, tau, alpha-synuclein, TAR DNA binding protein 43, and prion. Embodiment 13. The method of embodiment 10, wherein the measurable structural changes associated with aging are measured by imaging techniques.

[0046] Embodiment 14: The method of embodiment 13, wherein the imaging method is magnetic resonance imaging (MRI), computed tomography (CT), and / or ultrasound (US). Embodiment 15. The method of embodiment 10, wherein the measurable functional change associated with aging is altered brain regional activity patterns, neuronal activity, functional connectivity, blood-brain barrier leakage, resting blood flow, glucose consumption, metabolite concentrations, neurovascular coupling, or functional hyperemia. Embodiment 16. The method of embodiment 15, wherein the measurable functional changes associated with aging are measured by functional imaging or recording techniques. Embodiment 17 The functional imaging or recording method is functional magnetic resonance imaging (fMRI), magnetic resonance imaging (MRSI), hyperpolarized carbon-13 ( 13 C) The method of embodiment 16, which is magnetic resonance imaging (MRSI), ultrasonography (US), positron emission tomography (PET), single photon emission computed tomography (SPECT), electroencephalography (EEG), magnetoencephalography (MEG), functional near infrared spectroscopy (fNIRS), intracranial electroencephalography, or deep brain electroencephalography. Embodiment 18. The method of embodiment 10, wherein the measurable molecular change associated with aging is a change in gene expression; DNA transcription; RNA translation; location of DNA, RNA or protein in brain cells or tissues; or secretion or release of protein, DNA, RNA, mitochondria, cellular components or mitochondrial components into the blood. Embodiment 19. The method of embodiment 10, wherein the measurable molecular changes associated with aging are changes in cerebrospinal fluid (CSF) or blood / plasma composition.

[0068] It should be understood that the examples and embodiments described herein are for illustrative purposes, and that various modifications or changes in light thereof will be suggested to those skilled in the art that are within the spirit and scope of the present specification and the appended claims. Furthermore, any element or limitation of an invention or embodiment disclosed herein is contemplated within the scope of the present invention, without limitation, with any other element or limitation (individually or in any combination), or with any other invention or embodiment disclosed herein, and all such combinations.

Claims

1. A pharmaceutical composition for reversing and / or inhibiting transcriptomic changes exhibited in cell types in the brain of aged subjects, including mature neurons (mNeur) and immature neurons (imNeur); glial cells astrocytes (AC), oligodendrocyte precursor cells (OPC), microglia (MG), and oligodendrocytes (OLG); mural cells pericytes (PC) and smooth muscle cells (SMC); choroid plexus cells (CPC); hemoglobin-expressing vascular cells (HbEC); and monocytes (MNC), by comparing with a group of young subjects, containing an effective amount of exenatide, The elderly subject is a human aged 50 years or older. Pharmaceutical compositions.

2. 10. The pharmaceutical composition of claim 1, further comprising a pharmaceutically acceptable carrier.

3. The pharmaceutical composition of claim 1 or 2, wherein the elderly subject is a human aged 60 years or older.

4. 2. The pharmaceutical composition of claim 1, A pharmaceutical composition for improving cognitive function associated with aging, including cognitive decline in said elderly subject.

5. The pharmaceutical composition of claim 4 , wherein the cognitive function comprises spatial memory.

6. 2. The pharmaceutical composition of claim 1, wherein the elderly subject has a condition involving measurable structural, functional, or molecular changes in the brain.

7. The pharmaceutical composition of claim 6, wherein the measurable structural change in the brain is hippocampal atrophy.

8. The pharmaceutical composition of claim 6 , wherein the measurable structural change in the brain is measured by an imaging method.

9. 9. The pharmaceutical composition of claim 8, wherein the imaging method is magnetic resonance imaging (MRI), computed tomography (CT), and / or ultrasound (US).

10. The pharmaceutical composition of claim 6, wherein the measurable functional change in the brain is a change in the activity pattern of a region of the brain.

11. The pharmaceutical composition of claim 10, wherein the measurable functional change in the brain is measured by functional imaging or recording methods.

12. The functional imaging or recording method may be functional magnetic resonance imaging (fMRI), magnetic resonance imaging (MRSI), hyperpolarized carbon-13 ( １３ C) The pharmaceutical composition of claim 11, which is magnetic resonance imaging (MRSI), ultrasonography (US), positron emission tomography (PET), single photon emission computed tomography (SPECT), electroencephalography (EEG), magnetoencephalography (MEG), functional near-infrared spectroscopy (fNIRS), intracranial electroencephalography, or deep brain electroencephalography.

13. The pharmaceutical composition of claim 6 , wherein the measurable molecular change in the brain is a change in gene expression.

14. 14. The pharmaceutical composition of claim 13, wherein the measurable molecular changes in the brain are changes in cerebrospinal fluid (CSF) or blood / plasma composition.

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