Lysosomal dysfunction in neurological and psychiatric disorders

Abstract

Compositions for treating lysosomal defects in idiopathic / sporadic brain disorders including lysosomal storage diseases, neuronal ceroid lipofuscinosis, neurological and psychiatric disorders include agents such as troglitazone, rosuvastatin, Compound A, or combinations thereof. TIFF2025503041000016.tif59128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 301,403, filed January 20, 2022, the entirety of which is incorporated by reference herein for all purposes.

[0002] Field The present disclosure relates to a method for treating disease or disorder related to lysosomal dysfunction, which is often accompanied by cognitive changes and neuropsychiatric symptoms.In particular, the composition comprises one or more agents that regulate lysosomal function or activity in disease or disorder, thereby subsequently regulating cognitive and neuropsychiatric symptoms.These include, for example, lysosomal storage disease (LSD), neuronal ceroid lipofuscinosis (NCL), and idiopathic / sporadic neuropsychiatric conditions with lysosomal function deficiency, such as schizophrenia (SZ) and Alzheimer's disease (AD). [Background technology]

[0003] background Idiopathic / sporadic neuropsychiatric disorders have been found to exhibit lysosomal abnormalities. Lysosomes are membrane-bound cellular organelles that are central to the degradative process in animal cells. Extracellular materials, such as microorganisms ingested by phagocytosis, macromolecules by endocytosis, and unwanted cellular organelles, fuse with lysosomes and are degraded to their basic molecules. Thus, lysosomes are the recycling units of the cell. Lysosomes are also responsible for cellular homeostasis for their roles in secretion, cell membrane repair, cell signaling, and energy metabolism.

[0004] Due to the essential role of lysosomes in the cellular degradation process, these organelles are at the crossroads of several cellular processes and have a major impact on health and disease. Defects in one of 60 lysosomal enzymes, transmembrane proteins, or other components of this organelle impede the degradation of target molecules and are responsible for more than 60 different human genetic diseases, collectively known as lysosomal storage diseases. The large number and variety of human pathological conditions characterized by, if not caused by, abnormal lysosomal function highlights the high importance of the autophagy-lysosomal pathway to cellular metabolism. In these diseases and diseases characterized by lysosomal dysfunction, non-degraded material accumulates within lysosomes, which contributes to the presence or severity of the disease, ranging from lysosomal storage diseases to cancer and even cardiovascular diseases. For example, Batten disease (BD) is a group of neuropsychiatric disorders considered the most common neurogenetic NCL, with a prevalence of 1 in 12,500 in some populations. Currently, there are no satisfactory or complete approved treatments for the 14 various forms of Batten disease. Summary of the Invention

[0005] overview In one aspect, the inventors now provide evidence and new concepts that both rare genetic diseases (e.g., BD) and idiopathic / sporadic brain disorders (e.g., SZ, AD) share common lysosomal abnormalities that clearly manifest functional deficits, particularly cognitive dysfunction.

[0006] In certain embodiments, the disease or disorder associated with lysosomal dysfunction includes LSD, NCL, idiopathic / sporadic brain disorders including a wide range of neurological, psychiatric, or functional brain disorders. In certain embodiments, LSDs include glycogen storage diseases, mucopolysaccharidoses, mucolipidoses, oligosaccharidoses, lipidoses, sphingolipidoses, lysosomal trafficking diseases, primary lysosomal hydrolase defects, lysosomal enzyme post-translational processing defects, lysosomal enzyme trafficking defects, lysosomal enzyme protection defects, soluble nonenzymatic lysosomal protein defects, transmembrane (nonenzymatic) protein defects or unclassified defects, Tay-Sachs disease, Sandhoff disease, Niemann-Pick disease, Fabry disease, Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, mucolipidosis type II, mucolipidosis type III, mucopolysaccharidoses, MPS III, MPS VII, GM1 gangliosidosis, and Schindler disease. In certain embodiments, NCL (or BD) includes CLN1 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN5 disease, CLN6 disease, CLN7 disease, CLN8 disease, CLN9 disease, CLN10 disease, CLN11 disease, CLN12 disease, CLN13 disease, and CLN14 disease.

[0007] In certain embodiments, neurological diseases include Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease, prion diseases, amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), Pick's disease, frontotemporal dementia with parkinsonism, multiple sclerosis, Huntington's disease, multiple system atrophy (MSA), Smith-Lemli-Opitz syndrome (SLOS), Tangier disease, Pelizaeus-Merzbach disease, progressive supranuclear palsy, spinal muscular atrophy, and various types of spinocerebellar degeneration and other ataxic conditions. In certain embodiments, psychiatric disorders include schizophrenia, mood disorders, alcoholism and other substance use disorders, autism spectrum disorders, attention deficit hyperactivity disorder, narcolepsy and other sleep disorders, and widespread anxiety disorders.

[0008] In certain aspects, a method for treating a subject who is at risk of or diagnosed with a disease or disorder associated with lysosomal dysfunction is provided, comprising administering a therapeutically effective dose of an agent to the subject, and the agent preferably adjusts lysosomal function or activity, preferably within appropriate homeostatic balance, thereby treating a subject who is at risk of or diagnosed with a disease or disorder associated with lysosomal dysfunction.In certain embodiments, the function or activity of lysosomal in appropriate homeostatic balance is determined by adjusting cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzymes, or combinations thereof.In certain embodiments, the therapeutic effect of the agent is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and combinations thereof.

[0009] In certain preferred aspects, the agent that modulates lysosomal function or activity is an organic small molecule agent.In certain preferred aspects, one or more organic small molecule agents, including one or more organic small molecule agents that can preferably modulate lysosomal function or activity, are administered to a subject to treat the disease or disorder disclosed herein.In one aspect, when the organic small molecule agent shows an effect in an in vitro and / or in vivo model suitable for a particular disorder or disorder disclosed herein, for example, at least about 3, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80 or 100 percent increase in the tested characteristic or parameter relative to a control (e.g., the control can be an assay carried out in the absence of the organic small molecule agent), the agent can be considered as an agent that can modulate lysosomal function or activity.Suitable in vitro and in vivo assays are disclosed herein and / or known.

[0010] In certain aspects, methods and uses are provided for treating the subject suffering from or susceptible to neuronal ceroid lipofuscin (NCL) disease or disorder.In certain preferred aspects, one or more organic small molecule agents, including one or more organic small molecule agents that can preferably regulate the function or activity of lysosome as disclosed herein, are administered to subject to treat neuronal ceroid lipofuscin (NCL) disease or disorder.

[0011] In certain aspects, methods and uses are provided for treating subjects suffering from or susceptible to CLN3 disease.In certain preferred aspects, one or more small organic molecule agents, including one or more small organic molecule agents that can preferably regulate lysosomal function or activity as disclosed herein, are administered to the subject to treat CLN3 disease.

[0012] In certain aspects, methods and uses are provided for treating subjects suffering from or susceptible to Batten disease.In certain preferred aspects, one or more small organic molecule agents are administered to the subject to treat Batten disease, including one or more small organic molecule agents that can preferably modulate lysosomal function or activity as disclosed herein.

[0013] In certain embodiments, the one or more small molecules for administration in the present methods and uses include troglitazone, rosuvastatin, or a combination thereof. A set of small molecules that regulate the overall lipid and protein metabolism, such as troglitazone and rosuvastatin, can directly regulate (particularly improve) lysosomal function.

[0014] In certain embodiments, the one or more small molecules for administration in the present methods and uses may include one or more thiazolidinedione compounds other than or in addition to troglitazone, such as pioglitazone and / or rosiglitazone.

[0015] In certain embodiments, the one or more small molecules for administration in the present methods and uses may include one or more compounds that activate peroxisome proliferator-activated receptors (PPARs). In certain embodiments, the one or more small molecules for administration in the present methods and uses may include one or more compounds that activate PPAR-γ.

[0016] In further embodiments, the one or more small molecules for administration in the present methods and uses may include one or more statin compounds.In particular, the one or more small molecule compounds may include one or more of atorvastatin (Lipitor), fluvastatin (Lescol XL), lovastatin (Altoprev), pitavastatin (Livalo), pravastatin (Pravachol), rosuvastatin (Crestor, Ezallor), and / or simvastatin (Zocor, FloLipid).

[0017] In certain embodiments, the agent comprises other small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments, single-chain antibodies, antibody mimetics, peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, or combinations thereof.In certain embodiments, the agent comprises one or more small molecule compounds.In certain embodiments, the one or more small molecules modulate glycolipid metabolism.In certain embodiments, the method further comprises administering a second therapeutic agent.

[0018] In another aspect, a method for identifying a candidate therapeutic agent (including small organic molecule agents) includes contacting the candidate therapeutic agent with a cell that comprises lysosomal defects, and assaying the modulation of lysosomal function compared to normal control, thereby identifying the candidate therapeutic agent. In a particular embodiment, the candidate therapeutic agent modulates the autofluorescence (AF) of cells, lysosomal size, LAMP1 expression, lysosomal enzymes, and combinations thereof to maintain their homeostatic balance.

[0019] In certain embodiments, the candidate therapeutic agent comprises a small molecule compound, an antisense oligonucleotide, an siRNA reagent, an antibody, an antibody fragment, a single-chain antibody, an antibody mimic, a peptoid, an aptamer, an enzyme, a peptide, an organic or inorganic molecule, a natural or synthetic compound, or a combination thereof. In certain embodiments, the cell comprises a stem cell, an umbilical cord blood cell, an adult stem cell, a mesenchymal stem cell, a mesenchymal stromal cell, an induced pluripotent stem cell (iPSC), an autologous cell, an autologous stem cell, a bone marrow cell, a hematopoietic cell, a hematopoietic stem cell, a somatic cell, a germline cell, a differentiated cell, a somatic stem cell, an embryonic stem cell, a cell line, a patient cell, a mammalian cell, a peripheral blood mononuclear cell (PBMC), a white blood cell, a neuronal cell, a brain cell, or a central nervous system cell. In certain embodiments, the cell comprises an autologous cell, an allogeneic cell, a haploidentical cell, a haploidentical cell, a haploidentical cell, or a xenogeneic cell.

[0020] In one aspect, the small molecule agent (e.g., for use in a method of treating a subject at risk for or diagnosed with a disease or disorder disclosed herein) comprises: The compound has the structure of TIFF2025503041000002.tif35128 or a pharma- ceutically acceptable salt thereof.

[0021] In certain embodiments, the disease or disorder related to lysosomal dysfunction comprises lysosomal storage disease (LSD), neuronal ceroid lipofuscinosis (NCL), idiopathic / sporadic brain disorders, including neurological, psychiatric, or functional brain disorders, and in particular, the disease is Batten disease (BD).In certain embodiments, the NCL or BD comprises CLN3 disease.In certain embodiments, the compound can regulate or modulate the expression of PLA2 gene, particularly PLA2G4A and / or PLA2G7 gene.In certain aspects, the compound can affect the disease pathology that occurs in the hippocampal CA3 region or cerebral cortex in the brain.

[0022] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The following references provide those of ordinary skill in the art with general definitions of many of the terms used in this invention: Academic Press Dictionary of Science and Technology, Morris (Ed.), Academic Press (1 st ed., 1992);Oxford Dictionary of Biochemistry and Molecular Biology, Smith et al. (Eds.), Oxford University Press (revised ed., 2000);Encyclopaedic Dictionary of Chemistry, Kumar (Ed.), Anmol Publications Pvt. Ltd. (2002);Dictionary of Microbiology and Molecular Biology, Singleton et al. (Eds.), John Wiley & Sons (3 rd ed., 2002);Dictionary of Chemistry, Hunt (Ed.), Routledge (1 st Dictionary of Pharmaceutical Medicine, Nahler (Ed.), Springer-Verlag Telos (1994); Dictionary of Organic Chemistry, Kumar and Anandand (Eds.), Anmol Publications Pvt. Ltd. (2002); and A Dictionary of Biology (Oxford Paperback Reference), Martin and Hine (Eds.), Oxford University Press (4 th ed., 2000). Additionally, the following definitions are provided to assist the reader in the practice of the present invention.

[0023] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms thereof unless the context clearly indicates otherwise. Furthermore, to the extent the terms "including," "includes," "having," "has," "with," or derivatives thereof are used in either the detailed description and / or claims, such terms are intended to be inclusive in a similar manner as the term "comprising."

[0024] The term "or" as used herein and in the appended claims is generally used in the sense of including "and / or" unless the context clearly indicates otherwise. Unless specifically stated or clear from the context, the term "about" as used herein is understood to be within the range of general 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 value being referred to. Unless otherwise clear from the context, all numerical values ​​provided herein are modified by the term about.

[0025] As used herein, the terms "administer," "administration," or "administering" refer to (1) the providing, giving, dispensing, and / or prescribing of a composition according to the present disclosure by or under the direction of either a healthcare practitioner or his / her authorized representative, and (2) the patient or person putting into, absorbing, or ingesting a composition according to the present disclosure.

[0026] The term "agent" or "candidate therapeutic agent" as used herein is meant to encompass any molecule, chemical entity, composition, drug, therapeutic, chemotherapeutic, or biological agent that can prevent, ameliorate, or treat lysosomal dysfunction or other medical conditions. The term includes small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments that retain an epitope recognition site, such as Fab, Fab', F(ab')2 fragments, Fv fragments, single chain antibodies, antibody mimetics (e.g., DARPins, affibody molecules, affilins, affitins, anticalins, avimers, fynomers, Kunitz domain peptides, and monobodies), peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, and the like. Agents include compounds that are known drugs, compounds that have been identified as having therapeutic activity but are undergoing further therapeutic evaluation, and compounds that are members of collections and libraries to be screened for pharmacological activity. Agents can be assayed according to the methods of the invention during clinical trials, during preclinical trials, or at any stage after FDA approval.

[0027] As used herein, "cognitive function" can refer to any mental process, including perception, various types of memory, including working memory, emotion control, cognition, attention, reasoning, thinking and judgment abilities, flexibility of thought, and executive function. Measures of cognitive function include, for example, assessment tools designed to measure (a) general intelligence, (b) non-verbal intelligence, (c) achievement, (d) attention / executive function, (e) memory, (f) visual motor and motor function, (g) emotion and anxiety control, (h) reward processes, motivation, and anhedonia scales, (i) cognitive flexibility, set switching and reversal learning, and (j) language.Such assessment tools are well known in the art and include, for example, the Wechsler Adult Intelligence Scale and the Woodcock-Johnson III Test of Cognitive Abilities (both for assessing general intelligence), Raven's Progressive Matrices (for assessing non-verbal intelligence), the Global Achievement Test and the Woodcock-Johnson III Test of Achievement (for assessing academic achievement), Connor's Continuous Performance Test II (for assessing attention / executive function), the Global Assessment of Memory and Learning (for assessing memory and learning), the Bender Visual Motor Gestalt Test, the Halsted-Leitan Grip Strength Test, the Halsted-Leitan Finger Tapping Test, and the Lafayette Grooved Pegboard Task (all for assessing visual motor and motor function), the Cambridge Neuropsychological Test Automated Battery (CANTAB) Emotion Recognition Task, the Pen Computer Neurocognitive Battery (PCNB) Pen Emotion Identification Test, and the Pen Emotion Identification Test, the Mayer-Salovey-Caruso Emotional Intelligence Test (MSCEIT™): a test of emotional intelligence. Emotion Management, Emotion Amplification and Degradation Scales, Emotion Regulation Inventory, Zung Self-Rating Anxiety Scale, Hamilton Anxiety Scale, Beck Anxiety Inventory, Penn Worry State Inventory, Ambiguity Intolerance Scale, Behavioral Inhibition Scale, Negative Evaluation Concern Scale, Anxiety Sensitivity Index, and Life Events and Difficulties Schedule (to assess emotion and anxiety control), Reinforcement Learning, Monetary Incentive Reward, Adaptive Cambridge Gambling, Progressive Ratio, Simple Guessing Task, Fixed Ratio Gratification Schedule, Demotion Task, Habit Task, Habit Learning Task, Probabilistic Reward Task, Pavlovian Conditioning, Drifting Double Bandit, Probability Choice Task, Willingness to Pay Task, Delayed Reward Discounting Task, and Effort-Pay Task (to assess reward processing, motivation, and hedonic and unpleasant scales), CANTAB Intra- and Extra-dimensional Set Shifting, PCNB Penn Conditional Exclusion Test, and Wisconsin Card Sorting Test (to assess cognitive flexibility, set shifting, and reversal learning), and Peabody Picture Vocabulary Test (to assess language). The phrase "improving cognitive function" as referred to herein means a positive change in a subject's ability to perform symbolic operations, such as the ability to perceive, remember, create mental images, have clarity of thought, recognize, reason, think, or judge.As used herein, the term "cognitive decline" refers to a negative change in a subject's ability to perform symbolic operations, such as the ability to perceive, remember, create mental images, have clarity of thought, recognize, reason, think, or judge.

[0028] The term "combination therapy" as used herein refers to a situation in which two or more different agents are administered in an overlapping regimen such that a subject is exposed to both agents at the same time. When used in combination therapy, two or more different agents can be administered simultaneously or separately. This combination administration can include simultaneous administration of two or more agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, two or more agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, two or more agents present in separate formulations can be administered simultaneously. As another alternative, one or more additional agents can be administered immediately after administration of the first agent. In separate administration protocols, two or more agents can be administered minutes apart, hours apart, or days apart.

[0029] The terms "comprising," "comprise," or "including," and their derivatives, as used herein in reference to given or described elements of an article, composition, apparatus, method, process, system, etc., are meant to be inclusive or open-ended, allowing for additional elements, thereby indicating that the given or described article, composition, apparatus, method, process, system, etc. includes the specified element (or its equivalent, where appropriate), and that other elements can be included and still be within the scope / definition of the given article, composition, apparatus, method, process, system, etc.

[0030] The term "contacting" has its normal meaning and refers to combining two or more agents, combining an agent with a cell, or combining two populations of different cells. Contacting can occur in vitro, for example, by mixing a cell with troglitazone and / or rosuvastatin in a test tube or growth medium, or mixing a population of antibodies with a population of cells. Contacting can also occur intracellularly or in situ, for example, by contacting two polypeptides in a cell by co-expression in a cell of a recombinant polynucleotide encoding the two polypeptides, or in a cell lysate. Contacting can also occur in vivo, within a subject or non-human animal, for example, by administering an agent to a subject for delivery of the agent to a target cell.

[0031] The terms "determining," "measuring," "evaluating," "detecting," "assessing," and "assaying" are used interchangeably herein to refer to any form of measurement, including determining whether an element is present or not. These terms include both quantitative and / or qualitative determinations. Evaluating can be relative or absolute. "Assessing the presence of" includes determining the amount of something present, as well as determining whether it is present or absent.

[0032] As used herein, "diagnosis" refers to identifying the presence, extent, and / or nature of a pathological condition. Diagnostic methods differ in their specificity and selectivity. A particular diagnostic method may not provide a definitive diagnosis of a condition, but it is sufficient if the method provides a positive indication that aids in diagnosis.

[0033] As used herein, "lysosomal dysfunction" or "lysosomal defect" refers to any activity, enzymatic or non-enzymatic, or any characteristic of lysosomes that is negatively affected relative to control. This includes vesicular trafficking to or from lysosomes, endocytic pathways, heterophagy or autophagy, and the expression and activity of enzymes localized within lysosomes. "Modulating lysosomal function" refers to increasing or decreasing one or more such activities from the level or amount of such activities found in normal control conditions or within appropriate homeostatic balance. Examples of lysosomal enzymes that can be modulated include lysosomal acid hydrolases, lysosomal proteases, lysosomal nucleases, lysosomal lipases, amylases, and cathepsins. Cathepsin B, cathepsin H or cathepsin L can be assayed using methods known in the art, for example, as described in Barrett, AJ et al., Meth. Enzymol. 80:535 (1981), Academic Press, New York, which is incorporated herein by reference.Lysosomal dysfunction or defect is further described as abnormal lysosomal morphology, chemistry, or activity that is harmful to lysosomes or cells.Examples of lysosomal dysfunction include adverse changes, either increases or decreases in the normal activity of endocytic pathways, adverse changes in lysosomal morphology, intralysosomal pH, and / or adverse changes in the activity of lysosomal enzymes.

[0034] The term "modulator" is used to refer to an entity whose presence in a system in which an activity of interest is observed correlates with a change in the level and / or nature of the activity compared to that observed under otherwise equivalent conditions in the absence of the modulator. In some embodiments, a modulator is an activator, in that the activity increases in its presence compared to that observed under otherwise equivalent conditions in the absence of the modulator. In some embodiments, a modulator is an inhibitor, in that the activity decreases in its presence compared to that observed under otherwise equivalent conditions in the absence of the modulator. In some embodiments, a modulator interacts directly with a target entity whose activity is of interest. In some embodiments, a modulator interacts indirectly with a target entity whose activity is of interest (i.e., directly with an intermediate agent that interacts with the target entity). In some embodiments, a modulator affects the level of a target entity of interest, or alternatively or additionally, in some embodiments, a modulator affects the activity of a target entity of interest without affecting the level of the target entity. In some embodiments, the modulator affects both the level and activity of the target entity of interest such that the differences observed in activity are not fully explained by, and are not to the same extent as, the differences observed in levels.

[0035] As used herein, the terms "nucleic acid," "nucleic acid molecule," "nucleic acid oligomer," "oligonucleotide," "nucleic acid sequence," "nucleic acid fragment," and "polynucleotide" are used interchangeably and are intended to include, but are not limited to, polymeric forms of nucleotides covalently linked together, which may have various lengths, either deoxyribonucleotides and / or ribonucleotides, and / or analogs, derivatives, or modifications thereof. Different polynucleotides may have different three-dimensional structures and may perform various functions, known or unknown. Non-limiting examples of polynucleotides include genomic DNA, genomes, mitochondrial DNA, genes, gene fragments, exons, introns, intergenic DNA (including but not limited to heterochromatic DNA), messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA sequences, isolated RNA sequences, nucleic acid probes, and primers. Polynucleotides useful in the methods of the present disclosure may include naturally occurring nucleic acid sequences and variants thereof, artificial nucleic acid sequences, or combinations of such sequences.

[0036] Nucleic acids, including for example nucleic acids with phosphorothioate backbones, can contain one or more reactive moieties. As used herein, the term reactive moiety includes any group that can react with another molecule, e.g., a nucleic acid or a polypeptide, through covalent, non-covalent, or other interactions. As an example, a nucleic acid can contain an amino acid reactive moiety that reacts with an amino acid on a protein or polypeptide through covalent, non-covalent, or other interactions.

[0037] This term also includes the nucleic acid that contains known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, and which have similar binding properties as reference nucleic acids, and which are metabolized in a similar manner as reference nucleotides.Examples of such analogs include, but are not limited to, phosphoamidate, phosphorodiamidate, phosphorothioate (also known as phosphothioate with double bond sulfur replacing oxygen in phosphate), phosphorodithioate, phosphonocarboxylic acid, phosphonocarboxylate, phosphonoacetic acid, phosphonoformic acid, methylphosphonate, boron phosphonate, or phosphodiester derivatives (see Eckstein, OLIGONUCLEOTIDES AND ANALOGUES: A PRACTICAL APPROACH, Oxford University Press), including nucleotide base modifications, such as in 5-methylcytidine or pseudouridine; and peptide nucleic acid backbones and linkages. Other analog nucleic acids include those with positive backbones, non-ionic backbones, modified sugars, and non-ribose backbones (e.g., phosphorodiamidate morpholino oligos or locked nucleic acids (LNAs) as known in the art), including those described in U.S. Pat. Nos. 5,235,033 and 5,034,506 and ASC Symposium Series 580, CARBOHYDRATE MODIFICATIONS IN ANTISENSE RESEARCH, edited by Sanghui & Cook, Chapters 6 and 7. Nucleic acids containing one or more carbocyclic sugars are also included in one definition of nucleic acid. Modifications of the ribose phosphate backbone can be made for a variety of reasons, for example, to increase the stability and half-life of such molecules in a physiological environment or as a probe on a biochip. Mixtures of naturally occurring nucleic acids and analogs can be made, or mixtures of different nucleic acid analogs and mixtures of naturally occurring nucleic acids and analogs may be made.In embodiments, the internucleotide linkages in the DNA are phosphodiester, phosphodiester derivatives, or a combination of both.

[0038] "Operably linked" refers to a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner. A control sequence "operably linked" to a coding sequence is ligated such that expression of the coding sequence is achieved under conditions compatible with the control sequences.

[0039] A "pharmaceutically acceptable salt" as referred to herein is any salt preparation that is suitable for use in pharmaceutical applications. Pharmaceutically acceptable salts include, but are not limited to, amine salts such as N,N'-dibenzylethylenediamine, chloroprocaine, choline, ammonia, diethanolamine, and other hydroxyalkylamines, ethylenediamine, N-methylglucamine, procaine, N-benzylphenethylamine, 1-para-chloro-benzyl-2-pyrrolidin-1'-ylmethylbenzimidazole, diethylamine, and other alkylamines, piperazine, tris(hydroxymethyl)aminomethane, and the like; alkali metal salts such as lithium, potassium, sodium, and the like; alkaline earth metal salts such as barium, calcium, magnesium, and the like; transition metal salts such as zinc, aluminum, and the like; other metal salts such as sodium hydrogen phosphate, disodium phosphate, and the like; inorganic acids such as hydrochlorides, sulfates, and the like; and salts of organic acids such as acetates, lactates, malates, tartrates, citrates, ascorbates, succinates, butyrates, valerates, fumarates, and the like.

[0040] "Pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers, buffers, etc., such as phosphate buffered saline, 5% aqueous solution of dextrose, and emulsions (e.g., oil / water or water / oil emulsions). Non-limiting examples of excipients include adjuvants, binders, fillers, diluents, disintegrants, emulsifiers, wetting agents, lubricants, flow agents, sweeteners, flavorings, and colorants. Suitable pharmaceutical carriers, excipients, and diluents are described in Remington's Pharmaceutical Sciences, 19th Ed. (Mack Publishing Co., Easton, 1995). The preferred pharmaceutical carrier depends on the intended mode of administration of the active agent. Typical modes of administration include enteral (e.g., oral) or parenteral (e.g., subcutaneous, intramuscular, intravenous, or intraperitoneal injection; or topical, transdermal, or transmucosal administration).

[0041] "Pharmaceutical composition" refers to a composition suitable for pharmaceutical use in a subject animal, including humans and mammals. A pharmaceutical composition comprises a therapeutically effective amount of a therapeutic agent disclosed herein (e.g., an agent that modulates lysosomal function or activity, which in turn modulates cognitive function), optionally another biologically active agent, and optionally a pharmaceutically acceptable excipient, carrier, or diluent. In one embodiment, a pharmaceutical composition includes a composition that includes an active ingredient and an inactive ingredient that constitutes a carrier, as well as any product that results directly or indirectly from the combination, complex formation, or aggregation of any two or more ingredients, or from the dissociation of one or more ingredients, or from other types of reactions or interactions of one or more ingredients. Thus, a pharmaceutical composition of the present disclosure includes any composition that is made by mixing a compound of the present disclosure and a pharmaceutically acceptable excipient, carrier, or diluent.

[0042] The term "promoter" as used herein is defined as a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate specific transcription of a polynucleotide sequence.

[0043] The term "promoter / regulatory sequence" as used herein refers to a nucleic acid sequence required for expression of a gene product operably linked to the promoter / regulatory sequence. In some instances, this sequence may be a core promoter sequence, and in other instances, this sequence may also include enhancer sequences and other regulatory elements required for expression of the gene product. The promoter / regulatory sequence may, for example, be one that causes the gene product to be expressed in a tissue-specific manner.

[0044] A "constitutive" promoter is a nucleotide sequence which, when operably linked to a polynucleotide encoding or specifying a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

[0045] An "inducible" promoter is a nucleotide sequence that, when operably linked to a polynucleotide encoding or specifying a gene product, causes the gene product to be produced in a cell substantially only when an inducer corresponding to the promoter is present in the cell.

[0046] A "tissue-specific" promoter is a nucleotide sequence that, when operably linked to a polynucleotide encoded or specified by a gene, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter.

[0047] A "prophylactic" treatment is a treatment administered to a subject who does not show signs of disease or who shows only early signs of disease, with the intent of reducing the risk of developing a pathology. A compound or composition of the present disclosure may be given as a prophylactic treatment to reduce the likelihood that a pathology will develop, or to minimize the severity of the pathology if it does develop.

[0048] The term "subject" as used herein includes mammals. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates, such as chimpanzees, and other ape and monkey species; livestock animals, such as cows, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. This term does not indicate a particular age or sex. In various embodiments, the subject is a human.

[0049] A "therapeutic" treatment is a treatment administered to a subject exhibiting signs or symptoms of pathology with the intent of diminishing or eliminating those signs or symptoms. The signs or symptoms may be biochemical, cellular, histological, functional or physical, subjective or objective.

[0050] "Treatment" refers to prophylactic or therapeutic treatment. In certain embodiments, "treatment" refers to the administration of a compound or composition to a subject for therapeutic or prophylactic purposes.

[0051] The term "unit dosage form" as used herein refers to a physically discrete unit suitable for unitary administration for human and animal subjects, each unit containing a predetermined amount of a compound of the present disclosure calculated in an amount sufficient to produce a desired effect, optionally in association with a pharma- ceutically acceptable excipient, diluent, carrier, or vehicle. The details of the novel unit dosage forms of the present disclosure depend on the particular compound employed and the effect to be achieved, as well as the pharmacodynamics associated with each compound in the host.

[0052] References herein to "Compound A" refer to the compound having the following structure: TIFF2025503041000003.tif35128 or a pharma- ceutically acceptable salt thereof. [Brief description of the drawings]

[0053] [Figure 1]Figures 1A-1C are a series of plots and electron microscopy images showing that different types of lysosome-associated abnormalities found in cells from patients with schizophrenia (SZ) correlate with cognitive changes. Figure 1A: Cellular autofluorescence (AF) was significantly elevated in peripheral blood cells from SZ patients compared to those from healthy controls (CON). Figure 1B: Immunoelectron microscopy detected altered lysosomes in peripheral blood cells from a subset of SZ patients. We detected changes in the shape and size of this organelle and the accumulation of GAPDH in lysosomes, a sign of stress signals. Green triangles indicate the boundaries of lysosomes, and red triangles indicate GAPDH immunoreactivity. Figure 1C: Levels of AF negatively correlated with cognitive flexibility (Wisconsin Card Sorting Test) scores. CON (white), SZ (black). [Figure 2A] Figures 2A-2D are a series of plots and graphs showing results from the BD cell model. Figure 2A: Correlation between the intensity of autofluorescence (AF) and LAMP1 area in lymphoblasts from healthy subjects (white circles) and juvenile NCL (JNCL) patients (green circles). Figure 2B: Lymphoblasts from JNCL patients showed elevated AF. Troglitazone (3 μM) and rosuvastatin (10 μM) reduced it. Figure 2C: Lymphoblasts from JNCL patients were strongly stained by anti-LAMP1 antibody. Troglitazone (3 μM) and rosuvastatin (10 μM) reduced it (n=3). Figure 2D: Neuronal cells differentiated from Cln3 mutant iPSCs are strongly stained by anti-LAMP1 antibody. Treatment with troglitazone (0.08-2 μM) and rosuvastatin (0.4-10 μM) reduced it in a dose-dependent manner (n=2-3). [Figure 2B] Please see the legend to FIG. 2A. [Figure 2C] Please see the legend to FIG. 2A. [Figure 2D] Please see the legend to FIG. 2A. [Figure 3A]Figures 3A-3C are a series of immunostaining and graphs showing results from a BD animal model (Cln3- / - mice). Figure 3A: Representative images of immunohistochemistry for LAMP1 in brain sections (CA2 / 3 and VPL / VPM) from 6-month-old wild-type mice, untreated and troglitazone (0.06-0.3 mg / g food, for 4 months) treated Cln3- / - mice (left). Quantitative analysis of LAMP1 positive areas in CA2 / 3 and VPL / VPM. Whereas we found accumulation of LAMP1 in untreated Cln3- / - mouse brains, brains from troglitazone-treated mice showed a decrease in LAMP1 positive areas (right, n=8-10). Figure 3B: Novel object recognition (NOR) test using 6-month-old wild-type mice, untreated and troglitazone (0.06-3 mg / g food, for 4 months) treated Cln3- / - mice. Untreated Cln3- / - mice showed a decrease in the NOR test, whereas troglitazone-treated Cln3- / - mice did not. Figure 3C: Peripheral leukocytes from Cln3- / - mice strongly stained with anti-LAMP1 antibody. Troglitazone treatment (0.06-3 mg / g food, for 4 months) reduced it (n=8-10). [Figure 3B] Please see the legend to FIG. 3A. [Figure 3C] Please see the legend to FIG. 3A. [Figure 4A] Figures 4A-4D show the beneficial effects (cytopathology, behavior) of Compound A on Batten-associated pathology in human cells and mouse disease models. A, Compound A ameliorates elevated autofluorescence (AF) in Batten disease (BD) lymphoblasts. B, Compound A ameliorates elevated LAMP1 signal in Batten disease (BD) lymphoblasts. C, Compound A ameliorates elevated LAMP1 signal in peripheral leukocytes of BD mice [CLN3 knockout (KO) mice]. D, Compound A ameliorates cognitive impairment in BD mice (novel object recognition test). [Figure 4B] Please see the legend to FIG. 4A. [Figure 4C] Please see the legend to FIG. 4A. [Figure 4D] Please see the legend to FIG. 4A. [Diagram 5] Figures 5A and 5B show the expression regulation of PLA2G4A (Figure 5A) and PLA2G7 (Figure 5B) by compound A. Compound A significantly affected the expression of PLA2 gene, which may underlie the main mechanism of pathological improvement of Batten CLN3 disease. Data were obtained through RNA sequencing analysis, where we compared gene expression within the following groups: (i) wild-type (WT) neurons + mock, (ii) WT neurons + compound A, (iii) Batten cell model CLN3 knockout (KO) neurons + mock, and (iv) KO neurons + compound A. Neurons were prepared by differentiating human induced pluripotent stem cells. [Figure 6] Figures 6A and 6B show the association between cellular / tissue pathology and behavioral / neuropsychiatric phenotypes by performing histological analysis of autofluorescence (AF). Figure 6A shows that the pathology of Batten CLN3 disease in the brain was prominent in the hippocampal CA3 region, and Figure 6B also shows that the cerebral cortex was involved in CLN3 KO mice. Therefore, the effect of Compound A on the PLA2 gene may be the main mechanism of pathological improvement, in part because the PLA2 gene is directly involved in the metabolism of G3PI. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0054] Detailed Description Idiopathic / sporadic brain disorders have been found to exhibit lysosomal abnormalities. These defects are accentuated, for example, in Alzheimer's disease (AD). Lysosomal defects may be involved in the pathophysiology of psychiatric disorders. For example, alterations in gene expression profiles in postmortem brains from schizophrenia (SZ) patients have been reported. However, investigations of lysosomal defects in psychiatric disorders are still in their infancy. Without wishing to be bound by theory, we hypothesize that lysosomal defects may be a major driving mechanism of neuronal dysfunction.

[0055] There is a group of rare genetic disorders in which lysosomal dysfunction is considered as its central pathology. One representative disorder is BD, also called NCL. BD can be characterized by vision problems, epilepsy, cognitive impairment including emotion / anxiety control, and psychosis. BD patients begin to function during their childhood and can only live for 20-30 years. Mutations in a gene called Cln3 cause about half of the cases of BD, with several other prominent genes such as Cln2. Cln3-driven BD is considered a homogenous condition that exhibits lysosomal pathology. Thus, animal and cell models with disease-associated mutations in Cln3 (or loss of function of Cln3) have been frequently used in research since the gene was identified in 1995. These models are indeed considered effective to investigate lysosomal defects in brain disorders.

[0056] composition In one aspect, the present disclosure provides a composition comprising an agent that regulates glycolipid metabolism as an active ingredient.In certain embodiments, the composition comprises at least one agent that regulates lysosomal function or activity in a therapeutically effective dose.In certain embodiments, the function or activity of lysosomal is determined by regulating cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzyme, or a combination thereof.In certain embodiments, the therapeutic effect of the agent is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and a combination thereof.

[0057] In certain embodiments, the compositions include pharmaceutical compositions comprising troglitazone and rosuvastatin, a set of small molecules that can modulate overall lipid and protein metabolism, which in turn can directly modulate (particularly improve) lysosomal function.

[0058] In certain embodiments, the compositions include pharmaceutical compositions that include other small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments, single chain antibodies, antibody mimetics, peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, or combinations thereof.

[0059] In certain embodiments, the composition comprises a pharmaceutical composition comprising one or more small molecule compounds. In certain embodiments, the one or more small molecules regulate glycolipid metabolism. In certain embodiments, the one or more small molecules comprise troglitazone, rosuvastatin, or a combination thereof.

[0060] As mentioned, in certain embodiments, the one or more small molecules may include one or more thiazolidinedione compounds, such as pioglitazone and / or rosiglitazone, other than or in addition to troglitazone. In certain embodiments, the one or more small molecules may include one or more compounds that activate peroxisome proliferator-activated receptors (PPARs). Suitable troglitazone compounds are also disclosed in US Patent No. 6,207,690.

[0061] In certain embodiments, the one or more small molecules are The compound may include a compound having the structure of TIFF2025503041000004.tif36128 or a pharma- ceutically acceptable salt thereof.

[0062] Compound A can be easily prepared, for example as shown in Scheme 1 below.

[0063] Scheme 1: As shown in step a) of TIFF2025503041000005.tif24147 scheme 1, aryl halide a1 can be reacted under basic conditions to substitute its ring nitrogen with an alkyl group. For example, an electrophile, such as an alkyl halide, can be reacted to provide a substituted ring nitrogen compound, as exemplified by a2 in scheme 1 above. Reagents such as a1 can be commercially available or can be readily prepared, for example, by methods described in Tetrahedron (2007), 63(4), 847-854. As shown in step b) of scheme 1 above, compound a2 is further reacted to provide the desired substituent, as shown for compound A.

[0064] In further embodiments, the one or more small molecules may include one or more statin compounds. In particular, the one or more small molecule compounds may include one or more of atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and / or simvastatin.

[0065] In certain embodiments, the composition comprises a pharmaceutical composition comprising at least one of kaempferide (3,5,7-trihydroxy-4'-methoxyflavone), imiglucerase, velaglucerase, taliglucerase, miglustat, eliglustat, agalsidase beta, agalsidase alpha, migalastat, laronidase, idursulfase, sulfamidase, elosulfase, galsulfase, sebelipase, cerliponase, or combinations thereof.

[0066] In certain embodiments, the compositions include pharmaceutical compositions comprising at least one agent that modulates the expression or activity of phospholipase A2 (PLA2).

[0067] In certain embodiments, the PLA2 includes secreted PLA2 (Groups I, II, III, V, IX, X, XI, XII, XIII, and XIV; e.g., PLA2G1A, PLA2G1B, PLA2G2A, PLA2G2B, PLA2G2C, PLA2G2D, PLA2G2E, PLA2G2F, PLA2G3, PLA2G5, PLA2G9, PLA2G10, PLA2G11A, PLA2G11B, PLA2G12A, PLA2G12B, PLA2G13, and PLA2G14), cytoplasmic PLA2 (Group IV; e.g., PLA2G4A, PLA2G4B, PLA2G4C, PLA2G4D, PLA2G4E, and PLA2G4F), Ca 2+ Independent PLA2s (Group VI; e.g., PLA2G6A, PLA2G6B, PLA2G6C, PLA2G6D, PLA2G6E, and PLA2G6F), platelet-activating factor acetylhydrolases (Groups VII and VIII; e.g., PLA2G7A, PLA2G7B, PLA2G8A, and PLA2G8B), lipoprotein-associated PLA2s (Group XV; e.g., PLA2G15), adipose PLA2s (Group XVI; e.g., PLA2G16) (Dennis et al., Chem Rev. 111:6130 (2011)), or combinations thereof.

[0068] Table 1. Summary of drugs approved for the treatment of lysosomal storage diseases and examples of some products in development that have been granted orphan drug designation. TIFF2025503041000006.tif26159TIFF2025503041000007.tif236159TIFF2025503041000008.tif218159TIFF2025503041000009.tif142149CHO, Chinese hamster ovary cells; MPS, mucopolysaccharidosis; HIRMAb, human insulin receptor monoclonal antibody

[0069] Methods of Treatment and Combination Therapies In certain embodiments, a method of treating a subject at risk for or diagnosed with a disease or disorder associated with lysosomal dysfunction comprises administering to the subject a therapeutically effective dose of an agent (including a small organic molecule agent) that modulates lysosomal function, thereby treating the subject at risk for or diagnosed with a disease or disorder associated with lysosomal dysfunction.

[0070] In certain embodiments, a subject is administered a composition comprising a therapeutically effective dose of at least one agent (including an organic small molecule agent) that modulates lysosomal function or activity. In certain embodiments, lysosomal function or activity is determined by the modulation of cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzymes, or a combination thereof. In certain embodiments, the therapeutic effect of the agent is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and a combination thereof.

[0071] In certain embodiments, a subject is administered a composition comprising a pharmaceutical composition comprising troglitazone and rosuvastatin, a set of small molecules that can regulate overall lipid and protein metabolism, which in turn can directly regulate (particularly improve) lysosomal function.

[0072] In certain embodiments, the subject is administered a composition, including a pharmaceutical composition, that includes other small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments, single chain antibodies, antibody mimetics, peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, or combinations thereof.

[0073] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition comprising at least one of kaempferide (3,5,7-trihydroxy-4'-methoxyflavone), imiglucerase, velaglucerase, taliglucerase, miglustat, eliglustat, agalsidase beta, agalsidase alpha, migalastat, laronidase, idursulfase, sulfamidase, elosulfase, galsulfase, sebelipase, cerliponase, or a combination thereof.

[0074] In certain embodiments, the subject is administered a composition, including a pharmaceutical composition, that includes one or more thiazolidinedione compounds.

[0075] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition that includes one or more thiazolidinedione compounds other than or in addition to troglitazone, such as pioglitazone and / or rosiglitazone.

[0076] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition comprising one or more small molecules for administration in the present methods and uses, which may comprise one or more compounds that activate peroxisome proliferator-activated receptors (PPARs). In certain embodiments, the one or more small molecules for administration in the present methods and uses may comprise one or more compounds that activate PPAR-γ.

[0077] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition comprising one or more statin compounds.In particular, the one or small molecule compound can comprise one or more of atorvastatin (Lipitor), fluvastatin (Lescol XL), lovastatin (Altoprev), pitavastatin (Livalo), pravastatin (Pravachol), rosuvastatin (Crestor, Ezallor), and / or simvastatin (Zocor, FloLipid).

[0078] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition comprising Compound A.

[0079] In certain embodiments, the subject is administered a composition comprising a pharmaceutical composition comprising one or more small molecule compounds.In certain embodiments, the one or more small molecules regulate glycolipid metabolism.In certain embodiments, the one or more small molecules comprise troglitazone, rosuvastatin, or a combination thereof.

[0080] In certain embodiments, one or more therapeutic agents, such as troglitazone, rosuvastatin or a combination thereof, or a statin compound, or compound A, are combined with one or more second therapies or therapeutic agents. These include hematopoietic stem cell transplantation (HSCT), enzyme replacement therapy (ERT), pharmacological chaperone therapy (PCT), substrate synthesis inhibition therapy (SRT), and gene therapy (GT) (Parenti G, et al. New strategies for the treatment of lysosomal storage diseases (Review). Int J Mol Med 31: 11-20, 2013. Michael Beck, Treatment strategies for lysosomal storage disorders. Developmental Medicine & Child Neurology 2018, 60: 13-18).

[0081] Small molecule agents for use in the present therapeutic method can be identified and chemically synthesized using known methods.Small molecule is usually less than about 2000 daltons in size, or alternatively less than about 1500, 750, 500, 250 or 200 daltons in size, and such small molecule can provide the results as disclosed herein in vitro or in vivo (including in vivo model).In this regard, it should be noted that the technology of screening small molecule libraries for molecules that can bind to polypeptide target is well known in the art (see, for example, PCT Publication Nos. WO00 / 00823 and WO00 / 39585). Small molecules can be, for example, fused ring systems (including those containing two, three, or more fused rings and one or more N, O, or S ring atoms, including, for example, compound A disclosed herein), and can contain one or more other functional groups, such as an amine (a primary or, more preferably, a secondary or tertiary alkylamine moiety), an aldehyde, a ketone, an epoxide, or an alcohol.

[0082] Small molecule chemicals can be components of combinatorial chemical libraries. Combinatorial chemical libraries are collections of multiple chemical species composed of smaller subunits or monomers. Combinatorial libraries come in a variety of sizes, ranging from hundreds to hundreds of thousands of different chemical species. There are also a variety of library types, including oligomer and polymer libraries composed of compounds such as carbohydrates, oligonucleotides, and small organic molecules. Such libraries have a variety of uses, such as immobilization and chromatographic separation of chemicals, as well as for identifying and characterizing ligands that can bind to acceptor molecules or mediate biological activities of interest. A variety of techniques are known in the art for synthesizing libraries of compounds on solid supports. Solid supports are typically polymeric objects with a surface functionalized to bind to subunits or monomers to form the compounds of the library. The synthesis of a library typically requires a large number of solid supports. To create a combinatorial library, the solid support is reacted with one or more subunits of the compounds and with one or more many reagents in a carefully controlled, pre-defined sequence of chemical reactions. That is, the library subunits are "grown" on a solid support. The larger the library, the more reactions are required, which complicates the task of keeping track of the chemical composition of the many compounds that make up the library. In some embodiments, the small molecule is less than about 2000 daltons in size, or less than about 1500, 750, 500, 250, or 200 daltons in size.

[0083] Hematopoietic stem cell transplantation: HSCT is based on the use of hematopoietic stem cells from healthy donors as therapeutic agents. This approach can achieve a dual effect; the first is the repopulation of specific tissues with healthy cells of the donor, and the second and key effect is related to the secretion of functional lysosomal hydrolases by the donor's cells in the extracellular space and into the blood circulation. The normal enzymes secreted can be taken up by the recipient cells and cross-correct the enzyme defects in these cells. The great advantage of HSCT is that donor-derived enzyme-producing cells can migrate to the brain. This procedure has the potential to improve neurocognitive function and quality of life, especially if performed early in the disease process.

[0084] Enzyme replacement therapy: ERT is based on regular intravenous infusion of human recombinant lysosomal enzymes, which are produced on a large scale from different sources and purified using recombinant DNA technology. Once injected, the recombinant wild-type enzymes are distributed to tissues, internalized by cells, and targeted to the lysosomal compartment, where they replace the defective enzymes.

[0085] Small molecule pharmacological chaperones: Pharmacological chaperone therapy is based on the concept that loss-of-function diseases are often due to missense mutations that cause misfolding (abnormal conformation) of mutant proteins. Misfolded proteins are recognized and degraded by the quality control system of the endoplasmic reticulum. Thus, in these so-called "misfolded protein diseases", the loss of function is not due to loss of catalytic activity but is the result of the degradation of abnormal proteins. It has been shown that small molecule ligands (pharmacological chaperones) can interact with mutant proteins, support their correct conformation, and improve their stability. As a result, the enzymatic activity of the mutant protein is partially rescued. The use of pharmacological chaperones for the treatment of lysosomal storage diseases was first proposed for Fabry disease. In cells derived from Fabry disease patients, 1-deoxygalactonojirimycin (DGJ), one of the most potent inhibitors of α-gal A, was paradoxically shown to improve the remaining α-galactosidase A activity. In principle, chaperones (as well as substrate reducing drugs and small molecule drugs in general) have several advantages over ERTs, such as the fact that they can be administered orally, thereby allowing for non-invasive treatment, are non-immunogenic, and do not need to be delivered to cells through the mannose-6-phosphate pathway, which may be secondarily impaired in some of these disorders. In addition, small molecule chaperones are expected to diffuse freely through cell membranes to reach therapeutic concentrations in different tissues and systems, including the CNS. M1 In animal models of gangliosidosis, use of the chaperone N-octyl 4-epi-β-valienamine (NOEV) resulted in increased residual activity in the brain and greater clearance of the substrate.

[0086] Proteostasis regulators: Chaperones are ligands that specifically interact with mutant proteins (and thus can rescue single proteins), whereas other drugs can modulate the cellular mechanisms that control protein homeostasis or proteostasis. Proteostasis is a complex network that controls protein synthesis, folding, trafficking, aggregation, and degradation. These small molecule drugs, also known as proteostasis regulators, can modulate the capacity of this network and may restore the normal balance between protein folding, trafficking, and degradation. Two proteostasis regulators have been reported to restore the function of two mutant lysosomal enzymes in two different LSDs: Gaucher disease and GM2 gangliosidosis. Coadministration of a pharmacological chaperone and a proteostasis regulator showed synergistic effects, resulting in further enhancement of enzyme activity. This approach is still largely in the experimental stage.

[0087] Gene Therapy: Gene therapy has the highest potential for complete and lasting correction of enzymatic defects. Gene therapy aims to increase or restore the activity of the defective enzyme in the patient's cells. This is obtained not by supplying the missing enzyme protein, but by delivering a normal copy of the defective gene, which directs the synthesis of the normal enzyme by the recipient's cells. Various viral vectors and various strategies have been investigated to achieve in vivo gene transfer. Herpes viruses, lentiviruses, adeno-associated viruses (AAV), adenoviruses (Ad), etc. have been tested as vectors. Genetic modification can be performed either ex vivo or in vivo. The first strategy is based on ex vivo modification of cells and transplantation of the modified cells into the patient. The cells that are the most widely recognized therapeutic target for LSDs are hematopoietic progenitor cells.

[0088] Substrate Reduction Therapy: SRT is based on the concept that inhibition of certain steps in the biosynthetic pathway of substrates can reduce their entry into lysosomes and help restore the balance between their synthesis and catabolism. This task is generally accomplished by using small molecule inhibitors of enzymes involved in the biosynthesis of the substrates. SRT has already been approved for clinical use for the treatment of Gaucher disease type 1, the most prevalent LSD, caused by β-glucocerebrosidase deficiency and characterized by glycosphingolipid accumulation, and Niemann-Pick disease type C (NPC), a defect in intracellular cholesterol transport.

[0089] CRISPR / Cas9: CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / caspase-9) has the potential to correct specific genetic defects without long-term deleterious side effects. CRISPR / Cas is based on the use of guide ribonucleic acid to guide "molecular scissors" to specific sites in the genome to break double-stranded DNA with the endonuclease caspase-9. The double-stranded DNA break is then repaired by adding donor DNA, ultimately leading to controlled gene modification.

[0090] In certain embodiments, the method of treating a disease or disorder associated with lysosomal dysfunction comprises treating idiopathic / sporadic brain disorders, including LSD, NCL, a wide range of neurological, psychiatric, or functional brain disorders. In certain embodiments, LSDs include glycogen storage diseases, mucopolysaccharidoses, mucolipidoses, oligosaccharidoses, lipidoses, sphingolipidoses, lysosomal trafficking diseases, primary lysosomal hydrolase defects, lysosomal enzyme post-translational processing defects, lysosomal enzyme trafficking defects, lysosomal enzyme protection defects, soluble nonenzymatic lysosomal protein defects, transmembrane (nonenzymatic) protein defects or unclassified defects, Tay-Sachs disease, Sandhoff disease, Niemann-Pick disease, Fabry disease, Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, mucolipidosis type II, mucolipidosis type III, mucopolysaccharidoses, MPS III, MPS VII, GM1 gangliosidosis, and Schindler disease.

[0091] In certain embodiments, NCL (or BD) includes CLN1 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN5 disease, CLN6 disease, CLN7 disease, CLN8 disease, CLN9 disease, CLN10 disease, CLN11 disease, CLN12 disease, CLN13 disease, and CLN14 disease.

[0092] In another embodiment, the methods of treatment are directed to treating Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease, prion diseases, amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), Pick's disease, frontotemporal dementia with parkinsonism, multiple sclerosis, Huntington's disease, multiple system atrophy (MSA), Smith-Lemli-Opitz syndrome (SLOS), Tangier disease, Pelizaeus-Merzbach disease, progressive supranuclear palsy, spinal muscular atrophy, as well as various types of spinocerebellar degeneration and other ataxic conditions.

[0093] In certain embodiments, psychiatric disorders include schizophrenia, mood disorders, alcoholism and other substance use disorders, autism spectrum disorders, attention-deficit hyperactivity disorder, narcolepsy and other sleep disorders, broad-spectrum anxiety disorders.

[0094] In certain embodiments, the treatment of the subject comprises improving cognitive function.In certain embodiments, the treatment of the subject comprises treating cognitive dysfunction or a subset thereof.The examples of cognitive function include, but are not limited to, perception, various types of memory including working memory, emotion control, cognition, attention, reasoning, thinking and judgment ability, flexibility of thought, and executive function.

[0095] In certain embodiments, treating a disease or disorder associated with lysosomal dysfunction or one or more symptoms associated with a disease or disorder associated with lysosomal dysfunction comprises delaying the progression of the disease or one or more symptoms of the disease in time compared to typical disease progression.As used herein, "delaying the progression of a disease or disorder associated with lysosomal dysfunction or one or more symptoms associated with a disease or disorder in time" and the like refers to slowing down and / or stopping the progression of a disease or one or more symptoms of a disease in time (e.g., slowing down and / or stopping the worsening or increasing severity of a disease or one or more symptoms of a disease).Disease progression can be determined, for example, using a known scale, index, grade, or score, such as those described as examples herein, or another suitable test for evaluating progression.For example, the scale, index, grade, score, or other suitable test can correspond to the progression of the entire disease or the progression of one or more symptoms associated with a disease. In one embodiment, the progression of a disease or disorder associated with lysosomal dysfunction or one or more symptoms of a disease or disorder associated with lysosomal dysfunction means that the disease severity value (e.g., overall severity or the severity of one or more symptoms) of a subject determined by known scale, index, grade, score, etc., or other suitable test for evaluating severity does not increase significantly (e.g., at least remains substantially constant).In one embodiment, the progression of a disease or disorder associated with lysosomal dysfunction or one or more symptoms of a disease or disorder associated with lysosomal dysfunction means preventing a subject from reaching a severity value according to known scale, index, grade, score, etc., or other suitable test for evaluating progression compared with the value corresponding to typical disease progression, or increasing the time it takes for a subject to reach it (e.g., reducing the rate of change of severity increase).For example, it can be said that the progression is delayed when the time to reach a severity value is at least 5% longer than that observed according to typical disease progression.Further, for example, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% increase in time is observed. The time that treatment delays the progression of disease or disorder associated with lysosomal dysfunction or one or more symptoms of disease or disorder associated with lysosomal dysfunction can be consistent with the duration of treatment described herein. In one embodiment, treatment delays progression for at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months. Treatment can delay progression for at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years. Treatment can delay progression for the lifetime of the patient.

[0096] For example, to assess the overall neurological condition, the mDRS, a four-domain scale (gait, manipulation, language, and swallowing), may be applied. Cerebellar function may be assessed using the SARA, an eight-item clinical grading scale (gait, posture, sitting, speech, fine motor function, and taxis; ranging from 0 to 40, where 0 is the best neurological condition and 40 is the worst), as well as the SCAFI, which includes the 8m walk time (8MW: performed by having the patient walk from one line to another line twice as fast as possible without turning back), the 9-hole peg test (9HPT), and the number of "PATA" repetitions in 10 seconds. Subjective complaints and quality of life may be assessed using the EQ-5D-5L questionnaire and VAS. To evaluate oculomotor function, three-dimensional video oculography (EyeSeeCam) can be used to measure the peak velocity of saccade, the increase in smooth pursuit, the peak slow phase velocity of gaze-induced nystagmus (gaze-holding function), the peak slow phase velocity of optokinetic nystagmus, and the increase in horizontal vestibulo-ocular reflex.To evaluate cognitive status, WAIS-R or WISC-IV, which evaluate different cognitive domains including attention and concentration, executive function, memory, language, visual construction ability, conceptual thinking, calculation, and orientation, with a maximum score of 30 and a cutoff score of 26, and MoCA can be used.Those skilled in the art are familiar with the methods of carrying out these and other such tests.

[0097] In another embodiment, the identification of biomarkers, expression levels, post-translational modifications, etc. can be used to determine treatment for diseases or disorders related to lysosomal dysfunction.Biomarkers can change over time, which allows medical practitioners to adjust the treatment of subjects.For example, the main CSF biomarkers (T-tau, P-tau, and A1342), CSF NFL, p-tau231, and plasma T-tau are strongly associated with Alzheimer's disease, and these main biomarkers are strongly associated with mild cognitive impairment caused by Alzheimer's disease.

[0098] Candidate Therapeutic Agents In certain embodiments, methods are provided for identifying agents (including small organic molecule agents) with therapeutic activity that may be useful in treating diseases or disorders associated with lysosomal dysfunction. In various embodiments, the screening methods of the present disclosure may be carried out in vitro, in cells, or with transgenic animals. Preferably, cell or animal models for diseases or disorders associated with lysosomal dysfunction, such as BD, SZ, AZ, and other diseases, are used in the methods. The models may also be transgenic animals that exhibit characteristics of diseases or disorders associated with lysosomal dysfunction. Typically, the cell or animal models are contacted with or administered multiple candidate agents. The cell or animal is then tested for cellular responses or phenotypes that demonstrate potential therapeutic activity. As described above, various therapeutic activities may be tested in these methods for dysfunction. For example, these methods may include testing the candidate agents for their ability to modulate lysosomal enzymes. In some embodiments, the candidate agents may be screened for activity by assaying for changes in cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzymes, and combinations thereof.

[0099] In some embodiments, the agent identified in the cell model to have such activity can be further evaluated, for example, in a secondary screen of an animal model of BD, or in a clinical trial to determine activity on movement, behavior, cognition, or other symptoms of the disease.In some related embodiments, the present disclosure also provides a method for evaluating the effect of known drugs and other agents on the treatment of diseases or disorders related to lysosomal dysfunction.In certain embodiments, the candidate agent or potential therapeutic agent modulates some activity or function of lysosome in vivo or in vitro.

[0100] In certain embodiments, the potential therapeutic agents identified based on the screening method are selected to test their therapeutic activity. In certain embodiments, the therapeutic activity of the candidate or potential therapeutic agent is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and combinations thereof.

[0101] In certain embodiments, the therapeutic activity of a candidate or potential therapeutic agent is monitored by PLA2, eg, PLA2G4A or PLA2G7.

[0102] Candidate / Test Agent: A variety of candidate agents, including any naturally occurring or artificially created agent, can be used in the screening methods of the present invention. They can be of any chemical class, such as antibodies, proteins, peptides, small organic compounds, sugars, fatty acids, steroids, purines, pyrimidines, nucleic acids, and various structural analogs or combinations thereof. In some embodiments, the screening methods use combinatorial libraries of candidate agents. Combinatorial libraries can be created for many types of compounds that can be synthesized in a step-by-step manner. Such compounds include polypeptides, beta-turn mimetics, nucleic acids, polysaccharides, phospholipids, hormones, prostaglandins, steroids, aromatic compounds, heterocyclic compounds, benzodiazepines, oligomeric N-substituted glycines, and oligocarbamates. Large combinatorial libraries of compounds can be constructed by the Encoded Synthetic Library (ESL) method described in Affymax, WO 95 / 12608, Affymax, WO 93 / 06121, Columbia University, WO 94 / 08051, Pharmacopeia, WO 95 / 35503, and Scripps, WO 95 / 30642, each of which is incorporated herein by reference for all purposes.

[0103] Candidate agents include many chemical classes, but typically they are organic compounds, including small organic compounds, nucleic acids, including oligonucleotides, peptides, or antibodies. Small organic compounds may suitably have a molecular weight, for example, greater than about 40 or 50, but less than about 2,500. Candidate agents may include functional chemical groups that interact with proteins and / or DNA.

[0104] Candidate agents can be obtained from a wide variety of sources, including libraries of synthetic or natural compounds.For example, numerous means are available for random and directed synthesis of a wide variety of organic compounds and biomolecules, including expression of randomized oligonucleotides.Alternatively, libraries of natural compounds, for example in the form of bacterial, fungal and animal extracts, are available or can be easily produced.

[0105] Chemical Libraries: Developments in combinatorial chemistry have enabled the rapid and economical synthesis of hundreds to thousands of different compounds. These compounds are typically arranged into moderately sized small molecule libraries designed for efficient screening. Combinatorial methods can be used to create unbiased libraries suitable for the identification of novel compounds. In addition, smaller, less diverse libraries may be created that are derived from a single parent compound with previously determined biological activity.

[0106] A combinatorial chemical library is a diverse collection of chemicals that is created by combining a large number of chemical "building blocks", e.g., reagents, either by chemical synthesis or biological synthesis. For example, a linear combinatorial chemical library, e.g., a polypeptide library, is formed by combining a set of chemical building blocks (amino acids) in a large number of combinations and potentially in all possible ways for a given compound length (i.e., the number of amino acids in a polypeptide compound). Millions of chemical compounds can be synthesized through such combinatorial mixing of chemical building blocks.

[0107] A "library" may contain between 2 and 50,000,000 diverse member compounds. Preferably, the library contains at least 48 diverse compounds, preferably 96 or more diverse compounds, more preferably 384 or more diverse compounds, more preferably 10,000 or more diverse compounds, preferably more than 100,000 diverse compounds, and most preferably more than 1,000,000 diverse member compounds. By "diverse" it is meant that more than 50% of the compounds in the library have a chemical structure that is not identical to any other member of the library. Preferably, more than 75%, more preferably more than 90%, and most preferably more than about 99% of the compounds in the library have a chemical structure that is not identical to any other member of the collection.

[0108] The preparation of combinatorial chemical libraries is well known to those of skill in the art. For an overview, see Thompson et al., Synthesis and application of small molecule libraries, Chem Rev 96:555-600, 1996;Kenan et al., Exploring molecular diversity with combinatorial shape libraries, Trends Biochem Sci 19:57-64, 1994;Janda, Tagged versus untagged libraries: methods for the generation and screening of combinatorial chemical libraries, Proc Natl Acad Sci USA. 91: 10779-85, 1994;Lebl et al., One-bead-one-structure combinatorial libraries, Biopolymers 37: 177-98, 1995;Eichler et al., Peptide, peptidomimetic, and organic synthetic combinatorial libraries, Med Res Rev. 15:481-96, 1995;Chabala, Solid-phase combinatorial chemistry and novel tagging methods for identifying leads, Curr Opin Biotechnol. 6:632-9, 1995;Dolle, Discovery of enzyme inhibitors through combinatorial chemistry, Mol. Divers. 2:223-36, 1997;Fauchere et al., Peptide and nonpeptide lead discovery using robotically synthesized soluble libraries, Can J. Physiol Pharmacol.75:683-9, 1997; Eichler et al., Generation and utilization of synthetic combinatorial libraries, Mol Med Today 1: 174-80, 1995; and Kay et al., Identification of enzyme inhibitors from phage-displayed combinatorial peptide libraries, Comb Chem High Throughput Screen 4:535-43, 2001.

[0109] Other chemistries for the creation of chemical diversity libraries can also be used. Such chemistries include peptoids; coded peptides; random bio-oligomers; benzodiazepines (U.S. Pat. No. 5,288,514); diversomers, such as hydantoins, benzodiazepines, and dipeptides; vinylogous polypeptides; non-peptidic peptidomimetics with a β-D-glucose scaffold; analogous organic synthesis of small compound libraries (Chen, et al., J. Amer. Chem. Soc., 116:2661 (1994)); oligocarbamates (Cho, et al., Science, 261:1303 (1993)); and / or peptidyl phosphonates (Campbell, et al., J. Org. Chem. 59:658 (1997)). (1994)); nucleic acid libraries (see Ausubel, Berger, and Sambrook, all supra); peptide nucleic acid libraries (see, e.g., U.S. Pat. No. 5,539,083); antibody libraries (see, e.g., Vaughn, et al., Nature Biotechnology, 14(3):309-314 (1996) and PCT / US96 / 10287); carbohydrate libraries (see, e.g., Liang, et al., Science, 274: 1520-1522 (1996) and U.S. Pat. No. 5,593,853); small organic molecule libraries (e.g., benzodiazepines, Baum C&E News, January 18, p. 33 (1993)); isoprenoids (U.S. Pat. No. 5,569,588); thiazolidinones and metathiazanones (U.S. Pat. No. 5,549,974); pyrrolidines (U.S. Pat. Nos. 5,525,735 and 5,519,134); morpholino compounds (U.S. Pat. No. 5,506,337); benzodiazepines (U.S. Pat. No. 5,288,514), and the like.

[0110] Devices for the preparation of combinatorial libraries are commercially available (see, e.g., 357 MPS, 390 MPS, Advanced Chem. Tech, Louisville Ky.; Symphony, Rainin, Woburn, Mass.; 433A Applied Biosystems, Foster City, Calif.; 9050 Plus, Millipore, Bedford, Mass.). In addition, many combinatorial libraries themselves are commercially available (see, e.g., ComGenex, Princeton, NJ; Asinex, Moscow, Ru; Tripos, Inc., St. Louis, Mo.; ChemStar, Ltd., Moscow, RU; 3D Pharmaceuticals, Exton, Pa.; Martek Bio sciences, Columbia, Md., etc.).

[0111] The screening assay of the present disclosure includes and embodies animal models, cell-based systems, and non-cell-based systems appropriately.The identified gene, its variants, fragments, or oligopeptides are used to identify therapeutically interesting agents, for example by screening a library of compounds, or to otherwise identify compounds of interest by any of a variety of drug screening or analysis techniques.The gene, its alleles, fragments, or oligopeptides used in such screening can be free in solution, fixed to a solid support, produced on the cell surface, or localized intracellularly.Measurements can be carried out as described in detail in the following examples section.

[0112] In some embodiments, methods of identifying candidate therapeutic agents include screening samples containing specific target molecules in a high throughput screening assay.

[0113] In another embodiment, a method for identifying a candidate therapeutic agent for the treatment of a disease includes culturing isolated cells expressing a target molecule, administering a candidate therapeutic agent to the cultured cells, and correlating the expression, activity, and / or function of the target molecule in the presence or absence of the candidate therapeutic agent compared to control cells, and a drug is identified based on a desired therapeutic outcome. For example, a drug that modulates the level of a target molecule, such as a lysosomal enzyme, or a target molecule modulates the activity or amount of another molecule, whether upstream or downstream in a pathway, such that such level is responsible for a disease state. In another example, the assay measures kinase activity. In another example, the assay measures binding partners. In another example, the assay measures the amount of a candidate therapeutic agent that provides a desired therapeutic outcome.

[0114] Another suitable method for diagnosis and drug candidate discovery involves contacting a test sample with cells expressing a target molecule and detecting interaction of a test agent with the target molecule, its alleles or fragments, or an expression product of the target molecule, its alleles or fragments.

[0115] In another preferred embodiment, a sample, e.g., cells or bodily fluids, from a patient is isolated and contacted with a candidate therapeutic molecule, and the genes, their expression products are monitored to identify which genes or expression products are modulated by the drug.

[0116] In another aspect, the present disclosure provides a method for diagnosing or monitoring disease progression in a subject suffering from a disease or disorder associated with lysosomal dysfunction.The therapeutic effect of an agent is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and combinations thereof.These methods include detecting and measuring cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and combinations thereof in a biological sample (e.g., tissue or body fluid sample) from a subject.In some methods, the biological sample is obtained from the brain of a subject.Detecting and quantifying cellular autofluorescence (AF), lysosomal size, and LAMP1 expression in a biological sample can be easily carried out according to the techniques exemplified herein or protocols routinely practiced in the art.

[0117] The present disclosure also provides engineered cells (e.g., neural cells) and transgenic animals that express lysosomal proteins or enzymes. The engineered cells and transgenic animals can be used in vitro or in animal models to study diseases or disorders associated with lysosomal dysfunction as described above, or to test the effects of therapeutic agents. The transgene is preferably present in all or substantially all somatic and germline cells of the transgenic animal. The polynucleotide encoding the full-length and / or mutated and / or truncated lysosomal enzyme is operably linked to one or more regulatory segments that allow the enzyme to be expressed in the cells of the animal. Promoters such as rat neuron-specific enolase promoter, prion protein promoter, human beta-actin gene promoter, human platelet-derived growth factor B (PDGF-B) chain gene promoter, rat sodium channel gene promoter, mouse myelin basic protein gene promoter, human copper-zinc superoxide dismutase gene promoter, and mammalian POU domain regulatory gene promoter can be used to express the transgene. Optionally, an inducible promoter can be used. The mouse metallothionein promoter is suitable, which can be regulated by adding heavy metals such as zinc to the drinking water or food of the mouse.The engineered cells or transgenic animals of the present invention can be produced by general approaches described in the art, for example, Masliah et al., Am. J. Pathol. 148:201-10, 1996; Feany et al., Nature 404:394-8, 2000, and U.S. Patent No. 5,811,633.

[0118] Pharmaceutical Compositions In certain embodiments, pharmaceutical composition comprises at least one agent that regulates lysosomal function or activity.Pharmaceutical composition can comprise agent, such as troglitazone, rosuvastatin, or their pharmaceutically acceptable salt, and pharmaceutically acceptable carrier.Reference to pharmaceutical composition includes active agent alone (e.g., troglitazone, rosuvastatin, or their pharmaceutically acceptable salt, or statin, or compound A) or in the form of pharmaceutical composition.

[0119] Pharmaceutical compositions can be formulated and administered to subjects according to techniques known in the art.Pharmaceutical compositions can take any of many different forms, depending on the manner in which they should be used.Thus, for example, they can be in the form of powder, tablet, capsule, liquid, ointment, cream, gel, hydrogel, aerosol, spray, micelle solution, transdermal patch, liposome suspension, or any other suitable form that can be administered to humans or animals that need treatment.

[0120] As referred to herein, a "pharmaceutical acceptable carrier" is any known compound or combination of known compounds known to those skilled in the art to be useful in formulating a pharmaceutical composition. It will be understood that the carrier of a pharmaceutical composition should be one that is acceptable to the subject to which it is given.

[0121] In one embodiment, the pharma- ceutically acceptable carrier can be solid, and the composition can be in the form of a powder or tablet. A solid pharma- ceutically acceptable carrier can include, but is not limited to, one or more substances that can also function as flavoring agents, buffers, lubricants, stabilizers, solubilizers, suspending agents, wetting agents, emulsifiers, dyes, fillers, glidants, compression aids, inert binders, sweeteners, preservatives, dyes, coating agents, or tablet disintegrating agents. The carrier can also be an encapsulating material. In powder form, the carrier can be a finely divided solid that is in admixture with the finely divided active agent according to the present invention. In tablet form, the active agent can be mixed with a carrier having the required compression properties in an appropriate ratio and compressed into the desired shape and size. Powders and tablets contain, for example, up to 99% of the active agent. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes, and ion exchange resins. In another embodiment, the pharma- ceutically acceptable carrier may be a gel, and the composition may be in the form of a cream, or the like.

[0122] A carrier may include, but is not limited to, one or more excipients or diluents, examples of which are gelatin, gum arabicum, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talcum, colloidal silicon dioxide, etc.

[0123] In another embodiment, the pharma- ceutically acceptable carrier can be liquid. In one embodiment, the pharmaceutical composition is in the form of a solution. Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs, and pressurized compositions. The agent can be dissolved or suspended in a pharma- ceutically acceptable liquid carrier, for example, water, an organic solvent, a mixture of both, or a pharma- ceutical acceptable oil or fat. The liquid carrier can contain other suitable pharmaceutical additives, for example, solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity regulators, solubilizers, or osmolality regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing the above-mentioned additives, for example, cellulose derivatives, for example, sodium carboxymethylcellulose solution), alcohols (including monohydric and polyhydric alcohols, for example, glycols) and their derivatives, and oils (for example, fractionated coconut oil and peanut oil). For parenteral administration, the carrier can also be an oily ester, for example, ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharma- ceutically acceptable propellants.

[0124] Liquid pharmaceutical compositions that are sterile solutions or suspensions can be used, for example, by intramuscular, intrathecal, epidural, intraperitoneal, intravenous, and subcutaneous injection. The active agent can be prepared as a sterile solid composition that can be dissolved or suspended at the time of administration using sterile water, saline, or other appropriate sterile injectable medium.

[0125] The composition may be orally administered in the form of a sterile solution or suspension containing other solutes or suspending agents (e.g., sufficient saline or glucose to make the solution isotonic), bile salts, acacia, gelatin, sorbitan monooleate, polysorbate 80 (oleic acid esters of sorbitol copolymerized with ethylene oxide and its anhydrides), and the like. The composition may also be orally administered in either liquid or solid composition form. Compositions suitable for oral administration include solid forms, such as pills, capsules, granules, tablets, and powders, as well as liquid forms, such as solutions, syrups, elixirs, and suspensions. Forms useful for parenteral administration include sterile solutions, emulsions, and suspensions.

[0126] The composition may alternatively be administered by inhalation (e.g., intranasally). The composition may also be formulated for topical use. For example, a cream or ointment may be applied to the skin.

[0127] The composition can be incorporated into a sustained release or delayed release device. Such a device can be inserted, for example, on or under the skin, and the drug can be released over a period of weeks or even months. Such a device can be advantageous when long-term treatment is required and frequent administration (e.g., at least daily administration) is required.

[0128] In one embodiment, pharmaceutical composition is a solid oral dosage form, such as tablet.In tablet form, active agent can be mixed with a vehicle having necessary compression properties, such as pharmaceutically acceptable carrier, in appropriate ratio, and compressed into desired shape and size.Tablet can contain up to 99% by weight of active agent.

[0129] The pharmaceutical composition, which is a solid oral dosage form, such as a tablet, can be prepared by any method known in the pharmaceutical art. Pharmaceutical compositions are usually prepared by mixing the active agent with a conventional pharma- ceutical acceptable carrier.

[0130] The tablet may be formulated as known in the art, for example, it may contain wheat starch, pregelatinized maize (corn) starch, calcium carbonate, and magnesium stearate as excipients. The same or similar excipients may be used, for example, with the present disclosure.

[0131] As mentioned above, the agent or its pharmaceutically acceptable salt can be formulated and administered as a pharmaceutical composition that can take any of many different forms.For example, the agent or its pharmaceutically acceptable salt can be formulated as a pharmaceutical composition to facilitate its delivery through the blood-brain barrier.As a further example, the agent can be formulated as a pharmaceutical composition to bypass the blood-brain barrier.Formulations that facilitate delivery through the blood-brain barrier or are suitable for administration in a manner that bypasses the blood-brain barrier can be used to prepare and administer the agent as described herein.Such delivery that is suitable for administration through the blood-brain barrier or in a manner that bypasses the blood-brain barrier can be considered as "direct delivery" as used herein.

[0132] In one embodiment, the pharmaceutical composition (e.g., comprising troglitazone, rosuvastatin, or a combination thereof) is formulated for nano-delivery, such as colloidal drug-carrier systems.Suitable examples include, but are not limited to, liposomes, nanoparticles (e.g., polymeric, lipid, and inorganic nanoparticles), nanogels, dendrimers, micelles, nanoemulsions, polymersomes, exosomes, and quantum dots. For example, Patel et al., “Crossing the Blood-Brain Barrier: Recent Advances in Drug Delivery to the Brain,” CNS Drugs 31: 109-133 (2017); Kabanov et al., “New Technologies for Drug Delivery across the Blood Brain Barrier,” Curr Pharm Des., 10(12): 1355-1363 (2004); Cheng et al., “Highly Stabilized Curcumin Nanoparticles Tested in an In Vitro Blood-Brain Barrier Model and in Alzheimer's Disease Tg2576 Mice,” The AAPS Journal, vol. 15, no. 2, pp. 324-336 (2013);Lahde et al. “Production of L-Leucine Nanoparticles under Various Conditions Using an Aerosol Flow Reactor Method,” Journal of Nanomaterials, vol. 2008, article See ID 680897 (2008). For example, direct delivery as used herein can include formulations for nano-delivery.

[0133] In one embodiment, the pharmaceutical composition (e.g., comprising troglitazone, rosuvastatin, or a combination thereof, or a statin, or Compound A) is formulated for direct delivery to the central nervous system (CNS), e.g., by injection or infusion. Formulations and methods for direct delivery to the CNS are known in the art. See, e.g., U.S. Pat. No. 9,283,181. Examples of such administration include, but are not limited to, intranasal, intraventricular, intrathecal, intracranial, administration through nasal mucosa implants, intravenous administration (e.g., IV injection), and oral administration (PO).

[0134] In one embodiment, the pharmaceutical composition is formulated for (and administered by) intranasal delivery. See, for example, Hanson et al., BMC Neurosci. 9(Suppl 3):S5 (2008). In one embodiment, the pharmaceutical composition is formulated for (and administered by) delivery through nasal mucosa implants. In one embodiment, the pharmaceutical composition is formulated for (and administered by) intraventricular injection or infusion. In another embodiment, the pharmaceutical composition is formulated for (and administered by) intrathecal intracisternal injection or infusion. In one embodiment, the pharmaceutical composition is formulated for (and administered by) intrathecal lumbar injection or infusion. For example, the active agent may be formulated for intrathecal administration and / or administered intrathecally in the same or similar manner as described by Ory et al., “Intrathecal 2-hydroxypropyl-beta-cyclodextrin decreases neurological disease progression in Niemann-Pick disease, type C1: a non-randomized, open-label, phase 1-2 trial,” Vol. 390, Issue 10104, pp. 1758-1768 (2017).

[0135] Various techniques can be used, including but not limited to injection through a burr hole or cisterna magna or lumbar puncture, etc., as known in the art. Various devices, such as pumps, catheters, reservoirs, etc., whether internal (e.g., implanted) or external, can be used for delivery, as known in the art. In one embodiment, the administration interval is once every two weeks.

[0136] In yet another embodiment, the pharmaceutical composition is formulated (and administered) for intravenous injection or infusion.For example, Troglitazone, Rosuvastatin, or a combination thereof can be formulated for intravenous administration using needle or tube.In another embodiment, the pharmaceutical composition is formulated (and administered) for oral administration.

[0137] In one embodiment, the administration interval is once a month. In one embodiment, the administration interval is once every two months. In one embodiment, the administration interval is twice a month. In one embodiment, the administration interval is once a week. In one embodiment, the administration interval is twice or several times a week. In one embodiment, the administration interval is daily. In one embodiment, the administration is continuous, for example, continuous infusion.

[0138] The following non-limiting examples are illustrative. EXAMPLES

[0139] Without wishing to be bound by theory, it is hypothesized that lysosomal defects may be a major driving mechanism for neuronal dysfunction.

[0140] Example 1 method Recruitment of Study Participants: Schizophrenia (SZ) patients and age-, sex-, and race-matched controls were recruited at the Johns Hopkins Schizophrenia Center.

[0141] Neurocognitive Testing: A cognitive test battery covering six cognitive domains (cognitive flexibility, attention, processing speed, verbal learning / memory, visual learning / memory, and fluency) was conducted by using our published protocol (Horiuchi et al., Transl Psychiatry, 2016). Cognitive flexibility was assessed using the Wisconsin Card Sorting Test (WCST).

[0142] Electron microscopy of human cells: 70 nm sections of human blood cells were observed on a Hitachi H 7600 TEM operated at 80 kV and digital images were captured using an ER-50 AMT (5 megapixel) CCD camera. Grids without primary antibody served as negative controls. All data were obtained through the Johns Hopkins Core Facility Services.

[0143] Autofluorescence (AF): AF levels in human blood cells were measured in 30,000 cells using flow cytometry at FL-1 on a BD FACSCalibur™ (BD Biosciences) (excitation; 488 nm, emission; 530 ± 15 nm) (Figures 1A-1C). AF levels in lymphoblasts were measured in 20,000 cells using flow cytometry on an Attune Flow Cytometer (Thermo Fisher Scientific) (excitation; 488 nm, emission; 530 nm) (Figures 2A and 2B).

[0144] LAMP1 staining: Lymphoblasts were fixed with methanol and incubated overnight at 4°C with anti-human LAMP1 antibody (1:400, Cell Signaling Technology) followed by secondary antibody (Alexa Fluor 488 donkey anti-rabbit IgG, 1:1000, Thermo Fisher Scientific). Neuronal cells differentiated from iPS cells were fixed with 4% paraformaldehyde (PFA) and incubated overnight at 4°C with anti-human LAMP1 antibody (1:400, Cell Signaling Technology) followed by secondary antibody (Alexa Fluor 488 donkey anti-rabbit IgG, 1:1000, Thermo Fisher Scientific). Leukocytes from mice were fixed with methanol and incubated overnight at 4°C with LAMP1 antibody (1:1000, Abcam) followed by secondary antibody (Alexa Fluor 488 donkey anti-rabbit IgG, 1:1000, Thermo Fisher Scientific). Imaging quantification was performed using a KEYENCE BZ-X710 and BX-Z analyzer software.

[0145] Novel Object Recognition Test (NORT): This test was performed according to a previously validated method (Hatayama et al., Neuroscience 265 (2014) 217-25). All mice were habituated to the NORT arena for 1 h 1 day before the test day. On the test day, mice were given two 5 min trials (T1 with two identical objects and T2 with the familiar object from T1 and a novel object) separated by 1 h. In the T2 trial, the discrimination index (DI) ((time spent exploring the novel object - time spent exploring the familiar object) / total exploration time) was calculated.

[0146] Histological analysis: Mice were sacrificed, brains were excised, and fixation was continued overnight with 4% paraformaldehyde. Brain sections were incubated overnight at 4°C with mouse LAMP1 specific antibody (1:200, Abcam) followed by secondary antibody (Histofine Simple Stain Mouse MAX-PO®, Nichirei Bioscience). For imaging quantification, individual sections were analyzed using Aperio AT-2 and ImageScope (Leica).

[0147] Isolation of leukocytes from peripheral blood of mice: 20 μL of mouse blood was lysed with ACK lysis buffer (Lonza) for 1 min. After washing, cells were resuspended in RPMI.

[0148] RNA sequencing in iPS cell-derived neurons: Cells were differentiated from iPS cells and treated with either 0.1% DMSO or compound A for 2 weeks. RNA was extracted using RNeasy Mini Kit (QIAGEN). Libraries for RNA sequencing were constructed with SMART-Seq v4 Ultra Low Input RNA Kit (Takara bio).

[0149] result The Johns Hopkins Schizophrenia Center has recruited SZ patients over the past decade and conducted extensive tissue collections from the patients and matched controls. One of the goals of this effort was to establish good biomarkers associated with the disease and specific clinical / behavioral manifestations. An interesting finding during such efforts was the discovery that cellular autofluorescence (AF) in peripheral blood cells from SZ patients was elevated compared to those from healthy controls (Figure 1A). Elevated AF has often been reported in BD, and we therefore hypothesized that lysosomal changes could be observed in blood cells from SZ patients. We examined cells at the electron microscopic level and found abnormally enlarged lysosomes (a sign of functional lysosomal defects) in cells from some SZ patients (Figure 1B). SZ includes many clinical manifestations, and we tested which of these distinct clinical aspects were AF-related. Interestingly, AF correlates negatively with cognitive function but not with positive and negative symptoms (Figure 1C). Systematic assessment of multiple cognitive submodalities revealed that cognitive flexibility is a unique modality correlated with AF (Figure 1C).

[0150] Next, we studied BD cell and animal models, especially the Cln3 genetic model. These genetic models recapitulate the lysosomal defects associated with the pathophysiology of BD. The lysosomal defects in lymphoblasts from BD patients with Cln3 mutations were confirmed using two independent measurements: LAMP1 expression and AF levels. We observed a correlation between LAMP1 and AF in this study (Figure 2A). Indeed, these two measurements in a concordant manner confirmed the lysosomal defects (Figures 2B, 2C). Stimulated by the confirmation of lysosomal defects in BD patient cells, we next addressed whether this disease is treatable. To search for promising compounds for treatment in an effective manner, we adopted a hypothesis-based approach, where we hypothesized that chemicals known to affect glycolipid metabolism could directly affect lysosomal function. Accordingly, about 50 compounds were tested in this category. It was found that troglitazone and rosuvastatin could ameliorate the lysosomal defects found in BD lymphoblasts (Figures 2C, 2D). These findings of beneficial effects are novel and not suggested or predicted by anyone. Finally, induced pluripotent stem (iPS) cell lines were tested in which BD-associated Cln3 mutations were introduced. iPS cells were differentiated into neurons, confirming the BD-associated lysosomal defect (Figure 2D). Consistent with the observations in BD lymphoblasts, the treatment effect of troglitazone was also detected in iPS cell-derived neurons carrying the Cln3 mutation (Figure 2D).

[0151] In the postmortem brains of BD patients, there are several distinct brain regions where pathology is more prominent, namely the hippocampus and thalamus. Therefore, we tested for potential pathology in these regions and observed signs of lysosomal defects in tissue sections of the hippocampus and thalamus in one of the most representative BD animal models (Cln3 knockout mice) (Figure 3A). Most importantly, administration of troglitazone ameliorated that pathology (Figure 3A), in agreement with the novel findings in the BD cell model. As described in the introduction section, BD patients show multiple types of neurological and psychiatric signs. Several efforts have been made to address the neurological signs, e.g., seizures, and have shown some good outcomes in BD patient care. However, there is no way to address the cognitive impairment in BD. Therefore, we addressed this through animal models. The first thing we observed was a cognitive impairment in BD animal models in a novel object recognition test (Figure 3B). Therefore, we tested whether troglitazone could ameliorate that deficit and indeed observed a beneficial outcome (Figure 3B). Even at the level of an animal model, this is the first demonstration of a cognitive impairment associated with BD and its amelioration by a small compound. In clinical situations, peripheral biomarkers are always of great importance for the evaluation of disease pathology and treatment response. Therefore, the next question is whether lysosomal defects associated with BD are found in peripheral cells of animal models. Lysosomal defects were tested in peripheral blood cells and defects were observed. Even more importantly, administration of troglitazone in vivo led to the improvement of blood pathology, and such treatment effects on blood cells were consistent with those on neuronal and behavioral changes (Figure 3C).

[0152] summary Through this experiment, we have obtained promising evidence that SZ cells also have lysosomal-related defects. These include elevated AF similar to BD, the defects found in SZ correlate with impairment in distinct cognitive domains, and BD animal models also show lysosomal defects that are believed to underlie cognitive changes. Taken together, it is now logical that both rare genetic diseases (e.g., BD) and idiopathic / sporadic brain disorders (e.g., SZ, AD) may share common lysosomal abnormalities that underlie functional defects. Further experiments will solidify this novel concept. The experimental data also show the promise and possibility that peripheral biomarkers will be established and associated with lysosomal defects, thereby focusing on AF. Finally, the results also show that lysosomal defects and the resulting outcomes, such as behavioral changes, are treatable by small compounds.

[0153] Example 2: Synthesis of Compound A (8-{[2-(dimethylamino)ethyl]amino}-1-(2-methylpropyl)-1,2,3,4-tetrahydro-6H-pyrimido[2,1-b]quinazolin-6-one) Compound A can be synthesized as shown in Scheme 1 below. TIFF2025503041000010.tif31147

[0154] 8-Bromo-1,2,3,4-tetrahydro-6H-pyrimido[2,1-b]quinazolin-6-one (compound a1) is a commercially available product and can also be synthesized according to the method described in Tetrahedron (2007), 63(4), 847-854 or other methods.

[0155] Synthesis of 8-bromo-1-(2-methylpropyl)-1,2,3,4-tetrahydro-6H-pyrimido[2,1-b]quinazolin-6-one (compound a2): 1-bromo-2-methylpropane (3.1 ml) was added dropwise to a mixture of compound a1 (5 g) and cesium carbonate (11.6 g) in DMF (50 ml). After stirring at a temperature of 100° C. for 5 hours, the reaction mixture was quenched with water (300 ml). The resulting solid was collected by filtration and washed with water and hexane. The resulting white solid was dried under reduced pressure at a temperature of 50° C. to obtain compound a1 (5.1 g). TIFF2025503041000011.tif26157

[0156] Synthesis of 8-{[2-(dimethylamino)ethyl]amino}-1-(2-methylpropyl)-1,2,3,4-tetrahydro-6H-pyrimido[2,1-b]quinazolin-6-one (compound A): A solution of compound a2 (450 mg), N,N-dimethylethane-1,2-diamine (139 μl), Brettphos Pd G3 (116 mg), and sodium tert-butoxide (184 mg) in 4-methyltetrahydropyran (15 ml) was stirred at a temperature of 100° C. for 3 hours. Water (50 ml) was added to the reaction mixture, followed by extraction with ethyl acetate. The combined organic layer was dried over MgSO4, filtered, and concentrated. The crude product was added to an amino silica gel column and eluted with hexane / AcOEt followed by AcOEt / MeOH to give the title compound (235 mg) as a yellow solid. TIFF2025503041000012.tif26158

[0157] Example 3 Figures 4A-4D show the beneficial effects of Compound A on Batten-associated pathology (cytopathology, behavior) in human cells and mouse disease models. Figure 4A shows the amelioration of elevated autofluorescence (AF) in Batten disease (BD) lymphoblasts by Compound A. Figure 4B shows the amelioration of elevated LAMP1 signals in Batten disease (BD) lymphoblasts by Compound A. Figure 4C shows the amelioration of elevated LAMP1 signals in peripheral leukocytes of BD mice [CLN3 knockout (KO) mice] by Compound A. Figure 4D shows the amelioration of cognitive impairment in BD mice (novel object recognition test) by Compound A.

[0158] Example 4 Figures 5A and 5B show the expression regulation of PLA2G4A (Figure 5A) and PLA2G7 (Figure 5B) by compound A. Compound A significantly affected the expression of PLA2 gene, which may underlie the main mechanism of pathological improvement of Batten CLN3 disease. Data were obtained through RNA sequencing analysis, in which we compared gene expression between groups (i) wild-type (WT) neurons + mock, (ii) WT neurons + compound A, (iii) Batten cell model CLN3 knockout (KO) neurons + mock, and (iv) KO neurons + compound A. Neurons were prepared by differentiating human induced pluripotent stem cells.

[0159] Example 5 Figures 6A and 6B show the relationship between cell / tissue pathology and behavioral / neuropsychiatric phenotypes by performing histological analysis of autofluorescence (AF). Figure 6A shows that the pathology of Batten CLN3 disease in the brain was prominent in the hippocampal CA3 region, and Figure 6B also shows that the cerebral cortex was involved in CLN3 KO mice. Therefore, the effect of Compound A on the PLA2 gene may be the main mechanism of pathological improvement, in part because the PLA2 gene is directly involved in the metabolism of G3PI.

[0160] Other Aspects From the foregoing, it will be apparent that modifications and variations can be made to the invention described herein to adapt it to various applications and situations. Such embodiments also fall within the scope of the appended claims.

[0161] All citations to sequences, patents, and publications in this specification are herein incorporated by reference to the same extent as if each individual patent and publication was specifically and individually indicated to be incorporated by reference.

Claims

1. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with a disease or disorder associated with lysosomal dysfunction, comprising: The pharmaceutical composition comprises a therapeutically effective dose of one or more agents that modulate lysosomal function or activity.

2. The pharmaceutical composition of claim 1, wherein the function or activity of lysosomes is determined by modulation of cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzymes, or a combination thereof.

3. The pharmaceutical composition of claim 1, wherein the therapeutic effect of the one or more agents is monitored by cellular autofluorescence (AF), lysosomal size, LAMP1 expression, and combinations thereof.

4. 1. A pharmaceutical composition for treating a subject suffering from or susceptible to a lysosomal storage disease (LSD) or neuronal ceroid lipofuscinosis (NCL), comprising: The pharmaceutical composition comprises an effective amount of one or more agents that modulate lysosomal function or activity.

5. 10. The pharmaceutical composition of claim 1 or 4, wherein the one or more agents comprise one or more small organic molecule compounds.

6. 1. A pharmaceutical composition for treating a subject suffering from or susceptible to a lysosomal storage disease (LSD) or neuronal ceroid lipofuscinosis (NCL), comprising: (i) troglitazone and / or rosuvastatin; (ii) one or more peroxisome proliferator-activated receptor (PPAR) activators; (iii) one or more thiazolidinedione compounds; (iv) one or more statin compounds; and / or (v) a structure: or a pharmaceutically acceptable salt thereof, The pharmaceutical composition comprising an effective amount of one or more agents selected from:

7. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise troglitazone and / or rosuvastatin.

8. 7. The pharmaceutical composition of claim 1, 4, or 6, wherein the agent comprises one or more small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments, single-chain antibodies, antibody mimetics, peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, or combinations thereof.

9. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents are small molecules that modulate glycolipid metabolism.

10. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise one or more peroxisome proliferator-activated receptor (PPAR) activators.

11. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise one or more PPAR-γ activators.

12. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise one or more thiazolidinedione compounds.

13. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise pioglitazone and / or rosiglitazone.

14. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise one or more statin compounds.

15. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the one or more agents comprise one or more of atorvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and / or simvastatin.

16. 2. The pharmaceutical composition of claim 1, wherein the disease or disorder associated with lysosomal dysfunction comprises lysosomal storage disease (LSD), neuronal ceroid lipofuscinosis (NCL), idiopathic / sporadic brain disorders including neurological, psychiatric, or functional brain disorders.

17. 17. The pharmaceutical composition of claim 4, 6, or 16, wherein the LSD comprises glycogen storage diseases, mucopolysaccharidoses, mucolipidoses, oligosaccharidoses, lipidoses, sphingolipidoses, lysosomal trafficking diseases, primary lysosomal hydrolase defects, lysosomal enzyme post-translational processing defects, lysosomal enzyme trafficking defects, lysosomal enzyme protection defects, soluble non-enzymatic lysosomal protein defects, transmembrane (non-enzymatic) protein defects or unclassified defects, Tay-Sachs disease, Sandhoff disease, Niemann-Pick disease, Fabry disease, Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, mucolipidosis type II, mucolipidosis type III, mucopolysaccharidoses, MPS III, MPS VII, GM1 gangliosidosis, and Schindler disease.

18. 17. The pharmaceutical composition of claim 4, 6, or 16, wherein the NCL comprises CLN1 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN5 disease, CLN6 disease, CLN7 disease, CLN8 disease, CLN9 disease, CLN10 disease, CLN11 disease, CLN12 disease, CLN13 disease, or CLN14 disease.

19. The object is Alzheimer's disease, Lewy body dementia (LBD), Parkinson's disease, prion diseases, amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), Pick's disease, frontotemporal dementia with parkinsonism, multiple sclerosis, Huntington's disease, multiple system atrophy (MSA), Smith-Lemli-Opitz syndrome (SLOS), Tangier disease, Pelizaeus-Merzbach disease, progressive supranuclear palsy, spinal muscular atrophy, or spinocerebellar degeneration and other ataxic conditions 7. The pharmaceutical composition of claim 1, 4, or 6, wherein the patient is suffering from a neurological disease in which

20. The object is Schizophrenia, mood disorders, alcoholism and other substance use disorders, autism spectrum disorders, attention-deficit hyperactivity disorder, narcolepsy or other sleep disorders, or anxiety disorders 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the patient is suffering from a psychiatric disorder in which:

21. 10. The pharmaceutical composition of claim 1, 4, or 6, wherein the composition is administered to the subject in combination with a second therapeutic agent that is different from the one or more agents.

22. A method for identifying a candidate therapeutic agent, comprising the steps of contacting a candidate therapeutic agent with cells having a lysosomal defect and assaying for modulation of lysosomal function compared to a normal control, thereby identifying the candidate therapeutic agent.

23. 23. The method of claim 22, wherein the candidate therapeutic agent modulates cellular autofluorescence (AF), lysosomal size, LAMP1 expression, lysosomal enzymes, and combinations thereof.

24. 23. The method of claim 22, wherein the candidate therapeutic agent comprises a small molecule compound, an antisense oligonucleotide, an siRNA reagent, an antibody, an antibody fragment, a single-chain antibody, an antibody mimetic, a peptoid, an aptamer, an enzyme, a peptide, an organic or inorganic molecule, a natural or synthetic compound, or a combination thereof.

25. 23. The method of claim 22, wherein the cell comprises a stem cell, an umbilical cord blood cell, an adult stem cell, a mesenchymal stem cell, a mesenchymal stromal cell, an induced pluripotent stem cell (iPSC), an autologous cell, an autologous stem cell, a bone marrow cell, a hematopoietic cell, a hematopoietic stem cell, a somatic cell, a germline cell, a differentiated cell, a somatic stem cell, an embryonic stem cell, a cell line, a patient cell, a mammalian cell, a peripheral blood mononuclear cell (PBMC), a leukocyte, a neuronal cell, a brain cell, or a central nervous system cell.

26. 23. The method of claim 22, wherein the cells comprise autologous cells, allogeneic cells, haplomatched cells, haplomismatched cells, haploidentical cells, or xenogeneic cells.

27. The candidate therapeutic agent identified is (i) in the treatment of a subject at risk for or diagnosed with a disease or disorder associated with lysosomal dysfunction; or (ii) in the treatment of subjects suffering from or susceptible to lysosomal storage diseases (LSDs) or neuronal ceroid lipofuscinosis (NCLs); 23. The method of claim 22, wherein the method is useful.

28. An isolated cell comprising a vector comprising a CLN3 nucleic acid having one or more mutations.

29. 29. The isolated cell of claim 28, wherein the mutation is associated with cognitive impairment or lysosomal defects.

30. 29. The isolated cell of claim 28, wherein the cell comprises a stem cell, an umbilical cord blood cell, an adult stem cell, a mesenchymal stem cell, a mesenchymal stromal cell, an induced pluripotent stem cell (iPSC), an autologous cell, an autologous stem cell, a bone marrow cell, a hematopoietic cell, a hematopoietic stem cell, a somatic cell, a germline cell, a differentiated cell, a somatic stem cell, an embryonic stem cell, a cell line, a patient cell, a mammalian cell, a peripheral blood mononuclear cell (PBMC), a leukocyte, a neuronal cell, a brain cell, or a central nervous system cell.

31. A vector comprising a CLN3 nucleic acid sequence.

32. 32. The vector of claim 31, wherein the CLN3 is operably linked to an inducible promoter, a constitutive promoter, or a tissue-specific promoter.

33. 32. The vector of claim 31, wherein the CLN3 nucleic acid sequence comprises one or more mutations.

34. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with a disease or disorder associated with lysosomal dysfunction, comprising: or a pharmaceutically acceptable salt thereof.

35. 35. The pharmaceutical composition of claim 34, wherein the disease or disorder associated with lysosomal dysfunction comprises lysosomal storage disease (LSD), neuronal ceroid lipofuscinosis (NCL), idiopathic / sporadic brain disorders including neurological, psychiatric, or functional brain disorders.

36. 35. The pharmaceutical composition of claim 34, wherein the disease is Batten disease (BD).

37. 35. The pharmaceutical composition of claim 34, wherein the disease or disorder comprises CLN1 disease, CLN2 disease, CLN3 disease, CLN4 disease, CLN5 disease, CLN6 disease, CLN7 disease, CLN8 disease, CLN9 disease, CLN10 disease, CLN11 disease, CLN12 disease, CLN13 disease, or CLN14 disease.

38. 35. The pharmaceutical composition of claim 34, wherein the subject is suffering from CLN3 disease.

39. 35. The pharmaceutical composition of claim 34, wherein the disease pathology occurs in the hippocampal CA3 region or the cerebral cortex of the brain.

40. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with an additional agent comprising one or more small molecule compounds, antisense oligonucleotides, siRNA reagents, antibodies, antibody fragments, single-chain antibodies, antibody mimetics, peptoids, aptamers, enzymes, peptides, organic or inorganic molecules, natural or synthetic compounds, or combinations thereof.

41. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with troglitazone, rosuvastatin, or a combination thereof.

42. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with the one or more peroxisome proliferator-activated receptor (PPAR) activators.

43. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with one or more PPAR-γ activators.

44. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with one or more thiazolidinedione compounds.

45. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with pioglitazone and / or rosiglitazone.

46. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with one or more statin compounds.

47. 35. The pharmaceutical composition of claim 34, wherein the composition is administered to the subject in combination with one or more of atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and / or simvastatin.

48. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: or a pharmaceutically acceptable salt thereof.

49. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: The pharmaceutical composition comprises a therapeutically effective amount of troglitazone, rosuvastatin, or a combination thereof.

50. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: The pharmaceutical composition comprises a therapeutically effective amount of one or more peroxisome proliferator-activated receptor (PPAR) activators.

51. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: The pharmaceutical composition comprises a therapeutically effective amount of one or more thiazolidinedione compounds.

52. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: The pharmaceutical composition comprises a therapeutically effective amount of one or more statin compounds.

53. 53. The pharmaceutical composition of claim 52, wherein the one or more statin compounds comprise one or more of atorvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, and / or simvastatin.

54. 1. A pharmaceutical composition for treating a subject at risk for or diagnosed with Batten disease, comprising: The pharmaceutical composition comprises a therapeutically effective amount of one or more small molecule compounds capable of modulating lysosomal function or activity.

55. A pharmaceutical composition for treating a subject at risk for or diagnosed with a disease or disorder disclosed herein, comprising: The pharmaceutical composition comprises a therapeutically effective amount of one or more small molecule compounds capable of modulating lysosomal function or activity.

56. A pharmaceutical composition for treating a subject suffering from or susceptible to a disease or disorder disclosed herein, comprising: (i) troglitazone and / or rosuvastatin; (ii) one or more peroxisome proliferator-activated receptor (PPAR) activators; (iii) one or more thiazolidinedione compounds; (iv) one or more statin compounds; and / or (v) a structure: or a pharmaceutically acceptable salt thereof, The pharmaceutical composition comprising an effective amount of one or more agents selected from:

57. A compound having the structure: or a pharmaceutically acceptable salt thereof.

58. 58. A pharmaceutical composition comprising the compound of claim 57, or a pharmaceutically acceptable salt thereof, and a carrier.

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