Compositions and methods to treat neurodegeneration and cognitive decline

Abstract

The present disclosure provides methods for treating neurodegeneration or cognitive decline in a subject comprising administering an effective amount of catestatin (CST) or its equivalent to a subject afflicted with a neurodegenerative disease, thereby, reducing neurodegeneration or cognitive decline.

Description

Atty. Dkt. No.: 114198-3450COMPOSITIONS AND METHODS TO TREAT NEURODEGENERATION AND COGNITIVE DECLINESTATEMENT OF GOVERNMENT SUPPORT

[0001] This invention was made with government support under AG080246 and AG078635 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0002] Neurodegeneration refers to the progressive loss of structure or function of neurons, including their death. This process is a hallmark of various neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease. These conditions gradually damage and destroy parts of the nervous system, particularly the brain, leading to a decline in cognitive functioning.

[0003] Cognitive decline is a common symptom of neurodegenerative diseases and can manifest in various ways, including memory loss, difficulty with problem-solving, and impaired judgment. Better treatments for slowing down or reversing neurodegeneration and cognitive decline are needed.

[0004] Tauopathies are a broad class of neurodegenerative diseases characterized by the accumulation of tau aggregates in the brain. Primary tauopathies such as Pick’s disease (PiD: 3R, where R represents microtubule binding domain), corticobasal degeneration (CBD: 4R), progressive supranuclear palsy (PSP: 4R), globular glial tauopathy (GGT: 4R), argyrophilic grain disease (AGD: 4R) and exhibit only tau inclusions, whereas AD, a secondary tauopathy, is characterized by the presence of extracellular beta-amyloid (AP) plaques and intracellular neurofibrillary tau tangles. Six major isoforms were detected in the brains of AD patients: 2N4R (441 aa), 1N4R (412 aa), 0N4R (383 aa), 2N3R (410 aa), 1N3R (381 aa) and 0N34 (352aa), where N represents the amino-terminal region. Hyperphosphorylation of tau weakens tau’s microtubule-binding affinity, leading to its neurotoxic aggregation, forming tangles of different substructures. Tauopathies are closely associated with metabolic dysfunctions, including insulin resistance, hyperglycemia, and hypertension, as well as neuroinflammation. However, the pathophysiology underlying tauopathy development during aging remains elusive.14867-5418-2384.2Atty. Dkt. No.: 114198-3450SUMMARY OF THE DISCLOSURE

[0005] In one aspect, provided herein is a method for treating neurodegeneration in a subject in need thereof comprising, or alternatively consisting essentially of, or consisting of, or yet further consisting of administering an effective amount of chromogranin A (CgA)-derived peptide catestatin (CST: human CgA352-372) or an equivalent thereof to the subject.

[0006] In some embodiments, the effective amount of CST is an amount that results in (i) at least 50% decrease in phosphorylation of Tau in the subject or (ii) an improvement in memory function of the subject.

[0007] In some embodiments, the effective amount is between about 20mg and about 200mg.

[0008] In some embodiments, the subject in need thereof has low levels of naturally occurring CST in the subject’s plasma, cortex or hippocampus.

[0009] In some embodiments, the subject is suffering from a tauopathy.

[0010] In some embodiments, the CST or the equivalent thereof comprises, or alternatively consists essentially of, or consists of, or yet further consists of a peptide selected from SEQ ID NOs: 1-28.

[0011] In some embodiments, the CST or the equivalent thereof further comprises a nonnatural amino acid at position R8 of SEQ ID NO: 1, or an equivalent thereof wherein the non- naturally amino acid at position R8 is retained.

[0012] In some embodiments, the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons.

[0013] In some embodiments, the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety.

[0014] In some embodiments, the equivalent of CST comprises, or alternatively consists essentially of, or consists of, or yet further consists of a retro-inverso peptide of any of the sequences selected from SEQ ID NOs: 1-28

[0015] In some embodiments, the CST or an equivalent thereof is administered every 12 hours, once every day, once every three days, once every week, once in two weeks or once every month.

[0016] In some embodiments, the subject is a mammal, optionally a human patient.24867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0017] In some embodiments, the CST or an equivalent thereof is administered by an enteral route, buccal route, intraperitoneal route, inhalation route, intravenous route, subcutaneous route or intramuscular route.BRIEF DESCRIPTION OF THE FIGURES

[0018] FIGS. 1A-1I. Biochemical determination of CST in human samples. CST content was found to have decreased (A) a -36% decrease in AD frontal cortex, (B) a -19% decrease AD hippocampus, (C) a -29% decrease in CBD frontal cortex, and (D) a -37% decrease in PSP basal ganglia. AD patients were diagnosed as having Braak stage 0-2 or Braak stage 6. Braak stages I and II are used when neurofibrillary tangle involvement is confined mainly to the transentorhinal region of the brain, stages III and IV when there is also involvement of limbic regions such as the hippocampus, and V and VI when there is extensive neocortical involvement. CBD and PSP patients were diagnosed as having Braak stage 0-1 or Braak stage 2-3. (E) Phospho-Tau in AD frontal cortex. Applicant used mini -mental state examination score (MMSE) values to determine the disease severity (impairment in mental health): a score of >25 is considered normal; a score of 21-24 is considered mild impairment; a score of 11-20 is considered moderate impairment; a score of <10 is considered severe. To correlate CST concentration and disease severity, Applicant ran a simple linear regression between MMSE score and CST concentration in brain. Applicant found that a low concentration of CST is associated with the worst prognosis in both AD cortex (F) and hippocampus (G). Western blot analyses revealed phosphorylation of Tau in Braak stage 6 in the AD frontal cortex (E) and Braak stage 3 in the CBD frontal cortex (I) compared to no phosphorylation in Braak stage 0. Interestingly, CST concentration is inversely correlated with phospho-Tau (S202 / T205) (H), indicating an association of CST with Tau phosphorylation and aggregation. **p<0.01; ***p<0.001; ****p<0.0001.

[0019] FIGS. 2A-2C. CST content in human and PS19-Tg mice. (A) a -47% decrease in CST concentration in CSF of AD patients. (B) a -33% decrease in CST content in PS 19 mouse cortex as compared to WT. (C) Age-dependent decrease (-29-39%) in plasma CST concentration in PS 19 mice. PS 19 mice are a model to study tau pathology, tau aggregates, as well as other AD-related symptoms such as age-associated cognitive impairment. PS 19 mice are overexpressing a human tau T34 isoform and 4 microtubule binding repeats (1N4R) of the tau protein with P301S mutation under the regulatory control of the murine prion promoter. *p<0.05; **p<0.01; ***p<0.001.34867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0020] FIGS. 3A-3D. CST decreased Tau phosphorylation and seeding in organotypic slice culture. (A) Schematic for organotypic slice culture generation, treatment and imaging. (B) Immunoblots showing phosphorylated and total Tau. (C) Tau aggregate staining. (D) Semi -quantitation of B&C showing a -59% decrease in Tau aggregation in response to CST. *p<0.05.

[0021] FIGS. 4A-4D. CST caused up to -73% decrease in Tau phosphorylation in vivo in PS19-Tg mice. (A) Cartoon showing chronic treatment plan used for this study. (B) Immunoblots showing phosphorylated and total Tau after 4 months of treatment with CST (0.5 pg / g body weight; 3 days a week). (C&D) Densitometric analysis of immunoblots in (A) showing a -73% decrease in phosphorylation of Tau at S202 / T205 and S396 / S404 in response to CST, indicating CST regulation of Tau proteinopathy. ***p<0.001;****p<0.0001. BW, body weight; M, Monday; W, Wednesday; F, Friday.

[0022] FIGS. 5A-D. CST caused up to -76% decrease Tau aggregation in vivo in PS19-Tg mice. MCI staining was significantly decreased in CA3 (A) and dentate gyrus (C) regions after 4 months of CST treatment. Semiquantative evaluation revealed a -76% decrease in Tau aggregation in both CA3 (B) and dentate gyrus (D) regions.DETAILED DESCRIPTIONDefinitions

[0023] As it would be understood, the section or subsection headings as used herein is for organizational purposes only and are not to be construed as limiting and / or separating the subject matter described.

[0024] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods, devices, and materials are now described. All technical and patent publications cited herein are incorporated herein by reference in their entirety. Nothing herein is to be construed as an admission that the disclosure is not entitled to antedate such disclosure by virtue of prior disclosure.

[0025] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell44867-5418-2384.2Aty. Dkt. No.: 114198-3450 biology and recombinant DNA, which are within the skill of the art. See, e.g., Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; the series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., N.Y.); MacPherson et al. (1991) PCR 1 : A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition; Gait ed. (1984) Oligonucleotide Synthesis; U.S. Patent No. 4,683,195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds. (1984) Transcription and Translation; Immobilized Cells and Enzymes (IRL Press (1986); Perbal (1984) A Practical Guide to Molecular Cloning; Miller and Calos eds. (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London);Herzenberg et al. eds (1996) Weir’s Handbook of Experimental Immunology; Manipulating the Mouse Embryo: A Laboratory Manual, 3rd edition (Cold Spring Harbor Laboratory Press (2002)); Sohail (ed.) (2004) Gene Silencing by RNA Interference: Technology and Application (CRC Press).

[0026] As used in the specification and claims, the singular form “a,” “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a cell” includes a plurality of cells, including mixtures thereof.

[0027] As used herein, the term “comprising” is intended to mean that the compounds, agents, compositions and methods include the recited elements, but not exclude others. “Consisting essentially of’ when used to define compounds, agents, compositions and methods, shall mean excluding other elements of any essential significance to the combination. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants, e.g., from the isolation and purification method and pharmaceutically acceptable carriers, preservatives, and the like. “Consisting of’ shall mean excluding more than trace elements of other ingredients. Embodiments defined by each of these transition terms are within the scope of this technology.

[0028] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which are varied (+) or (-) by increments of 1, 5, or 10%. It is to be understood, although not always explicitly stated that all numerical54867-5418-2384.2Aty. Dkt. No.: 114198-3450 designations are preceded by the term “about.” It also is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0029] The term “about,” as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1 % of the specified amount.

[0030] As used herein, comparative terms as used herein, such as high, low, increase, decrease, reduce, or any grammatical variation thereof, can refer to certain variation from the reference. In some embodiments, such variation can refer to about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 1 fold, or about 2 folds, or about 3 folds, or about 4 folds, or about 5 folds, or about 6 folds, or about 7 folds, or about 8 folds, or about 9 folds, or about 10 folds, or about 20 folds, or about 30 folds, or about 40 folds, or about 50 folds, or about 60 folds, or about 70 folds, or about 80 folds, or about 90 folds, or about 100 folds or more higher than the reference. In some embodiments, such variation can refer to about 1%, or about 2%, or about 3%, or about 4%, or about 5%, or about 6%, or about 7%, or about 8%, or about 0%, or about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 75%, or about 80%, or about 85%, or about 90%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99% of the reference.

[0031] As will be understood by one skilled in the art, for any and all purposes, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Furthermore, as will be understood by one skilled in the art, a range includes each individual member.

[0032] “Optional” or “optionally” means that the subsequently described circumstance may or may not occur, so that the description includes instances where the circumstance occurs and instances where it does not.

[0033] As used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).

[0034] “Substantially” or “essentially” means nearly totally or completely, for instance, 95% or greater of some given quantity. In some embodiments, “substantially” or “essentially” means 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9%.64867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0035] The terms or “acceptable,” “effective,” or “sufficient” when used to describe the selection of any components, ranges, dose forms, etc. disclosed herein intend that said component, range, dose form, etc. is suitable for the disclosed purpose.

[0036] A “composition” is intended to mean a combination of active agent and another compound or composition, inert (for example, a detectable agent or label) or active, such as an adjuvant, diluent, binder, stabilizer, buffers, salts, lipophilic solvents, preservative, adjuvant or the like and include pharmaceutically acceptable carriers.

[0037] Carriers also include pharmaceutical excipients and additives proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri, tetraoligosaccharides, and oligosaccharides; derivatized sugars such as alditols, aldonic acids, esterified sugars and the like; and polysaccharides or sugar polymers), which can be present singly or in combination, comprising alone or in combination 1-99.99% by weight or volume. Exemplary protein excipients include serum albumin such as human serum albumin (EISA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acid components, which can also function in a buffering capacity, include alanine, arginine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. Carbohydrate excipients are also intended within the scope of this technology, examples of which include but are not limited to monosaccharides such as fructose, maltose, galactose, glucose, D- mannose, sorbose, and the like; disaccharides, such as lactose, sucrose, trehalose, cellobiose, and the like; polysaccharides, such as raffinose, melezitose, maltodextrins, dextrans, starches, and the like; and alditols, such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol) and myoinositol.

[0038] Introduction of non-natural amino acids into CST sequence. Substitution of natural amino acids by the non-natural amino acids improves the pharmacokinetic properties of bioactive peptides. Since plasmin and cathepsin L cleaves CST at R8, Applicant replaces R8 with D-Arginine, a methyl amino acid or a P-amino acid.

[0039] Conjugation of CST with fatty acids. Fatty acids of different lengths, such as CIO (e.g., capric acid), C12 (e.g., lauric acid), C14 (e.g., hexadecenoic acd), C16 (e.g., palmitic acid), or Cl 8 (e.g., linoeic acid), bind to seven sites on albumin. The combination of high plasma concentration, several binding sites, and a long half-life gives albumin an extensive capacity to transport fatty acids in the systemic compartments. Applicant conjugates CST74867-5418-2384.2Aty. Dkt. No.: 114198-3450 with 16-carbon fatty acid with a glutamic acid spacer to increase CST’s plasma half-life. In addition, Applicant modifies CST by acylating lysine at position 4 with a spacer and a CIS- fatty di-acid chain to further increase plasma half-life.

[0040] PEGylation of CST. When one or more chains of polyethylene glycol (PEG) is attached to a peptide, it changes the physical and chemical properties of peptides, such as its confrontation, electrostatic binding, and hydrophobicity, and results in an improvement in its pharmacokinetic profile. It enhances its half-life, peptide immunogenicity, and in-vivo stability. When PEG is attached to a therapeutic peptide, it protects the peptide from the immune system of the host, resulting in reduced immunogenicity. PEGylation of peptides decreases the rate of plasma clearance by preventing enzymatic degradation and prolongs their blood retention time. PEGylation of CST would increase the effective molecular weight of CST that is expected to shield CST from digestion by proteolytic enzymes via increased steric hindrance. CST can be easily PEGylated on a Lys residue.

[0041] A composition as disclosed herein can be a pharmaceutical composition. A “pharmaceutical composition” is intended to include the combination of an active agent with a carrier, inert or active, making the composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

[0042] “Pharmaceutically acceptable carriers” refers to any diluents, excipients, or carriers that may be used in the compositions disclosed herein. Pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field. They may be selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.

[0043] The compositions used in accordance with the disclosure can be packaged in dosage unit form for ease of administration and uniformity of dosage. The term "unit dose" or84867-5418-2384.2Aty. Dkt. No.: 114198-3450"dosage" refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the composition calculated to produce the desired responses in association with its administration, z.e., the appropriate route and regimen. The quantity to be administered, both according to number of treatments and unit dose, depends on the result and / or protection desired. Precise amounts of the composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the subject, route of administration, intended goal of treatment (alleviation of symptoms versus cure), and potency, stability, and toxicity of the particular composition. Upon formulation, solutions are administered in a manner compatible with the dosage formulation and in such amount as is therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described herein.

[0044] A combination as used herein intends that the individual active ingredients of the compositions are separately formulated for use in combination and can be separately packaged with or without specific dosages. The active ingredients of the combination can be administered concurrently or sequentially.

[0045] An “effective amount” is an amount sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications, or dosages. Such delivery is dependent on a number of variables including the time period for which the individual dosage unit is to be used, the bioavailability of the therapeutic agent, the route of administration, etc. It is understood, however, that specific dose levels of the therapeutic agents disclosed herein for any particular subject depends upon a variety of factors including the activity of the specific agent employed, bioavailability of the agent, the route of administration, the age of the animal and its body weight, general health, sex, the diet of the animal, the time of administration, the rate of excretion, the drug combination, and the severity of the particular disorder being treated and form of administration. In general, one will desire to administer an amount of the agent that is effective to achieve a serum level commensurate with the concentrations found to be effective in vivo. These considerations, as well as effective formulations and administration procedures are well known in the art and are described in standard textbooks.

[0046] “Therapeutically effective amount” of an agent refers to an amount of the agent that is an amount sufficient to obtain a pharmacological response; or alternatively, is an amount of the agent that, when administered to a patient with a specified disorder or disease, is94867-5418-2384.2Aty. Dkt. No.: 114198-3450 sufficient to have the intended effect, e.g., treatment, alleviation, amelioration, palliation or elimination of one or more manifestations of the specified disorder or disease in the patient. A therapeutic effect does not necessarily occur by administration of one dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations.

[0047] As used herein, the phrase “derived from” means isolated from, purified from, or engineered from, or any combination thereof.

[0048] In some embodiments, the terms “first” “second” “third” “fourth” or similar in a component name are used to distinguish and identify more than one component sharing certain identity in their names.

[0049] As used herein, “treating” or “treatment” of a disease in a subject refers to (1) preventing the symptoms or disease from occurring in a subject that is predisposed or does not yet display symptoms of the disease; (2) inhibiting the disease or arresting its development; or (3) ameliorating or causing regression of the disease or the symptoms of the disease. As understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of the present technology, beneficial or desired results can include one or more, but are not limited to, alleviation or amelioration of one or more symptoms, diminishment of extent of a condition (including a disease), stabilized (z.e., not worsening) state of a condition (including disease), delay or slowing of condition (including disease), progression, amelioration or palliation of the condition (including disease), states and remission (whether partial or total), whether detectable or undetectable. In one aspect, treatment excludes prophylaxis.

[0050] As used herein, the term “animal” refers to living multi-cellular vertebrate organisms, a category that includes, for example, mammals and birds. The term “mammal” includes both human and non-human mammals.

[0051] The term “subject,” “host,” “individual,” and “patient” are as used interchangeably herein to refer to animals, typically mammalian animals. Any suitable mammal can be treated by a method described herein. Non-limiting examples of mammals include humans, non- human primates (e.g., apes, gibbons, chimpanzees, orangutans, monkeys, macaques, and the like), domestic animals (e.g., dogs and cats), farm animals (e.g., horses, cows, goats, sheep, pigs) and experimental animals (e.g., mouse, rat, rabbit, guinea pig). In some embodiments, a mammal is a human. A mammal can be any age or at any stage of development (e.g., an104867-5418-2384.2Aty. Dkt. No.: 114198-3450 adult, teen, child, infant, or a mammal in utero). A mammal can be male or female. In some embodiments, a subject is a human. In some embodiments, a subject has or is diagnosed of having or is suspected of having a neurodegenerative disorder, e.g., a tauopathy.

[0052] Chromogranin A (protein: CgA; gene: CHGA in humans & Chga in mice) is a 48-kDa acidic secretory proprotein that gives rise to several peptides of biological importance, which include the dysglycemic hormone pancreastatin (PST: human CgA25o-3oi - amino acids 250- 301 of CHGA sequence as shown in SEQ ID NO: 29), the vasodilator vasostatin (human CgAi-76, i.e., amino acids 1-76 of SEQ ID NO: 29), and the anti-hypertensive peptide catestatin (CST: human CgA352-372, i.e., amino acids 352-372 of SEQ ID NO: 29) that inhibits nicotine-induced catecholamine release. Initially identified as a physiological brake in catecholamine secretion, CST has been established as a pleiotropic hormone having effects on promoting angiogenesis, lowering of blood pressure and cardiac contractility as well as enhancing baroreflex sensitivity and heart rate variability.

[0053] In addition to the above cardiovascular functions, CST has an antimicrobial activity and also regulates mast cell migration, cytokine production and release, smooth muscle cell proliferation, and monocyte migration. CST can act both extracellularly and intracellularly because the peptide can cross cell membrane.

[0054] In one embodiment, the term “disease” or “disorder” as used herein refers to a neurodegenerative disorder, e.g., a tauopathy, a status of being diagnosed with a neurodegenerative disorder, e.g., a tauopathy, a status of being suspect of having a neurodegenerative disorder, e.g., a tauopathy, or a status of at high risk of having a neurodegenerative disorder, e.g., a tauopathy.

[0055] A “neurodegenerative disorder” refers to a disorder showing a progressive loss of neurons, neuron structure, and / or their functions. Non-limiting examples of neurodegenerative disorders include Alzheimer’s disease, Parkinson’s disease, prion disease, Amyotrophic lateral sclerosis, motor neuron disease, Huntington’s disease, spinal muscular atrophy, and spinocerebellar ataxia.

[0056] As used herein, the term “cognitive decline” refers to a gradual decrease in a person's mental processes of perception, learning, memory, judgment, and reasoning. A “mild cognitive decline” refers to a decrease in a person's mental processes of perception, memory, judgment, and reasoning that is less than a 40% decrease, less than a 30% decrease, less than114867-5418-2384.2Aty. Dkt. No.: 114198-3450 a 20% decrease, or less than a 10% decrease as compared to the person's cognitive ability before the cognitive decline occurred.

[0057] As used herein the term “catestatin” or “CST” refers to a 21 amino acid long product of precursor hormone CgA. Wild type Human CST sequence is as shown as SEQ ID NO: 1. CgA is encoded by the Chromogranin A gene (GeneCards: CHGA, HGNC: 1929 NCBI Gene: 1113 Ensembl: ENSG00000100604 OMIM®: 118910 UniProtKB / Swiss-Prot: P10645).

[0058] As used herein, an “equivalent” when it refers to a polypeptide, is a polypeptide having at least one of the activities of a polypeptide in question. As used herein, a “CST equivalent” refers to a polypeptide that reduces Tau phosphorylation (e.g., S202 / T205 and / or S396 / S404 phosphorylation) or Tau aggregation by at least 50%, 60%, 70%, 80%, 90%, 95% or more in neuronal cells from a tauopathy model (e.g., PS19 mice). In some embodiments, an “equivalent of catestatin / CST” refers to a polypeptide selected from SEQ ID NOs: 1-28 or a retro, an inverse, or a retro-inverso of any one of the polypeptides as shown by SEQ ID NOs: 1-28.

[0059] “Administration” or “delivery” of CST or an equivalent thereof or a composition containing same can be performed in one dose, continuously or intermittently throughout the course of treatment. Methods of determining the most effective means and dosage of administration are known to those of skill in the art and will vary with the composition used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician or in the case of animals, by the treating veterinarian. Suitable dosage formulations and methods of administering the agents are known in the art. Route of administration can also be determined and method of determining the most effective route of administration are known to those of skill in the art and will vary with the composition used for treatment, the purpose of the treatment, the health condition or disease stage of the subject being treated, and target cell or tissue. Non-limiting examples of route of administration include oral administration, intraperitoneal, infusion, nasal administration, inhalation, injection, and topical application. In some embodiments, the administration is administration to a tumor microenvironment. In some embodiments, administering or a grammatical variation thereof also refers to more than one doses with certain interval. In some embodiments, the interval is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 10 days, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year124867-5418-2384.2Aty. Dkt. No.: 114198-3450 or longer. In some embodiments, one dose is repeated for once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times or more.

[0060] The term administration shall include without limitation, administration by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, intravascular, intraperitoneal, intracerebroventricular (ICV), intrathecal, intracistemal injection or infusion, intracranial, ocular, intradermally, percutaneously, subcutaneous injection, or implant), intratum orally, by inhalation spray nasal, intratracheal, vaginal, rectal, sublingual, urethral (e.g., urethral suppository) or topical routes of administration (e.g., gel, ointment, cream, aerosol, etc.) and can be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, excipients, and vehicles appropriate for each route of administration. The disclosure is not limited by the route of administration, the formulation or dosing schedule.

[0061] An agent of the present disclosure can be administered for therapy by any suitable route of administration. It will also be appreciated that the optimal route will vary with the condition and age of the recipient, and the disease being treated.

[0062] Administration or treatment in “combination” refers to administering two agents such that their pharmacological effects are manifest at the same time. Combination does not require administration at the same time or substantially the same time, although combination can include such administrations.

[0063] The phrase “first line” or “second line” or “third line” refers to the order of treatment received by a patient. First line therapy regimens are treatments given first, whereas second or third line therapy are given after the first line therapy or after the second line therapy, respectively.

[0064] As used herein, the term “purified” does not require absolute purity; rather, it is intended as a relative term. Thus, for example, a purified nucleic acid, peptide, protein, biological complexes, cell, virus or other active compound is one that is isolated in whole or in part from proteins or other contaminants. Generally, substantially purified peptides, proteins, biological complexes, cell, virus or other active compounds for use within the disclosure comprise more than 80% of all macromolecular species present in a preparation prior to admixture or formulation of the peptide, protein, biological complex, cell, virus or other active compound with a pharmaceutical carrier, excipient, buffer, absorption enhancing agent, stabilizer, preservative, adjuvant or other co-ingredient in a complete pharmaceutical134867-5418-2384.2Aty. Dkt. No.: 114198-3450 formulation for therapeutic administration. More typically, the peptide, protein, biological complex, cell, virus or other active compound is purified to represent greater than 90%, often greater than 95% of all macromolecular species present in a purified preparation prior to admixture with other formulation ingredients. In other cases, the purified preparation may be essentially homogeneous, wherein other macromolecular species are not detectable by conventional techniques.

[0065] In some embodiments, the term “engineered” or “recombinant” refers to having at least one modification not normally found in a naturally occurring protein, polypeptide, polynucleotide, strain, wild-type strain or the parental host strain of the referenced species. In some embodiments, the term “engineered” or “recombinant” refers to being synthetized by human intervention.

[0066] The terms “equivalent” or “biological equivalent” are used interchangeably when referring to a particular molecule, biological, or cellular material and intend those having minimal homology while still maintaining desired structure or functionality. Non-limiting examples of equivalent polypeptides, include a polypeptide having at least 60%, or alternatively at least 65%, or alternatively at least 70%, or alternatively at least 75%, or alternatively 80%, or alternatively at least 85%, or alternatively at least 90%, or alternatively at least 95% identity thereto or for polypeptide sequences, or a polypeptide which is encoded by a polynucleotide or its complement that hybridizes under conditions of high stringency to a polynucleotide encoding such polypeptide sequences. Conditions of high stringency are described herein and incorporated herein by reference. Alternatively, an equivalent thereof is a polypeptide encoded by a polynucleotide or a complement thereto, having at least 70%, or alternatively at least 75%, or alternatively 80%, or alternatively at least 85%, or alternatively at least 90%, or alternatively at least 95% identity, or at least 97% sequence identity to the reference polynucleotide, e.g., the wild-type polynucleotide.

[0067] Non-limiting examples of equivalent polynucleotides having at least 60%, or alternatively at least 65%, or alternatively at least 70%, or alternatively at least 75%, or alternatively 80%, or alternatively at least 85%, or alternatively at least 90%, or alternatively at least 95%, or alternatively at least 97%, identity to a reference polynucleotide. An equivalent also intends a polynucleotide or its complement that hybridizes under conditions of high stringency to a reference polynucleotide.144867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0068] An “equivalent” of a polynucleotide or polynucleotide region (or a polypeptide or polypeptide region) or a polyeptide or protein also includes a polynucleotide or polypeptide having a certain percentage (for example, 80%, 85%, 90%, 95%, or 98%) of “sequence identity” to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences. The alignment and the percent homology or sequence identity can be determined using software programs known in the art, for example those described in Current Protocols in Molecular Biology (Ausubel et al., eds. 1987) Supplement 30, section 7.7.18, Table 7.7.1. In certain embodiments, default parameters are used for alignment. A non-limiting exemplary alignment program is BLAST, using default parameters. In particular, exemplary programs include BLASTN and BLASTP, using the following default parameters: Genetic code=standard; filter=none; strand=both; cutoff=60; expect=10; Matrix=BLOSUM62; Descriptions=50 sequences; sort by=HIGH SCORE; Databases=non-redundant, GenBank+EMBL+DDBJ+PDB+GenBank CDS translations+SwissProtein+SPupdate+PIR. Details of these programs can be found at the following Internet address: ncbi.nlm.nih.gov / cgi-bin / BLAST. Sequence identity and percent identity can determined by incorporating them into clustalW (available at the web address:genome.jp / tools / clustalw / , last accessed on Jan. 13, 2017).

[0069] “Homology” or “identity” or “similarity” refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence that may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous at that position. A degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. An “unrelated” or “non-homologous” sequence shares less than 40% identity, or alternatively less than 25% identity, with one of the sequences of the present disclosure. In some embodiments, BLAST (accessible at blast.ncbi.nlm.nih.gov / Blast.cgi) or Clustal Omega (accessible at www.ebi.ac.uk / Tools / msa / clustalo / ) are used in determining the identity. In further embodiments, default setting is applied.

[0070] “Homology” or “identity” or “similarity” can also refer to two nucleic acid molecules that hybridize under stringent conditions.

[0071] “Hybridization” refers to a reaction in which one or more polynucleotides react to form a complex that is stabilized via hydrogen bonding between the bases of the nucleotide residues. The hydrogen bonding may occur by Watson-Crick base pairing, Hoogstein154867-5418-2384.2Aty. Dkt. No.: 114198-3450 binding, or in any other sequence-specific manner. The complex may comprise two strands forming a duplex structure, three or more strands forming a multi -stranded complex, a single self-hybridizing strand, or any combination of these. A hybridization reaction may constitute a step in a more extensive process, such as the initiation of a PCR reaction, or the enzymatic cleavage of a polynucleotide by a ribozyme.

[0072] Examples of stringent hybridization conditions include: incubation temperatures of about 25° C to about 37° C; hybridization buffer concentrations of about 6 / SSC to about 1 Ox SSC; formamide concentrations of about 0% to about 25%; and wash solutions from about 4*SSC to about 8*SSC. Examples of moderate hybridization conditions include: incubation temperatures of about 40° C to about 50° C; buffer concentrations of about 9 / SSC to about 2*SSC; formamide concentrations of about 30% to about 50%; and wash solutions of about 5 xSSC to about 2*SSC. Examples of high stringency conditions include: incubation temperatures of about 55° C to about 68° C; buffer concentrations of about 1 xSSC to about O. l xSSC; formamide concentrations of about 55% to about 75%; and wash solutions of about 1 xSSC, 0.1 xSSC, or deionized water. In general, hybridization incubation times are from 5 minutes to 24 hours, with 1, 2, or more washing steps, and wash incubation times are about 1, 2, or 15 minutes. SSC is 0.15 M NaCl and 15 mM citrate buffer. It is understood that equivalents of SSC using other buffer systems can be employed.

[0073] “Eukaryotic cells” comprise all of the life kingdoms except monera. They can be easily distinguished through a membrane-bound nucleus. Animals, plants, fungi, and protists are eukaryotes or organisms whose cells are organized into complex structures by internal membranes and a cytoskeleton. The most characteristic membrane-bound structure is the nucleus. Unless specifically recited, the term “host” includes a eukaryotic host, including, for example, yeast, higher plant, insect and mammalian cells. Non-limiting examples of eukaryotic cells or hosts include simian, bovine, porcine, murine, rat, avian, reptilian and human.

[0074] “Prokaryotic cells” that usually lack a nucleus or any other membrane-bound organelles and are divided into two domains, bacteria and archaea. In addition to chromosomal DNA, these cells can also contain genetic information in a circular loop called on episome. Bacterial cells are very small, roughly the size of an animal mitochondrion (about 1-2 pm in diameter and 10 pm long). Prokaryotic cells feature three major shapes: rod shaped, spherical, and spiral. Instead of going through elaborate replication processes like164867-5418-2384.2Aty. Dkt. No.: 114198-3450 eukaryotes, bacterial cells divide by binary fission. Examples include but are not limited to Bacillus bacteria, E. coli bacterium, and Salmonella bacterium.

[0075] As used herein, a “retro” version of CST or a CST equivalent refers to a reversal of sequence (carboxyl— >amino). As an example, the retro version of the tripeptide “SMK” would be “KMS.”

[0076] As used herein, an “inverso” version of CST or a CST equivalent refers to an inversion of chirality (L— D amino acids). Glycine is an achiral amino acid, so its L and D orientations are the same. The inverso version of the tripeptide “SMK” would be “smk” where lower case letters represent D amino acids.

[0077] As used herein, a “retro-inverso” or “(R-I)” version of CST or a CST equivalent refers to an inversion of chirality (L— D amino acids) and also reversal of sequence(carboxyl— >amino). Glycine is an achiral amino acid, so its L and D orientations are the same. R-I peptide approach provides a general method for generation of metabolically stable mimics of biologically active peptides for diagnostics and therapeutics. The retro-inverso version of the tripeptide “SMK” would be “kms” where lower case letters represent D amino acids.

[0078] As used herein, “Tau” refers to a human protein encoded by the microtubule- associated protein Tau gene, MAPT, located on chromosome 17q21. Although Tau is usually highly soluble, under pathological conditions, it can aggregate into paired helical filaments, neurofibrillary tangles and other structures that define a large spectrum of neurodegenerative diseases termed tauopathies. Phosphorylation is one of the most prevalent post-translational modifications found in aggregated tau in tauopathies.

[0079] As used herein, a “tauopathy” refers to a class of neurodegenerative diseases associated with aggregation of the microtubule-associated protein Tau, including AD, PSP, Down's syndrome, frontotemporal lobe dementia, frontotemporal dementia, Pick's disease, CBD, or AGD.Modes for Carrying Out the DisclosureA non-naturally modified CST or an equivalent thereof and compositions containing same174867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0080] In one aspect, the disclosure provides non-naturally modified CST peptides or equivalents thereof and compositions containing same. The modified CST peptides have longer half-life and / or better bioavailability.

[0081] In some embodiments, the CST or the equivalent thereof further comprises a nonnatural amino acid at position R8 of SEQ ID NO: 1, or an equivalent thereof in a CST sequence, e.g., a CST sequence selected from SEQ ID NOs: 2-28. In some embodiments, the non-natural amino acid is D-Arginine, a methyl amino acid (e.g., a methyl arginine) or a P- amino acid (e.g., a beta arginine).

[0082] In some embodiments, the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons. In some embodiments, the fatty acid is fully saturated, monounsaturated or polyunsaturated. In some embodiments, the fatty acid is attached to the CST or the equivalent thereof via a linker.

[0083] In some embodiments, the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety. In some embodiments, the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety. In some embodiments, the at least one PEG moiety is attached at a Lysine residue on the CST or equivalent thereof. In some embodiments, the CST or the equivalent thereof further comprises at least two, at least three, at least four or at least five PEG moieties.

[0084] Another aspect of the disclosure is directed to a composition comprising a non- naturally modified CST or an equivalent thereof, wherein the non-natural modification one or more modifications selected from (a) a non-natural amino acid; (b) a fatty acid or (c) polyethylene glycol (PEG).

[0085] In some embodiments, the non-natural amino acid is at position R8.

[0086] In some embodiments, the non-natural amino acid is a D-Arginine, a methyl amino acid or a P-amino acid.

[0087] In some embodiments, the fatty acid has 10, 12, 14, 16, or 18 carbons. In a specific embodiment, the fatty acid has 16 carbons.

[0088] In some embodiments, the fatty acid is fully saturated, monounsaturated or polyunsaturated. In some embodiments, the fatty acid is atached to the CST or the equivalent thereof via a linker.184867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0089] In some embodiments, the fatty acid is attached to the CST or the equivalent thereof via a linker.

[0090] In some embodiments, the CST or the equivalent thereof comprises at least one PEG moiety.

[0091] In some embodiments, the CST or the equivalent thereof comprises between two and five PEG moieties.

[0092] In some embodiments, the PEG moiety is attached at a lysine residue of the CST or the equivalent thereof.

[0093] In some embodiments, the compositions of the present disclosure further comprise a pharmaceutically acceptable carrier.Methods of Treatment

[0094] This disclosure is based, at least in part, of the surprising neuroprotective effects of CST or an equivalent thereof in neurodegenerative diseases, especially for the treatment and prevention of tauopathies.

[0095] In one aspect, provided herein is a method for treating neurodegeneration in a subject in need thereof comprising, or alternatively consisting of CST or an equivalent thereof to the subject.

[0096] In some embodiments, the effective amount of CST is an amount that results in at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% decrease in phosphorylation of Tau in the subject. In some embodiments, Tau phosphorylation is measured from the plasma of the subject. Tau phosphorylation can be detected and measured from plasma samples using biomarker detection assays, such as phospho-Tau (Catalog #: K15121D-1) and total Tau (Catalog #: K151APSS-1) assays from Meso Scale Discovery™.

[0097] In some embodiments, the effective amount of CST is an amount that results in an improvement in memory function of the subject. In some embodiments, the memory function of the subject is measured by the Montreal Cognitive Assessment (MoCA), which involves memorizing a short list of words, copying a drawing, and identifying pictures of animals. In some embodiments, the memory function of the subject is measured by the MiniMental State Exam (MMSE), which includes tasks like counting backwards, naming objects,194867-5418-2384.2Aty. Dkt. No.: 114198-3450 and following simple instructions. In some embodiments, the memory function of the subject is measured by the Mini-Cog test, which combines a three-item recall test with a clockdrawing task. In some embodiments, the memory function of the subject is measured by the Memory Impairment Screen (MIS) assesses memory through word recall tasks.

[0098] In some embodiments, the effective amount of CST is between about 20mg CST to about 200mg CST (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, or 200 mg).

[0099] In some embodiments, the effective amount of CST is between about 0.2 mg / kg to about 5 mg / kg (about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.3, 1.5, 1.7, 2, 2.3, 2.7, 3, 3.3, 3.7, 4, 4.3, 4.7, or 5 mg / kg).

[0100] In some embodiments, the subject has low levels of naturally occurring CST in the subject’s plasma, cortex or hippocampus.

[0101] In some embodiments, the subject is suffering from a tauopathy. In some embodiments, the tauopathy is selected from AD, PSP, Down's syndrome, frontotemporal lobe dementia, frontotemporal dementia, Pick's disease, CBD, or AGD

[0102] In a specific embodiment, the tauopathy is AD. In some embodiments, AD in the subject is at least stage 4 based on the Braak scale (e.g., Braak 4, Braak 5, or Braak 6).

[0103] In some embodiments, wherein the CST or the equivalent thereof comprises, or alternatively consists essentially of, or consists of, or yet further consists of a peptide selected from SEQ ID NOs: 1-28.

[0104] In some embodiments, the CST or the equivalent thereof further comprises a nonnatural amino acid at position R8 of SEQ ID NO: 1, or an equivalent thereof in a CST sequence, e.g., a CST sequence selected from SEQ ID NOs: 2-28. In some embodiments, the non-natural amino acid is D-Arginine, a methyl amino acid (e.g., a methyl arginine) or a P- amino acid (e.g., a beta arginine).

[0105] In some embodiments, the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons. In some embodiments, the fatty acid is fully saturated, monounsaturated or polyunsaturated. In some embodiments, the fatty acid is attached to the CST or the equivalent thereof via a linker.204867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0106] In some embodiments, the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety. In some embodiments, the at least one PEG moiety is attached at a Lysine residue on the CST or equivalent thereof. In some embodiments, the CST or the equivalent thereof further comprises at least two, at least three, at least four or at least five PEG moieties.

[0107] In some embodiments, the equivalent of CST comprises, or alternatively consists essentially of, or consists of, or yet further consists of a retro-inverso peptide of any of the sequences selected from SEQ ID NOs: 1-28.

[0108] In some embodiments, the CST or an equivalent thereof is administered every 12 hours, once every day, once every three days, once every week, once in two weeks or once every month.

[0109] In some embodiments, the subject is a mammal, optionally a human patient.

[0110] In some embodiments, the CST or an equivalent thereof is administered by an enteral route, buccal route, intraperitoneal route, inhalation route, intravenous route, subcutaneous route or intramuscular route.[OHl] The following examples are included to demonstrate some embodiments of the disclosure. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.Experimental MethodsExperimental Method No. 1 - CST Levels are Decreased in AD Patients and AD mouse models.

[0112] Catestatin levels were measured in samples from the cortex, hippocampus or cerebrospinal fluid (CSF) of AD patients having mild disease (Braak 1-2) or severe disease (Braak 6). It was observed that CSF concentration in brain decreases as neurodegeneration progresses (in AD, from Braak 1-2 to Braak 6) (See, FIGS. 1A&1B). The observation that CST concentration is reduced neurodegenerative diseases was also confirmed in an AD mouse model (PS 19) (FIG. 2B&C).Experimental Method No. 2 - CST Treatment decreases Tau phosphorylation in vivo.214867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0113] Applicant used organotypic slice culture (OTSC) taken from the hippocampus of WT pups (7-8 day old) to examine the direct role of CST in tau seeding by the repeat domains of Tau (KI 8) that comprises the core of Tau fibrils. Western blotting with phospho-Tau antibodies and immunostaining of slices with Tau oligomer-specific MCI antibody revealed that CST treatment attenuated phospho-Tau and caused a -59% decrease in Tau aggregates (FIGS. 3B-3C), implicating a role for CST in ameliorating Tau proteinopathy.

[0114] To determine whether CST ameliorates tauopathy, Applicant treated 5-month-old PS19 mice with intraperitoneal CST (0.5 pg / g BW; 3 times / week) until they were 9 months old (FIG. 4A). Applicant found that CST caused a -73% decrease in phosphorylation of Tau at S202 / T205 and S396 / S404 (FIGS. 4B-4C) in this tauopathy disease model, indicating CST regulation of Tau proteinopathy.

[0115] To determine whether CST decreases Tau aggregation, Applicant treated 5-month-old PS19 with intraperitoneal CST (0.5 pg / g BW; 3 times / week) until they were 9 months old (FIG. 4A). Applicant found a -76% decrease in Tau aggregation in both CA3 (5B) and dentate gyrus (5D) regions.Embodiments

[0116] This disclosure further includes the following non-limiting embodiments:

[0117] Embodiment 1. A method for treating neurodegeneration in a subject in need thereof comprising administering an effective amount of CST or an equivalent thereof to the subject.

[0118] Embodiment 2. The method of embodiment 1, wherein the effective amount of CST is an amount that results in (i) at least 50% decrease in phosphorylation of Tau in the subject or (ii) an improvement in memory function of the subject.

[0119] Embodiment 3. The method of embodiment 1 or embodiment 2, wherein the effective amount is between about 20 mg to about 200 mg.

[0120] Embodiment 4. The method of any one of embodiments 1-3, wherein the subject has low levels of naturally occurring CST in the subject’s plasma cortex or hippocampus.

[0121] Embodiment 5. The method of any one of embodiments 1-3, wherein the subject is suffering from a tauopathy.

[0122] Embodiment 6. The method of any one of embodiments 1-5, wherein the CST or the equivalent thereof comprises a peptide selected from SEQ ID NOs: 1-28.224867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0123] Embodiment 7. The method of embodiment 6, wherein the CST or the equivalent thereof further comprises a non-natural amino acid at position R8 of SEQ ID NO: 1, or an equivalent thereof in a CST sequence wherein the non-naturally amino acid at position R8 is retained or a CST sequence selected from SEQ ID NOs: 2-28.

[0124] Embodiment 8. The method of embodiment 6, wherein the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons.

[0125] Embodiment 9. The method of embodiment 6, wherein the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety.

[0126] Embodiment 10. The method of any one of embodiments 1-5, wherein the equivalent of CST comprises a retro-inverso peptide of any of the sequences selected from SEQ ID NOs: 1-28.

[0127] Embodiment 11. The method of any one of embodiments 1-10, wherein the CST or an equivalent thereof is administered every 12 hours, once every day, once every three days, once every week, once in two weeks or once every month.

[0128] Embodiment 12. The method of any one of embodiments 1-11, wherein the subject is a mammal, optionally a human patient.

[0129] Embodiment 13. The method of any one of embodiments 1-12, wherein the CST or an equivalent thereof is administered by an enteral route, buccal route, intraperitoneal route, inhalation route, intravenous route, subcutaneous route or intramuscular route.

[0130] Embodiment 14. A CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises a nonnatural amino acid at position R8, and compositions comprising the CST or the equivalent thereof.

[0131] Embodiment 15. The CST of embodiment 14, wherein the non-natural amino acid is a D-Arginine, a methyl amino acid or a P-amino acid.

[0132] Embodiment 16. A CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons and compositions comprising the CST or the equivalent thereof.234867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0133] Embodiment 17. The CST or composition of embodiment 16, wherein the fatty acid is attached to the CST or the equivalent thereof comprises 16 carbons.

[0134] Embodiment 18. A CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety and compositions comprising the CST or the equivalent thereof.

[0135] Embodiment 19. The CST or composition of embodiment 18, wherein the PEG moiety is atached at a Lysine residue of the CST or the equivalent thereof.

[0136] Embodiment 20. A non-naturally modified CST or an equivalent thereof, wherein the non-natural modification one or more modifications selected from (a) a non-natural amino acid; (b) a fatty acid or (c) polyethylene glycol (PEG) and compositions comprising the CST or the equivalent thereof.

[0137] Embodiment 21. The CST or composition of embodiment 20, wherein the non-natural amino acid is at position R8.

[0138] Embodiment 22. The CST or composition of embodiment 20 or embodiments 21, wherein the non-natural amino acid is a D-Arginine, a methyl amino acid or a P-amino acid.

[0139] Embodiment 23. The CST or composition of embodiment 20, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons.

[0140] Embodiment 24. The CST or composition of embodiment 20 or embodiment 23, wherein the fatty acid is attached to the CST or the equivalent thereof via a linker.

[0141] Embodiment 25. The CST or composition of embodiment 20, wherein the CST or the equivalent thereof comprises at least one PEG moiety.

[0142] Embodiment 26. The CST or composition of embodiment 20 or embodiment 25, wherein the CST or the equivalent thereof comprises between two and five PEG moieties.

[0143] Embodiment 27. The CST or composition of any one of embodiments 20, 25 or 26, wherein the PEG moiety is attached at a lysine residue of the CST or the equivalent thereof.244867-5418-2384.2Aty. Dkt. No.: 114198-3450Sequence Listing

[0144] SEQ ID NO: 1, human (Homo sapiens) CST sequence CHGA352-372,SSMKLSFRARAYGFRGPGPQL

[0145] SEQ ID NO: 2, Human Gly364Ser variant sequence CHGA352-372,S SMKLSFRARAYSFRGPGPQL

[0146] SEQ ID NO: 3, Human Gly364Val variant sequence CHGA352-372,SSMKLSFRARAYGFRVPGPQL

[0147] SEQ ID NO: 4, Human Pro370Leu variant sequence CHGA352-372,SSMKLSFRARAYGFRGPGLQL

[0148] SEQ ID NO: 5, Human CST-R variant sequence CHGA352-373,SSMKLSFRARAYGFRGPGLQLR

[0149] SEQ ID NO: 6, Sumatran orangutan (Pongo abelli) CST sequence CHGA352-372,S SMKLPFRARAYGFRGPGPQL

[0150] SEQ ID NO: 7, Black snub-nosed monkey (Rhinopithecus bieti) CST sequence CHGA348-368,RSMKLSFRARAYGFRGPGPQL

[0151] SEQ ID NO: 8, Night monkey (Aotus nancymaae) CST sequence CHGA351-371,HSMKLSFQARAYGFRGPGPQL

[0152] SEQ ID NO: 9, Sooty mangabey (Cercocebus atys) CST sequence CHGA354-374,GSMKLSFRARAYGFRGPGPQL

[0153] SEQ ID NO: 10, Tufted capuchin (Sapajus apella) CST sequence CHGA351-371,HSMKLSFRGRAYGFRGPGPQL

[0154] SEQ ID NO: 11, brown rat (Rattus norvegicus) CST sequence CHGA366-386,RSMKLSFRARAYGFRDPGPQL

[0155] SEQ ID NO: 12, Golden hamster (Mesocricetus auratus) CST sequence CHGA335- 355,RSMKLSFRARAYGFRDPGPQP254867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0156] SEQ ID NO: 13, Grasshopper mouse (Onychomys torridus) CST sequence CHGA347-367,RSMKLPFRARAYGFRDPGPQL

[0157] SEQ ID NO: 14, House mouse (Mus musculus) CST sequence CHGA364-384,RSMKLSFRTRAYGFRDPGPQL

[0158] SEQ ID NO: SEQ ID NO: SEQ ID NO: 15, European rabbit (Oryctolagus cuniculus) CST sequence CHGA342-362,RSMKLSFQPRAYGFRGPGPQL

[0159] SEQ ID NO: 16, Arabian camel (Camelus dromedarius) CST sequence CHGA347- 367,HSMKLSFRARAYGFRGPGLQL

[0160] SEQ ID NO: 17, Cattle (Bos taurus) CST sequence CHGA344-364,RSMRLSFRARGYGFRGPGLQL

[0161] SEQ ID NO: 18, Goat (Capra hircus) CST sequence CHGA338-358,RSMRLSFQARGYGFRGPGLQL

[0162] SEQ ID NO: 19, Pig (Sus scrofa) CST sequence CHGA343-363,RSMKLSFRAPAYGFRGPGLQL

[0163] SEQ ID NO: 20, Horse (Equus caballus) CST sequence CHGA343-363,RSMKLSFRARAYGFRGPGLQL

[0164] SEQ ID NO: 21, California sea lion (Zalophus californianus) CST sequence CHGA344-364,RSMRLSFRARAYDFRGPGLQL

[0165] SEQ ID NO: 22, Cat (Fells ca s') CST sequence CHGA349-369,SSMKLSFRARAYDFRGPGLQL

[0166] SEQ ID NO: 23, Dog (Cams lupus) CST sequence CHGA343-363,HSMKLSFRARAYDFRGPGLPL264867-5418-2384.2Aty. Dkt. No.: 114198-3450

[0167] SEQ ID NO: 24, Giant panda (Ailuropoda melanoleucd) CST sequence CHGA346-366,RSMKLSFRARAYDFRGPGLQL

[0168] SEQ ID NO: 25, Mongooses (Suricata suricatta) CST sequence CHGA350-370,RSMKLAFRARAYDFRGPGLQL

[0169] SEQ ID NO: 26, Dolphin (Lipotes vexillifer) CST sequence CHGA339-359,RAMKLSFRARGYDF SGPGLPL

[0170] SEQ ID NO: 27, Killer whale (Orcinus orca) CST sequence CHGA340-360,RAMKLSFRARGYDF SGPGLQL

[0171] SEQ ID NO: 28, Sperm whale (Physeter catodon) CST sequence CHGA355-375,RAMKLSFRARGYDFRGPGLQL

[0172] SEQ ID NO: 29: Homo sapiens CHGA full length sequenceLPVNSPMNKGDTEVMKCIVEVISDTLSKPSPMPVSQECFETLRGDERILSILRHQNLL KELQDLALQGAKERAHQQKKHSGFEDELSEVLENQSSQAELKEAVEEPSSKDVMEK REDSKEAEKSGEATDGARPQALPEPMQESKAEGNNQAPGEEEEEEEEATNTHPPASL PSQKYPGPQAEGDSEGLSQGLVDREKGLSAEPGWQAKREEEEEEEEEAEAGEEAVPE EEGPTVVLNPHPSLGYKEIRKGESRSEALAVDGAGKPGAEEAQDPEGKGEQEHSQQKEEEEEMAVVPQGLFRGGKSGELEQEEERLSKEWEDSKRWSKMDQLAKELTAEKR LEGQEEEEDNRDSSMKLSFRARAYGFRGPGPQLRRGWRPSSREDSLEAGLPLQVRG YPEEKKEEEGSANRRPEDQELESLSAIEAELEKVAHQLQALRRGEquivalents

[0173] 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 technology belongs.

[0174] The present technology illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention274867-5418-2384.2Aty. Dkt. No.: 114198-3450 in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the present technology claimed.

[0175] Thus, it should be understood that the materials, methods, and examples provided here are representative of preferred aspects, are exemplary, and are not intended as limitations on the scope of the present technology.

[0176] It should be understood that although the present invention has been specifically disclosed by certain aspects, embodiments, and optional features, modification, improvement and variation of such aspects, embodiments, and optional features can be resorted to by those skilled in the art, and that such modifications, improvements and variations are considered to be within the scope of this disclosure.

[0177] The present technology has been described broadly and generically herein. Each of the narrower species and sub-generic groupings falling within the generic disclosure also form part of the present technology. This includes the generic description of the present technology with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0178] In addition, where features or aspects of the present technology are described in terms of Markush groups, those skilled in the art will recognize that the present technology is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0179] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference individually. In case of conflict, the present specification, including definitions, will control.

[0180] Other aspects are set forth within the following claims.284867-5418-2384.2

Claims

1. Aty. Dkt. No.: 114198-3450WHAT IS CLAIMED IS:

1. A method for treating neurodegeneration in a subject in need thereof comprising administering an effective amount of catestatin (CST) or an equivalent thereof to the subject.

2. The method of claim 1, wherein the effective amount of CST is an amount that results in (i) at least 50% decrease in phosphorylation of Tau in the subject or (ii) an improvement in memory function of the subject.

3. The method of claim 1 or claim 2, wherein the effective amount is between about 20 mg to about 200 mg.

4. The method of any one of claims 1-3, wherein the subject has low levels of naturally occurring CST in the subject’s plasma cortex or hippocampus.

5. The method of any one of claims 1-3, wherein the subject is suffering from a tauopathy.

6. The method of any one of claims 1-5, wherein the CST or the equivalent thereof comprises a peptide selected from SEQ ID NOs: 1-28.

7. The method of claim 6, wherein the CST or the equivalent thereof further comprises a non-natural amino acid at position R8 of SEQ ID NO: 1, or an equivalent thereof.

8. The method of claim 6, wherein the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons.

9. The method of claim 6, wherein the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety.

10. The method of any one of claims 1-5, wherein the equivalent of CST comprises a retro-inverso peptide of any of the sequences selected from SEQ ID NOs: 1-28.

11. The method of any one of claims 1-10, wherein the CST or an equivalent thereof is administered every 12 hours, once every day, once every three days, once every week, once in two weeks or once every month.

12. The method of any one of claims 1-11, wherein the subject is a mammal, optionally a human patient.

13. The method of any one of claims 1-12, wherein the CST or an equivalent thereof is administered by an enteral route, buccal route, intraperitoneal route, inhalation route, intravenous route, subcutaneous route or intramuscular route.294867-5418-2384.2Aty. Dkt. No.: 114198-345014. A composition comprising a CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises a non-natural amino acid at position R8.

15. The composition of claim 14, wherein the non-natural amino acid is D-Arginine, a methyl amino acid or a P-amino acid.

16. A composition comprising a CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises a fatty acid and / or an acyl group, wherein the fatty acid has 10, 12, 14, 16, or 18 carbons.

17. The composition of claim 16, wherein the fatty acid is attached to the CST or the equivalent thereof comprises 16 carbons.

18. A composition comprising a CST or an equivalent thereof that comprises a peptide selected from SEQ ID NOs: 1-28, wherein the CST or the equivalent thereof further comprises at least one polyethylene glycol (PEG) moiety.304867-5418-2384.2

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