Novel composition and methods of treating a neurodegenerative disease

Abstract

Compositions and methods to inhibit the interaction between two protein molecules, PTEN and PSD-95, in neurons are provided. The inhibition of the interaction between PTEN and PSD-95 is used in methods of treating Alzheimer's disease. The compositions comprise isochlorogenic acid B, isochlorogenic acid C, and amaranth.

Description

[0001] DESCRIPTION

[0002] TITLE

[0003] NOVEL COMPOSITION AND METHODS OF TREATING A NEURODEGENERATIVE DISEASE

[0004] CROSS-REFERENCE TO RELATED APPLICATION

[0005] The present application claims the benefit of U.S. Provisional Application Serial No. 63 / 580,829, filed September 6, 2023, which is hereby incorporated by reference herein in its entirety.

[0006] SEQUENCE LISTING

[0007] The Sequence Listing for this application is labeled “CUHK-218XClPCT-SeqList- 05Sep24.xml” which was created on September 5, 2024 and is 2,242 bytes. The entire contents of the sequence listing is incorporated herein by reference in its entirety.

[0008] FIELD OF THE INVENTION

[0009] The invention relates to the use of novel pharmaceutical compositions and methods for their use in the treatment and prevention of a neurodegenerative disease, including Alzheimer’ s disease.

[0010] BACKGROUND OF THE INVENTION

[0011] Alzheimer's disease (AD) is the most common form of dementia affecting the aging population. Its pathology involves the accumulation of amyloid plaques and neurofibrillary tangles in the patient’s brain. Cognitive impairment, regarded as an early manifestation of AD, is believed to be attributable to disruptions of synaptic functions. At the present time, there are an estimated 55 million AD sufferers worldwide, and the number is expected to increase to at least 78 million by the year 2030 and over 139 million by the year 2050. AD is the 6thleading cause of death in the United States and ranked the 3rdleading cause of death among elderly people. Although AD symptoms typically begin to emerge in a patient's late 60s or 70s, early onset of AD can be seen among patients in their 40s or even late 30s. Currently there is no cure for AD, and AD patients experience a gradual deterioration over time until their death. The prevalence of AD and demands for AD patient treatment and maintenance have significant and profound socio-economic implications. In the US alone, the annual cost related to AD patient care is well over 600 billion dollars.

[0012] Accordingly, there exists a need for novel and effective methods and compositions for treating AD and other neurodegenerative disorders by alleviating the pertinent symptoms and / or delaying the onset of such symptoms.

[0013] BRIEF SUMMARY OF THE INVENTION

[0014] The subject invention pertains to novel compositions and methods to inhibit the interaction of phosphatase and tensin homologue deleted from chromosome 10 (PTEN) and PSD95 / Disc large / Zonula occludens-1 (PDZ). In certain embodiments, the subject methods can be used to treat neurodegenerative disorders. PTEN (phosphatase and tensin homologue deleted from chromosome 10) is a lipid phosphatase with specificity towards phosphatidylinositol-3,4,5-trisphosphate (PIP3) and a key negative regulator of the phosphatidylinositol-3 -kinase (PI3K). The activity of PI3K favors the formation of the PIP3 required for AMPA receptor (AMP AR) activity at synapses while turnover of PIP3 by PTEN facilitates AMP AR endocytosis during long-term depression (LTD). LTD is a form of synaptic plasticity characterized by a long-lasting reduction in synaptic activity and is extensively implicated in memory loss in AD.

[0015] In certain embodiments, the NMDAR-dependent LTD is mediated by interaction between PTEN and PSD95 / Disc large / Zonula occludens-1 (PDZ) proteins, which leads to the recruitment and anchoring of PTEN at the postsynaptic density in dendrite spines. The domain in PTEN responsible is the PDZ binding motif. Treating with Ap leads to recruitment of PTEN to the dendritic spines, which demonstrates a link between PTEN and AD.

[0016] In certain embodiments, compounds and compositions that directly modulate the interactions of PTEN with PDZ proteins can be used to inhibit PTEN:PDZ interactions.

[0017] In certain embodiments, at least 1, 2, 3, 4, or 5 molecules can be used to inhibit PTEN:PDZ interactions. In certain embodiments, the molecule is amaranth, isochlorogenic acid B, isochlorogenic C, pentagalloylglucose, hematoporphyrin dihydrochloride, or any combination thereof. In certain embodiments, the at least 1, 2, 3, 4, or 5 molecules can dock in the PSD-95 PDZ binding pocket and inhibit the PTEN:PDZ interactions.

[0018] In certain embodiments, the PTEN:PDZ interaction inhibitors, such as, for example, amaranth, isochlorogenic acid B, isochlorogenic C, pentagalloylglucose, hematoporphyrin dihydrochloride, or any combination thereof individually can result in improvement of patients suffering from a neurodegenerative disorder, such as, for example, Alzheimer's Disease (AD), including both prophylactically and therapeutically.

[0019] BRIEF DESCRIPTION OF THE SEQUENCES

[0020] SEQ ID NO: 1 : fluorescent-labeled peptide

[0021] BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIGs. 1A-1B show the binding curves for FITC-PTEN-PDZ-motif probe and PDZ proteins (PSD95 and MAGI2) interactions. Values are mean ± SD.

[0023] FIGs. 2A-2B show the inhibitory effects of 23 candidate compounds in disrupting interaction between PTEN and PSD95 / MAGI2. Values are mean ± SD.

[0024] FIGs. 3A-3K show the IC50 curves for the 11 compounds with the lowest IC50 values. The graphs showed the level of polarization (mP) against compound concentration. Values are mean ± SD.

[0025] FIGs. 4A-4B show the ability of promising compounds (200pM) in blocking binding between PTEN and PSD95 in GST pull down and co-immunoprecipitation assay.

[0026] FIGs. 5A-5B show the neurotoxicity effect of #58 amaranth, #80 pentagalloylglucose, #82 isochl orogenic acid C, #84 isochlorogenic acid B and #148 hematoporphyrin dihydrochloride in (FIG. 5A) iPSC-derived neural progenitor cells and (FIG. 5B) iPSC- derived neurons by MTS assay. Values are mean ± SD.

[0027] FIGs. 6A-6B. FIG. 6A shows the effect of Ap 1-42 (lOpM) in increasing the association between PSD95 and PTEN by co-immunoprecipitation of PSD95 and PTEN from iPSC-NPC lysates incubating with lOpM AP(i-42) for 24 hours. FIG. 6B shows the partially abolishment of PSD95:PTEN binding induced by AP1-42 in iPSC-neuron after #82 and #84 treatment.

[0028] FIG. 7 shows the inhibitory effects of #84 isochlorogenic acid B and #82 isochlorogenic acid C in disrupting interaction between PTEN and PSD95 in 7-8-month 5xFAD synaptosomes.

[0029] FIGs. 8A-8B show the neuroprotective effects of #82 isochlorogenic acid C (100 pM) and #84 isochlorogenic acid B (100 pM) against AP1-42 (10 pM) in iPSC-derived neurons.

[0030] FIGs. 9A-9H show the effect of an embodiment of #58 amaranth, #80 pentagalloylglucose, #82 isochlorogenic acid C, #84 isochlorogenic acid B and #148 hematoporphyrin dihydrochloride on rescuing the memory deficits of 5xFAD mice in Morris Water Maze test. All Data presented as mean ± SEM (#84 isochl orogenic acid B and #82 isochlorogenic acid C treatment, n=13; #148 hematoporphyrin dihydrochloride treatment, n=10; #58 amaranth treatment, n=10; #80 pentagalloylglucose treatment, n=7). Escape latency was quantified for (FIGs. 9A-9B) invisible platform (Day 0, one-way ANOVA), (FIGs. 9C- 9D) training trials (Day 1-4, two-way ANOVA, @ / ?<0.05, ### / ?<0.001 and compared with 5xFAD control group,+ / ?<0.01 and "' / KO.01 compared with WT control group). FIGs. 9E-9H showed the number of entries and escape latency for probe trial (Day 5, one-way ANOVA, K0.05, ** / K0.01 and *** / K0.001).

[0031] DETAILED DISCLOSURE OF THE INVENTION

[0032] Selected Definitions

[0033] As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent that the terms “including”, “includes”, “having”, “has”, “with”, or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising”. The transitional terms / phrases (and any grammatical variations thereof) “comprising”, “comprises”, “comprise”, “consisting essentially of’, “consists essentially of’, “consisting” and “consists” can be used interchangeably.

[0034] The phrases “consisting essentially of’ or “consists essentially of’ indicate that the claim encompasses embodiments containing the specified materials or steps and those that do not materially affect the basic and novel characteristic(s) of the claim.

[0035] The term “about” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured, z.e., the limitations of the measurement system. In the context of compositions containing amounts of ingredients where the terms “about” is used, these compositions contain the stated amount of the ingredient with a variation (error range) of 0-10% around the value (X ± 10%). In other contexts the term “about” is provides a variation (error range) of 0-10% around a given value (X ± 10%). As is apparent, this variation represents a range that is up to 10% above or below a given value, for example, X ± 1%, X ± 2%, X ± 3%, X ± 4%, X ± 5%, X ± 6%, X ± 7%, X ± 8%, X ± 9%, or X ± 10%.

[0036] In the present disclosure, ranges are stated in shorthand to avoid having to set out at length and describe each and every value within the range. Any appropriate value within the range can be selected, where appropriate, as the upper value, lower value, or the terminus of the range. For example, a range of 0.1 -1.0 represents the terminal values of 0.1 and 1.0, as well as the intermediate values of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and all intermediate ranges encompassed within 0.1-1.0, such as 0.2-0.5, 0.2-0.8, 0.7-1.0, etc. Values having at least two significant digits within a range are envisioned, for example, a range of 5-10 indicates all the values between 5.0 and 10.0 as well as between 5.00 and 10.00 including the terminal values. When ranges are used herein, combinations and subcombinations of ranges (e.g., subranges within the disclosed range) and specific embodiments therein are explicitly included.

[0037] As used herein, the terms “therapeutically-effective amount,” “therapeutically-effective dose,” “effective amount,” and “effective dose” are used to refer to an amount or dose of a compound or composition that, when administered to a subject, is capable of treating, preventing, or improving a condition, disease, or disorder in a subject. In other words, when administered to a subject, the amount is “therapeutically effective.” The actual amount will vary depending on a number of factors including, but not limited to, the particular condition, disease, or disorder being treated, prevented, or improved; the severity of the condition; the weight, height, age, and health of the patient; and the route of administration.

[0038] As used herein, the term “treatment” refers to eradicating; reducing; ameliorating; abatement; remission; diminishing of symptoms or delaying the onset of symptoms; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; and / or improving a subject's physical or mental well-being or reversing a sign or symptom of a health condition, disease or disorder to any extent, and includes, but does not require, a complete cure of the condition, disease, or disorder. Treating can be curing, improving, or partially ameliorating a disorder. “Treatment” can also include improving or enhancing a condition or characteristic, for example, bringing the function of a particular system in the body to a heightened state of health or homeostasis.

[0039] As used herein, “preventing” a health condition, disease, or disorder refers to avoiding, delaying, forestalling, or minimizing the onset of a particular sign or symptom of the condition, disease, or disorder. Prevention can, but is not required, to be absolute or complete; meaning, the sign or symptom may still develop at a later time. Prevention can include reducing the severity of the onset of such a condition, disease, or disorder, and / or inhibiting the progression of the condition, disease, or disorder to a more severe condition, disease, or disorder.

[0040] In some embodiments of the invention, the method comprises administration of multiple doses of the compounds of the subject invention. The method may comprise administration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 or more therapeutically effective doses of a composition comprising the compounds of the subject invention as described herein. In some embodiments, doses are administered over the course of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days, 21 days, 30 days, 2 months, 3 months, 6 months, 9 months, 1 year, 1.5 years, 2 years, 2.5 years, 5 years or more than 10 years. The frequency and duration of administration of multiple doses of the compositions is such as prevent or treat anemia. Moreover, treatment of a subject with a therapeutically effective amount of the compounds of the invention can include a single treatment or can include a series of treatments. It will also be appreciated that the effective dosage of a compound used for treatment may increase or decrease over the course of a particular treatment. Changes in dosage may result and become apparent from the results of testing for a neurodegenerative disorder. In some embodiments of the invention, the method comprises administration of the compounds at several time per day, including but not limiting to 2 times per day, 3 times per day, and 4 times per day.

[0041] As used herein, an “isolated” or “purified” compound is substantially free of other compounds. In certain embodiments, purified compounds are at least 60% by weight (dry weight) of the compound of interest. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 99%, by weight of the compound of interest. For example, a purified compound is one that is at least 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99%, or 100% (w / w) of the desired compound by weight. Purity is measured by any appropriate standard method, for example, by column chromatography, thin layer chromatography, or high-performance liquid chromatography (HPLC) analysis.

[0042] By “reduces” is meant a negative alteration of at least 1%, 5%, 10%, 25%, 50%, 75%, or 100%.

[0043] By “increases” is meant as a positive alteration of at least 1%, 5%, 10%, 25%, 50%, 75%, or 100%.

[0044] As used herein, a “pharmaceutical” refers to a compound manufactured for use as a medicinal and / or therapeutic drug.

[0045] As used herein, “subject”, “host” or “organism” refers to any member of the phylum Chordata, more preferably any member of the subphylum vertebrata, or most preferably, any member of the class Mammalia, including, without limitation, humans and other primates, including non-human primates such as rhesus macaques, chimpanzees and other monkey and ape species; livestock, such as cattle, sheep, pigs, goats and horses; domestic mammals, such as dogs and cats; laboratory animals, including rabbits, mice, rats and guinea pigs; birds, including domestic, wild, and game birds, such as chickens, turkeys, ducks, and geese. The term does not denote a particular age or gender. Thus, adult, young, and new-born individuals are intended to be covered as well as male and female subjects. In some embodiments, a host cell is derived from a subject (e.g., tissue specific cells or stem cells). In some embodiments, the subject is a non-human subject. A “patient” or “subject” receiving the composition or treatment method of this invention is a human, typically adult human, of any age, gender, and ethnic background, who has been diagnosed to suffer from a neurodegenerative disorder, exhibiting one or more pertinent symptoms, or who has been identified as having increased risk of developing a neurodegenerative disorder due to family history and / or known genetic mutations even though the subject may not yet develop any symptoms of the disorder. Typically, a patient diagnosed with or at risk of developing a neurodegenerative disorder is at least about 60, about 70 or about 80 years old. In some early onset cases, however, such a subject is at least about 35, about 40, about 50 or about 60 years old.

[0046] The term "inhibiting" or "inhibition," as used herein, refers to any detectable negative effect on a target biological process, such as, for example, any neurodegenerative symptoms exhibited by a patient suffering from a neurodegenerative disorder, such as, for example, Alzheimer's disease (AD). Typically, an inhibition of a neurodegenerative symptom is reflected in a decrease of at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90% or higher, including 100% or complete elimination, of one or more hallmarks of a neurodegenerative disorder (e.g., AD), as described herein, when compared to a control not given the “inhibition” treatment, such as treatment by administration of small molecule therapeutics described herein, on the other hand, inhibition of a neurodegenerative symptom may also be manifested as increased cell survival (e.g., neuronal cell survival), demonstrated in an increase of at least about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 200%, about 300%, about 500% or more in the number or length of time of cell survival in the pertinent tissues, including, for example, in the central nervous system (CNS) within the subject after the administration of the PTEN:PDZ interaction inhibitors in comparison to a control that has not received the PTEN:PDZ interaction inhibitors.

[0047] As used herein, the phrase “isochlorogenic acids” is a generic term, which also refers to dicaffeoylquinic acids, including, for example, 3,5-dicaffeoylquinic acid, 3,4- dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid, wherein the structure of described isochlorogenic acid B (Formula (I)) and isochlorogenic C (Formula (II)) is as follows: Formula (I)

[0048] As used herein, the term “amaranth” refers to small molecule compound 3 -Hydroxy -4- [(4-sulfonatonaphthalen-l-yl)diazenyl]naphthalene-2,7-disulfonate trisodium salt (Formula (III)) It has the following chemical structure:

[0049] Formula (III)

[0050] As used herein, the term “pentagalloylglucose” refers to small molecule compound also known as [(2R,3R,4S,5R,6S)-3,4,5,6-tetrakis[(3,4,5-trihydroxybenzoyl)oxy] oxan-2- yl]methyl 3,4,5-trihydroxybenzoate (Formula (IV)). It has the following chemical structure: Formula (IV)

[0051] As used herein, the term “hematoporphyrin dihydrochloride” refers to small molecule compound also known as 3-[18-(2-carboxyethyl)-8,13-bis(l-hydroxyethyl)-3,7,12,17- tetramethyl-22,23-dihydroporphyrin-2-yl]propanoic acid;dihydrochloride ((Formula (V)). It has the following chemical structure:

[0052] Formula (V)

[0053] As used herein, the term “neurodegenerative disorder” includes, but is not limited to, the following conditions: (1) Diseases of central motor systems including degenerative conditions affecting the basal ganglia (Huntington's disease, Wilson's disease, striatonigral degeneration, corticobasal ganglionic degeneration), Tourette’s syndrome, Parkinson’s disease, progressive supranuclear palsy, progressive bulbar palsy, familial spastic paraplegia, spinomuscular atrophy, amyotrophic lateral sclerosis (ALS) and variants thereof, dentatorubral atrophy, olivo-pontocerebellar atrophy, paraneoplastic cerebellar degeneration, and dopamine toxicity; (2) Diseases affecting sensory neurons such as Friedreich's ataxia, diabetes, peripheral neuropathy, retinal neuronal degeneration; (3) Diseases of limbic and cortical systems such as cerebral amyloidosis, Pick's atrophy, Retts syndrome; (4) Neurodegenerative pathologies involving multiple neuronal systems and / or brainstem including Alzheimer's disease, AIDS- related dementia, Leigh's disease, diffuse Lewy body disease, epilepsy, multiple system atrophy, Guillain-Barre syndrome, lysosomal storage disorders such as lipofuscinosis, late- degenerative stages of Down's syndrome, Alper's disease, vertigo as result of central nervous system (CNS) degeneration, corticobasal degeneration and progressive supranuclear palsy; (5) Pathologies associated with developmental retardation and learning impairments, Down's syndrome, and oxidative stress induced neuronal death; (6) Pathologies arising with aging and chronic alcohol or drug abuse including, for example, with alcoholism the degeneration of neurons in locus coeruleus, cerebellum, cholinergic basal forebrain; with aging degeneration of cerebellar neurons and cortical neurons leading to cognitive and motor impairments; and with chronic amphetamine abuse degeneration of basal ganglia neurons leading to motor impairments; (7) Pathological changes resulting from focal trauma such as stroke, focal ischemia, vascular insufficiency, hypoxic-ischemic encephalopathy, hyperglycemia, hypoglycemia, closed head trauma, or direct trauma; (8) Pathologies arising as a negative sideeffect of therapeutic drugs and treatments (e.g., degeneration of cingulate and entorhinal cortex neurons in response to anticonvulsant doses of antagonists of the N-methyl-D-aspartate (NMDA) class of glutamate receptor); and (9) Clinical diagnosis associated with depressive disorder covering, for example, major depression, persistent depressive disorder, bipolar disorder, seasonal affective disorder, psychotic depression, postpartum depression, premenstrual dysphoric disorder, situational depression, atypical depression, and treatment resistant depression.

[0054] As used herein, the term “Alzheimer's Disease” or AD refers to a chronic neurodegenerative disease with a progressive pattern of cognitive and functional impairment. Although certain genetic mutations have been identified as contributing to the risk of developing AD, the exact cause of the disease remains largely undetermined. On the other hand, the symptoms of AD are well-recognized: initially a patient experiences difficulty in remembering recent events, i.e., short-term memory loss. As the disease develops, the patient further exhibits symptoms such as difficulties with language, disorientation, mood swings, loss of motivation, inability of self-care, and behavioral problems. As the patient's condition declines, he typically begins to withdraw from family and society. Gradually, the patient's bodily functions are lost, which ultimately leads to death. Although the pace of disease progression can vary, the typical life 30 expectancy of an AD patient is around three to nine years following diagnosis. AD usually affects patients aged 65 years or older, with about 5% of AD diagnosis made in patients younger than 65.

[0055] The terms “therapeutically effective amount” or “effective amount,” as used herein, refers to an amount that produces therapeutic effects for which a substance is administered. The effects include the prevention, correction, or inhibition of progression of the symptoms of a disease / condition and related complications to any detectable extent, e.g., one or more of the hallmarks of a neurodegenerative disorder (e.g., AD). The exact amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); and Pickar, Dosage Calculations (1999)).

[0056] A "pharmaceutically acceptable" or "pharmacologically acceptable" excipient is a substance that is not biologically harmful or otherwise undesirable, i.e., the excipient may be administered to an individual along with a bioactive agent without causing any undesirable biological effects. Neither would the excipient interact in a deleterious manner with any of the components of the composition in which it is contained.

[0057] The term "excipient" refers to any essentially accessory substance that may be present in the finished dosage form of the composition of this invention. For example, the term "excipient" includes vehicles, binders, disintegrants, fillers (diluents), lubricants, glidants (flow enhancers), compression aids, colors, sweeteners, preservatives, suspending / dispersing agents, film formers / coatings, flavors and printing inks.

[0058] An "antibody" refers to a polypeptide substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, which specifically bind and recognize an analyte (antigen). The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD, and IgE, respectively.

[0059] An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chains respectively.

[0060] Antibodies exist, e.g., as intact immunoglobulins or as a number of well characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)'2, a dimer of Fab which itself is a light chain joined to VH-CH1 by a disulfide bond. The F(ab)'2 may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)'2 dimer into an Fab' monomer. The Fab' monomer is essentially an Fab with part of the hinge region (see, Paul (Ed.) Fundamental Immunology, Third Edition, Raven Press, NY (1993)). While various antibody fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that such fragments may be synthesized de novo either chemically or by utilizing recombinant DNA methodology.

[0061] Further modification of antibodies by recombinant technologies is also well known in the art. For instance, chimeric antibodies combine the antigen binding regions (variable regions) of an antibody from one animal with the constant regions of an antibody from another animal. Generally, the antigen binding regions are derived from a non-human animal, while the constant regions are drawn from human antibodies. The presence of the human constant regions reduces the likelihood that the antibody will be rej ected as foreign by a human recipient. On the other hand, "humanized" antibodies combine an even smaller portion of the non-human antibody with human components. Generally, a humanized antibody comprises the hypervariable regions, or complementarity determining regions (CDR), of a non-human antibody grafted onto the appropriate framework regions of a human antibody. Antigen binding sites may be wild type or modified by one or more amino acid substitutions, e.g., modified to resemble human immunoglobulin more closely. Both chimeric and humanized antibodies are made using recombinant techniques, which are well-known in the art (see, e.g., Jones et al. (1986) Nature 321 :522-525).

[0062] Thus, the term "antibody," as used herein, also includes antibody fragments either produced by the modification of whole antibodies or antibodies synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv, a chimeric or humanized antibody).

[0063] Compounds and Methods for Inhibiting the PTEN:PDZ interaction The subject invention provides a novel and effective method for treating neurodegenerative disorders, in which one or more neurodegenerative symptoms result from or are exacerbated by Ap-mediated interaction between PTEN and PSD-95.

[0064] Provided are compounds and compositions for inhibiting the PTEN:PDZ interaction in a subject. Advantageously, the compositions of the invention comprise isochl orogenic acid B

[0065] (Formula (I)), isochlorogenic C (Formula (II)), amaranth (Formula (HI)), pentagalloylglucose (Formula (IV)), hematoporphyrin dihydrochloride ((Formula (V)), or any combination thereof:

[0066] Formula (I)

[0067] Formula (III)

[0068] Formula (IV)

[0069] Formula (V) Also provided are methods of using the compounds and compositions of the invention to treat neurodegenerative disorders, including, but not limited to, AD.

[0070] The skilled artisan will understand that the dosage of the compositions of the instant invention varies, depending upon, for example, the route of administration, other drugs being administered, and the age, condition, gender and seriousness of the disease in the subject as described above. An effective dose of a isochlorogenic acid B, isochl orogenic C, amaranth, pentagalloylglucose, hematoporphyrin dihydrochloride, or any combination thereof generally ranges between about 0.001 pg / kg of body weight and 100 mg / kg of body weight. Examples of such dosage ranges include, but are not limited to, about 0.01 pg / kg to about 100 mg / kg; about 0.1 mg / kg to about 75 mg / kg; about 1 mg / kg to about 50 mg / kg; about 5 mg / kg to about 25 mg / kg; or about 10 mg / kg to 20 mg / kg. In certain embodiments, the subject compounds can be administered at least about one, two, three or more times per day. In certain embodiments, the subject compounds can be administered daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, weekly, every 8 days, every 9 days, every 10 days, every 11 days, every 12 days, every 13 days, biweekly (z.e., every other week), monthly, bimonthly (z.e., every other month), every 3 months, every 6 months, or every year. In certain embodiments, the treatment can be administered over a period of at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, about 13 days, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 3 months, about 6 months, about 9 months, about 1 year, about 18 months, about 2 years, about 3 years, about 4 years, about 5 years, or about 10 years. In preferred embodiments, the composition of the subject invention can be administered at 10 mg / kg per day for about 13 days. In preferred embodiments, the composition can also be administered at 20 mg / kg per day for 13 days.

[0071] The method of the invention can be practiced by administering the composition according to any administration route capable of providing a therapeutic effect in the subject. Suitable administration routes include, but are not limited to orally, topically, buccally, sublingually, parenterally, intravenously, intraarterially, subdermally, subcutaneously, ocularly, intravaginally, rectally, by inhalation, or a combination thereof. The composition can be administered as a powder, liquid, pill, tablet, pellet, capsule, thin film, solution, spray, syrup, linctus, lozenge, pastille, chewing gum, paste, vapor, suspension, emulsion, ointment, cream, lotion, liniment, gel, drop, topical patch, buccal patch, bead, gummy, gel, sol, or injection.

[0072] In some embodiments, the therapeutically effective amount of isochlorogenic acid B, isochlorogenic C, amaranth, pentagalloylglucose, hematoporphyrin dihydrochloride, or any combination thereof can be administered through intravenous or subcutaneous administration or in a form suitable for administration by inhalation or insufflation, including powders and liquid aerosol administration, or by sustained release systems such as semipermeable matrices of solid hydrophobic polymers containing the compounds of the invention. Administration may be also by way of other carriers or vehicles such as patches, micelles, liposomes, vesicles, implants (e.g., microimplants), synthetic polymers, microspheres, nanoparticles, and the like. In certain embodiments, the coordination compound compositions may be administered using a nanoparticle to passage the composition through skin for treating peripheral (close to skin) conditions.

[0073] In some embodiments, the at least one compositions of the instant invention may be formulated for parenteral administration e.g., by injection, for example, bolus injection, intravenous administration, or continuous infusion. In addition, the compositions may be presented in unit dose form in ampoules, pre-filled syringes, and small volume infusion or in multi-dose containers with or without an added preservative. The compositions may be in forms of suspensions, solutions, or emulsions in oily or aqueous vehicles. The composition may further contain formulation agents such as suspending, stabilizing and / or dispersing agents. In further embodiments, the active ingredients of the compositions according to the instant invention may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

[0074] The subject compositions can further comprise one or more pharmaceutically acceptable carriers, and / or excipients, and can be formulated into preparations, for example, semi-solid or liquid forms, such as solutions or injections.

[0075] The term “pharmaceutically acceptable” as used herein means compatible with the other ingredients of a pharmaceutical composition and not deleterious to the recipient thereof.

[0076] Carriers and / or excipients according to the subject invention can include any and all solvents, diluents, buffers (such as, e.g. , neutral buffered saline, phosphate buffered saline, or optionally Tris-HCl, acetate or phosphate buffers), oil-in-water or water-in-oil emulsions, aqueous compositions with or without inclusion of organic co-solvents suitable for, e.g, IV use, solubilizers (e.g., Polysorbate 65, Polysorbate 80), colloids, dispersion media, vehicles, fillers, chelating agents (e.g, EDTA or glutathione), amino acids (e.g., glycine), proteins, disintegrants, binders, lubricants, wetting agents, emulsifiers, sweeteners, colorants, flavorings, aromatizers, thickeners (e.g., carbomer, gelatin, or sodium alginate), coatings, preservatives e.g., Thimerosal, benzyl alcohol, polyquaterium), antioxidants (e.g., ascorbic acid, sodium metabisulfite), tonicity controlling agents, absorption delaying agents, adjuvants, bulking agents (e.g., lactose, mannitol) and the like. The use of carriers and / or excipients in the field of drugs and supplements is well known. Except for any conventional media or agent that is incompatible with the target health-promoting substance or with the composition, carrier or excipient use in the subject compositions may be contemplated.

[0077] In certain embodiments, a patient is suitable for receiving the treatment method of this invention is a person who has been diagnosed with a neurodegenerative disorder such as AD and exhibits some pertinent symptoms, or who has not yet been given the diagnosis of a neurodegenerative disorder and does not exhibit any symptoms but has been deemed to be at increased risk of developing a neurodegenerative disorder such as AD at a later time due to family history and / or known genetic risks. In some cases, the suitable subject may be younger than 60 or 65, e.g., in the age range of about 30-40,40-50,50-60, or 60-64. Among the known genetic risk factors, genes encoding the amyloid precursor protein (APP) and presenilins 1 and 2 have been found to exhibit a high rate of mutation in familial AD. Another genetic risk for AD is mutations in the TREM2 (triggering receptor expressed on myeloid cells 2) gene, which are reported as increasing the likelihood of one developing AD by 3-5 fold. One further known genetic risk factor is the inheritance of the 84 allele of the apolipoprotein E (APOE) gene, with about 40-80% AD patients have at least one copy of this allele. More recent genome-wide association studies (GWAS) have found 19 areas in genes that appear to affect the risk of developing AD. These genes include: CASS4, CELF1, FERMT2, HLA-DRB5, INPP5D, MEF2C, NME8, PTK2B, SORL1, ZCWPW1, SIC24A4, CPU, PICALM, CR1, BINI, MS4A, ABCA7, EPHA1, and CD2AP. Thus, genetic analysis of these relevant genomic loci can aid identification of subjects at a heightened risk for AD that may be suitable recipients for the treatment method of the present invention.

[0078] In certain embodiments, the composition of the subject invention can inhibit the interactions between PTEN and PSD-95 and exhibit low cytotoxicity.

[0079] In certain embodiments, the composition of the subject invention can protect cells against amyloid beta-induced cell death.

[0080] In certain embodiments, the invention provides a method for treating neurodegenerative disorders in a subject in need thereof comprising administering to the subject a composition comprising amaranth, isochlorogenic acid B, isochl orogenic C, pentagalloylglucose, or hematoporphyrin dihydrochloride. The composition can comprise one of more of the compounds described herein. The method can prevent AD in a subject at risk of developing Alzheimer’s disease and / or alleviate ongoing AD symptoms.

[0081] In certain embodiments, the subject invention provides kits for treating a neurodegenerative disorder, such as, for example, AD, according to the method of the present invention. In certain embodiments, the kit can include a first container that contains a pharmaceutical composition comprising isochl orogenic acids, such as, for example, isochlorogenic acid C and / or isochlorogenic acid B, and / or amaranth and at least one pharmaceutically acceptable carrier and / or excipient, optionally with a second container that contains a pharmaceutical composition comprising another therapeutically effective compound, such as, for example, donepezil, or ameliorating the symptoms of the neurodegenerative disorder. In some embodiments of the kits, a single container may contain a pharmaceutical composition comprising two or more of compounds effective for treating a neurodegenerative disorder (e.g., AD or depression). In certain embodiments, compounds and compositions that directly modulate the interactions of PTEN with PDZ proteins can be used to treat depression. The kits may further include informational material providing instructions on how to dispense the pharmaceutical composition(s), including description of the type of subject who may be treated (e.g., human patients who have received a diagnosis of a neurodegenerative disorder (e.g., AD or depression) or have been deemed as with high risk of developing the neurodegenerative disorder including AD, for example, due to a strong propensity indicated by a family history or genetic risk factor), the schedule (e.g., dose and frequency) and route of administration, and the like.

[0082] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

[0083] Following are examples that illustrate procedures for practicing the invention. These examples should not be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise noted.

[0084] EXAMPLE 1— FLUORESCENT-BASED BINDING ASSAY THAT MONITOR THE INTERACTION BETWEEN PTEN PDZ-BM AND PSD-95 / MAGI2 UPON TREATING OF 206 CANDIDATE COMPOUNDS. A vast library of 100,000 compounds was screened through advanced computational techniques, such as Quantitative Structure- Activity Relationship (QSAR) modeling and molecular docking studies that systematically reduced to 1205 compounds. Further computational filtering using PT-QSAR model allowed for the prediction of drug-target interactions by analyzing molecular descriptors like Polar Surface Area (PSA) and Log P. This approach resulted in 206 compounds that were most likely to mimic the PTEN-PDZ peptide interaction with PSD95, a key focus for Alzheimer’s treatment.

[0085] The compounds were prepared in water, ethanol, pyridine or dimethyl sulfoxide (DMSO) and further diluted for single dose screen and IC50 values estimation accordingly.

[0086] Screening for small molecules disrupting PTEN:PSD-95 / MAGI2 interaction

[0087] The binding affinity between fluorescein-labeled PTEN PDZ-BM peptide and PSD- 95 / MAGI2 was measured by the fluorescence polarization (FP) assay. It is a biochemical assay for measuring the affinity of the interaction between a small ligand and its interacting protein of a greater mass. Fluorescent-labeled ligands in solution have a scattered emission profile when excited. When the ligand binds to a bulky protein partner, it will cause “polarization” of the fluorescent emission, which can be detected by a spectrophotometer (Moerke 2009).

[0088] GST-PSD-95 PDZ2 and GST-MAGI2 PDZ2 were expressed by bacteria and purified. The fluorescent-labeled peptide FITC-PFDEDQHTQITKV-COOH (SEQ ID NO: 1) was synthesized and used as a fluorescent probe. FP assay was performed by mixing increasing concentration of GST-PSD-95 PDZ2 and GST-MAGI2-PDZ2 fusion proteins with fixed 5 nM fluorescent probe for 30 min in 25 pL of TBS buffer per well of a 384-well plate in triplicate. The polarization value was measured by SpectraMax i3® spectrophotometer. The binding curves were shown by plotting concentration against the Polarization values (mP).

[0089] The results set out in FIGs. 1A-1B showed that 20 pM and 40 pM of MAGI2-PDZ2 and PSD95-PDZ2 were able to produce -80% of the increase in polarization value, respectively, and therefore were selected to validate the inhibitory effects of the 206 candidate compounds using a single drug dose of 200 pM. 23 compounds displayed inhibitory effects (FIGs. 2A-2B)

[0090] Estimation of IC50 values

[0091] The IC50 values (concentration of test compounds that inhibits the PTEN and PSD-95 binding by 50%) were determined by FP assay. 40 pM GST-PSD95-PDZ2 fusion protein was incubated with 5 nM fluorescent probe in the absence or presence of increasing concentrations of 23 candidate compounds. Competitive binding curves of 11 candidate compounds with the lowest IC50 values were shown in FIGs. 3A-3K. IC50 values were obtained from dose-effect curves by linear regression.

[0092] EXAMPLE 2— INHIBITORY EFFECTS OF PROMISING COMPOUNDS IN BLOCKING THE PHYSICAL BINDING BETWEEN PTEN AND PSD-95 IN GST PULL DOWN ASSAY AND CO-IMMUNOPRECIPITATION ASSAY

[0093] Determine the efficacy of candidate compounds in blocking PTEN binding to PSD-95

[0094] The ability of candidate compounds (IC5CX60 pM) to block the physical interaction between PTEN and PSD-95 are validated by GST pull down assay and co-immunoprecipitation assay. For GST pull-down assay, 293T cells were transfected with 8 pg PTEN WT full length vector. Two days later the cells were lysed with ice-cold lysis buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% NP-40, 1 mM EDTA and protease inhibitors). Supernatants were stored in -80°C. until further use. Purified GST-PSD- 95-PDZ2 protein were re-incubated with glutathione-sepharose beads at 4°C for at least 4 hr and washed twice with GST wash buffer (10 mM Tris-HCl pH 7.5, 0.2% Triton X-100, 150 mM NaCl) and twice with PBS. After the washes, the beads were incubated with 200 pM compound for 30 min then incubated with collected supernatants (-500 pg) for another 1 hr. Samples were washed twice with PBS, twice with GST wash buffer and once with lysis buffer and boiled with Laemmli sample buffer. The binding complexes was resolved by SDS-PAGE and analyzed by western blot and Coomassie blue staining.

[0095] For co-immunoprecipitation assay, 293T cells were transfected with 3 pg full length AU5-PTEN 3 A vector (with three phosphorylation sites mutated to As and thereby open up the PTEN structure for better PDZ binding) and full length PSD95 vector. 48 hours after the transfection, cells were treated with candidate compounds for 2 hours and then lysed with ice- cold lysis buffer. Co-immunoprecipitation were performed by incubating 0.5-1 mg protein extracts with AU5 antibody or IgG (control) at 4°C overnight. The GammaBind G Sepharose beads were added to incubate for another 4-6 hours at 4°C. The samples were then washed and boiled with Laemmli sample buffer. Immunoprecipitated proteins were separated by 10% SDS- PAGE.

[0096] The remarkable inhibition effects of #58 amaranth, #82 isochlorogenic acid C and #84 isochl orogenic B were observed in the western blot analysis results in FIGs. 4A-4B. EXAMPLE 3— EVALUATING NEUROTOXICITY AND NEUROPROTECTION BY MTS ASSAY AND IPSC-NEURON CULTURE ASSAY.

[0097] The sample was prepared in dimethyl sulfoxide (#82 isochl orogenic acid C, #84 isochlorogenic acid B, #80 pentagalloylglucose and #148 hematoporphyrin dihydrochloride) and in PBS (#58 amaranth) and further diluted to 0, 12.5, 25, 50, 100, 200, 400 pM for in vitro studies.

[0098] In vitro cell viability assays

[0099] The toxicity of #82 isochlorogenic C, #84 isochlorogenic B, #58 amaranth and #80 pentagalloylglucose and #148 hematoporphyrin dihydrochloride was tested in iPSC-derived NPCs (FIG. 5A) and iPSC-derived neurons (FIG. 5B). Cells were seeded in 96-well plate and treated with serial concentrations of #82 isochlorogenic acid C, #84 isochlorogenic acid B, #58 amaranth and #80 pentagalloylglucose, #148 hematoporphyrin dihydrochloride and vehicle (control) for 24 hours (iPSC-NPC) or 48 hours (iPSC-neuron). Cell viability was measured using a [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium] (MTS) reductio assay. MTS / PMS (20: 1) solution was mixed with phenol red free culture medium immediately prior to the addition to the cell cultures at a ratio of 1 :5 (MTS / PMS: culture medium). The plates were incubated at 37°C, and the absorbance was detected at 490 nm wavelength by plate reader. The data are reported as Mean ± SD.

[0100] In vitro binding assays

[0101] Effect of #82 isochlorogenic C, and #84 isochlorogenic B, on the interaction between PTEN and PSD-95 in iPSCs. The effect of AP1-42 was determined by incubating 10 pM Ap42 with iPSC-NPCs for 24 hours and co-immunoprecipitation was performed to confirm the increased association between PSD95 and PTEN (FIG. 6A). PSD95:PTEN binding induced by AP42 in iPSC-neuron was partially abolished by #82 isochlorogenic C, and #84 isochlorogenic B treatment (FIG. 6B).

[0102] Evaluation of neuroprotection iPSC-derived neurons were treated with 100 pM #82 + 100 pM #84, 200 pM #148 or vehicle (2% DMSO) for 2 hours and then treated with 10 pM AP1-42 or NEk’ELO vehicle (0.005%) for 48 hours. Bright-field images were taken at 24 hours and 48 hours. Representative images of iPSC-neurons were shown in FIG. 7. The results showed that #82 isochlorogenic C and #84 isochl orogenic B protected against APi-42-induced neuronal cell death.

[0103] In vivo interaction between PTEN and PSD-95

[0104] The endogenous protein expressions of PTEN, PSD95, pERK and ERK were analyzed by Western blot (FIG. 8A) and the association between PTEN and PSD95 was assessed by coimmunoprecipitation in synaptosomes collected after behavioral tests in WT and 5XFAD mice treated with #82 isochlorogenic C, and #84 isochlorogenic B (FIG. 8B).

[0105] EXAMPLE 4— RESCUING THE COGNITIVE SYMPTOMS OF ALZHEIMER’S DISEASE. THE EFFECT OF COMPOSITION WAS EVALUATED BY MORRIS WATER MAZE TASK.

[0106] Mouse Strains: (A)Wild-type (WT): C57BL / 6J (Stock Number 000664, The Jackson Laboratory). (B)5XFAD: B6.Cg-Tg(APPSwFlLon, PSEN1*M146L*L286V) 6799Vas / J (Stock Number 5008730, The Jackson Laboratory).

[0107] Gender: Male

[0108] Number of animals: 99

[0109] Housing: ^5 / cage, ~18-23°C with 40-60% humidity on a 12-hours light / dark cycle.

[0110] 5xFAD mice

[0111] 5XFAD transgenic mice overexpress mutant human App (695) with the Swedish (K670N, M671L), Florida (1716V), and London (V717I) Familial Alzheimer's Disease (FAD) mutations along with human Psenl harboring two FAD mutations, M146L and L286V. Both transgenes are regulated by the mouse Thyl promoter to drive overexpression in the brain (Oakley et al., 2006). This mouse line is propagated by crossing hemizygous mice with C57BL / 6J. Only male mice approximately 7-8 months of age are used for experimentation.

[0112] Experimental design

[0113] The small molecule compound, #58 amaranth, #82 isochlorogenic acid C, #84 isochlorogenic acid B, #80 pentagalloylglucose, and #148 hematoporphyrin dihydrochloride were tested for therapeutic efficacy to alleviate memory deficits in 5xFAD transgenic mouse strain, respectively. The groups and treatment of the composition is detailed in Table 1.

[0114] TABLE 1

[0115] Experimental design for the evaluation of cognitive function in mice The mice were given the inventive composition every morning as Table 1 instructed.

[0116] The Morris water maze was performed on the 8th day and lasted for 6 days. The study parameters included escape latency, percentage of time spent in the target quadrant, and number of entries to the target zone. Morri s W ater Maze Te st

[0117] The Morris water maze (MWM) task is a well-established cognitive test aiming to assess the hippocampal-dependent learning and spatial memory (Morris 1981). The detail technical procedures are as follows:

[0118] I. Mice were pre-handled daily for one week before tests and transferred from the holding room to the behavior room 30 min prior to the beginning of the test. Mice were placed in a place where they cannot see the experimental equipment. II. Equipment preparation: A round and white tank with a diameter of 150 cm and a depth of 76 cm. The tank was surrounded by black curtains. Four spatial cues were placed above the water surface at equal distance from each other. A 10-cm diameter circular platform in clear plexiglass was placed in the tank.

[0119] III. The pool was filled with water until the water surface was slightly above the platform. The water was equilibrated to room temperature (22°C) and colored by white and nontoxic paint (Washable paint, Crayola) to hide the platform from day 2.

[0120] IV. Pre-training (Day 0, 8th injection day): A red flag was placed on the platform to increase its visibility. Starting directions for each trial were preset. Mice were gently put into the tank with the face to the wall, make sure they could not see the spatial cues. If the mice find the platform within 60 seconds, they were allowed to stay on the platform for 5 seconds and then returned to the cage. If the mice failed to find the platform within 60 seconds, they were placed on the platform for 20 seconds before returning to its cage. Each mouse has five trails per day with different starting direction. Mice were dried off before returning to animal facility.

[0121] V. Training session (Days 1-4, 9th- 12th injection day): The flag was removed and nontoxic, white paint was added to hide the platform. Handling of the mice were same as Day 0.

[0122] VI. Probe Trial (Day 5, 13th injection day): The platform was removed from the tank. Mice were only examined once and departed from the furthest starting point to the platform for a duration of one minute.

[0123] Image Capture and Processing

[0124] Video images were captured with a Logitech C31 video camera system using the QuickTime Player Applications. Video files were imported and analyzed by the Etho Vision® XT software (Noldus). Software settings: The tank was calibrated so the software can calculate actual distance information from pixel information. The 4 quadrants and 5 platform subzonesone in each quadrant, and one in the center of the pool were defined and entitled in the software. Tracking: Dynamic differencing mode was used to track the swimming path.

[0125] Data Analysis

[0126] For pre-training and training sessions, the averages of the parameters from 5 trials were calculated to give a single escape latency on each day for each mouse. For day 6, single value of escape latency and number of entries to the platform were collected. The data were analyzed using one-way and two-way ANOVA.

[0127] RESULTS

[0128] Statistical analysis of results derived from pre-training day revealed that there was no difference in visual and motor ability among the five experimental groups (FIG. 9A). Importantly, during 4 days of training (Days 1-4), #58 amaranth, #84 isochl orogenic acid B, #82 isochl orogenic C, #80 pentagalloylglucose, or #148 hematoporphyrin dihydrochloride treated 5xFAD showed a reduced latency time compared with 5xFAD control at Day4 (Two- way ANOVA, @ / ?<0.05, ### / ?<0.001 and#### / ?<0.0001 compared with 5xFAD control group,+p<0.01 and ° / op<0.01 compared with WT control group) (FIG. 9B) Moreover, more entries into the platform area and shorter latency to their first swim across the platform area were observed in treated 5xFAD during the probe trial (FIGs. 9C-9D) (One-way ANOVA: These findings suggest that the present inventive composition has therapeutic effects on memory deficits in 5XFAD mice. These results provide a new therapeutic target for P-amyloid associated memory deficits and the inventive composition can serve as a treatment for Alzheimer's disease.

[0129] All patents, patent applications, and other publications, including GenBank Accession Numbers, cited in this application are incorporated by reference in the entirety for all purposes.

[0130] It should be understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims. In addition, any elements or limitations of any invention or embodiment thereof disclosed herein can be combined with any and / or all other elements or limitations (individually or in any combination) or any other invention or embodiment thereof disclosed herein, and all such combinations are contemplated with the scope of the invention without limitation thereto. EXEMPLARY EMBODIMENTS

[0131] Embodiment 1. A method for inhibiting an interaction of phosphatase and tensin homologue deleted from chromosome 10 (PTEN) and PSD95 / Disc large / Zonula occludens-1 (PDZ), comprising administering to a subject in need thereof a composition comprising an effective amount of at least two compounds selected from amaranth, pentagalloylglucose, isochlorogenic acid C, isochl orogenic acid B, and hematoporphyrin dihydrochloride.

[0132] Embodiment 2. The method of embodiment 1, wherein the composition comprises isochlorogenic acid B and isochlorogenic acid C; amaranth and isochlorogenic acid C; isochlorogenic acid B and amaranth; or isochlorogenic acid B, isochlorogenic acid C, and amaranth.

[0133] Embodiment 3. The method of any preceding embodiments, wherein the subject has a neurodegenerative disorder.

[0134] Embodiment 4. The method of embodiment 3, wherein the neurodegenerative disorder is Alzheimer’s Disease.

[0135] Embodiment 5. The method of any preceding embodiments, wherein the administering comprises injecting the composition into the subject.

[0136] Embodiment 6. The method of embodiment 5, wherein the injecting is by intravenous, intramuscular, intraperitoneal, or subcutaneous injection.

[0137] Embodiment 7. The method of any preceding embodiments, wherein the administering comprises oral ingestion.

[0138] Embodiment 8. The method of any preceding embodiments, wherein the administering comprises nasal inhalation. Embodiment 9. The method of any preceding embodiments, wherein the isochlorogenic acid B, isochl orogenic acid C, and / or amaranth are administered in an amount of about 5 mg / kg to about 100 mg / kg subject body weight.

[0139] Embodiment 10. The method of embodiment 9, wherein the isochlorogenic acid B, isochlorogenic acid C, and / or amaranth are administered in an amount of about 10 mg / kg to about 20 mg / kg subject body weight.

[0140] Embodiment 11. The method of embodiment 10, wherein the isochlorogenic acid B and / or isochlorogenic acid C are administered in an amount of about 20 mg / kg subject body weight.

[0141] Embodiment 12. The method of embodiment 10, wherein the amaranth is administered in an amount of about 10 mg / kg subject body weight.

[0142] Embodiment 13. The method of any preceding embodiments, wherein the pentagalloylglucose and / or hematoporphyrin dihydrochloride are administered in an amount of about 10 mg / kg subject body weight.

[0143] Embodiment 14. The method of embodiment 11, wherein the isochlorogenic acid B and / or isochlorogenic acid C are administered in an amount of about 20 mg / kg subject body weight and the amaranth is administered in an amount of about 10 mg / kg.

[0144] Embodiment 15. A composition comprising at least two compounds selected from amaranth, pentagalloylglucose, isochlorogenic acid C, isochlorogenic acid B, and hematoporphyrin dihydrochloride; and a pharmaceutically effective carrier and / or excipient.

[0145] Embodiment 16. The composition of embodiment 15, further comprising a therapeutic agent effective for treating a neurodegenerative disorder.

[0146] Embodiment 17. A kit comprising a first container containing a pharmaceutical composition comprising at least two compounds selected from amaranth, pentagalloylglucose, isochl orogenic acid C, isochl orogenic acid B, and hematoporphyrin dihydrochloride, and a second container containing a therapeutic agent effective for treating a neurodegenerative disorder. Embodiment 18. The kit of embodiment 17, wherein the neurodegenerative disorder is Alzheimer's Disease.

[0147] Embodiment 19. The kit of embodiment 17, further comprising instructions for administration of the contents of the first container and the contents of the second container to a subject.

[0148] REFERENCES

[0149] E Alizadeh, M., Demir, E., Aydogdu, N., Zare, N., Karimi, F., Kandomal, S. M., Rokni, H., and Ghasemi, Y. (2022). Recent advantages in electrochemical monitoring for the analysis of amaranth and carminic acid food colors. Food and Chemical Toxicology, 163: 112929. . Gray, N. E., Harris, C. J., Quinn, J. F., and Soumyanath, A. (2016). Centella asiatica modulates antioxidant and mitochondrial pathways and improves cognitive function in mice. Journal of ethnopharmacology, 180, 78-86.

[0150] 3. Hao-Nan, W., Zheng, S., Qing, L., Xiao- Ying, H., Yan, C., Da-Hui, L., Hong, J., and Hong-

[0151] Zhi, D. (2020). Isochlorogenic acid (ICGA): natural medicine with potentials in pharmaceutical developments. Chinese journal of natural medicines, 18(11), 860-871. . Knafo, S., Sanchez-Puelles, C., Palomer, E., Delgado, I., Draffin, J. E., Mingo, J., Wahle,

[0152] T., Kaleka, K., Mou, L., Pereda-Perez, I., Klosi, E., Faber, E.B., Chapman, H.M., Lozano-Montes, L., Ortega-Molina, A., Ordonez-Gutierrez, L., Wandosell, F., Vina, J., Doth, C.G., Hall, R.A., Pilido, R., Gerges, N.Z., Chan, A.M., Spaller, M.R., Serrano, M., Venero, C., and Esteban, J. A. (2016). PTEN recruitment controls synaptic and cognitive function in Alzheimer's models. Nature neuroscience, 19(3), 443-453.

[0153] 5. Moerke, N. J. (2009). Fluorescence polarization (FP) assays for monitoring peptide-protein or nucleic acid-protein binding. Current protocols in chemical biology, 1(1), 1-15.

[0154] 6. Morris, R. G. (1981). Spatial localization does not require the presence of local cues.

[0155] Learning and motivation, 12(2), 239-260.

[0156] 7. Matthews, D. G., Caruso, M., Alcazar Magana, A., Wright, K. M., Maier, C. S., Stevens, J.

[0157] F., Gray, N.E., Quinn, J.F., and Soumyanath, A. (2020). Caffeoylquinic acids in Centella asiatica reverse cognitive deficits in male 5XFAD Alzheimer’s disease model mice. Nutrients, 12(11), 3488.

[0158] 8. Oakley, H., Cole, S. L., Logan, S., Maus, E., Shao, P., Craft, J., Guillozet-Bongaarts, A.,

[0159] Ohno, M., Disterhoft, J., Van Eldik, L., Berry R., and Vassar, R. (2006). Intraneuronal P-amyloid aggregates, neurodegeneration, and neuron loss in transgenic mice with five familial Alzheimer's disease mutations: potential factors in amyloid plaque formation. Journal of Neuroscience, 26(40), 10129-10140.

Claims

CLAIMSWe claim:

1. A method for inhibiting an interaction of phosphatase and tensin homologue deleted from chromosome 10 (PTEN) and PSD95 / Disc large / Zonula occludens-1 (PDZ), comprising administering to a subject in need thereof a composition comprising an effective amount of at least two compounds selected from amaranth, pentagalloylglucose, isochlorogenic acid C, isochl orogenic acid B, and hematoporphyrin dihydrochloride.

2. The method of claim 1, wherein the composition comprises isochlorogenic acid B and isochlorogenic acid C; amaranth and isochlorogenic acid C; isochlorogenic acid B and amaranth; or isochlorogenic acid B, isochlorogenic acid C, and amaranth.

3. The method of claim 1, wherein the subject has a neurodegenerative disorder.

4. The method of claim 3, wherein the neurodegenerative disorder is Alzheimer’s Disease.

5. The method of claim 1, wherein the administering comprises injecting the composition into the subject.

6. The method of claim 5, wherein the injecting is by intravenous, intramuscular, intraperitoneal, or subcutaneous injection.

7. The method of claim 1, wherein the administering comprises oral ingestion.

8. The method of claim 1, wherein the administering comprises nasal inhalation.

9. The method of claim 1, wherein the isochlorogenic acid B, isochlorogenic acidC, and / or amaranth are administered in an amount of about 5 mg / kg to about 100 mg / kg subject body weight.

10. The method of claim 9, wherein the isochl orogenic acid B, isochl orogenic acid C, and / or amaranth are administered in an amount of about 10 mg / kg to about 20 mg / kg subject body weight.

11. The method of claim 10, wherein the isochlorogenic acid B and / or isochlorogenic acid C are administered in an amount of about 20 mg / kg subject body weight.

12. The method of claim 10, wherein the amaranth is administered in an amount of about 10 mg / kg subject body weight.

13. The method of claim 1, wherein the pentagalloylglucose and / or hematoporphyrin dihydrochloride are administered in an amount of about 10 mg / kg subject body weight.

14. The method of claim 11, wherein the isochlorogenic acid B and / or isochlorogenic acid C are administered in an amount of about 20 mg / kg subject body weight and the amaranth is administered in an amount of about 10 mg / kg.

15. A composition comprising at least two compounds selected from amaranth, pentagalloylglucose, isochlorogenic acid C, isochlorogenic acid B, and hematoporphyrin dihydrochloride; and a pharmaceutically effective carrier and / or excipient.

16. The composition of claim 15, further comprising a therapeutic agent effective for treating a neurodegenerative disorder.

17. A kit comprising a first container containing a pharmaceutical composition comprising at least two compounds selected from amaranth, pentagalloylglucose, isochlorogenic acid C, isochlorogenic acid B, and hematoporphyrin dihydrochloride, and a second container containing a therapeutic agent effective for treating a neurodegenerative disorder.

18. The kit of claim 17, wherein the neurodegenerative disorder is Alzheimer's Disease.

19. The kit of claim 17, further comprising instructions for administration of the contents of the first container and the contents of the second container to a subject.