WO2013086327A1 - Nutritional compositions comprising curcumin and phosphatidylserine-docosahexaenoic acid for improving cognition - Google Patents

Abstract

Disclosed are nutritional compositions including a combination of Curcumin and/or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid. The combination improves cognitive performance in an individual, and, in one embodiment, improves a cognitive impairment and/or brain dysfunction associated with age-related cognitive decline or cognitive decline resulting from a neurodegenerative disease. This combination is also effective in improving memory acquisition, memory retention and recall in an individual.

Description

NUTRITIONAL COMPOSITIONS COMPRISING CURCUMIN AND

PHOSPHATIDYLSERINE-DOCOSAHEXAENOIC ACID FOR IMPROVING

COGNITION

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 61/568,338, filed on December 8, 2011, the contents of which are incorporated by reference herein.

FIELD OF THE DISCLOSURE

[0002] The present disclosure relates to nutritional compositions comprising Curcumin and phosphatidylserine-docosahexaenoic acid (PS-DHA) and to the methods of administering the compositions for improving cognitive performance and/or memory, enhancing memory acquisition and memory retention and recall in an individual, as well as reducing brain dysfunction and cognitive decline. In some embodiments, bioavailable Curcumin is utilized in combination with the PS-DHA.

BACKGROUND OF THE DISCLOSURE

[0003] Manufactured nutritional liquids and powders comprising a targeted selection of nutrition ingredients are well known and widely available, some of which may provide a sole source of nutrition while others may provide a supplemental source. These nutritionals include powders that can be reconstituted with water or other aqueous liquid, as well as ready to drink nutritional liquids such as milk or protein based emulsions or non-emulsified liquids. These nutritional liquids are especially useful when formulated with selected nutritional ingredients. [0004] An estimated 4 to 5 million Americans, about 15% of those older than age 65, have some form and degree of cognitive failure. Cognitive failure, which includes dysfunction or loss of cognitive function (e.g., learning, thinking and memory), commonly occurs in association with central nervous system (CNS) disorders or conditions, including neurodegenerative diseases. Cognitive dysfunction may cause significant impairment of social and/or occupational functioning, which can interfere with the ability of an individual to perform activities of daily living and greatly impact the autonomy and quality of life of the individual.

[0005] Accordingly, there has recently been an increased interest in designing and marketing so-called "smart formulations" that include nutritional products designed specifically for brain health and nourishment. Many of these products are specifically designed for improving cognition and preventing

neurodegenerative diseases and related cognitive decline. To date, these formulations and products have had limited success.

[0006] As such, there is a need for nutritional compositions and methods for enhancing cognitive performance, and particularly, memory acquisition and memory recall that may contribute to the learning and memory processes. It would be beneficial if such nutritional compositions and methods could also improve a cognitive impairment and/or brain dysfunction, such as from age-related cognitive decline or from neurodegenerative diseases.

SUMMARY OF THE DISCLOSURE

[0007] One embodiment is directed to a method for improving memory acquisition and memory retention and recall in an individual. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0008] Another embodiment is directed to a method for improving cognitive performance in an individual. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid. [0009] Another embodiment is directed to a method for improving memory acquisition in an individual. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0010] Another embodiment is directed to a method for improving memory retention and memory recall in an individual. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0011] Another embodiment is directed to a method for enhancing N- Methyl-D-aspartate receptor (NMDAR) dependent hippocampal long-term

potentiation (LTP), a physiological correlate or a measure of synaptic plasticity, which may be a key underlying molecular mechanism of learning and memory. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine- docosahexaenoic acid.

[0012] Another embodiment is directed to a method for enhancing the NMDAR and 2-amino-3-(5-methyl-3-oxo-l,2-oxazol-4-yl)propanoic acid receptor (AMPAR) mediated hippocampal synaptic plasticity, which may be another key mechanism underlying learning and memory. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0013] Another embodiment is directed to a method for preventing the over- activation of NMDAR, which may result in neuronal death. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0014] Another embodiment is directed to a method for enhancing neuronal membrane fluidity. The method comprises administering to the individual a composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid. [0015] Another embodiment is directed to a nutritional composition comprising an effective amount of Curcumin or bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

[0016] Nutritional compositions including Curcumin, and in particular bioavailable Curcumin, and PS-DHA in combination provide enhanced bioavailability of the individual components of Curcumin, phosphatidylserine (PS), and DHA to an individual upon administration. Therefore, this combination of components is more efficacious in providing improved cognitive functioning and/or performance to an individual, as well as reducing cognitive impairment and/or brain dysfunction in an individual. Specifically, the combination of Curcumin or bioavailable Curcumin and PS-DHA produces an added benefit on cognitive functioning by producing a synergistic effect in enhancing NMDAR and AMPAR dependent hippocampal synaptic plasticity, which is a molecular mechanism involved in learning and memory. In addition, Curcumin or bioavailable Curcumin and PS-DHA produces a sequential action by improving memory acquisition that is essential for the process of learning, and by improving memory retention and memory recall that is necessary for the retrieval of learned processes.

[0017] The combination of Curcumin or bioavailable Curcumin and PS- DHA may also provide benefits to an individual by its complementary mode of action on neuroprotection. Neuronal death due to the oxidative stress resulting from

NMDAR over-activation (i.e., NMDAR mediated excitotoxicity) is a major problem associated with cognitive impairment, brain dysfunction, and neurodegenerative diseases (e.g., Alzheimer's Disease). It has now been found that Curcumin, and particularly bioavailable Curcumin, offers significant neuroprotection by preventing NMDAR mediated excitotoxicity and resulting neuronal death.

[0018] Additionally, PS-DHA offers benefits such as the maintenance of neuronal membrane fluidity, which is essential for synaptic receptor function, and neurotransmitter release, which is essential for the cognitive performance of an individual. While PS-DHA may be taken up by the brain from the diet to provide the above benefits, PS-DHA is highly susceptible to oxidation and may therefore induce oxidative stress in neuronal membranes that may, in turn, negatively affect the neuronal health and function. It has now been found, however, that administration of Curcumin, and in particular bioavailable Curcumin, which possesses strong antioxidant properties, can prevent the oxidation of PS-DHA as well as provide neuroprotection and prevention of neuronal death due to oxidative stress, which is induced by increased calcium influx, thereby allowing the subsequent release of proteolytic enzymes, transcription factors, and reactive oxygen species. Therefore, Curcumin or bioavailable Curcumin and PS-DHA will have a complimentary mode of action to provide cognitive benefits.

[0019] Curcumin, and in particular bioavailable Curcumin, and PS-DHA act in combination to further provide anti-inflammatory and anti-amyloidogenic properties to an individual. Neuronal inflammation and amyloid β protein deposition are associated with cognitive impairment and/or brain dysfunction that results from age-related cognitive decline and/or cognitive decline associated with

neurodegenerative diseases. These anti-inflammatory and anti-amyloidogenic properties of Curcumin or bioavailable Curcumin and PS-DHA, therefore, provide added benefits when used in combination, thereby reducing cognitive impairment and/or brain dysfunction.

[0020] Accordingly, the nutritional compositions and methods of the present disclosure offer an alternative therapeutic or nutritional intervention option that may contribute to the improvement of cognitive performance, as well as to the reduction of cognitive impairment and brain dysfunction, in individuals, and particularly in adults, older adults, and the elderly.

BRIEF DESCRIPTION OF THE FIGURES

[0021] FIG. 1A is a chart depicting the effect of PS-DHA on memory acquisition, memory retention and memory recall as analyzed in Example 21.

[0022] FIG. IB is a chart depicting the effect of Curcumin on memory acquisition, memory retention and memory recall as analyzed in Example 21. [0023] FIG. 2A is a chart depicting the effect of Curcumin on NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0024] FIGS. 2B-2D are graphs depicting the effect of Curcumin on

NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0025] FIG. 3 A is a chart depicting the effect of PS-DHA on NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0026] FIGS. 3B-3D are graphs depicting the effect of PS-DHA on NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0027] FIG. 4A is a chart depicting the synergistic effect of the combination of Curcumin and PS-DHA on NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0028] FIGS. 4B-4D are graphs depicting the synergistic effect of the combination of Curcumin and PS-DHA on NMDAR dependent hippocampal LTP as analyzed in Example 22.

[0029] FIG. 5A is a chart depicting the neuroprotective effect of Curcumin on primary hippocampal neurons from NMDAR mediated excitotoxicity.

[0030] FIG. 5B is a chart depicting the neuroprotective effect of PS-DHA on primary hippocampal neurons from NMDAR mediated excitotoxicity.

DETAILED DESCRIPTION OF THE DISCLOSURE

[0031] The compositions and methods herein are directed to nutritional compositions comprising Curcumin, and in particular bioavailable Curcumin, and PS- DHA that can improve general cognitive performance in an individual, including memory acquisition, memory retention and memory recall. These and other essential or optional elements or features of the various embodiments are described in detail hereafter. [0032] The term "older adult" as used herein, unless otherwise specified, refers to an individual of at least 45 years of age, including at least 50 years of age, including at least 55 years of age, including at least 60 years of age, including at least 65 years of age, including at least 70 years of age, including at least 75 years of age, including at least 80 years of age, further including from about 55 years of age to about 80 years of age.

[0033] The term "Curcumin" refers to Curcumin and derivatives and analogs thereof.

[0034] The term "bioavailable" as used herein, unless otherwise specified, refers to the ability of a compound to enter into and remain in the bloodstream of an individual such that the substance can be absorbed into cells in the body. As the degree of bioavailability of a compound increases, the compound becomes more likely to enter into and remain in the bloodstream where it can be absorbed and used by the body. As the degree of bioavailability of a compound decreases, the compound becomes more likely to go directly into the gastrointestinal area and be expelled from the body before entering the bloodstream.

[0035] The term "nutritional composition" as used herein, unless otherwise specified, refers to nutritional liquids and nutritional powders that comprise at least one of protein, fat, and carbohydrate and is suitable for oral administration to a human. The nutritional composition may further comprise vitamins, minerals, and other ingredients and represent a sole, primary, or supplemental source of nutrition.

[0036] The terms "fat," "lipid" and "oil" as used herein, unless otherwise specified, are used interchangeably to refer to lipid materials derived or processed from plants or animals. These terms also include synthetic lipid materials so long as such synthetic materials are suitable for oral administration to humans.

[0037] The term "susceptible" as used herein, unless otherwise specified, means having little resistance to a certain condition or disease, including being genetically predisposed, having a family history of, and/or having symptoms of the condition or disease. [0038] The term "synergy" or "synergistic amount" as used herein, unless otherwise specified, refers to the interaction of two or more compounds so that their combined effect is greater than the additive sum of their individual effects.

[0039] The term "cognitive performance" as used herein, unless otherwise specified, refers to the learning, thinking, and memory functions (i.e., memory acquisition, memory retention and memory recall) of the brain. Accordingly, the term "improving cognitive performance" as used herein, unless otherwise specified, refers to improving the learning, thinking, and/or memory (memory acquisition, memory retention and memory recall) functions of an individual.

[0040] The term "improving a cognitive impairment and/or brain dysfunction" as used herein, unless otherwise specified, refers to the treating, preventing, and/or reducing the incidence or severity of cognitive decline associated with age-related cognitive decline or neurodegenerative disease.

[0041] The term "age-related cognitive decline" as used herein, unless otherwise specified, refers to a gradual decline in learning, thinking, and/or memory functions that are normal consequences of aging.

[0042] The term "neurodegenerative disease" as used herein, unless otherwise specified, refers to the progressive loss of structure or function of neurons, including the death of neurons and includes diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, dementia, amyotrophic lateral sclerosis, stroke, and schizophrenia.

[0043] All percentages, parts and ratios as used herein, are by weight of the total product, unless otherwise specified. All such weights as they pertain to listed ingredients are based on the active level and, therefore, do not include solvents or byproducts that may be included in commercially available materials, unless otherwise specified.

[0044] All references to singular characteristics or limitations of the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made.

[0045] All combinations of method or process steps as used herein can be performed in any order, unless otherwise specified or clearly implied to the contrary by the context in which the referenced combination is made.

[0046] The various embodiments of the nutritional compositions of the present disclosure may also be substantially free of any optional or selected essential ingredient or feature described herein, provided that the remaining composition or powder still contains all of the required ingredients or features as described herein. In this context, and unless otherwise specified, the term "substantially free" means that the selected composition contains less than a functional amount of the optional ingredient, typically less than about 1%, including less than about 0.5%, including less than about 0.1%, and also including zero percent, by weight of such optional or selected essential ingredient.

[0047] The nutritional compositions may comprise, consist of, or consist essentially of the essential elements of the products as described herein, as well as any additional or optional element described herein or otherwise useful in nutritional product applications.

Product Form

[0048] The nutritional compositions of the present disclosure may be formulated and administered in any known or otherwise suitable oral product form. Any solid, liquid, or powder form, including combinations or variations thereof, are suitable for use herein, provided that such forms allow for safe and effective oral delivery to the individual of the essential ingredients as also defined herein.

[0049] The nutritional compositions are most suitably formed as aqueous emulsions, including water-in-oil emulsions, oil-in-water emulsions, or complex (e.g., oil-in- water-in-oil emulsions) or other emulsion systems. As applied to the nutritional compositions herein, the nutritional emulsion embodiments are most typically oil-in- water emulsions comprising an internal or discontinuous oil phase that comprises the Curcumin or bioavailable Curcumin and PS-DHA as defined herein.

Curcumin and Bioavailable Curcumin

[0050] Curcumin, 1,7-bis (4-hydroxyphenyl)-l,6-heptadiene-3,5-dione and having the formula

is the principal Curcuminoid of turmeric. Conventionally, Curcumin has suffered lower bioavailability when taken orally, and thus when formulated at higher concentrations to counter its inherent poor bioavailability to achieve the desired systemic delivery, the products often take on an intense undesirable yellow color. In some embodiments, the present nutritional compositions use "bioavailable

Curcumin", which has an improved bioavailability as compared to conventionally used Curcumin. As such, the bioavailable Curcumin can be utilized in lower concentrations in the nutritional compositions and methods of the present disclosure, while still maintaining its anti-inflammatory, antioxidative, and anti-amyloidogenic activity.

[0051] The term "bioavailable Curcumin" refers to Curcumin and derivatives and analogs thereof, including natural and synthetic derivatives of Curcumin, as well as any combination of one or more of Curcumin and a derivative and/or analog. In particular, the term "bioavailable Curcumin" should be understood to encompass compounds having a 1,7-bis (4-hydroxyphenyl)-l,6-heptadiene-3,5- dione or l,7-bis(4-hydroxyphenyl) hept-4-en-3-one skeleton wherein the phenyl groups independently may bear one or more alkoxy residues, especially one methoxy residue in the 3-position. In some embodiments, additional Curcuminoids, such as demethoxyCurcumin and bisdemethoxyCurcumin, may also be present in the nutritional compositions. When present, demethoxyCurcumin and

bisdemethoxyCurcumin may be present as part of a complex with Curcumin. [0052] The "bioavailable Curcumin" used in the nutritional compositions of the present disclosure shows improved oral bioavailability as compared to Curcumins that are not substantially "bioavailable." The oral bioavailability can be determined in experiments involving oral administration of the bioavailable Curcumin composition of the present disclosure (and/or a corresponding amount of non-bioavailable

Curcumin) to a subject and measuring the level of the Curcumin in a biological sample obtained from the subject over time, wherein the biological sample may be derived from a body fluid, for example serum, plasma, whole blood, or cerebrospinal fluid, and/or a tissue, e.g. from brain, liver, kidney, or heart. For analysis, the Curcumin level in the examined body fluid or tissue may be plotted against time, and the area under the curve (AUC), for example the total area under the curve from t = 0 (time of administration) to t = infinity (= AUCO-infinity), or the area under the curve within a defined period, e.g. from t = 0 to t = 6 hours (AUC0-6H), may be calculated. In general, a higher AUC relative to the AUC obtained by administration of non- bioavailable Curcumin indicates an improved bioavailability. The absolute bioavailability may be calculated from the resulting AUC data as percentage based on the corresponding AUC data obtained from intravenous administration of Curcumins.

[0053] In some embodiments, the bioavailable Curcumin amount in the blood, determined as AUC0-6H after a single oral administration of a dose of the bioavailable Curcumin-containing nutritional composition of the present disclosure corresponding to 20 mg of total Curcumin to a human subject or an animal subject, preferably a rat, is significantly higher than after oral administration of the same amount of non-bioavailable Curcumin in the composition, preferably at least 2 times, at least 3 times, at least 4 times, at least 6 times, at least 8 times, at least 10 times, or at least 15 times, and, for example, up to 30 times higher.

[0054] As used herein, the amount of Curcumin or bioavailable Curcumin in the blood being "significantly higher" means a statistically significant increase of this parameter in subjects after oral administration of 20 mg of bioavailable Curcumin in the nutritional composition of the present disclosure as compared to the control 20 mg of Curcumin that is not bioavailable. A statistical test known in the art, such as ANOVA or Student's t-test, may be used to determine the significance of this difference, wherein the p-value is at least <0.1, <0.5, <0.01, <0.005, <0.001 or O.OOOl .

[0055] Bioavailable Curcumin can be prepared in a number of ways including, for example, using Meltrex® or similar melt-extrusion technology to prepare extruded solids and improve the bioavailability of the Curcumin as compared to Curcumin not produced by melt extrusion. Meltrex® or similar melt-extrusion technology methods are known in the art and can be applied to produce bioavailable Curcumin by one skilled in the art based on the disclosure herein.

[0056] In another embodiment, Curcumin can be co-supplemented with piperine (generally extracted from black pepper) to increase the bioavailability and hence the absorbability of Curcumin. In one embodiment, the piperine is co- supplemented in an amount of about 20 mg to increase the bioavailability of the Curcumin.

[0057] In another embodiment, the Curcumin or bioavailable Curcumin may be solubilized in an oil having an HLB of from about 0.7 to about 14 (polar oils) such that the resulting oil mixture provides increased bioavailability of Curcumin. One suitable polar oil for dissolving the Curcumin is a medium chain triglyceride oil (MCT oil).

[0058] In one embodiment of the present disclosure, the bioavailable Curcumin is a mixture of Curcuminoids (i.e., Curcumin, demethoxyCurcumin and bisdemethoxyCurcumin) obtained from the rhizomes of Curcuma Longa. In another embodiment, the bioavailable Curcumin is obtained using Meltrex® technology (Abbott Nutrition, Columbus, Ohio). In yet another embodiment, the bioavailable Curcumin is Meriva Bioavailable Curcumin, commercially available from Idena SPA (Milan, Italy).

[0059] The Curcumin or bioavailable Curcumin concentration in the nutritional compositions may range from at least about 0.001%, including from about 0.002%) to about 3.4%, including from about 0.002%> to about 3.36%, including from about 0.005%) to about 1.87%, also including from about 0.03%> to about 0.935%), also including from about 0.1% to about 0.5%, also including from about 0.1 % to about 0.467%), and also including from about 0.234%> to about 0.3%>, by weight of the nutritional composition. Exemplary embodiments of the present disclosure include nutritional compositions having Curcumin or bioavailable Curcumin in amounts ranging from 0.002% to about 0.234%, from about 0.005% to about 0.467%, from about 0.03% to about 0.935%, from about 0.1% to about 1.87%, and from about 0.3% to about 3.36%, by weight of the nutritional composition.

[0060] The nutritional compositions of the present disclosure desirably include sufficient Curcumin or bioavailable Curcumin to provide an individual with at least about 1 milligram, including at least about 3 milligrams, including from about 10 milligrams to about 10,000 milligrams, including from about 100 milligrams to about 4000 milligrams, including from about 400 milligrams to about 2000 milligrams, including from about 1200 milligrams to about 1800 milligrams, per day of Curcumin or bioavailable Curcumin. The total daily amount of Curcumin or bioavailable Curcumin may be administered to an individual in a single undivided dose, or may be split into one, two, three, four or more doses per day.

Phosphatidylserine-docosahexaenoic acid (PS-DHA)

[0061] In addition to the Curcumin, and in some embodiments, bioavailable Curcumin, the nutritional compositions of the present disclosure include PS-DHA. PS-DHA is a conjugated form of PS to DHA or a DHA enriched form of PS, with palmitic acid at the Sn-1 position and DHA at the Sn-2 position. The inclusion of PS- DHA in the nutritional compositions provide enhanced bioavailability of both PS and DHA in the brain as conjugated DHA allows for a more efficient uptake of DHA by the brain as compared to DHA in triglyceride form.

[0062] Generally, PS is a phospholipid component constituting 10%> of the total phospholipids present in the human body and 15% of PS is found in the brain. DHA is an omega-3 polyunsaturated fatty acid (PUFA) and is abundant in the brain and retina, accounting for 40% of the PUFAs in the brain and 60% of the PUFAs in the retina. DHA is essential for the proper functioning of adult brains, deficiencies of which have been associated with cognitive decline.

[0063] PS-DHA is the natural form of PS. Common sources of PS-DHA include marine, plant and animal sources such as mackerel, chicken, herring, eel, pig, tuna, clams, veal, beef, pork, turkey, cod, anchovy, and trout and/or plant sources such as soybeans, cabbage, whole-grain barley, rice, and carrots.

[0064] The PS-DHA concentration in the nutritional compositions may range from at least about 0.001%, including from about 0.001% to about 1.2%, including from about 0.001% to about 1.121%, including from about 0.01% to about 0.6%), including from about 0.02%> to about 0.2%>, also including from about 0.02%> to about 0.1%), also including from about 0.025%) to about 0.09%>, and also including from about 0.025% to about 0.04%, by weight of the nutritional composition.

Exemplary embodiments of the present disclosure include nutritional compositions having PS-DHA in amounts ranging from 0.001% to about 0.037%, from about 0.002% to about 0.094%, from about 0.02% to about 0.187%, from about 0.025% to about 0.561%, and from about 0.04% to about 1.121%, by weight of the nutritional composition.

[0065] The nutritional compositions of the present disclosure desirably include sufficient PS-DHA to provide an individual with from about 100 milligrams to about 2000 milligrams, including from about 200 milligrams to about 2000 milligrams, including from about 200 milligrams to about 1000 milligrams, including from about 500 milligrams to about 1500 milligrams, including from about 1000 milligrams to about 1400 milligrams, per day of PS-DHA. The total daily amount of PS-DHA may be administered to an individual in a single undivided dose, or may be split into one, two, three, four or more doses per day.

Macronutrients

[0066] The nutritional compositions of the present disclosure may further comprise one or more optional macronutrients in addition to the Curcumin or bioavailable Curcumin and PS-DHA described herein. The optional macronutrients include proteins, lipids, carbohydrates, and combinations thereof. The nutritional compositions are desirably formulated as dietary products containing all three macronutrients.

[0067] Macronutrients suitable for use herein include any protein, lipid, or carbohydrate or source thereof that is known for or otherwise suitable for use in an oral nutritional composition, provided that the optional macronutrient is safe and effective for oral administration and is otherwise compatible with the other ingredients in the nutritional composition.

[0068] The concentration or amount of optional lipid, carbohydrate, and protein in the nutritional composition can vary considerably depending upon the particular product form (e.g., bars or other solid dosage forms, milk or soy-based liquids or other clear beverages, reconstitutable powders, etc.) and the various other formulations and targeted dietary needs. These optional macronutrients are most typically formulated within any of the embodied ranges described in the following tables.

Each numerical value preceded by the term "about"

Each numerical value preceded by the term "about"

Carbohydrate [0069] Optional carbohydrates suitable for use in the nutritional

compositions may be simple, complex, or variations or combinations thereof, all of which are optionally in addition to the Curcumin or bioavailable Curcumin as described herein. Non-limiting examples of suitable carbohydrates include hydrolyzed or modified starch or cornstarch, maltodextrin, isomaltulose, sucromalt, glucose polymers, sucrose, corn syrup, corn syrup solids, rice-derived carbohydrate, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), and combinations thereof.

[0070] Optional carbohydrates suitable for use herein also include soluble dietary fiber, non-limiting examples of which include gum Arabic,

fructooligosaccharide (FOS), sodium carboxymethyl cellulose, guar gum, citrus pectin, low and high methoxy pectin, oat and barley glucans, carrageenan, psyllium and combinations thereof. Insoluble dietary fiber is also suitable as a carbohydrate source herein, non-limiting examples of which include oat hull fiber, pea hull fiber, soy hull fiber, soy cotyledon fiber, sugar beet fiber, cellulose, corn bran, and combinations thereof.

Protein

[0071] Optional proteins suitable for use in the nutritional compositions include hydrolyzed, partially hydrolyzed or non-hydro lyzed proteins or protein sources, and can be derived from any known or otherwise suitable source such as milk (e.g., casein, whey), animal (e.g., meat, fish, egg albumen), cereal (e.g., rice, corn), vegetable (e.g., soy, pea, potato), or combinations thereof. The proteins for use herein can also include, or be entirely or partially replaced by, free amino acids known for use in nutritional products, non-limiting examples of which include L-tryptophan, L- glutamine, L-tyrosine, L-methionine, L-cysteine, taurine, L-arginine, L-carnitine, and combinations thereof.

Lipid

[0072] Optional lipids suitable for use in the nutritional compositions, which are optionally in addition to the PS-DHA as described herein, include coconut oil, fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil, high oleic safflower oil, high GLA-safflower oil, MCT oil (medium chain triglycerides), sunflower oil, high oleic sunflower oil, palm and palm kernel oils, palm olein, canola oil, flaxseed oil, borage oil, cottonseed oils, evening primrose oil, blackcurrant seed oil, transgenic oil sources, fungal oils, marine oils (e.g., tuna, sardine) and so forth.

Optional Ingredients

[0073] The nutritional compositions may further comprise other optional ingredients that may modify the physical, nutritional, chemical, hedonic or processing characteristics of the products or serve as pharmaceutical or additional nutritional components when used in a targeted population. Many such optional ingredients are known or otherwise suitable for use in other nutritional products and may also be used in the nutritional compositions described herein, provided that such optional ingredients are safe and effective for oral administration and are compatible with the essential and other ingredients in the composition.

[0074] Non-limiting examples of such other optional ingredients include preservatives, anti-oxidants, buffers, pharmaceutical actives, sweeteners, colorants, flavors, flavor enhancers, thickening agents and stabilizers, emulsifying agents, lubricants, and combinations thereof.

[0075] The nutritional compositions may further include one or more minerals, non-limiting examples of which include phosphorus, sodium, chloride, magnesium, manganese, iron, copper, zinc, iodine, calcium, potassium, chromium, molybdenum, selenium, and combinations thereof.

[0076] The nutritional compositions may also include one or more vitamins, non-limiting examples of which include carotenoids (e.g., beta-carotene, zeaxanthin, lutein, lycopene), biotin, choline, inositol, folic acid, pantothenic acid, TP AN, choline, vitamin A, thiamine (vitamin Bl), riboflavin (vitamin B2) niacin (vitamin B3), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), ascorbic acid (vitamin C), vitamin D, vitamin E, vitamin K, and various salts, esters, or other derivatives thereof, and combinations thereof. Methods of Manufacture

[0077] The nutritional compositions may be prepared by any known or otherwise effective manufacturing technique for preparing the selected product form. Many such techniques are known for any given product form such as nutritional liquids and nutritional powders and can easily be applied by one of ordinary skill in the nutrition and formulation arts to the nutritional products described herein.

[0078] Liquid, milk or soy-based nutritional liquids, for example, may be prepared by first forming an oil and fiber blend containing all formulation oils, any emulsifier, fiber and fat-soluble vitamins. Additional slurries (typically a

carbohydrate and two protein slurries) are prepared separately by mixing the carbohydrate and minerals together and the protein in water. The slurries are then mixed together with the oil blend. The resulting mixture is homogenized, heat processed, standardized with any water-soluble vitamins, flavored and the liquid terminally sterilized or aseptically filled or dried to produce a powder.

[0079] The compositions of the present disclosure may also be manufactured by other known or otherwise suitable techniques not specifically described herein without departing from the spirit and scope of the present disclosure. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive and that all changes and equivalents also come within the description of the present disclosure.

Methods of Use

[0080] The methods include the oral administration of the nutritional compositions that include Curcumin or bioavailable Curcumin in combination with PS-DHA, to improve cognitive performance in any individual. Particularly, the combination of bioavailable Curcumin and PS-DHA may improve general cognition by producing a sequential action on memory acquisition, memory retention and memory recall that contributes to the cognitive functions of learning, thinking, and memory. [0081 ] In addition to memory acquisition and memory recall, the nutritional compositions including the combination of Curcumin or bioavailable Curcumin and PS-DHA may have a complementary mode of action in maintaining neuronal membrane fluidity, allowing for proper hormone and neurotransmitter function. This may further improve cognitive performance.

[0082] Additionally, in some embodiments, the nutritional compositions can be utilized to improve a cognitive impairment and/or brain dysfunction that may be associated with a neurodegenerative disease. Particularly, the combination of Curcumin or bioavailable Curcumin and PS-DHA shows anti-amyloido genie properties and anti-inflammation properties, thereby reducing neuronal inflammation and clearance of amyloid β protein deposit that can lead to a cognitive impairment and/or brain dysfunction associated with neurodegenerative diseases or conditions such as Alzheimer's disease, Huntington's disease, Parkinson's disease, dementia, amyotrophic lateral sclerosis, stroke, and/or schizophrenia. By reducing this neuronal inflammation and clearing of the amyloid protein, cognitive function may be improved.

[0083] Furthermore, in some embodiments, the nutritional compositions of the present disclosure may improve a cognitive impairment and/or brain dysfunction associated with age-related cognitive decline or cognitive decline associated with a neurodegenerative disease by enhancing synaptic plasticity, which is controlled by N- methyl-D-aspartate receptor (NMDAR). Deficits in synaptic plasticity and/or neuronal death can result in cognitive impairment and/or brain dysfunction. The composition restores the suboptimum level of NMDAR activity to optimum levels, even during weak stimulation, and, at the same time, prevents neuronal death from NMDAR over-activation. Consequently, NMDAR dependent hippocampal long term potentiation (LTP) is enhanced, which is one key underlying molecular mechanism of learning and memory. Further, by preventing neuronal death, the neuronal loss associated with cognitive impairment and brain dysfunction can be avoided. In another embodiment, the nutritional compositions may improve learning and memory in an individual by enhancing the NMDAR and 2-amino-3-(5-methyl-3-oxo-l,2- oxazol-4-yl) propanoic acid receptor (AMPAR) mediated hippocampal synoptic plasticity.

[0084] Although in some embodiments the methods of the present disclosure may be directed to individuals who have a neurodegenerative disease or condition, or a disease or condition related to a neurodegenerative disease or condition, the methods of the present disclosure as described herein are also intended in some embodiments to include the use of such methods in "at risk" individuals, including individuals unaffected by or not otherwise afflicted with neurodegenerative diseases or conditions such as those described above, for the purpose of preventing, minimizing, or delaying the development of such diseases or conditions over time. For such prevention purposes, the methods of the present disclosure preferably include continuous, daily administration of the compositions as described herein. Such preventive methods may be directed at adults or others, particularly older adults, who are susceptible to developing neurodegenerative diseases due to hereditary considerations, environmental considerations, and the like.

[0085] The individual desirably consumes at least one serving of the nutritional composition daily, and in some embodiments, may consume two, three, or even more servings per day. Each serving is desirably administered as a single, undivided dose, although the serving may also be divided into two or more partial or divided servings to be taken at two or more times during the day. The methods of the present disclosure include continuous day after day administration, as well as periodic or limited administration, although continuous day after day administration is generally desirable. The methods of the present disclosure are preferably applied on a daily basis, wherein the daily administration is maintained continuously for at least 3 days, including at least 5 days, including at least 1 month, including at least 6 weeks, including at least 8 weeks, including at least 2 months, including at least 6 months, desirably for at least about 18-24 months, desirably as a long term, continuous, daily, dietary supplement.

EXAMPLES [0086] The following examples illustrate specific embodiments and or features of the nutritional products of the present disclosure. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present disclosure, as many variations thereof are possible without departing from the spirit and scope of the disclosure.

[0087] The exemplified products are nutritional products prepared in accordance with manufacturing methods well known in the nutrition industry for preparing nutritional liquids (e.g., emulsions and clear liquids) and powders.

Examples 1-5

[0088] Examples 1-5 illustrate nutritional emulsions of the present disclosure, the ingredients of which are listed in the table below. All ingredient amounts are listed as kg per 1000 kg batch of product, unless otherwise specified.

Table 1: Nutritional Emulsions

Examples 6-10

[0089] Examples 6-10 illustrate nutritional emulsions of the present disclosure, the ingredients of which are listed in the table below. All ingredient amounts are listed as kg per 1000 kg batch of product, unless otherwise specified. Table 2: Nutritional Emulsions

Examples 11-15

[0090] Examples 11-15 illustrate nutritional clear liquids of the present disclosure, the ingredients of which are listed in the table below. All ingredient amounts are listed as kg per 1000 kg batch of product, unless otherwise specified. Table 3: Clear Liquids

EXAMPLES 16-20

[0091] Examples 16-20 illustrate nutritional powders of the present disclosure, the ingredients of which are listed in the table below. These products are prepared by spray drying methods in separate batches, and are reconstituted with water prior to use to the desired target ingredient concentrations. All ingredient amounts are listed as kg per 1000 kg batch of product, unless otherwise specified.

Table 4: Nutritional Powders

Sodium phosphate monobasic 1.6 1.6 1.6 1.6 1.6 monohydrate

Potassium phosphate dibasic trihydrate 1.1 1.1 1.1 1.1 1.1

Art. Cotton candy flavor 1.0 1.0 1.0 1.0 1.0

Vitamin premix 1.0 1.0 1.0 1.0 1.0

Ascorbyl palmitate 0.243 0.243 0.243 0.243 0.243

Ascorbic acid 0.240 0.240 0.240 0.240 0.240

Antioxidant 0.116 0.116 0.116 0.116 0.116

Ferrous sulfate 0.010 0.090 0.030 0.020 0.070

Vitamin premix 0.065 0.065 0.065 0.065 0.065

Zinc sulfate monohydrate 0.057 0.057 0.057 0.057 0.057

Manganese sulfate 0.045 0.045 0.045 0.045 0.045

Mineral mix copper sulfate 0.035 0.035 0.035 0.035 0.035

Beta carotene 30% 0.005 0.005 0.005 0.005 0.005

Chromium chloride 0.001 0.001 0.001 0.001 0.001

Sodium molybdate 0.0012 0.0012 0.0012 0.0012 0.0012

Potassium iodide 0.001 0.001 0.001 0.001 0.001

Sodium selenite 0.0004 0.0004 0.0004 0.0004 0.0004

Citric acid AN AN AN AN AN

Potassium hydroxide AN AN AN AN AN

Magnesium sulfate dry AN AN AN AN AN

Ultra micronized tricalcium phosphate AN AN AN AN AN

Ascorbic acid AN AN AN AN AN

AN = As Needed

EXAMPLE 21

[0092] In this Example, the efficacy of Curcumin and PS-DHA in enhancing cognitive performance by improving hippocampal function in normal adult rats was analyzed.

[0093] Hippocampal dysfunction due to altered NMD A and AMP A

mediated synaptic plasticity is known to cause spatial memory deficits and cognitive decline. Hippocampus is involved in spatial memory both in humans and rodents. In this Example, the efficacy of Curcumin and PS-DHA separately on enhancing

cognitive performance by improving hippocampal function at various concentrations is investigated in normal adult rats using Morris water maze task, which is a well known animal behavioral paradigm to test the hippocampal based spatial memory in rodents.

[0094] Male albino Wistar rats (Harlan Sprague-Dawley, Inc., USA), 2-3 months old, were individually housed in polycarbonate cages with Bed-O-Cob® bedding in a temperature-controlled room (25°C) with a 12-hour light/dark cycle. The rats (i.e., test subjects) were handled beginning the day after arrival, allowed free access to water, and provided with a control diet (standard rodent diet AIN 93 G, available from Harlan, USA) or with test diets (AIN 93 G with respective amounts of PS-DHA added) shown below for two weeks prior to water maze testing and throughout the testing period.

[0095] Water Maze Test: To determine the effects of the test diets on spatial reference learning, water maze tests were performed in a circular pool (diameter: 180 cm, height: 76 cm) made of black plastic. The pool was filled to a depth of 35 cm of water (maintained at 25.0 ± 1.0°C). The pool was located in a large room with a number of extra-maze visual cues including geometric images (e.g., squares, triangles, circles etc.) hung on the wall, ambient lighting, approximately 25-30 Lux (lumen/m ), and black curtains, used to hide the experimenter (visually) and the resting test subjects. Swimming activity of each rat was monitored via a video camera mounted overhead, which relayed information including latency to find the platform, total distance traveled, time and distance spent in each quadrant etc. to a video tracking system (Noldus Etho Vision 3.1).

[0096] Hidden Platform Task: For these experiments, an invisible (black) 10 cm x 10 cm square platform was submerged approximately 1.0 cm below the surface of the water and placed in the center of the northeast quadrant (which remained constant throughout hidden platform training). Each rat was given one trial per day for 14 consecutive days to locate and climb on to the hidden platform. A trial was initiated by placing the rat in the water directly facing the pool wall (i.e., nose approximately 2 cm from the wall) in one of the 4 quadrants. The daily order of entry into individual quadrants was pseudo-randomized such that all 4 quadrants were used once every four training days. For each trial, the rat was allowed to swim a maximum of 90 seconds in order to find the platform. When successful, the rat was allowed a 30-second rest period on the platform. If unsuccessful within the allotted time period, the rat was given a score of 90 seconds and then physically placed on the platform and also allowed the 30-second rest period. In either case the rat was subsequently returned to its home cage. Latency in seconds (A) and distance swam in centimeters (B) to locate a hidden platform over 14 consecutive days (sessions) corresponded to the measure of memory acquisition. Effect of PS-DHA in enhancing memory acquisition is depicted in FIG. 1 A.

[0097] Probe Trials (Transfer Tests): Twenty-four hours following the last hidden platform trial (i.e., day 15 of testing), a probe trial was conducted in which the platform was removed from the pool to measure spatial bias and the recall memory of the previous platform location. This was accomplished by measuring the percentage of time spent in the previous target quadrant and the number of crossings over of the previous platform location. The time spent in target quadrant corresponded to an estimate of the strength of the recall memory of the previous platform location. The number of crossings over of the previous platform location was correlated to the strength of memory retention. Effect of Curcumin in enhancing the recall memory is shown in FIG. IB.

[0098] Visible Platform Task: After probe trial tests on day 15, a visible platform test was performed in order to assure that the study subjects were visually capable of performing the task and that they demonstrated normal search/escape behaviors. To accomplish this task, a highly visible (white) cover fitted with a small white flag was attached to the platform (dimensions with cover attached = 12 cm x 12 cm), which raised the surface approximately 1.0 cm above the surface of the water. Each rat was gently lowered into the water in the quadrant diametrically opposite to the platform quadrant and given one or more trials with a 90-second time limit to locate and climb onto the platform. If unsuccessful after 90 seconds, it was physically placed on the platform for 30 seconds then given a new trial. Once a rat was successful on its own accord, it was then given a series of 4 additional trials (with a 1.0-minute intertrial interval) and the latency (in seconds) to locate the platform was recorded. The platform was moved on each trial to a different quadrant (the subject was always entered from the opposite quadrant) until the test was conducted once in all 4 quadrants.

[0099] As shown in FIG. 1 A, in normal adult rats, PS-DHA administration at 90mg/Kg body weight had a significant lowering effect on latency in seconds (A) and distance swam in centimeters (B) to locate a hidden platform, and therefore PS- DHA had a significant enhancing effect on memory acquisition.

[00100] As shown in FIG. IB, in normal adult rats, Curcumin administration at 100 mg/Kg body weight significantly increased the percentage of time spent in the previous target quadrant that corresponded to an estimate of the strength of the recall memory. Similarly, an increasing trend in the number of platform crossings were also observed in rats administered with 100 or 300 mg/Kg body weight of Curcumin, indicating an improvement in memory retention. These findings suggest that a combination of Curcumin and PS-DHA holds the potential to produce a sequential action on memory acquisition, memory retention and memory recall that contributes to the processes of learning and memory.

EXAMPLE 22

[0101] In this Example, the administration of Curcumin and PS-DHA, alone and in combination, on NMDAR dependent hippocampal LTP was analyzed.

[0102] Long lasting changes in the strength of AMPA and NMDA receptor mediated glutamatergic synaptic transmission in the hippocampus, known as hippocampal synaptic plasticity, is a key underlying molecular mechanism in learning and memory. AMPA and NMDA dependant hippocampal synaptic plasticity is associated with trafficking and delivery of AMPA receptors at the synapse in response to NMDAR activation. Previous studies have shown that in cognitive decline associated with aging or Alzheimer's disease onset, the strength of AMPA and NMD A synaptic transmission is compromised due to the internalization of NMD A and AMPA receptors, leading to deficits in NMDAR and AMPAR expression at the synapse. During learning, neuronal activation leads to pre-synaptic release of glutamate resulting in the activation of post-synaptic AMPA and NMD A receptors. Activation of post-synaptic AMPA receptors results in the flow of Na⁺ ions, causing the depolarization of post-synaptic neuron. This depolarization abolishes the blocking

2__|_

of NMD A receptors by Mg , resulting in the opening of NMD A receptor channels for Na⁺ and Ca²⁺ ions. Ca²⁺ influx through NMD A receptors is known to trigger

2__|_

intracellular signaling cascades, notably the phosphorylation of Ca /calmodulin- dependent protein kinase (CaMKII) that phosphorylates AMPA receptors and modulates the channel properties, ultimately resulting in long-term potentiation (LTP), the physiological correlate of synaptic plasticity (learning and memory).

[0103] Therefore in this Example, NMDAR dependant hippocampal LTP was used as a physiological marker to assess the efficacy of Curcumin and PS-DHA in enhancing cognitive function. To evaluate the efficacy of Curcumin and PS-DHA in enhancing cognitive function, NMDAR dependant LTP was recorded on hippocampal brain slices in vitro.

[0104] Experiments were carried out with 7-9 weeks-old C57/Black6 mice (from Elevage Janvier, Le Genest St Isle, France). Animals were housed and used in accordance to the French and European legislations for animal care. The mice were sacrificed by fast decapitation, without previous anaesthesia. The brain was quickly removed and soaked in ice-cold oxygenated buffer with the following composition:

Cutting Solution Final concentration (niM)

D-glucose 11

saccharose

250

[0105] Hippocampus slices (350 μιη) were cut with a Macllwain tissue- chopper and incubated at room temperature for at least 60 minutes in Artificial Cerebro-Spinal Fluid (ACSF) having the following composition:

During experiments, slices were continuously perfused with oxygenated ACSF.

[0106] PS-DHA was prepared as a 60, 30 or 15 mM stock solution in a mix (10% Cremphor El (ref: C5135, Sigma, batch: 1439553 13509161) - 5% ethanol (ref: 20820.293, VWR, batch: 07K210530)-85% ACSF) from powder on each day of experimentation. This stock solution was then 1000X diluted into ACSF to reach the final 60, 30 or 15 μΜ concentration.

[0107] Curcumin was prepared as a 30, 15 or 5 mM Stock solution in DMSO, this solution was then 10X diluted in ACSF containing 1 eq. NaOH (i.e. 300, 150 or 50 μΜ NaOH, ref: 28 244.295, VWR, batch: 0601507). This solution was finally 100X diluted in ACSF to reach the final 30, 15 or 5 μΜ concentration.

[0108] D-AP5 (ref: Asc-271, ASCENT, batch: APN08163-1-1) was dissolved as a 30 mM stock solution in water, aliquoted and stored at -20°C until use. Aliquots were thawed and vortexed each day of experiment and then 1000X or 10000X diluted in ACSF to reach the final concentration of 30 μΜ or 3 μΜ. [0109] NBQX (ref: Asc-045, ASCENT, batch: APN07044-8-3) was dissolved as a 10 mM stock solution in water, aliquoted and stored at -20°C until use. Aliquots were thawed and vortexed each day of experiment and then 1000X diluted in ACSF to reach the final concentration of 10 μΜ.

[0110] All data was recorded with a MEA set-up, commercially available from Multichannel Systems MCS GmbH (Reutlingen, Germany), and composed of a 4-channel stimulus generator and a 60-channels amplifier head-stage connected to a 60-channels A/D card. Software for stimulation, recordings and analysis were commercially available from Multi Channel Systems: MC Stim (3.2.4 release) and MC Rack (4.0.0 release), respectively. All of the experiments were carried out with 3-dimensional MEA (Ayanda Biosystems, S.A., CH-1015 Lausanne, Switzerland) that consisted of 60 tip-shaped and 60^m-high electrodes spaced by 100 μιη. The MEA electrodes were made of platinum with kΩ 450 < impedance < 600 kΩ.

[0111] A 350-μιη thick mouse hippocampal slice was disposed on the multi- electrode array (100 μιη distant electrodes). One electrode was chosen to stimulate Schaeffer collaterals at the CA3/CA1 interface. An I/O curve was performed to monitor evoked-responses for stimulations between 100 and 800 μΑ, by 100 μΑ steps. The stimulus was a monopolar biphasic current pulse (negative for 60 μβ and then positive for 60 μβ), settled to evoke 40% of the maximal amplitude response (as determined with the I/O curve) and applied every 30 seconds to evoke "responses" (i.e. field Excitatory Post Synaptic Potentials; fEPSP) in the CA1 region.

[0112] Short term memory formation, accompanied by a weak potentiation, effect of Curcumun and PS-DHA on LTP induced by a weak tetanus was also analyzed. Following a 10-minute period to verify the baseline stability of fEPSP to elicite a weak potentiation, a weak tetanus (10 stimuli at 100 Hz for 0.1 s at 20% of IMAX (Kanno et al, Brain research, 2004)) was used. Weak tetanus-induced potentiation was followed over a 40-minute period.

[0113] Evoked-responses (fEPSP) were recorded if they satisfied quality criteria described in Standard Operating Procedures: correct location, stable baseline (fluctuation within +/- 10 % during ten consecutive minutes), amplitude > 100 μν after background noise subtraction. The fEPSP from selected electrodes were simultaneously sampled at 5 kHz and recorded on the hard disk of a PC until offline analysis. In parallel, fEPSP amplitudes of selected electrodes were compiled online (with MC Rack program) to monitor and to follow the good performance of the experiments. Data was plotted in a standard spreadsheet file for off-line analysis. The fEPSP resulted from glutamatergic synaptic transmission consecutive to afferent pathway stimulation. At the end of each experiment, 10 μΜ NBQX were perfused on the slice to validate the glutamatergic nature of synaptic transmission as well as to subtract background noise at individual electrode level. Control LTP were recorded in parallel, with hippocampal slices prepared from the same animals as the ones used to evaluate the compounds.

[0114] During experiments, the slices were continuously perfused with ACSF solutions (bubbled with 95% 0₂-5% C0₂) at the rate of 3 mL/min with a peristaltic pump (ME A chamber volume: ~1 mL). Complete solution exchange in the ME A chamber was achieved 20 seconds after the switch of solutions. The perfusion liquid was continuously pre-heated at 37°C just before reaching the MEA chamber with a heated-perfusion cannula (PH01, Multichannel Systems, Reutlingen,

Germany). The temperature of the MEA chamber was maintained at 37 +/- 0.1 °C with a Peltier element located in the MEA amplifier headstage.

[0115] I/O curves from identical experimental conditions were analysed using repeated measure two-way ANOVA followed by a post hoc Bonferroni test. LTP from identical experimental conditions were analysed using one-way ANOVA followed by a post hoc Dunnett test. Statistical analysis was performed by using Prism 5.0 software. A critical P value P<0.05 was considered significant for the statistical tests used throughout the study.

[0116] As shown in the table below and in FIGS. 2A-2D, Curcumin at 2.5, 5, and 10 μΜ significantly enhanced NMDAR dependant hippocampal LTP induced by weak tetanic frequency stimulation (one trains of 100 Hz stimulations for 0.1 seconds). As more particularly shown in FIG. 2C, this significant enhancement in NMDAR dependent hippocampal LTP was more pronounced at 5 μΜ. Moreover, Curcumin was found to be effective in enhancing NMDAR dependent hippocampal LTP at lower ranges of concentrations. For example, Curcumin significantly reduced LTP at 15 μΜ.

[0117] As shown in FIGS. 3A and 3C-3D, PS-DHA at 5 and 10 μΜ significantly enhanced NMDAR dependant hippocampal LTP induced by weak tetanic frequency stimulation (one trains of 100 Hz stimulations for 0.1 seconds) of Schaffer collaterals of CA1 region of hippocampus. The effect of PS-DHA at 2.5 μΜ on NMDAR dependent hippocampal LTP induced by weak titanic frequency stimulation is also shown in FIG. 3B and in the table below. Weak stimulation resulted in weak potentiation that normally occurs during short term memory formation.

[0118] As shown in FIGS . 4A-4D (particularly, FIG. 4D, and in the tables below, a combination of Curcumin at 2.5 μΜ and PS-DHA at 2.5 μΜ produced a synergistic effect in enhancing NMDAR dependant hippocampal LTP induced by weak tetanic frequency stimulation (one trains of 100 Hz stimulations for 0.1 seconds) of Schaffer collaterals of CA1 region of hippocampus. Weak stimulation resulted in weak potentiation that normally occurs during short term memory formation.

[0119] Accordingly, Curcumin and PS-DHA showed significant enhancement of hippocampal LTP by modulating the NMDAR activity. In addition, Curcumin and PS-DHA showed a synergistic effect in enhancing hippocampal LTP by modulating the NMDAR activity.

EXAMPLE 23

[0120] In this Example, the ability of Curcumin and PS-DHA to provide neuroprotection to hippocampal neurons from NMDAR mediated excitotoxicity was analyzed.

[0121] One of the major molecular events that contribute to neuronal death or damage in brain malfunctioning, cognitive impairment and neurodegenerative diseases, including Alzheimer's disease (AD), is glutamate or NMDAR mediated excitotoxicity. Glutamate is the major excitatory neurotransmitter in the brain that activates the major ion channel receptors such as NMDAR and AMPAR that contribute to synaptic plasticity. However, excessive release of glutamate and overstimulation of NMDAR can cause neuronal death or damage due to increased calcium influx resulting in the release of proteolytic enzymes, transcription factors and reactive oxygen species. Therefore, compounds or ingredients that prevent NMDAR mediated excitotoxicity will offer neuroprotection. NMDA is a specific agonist of NMDAR that is capable of causing the excitotoxic neuronal death due to

2__|_

the over activation of NMDAR in Mg free medium. The NMDA receptor channel is highly permeable to Ca 2+ but is blocked by Mg 2+ in a voltage-dependent manner. Therefore, in this Example, to induce NMDAR mediated excitotoxicity, NMDA was applied in the presence of different concentrations of Curcumin and PS-DHA in a

2__|_

Mg free medium to hippocampal neurons. Neuroprotective effects of Curcumin and PS-DHA from NMDAR mediated excitotoxicity were evaluated from the amount of lactate dehydrogenase (LDH) enzyme, a stable cytoplasmic enzyme that is released when the plasma membrane is damaged and released into the cell-culture supernatant in response to NMDAR mediated excitotoxicity.

[0122] Hippocampal neurons were plated in 96-well plates and cultured for 10 days. NMDA and glutamate neurotoxicity assays were performed as detailed below. All preincubation experiments were performed in neurobasal medium

2__|_

containing Mg , which was omitted only when NMDA was added for 15 minutes in Hanks' balanced salt solution containing 10 μΜ glycine and 2.7 mM CaCl₂. After 10 days in culture, hippocampal neurons were incubated for 15 minutes with Curcumin (0, 1, 5.0, 10, 20 and 40 μΜ) and PS-DHA (10, 30 and 90 μΜ) in the absence or presence of 200 μΜ NMDA. Cell viability was determined using cytotoxicity assay kits (LDH release, Roche) following the manufacturers' instructions.

[0123] The results demonstrated that Curcumin showed a significant inhibition of LDH release caused by NMDA induced cell death at 5, 10 and 20 μΜ (n=4, p<0.05, FIG. 5A). Curcumin at 1 μΜ showed a significant increase in LDH release caused by NMDA induced cell death. In contrast, PS-DHA showed significant inhibition of LDH release caused by NMDA induced cell death (i.e., offered neuroprotection) only at 10 μΜ (FIG. 5B). [0124] Accordingly, Curcumin showed a comparatively significant neuroprotection by inhibiting the over activation of NMDAR mediated ex cito toxicity as compared to PS-DHA (FIGS. 5 A and 5B). Curcumin offers significant neuroprotection by preventing NMDAR mediated excitotoxicity and resulting neuronal death due to the oxidative stress induced by the increased calcium influx and subsequent release of proteolytic enzymes, transcription factors and reactive oxygen species.

Claims

WHAT IS CLAIMED IS:

1. A method for improving at least one of cognitive performance, memory acquisition, memory recall, memory acquisition and memory recall, and combinations thereof in an individual, the method comprising administering to the individual a composition comprising an effective amount of bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

2. The method of claim 1 comprising from about 0.002% to about 3.4% bioavailable Curcumin by weight of the composition and from about 0.001% to about 1.2% phosphatidylserine-docosahexaenoic acid by weight of the composition.

3. The method of claim 1 comprising from about 0.1% to about 0.5% bioavailable Curcumin by weight of the composition and from about 0.02% to about 0.1%) phosphatidylserine-docosahexaenoic acid by weight of the composition.

4. The method of claim 1 wherein the individual is administered from about 400 milligrams/day to about 2000 milligrams/day of bioavailable Curcumin and from about 100 milligrams/day to about 2000 milligrams/day of phosphatidylserine- docosahexaenoic acid.

5. The method of claim 1 wherein the individual has age-related cognitive decline.

6. The method of claim 1 wherein the individual has a neurodegenerative disease selected from the group consisting of Alzheimer's disease, Huntington's disease, Parkinson's disease, dementia, amyotrophic lateral sclerosis, stroke, and schizophrenia.

7. The method of any one of claims 1-6, wherein the method improves cognitive performance in the individual.

8. The method of any one of claims 1-6, wherein the method improves memory acquisition in the individual.

9. The method of any one of claims 1-6, wherein the method improves memory recall in the individual.

10. The method of any one of claims 1-6, wherein the method improves memory acquisition and memory recall in the individual.

11. The nutritional composition of any one of claims 1-4.

12. A nutritional composition comprising an effective amount of bioavailable Curcumin and phosphatidylserine-docosahexaenoic acid.

13. The nutritional composition of claim 12 comprising from about 0.002% to about 3.4% bioavailable Curcumin by weight of the composition and from about 0.001%) to about 1.2% phosphatidylserine-docosahexaenoic acid by weight of the composition.

14. The nutritional composition of claim 12 comprising from about 0.1% to about 0.5% bioavailable Curcumin by weight of the composition and from about 0.02%) to about 0.1 % phosphatidylserine-docosahexaenoic acid by weight of the composition.

15. The nutritional composition of any one of claims 12-14 further comprising at least one macronutrient selected from the group consisting of proteins, lipids, carbohydrates, sources thereof, and combinations thereof.