Use of lacto-n-difucohexaose-i to enhance brain performance

Abstract

The invention relates to the use of lacto-N-difucohexaose-I or a synthetic composition comprising thereof for contributing to brain health of a human.

Description

[0001] USE OF LACTO-N-DIFUCOHEXAOSE-I TO ENHANCE BRAIN PERFORMANCE

[0002] FIELD OF THE INVENTION

[0003] The invention relates to the use of lacto-N-difucohexaose-l or a synthetic composition comprising thereof for contributing to brain health of a human.

[0004] BACKGROUND OF THE INVENTION

[0005] Lacto-N-difucohexaose-l (LNDFH-I, Fuca1-2Gaipi-3[Fuca1-4]GlcNAcpi-3Gaipi-4Glc) is a human milk oligosaccharide (HMO) exhibiting prominent physiological characteristics, including modifying the intestinal microbiota, immunomodulation, anti-bacterial activities and antiviral infection (Yang et al. Crit. Rev. Food Sci. Nutr. https: / / doi.org / 10.1080 / 10408398.2023.2224431 (2023)). Although it is the second most abundant HMO after 2’-fucosyllactose (2’-FL) in terms of g / l concentration (Soyyilmaz et al. Nutrients 13, 2737 (2021) in human milk, it has received lesser attention than 2’-FL.

[0006] N-acetyl-L-glutamine (NAG) is a central nervous system stimulant which can directly pass through the blood-brain barrier (Zhang et al. J. Pharm. Anal. 7, 303 (2017)). NAG is considered as glutamine (Gin) substitution and becoming indispensable in some diseases and injury treatment. Although Gin serves as a direct building block for protein synthesis and supports the tricarboxylic acid cycle, it can undergo degradation into ammonia which induces cell toxicity. Previous studies indicate that NAG seems to have a similar pharmacological function to Gin, while it does not produce toxic degradation product in solution. NAG, through its conversion to glutamine, serves as a crucial precursor for the synthesis of glutamate, the primary excitatory neurotransmitter in the brain. Glutamate is essential for synaptic transmission, learning, and memory. Glutamine is also a precursor for gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter. By contributing to the production of both excitatory and inhibitory neurotransmitters (glutamate and GABA), NAG helps maintain the balance necessary for optimal cognitive function, including learning and memory. Proper neurotransmitter balance is essential for synaptic plasticity, which underlies learning and long-term memory formation. During brain development, glutamine is involved in the proliferation and differentiation of neural progenitor cells. This process is essential for the formation of a functional neural network. Glutamine is vital for the production of adenosine triphosphate (ATP), the primary energy currency of cells. In the brain, adequate levels of glutamine support mitochondrial function and energy production, crucial for maintaining cognitive processes and overall brain health. By supporting neurotransmitter synthesis and energy production, NAG can enhance cognitive functions such as attention, memory, and learning. Glutamine contributes to the synthesis of glutathione, one of the brain’s most important antioxidants. Glutathione protects neurons from oxidative stress and free radical damage and therefore provides neuroprotective effect.

[0007] N-acetyl-L-tyrosine (NAT) is a derivative of the amino acid tyrosine, which is involved in several critical functions in the brain. Tyrosine itself is a non-essential amino acid that serves as a precursor for the synthesis of important neurotransmitters such as dopamine, norepinephrine, and epinephrine. The acetylation of tyrosine enhances its solubility and stability, potentially improving its bioavailability and absorption. By supporting the production of neurotransmitters, NAT helps maintain healthy brain function. By boosting the levels of dopamine and norepinephrine, NAT can enhance cognitive functions such as attention, focus, and memory. These neurotransmitters are involved in executive functions and working memory (Fernstrom et al. J. Nutr. 137, 1539S (2007)).

[0008] Nicotinic acid, also known as niacin or vitamin B3, is an essential nutrient involved in numerous metabolic processes, including energy production and DNA repair. It can be synthesized in the gut from the amino acid tryptophan by gut microbiota. The ability of nicotinic acid synthesized in the gut to reach the brain involves several steps, including absorption into the bloodstream, crossing the blood-brain barrier (BBB), and utilization by brain cells. Nicotinic acid crosses the BBB via facilitated diffusion or active transport mechanisms involving specific carrier proteins. These transport systems ensure that adequate levels of nicotinic acid reach the brain. This process is crucial for maintaining adequate levels of NAD+, which is essential for energy production, DNA repair, and overall neuronal health. NAD+ is involved in the metabolism of several neurotransmitters. Proper neurotransmitter levels are crucial for synaptic transmission, mood regulation, cognition, and overall brain function. Nicotinic acid and its derivatives can contribute to the synthesis of glutathione, one of the brain's primary antioxidants. Glutathione helps protect neurons from oxidative stress and free radical damage.

[0009] There is a need to find further, advantageously more beneficial methods, to enhance the production of metabolites contributing to brain health.

[0010] SUMMARY OF THE INVENTION

[0011] In a first aspect, this invention provides LNDFH-I for use in: the preservation or protection of neurons and neural tissue, enhancing the production of neurotransmitters, supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient being in the state or at risk of developing a neurological disorder.

[0012] In a second aspect, this invention provides a synthetic composition comprising LNDFH-I for use in: the preservation or protection of neurons and neural tissue, enhancing the production of neurotransmitters, supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient being in the state or at risk of developing a neurological disorder.

[0013] The synthetic composition can be a nutritional or pharmaceutical composition. Advantageously, the synthetic composition is not mammalian milk or processed mammalian milk.

[0014] A third aspect of this invention is a method for the preservation or protection of neurons and neural tissue of a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl- L-glutamine, and nicotinic acid.

[0015] A fourth aspect of this invention is a method for enhancing the production of neurotransmitters in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0016] A fifth aspect of this invention is a method for supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0017] A sixth aspect of this invention is a method for supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0018] A seventh aspect of the invention is a method for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a human, the method comprising orally or enterally administering to or providing for consumption by the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I.

[0019] An eighth aspect of the invention is a method for enhancing brain performance and perception of well-being, the method comprising orally or enterally administering to or providing for consumption by the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N- acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0020] A ninth aspect of the invention relates to a non-therapeutic use of LNDFH-I for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a healthy human. In one embodiment, LNDFH-I is comprised in a nutritional composition which is preferably not mammalian milk or processed mammalian milk.

[0021] A tenth aspect of the invention is a non-therapeutic use of LNDFH-I a) to enhance brain performance and perception of well-being in a healthy human, or b) for protection of neurons and neural tissue in a healthy human, advantageously to enhance brain performance and perception of well-being. In one embodiment, LNDFH-I is comprised in a nutritional composition which is preferably not mammalian milk or processed mammalian milk.

[0022] BRIEF DESCRIPTION OF THE FIGURES

[0023] The invention will be described in further detail hereinafter with reference to the accompanying figures, in which:

[0024] Figure 1 shows the AUG values of metabolites produced in the experiment disclosed in Example 1.

[0025] DETAILED DESCRIPTION OF THE INVENTION

[0026] “About” means + / - 5 %.

[0027] The phrase "biological process that governs the growth, differentiation, and maturation of the nervous system" refers to the complex set of mechanisms and events that control the development and functioning of the nervous system. This process begins during embryonic development and continues throughout life, involving a series of stages that are tightly regulated by genetic and environmental factors. It is critical for normal development and functioning of the brain and spinal cord. The growth phase of the nervous system involves the proliferation and migration of neural cells, which form the basic structure of the nervous system. This is followed by the differentiation phase, during which neural cells become specialized and acquire distinct functions, such as sensory perception, motor control, and cognitive processing. Finally, the maturation phase involves the refinement and optimization of neural connections, leading to the formation of complex neural networks that underlie behaviour and cognition.

[0028] “Cognitive development” refers to the progressive growth and development of a person's thinking, reasoning, problem-solving, and decision-making abilities. This process starts from infancy and continues throughout adulthood.

[0029] “Cognitive function” refers to the mental processes and abilities that enable an individual to carry out tasks related to thinking, understanding, learning and remembering. It includes various aspects such as mental clarity, memory, focus, problem-solving, decision-making and reasoning. These functions are essential for everyday activities and overall mental performance.

[0030] “Dietary management" means exclusive or partial feeding of subjects who, because of a disease, disorder or medical condition: either have a limited, impaired or disturbed capacity to take, digest, absorb, metabolise or excrete ordinary food or certain nutrients contained therein, or metabolites, or have other medically-determined nutrient requirements

[0031] (see: Commission Notice on the classification of Food for Special Medical Purposes of the European Commission, Official Journal of the European Union C 401, 25.11.2017, p. 10-11).

[0032] "Effective amount" means an amount of a composition that provides an HMO in a sufficient amount to render a desired treatment outcome in a human. An effective amount can be administered or provided for consumption in one or more doses to achieve the desired treatment outcome.

[0033] “Enteral administration” means any conventional form for delivery of a composition to a human that causes the deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include feeding through a naso-gastric tube or jejunum tube, oral, sublingual and rectal.

[0034] “Healthy” means enjoying health, that is the condition of an organism in which it performs its vital functions normally or properly; when free from physical disease or pain and not likely to develop such. “Human” can be an infant (younger than 3 years of age) or a non-infant (a child, a teenager, an adult or an elderly person).

[0035] “Language skills” refers to the ability to understand and use the language. Language development begins in infancy and continues through childhood and adolescence. Language development involves several key milestones, including learning to recognize and differentiate between speech sounds, developing a vocabulary of words, learning grammar and syntax, and developing the ability to use language for communication.

[0036] “Learning” can be defined as the process of acquiring new knowledge, skills, behaviours, or attitudes through experience, study, or instruction. It involves a change in behaviour or mental processes that results from experience or practice. Learning can occur through a variety of methods, including observation, trial and error, feedback, and instruction. The ability to learn is developed early in life and continues to develop throughout the lifespan. Infants are born with the ability to learn and are constantly learning about their environment through their senses and experiences. As they grow, their ability to learn becomes more sophisticated and complex, allowing them to acquire new knowledge and skills.

[0037] “Motor actions” or “motor skills” relate to the coordination of various muscles and body parts to achieve a specific goal. Examples of motor skills include walking, running, jumping, throwing, and catching.

[0038] “Neurological disorder” is a disorder of the nervous system including the brain, spinal cord and peripheral nerves, resulting in various functional impairments. A neurological disorder can lead to symptoms such as cognitive deficits, motor dysfunction, sensory disturbances and emotional changes.

[0039] “Neurotransmitter” is a signalling molecule secreted by a neuron to affect another cell across a synapse.

[0040] “Non-therapeutical use” means - as opposed to the purpose of therapy which is to restore the organism from a pathological to its original condition, or to prevent pathology in the first place - an improvement of performance of the organism in a normal state. Therefore, the purpose of the non-therapeutical use is to enhance performance and perception of well-being in a healthy mammal / individual.

[0041] "Oral administration" means any conventional form for the delivery of a composition to a human through the mouth. Accordingly, oral administration is a form of enteral administration.

[0042] The phrase "preservation and protection of neurons and neural tissue" refers to the maintenance of the structure and function of nerve cells and the tissue that makes up the nervous system, preventing damage or degeneration of neurons and neural tissue, and promoting their survival and health. Some of the factors that can contribute to damage or degeneration of neurons and neural tissue include aging, injury, disease, and exposure to toxins or other harmful substances.

[0043] “Preventive treatment” or “prevention” means treatment given or action taken to diminish the risk of onset or recurrence of a disease.

[0044] “Spatial recognition” relates to the ability to perceive, analyse, and understand spatial relationships between objects and the environment. This includes the ability to recognize and remember the location of objects in space, to mentally manipulate objects in space, and to navigate through the environment.

[0045] “Synthetic composition” means a composition which is artificially prepared and preferably means a composition containing at least one compound that is produced ex vivo chemically and / or biologically, e.g. by means of chemical reaction, enzymatic reaction or recombinantly. The synthetic composition typically comprises an active component, here LNDFH-I, that is capable of enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid. In some embodiments, the synthetic composition may comprise one or more compounds or components other than HMOs that may have a beneficial effect on the microbiota of a human subject microbiota in vivo, e.g. non-digestible oligosaccharides or prebiotics. Also, in some embodiments, the synthetic compositions may comprise one or more nutritionally or pharmaceutically active components which do not affect adversely the efficacy of the above- mentioned compound. Some non-limiting embodiments of a synthetic composition of the invention are also described below.

[0046] “Therapy” means treatment given or action taken to reduce or eliminate symptoms of a disease or pathological condition.

[0047] “Treat” means to address a medical condition or disease with the objective of improving or stabilising an outcome in the person being treated or addressing an underlying nutritional need. Treat, therefore, includes the dietary or nutritional management of the medical condition or disease by addressing nutritional needs of the person being treated. “Treating” and “treatment” have grammatically corresponding meanings.

[0048] It has now been surprisingly found that LNDFH-I, in simulated colonic fermentation test, enhanced the production of metabolites, namely N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid, that contribute to the healthy brain functions. Even more surprisingly, the metabolite enhancing effect of LNDFH-I was more prominent than that of 2’-FL, the most abundant human milk oligosaccharide in mother’s milk and which has been growingly used to supplement conventional infant formulas or as dietary supplement for adults. Therefore, it can be concluded that LNDFH-I, by oral or enteral ingestion, dynamically modulates the human intestinal microbiota by preferentially promoting the production of the above mentioned metabolites and thus contributes to the healthy brain functions and perception of well-being, preferably more effectively than 2’-FL.

[0049] Accordingly, it is provided LNDFH-I for use in:

[0050] - the preservation or protection of neurons and neural tissue,

[0051] - enhancing the production of neurotransmitters,

[0052] - supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or

[0053] - supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient being in the state or at risk of developing a neurological disorder.

[0054] LNDFH-I may be used as neat, in other embodiment it may be used in blend with other HMOs.

[0055] In other aspect of the invention, it is provided a synthetic composition comprising LNDFH-I for use in

[0056] - the preservation or protection of neurons and neural tissue,

[0057] - enhancing the production of neurotransmitters,

[0058] - supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or

[0059] - supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient being in the state or at risk of developing a neurological disorder.

[0060] Another aspect of the invention relates to a non-therapeutic use of LNDFH-I for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a healthy human including human not at risk of developing disease such as a neurological disorder. In one embodiment, LNDFH-I is comprised in a synthetic composition, preferably in a nutritional composition which is preferably not mammalian milk or processed mammalian milk. By the non-therapeutic, when consuming LNDFH-I, use the healthy human receives no therapeutic benefit; however they receive benefit concerning performance improvement in general and a perception of well-being, especially in connection with brain functions. Preferably, the performance improvement and / or perception of well-being is more effective than the human would receive from 2’-FL consumption. In one embodiment, the production of N-acetyl-L-tyrosine and N-acetyl-L- glutamine is enhanced. In other embodiment, the production of N-acetyl-L-tyrosine and nicotinic acid is enhanced. In other embodiment, the production of N-acetyl-L-glutamine and nicotinic acid is enhanced. In other embodiment, the production of N-acetyl-L-tyrosine, N-acetyl-L- glutamine and nicotinic acid is enhanced.

[0061] A further aspect of the invention is a non-therapeutic use of LNDFH-I a) to enhance brain performance and perception of well-being in a healthy human including human not at risk of developing disease such as a neurological disorder, or b) for protection of neurons and neural tissue in a healthy human including human not at risk of developing disease such as a neurological disorder, advantageously to enhance brain performance and perception of wellbeing. In one embodiment, LNDFH-I is comprised in a synthetic composition, preferably in a nutritional composition which is preferably not mammalian milk or processed mammalian milk. Typically, to benefit from the non-therapeutical use, the human consumes LNDFH-I and experiences improvement e.g. in cognitive function, mental clarity, memory, focus. Preferably, by consumption of LNDFH-I the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a healthy human is enhanced that leads to enhanced brain performance and perception of well-being, for example by a better protection of neurons and neural tissue. Enhancing brain performance is often pursued by individuals who seek to optimize their cognitive abilities or function for personal or professional reasons, such as athletes, students, or professionals in demanding fields (with other word: headworkers) requiring work or effort consisting principally or largely of mental activity, thought, imagination, etc. (as opposed to physical or manual work). It may also be pursued by individuals who are experiencing cognitive decline or decline in cognitive development due to aging, typically elderly individuals above 60 years of age, and wish to improve their cognitive function. Further, healthy infants can also benefit from LNDFH-I consumption wherein the growth, differentiation and maturation of the nervous system is optimized contributing to perception well-being.

[0062] Preferably, the enhancement of brain performance is more effective compared to 2’-FL consumption. In one embodiment, the production of N-acetyl-L-tyrosine and N-acetyl-L- glutamine is enhanced. In other embodiment, the production of N-acetyl-L-tyrosine and nicotinic acid is enhanced. In other embodiment, the production of N-acetyl-L-glutamine and nicotinic acid is enhanced. In other embodiment, the production of N-acetyl-L-tyrosine, N-acetyl-L- glutamine and nicotinic acid is enhanced.

[0063] The synthetic composition, especially the synthetic composition comprising LNDFH-I for the use as disclose above, can be a pharmaceutical composition. The pharmaceutical composition can contain a pharmaceutically acceptable carrier, e.g. phosphate buffered saline solution, mixtures of ethanol in water, water and emulsions such as an oil / water or water / oil emulsion, as well as various wetting agents or excipients. The pharmaceutical composition can also contain other materials that do not produce an adverse, allergic or otherwise unwanted reaction when administered to humans. The carriers and other materials can include solvents, dispersants, coatings, absorption promoting agents, controlled release agents, and one or more inert excipients, such as starches, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrating agents. If desired, tablet dosages can be coated by standard aqueous or non-aqueous techniques.

[0064] The pharmaceutical compositions can be administered orally, e.g. as a tablet, capsule, or pellet containing a predetermined amount, or as a powder or granules containing a predetermined concentration or a gel, paste, solution, suspension, emulsion, syrup, bolus, electuary, or slurry, in an aqueous or non-aqueous liquid, containing a predetermined concentration. Orally administered compositions can include binders, lubricants, inert diluents, flavouring agents, and humectants. Orally administered compositions such as tablets can optionally be coated and can be formulated to provide sustained, delayed or controlled release of the mixture therein.

[0065] The pharmaceutical compositions can also be administered by rectal suppository, aerosol tube, naso-gastric tube or direct infusion into the Gl tract or stomach.

[0066] The pharmaceutical compositions can also include therapeutic agents such as antiviral agents, antibiotics, probiotics, analgesics, and anti-inflammatory agents. The proper dosage of these compositions for a human can be determined in a conventional manner, based upon factors such immune status, body weight and age. Also, the pharmaceutical composition can contain one or more human milk oligosaccharides other than LNDFH-I.

[0067] The synthetic composition, especially in the case of the non-therapeutical use of LNDFH-I as disclosed above, can be a nutritional composition. It can contain sources of protein, lipids and / or digestible carbohydrates and can be in powdered or liquid forms. The composition can be designed to be the sole source of nutrition or a nutritional supplement. The nutritional composition is preferably for non-therapeutic use.

[0068] Suitable protein sources include milk proteins, soy protein, rice protein, pea protein and oat protein, or mixtures thereof. Milk proteins can be in the form of milk protein concentrates, milk protein isolates, whey protein or casein, or mixtures of both. The protein can be whole protein or hydrolysed protein, either partially hydrolysed or extensively hydrolysed. Hydrolysed protein offers the advantage of easier digestion which can be important for humans with inflamed Gl tracts. The protein can also be provided in the form of free amino acids. The protein can comprise about 5 % to about 30 % of the energy of the nutritional composition, normally about 10 % to 20 %.

[0069] The protein source can be a source of glutamine, threonine, cysteine, serine, proline, or a combination of these amino acids. The glutamine source can be a glutamine dipeptide and / or a glutamine enriched protein. Glutamine can be included due to the use of glutamine by enterocytes as an energy source. Threonine, serine and proline are important amino acids for the production of mucin. Mucin coats the Gl tract and can improve mucosal healing. Cysteine is a major precursor of glutathione, which is key for the antioxidant defences of the body.

[0070] Suitable digestible carbohydrates include maltodextrin, hydrolysed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, high fructose corn syrup, rice-derived carbohydrates, pea-derived carbohydrates, potato-derived carbohydrates, tapioca, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), or mixtures thereof. Generally digestible carbohydrates provide about 35 % to about 55 % of the energy of the nutritional composition. Preferably the nutritional composition is free from lactose. A particularly suitable digestible carbohydrate is a low dextrose equivalent (DE) maltodextrin.

[0071] Suitable lipids include medium chain triglycerides (MCT) and long chain triglycerides (LCT).

[0072] Preferably the lipid is a mixture of MCTs and LCTs. For example, MCTs can comprise about 30 % to about 70 % by weight of the lipids, more specifically about 50 % to about 60 % by weight. MCTs offer the advantage of easier digestion which can be important for humans with inflamed Gl tracts. Generally, the lipids provide about 35 % to about 50 % of the energy of the nutritional composition. The lipids can contain essential fatty acids (omega-3 and omega-6 fatty acids).

[0073] Preferably these polyunsaturated fatty acids provide less than about 30 % of total energy of the lipid source. Decreasing the levels of these polyunsaturated fatty acids is believed to decrease sensitivity to peroxidation; which can be beneficial for humans having inflammatory conditions.

[0074] Suitable sources of long chain triglycerides are rapeseed oil, sunflower seed oil, palm oil, soy oil, milk fat, corn oil, high oleic oils, and soy lecithin. Fractionated coconut oils are a suitable source of medium chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a polyunsaturated fatty acid omega-6 (n-6) to omega-3 (n-3) ratio of about 4:1 to about 10:1. For example, the n-6 to n-3 fatty acid ratio can be about 6:1 to about 9:1.

[0075] The nutritional composition preferably also includes vitamins and minerals. If the nutritional composition is intended to be a sole source of nutrition, it preferably includes a complete vitamin and mineral profile. Examples of vitamins include vitamins A, B-complex (such as B1, B2, B6 and B12), C, D, E and K, niacin and acid vitamins such as pantothenic acid, folic acid and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium and boron.

[0076] The nutritional composition can also include a carotenoid such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoid included can vary from about 0.001 pg / ml to about 10 pg / ml. Lutein can be included in an amount of from about 0.001 pg / ml to about 10 pg / ml, preferably from about 0.044 pg / ml to about 5 g / ml of lutein. Lycopene can be included in an amount from about 0.001 pg / ml to about 10 pg / ml, preferably about 0.0185 mg / ml to about 5 g / ml of lycopene. Beta-carotene can comprise from about 0.001 pg / ml to about 10 mg / ml, for example about 0.034 pg / ml to about 5 pg / ml of beta-carotene.

[0077] The nutritional composition preferably also contains reduced concentrations of sodium; for example, from about 300 mg / l to about 400 mg / l. The remaining electrolytes can be present in concentrations set to meet needs without providing an undue renal solute burden on kidney function. For example, potassium is preferably present in a range of about 1180 to about 1300 mg / l; and chloride is preferably present in a range of about 680 to about 800 mg / l.

[0078] The nutritional composition can also contain various other conventional ingredients such as preservatives, emulsifying agents, thickening agents, buffers, fibres and prebiotics (e.g. fructooligosaccharides, galactooligosaccharides), one or more human milk oligosaccharides other than LNDFH-I, probiotics (e.g. B. animalis subsp. lactis BB-12, B. lactis HN019, B. lactis Bi07, B. infantis ATCC 15697, L. rhamnosus GG, L. rhamnosus HNOOI, L. acidophilus LA-5, L. acidophilus NCFM, L. fermentum CECT5716, B. longum BB536, B. longum AH1205, B. longum AH1206, B. breve M-16V, L. reuteri ATCC 55730, L. reuteri ATCC PTA-6485, L. reuteri DSM 17938), antioxidant / anti-inflammatory compounds including tocopherols, carotenoids, ascorbate / vitamin C, ascorbyl palmitate, polyphenols, glutathione, and superoxide dismutase (melon), other bioactive factors (e.g. growth hormones, cytokines, TFG-P), colorants, flavours, and stabilisers, lubricants, and so forth.

[0079] The nutritional composition can be in the form of a soluble powder, a liquid concentrate, or a ready-to-use formulation. The composition can be fed to a human via a nasogastric tube or orally. Various flavours, fibres and other additives can also be present.

[0080] The nutritional compositions can be prepared by any commonly used manufacturing techniques for preparing nutritional compositions in solid or liquid form. For example, the composition can be prepared by combining various feed solutions. A protein-in-fat feed solution can be prepared by heating and mixing the lipid source and then adding an emulsifier (e.g. lecithin), fat soluble vitamins, and at least a portion of the protein source while heating and stirring. A carbohydrate feed solution is then prepared by adding minerals, trace and ultra-trace minerals, thickening or suspending agents to water while heating and stirring. The resulting solution is held for 10 minutes with continued heat and agitation before adding carbohydrates (e.g. the HMOs and digestible carbohydrate sources). The resulting feed solutions are then blended together while heating and agitating and the pH adjusted to 6.6-7.0, after which the composition is subjected to high-temperature short-time processing during which the composition is heat treated, emulsified and homogenized, and then allowed to cool. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range, if necessary, flavours are added, and water is added to achieve the desired total solid level. For a liquid product, the resulting solution can then be aseptically packed to form an aseptically packaged nutritional composition. In this form, the nutritional composition can be in ready-to- feed or concentrated liquid form. Alternatively, the composition can be spray-dried and processed and packaged as a reconstitutable powder.

[0081] When the nutritional product is a ready-to-feed nutritional liquid, the total concentration of HMOs in the liquid, by weight of the liquid, is from about 0.0001 % to about 2.0 %, including from about 0.001 % to about 1.5 %, including from about 0.01 % to about 1.0 %. When the nutritional product is a concentrated nutritional liquid, the total concentration of HMOs in the liquid, by weight of the liquid, is from about 0.0002 % to about 4.0 %, including from about 0.002 % to about 3.0 %, including from about 0.02 % to about 2.0 %.

[0082] The nutritional composition can also be in a unit dosage form such as a capsule, tablet or sachet. For example, the synthetic composition can be in a tablet form comprising the HMOs, and one or more additional components to aid formulation and administration, such as diluents, excipients, antioxidants, lubricants, colorants, binders, disintegrants, and the like.

[0083] Suitable diluents, excipients, lubricants, colorants, binders, and disintegrants include polyethylene, polyvinyl chloride, ethyl cellulose, acrylate polymers and their copolymers, hydroxyethyl-cellulose, hydroxypropylmethyl-cellulose (HPMC), sodium carboxymethylcellulose, polyhydroxyethyl methylacrylate (PHEMA), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), polyethylene oxide (PEO), or polyacrylamide (PA), carrageenan, sodium alginate, polycarbophil, polyacrylic acid, tragacanth, methyl cellulose, pectin, natural gums, xanthan gum, guar gum, karaya gum, hypromellose, magnesium stearate, microcrystalline cellulose, and colloidal silicon dioxide. Suitable antioxidants are vitamin A, carotenoids, vitamin C, vitamin E, selenium, flavonoids, polyphenols, lycopene, lutein, lignan, coenzyme Q10 ("CoQIO") and glutathione.

[0084] The unit dosage forms, especially those in sachet form, can also include various nutrients including macronutrients.

[0085] The nutritional composition intended for infants is an infant or toddler formula.

[0086] For human administration, the dosage of LNDFH-I will be at a concentration similar to that found for the HMO in human breast milk. The required amount would generally be in the range from about 200 mg to about 20 g per day, in certain embodiments from about 300 mg to about 15 g per day, from about 400 mg to about 10 g per day, in certain embodiments from about 500 mg to about 10 g per day, in certain embodiments from about 1 g to about 10 g per day. Appropriate dose regimes can be determined by conventional methods.

[0087] Further aspects of the invention relate to a method for the preservation or protection of neurons and neural tissue, enhancing the production of neurotransmitters, supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient, preferably the human patient is in the state or at risk of developing a neurological disorder, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I as disclosed above. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0088] Furthermore, the invention relates to a method for enhancing brain performance and perception of well-being, the method comprising orally or enterally administering to or providing for consumption by the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I. Preferably, the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N- acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid. Preferably, the human is healthy and not at risk of developing disease such as a neurological disorder; such human receives benefit concerning performance improvement in general and a perception of well-being from the method, especially in connection with brain functions.

[0089] Also, the invention relates to a method for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a human, the method comprising orally or enterally administering to or providing for consumption by the human an effective amount of LNDFH-I, advantageously in a form of a synthetic composition comprising LNDFH-I as disclosed above. The above mentioned metabolites are believed to contribute to the healthy brain functions. The human may be healthy and not at risk of developing disease such as a neurological disorder, and they can receive no therapeutic benefit; however they receive benefit concerning performance improvement in general and a perception of well-being from the method, especially in connection with brain functions. Preferably, the performance improvement and / or perception of well-being is more effective than the human would receive from 2’-FL consumption. In other embodiment, the human may be in the state or at risk of developing a neurological disorder. In this case the method can lead to, in connection to the neurological disorder, the preservation or protection of neurons and neural tissue, enhancement of the production of neurotransmitters, supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system. In any aspect of the invention disclosed above, the recommended dose is in the range from about 200 mg to about 20 g per day, in certain embodiments from about 300 mg to about 15 g per day, from about 400 mg to about 10 g per day, in certain embodiments from about 500 mg to about 10 g per day, in certain embodiments from about 1 g to about 10 g per day.

[0090] Whilst the invention has been described with reference to embodiments, it will be appreciated that various modifications are possible within the scope of the invention.

[0091] The following numbered aspects of the invention are provided:

[0092] 1. LNDFH-I or a synthetic composition comprising LNDFH-I for use in: the preservation or protection of neurons and neural tissue, enhancing the production of neurotransmitters, supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human patient being in the state or at risk of developing a neurological disorder.

[0093] 2. The synthetic composition for the use of aspect 1 , which is a nutritional composition.

[0094] 3. The synthetic composition for the use of aspect 1 , which is a pharmaceutical composition.

[0095] 4. The synthetic composition for the use of any of the aspects 1 to 3, which is a not a mammalian milk.

[0096] 5. The synthetic composition for the use of any of the aspects 1 to 3, which is a not a processed mammalian milk.

[0097] 6. A method for the preservation or protection of neurons and neural tissue of a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, optionally in a form of a synthetic composition comprising LNDFH-I.

[0098] 7. A method for enhancing the production of neurotransmitters in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, optionally in a form of a synthetic composition comprising LNDFH-I.

[0099] 8. A method for supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, optionally in a form of a synthetic composition comprising LNDFH-I. 9. A method for supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system in a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, optionally in a form of a synthetic composition comprising LNDFH-I.

[0100] 10. A method for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a human, the method comprising orally or enterally administering to the human an effective amount of LNDFH-I, optionally in a form of a synthetic composition comprising LNDFH-I.

[0101] 11. The method of any of the aspects 6 to 9, wherein the amount of LNDFH-I is effective to enhance the production of one or more metabolites selected from the group consisting of N- acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid.

[0102] 12. The method of aspect 10 or 11, wherein the production of N-acetyl-L-tyrosine is enhanced.

[0103] 13. The method of aspect 10 or 11, wherein the production of N-acetyl-L-glutamine is enhanced.

[0104] 14. The method of aspect 10 or 11, wherein the production of nicotinic acid is enhanced.

[0105] 15. A non-therapeutic use of LNDFH-I for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a healthy human.

[0106] 16. The non-therapeutic use of aspect 15, wherein the production of N-acetyl-L-tyrosine is enhanced.

[0107] 17. The non-therapeutic use of aspect 15, wherein the production of N-acetyl-L-glutamine is enhanced.

[0108] 18. The non-therapeutic use of aspect 15, wherein the production of nicotinic acid is enhanced.

[0109] 19. A non-therapeutic use of LNDFH-I for protection of neurons and neural tissue in a healthy human.

[0110] 20. The non-therapeutical use of LNDFH-I of aspect 19, wherein the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of the human is enhanced.

[0111] 21. The non-therapeutic use of aspect 19 or 20 to enhance brain performance and perception of well-being. 22. The non-therapeutic use of any of the aspects 15 to 21 , wherein LNDFH-I is comprised in a synthetic composition.

[0112] 23. The non-therapeutic use of aspect 22, wherein the synthetic composition is a nutritional composition.

[0113] 24. The non-therapeutic use of aspect 23, wherein the nutritional composition is a not a mammalian milk.

[0114] 25. The non-therapeutic use of aspect 23, wherein the nutritional composition is a not a processed mammalian milk.

[0115] 26. The method of any of the aspects 6 to 14, wherein the amount of LNDFH-I is in the range of from about 200 mg to about 20 g per day.

[0116] 27. The method of aspect 26, wherein the amount of LNDFH-I is in the range of from about 300 mg to about 15 g per day.

[0117] 28. The method of aspect 27, wherein the amount of LNDFH-I is in the range of from about 400 mg to about 10 g per day.

[0118] 29. The method of aspect 28, wherein the amount of LNDFH-I is in the range of from about 500 mg to about 10 g per day.

[0119] 30. The method of aspect 29, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

[0120] 31. The non-therapeutic use of any of the aspects 15 to 25, wherein the amount of LNDFH-I is in the range of from about 200 mg to about 20 g per day.

[0121] 32. The method of aspect 31 , wherein the amount of LNDFH-I is in the range of from about 300 mg to about 15 g per day.

[0122] 33. The method of aspect 32, wherein the amount of LNDFH-I is in the range of from about 400 mg to about 10 g per day.

[0123] 34. The method of aspect 33, wherein the amount of LNDFH-I is in the range of from about 500 mg to about 10 g per day.

[0124] 35. The method of aspect 34, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

[0125] 36. LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to the any of the aspects 1 to 5, wherein the amount of LNDFH-I is in the range of from about 200 mg to about 20 g per day. 37. LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to aspect 36, wherein the amount of LNDFH-I is in the range of from about 300 mg to about 15 g per day.

[0126] 38. LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to aspect 37, wherein the amount of LNDFH-I is in the range of from about 400 mg to about 10 g per day.

[0127] 39. LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to aspect 38, wherein the amount of LNDFH-I is in the range of from about 500 mg to about 10 g per day.

[0128] 40. LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to aspect 39, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

[0129] 41. The non-therapeutic use of LNDFH-I according to the any of the aspects 15 to 25, wherein the amount of LNDFH-I is in the range of from about 200 mg to about 20 g per day.

[0130] 42. The non-therapeutic use of LNDFH-I according to aspect 41, wherein the amount of LNDFH-I is in the range of from about 300 mg to about 15 g per day.

[0131] 43. The non-therapeutic use of LNDFH-I according to aspect 42, wherein the amount of LNDFH-I is in the range of from about 400 mg to about 10 g per day.

[0132] 44. The non-therapeutic use of LNDFH-I according to aspect 43, wherein the amount of LNDFH-I is in the range of from about 500 mg to about 10 g per day.

[0133] 45. The non-therapeutic use of LNDFH-I according to aspect 44, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

[0134] EXAMPLES

[0135] Example 1

[0136] A kinetic, ex vivo study was implemented, simulating the colonic fermentation of test products by the gut microbiota derived from healthy infants (n = 12). Individual bioreactors were processed in parallel in a bioreactor management device (Cryptobiotix, Ghent, Belgium). Each bioreactor contained 5 ml of nutritional medium with faecal inoculum, dosed with the test product and sealed individually before being rendered anaerobic. After preparation, bioreactors were incubated under continuous agitation (140 rpm) at 37 °C for 24 h. Upon gas pressure measurement in the headspace, liquid samples were collected for subsequent analysis. Test products were HMOs (2’-FL, LNFP-I, LNDFH-I), at the concentration 5 g / l, simulating intake of 5 g per day. 12 samples per study arm were pooled to a single sample and were then analysed in technical triplicate (n = 3). The metabolites were identified with LC-MS by retention times and accurate mass.

[0137] The LC-MS analysis was carried out using a Thermo Scientific Vanquish LC coupled to Thermo Q Exactive HF MS. An electrospray ionization interface was used as ionization source. Analysis was performed in negative and positive ionization mode. The LIPLC was performed using a slightly modified version of the protocol described by Doneanu et al.

[0138] (https: / / www.waters.com / webassets / cms / library / docs / 720004042en.pdf.). Peak areas were extracted using Compound Discoverer 3.1 (Thermo Scientific). In addition to the automatic compound extraction by Compound Discoverer 3.1, a manual extraction of compounds included in an in-house library was performed using Skyline 21.1 (MacCoss Lab Software).

[0139] Figure 1. depicts the Area Under Curve (AUC) values determined in each arm. AUC is a measure of the concentration of the compound it represents. This area value was integrated based on the peak of the metabolite in the chromatograms of the LC-MS analysis. The bold circle is the mean value of the triplicates in each measurement. From Figure 1 it can be concluded that LNFP-I enhanced the production of metabolites not only compared to blank (NSC) but compared to 2’-FL, the most abundant HMO, and to the structurally related HMO LNFPI-I, too. Therefore, LNDFH-I is a more promising candidate to promote brain health.

Claims

CLAIMS1 . A non-therapeutic use of lacto-N-difucohexaose-l (LNDFH-I) to enhance brain performance and perception of well-being in a healthy human.

2. The non-therapeutical use according to claim 1 , wherein the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L- glutamine, and nicotinic acid in the gastrointestinal tract of the human is enhanced.

3. The non-therapeutical use according to claim 1 or 2, wherein the human is elderly individual above 60 years of age, and enhancing brain performance and perception of well-being comprises improving cognitive functions.

4. The non-therapeutical use according to claim 1 or 2, wherein the human is an athlete, student or headworker, and enhancing brain performance and perception of well-being comprises optimizing cognitive functions.

5. The non-therapeutical use according to claim 1 or 2, wherein the human is an infant and perception of well-being comprises optimizing the growth, differentiation and maturation of the nervous system.

6. A non-therapeutic use of lacto-N-difucohexaose-l (LNDFH-I) for enhancing the production of one or more metabolites selected from the group consisting of N-acetyl-L- tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of a healthy human.

7. The non-therapeutic use of any of the precedent claims, wherein LNDFH-I is comprised in a synthetic composition.

8. The non-therapeutic use of claim 7, wherein the synthetic composition is a nutritional composition.

9. The non-therapeutic use of claim 8, wherein the nutritional composition is a not a mammalian milk.

10. The non-therapeutic use of claim 9, wherein the nutritional composition is a not a processed mammalian milk.11 . The non-therapeutic use of any of the precedent claims, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

12. Lacto-N-difucohexaose-l (LNDFH-I) or a synthetic composition comprising LNDFH-I for use in: the preservation or protection of neurons and neural tissue, enhancing the production of neurotransmitters,supporting or improving one or more of cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, and / or supporting or improving the biological processes that govern the growth, differentiation, and maturation of the nervous system, in a human patient being in the state or at risk of developing a neurological disorder.

13. The LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to claim 12, wherein the production of one or more metabolites selected from the group consisting of N-acetyl-L-tyrosine, N-acetyl-L-glutamine, and nicotinic acid in the gastrointestinal tract of the human is enhanced.

14. The synthetic composition for the use according to claim 12 or 13, which is a not a mammalian milk or a processed mammalian milk.

15. The LNDFH-I for the use or the synthetic composition comprising LNDFH-I for the use according to any of the precedent claims, wherein the amount of LNDFH-I is in the range of from about 1 g to about 10 g per day.

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