RU2791622C2 - Composition containing brest milk oligosaccharides (bmo) for use in improving short-term memory and obtaining other cognitive benefits - Google Patents

Abstract

FIELD: nutritional composition containing breast milk oligosaccharide (BMO).

SUBSTANCE: use of a nutritional composition containing breast milk oligosaccharide (BMO) for improving or activating declarative memory in an infant or young child. Wherein the composition contains breast milk oligosaccharides 2’-fucosyllactose (2’-FL) and lacto-N-neotetraose (LNnT). The composition additionally contains cow milk oligosaccharides 3’-sialyllactose and 6’- sialyllactose and galactooligo-saccharide (GOS).

EFFECT: improvement or activation of declarative memory in an infant or young child

7 cl, 1 dwg, 3 tab, 2 ex

Description

Prerequisites for the creation of the invention

The present invention relates to a composition for use in improving short term memory and/or developing optimal short term memory and other associated cognitive benefits, particularly in infants and young children.

The present invention also relates per se to the field of nervous system health, nervous system protection and nervous system development. The invention particularly relates to the administration of human breast milk oligosaccharides (HMOs) to induce the development of optimal short term memory and/or improve short term memory and/or promote healthy cognitive development in infants and young children. The invention can be particularly applied to weak or premature babies.

The central nervous system (CNS), and in particular the brain, controls cognitive functions. The cerebral cortex, which is a layer of nervous tissue at the periphery of the mammalian brain, plays a key role in attention, perceptual perception, higher cognitive function (executive function), and the integration of information from afferent impulses.

The development and maturation of the central nervous system is an extremely complex biological phenomenon that involves many physiological processes, including, for example, the growth and differentiation of neurons and glial cells, neuronal guidance and branching, and the formation of interneuronal interaction (nerve signals) through axon growth and synaptogenesis.

Neuronal plasticity, defined as the ability of the brain to continually adapt function and structural organization to changing demands, is important for neural maturation and mature function. It is essential for the proper functioning of the brain and is necessary for the processes of cognition, learning and memory. Several of the neuronal markers, including proteins and neurotrophic factors such as brain-derived neurotrophic factor (BDNF), required for or at least involved in these physiological processes have been identified and investigated in the literature [Huang, E.J. and Reichardt, L. F. (2001); Neurotrophins: Roles in Neuronal Development and Function, Annu. Rev. Neurosci., 24: 677-736]; [Musumeci, G. and Minichiello, L. (2011); BDNF-TrkB signaling in fear learning: from genetics to neural networks, Rev. Neurosci., 22(3):303–15]; [Xiao, J. et al. (2009); The role of neurotrophins in the regulation of myelin development, Neurosignals, 17: 265–276] and [Von Bohlen and Halbach, O. (2011); Immunohistological markers for proliferative events, gliogenesis, and neurogenesis within the adult hippocampus, Cell Tissue Res., 345(1):1–19].

The central nervous system develops immediately after conception, during pregnancy, and continues to develop until early adulthood. In particular, structural maturation predominantly occurs before birth, while functional network maturation predominantly occurs after birth. In human embryos, the cerebral cortex develops quite late and over a long period of time.

In the intrauterine period, a person experiences a peak of maturation and growth of neurons / brain from the 30th week of pregnancy.

Developing the ability to temporarily store and manage information (such as visual cues), called short-term memory, is an important step in the development of cognitive function in infants and young children. The development and/or improvement of short-term memory is closely related to neuronal plasticity. While this development and improvement of short-term memory is of particular importance during the first months/years of life (when neuronal plasticity is highest), it also affects older subjects, adolescents and adults. Short-term memory deteriorates with aging, both in healthy older people and older people suffering from illnesses.

Premature babies, by definition, are born with still underdeveloped brains, and they really show only basic electrical activity in the primary sensory areas of the cerebral cortex, that is, in those areas that are responsible for touch, sight and hearing, as well as in primary motor areas. cerebral cortex. In these children, postnatal gradual brain maturation is necessary to compensate for the state of cerebral immaturity at birth, and such compensatory maturation is especially important for the more complex part of the brain that mediates much of the emotional, social, and cognitive maturation in the first few years of life [Lubsen, J et al. (2011); Microstructural and functional connectivity in the developing preterm brain, Seminars in Perinatology, 35, 34–43].

Premature babies are born at a time that is very important for the structural and functional development and maturation of the brain, and thus they skip the stage of intrauterine brain development. After birth, they are at risk for pathological conditions, including hemorrhagic and hypoxic-ischemic brain lesions, as well as the risk of developmental problems later in life, including cognitive impairment. This risk appears to be higher the more premature babies are born and the lower their birth weight. Cognitive impairment associated with lower IQ, reduced attention span and short-term memory, and problems with executive functions may persist into school and adolescence [Talge, N. et al. (2010). Late-Preterm Birth and its Association with Cognitive and Socioemotional Outcomes at 6 Years of Age. Pediatrics, 126, 1124–1131; van Baar, A., et al. (2009). Functioning at school age of moderately preterm children born at 32 to 36 weeks' gestational age. Pediatrics, 124, 251–257; Farooqi, A et al. (2011). Impact at age 11 years of major neonatal morbidities in children born extremely preterm. Pediatrics, 127, e1247–1257; Nosarti, C. et al. (2010). Neurodevelopmental outcomes of preterm birth. Cambridge: Cambridge University Press].

In general, CNS immaturity or delayed CNS maturation may occur in infants such as:

premature infants, low birth weight infants (< 2500 g), very low and extremely low birth weight infants (< 1500 g), extremely low birth weight infants (< 1000 g), and infants small for their gestational age ( SGA); [Allen, M.C. (2008); Neurodevelopmental outcomes of preterm infants, Curr. Opin Neurol., 21(2): 123–8];

preterm or full-term infants with intrauterine growth retardation (IUGR), which occurred as a result of any adverse events during pregnancy (maternal smoking, maternal medication, poor quality of the placenta, malposition of the placenta, malnutrition of the mother and fetus, excessive stress / mother's anxiety, etc.); [Gregory, A. et al. (2008); Intrauterine Growth Restriction Affects the Preterm Infant’s Hippocampus, Pediatric Research, 63(4): 438–443];

any newborn and infant with neurodevelopmental retardation as a result of, for example, hypoxemia (ischemia) at delivery, postnatal complications, postnatal steroid treatment, or any other adverse event [Barrett, R.D. et al. (2007); Destruction and reconstruction: hypoxia and the developing brain, Birth Defects Res. C. Embryo Today, 81: 163–76].

These infants show cognitive dysfunction as well as growth and developmental dysfunction, indicating that the neurodevelopmental process has not been "compensated" enough. Immaturity or delayed maturation of the cerebral cortex can lead to slow and/or impaired ability to learn, generalize information, process afferent impulses, loss or insufficient development of higher functions of thinking, executive functions, concentration, attention, motor skills and language. This can lead to behavioral problems, a pathological decrease in intelligence and, consequently, a pathological decrease in mental performance.

It has generally been noted that breastfeeding premature infants can lead to improved neuronal development compared to formula feeding. (See, for example: Rozé et al. The apparent breastfeeding paradox in very preterm infants: relationship between breast feeding, early weight gain and neurodevelopment based on results from two cohorts, EPIPAGE and LIFT. BMJ Open 2012;2:e000834. doi: 10.1136/bmjopen-2012-000834).

According to the inventors, this generally indicates a possible lack of some of the nutrients present in human breast milk in formula or delivery in a suboptimal amount. There is a need to identify key differences between traditional formula and human breast milk and to tailor artificial formula accordingly.

Behavioral and neurodevelopmental disorders associated with delayed maturation of the cerebral cortex include attention deficit/hyperactivity disorder and autism spectrum disorders.

A person's cognitive function can be measured using clinical tests that are age dependent; pediatricians and child development specialists are familiar with many such tests. For infants and infants, there are screening and neurodevelopmental tests such as the Bailey Scale for Infant Development (BSID), the Brazelton Behavioral Assessment Scale for Newborns, the Neuropsychological Developmental Assessment (NEPSY), and the Griffiths Scale for Mental Development. For preschool and/or school-age children, cognitive ability tests include the Peabody Picture Vocabulary Test (PPVT), the Non-Verbal Intelligence-2 Test (TONI-2), the Wechsler Preschool and Early Childhood Intelligence Scale (WPPSI), and the Colored Progressive Raven matrices (CPM).

One aspect of cognitive development can be tested by testing short-term memory. Such tests are known in the art. They can provide a direct indication of the ability to recognize signals (short-term) and are also indicators of the general cognitive development of the subject. These tests, known in the art, are capable of distinguishing short term memory from another form of memory, such as long term memory. Memory tests are commonly available and are described in particular by Ross-Sheehy, S. et al. (2003); The development of visual short-term memory capacity in infants. Child Dev, 74(6):1807–22 and Gathercole (1999), Cognitive approaches to the development of short-term memory TICS, 3(11):410–419.

Nutrition is known to play an important role in the maturation of neurons in the brain (reviewed [Huppi, P. S. (2008); Nutrition for the Brain, Pediatric Research, 63(3): 229–231 and Cusik and Georgieff (2016); The Role of Nutrition in Brain Development: The Golden Opportunity of the “First 1000 Days”, Journal of Pediatrics, 175:16–21).

The consequences of malnutrition can be irreversible and may include reduced cognitive development, impaired memory and learning ability, and hence future economic productivity. [Horton, R; (2008) The Lancet, Vol. 371, Issue 9608, page 179; [Laus, M.F. et al. (2011); Early postnatal protein-calorie malnutrition and cognition: a review of human and animal studies, Int. J. Environ. Res. Public Health., 8(2): 590–612].

Although breast milk is known to provide the best nutritional support for the developing brain, if breastfeeding is not possible, there is a need to provide an artificial nutritional composition (such as infant formula or formula for feeding babies) that induces improvement or activates the development of optimal cognitive functions.

Thus, oral administration is a suitable way to positively influence the development of the nervous system to ensure optimal development of cognitive function, memory and mental abilities in preterm or term newborns, infants, toddlers, children of older or adolescent age or young animals.

However, there is little known and reliable data on the ability of nutritional diets or nutritional compositions to influence the development or activation of memory, in particular short-term memory, and in particular in infants and young children.

Abbott Laboratories inc. publication EP0975235B1. describes an artificial nutritional composition containing one or more human milk oligosaccharides, wherein the HMOs in the composition are selected from the group consisting of eight HMOs (3-fucosyllactose, lacto-N-fucopentaose III, lacto-N-fucopentaose II, difucosyllactose (diFL), 2 '-fucosyllactose, lacto-N-fucopentaose I, lacto-N-neotetraose and lacto-N-fucopentaose V), wherein said composition is intended for use in cases of normal, healthy infants, children, adults, or individuals with special needs, such as those associated with certain pathological conditions. This European Patent states that, in general, oligosaccharides protect infants from viral and bacterial infections of the respiratory, gastrointestinal and genitourinary tracts.

WO2012092160 and US2015231159A1 by the same applicant (Abbott laboratories inc.) provide a link between HMO and the growth of beneficial bacteria and hypothesize a link to cognition.

There is still a need to activate and support the healthy development of cognitive function in general, and in particular short-term and long-term memory.

There is a need to improve such memory functions in young subjects, in particular infants and young children.

There is a need to activate the development of such memory functions in infants and young children.

There is a need to provide nutritional compositions for infants and young children that improve short term memory and/or enhance and activate short term memory development.

There is a need to activate and/or improve the performance of short-term memory independent of long-term memory.

There is a need to provide such activation, improvement or enhancement of short term memory and/or working memory and/or declarative memory through nutritional interventions, especially in infants and young children.

There is a need to provide such nutritional intervention and/or preventive nutritional intervention in a form that is acceptable to a population of subjects, in particular those individuals in that population who are most vulnerable or most in need. In addition, there is a need to avoid inducing adverse events, side effects or adverse effects in such a population.

There is a need to provide a population of subjects in need with such solutions in the simplest and most economical way, preferably not through the use of specific ingredients considered drugs or medicaments, but preferably as part of the diet.

The present invention relates to all mammals, including animals and humans, and in particular to infants and young children, in which brain plasticity is the highest.

Statement of the Invention

The present inventors have unexpectedly found that the administration of a particular oligosaccharide, in particular breast milk oligosaccharides, is particularly effective in improving and/or activating and/or improving short-term memory and/or declarative memory and/or working memory, in particular in infants. and/or young children. The introduction can be carried out as part of a nutritional intervention.

Brief description of graphic materials

On FIG. 1 shows the effect of different diets in piglets on a recognition test as an indicator of working memory; recognition coefficient when using intertest intervals of 1 hour or 2 days with different diets [CON (control); OGM (2 FL + LNnT); BMO (cow's milk oligosaccharide); OGM + BMO]. The use of HMO alone resulted in a statistically significant increase in recognition memory with a 1-hour intertest interval, but unexpectedly did not improve recognition memory over a 2-hour intertest interval. This indicates a clear positive effect of HMO (2' FL + LNnT) on short-term recognition memory. The use of HMO + BMO leads to a numerical increase in recognition memory with a 1-hour intertest interval and a statistically significant increase in recognition memory with a 24-hour intertest interval.

Detailed description of exemplary embodiments

DEFINITIONS

In this document, the following terms have the following definitions.

The term "infant" means a child under the age of 12 months.

The term "young child" means a child between one and three years of age.

As used herein, the term "oligofructose" (abbreviated OF) refers to a fructose oligomer (i.e., fructose oligosaccharide) having a degree of polymerization of 2 to 10, such as a degree of polymerization of 2 to 8. Oligofructose may also be referred to as fructooligosaccharides (abbreviated FOS) or short chain fructooligosaccharides (scFOS for short). In this document, the terms "oligofructose" (OF), "fructose oligosaccharide" (FOS), "fructooligosaccharide" (FOS), "short chain fructooligosaccharide" (scFOS) have the same meaning and can be used interchangeably.

Inulin, which is a long chain polymer, is specifically excluded from this OF definition. Oligofructose is distinguished from inulin by the degree of polymerization (inulin has much longer chains).

FOS/scFOS/oligofructose are generally commercially available, eg under the tradename ORAFTI oligofructose from Beneo GmbH (Mannheim, Germany) (eg ingredient Orafti® P95).

The term "human milk oligosaccharide" is used as an abbreviation for HMO and collectively refers to oligosaccharides present in human milk and falls under the standard definition accepted by any person skilled in the art. OGMs are, without limitation, fucosylated oligosaccharides (eg, diFL, 2FL), lacto-N tetraose (LNT), lacto-N-neotetraose (LNnT), or sialylated oligosaccharides.

The term "cow's milk oligosaccharide" (abbreviation BMO) refers to oligosaccharides present in cow's milk and falls under the standard definition accepted by any person skilled in the art. The mixture of cow's milk oligosaccharides (BMOS), which is used in the context of the present invention, was obtained from cow's milk whey. Briefly, the bovine serum ultrafiltrate, including oligosaccharides such as 3'- and 6'-sialyllactose and GOS, was demineralized by a combination of electrodialysis and ion exchange as described in the art.

As used herein, the term "sn-2-palmitate" refers to palmitic acid at the sn-2 position of the triglyceride to which it is associated.

The term "short-term memory" (STM) refers to a system for temporarily storing and managing information needed to perform complex cognitive tasks such as learning, thinking, and understanding. Short-term memory is needed to recognize cues such as visual cues.

Short-term memory (STM) allows you to remember something for a period of a few seconds to an hour without repetition. Short-term memory encodes, for example, auditory information, is supported by temporal neural communication structures, and depends on areas of the frontal lobe (especially the dorsolateral prefrontal cortex) and parietal lobe in which elements are stored temporarily.

The term "infant formula" means a food product specifically intended for use by infants during the first four to six months of life and which by itself satisfies the nutritional needs of this category of persons (Article 1.2 of European Commission Directive 91/321/EEC of 14 May 1991 about infant formulas and mixtures for feeding children).

The term "formula for infants" means a food product specifically intended for consumption by infants over four months of age and which constitutes the main liquid component in the diet of this category of individuals, which involves progressively increasing diversity.

The term "initial infant formula" means a food product specifically intended for consumption by infants during the first four months of life.

Infant formula, formula for feeding infants and initial infant formula can be either in the form of a liquid, ready-to-drink or concentrated, or in the form of a dry powder, which can be reconstituted into formula after the addition of water. Such mixtures are well known in the art.

The term "infant food" means a food product specifically intended for consumption by infants during the first years of life.

The term "cereal-based infant composition" means a food product specifically intended for use by infants during the first years of life.

The term "milk formula for children from 1 to 3 years" means a milk-based drink adapted to the specific nutritional needs of young children.

The term "weaning period" means the period during which mother's milk is replaced in the infant's diet with another food.

The term "nutritional composition" means the composition that feeds the subject. This nutritional composition is usually intended for oral or intravenous use and usually includes a lipid or fat source and a protein source. The nutritional composition is preferably a complete nutritional formula that meets all or most of the nutritional needs of the subject (eg, infant formula).

The term "artificial mixture" means a mixture obtained by chemical and/or biological means, which may be chemically identical to a mixture of natural origin, present in the milk of mammals.

The term "sialylated oligosaccharide" means an oligosaccharide having a sialic acid residue.

The term "fucosylated oligosaccharide" means an oligosaccharide having a fucose residue.

The term "prebiotic" means non-digestible carbohydrates that favorably affect the host by selectively stimulating the growth and/or activity of beneficial bacteria, such as bifidobacteria, in the human colon (Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics J Nutr 1995;125:1401–12).

The term "probiotic" means preparations of microbial cells or components of microbial cells that have a beneficial effect on the health or well-being of the host organism. (Salminen S, Ouwehand A. Benno Y. et al. Probiotics: how should they be defined Trends Food Sci. Technol. 1999:10 107–10).

Unless otherwise indicated, all percentages are wt%.

If the amounts of ingredients are given as mass of ingredient/weight of powdered nutritional composition, it is also assumed that the present invention also refers to the corresponding quantity per liter, considering the dilution factor of the dry powdered nutritional composition of 130 g/l (or otherwise specified in the dilution instructions).

breast milk oligosaccharides

Human milk oligosaccharides (HMOs) are collectively the third largest solids component in breast milk after lactose and fat. HMO usually consists of lactose at the reducing end with a carbohydrate core that often contains fucose or sialic acid at the non-reducing end. Approximately one hundred milk oligosaccharides have been isolated and characterized, but they represent only a very small part of the total number to be characterized.

Infant formulas have previously been developed for various purposes using HMO ingredients such as fucosylated oligosaccharides, lacto-N-tetraose, lacto-N-neotetraose, or sialylated oligosaccharides.

The composition of the invention may contain 2'-fucosyllactose (2FL, also abbreviated as 2'FL) and/or N-acetyllactosamine, such as lacto-N-neotetraose (LNnT) or lacto-N-tetraose (LNT).

In one embodiment, the nutritional composition according to the invention comprises a human milk oligosaccharide selected from the list comprising N-acetyllactosamine, sialylated oligosaccharides, fucosylated oligosaccharides, 2FL, LNnT, diFL, LNT, or a combination thereof.

N-acetyllactosamine

In some embodiments, the implementation of the composition of the invention contains at least one N-acetyllactosamine. In other words, the composition according to the invention contains N-acetyllactosamine and/or an oligosaccharide containing N-acetyllactosamine. Suitable oligosaccharides containing N-acetyllactosamine include lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

Thus, according to the invention, N-acetyllactosamine is preferably selected from the group consisting of lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

LNT and LNnT can be chemically synthesized by enzymatic transfer of saccharide units from donor moieties to acceptor moieties using glycosyltransferases, as described, for example, in US Pat. Nos. 5,288,637 and WO 96/10086. In an alternative embodiment, LNT and LNnT can be produced by chemically converting ketohexoses (e.g., fructose), either free or bound to an oligosaccharide (e.g., lactulose), to an N-acetylhexosamine or N-acetylhexosamine-containing oligosaccharide, as described in Wrodnigg, T.M. ; Stutz, A.E. (1999) Angew. Chem. Int. Ed. 38:827-828. The N-acetyllactosamine thus obtained can then be transferred to lactose as an acceptor functional group.

The composition according to the invention preferably contains from 0.1 to 3 g of N-acetyllactosamine per 100 g of the composition, based on the weight of dry matter. It preferably contains from 0.1 to 3 g LNnT per 100 g of the composition, based on the weight of dry matter.

In one embodiment, the nutritional composition according to the invention comprises N-acetyllactosamine, preferably selected from the group consisting of lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

Siallylated oligosaccharides

The composition in accordance with the invention in some embodiments, the implementation may contain one or more sialylated oligosaccharides.

The sialylated oligosaccharides may be selected from the group consisting of 3'-sialyllactose and 6'-sialyllactose. Preferably, both 3'-sialyllactose and 6'-sialyllactose are present in said composition. In this embodiment, the ratio of 3'-sialyllactose to 6'-sialyllactose may be between 5:1 and 1:2.

The 3' and 6' forms of sialyllactose can be isolated from a natural source such as animal milk by chromatography or filtration. In an alternative embodiment, they can be produced by a biotechnological agent using specific sialyl transferases or sialidases, neuraminidase by enzyme-based fermentation (recombinant or natural enzymes), by chemical synthesis or by microbiological fermentation. In the latter case, microorganisms can express their natural enzymes and substrates, or it is possible to create microorganisms that produce the corresponding substrates and enzymes. You can use cultures of microorganisms of the same species or mixed cultures. The formation of sialyl oligosaccharides can be initiated by acceptor substrates starting at any degree of polymerization (DP) from DP = 1 onwards. In an alternative embodiment, sialyllactoses can be obtained by chemical synthesis from lactose and free N'-acetylneuraminic acid (sialic acid). Sialyl lactoses are also commercially available, for example from Kyowa Hakko Kogyo, Japan.

The composition according to the invention preferably contains from 0.05 to 2 g of sialylated oligosaccharide(s), for example from 0.1 to 2 g of sialylated oligosaccharide(s) per 100 g of composition, based on mass of dry matter.

In one embodiment, the nutritional composition according to the invention comprises a sialylated oligosaccharide, preferably selected from the group consisting of 3'-sialyllactose and 6'-sialyllactose. More preferably, said composition contains both 3'-sialyllactose and 6'-sialyllactose, with the ratio of 3'-sialyllactose and 6'-sialyllactose being preferably in the range of 5:1 to 1:2.

Fucosylated oligosaccharide

The composition according to the present invention may contain one or more fucosylated oligosaccharides. The fucosylated oligosaccharides preferably consist of or contain 2'-fucosyllactose (2-FL).

The fucosylated oligosaccharide may be selected from the group consisting of 2'-fucosyllactose, 3-fucosyllactose, difucosyllactose, lacto-N-fucopentaose (i.e., lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III and lacto-N-fucopentaose V), lacto-N-difucohexaose I, fucosyl lacto-N-hexaose, difucosyl lacto-N-hexaose I, and difucosyl lacto-N-neohexaose II. A particularly preferred fucosylated oligosaccharide is 2'-fucosyllactose (2FL).

The fucosylated oligosaccharide can be isolated from a natural source such as animal milk by chromatography or filtration. In an alternative embodiment, it can be produced by biotechnological means using specific fucosyltransferases and/or fucosidase, either through the use of an enzyme-based fermentation method (recombinant or natural enzymes) or a microbiological fermentation method. In the latter case, microorganisms can express their natural enzymes and substrates, or it is possible to create microorganisms that produce the corresponding substrates and enzymes. Single microorganism cultures and/or mixed cultures can be used. Fucosylated oligosaccharide formation can be initiated by acceptor substrates starting at any degree of polymerization (DP) from DP=1 onwards. In an alternative embodiment, fucosylated oligosaccharides can be obtained by chemical synthesis from lactose and free fucose. Fucosylated oligosaccharides are also commercially available, for example from Kyowa Hakko Kogyo, Japan.

The composition according to the invention preferably contains from 0.1 to 3 g of fucosylated oligosaccharide(s) per 100 g of composition on a dry weight basis, most preferably 2FL

In one embodiment, the nutritional composition of the invention comprises a fucosylated oligosaccharide, preferably selected from the group consisting of 2'-fucosyllactose, 3-fucosyllactose, difucosyllactose, lacto-N-fucopentaose (i.e., lacto-N-fucopentaose I, lacto-N- fucopentaose II, lacto-N-fucopentaose III and lacto-N-fucopentaose V), lacto-N-difucohexaose I, fucosyl lacto-N-hexaose, difucosyl lacto-N-hexaose I and difucosyl lacto-N-neohexaose II, and preferably the fucosylated oligosaccharide is is 2'-fucosyllactose (2-FL).

Additional Prebiotics

In addition to the main oligosaccharides that are the subject of this patent, the composition of the invention may additionally contain at least one or one additional prebiotic, usually in an amount of from 0.3 to 10% by weight of the composition.

Prebiotics are generally indigestible in the sense that they are not broken down or absorbed in the stomach or small intestine and thus remain native when they pass to the large intestine where they are selectively fermented by beneficial bacteria.

The composition in accordance with the invention may contain, in some embodiments, the implementation of oligofructose (OF). An example of such an OF is the commercial ingredient ORAFTI® manufactured by Beneo GmbH (Mannheim, Germany).

In some embodiments, the prebiotic compositions of the invention comprise other fructooligosaccharides (FOS) and/or galactooligosaccharides (GOS). A combination of prebiotics such as 90% HOS and 10% short chain fructooligosaccharides can be used, such as the product sold by BENEO-Orafti under the brand name Orafti® oligofructose (see http://www.beneo-orafti.com/Our-Products). /Oligofructose) (formerly Raftilose®), or 10% inulin, such as the product sold by BENEO-Orafti under the brand name Orafti® inulin (see http://www.beneo-orafti.com/Our-Products/Inulin ) (formerly Raftiline®). Another combination of prebiotics is 70% short chain fructooligosaccharides and 30% inulin, which is a product sold by BENEO-Orafti® under the brand name Prebio 1.

In one embodiment, the nutritional composition according to the invention contains a prebiotic selected from the list of bovine milk oligosaccharides, inulin, xylooligosaccharides, polydextrose, or any combination thereof.

In one embodiment, the nutritional composition according to the invention comprises a cow's milk oligosaccharide, wherein said cow's milk oligosaccharides are N-acetylated oligosaccharide, galactooligosaccharide, sialylated oligosaccharide, fucosylated oligosaccharide, or a combination thereof.

Probiotics

The composition of the invention may further comprise at least one probiotic bacterial strain, said probiotic bacterial strain being preferably Bifidobacteria and/or Lactobacilli.

Suitable probiotic bacterial strains include Lactobacillus rhamnosus ATCC 53103 available from Valio Oy, Finland under the trademark LGG; Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus paracasei CNCM I-2116, Lactobacillus johnsonii CNCM I-1225, Streptococcus salivarius DSM 13084 sold by BLIS Technologies Limited, New Zealand under the name KI2; Bifidobacterium lactis CNCM 1-3446, sold inter alia by Christian Hansen, Denmark, under the trademark Bb 12; Bifidobacterium longum ATCC BAA-999 sold by Morinaga Milk Industry Co. Ltd., Japan, under the trademark BB536; Bifidobacterium breve sold by Danisco under the trademark Bb-03; Bifidobacterium breve sold by Morinaga under the trademark M-16V; Bifidobacterium infantis marketed by Procter & Gambie Co. under the Bifantis trademark; and Bifidobacterium breve sold by Institut Rosell (Lallemand) under the trademark R0070.

The composition according to the invention preferably contains from 10e3 to 10e12 cfu of the probiotic bacterial strain, more preferably from 10e7 to 10e12 cfu per gram of the composition, based on the weight of dry matter.

In one embodiment, the nutritional composition comprises a probiotic bacterial strain selected from a list consisting of Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus lactis, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus bulgari, Lactococcus lactis, Lactococcus diacetylactis, Lactococcus cremoris, Streptococcus salivarius, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis or Bifidobacterium adolescentis, or any mixture thereof.

Target population

The present invention is directed to infants and young children.

In one embodiment, the present invention is directed to young mammals such as infants (eg, 0 to 6 months or 0 to 12 months), toddlers (eg, 1 to 3 years or 1 to 7 years). Without being bound by any theory, young mammals have high brain plasticity and brain development (or brain connectivity) and benefit most from the present invention. In addition, it is contemplated that the present invention may target specific nutrition intervention windows (e.g., children aged 6 months to 3 years, 3 months to 18 months), subpopulations with special needs (vulnerable young mammals; senile and semi-senile mammals) or in the recovery phase.

According to a preferred embodiment, the composition according to the invention is for use in healthy infants and/or healthy young children. In one embodiment, the invention is of particular value to frail infants, premature infants, and/or infants born with abnormally low birth weight and/or infants suffering from intrauterine growth retardation. The preferred period of application is (s) the period (s) of the most rapid development of memory and / or development of brain connections.

The composition of the invention is intended for infants and/or young children 7 years of age or less, preferably 3 years of age or less, most preferably 1 year of age or less. In one embodiment, the composition is for infants aged 6 months or less. In embodiments of the invention, the composition is administered during the first 6 months of life, the first 1 year of life, the first 3 years of life, the first 7 years of life, and/or during the recovery period from an illness or developmental disability.

Nutritional composition

The composition according to the invention is preferably an artificial nutritional composition. The composition of the invention may be, for example, an initial infant formula, an infant formula, an infant food, a cereal-based infant formula, a formula for feeding infants, or an infant formula for children from 1 to 3 years old, and said composition is preferably an initial infant formula. The composition according to the invention may also be intended for use before and/or during a weaning period. In one embodiment, the nutritional composition may be a complete nutritional composition or supplement for aged, elderly, or vulnerable individuals.

The composition according to the invention may be a complete composition providing 100% or most of the nutritional requirements of the target population (e.g. in terms of caloric requirements; or in terms of vitamin or mineral requirements, or in terms of protein requirements , lipids or carbohydrates). In an alternative embodiment, the composition of the invention may be a supplement that is a supplement to the consumer's normal diet. However, in this case, the dosage and total consumption of the composition is adjusted to provide the claimed short-term memory benefit (eg, in proportion to caloric load and the subject's caloric requirements).

The use of the composition of the invention may cover cases where the composition is an additive, preferably provided in unit doses. In one embodiment, the composition is a human breastfeeding supplement.

The composition may be in the form of a powder composition, for example intended to be diluted with water, or mixed with milk (eg human breast milk), or consumed as a powder. In one embodiment, the composition of the invention is in liquid form; either ready-to-eat or intended to be diluted in water or mixed with milk (eg human breast milk).

The composition according to the present invention may also contain a protein source, preferably in an amount of less than 2.5 g per 100 kcal or less than 2.0 g per 100 kcal, even more preferably in an amount of less than 1.8 g per 100 kcal. In one embodiment, the protein content is less than 1.6 g/100 kcal. The type of protein is not considered critical to the present invention, provided that the minimum requirements for the content of essential amino acids are met and satisfactory growth is ensured. Thus, protein sources based on whey, casein and mixtures thereof, as well as protein sources based on soy, can be used. With respect to whey proteins, the protein source may be based on acid whey or sweet whey, or mixtures thereof, and may include alpha-lactalbumin and beta-lactoglobulin in any desired proportions.

The composition in accordance with the present invention, essentially, contains a source of carbohydrates. This is especially preferred when the nutritional composition of the invention is an infant formula. In this case, any carbohydrate source commonly found in infant formulas can be used, such as lactose, sucrose, maltodextrin, starch, and mixtures thereof, although lactose is the preferred carbohydrate source.

The composition in accordance with the present invention, essentially, contains a source of lipids. This is especially important if the nutritional composition of the invention is an infant formula. In this case, the lipid source can be any lipid or fat that is suitable for use in infant formulas. Preferred fat sources include palm olein, high oleic sunflower oil, and high oleic safflower oil. In addition, the essential fatty acids linolenic acid and α-linolenic acid, as well as small amounts of oils containing large amounts of arachidonic acid and docosahexaenoic acid in the finished form, such as fish oils or microbial oils, can be added. The fat source preferably has an n-6 to n-3 fatty acid ratio of about 5:1 to about 15:1; for example, from about 8:1 to about 10:1.

The composition of the invention also preferably contains all the vitamins and minerals known to be essential in the daily diet in nutritionally significant amounts. There are minimum requirements for certain vitamins and minerals. Examples of minerals, vitamins and other nutrients that are optionally present in the composition of the invention include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine and L-carnitine. Minerals are usually added in the form of salts. The presence and amounts of specific minerals and other vitamins may vary depending on the intended population.

If necessary, the composition of the invention may contain emulsifiers and stabilizers such as soy, lecithin, citric acid mono- and diglycerides, and the like.

The composition of the invention may also contain other substances that may have a beneficial effect, such as lactoferrin, nucleotides, nucleosides, and the like.

In one embodiment, the composition of the invention, especially in the form of infant formula, contains from about 1.8 to about 2.2 g of total protein per 100 kcal, for example, from about 1.8 to about 2.1 g, or from about 1. 9 to about 2.1 g of protein per 100 kcal, optionally from about 0.3 to about 0.4 g/100 kcal of protein is alpha-lactalbumin. The infant formula and formula for feeding infants of the present invention may be in the form of a ready-to-drink liquid, or may be a liquid concentrate or powder formula that can be reconstituted into a ready-to-drink liquid by adding some water to produce a result, and the formula for feeding children, which is the subject of the present invention, includes all ingredients required by US or EU law, including, without limitation, certain vitamins, minerals and essential amino acids. It may also include nucleotides such as CMP, UMP, AMP, GMP and IMP, lutein, zeaxanthin and other ingredients known in the art.

In one embodiment, the nutritional composition is a pet food (eg, for dogs or cats or young dogs or young cats).

Effect(s) and application of the composition of the invention

The present invention relates to improving and/or activating and/or improving a subject's short-term memory function. This may include enhancement and/or activation and/or enhancement of working memory and/or declarative memory. As a result of improving the function of short-term memory, the composition of the invention can also positively influence the ability of the subject to acquire and process new knowledge (learning skills).

One embodiment of the invention also improves long-term memory function. One embodiment selectively improves short term memory without improving long term memory.

Without wishing to be bound by theory, it is believed that in one aspect of the invention, such improvement, improvement and/or activation is due to the effect of the oligosaccharide of the invention on the intestinal microbiota. BMO, 2FL, or LNnT are only slightly degraded in the small intestine, so most of them become available for fermentation by the microbiota in the colon. Indeed, it has been shown that BMO or 2FL or LNnT activate the growth of intestinal bifidobacteria, which is reflected in the increase in the content of bacteria of this family in the stool of infants receiving mixtures with the addition of oligosaccharides (with BMO or 2FL and/or LNnT). Briefly, the present nutritional composition can be used to improve memory function through its effects on the gut microbiota and the gut-brain axis, which have been shown to affect many aspects of brain function. In addition, it can be envisaged that the improvement in memory function may be associated with an increase in the concentration of sialic acid (Neu5Ac) in the brain of the specified individual, mediated by various metabolic pathways that are affected by the intestinal microbiota.

In another aspect, the effect of the invention may be associated with an improvement in neuronal plasticity in the subject's brain and/or with an improvement in neuronal development, neurogenesis, axon sprouting, and/or maturation of the subject's brain (without being limited by any theory).

Declarative memory, working memory, and more specifically, short-term memory are important aspects of brain function and development, and are critical to a subject's overall cognitive development. It affects, in particular, the processing, classification and control of external signals, the possibility of learning, as well as spatial orientation.

In one embodiment of the present invention, the declarative memory is a recognition memory. Recognizing memory is extremely important for social and cognitive development, especially in young subjects.

In one embodiment, the present invention relates to the use of (or is defined by) a nutritional composition containing a human milk oligosaccharide (e.g., 2FL and/or LNnT) and/or a fructose oligosaccharide (FOS) to improve, improve or activate declarative memory, short-term memory and/or working memory in a mammal.

In one embodiment, the present invention relates to the use of (or is defined by) a nutritional composition described herein, wherein said composition is an infant formula, a formula for feeding infants, a breast milk fortifier, an infant formula for children from 1 to 3 years old, or a food for pets.

In one embodiment, the present invention relates to (or is defined by) human milk oligosaccharide (e.g., 2FL and/or LNnT) for use in improving, enhancing, or activating short-term memory in a mammal, said mammal being an infant or young child.

In one embodiment, the present invention relates to (or is defined by) a nutritional composition containing a human milk oligosaccharide (e.g., 2FL and/or LNnT) for use in improving, enhancing, or activating declarative memory, working memory, and/or working memory in a young adult. human, preferably in an infant or young child. The nutritional composition may also contain one or more of the optional ingredients described herein.

In one embodiment, the present invention relates to (or is defined by) the use of a human milk oligosaccharide (e.g., 2FL and/or LNnT), in the manufacture of a nutritional composition to improve, enhance or activate declarative memory, working memory and/or working memory in an infant or young child.

Dosage

Fructooligosaccharide/FOS/OF. If available, the nutritional composition of the present invention may contain 0.1 to 20 g of oligofructose (OF) per 100 g of composition based on dry weight, for example 1 to 6 g or 3 to 5 g of oligofructose (OF) per 100 g of composition in terms of dry weight.

One embodiment of the invention nutritional composition contains the amount of fructose oligosaccharide in the following ranges or value:

0.1 to 20 g/l, or 0.5 to 10 g/l, or 1 to 8 g/l, or 2 to 6 g/l, or 1.5 g/l, or 3 g /l, or 5 g/l of the nutritional composition when the composition is in ready-to-use liquid form, or

0.1 to 20 g/l, or 0.5 to 10 g/l, or 1 to 8 g/l, or 2 to 6 g/l, or 1.5 g/l, or 3 g /l, or 5 g/l (liquid reconstituted form), when the composition is in powder form and is intended to be diluted to obtain a reconstituted liquid form, or

the same as above multiplied by 2, 5, 10, 20, 50 or 100 when the nutritional composition is in the form of a concentrated composition intended to be diluted (respectively 2, 5, 10, 20, 50 or 100 times) in water or human breast milk, or intended for direct use in concentrated form, or

0.4 g to 15 g/100 g of powdered nutritional composition, or 0.8 to 10 g/100 g, or 1 to 6 g/100 g, or 2 to 5 g/100 g, or 2 .1 to 4 g/100 g or 1.2 g/100 g or 2.3 g/100 g or 3.8 g/100 g or 4 g/100 g or 6 g/100 g powdered nutrient compositions if the nutritional composition is in the form of a dry powder.

In one embodiment, the OF content may be from 0.07 g to 3 g/100 kcal of the powdered nutritional composition, or from 0.1 to 2 g/100 kcal, or from 0.4 to 1.5 g/100 kcal, or 0.45 to 1 g/100 kcal or 0.45 to 0.75 g/100 kcal or 0.3 g/100 kcal or 0.4 g/100 kcal or 0.5 g/100 kcal, or 0.75 g/100 kcal, or 1 g/100 kcal of powdered nutritional composition if the nutritional composition is in the form of a dry powder.

2FL. The nutritional composition of the present invention may contain from 0.02 to 10 g of 2FL per 100 g of composition, based on dry weight, for example, from 0.2 to 0.5 g or from 0.3 to 5 g of 2FL per 100 g of composition, calculated on dry weight.

In one embodiment of the invention, the nutritional composition contains an amount of 2FL in the following ranges or values:

0.05 to 20 g/l or 0.1 to 5 g/l or 0.2 to 3 g/l or 0.1 to 2 g/l or 0.25 g/l l to 1 g/l, or 0.25 g/l, or 1 g/l of the nutritional composition when the composition is in ready-to-use liquid form, or

0.05 to 20 g/l or 0.1 to 5 g/l or 0.2 to 3 g/l or 0.1 to 2 g/l or 0.25 g/l l to 1 g/l, or 0.25 g/l, or 1 g/l (liquid reconstituted form), when the composition is in powder form and is intended to be diluted to obtain a reconstituted liquid form, or

the same as above multiplied by 2, 5, 10, 20, 50 or 100 when the nutritional composition is in the form of a concentrated composition intended to be diluted (respectively 2, 5, 10, 20, 50 or 100 times) in water or human breast milk, or intended for direct use in concentrated form, or

0.04 g to 1.5 g/100 g of powdered nutritional composition, or 0.08 to 1.2 g/100 g, or 0.1 to 1 g/100 g, or 0.2 to 0 .8 g/100 g or 0.2 g/100 g or 0.4 g/100 g or 0.8 g/100 g or 1 g/100 g or 1 g/100 g powder nutritional composition if the nutritional composition is in the form of a dry powder.

In one embodiment, the 2FL content may be from 0.01 g to 0.3 g, or 3 g/100 kcal of the powdered nutritional composition, or from 0.02 to 0.2 g/100 kcal, or from 0.04 to 0. 15 g/100 kcal, or 0.02 g/100 kcal, or 0.04 g/100 kcal, or 0.07 g/100 kcal, or 0.15 g/100 kcal, or 0.3, or 1, or 2 g/100 kcal of the powdered nutritional composition if the nutritional composition is in the form of a dry powder.

LNnT. The nutritional composition of the present invention may contain from 0.01 to 5 g LNnT per 100 g of composition based on dry weight, for example from 0.1 to 0.25 g or from 0.15 to 0.5 g LNnT per 100 g of composition in terms of dry weight.

In one embodiment of the invention, the nutritional composition contains an amount of LNnT in the following ranges or values:

0.02 to 10 g/l, or 0.05 to 2.5 g/l, or 0.1 to 1.5 g/l, or 0.05 to 1 g/l, or 0 12 g/l to 0.5 g/l, or 0.12 g/l, or 0.5 g/l, or 1 g/l of the nutritional composition when the composition is in ready-to-use liquid form, or

0.02 to 10 g/l, or 0.05 to 2.5 g/l, or 0.1 to 1.5 g/l, or 0.05 to 1 g/l, or 0 ,12 g/l to 0.5 g/l, or 0.12 g/l, or 0.5 g/l, or 1 g/l (liquid diluted form) when the composition is in powder form and intended to be diluted to form a diluted liquid form, or

the same as above multiplied by 2, 5, 10, 20, 50 or 100 when the nutritional composition is in the form of a concentrated composition intended to be diluted (respectively 2, 5, 10, 20, 50 or 100 times) in water or human breast milk, or intended for direct use in concentrated form, or

from 0.02 g to 0.75 g/100 g of powdered nutritional composition, or from 0.04 to 0.6 g/100 g, or from 0.05 to 0.5 g/100 g, or from 0.1 up to 0.4 g/100 g, or 0.1 g/100 g, or 0.2 g/100 g, or 0.25 g/100 g, or 0.5 g/100 g, or 1 g/100 g, or 3 g/100 g of powdered nutritional composition if the nutritional composition is in the form of a dry powder.

In one embodiment, the content of LNnT may be from 0.01 g to 0.3 or 1 or 3 g/100 kcal of the powdered nutritional composition, or from 0.02 to 0.2 g/100 kcal, or from 0.04 to 0 .15 g/100 kcal, or 0.02 g/100 kcal, or 0.04 g/100 kcal, or 0.07 g/100 kcal, or 0.15 g/100 kcal, or 0.3 g/100 kcal of the powdered nutritional composition if the nutritional composition is in the form of a dry powder.

The composition of the invention, in particular in the form of infant formula, may contain at least about 0.4 g of oligofructose per 100 kcal. In some embodiments, it contains about 0.4 to about 0.9 g, about 0.4 to about 0.7 g, about 0.4 to about 0.5 g, about 0.7 to about 0 .8 g or from about 0.7 to about 0.9 g of oligofructose per 100 kcal. The oligofructose has a degree of polymerization from 2 to 10. In one embodiment, at least 90% of the oligofructose has a degree of polymerization from 2 to 8.

In one embodiment, the nutritional composition contains 2 FL and LNnT, preferably in an amount of 1 g/l 2FL and 0.5 g/l LNnT, or 0.5 g/l 2FL and 0.25 g/l LNnT, or from 0, 25 g/l to 2 g/l 2FL and 0.1 to 1 g/l LNnT.

Method for preparing a nutritional composition

The nutritional composition can be prepared by any suitable method known in the art. For example, a commercial infant formula or formula for feeding infants may serve as the base composition to which the required amount of oligosaccharides (eg 2FL, LNnT or others) is added, preferably in dry form. In an alternative embodiment, the oligosaccharide can be added as a dry ingredient or liquid ingredient to a liquid premix that will serve as the basis for the production of the nutritional composition of the invention. The liquid mixture can then be dried by any conventional means.

For example, it can be prepared by mixing together a protein source, a carbohydrate source (other than the oligosaccharide combination of the present invention) and a fat source in appropriate proportions. At this stage, emulsifiers can be included if they are used. Vitamins and minerals can be added at this stage, but they are usually added later to avoid thermal decomposition. Any lipophilic vitamins, emulsifiers, etc. may be dissolved in the fat source prior to mixing. Water, preferably reverse osmosis purified water, may then be admixed to subsequently form a liquid mixture. The temperature of the water conveniently ranges from about 50 [deg.] C. to about 80 [deg.] C. to facilitate dispersion of the ingredients. Commercially available thinners can be used to form the liquid mixture. 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL) are added at this stage if the final product is to be in liquid form. If the end product is to be a powder, 3'-sialyllactose (3'-SL) and 6'-sialyllactose (6'-SL) can also be added at this stage if necessary. The liquid mixture is then homogenized, for example in two stages.

Subsequently, the liquid mixture can be thermally treated to reduce the bacterial load by rapidly heating the liquid mixture, for example, to a temperature in the range from about 80 [deg.] C to about 150 [deg.] C for a time from about 5 seconds to about 5 minutes. The heating can be carried out by steam injection using an autoclave or a heat exchanger such as a plate heat exchanger. The liquid mixture can then be cooled to a temperature of from about 60 [deg.] C to about 85 [deg.] C, for example, by flash cooling. The liquid mixture can then be homogenized again, for example in two stages: under pressure in the range of about 10 MPa to about 30 MPa in the first stage and from about 2 MPa to about 10 MPa in the second stage. The homogenized mixture can then be further cooled to add any heat sensitive components such as vitamins and minerals. At this stage, it is convenient to adjust the pH and solids content of the homogenized mixture. The homogenized mixture is transferred to a suitable drying apparatus such as a spray dryer or freeze dryer and powdered. The moisture content of the powder should be less than about 5% by weight.

If a liquid composition is preferred, the homogenized mixture may be sterilized and then aseptically filled into suitable containers, or may be first filled into containers and then autoclaved.

Specific lipids

The composition of the invention may contain selected lipids that have certain effects.

Such lipids may in particular include DHA, ARA, linoleic acid or sphingomyelin, preferably in an amount acceptable to provide real brain health benefits and within the general regulatory requirements for this type of product (for example, WHO recommendations for infant formula; CODEX or European Infant Formula Directives).

In some embodiments, the implementation of the composition of the invention contains a relatively high level of sn-2-palmitate or sphingomyelin. They have been associated with optimal performance and development of the brain and may provide a synergistic effect with the required compounds of the composition of the invention.

Although feeding a high sn-2-palmitate formula (in the absence of OF) to an infant helps to promote the growth of bifidobacteria in the colon, the combination of high sn-2-palmitate with oligofructose is believed to result in a significantly higher growth of bifidobacteria in the colon in formula-fed infants. In addition, a significant reduction in the number of potentially pathogenic bacteria can be achieved. Feeding an infant formula containing sn-2-palmitate and about 3 to about 5 g/L, or about 0.4 to about 0.7 g/100 kcal of oligofructose, has been found to be more beneficial than feeding an infant. the same mixture without oligofructose. Without being limited by any theory, such a synergistic effect between OF and sn-2-palmitate may also activate short-term memory, in particular (but most likely not only) through the effect on the subject's microbiome and bifidobacteria population. The term "high content of sn-2-palmitate" should be understood as ingredients having a high percentage of fatty acids (preferably more than 33% fatty acids) in the form of palmitate in the sn-2-position of triglycerides. Such ingredients are commercially available under the trade name BETAPOL® (Loders Croklaan, Vormerveer, The Netherlands) or INFAT® (Advanced Lipids AB, Karlshamn, Sweden, a joint venture between AAK B.V. (Zaandijk, The Netherlands) and Enzymotec Inc., . Morristown, New Jersey, USA)

Recent clinical studies in infants have shown that formulas containing at least one omega-6 fatty acid and at least one omega-3 fatty acid in a ratio of about 6 to about 1 increase DHA accumulation in red blood cells and blood plasma. A balanced ratio of about 6:1 of omega-6 fatty acids to omega-3 fatty acids may also provide long-term health benefits, including optimal brain and nervous system development. This balance will be achieved by formulating compositions of the present invention with fat sources from vegetable oils that have omega-6 fatty acids in their composition, such as, for example, soybean oil and sunflower oil, and which have omega-3 fatty acids in their composition, such as rapeseed, canola, flaxseed, chia, pearl or walnuts. A unique fat blend with 5 different oils will be applied to provide a modified fat blend.

The following examples are presented to illustrate certain embodiments and features of the present invention, but should not be construed as limiting the scope of the present invention.

Examples

Example 1 Nutritional composition containing 2FL and/or LNnT

A nutritional composition of the invention containing oligofructose (OF) and/or 2FL and/or LNnT is shown in Table 1 below. This composition is for illustrative purposes only. The composition of Table 1 may be an infant formula. In an alternative embodiment, it may be formulated to be suitable for use as formula for formula fed infants.

Table 1

Nutrients Per 100 kcal per liter Energy value (kcal) 100 670 Protein (g) 1.83 12.3 Fat (g) 5.3 35.7 Linoleic acid (g) 0.79 5.3 α-Linolenic acid (mg) 101 675 lactose (g) 11.2 74.7 Minerals (g) 0.37 2.5 Na (mg) 23 150 K (mg) 89 590 Cl (mg) 64 430 Ca (mg) 62 410 P (mg) 31 210 Mg (mg) 7 50 Mn (mcg) 8 50 Se (mcg) 2 13 Vitamin A (µg RE) 105 700 Vitamin D (mcg) 1.5 10 Vitamin E (mg TE) 0.8 5.4 Vitamin K1 (mcg) 8 54 Vitamin C (mg) 10 67 Vitamin B1 (mg) 0.07 0.47 Vitamin B2 (mg) 0.15 1.0 Niacin (mg) 1 6.7 Vitamin B6 (mg) 0.075 0.50 Folic acid (mcg) 9 60 Pantothenic acid (mg) 0.45 3 Vitamin B12 (µg) 0.3 2 Biotin (mcg) 2.2 15 Choline (mg) 10 67 Fe (mg) 1.2 8 I (µg) 15 100 Cu (mg) 0.06 0.4 Zn (mg) 0.75 5 2FL (g)
and/or
LNnT (g) 0.04 or 0.15
0.01 or 0.07 0.25 or 1
0.1 or 0.5 Oligofructose (OF) (g) (optional) 0.45 or 0.75 3 or 5 3'-SL (g) (optional) 0.22 1.46 6'-SL (g) (optional) 0.05 0.33 3'-SL : 6'-SL ratio (optional) 4.4:1 4.4:1 BMO (g) (optional) 0.4 or 2 3 or 15

Example 2 Infant formula containing HMO (2 FL and/or LNnT), optional BMO and/or oligofructose (OF)

A nutritional composition of the invention containing oligofructose (OF) and optionally 2FL and/or LNnT is shown in Table 2 below. This composition is for illustrative purposes only. The composition shown in Table 2 is an infant formula. In an alternative embodiment, it may be formulated to be suitable for use as formula for formula fed infants.

Another example is based on commercial Nan infant formulas and/or lactogenic infant formulas (manufactured by Nestlé, Switzerland) to which the specific oligosaccharides of the invention are added in the amount indicated below.

table 2

Nutrient Per 100 kcal per liter Energy value (kcal) 100 670 Protein (g) 1.83 12.3 Fat (g) 5.3 35.7 Linoleic acid (g) 0.79 5.3 α-Linolenic acid (mg) 101 675 lactose (g) 11.2 74.7 Prebiotic (70% OF, 30% inulin) (g)
(e.g. Beneo-Orafti® P95 and Inulin Beneo-Orafti®)
(not necessary) 0.64 4.3 2FL (g)
and/or
LNnT (mg) 0.3
thirty 2
200 Minerals (g) 0.37 2.5 Na (mg) 23 150 K (mg) 89 590 Cl (mg) 64 430 Ca (mg) 62 410 P (mg) 31 210 Mg (mg) 7 50 Mn (mcg) 8 50 Se (mcg) 2 13 Vitamin A (µg RE) 105 700 Vitamin D (mcg) 1.5 10 Vitamin E (mg TE) 0.8 5.4 Vitamin K1 (mcg) 8 54 Vitamin C (mg) 10 67 Vitamin B1 (mg) 0.07 0.47 Vitamin B2 (mg) 0.15 1.0 Niacin (mg) 1 6.7 Vitamin B6 (mg) 0.075 0.50 Folic acid (mcg) 9 60 Pantothenic acid (mg) 0.45 3 Vitamin B12 (µg) 0.3 2 Biotin (mcg) 2.2 15 Choline (mg) 10 67 Fe (mg) 1.2 8 I (µg) 15 100 Cu (mg) 0.06 0.4 Zn (mg) 0.75 5 3'-sialyllactose (mg) (optional) thirty 200 6'-sialyllactose (mg) (optional) 6 40 BMO (g) (optional) 0.9 6

Experimental data

In a study conducted on piglets, the effect of various oligosaccharides on recognition memory (human milk oligosaccharides, cow's milk oligosaccharides) was studied. Forty-eight male piglets (12 per treatment group) received control (CON, Purina ProNurse animal milk replacer), human milk oligosaccharide (HMO) (CON + 1.5 g/l (consisting of 1.0 g/l 2'FL + 0.5 g/l LNnT)) or cow's milk oligosaccharides (BMO) (6.5 g/l BMOS) or HMO + BMOS (1.0 g/l 2'FL + 0.5 g/l LNnT + 6.5 g/l BMOS). All diets contained an 8.0 g/L formulation change reserve that was reserved for the addition of the test diet components. Piglets were fed with 285 ml and 325 ml of the diluted ration per kg of body weight on days 2-6 after birth (PND 2-6) and days 7-33 after birth (PND 7-33), respectively. Piglets were tested for recognition memory between PND 22 and 31 and had an MRI scan on the 32nd or 33rd day.

The BMOS mixture used in infant formulas was derived from cow's milk whey. Briefly, the bovine serum ultrafiltrate, including oligosaccharides such as 3'- and 6'-sialyllactose and GOS, was demineralized by a combination of electrodialysis and ion exchange as described in the art.

Behavioral Testing

New Object Recognition (NOR). Recognizing memory was assessed using NOR, a task that depends on the function of the perirhinal cortex. The trial consisted of an habituation phase, a reference phase, and a trial phase. During the habituation stage, piglets were placed in an empty test arena for 10 minutes each day for two days until the reference phase. In the reference phase, a pig was placed in an arena containing two identical objects and given 5 minutes to study. After a delay of 1 hour or 2 days (48 hours), the piglet was returned to the arena for the testing phase. In the test phase, the pig was placed in an arena containing one object from the reference stage as well as a new object and given 5 minutes to study. Between tests, objects were removed, immersed in hot water with detergent and wiped with a towel to reduce odor, and the arena was sprayed with water and cleaned to remove urine and feces. The selected objects had a certain set of characteristics (i.e. color, texture, shape and size), but the new objects and standards differed only in shape and size. For testing, we used only objects for which the preference was previously zero.

The order of tasks between repetitions was balanced. Habituation tasks were started at 22 days of age, and reference tasks in the NOR task were started at 25 days of age. Measured the time spent on the study of objects, and the distance of movement, using a combination of automated procedures using the Ethovision system and manual tracking. Exploratory behavior was broken down into 20 behavioral endpoints to assess the effect of diet on behavioral performance. The recognition ratio was used to measure recognition memory (time spent learning a new object)/(time spent learning a new object + time spent learning a known object).

Statistical analysis

All data from this study were subjected to analysis of variance (ANOVA) using the mixed procedure MIXED from SAS 9.4 (SAS Inst. Inc., Cary, NC, USA). All statistical models used included repetition as a random effect, and data collected at one time point (during habituation and reference stages) were analyzed by 2-way ANOVA, and 2) data collected from one animal by more than one case (eg, multiple tests, etc.) were analyzed by a 3-way repeated measures analysis of variance. Post hoc comparisons were made for significant interaction effects using Tukey's adjusted test. For some behavioral outcomes (Table 2), a one-sample t-test was obtained. In all cases, statistical significance was defined as P<0.05 and trend was defined as P<0.10.

results

Recognition coefficient

The effect of diets on the recognition rate is shown in Table 3 and in FIG. 1.

results

The use of HMO alone resulted in a statistically significant increase in recognition memory with a 1-hour intertest interval, but unexpectedly did not improve recognition memory over a 2-hour intertest interval. This indicates a clear positive effect of HMO (2' FL + LNnT) on short-term recognition memory. The use of HMO + BMO leads to a numerical increase in recognition memory with a 1-hour intertest interval and a statistically significant increase in recognition memory with a 24-hour intertest interval.

In other words, the inventors observed a statistically significant increase in recognition memory, as measured by recognition quotient, in the group treated with HMO (2'FL + LNnT) with 1 hour delay (short-term memory) compared to CON. This effect was observed against the background of no change in long-term recognition memory (2-day delay). Only BMO (cow's milk oligosaccharides), acting as a negative control, had no specific effect on short-term memory. An effect on short-term memory (1 hour) also seemed to be present in the HMO + BMO group. In addition, a statistically significant increase in recognition memory was also observed in the group treated with HMO (2' FL and LNnT) + BMOS with a 2-day delay (long-term memory).

These results show the effect of nutritional intervention with BMO on short-term memory and BMO + BMO for both short-term and long-term memory.

Table 3. Effect of oligosaccharide supplementation on recognition memory during a pilot study of a novel object recognition task

Delay Diet Quantity Recognition coefficient Standard osh. middle P-value ³ 1 hour Control 10 0.515 0.055 0.397 OGM 10 0.619 0.059 0.038 BMO 10 0.469 0.071 0.663 OGM + BMO 10 0.593 0.064 0.090 48 hours Control 10 0.520 0.085 0.372 OGM eleven 0.492 0.081 0.540 BMO 8 0.548 0.091 0.309 OGM + BMO 10 0.598 0.052 0.045

³ P is the value from the one-tailed t-test for a recognition rate greater than 0.50.

Claims (7)

1. The use of a nutritional composition containing breast milk oligosaccharide (HMO) to improve or activate declarative memory in an infant or young child, and the composition contains breast milk oligosaccharides 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose ( LNnT), and the composition additionally contains cow's milk oligosaccharides 3'-sialyllactose and 6'-sialyllactose and galactooligosaccharide (GOS). 2. The use of a nutritional composition containing a breast milk oligosaccharide (HMO) according to claim 1 in an infant or young child, wherein said declarative memory is an identifying memory. 3. The use of a nutritional composition containing breast milk oligosaccharide (HMO) according to paragraph 1 or 2 in an infant or young child, and HMO is present in an amount of from 0.1 to 10 g/l, or from 0.3 to 5 g /l, or from 0.5 to 1 g/l, or 0.25, or 0.5, or 1, or 1.5, or 2 g/l. 4. Use of a nutritional composition according to any of the preceding claims, which further comprises a prebiotic, said prebiotic being selected from inulin, xylooligosaccharides, polydextrose, fructose oligosaccharide (FOS) or oligofructose (OF), or any combination thereof. 5. Use of a nutritional composition according to any one of the preceding claims, wherein the ratio of 3'-sialyllactose to 6'-sialyllactose is in the range of 5:1 to 1:2. 6. Use of a nutritional composition according to any of the preceding claims, further comprising a probiotic, wherein the probiotic is a probiotic bacterial strain selected from the list consisting of Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus lactis, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus bulgari, Lactococcus lactis, Lactococcus diacetylactis, Lactococcus cremoris, Streptococcus salivarius, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis или Bifidobacterium adolescentis или любой их mixtures. 7. The use of a nutritional composition according to any one of the preceding claims, said composition being an infant formula, a formula for feeding infants, a breast milk fortifier or an infant formula for children from 1 to 3 years of age.

Images