RU2776182C2 - Compositions with specific oligosaccharides for prevention of obesity later in life or related comorbidities by means of increasing scfa production in colon and/or by means of increasing glp-1 secretion - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: present invention relates to the use of an artificial nutritional composition containing a mixture of oligosaccharides for the reduction and/or avoidance of excessive accumulation of adipose tissue mass in an infant or in a young child, and/or for the prevention of a health disorder later in life, related to excessive accumulation of adipose tissue mass in an infant or a young child, selected from a list consisting of insulin resistance, glucose tolerance disorder, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gallbladder disease, cardiovascular disease and metabolic syndrome, by means of increasing SCFA production in the colon and/or by means of increasing GLP-1 secretion in the specified infant or young child. In this case, the specified mixture of oligosaccharides contains at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide, and at least one sialylated oligosaccharide.

EFFECT: present invention provides for an increase in an animal model in the production of short-chain fatty acids (SCFA) in the colon, such as propionate, butyrate, valerate, acetate, due to the use of a mixture of cow milk oligosaccharides and increases the secretion of glucagon-like peptide 1 (GLP-1) in the in vitro system using human enteroendocrine cells.

14 cl, 4 dwg, 3 tbl, 3 ex

Description

Scope of the invention

The present invention relates to nutritional compositions, such as infant formula containing a mixture of oligosaccharides, which is specifically designed to avoid excessive accumulation of adipose tissue mass and to prevent associated health disorders later in life in infants or young children, similar to obesity in later life, by increasing colonic SCFA production and/or GLP-1 secretion in said infants or young children.

Prerequisites for the creation of the invention

Scientific evidence suggests that infancy may be a critical period in the development and programming of future health conditions, including obesity later in life or future associated comorbidities (including metabolic disorders).

Overweight and obesity are defined as the abnormal or excessive accumulation of fat that can be detrimental to health. Body mass index (BMI) is a simple weight-for-height measure that is widely used to classify overweight and obesity in adults. It is defined as the subject's mass in kilograms divided by the square of their height in meters (kg/ ^m2 ). The WHO definition is as follows: A BMI greater than or equal to 25 indicates overweight; A BMI greater than or equal to 30 indicates obesity. Over the past 30 years worldwide, the prevalence of obesity and overweight in adults, children and adolescents has risen dramatically and continues to rise. Globally, the incidence of obesity has more than doubled since 1980, according to WHO. It has become a global health problem as it is associated with reduced life expectancy, altered quality of life and in doing so it is responsible for further health conditions. Globally, overweight and obesity are associated with more deaths than underweight. Childhood obesity is indeed associated with a higher likelihood of obesity, premature death and disability in adulthood.

But in addition to increased future risks, obese children experience breathing difficulties, increased risk of fractures, hypertension, early markers of cardiovascular disease, insulin resistance, and physiological effects. Elevated BMI is a major risk factor for non-communicable diseases such as cardiovascular disease (predominantly heart disease and stroke), which was the leading cause of death in 2012; diabetes; musculoskeletal disorders (especially osteoarthritis, a highly incapacitating degenerative joint disease); even some types of malignant tumors (endometrium, breast and colon).

Insulin resistance can lead to type 2 diabetes. Type 2 diabetes is a chronic disease associated with abnormally high levels of sugar-glucose in the blood. In type 2 diabetes, the amount of insulin is generally sufficient, but the cells it is supposed to act on do not show normal sensitivity to its action. Signs and symptoms include increased urine output and decreased appetite, as well as fatigue. The main complications of diabetes include dangerously high blood sugar levels, abnormally low blood sugar levels due to diabetes medications, and blood vessel disease that can affect the eyes, kidneys, nerves, and heart.

Metabolic syndrome is a grouping of at least three of the following five medical conditions: abdominal (central) obesity, elevated blood pressure, elevated fasting plasma glucose, high serum triglycerides, and low high-density lipoprotein (HDL) levels . Metabolic syndrome is associated with the risk of developing cardiovascular disease and diabetes. Some studies have shown an estimated prevalence in the US of 34% of the adult population, and prevalence increases with age.

Mother's milk is recommended for all babies for various reasons. Breastfeeding has been specifically reported to be particularly beneficial compared to formula feeding in the prevention of health problems later in life, such as obesity later in life (Owen et al, Effect of Infant Feeding on the Risk of Obesity Across the Life Course: A Quantitative Review of Published Evidence, 2005) and type 2 diabetes in later life (Owen et al, Does breastfeeding influence the risk of type 2 diabetes in later life? A quantitative analysis of published evidence, 2006). It has been widely reported that growth patterns differ between breastfed and formula fed infants. Indeed, breastfed infants have lower weight gain and lower body fat mass during the first year of life compared to formula fed infants. In addition, a breastfed infant has a different gut microbiota profile compared to formula fed infants. In general, these factors influence the development of an infant's physiology, including metabolism, immunity, and overall growth.

However, in some cases, breastfeeding is insufficient or unfavorable for medical reasons, or the mother refuses to breastfeed. Infant formulas have been developed for these situations. Fortifiers have also been developed to fortify mother's milk or infant formula with certain ingredients.

Short-chain fatty acids (SCFAs) are produced, in part, by microbial fermentation of dietary fiber in the colon. SCFAs, especially butyrate and propionate, have been shown to protect against obesity, insulin resistance, and are involved in adipogenesis, dietary intake, and the prevention of non-alcoholic fatty liver disease or conditions associated with cardiac metabolism, such as atherosclerosis and inflammation (Arora et al., Propionate: Anti-obesity and satiety enhancing factor?, 2011; Lin et al., Butyrate and Propionate Protect against Diet-Induced Obesity and Regulate Gut Hormones via Free Fatty Acid Receptor 3-Independent Mechanisms, 2012; Chambers et al. , "Effects of targeted delivery of propionate to the human colon on appetite regulation, body weight maintenance and adiposity in overweight adults", 2014; Canfora et al, "Short-chain fatty acids in control of body weight and insulin sensitivity", Nat Rev Endocrinol 11, 577–591, 2015; Aguilar et al, "Butyrate impairs atherogenesis by reducing plaque inflammation and vulnerability and decreasing NFκB activation", 2015; Endo et al, "Butyrate-p roducing probiotics reduce nonalcoholic fatty liver disease progression in rats: new insight into the probiotics for the gut-liver axis”, 2013).

SCFAs like butyrate and propionate have also been shown to significantly stimulate the release of gut hormones like GLP-1 (Chambers et al., 10 December 2014; Hua V. Lin et al, April 2012).

Glucagon-like peptide 1 (GLP-1 or GLP1) is an incretin secreted by intestinal L cells during nutrient uptake. Among its main effects, it improves glucose clearance by stimulating insulin secretion in pancreatic beta cells as well as simultaneously inhibiting glucagon synthesis and secretion (Kreymann B et al. 1987, PMID:2890903; Nathan DM et al. 1992, PMID:1547685 ; Nauck MA et al. 1993, PMID: 8423228; Ritzel R et al. 2001, PMID: 11289042). Therefore, it can carry out the prevention of type II diabetes.

GLP-1 has also been shown to slow gastric emptying (Little TJ et al. 2006, PMID: 16492694; Nauck MA et al. 1997, PMID: 9374685), reduce appetite and food intake in both healthy and obese (Pratley et al. 2008; Orskov et al. 1989; Davis HR et al. 1998; PMID:9545022; Domon-Dell et al. 2002; Drucker 2002; Schusdziarra V et al. 2008, PMID:18281111; Punjabi M et al., 2014; PMID: 24601880).

GLP-1 has also been shown to reduce body weight/BMI (Zaccardi F. et al. 2016, PMID:26642233; Kelly AS et al. 2013, PMID:23380890; Kelly AS et al. 2012, PMID:22076596).

GLP-1 has also been described to provide some beneficial effects on the cardiovascular system (Bose et al. 2005, PMID:15616022; Sokos GG et al. 2006, PMID:17174230).

Increased colonic SCFA production and/or GLP-1 secretion is therefore an attractive target for the prevention of health disorders later in life, especially those due to or associated with fat accumulation.

However, orally administered SCFAs are unpalatable and rapidly absorbed in the small intestine. For this reason, specific delivery systems have been developed for some studies to target the release of propionate in the proximal colon. In Chambers et al, scientists have developed a specific carrier molecule in which propionate is chemically ester-linked to inulin, a natural polymer composed predominantly of fructose. This inulin-propionate ester was chemically synthesized. Most of the propionate chemically bound to inulin should only be released when the inulin polymer is fermented by the colonic microbiota, thus allowing targeted delivery to the colon. However, this type of media has some disadvantages. This type of chemically synthesized material may face regulatory issues when used in formulations intended for infants or young children. This study was indeed intended for adults. Therefore, some more "natural" solutions, such as those with ingredients found in breast milk, will be preferred for administration to infants or young children.

With regard to the secretion of GLP-1, two pharmacological approaches have been developed to increase the activity of GLP-1 or GLP-1-like activity. The first is to reduce the degradation of GLP-1 through inhibition of the enzyme responsible for it (DPP-4i). Additionally, several GLP-1 receptor agonists have been used to increase GLP-1 receptor activation. However, all of these pharmacological approaches are indicated for adults only. For infants and young children, more "natural" solutions will be preferred.

Therefore, alternative solutions more suitable for infants and young children should be developed.

As the composition of human breast milk becomes better understood, it has been suggested that prebiotics be added to infant formula. There are various prebiotic supplemented infant formulas on the market, such as fructooligosaccharides (FOS) and galactooligosaccharides (GOS) mixtures. Prebiotics are non-digestible in the sense that they are not broken down or absorbed in the stomach or small intestine and thus pass unchanged to the large intestine where they are selectively fermented by bacteria. The main effect of prebiotics after fermentation is to selectively stimulate the growth and metabolic activity of certain types of bacteria recognized as beneficial to the well-being and health of the host organism (Roberfroid, M, J. Nutrition, 2007: 37(3): 830S-837S). In addition to the direct effect of the prebiotic on the gastrointestinal flora, it is also known that prebiotics have a beneficial effect on the health of the host organism (for example, anti-carcinogenic effects, improved absorption of minerals and effects on the production of metabolites), which may be due to the indirect effect of prebiotics on the intestinal microflora.

Prebiotics such as oligofructose have been shown to stimulate GLP-1 release in the gut (Cani et al., 2005, Phuwamongkolwiwat et al., 2014). However, the degree of polymerization varies widely from one type to another, and therefore the biological effect induced can vary to a great extent.

Moreover, commercially available mixtures only approximate the mixture of oligosaccharides found in breast milk. More than 120 different oligosaccharide components have been found in breast milk, some of which are still not found at all in animal milk (for example, in cow's milk) or are found only in small quantities. Some classes of human milk oligosaccharides, which are present in cow's milk or colostrum in very small amounts or not at all, are sialylated and fucosylated oligosaccharides. Since cow's milk contains some oligosaccharides that are structurally identical or similar to those found in human milk, cow's milk oligosaccharides in sufficient quantities should have a prebiotic effect or other beneficial properties inherent in human milk oligosaccharides. However, until recently, the low concentration of these oligosaccharides in cow's milk (about 20 times lower than in human milk) has hindered attempts to use cow's milk as a source of oligosaccharides for infant formula.

Additionally, there are several references in the literature relating to the use of cow's milk oligosaccharides for the prevention or treatment of metabolic diseases. Patent application WO2010/003803 describes a nutritional composition for administration to infants in which an oligosaccharide mixture consists of N-acetylated oligosaccharide(s), galactooligosaccharide(s) and sialylated oligosaccharide(s), for reduce the risk of obesity later in life. This composition has been shown to induce a decrease in hepatic triglyceride concentrations with reduced lipogenesis and incorporation of triacylglycerols into hepatic lipoproteins. But none of these prior art documents mentioned or reported the effects of BMO on colonic SCFA production and/or on GLP-1.

There is a clear need to develop new and suitable methods to reduce the risk of later life health conditions associated with excessive accumulation of adipose tissue mass, like obesity or related comorbidities in infants and young children.

There is also a need to provide such health benefits in a manner that is particularly suitable for these young subjects (infants and young children), which method must not include classical pharmaceutical intervention, as infants or young children are particularly vulnerable.

There is a need to provide these health benefits to infants and young children in a manner that does not cause side effects and/or that is easy to carry out and that is acceptable to parents or healthcare professionals.

There is also a need to provide such benefits in a manner that will keep the cost of such provision reasonable and affordable to the majority.

Statement of the Invention

The present inventors have found that a composition containing a specific mixture of bovine milk oligosaccharides (BMO) can increase colonic production of SCFAs like propionate, butyrate, valerate and acetate, and especially butyrate and propionate, in an animal model.

They also found that this specific mixture of bovine milk oligosaccharides (BMO) increased GLP-1 secretion in an in vitro system using human enteroendocrine cells.

This represents new clinical situations in which the goal of preventing excess body fat mass and any associated health problems later in life can be achieved in new ways.

Thus, a composition containing this specific BMO mixture can be advantageously used to prevent in an infant or young child a health disorder later in life associated with excessive accumulation of adipose tissue mass, for example, overweight, obesity, insulin resistance, impaired tolerance to glucose, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gallbladder disease, cardiovascular disease and metabolic syndrome.

In a specific embodiment, the nutritional composition contains from 2.5 to 15.0 wt.% mixture of oligosaccharides.

The mixture of oligosaccharides contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide.

It may contain from 0.1 to 4.0 wt.% N-acetylated(s) oligosaccharide(s), from 92.0 to 99.5 wt.% galactooligosaccharide(s) and from 0.2 to 4 0 wt.% sialylated(s) oligosaccharide(s).

The mixture of oligosaccharides can be obtained from animal milk such as cow's milk, goat's milk or buffalo milk.

Brief description of the figures

In FIG. 1 shows butyrate production in the caecum of mice fed low fiber diets and low fiber diets fortified with 5% of the various fibers tested.

Abbreviations: Positive. counter. = positive control; BMOS = cow's milk oligosaccharides; PDX = polydextrose.

In FIG. 2 shows caecal propionate production in mice fed low fiber diets and low fiber diets fortified with 5% of the various fibers tested.

Abbreviations: Positive. counter. = positive control; BMOS = cow's milk oligosaccharides; PDX = polydextrose.

H fig. 3 shows the ratio of the median content of each SCFA in the fiber-fortified diet divided by the median value for the positive control diet.

Abbreviations: Contr. positive = positive control; BMOS = cow's milk oligosaccharide; PDX = polydextrose.

In FIG. 4 shows GLP-1 secretion in an in vitro system using human NCl-H716 enteroendocrine cells.

Abbreviations: Negative. counter. = negative control; Positive counter. = positive control; BMOS = cow's milk oligosaccharides.

Detailed description of the invention

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

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

The expression "young child" means a child between the ages of one and three, also called a toddler.

"Infant or young child born by caesarean section" means an infant or young child born by caesarean section. This means that the baby or young child was not born through a vaginal birth.

"Infant or young child born by vaginal delivery" means an infant or young child who was born by vaginal delivery and not by caesarean section.

"Premature" or "premature" means an infant or young child who is born prematurely. Essentially, this term refers to an infant or young child born before the 36th week of pregnancy.

The expression "nutritional composition" means the composition that feeds the subject. This nutritional composition will generally be taken orally or intravenously. It may include a lipid or fat source, a carbohydrate source, and/or a protein source. In a specific embodiment, the nutritional composition is a ready-to-use composition, such as a ready-to-use formula.

In a specific embodiment, the composition of the present invention is a hypoallergenic nutritional composition. The term "hypoallergenic nutritional composition" means a nutritional composition that is unlikely to cause allergic reactions.

In a specific embodiment, the nutritional composition of the present invention is an "artificial nutritional composition". The expression "artificial nutritional composition" means a mixture obtained by chemical and/or biological means, which may be chemically identical to the mixture naturally present in the milk of mammals (i.e., a synthetic nutritional composition is not breast milk).

The term "infant formula" as used herein refers to a food product specifically intended for consumption by infants during the first months of life, which by itself satisfies the nutritional needs of this category of persons (Article 2(c) of European Commission Directive 91/ 321/EEC 2006/141/EC of 22 December 2006 on infant formula and infant formula for breastfed babies). It also refers to a nutritional composition intended for infants as defined in the Codex Alimentarius (Codex STAN 72-1981) and the "Special Foods for Infants" standard (including the article "Foods for Special Medical Purposes"). The expression "infant formula" covers both "initial formula" and "formula for formula feeding".

“Follow-up formula” or “pre-fed formula” is given starting at 6 months. It constitutes the main liquid element in the gradually increasing variety of diets for this category of individuals.

The expression "infant food" means a food product specifically intended for use by infants or young children during the first years of life.

The expression "children's cereal-based composition" means a food product specifically intended for use by infants or young children during the first years of life.

The term "fortifier" refers to liquid or solid nutritional compositions that are suitable for mixing with breast milk or infant formula.

The expression "weaning period" means the period during which the diet of an infant or young child replaces mother's milk with another food.

The expressions "aged days/weeks/months/years of life", "days/weeks/months/years after birth" and "days/weeks/months/years from birth" may be used interchangeably.

The expression "in the later life" and "in the afterlife" can be used interchangeably. They refer to effects measured in an individual (infant or young child) after the age of a few weeks, a few months or a few years after birth, for example, after the age of 6 months after birth, for example, after the age of 8 months after birth, for example, after the age 10 months after birth, for example after 1 year of age after birth, for example after 2 years of age, preferably after 4 years of age, more preferably after 5 years of age, even more preferably after 7 years of age or even more, and compared with average observational data for subjects of the same age. Preferably, it refers to an effect observed after at least 1 year of age, or after at least 2, 5, 7, 10 or 15 years of age. Thus, "later in life" may refer to observation during infancy, early childhood, childhood, adolescence, or adulthood. Preferably, it refers to observation in childhood, adolescence or adulthood.

The expressions "accumulation of adipose tissue mass" and "accumulation of fat" can be used interchangeably. The expression "excessive accumulation of adipose tissue mass" refers to an abnormal amount of adipose tissue in the body, for example, an amount that can lead to health problems.

The expressions "reducing excessive accumulation of adipose tissue mass" and "avoiding excessive accumulation of adipose tissue mass" refer to reducing or limiting the amount of fat in the body of an individual in order to achieve a normal or lower adipose tissue mass, for example, in an amount that does not lead to health disorders .

The term "health disorder(s)" encompasses any health condition and/or disease and/or dysfunction that affects an individual's body, including metabolic dysfunctions.

The expressions "prevention of disorder in later life" or "prevention of disorder in later life" can be used interchangeably. They mean avoiding a health disorder/condition occurring later in life and/or reducing the frequency and/or severity of a health disorder later in life. Prevention occurs "later in life", therefore, preferably after the completion of the intervention or treatment (ie, after administration of the nutritional composition of the present invention).

The expression "health disorder in later life associated with excessive accumulation of adipose tissue mass" refers to a health disorder in later life due to (ie, direct relationship) or associated (ie, indirect relationship) with excess fat. It covers overweight, obesity and obesity-related comorbidities.

"Body mass index" or "BMI" is defined as the value obtained by dividing the numerator, which is mass in kilograms, by the denominator, which is the square of height in meters. Alternatively, body mass index can be calculated as the ratio of the weight in pounds in the numerator to the square of the height in inches in the denominator multiplied by 703. "Overweight" is defined for a person as a BMI between 25 and 30. "Obesity" is defined for a person as a BMI over 30.

"Obesity-related comorbidities" include insulin resistance, impaired glucose tolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gallbladder disease, cardiovascular disease, metabolic syndrome and certain types of cancers.

The term "SCFA" means short chain fatty acid(s). The expression "increased production of SCFA in the colon" means that the amount of SCFA, when measured in the colon (or colon) or part of it, such as the caecum, is higher in an individual who consumed the nutritional composition of the present invention (i.e. containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide), compared to a standard composition (i.e. a nutritional composition not containing at least one N-acetylated oligosaccharide, according to at least one galactooligosaccharide and at least one sialylated oligosaccharide) and/or compared to a standard composition supplemented with conventional fibers like polydextrose or pectin. SCFA may be propionate, butyrate, valerate and/or acetate. In a specific embodiment of the present invention, it is butyrate and/or propionate. The production of SCFA can be measured using techniques known to those skilled in the art, such as gas-liquid chromatography.

The terms "secretion" and "release" can be used interchangeably.

GLP-1 (or GLP1) means glucagon-like peptide 1. It is an incretin secreted by endocrine cells of the intestine known as L-cells. The terms "increased GLP-1 secretion" and "increased GLP-1 release" can be used interchangeably. They mean that the amount of GLP-1 secreted, for example, by the intestinal epithelium, such as in the ileum or colon, among others, is higher in an individual who consumed a nutritional composition according to the present invention (i.e., containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide) compared to a standard composition (i.e., a nutritional composition not containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide). In a specific embodiment, the expression "increased secretion of GLP-1" refers to "increased secretion of GLP-1 in the intestine".

The secretion/release of GLP-1 can be measured using techniques known to those skilled in the art, for example, by measuring its amount in the bloodstream (i.e., by determining the concentration of GLP-1 in blood plasma) in an individual after GLP-1 is secreted into the bloodstream in the absorption of nutrients.

The term "height" refers to the increase in weight, body length and/or head circumference in an infant or young child. In a particular embodiment, it refers to weight. Growth should be understood as the process of change in weight, body length and/or head circumference as an infant or young child matures. In the process of developing an infant, these parameters not only increase. As the standard growth curves published by WHO show, the weight of an infant may decrease in the first days of an infant's life. Thus, growth should be understood as the overall growth of the infant in the first months of life. Therefore, the expressions "growth rate" and "growth rate" can be used as alternatives to the term "growth".

The expressions "promoting normal growth" and "promoting optimal growth" can be used interchangeably. They cover promoting a growth rate that approaches or approximates the growth rate of a breastfed infant. They cover the promotion of growth, which is defined by pediatricians as normal, provided that it is not associated with health problems. These expressions also cover the prevention of overgrowth or overweight, which may occur in formula-fed infants, especially in the first few months of life. The expression "promoting normal growth" can also encompass treatment to control weight and/or avoid weight gain, especially excess weight gain, and/or promote lean mass gain (especially compared to gain in total or adipose tissue mass).

By "mother's milk" is meant the mother's breast milk or colostrum.

The term "oligosaccharide" means a carbohydrate having a degree of polymerization (DP) in the range of 2 to 20 inclusive, but not including lactose. In some embodiments, the carbohydrate has a DP in the range of 3 to 20.

The expressions "at least one N-acetylated oligosaccharide, one galactooligosaccharide and one sialylated oligosaccharide" and "at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide" can be used interchangeably.

The terms "oligosaccharide mixture" or "oligosaccharide mixture" may be used interchangeably. The oligosaccharide mixture according to the present invention contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide. The mixture may be prepared from one or more of these various types of oligosaccharides, i. e. one or more N-acetylated oligosaccharides, one or more galactooligosaccharides, and one or more sialylated oligosaccharides. In some preferred embodiments, the implementation of the oligosaccharides in the mixture of oligosaccharides are cow's milk oligosaccharides (or BMO).

The expression "N-acetylated oligosaccharide" means an oligosaccharide having an N-acetyl residue.

The terms "galacto-oligosaccharide", "galactooligosaccharide", and "GOS" may be used interchangeably. They refer to an oligosaccharide containing two or more galactose molecules that are uncharged and do not contain an N-acetyl residue (i.e. they are neutral oligosaccharides). In a particular embodiment, said two or more galactose molecules are linked by a β-1,2-, β-1,3-, β-1,4- or β-1,6-linkage.

In another embodiment, "galactooligosaccharide" and "GOS" also include oligosaccharides containing one galactose molecule and one glucose molecule (i.e., disaccharides) linked by β-1,2-, β-1,3- or β- 1,6 bond.

The expression "sialylated oligosaccharide" means an oligosaccharide having a sialic acid residue with an associated charge.

The terms "prebiotic", "dietary fiber(s)" and "fiber" may be used interchangeably. They refer to indigestible 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). Microorganism cells are essentially bacteria or yeast.

The term "CFU" should be understood as a colony-forming unit.

Unless otherwise indicated, all percentages given are by weight.

The nutritional composition of the present invention may be present in solid form (eg powder) or in liquid form. The amount of various ingredients (eg, oligosaccharides) may be expressed as g/100 g of the composition on a dry weight basis if the composition is in solid form, such as powder, or as a concentration in g/l of the composition when it is to liquid form (the latter also covers a liquid composition that can be obtained from a powder after reconstitution into a liquid such as milk, water, etc., e.g. grain-based, or any other formula intended for baby food).

Additionally, in the context of the invention, the terms "comprising" or "comprises" do not exclude other possible elements. The composition according to the present invention, including many of the embodiments described herein, may contain the essential elements and limitations of the present invention described herein, as well as any additional or optional ingredients, components or limitations described herein, or otherwise depending on from needs, consist or actually consist of them.

Any reference to prior art documents in this specification should not be construed as an admission that such prior art is widely known or forms part of the common knowledge in the field.

Below is a more detailed description of the invention. It should be noted that the various aspects, features, examples and embodiments described in this application may be compatible and/or combined with each other.

Thus, the first object of the present invention is a nutritional composition containing a mixture of oligosaccharides, and the specified mixture of oligosaccharides contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, for use in reducing and/or avoiding excessive accumulation of adipose tissue mass in an infant or young child by increasing colonic SCFA production and/or by increasing secretion of GLP-1 (eg, GLP-1 secretion in the intestine) in said infant or young child. Said excess accumulation of adipose tissue mass may be an excess accumulation of adipose tissue mass later in life in said infant or young child.

The nutritional composition of the present invention can also be used to prevent any later life health disorder associated with excessive accumulation of adipose tissue mass by increasing colonic SCFA production in an infant or young child. It can be used to prevent obesity or related comorbidities later in life. Some examples of health disorders in later life associated with excessive accumulation of adipose tissue mass are overweight, obesity, insulin resistance, impaired glucose tolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis , hyperuricemia, gallbladder disease, cardiovascular disease, and metabolic syndrome.

As shown in Example 2, the present inventors found that the use of the oligosaccharide mixture of the present invention as a supplement increased the production of SCFAs, especially propionate, butyrate, valerate and acetate, in an animal model. Without wishing to be bound by theory, the inventors of the present invention believe that these particular oligosaccharides act synergistically to unexpectedly provide a substantially increased production of SCFA in the colon. As shown in Example 3, the present inventors also found that this specific mixture of bovine milk oligosaccharides (BMO) increased GLP-1 secretion in an in vitro system using human enteroendocrine cells.

Due to the known properties of SCFA, in particular butyrate and propionate, and GLP-1, for example against obesity, insulin resistance, adipogenesis and food intake, such use of BMO as a supplement may therefore be of interest for use in the prevention of overaccumulation. adipose tissue masses and related health disorders.

The oligosaccharide mixture of the nutritional composition according to the invention contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide. As mentioned above, it may be composed of one or more of these different types of oligosaccharides, ie. one or more N-acetylated oligosaccharides, one or more galactooligosaccharides, and one or more sialylated oligosaccharides. The oligosaccharide mixture of the nutritional composition of the invention may be obtained from the milk of one or more animal species. Milk can be obtained from any mammal, in particular from cows, goats, buffaloes, horses, elephants, camels or sheep.

In an alternative embodiment, the mixture of oligosaccharides can be obtained by purchasing and mixing the individual components.

"N-acetylated oligosaccharide" means an oligosaccharide containing an N-acetylated residue. Suitable N-acetylated oligosaccharides of the nutritional composition oligosaccharide mixture of the present invention include GalNAcβ1.3Galβ1.4Glc and Galβ1.6GalNAcβ1.3Galβ1.4Glc, as well as any mixture thereof. N-acetylated oligosaccharides can be obtained by the action of glucosaminidase and/or galactosaminidase on N-acetylglucose and/or N-acetylgalactose. Equally, N-acetylgalactosyltransferases and/or N-acetylglycosyltransferases can be used for this purpose. N-acetylated oligosaccharides can also be obtained using fermentation technology using appropriate enzymes (recombinant or natural) and/or microbiological fermentation. In the latter case, the microorganisms can either express their natural enzymes and substrates, or microorganisms can be designed to produce the appropriate substrates and enzymes. You can use cultures of microorganisms of the same species or mixed cultures. The formation of N-acetylated oligosaccharides can be initiated by acceptor substrates from any degree of polymerization (DP) from DP=1 onwards. Another option is the chemical conversion of ketohexose (fructose), either free or bound to an oligosaccharide (e.g., lactulose), to N-acetylhexosamine or an oligosaccharide containing N-acetylhexosamine, as described in Wrodnigg, T.M, Dtutz, A.E, Angew . Chem. Int. Ed. 1999:38:827-828.

"Galactooligosaccharide" is an oligosaccharide that contains two or more uncharged galactose molecules and does not contain an N-acetyl residue. Suitable galactooligosaccharides of the oligosaccharide mixture of the nutritional composition according to the present invention include Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.4Glc, Galβ1.3Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.4Glc, Galβ1.3Galβ1.6Galβ1.4Glc, Galβ1.6Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.6Glc, Galβ1.3Galβ1.3Glc, Galβ1.4Galβ1.4Glc and Galβ1.4Galβ1.4Galβ1.4Glc, as well as any mixture thereof. Synthesized galactooligosaccharides such as Galβ1.6Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.6Glc, Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.4Glc, Galβ1.6Galβ1.3Galβ1.4Glc, Galβ1.3Galβ1.6Galβ1.4Glc, Galβ ,4Galβ1,4Glc and Galβ1,4Galβ1,4Galβ1,4Glc and mixtures thereof are commercially available under the tradenames Vivinal® and Elix'or®. Other oligosaccharide suppliers are Dextra Laboratories, Sigma-Aldrich Chemie GmbH and Kyowa Hakko Kogyo Co., Ltd. In an alternative embodiment, specific glycosyltransferases, such as galactosyltransferases, can be used to produce neutral oligosaccharides.

A "sialylated oligosaccharide" is an oligosaccharide containing a sialic acid residue with an associated charge. Suitable sialylated oligosaccharides of the nutritional composition oligosaccharide mixture of the present invention include NeuAcβ2.3Galβ1.4Glc and NeuAcβ2.6Galβ1.4Glc, as well as any mixture thereof. These sialylated oligosaccharides can be isolated from a natural source such as animal milk by chromatography or filtration. In an alternative embodiment of the invention, they can also be produced by biotechnological means using specific sialyltransferases, either through an enzyme-based fermentation technique (recombinant or natural enzymes) or through a microbial fermentation technique. In the latter case, either already existing microorganisms can express their natural enzymes and substrates, or microorganisms can be created that produce the appropriate substrates and enzymes. Single microorganism cultures or mixed cultures can be used. The formation of sialyl oligosaccharides can be initiated by acceptor substrates starting at any degree of polymerization (DP) from DP = 1 onwards.

In one aspect of the invention, the nutritional composition contains a mixture of oligosaccharides in an amount of from 2.5 to 15 wt.%. Alternatively, the nutritional composition contains a mixture of oligosaccharides in an amount of from 3 to 15 wt.%, or in an amount of from 3 to 10 wt.%, or in an amount of from 3.5 to 9.5 wt.%, or in an amount of from 4 up to 9 wt.%, or in an amount of from 4.5 to 8.5 wt.%, or in an amount of from 5.0 to 7.5 wt.%, for example 5 wt.%.

In some specific embodiments, the nutritional composition may contain a mixture of oligosaccharides in an amount of from 0.5 to 3.1 g/100 kcal, or in an amount of from 0.6 to 3.1 g/100 kcal, or in an amount of from 0.6 to 2.0 g/100 kcal, or in an amount of 0.7 to 2.0 g/100 kcal, or in an amount of 0.8 to 1.8 g/100 kcal, or in an amount of 0.9 to 1, 7 g/100 kcal, or in an amount of 1.0 to 1.5 g/100 kcal.

The nutritional composition according to the present invention may contain at least 0.01 wt.% N-acetylated (s) oligosaccharide (s), at least 2.0 wt.% galactooligosaccharide (s) and at least 0.02 wt.% sialylated(s) oligosaccharide(s).

In some embodiments, the implementation of the nutritional composition according to the present invention may contain at least 0.005 wt.%, or at least 0.01 wt.%, or at least 0.02 wt.%, or at least 0.03 wt. %, or at least 0.04 wt.%, or at least 0.05 wt.%, or at least 0.06 wt.% N-acetylated (s) oligosaccharide (s). In some embodiments, it may contain from 0.005 to 0.06 wt.% N-acetylated (s) oligosaccharide (s), for example, from 0.005 to 0.05 wt.%, or from 0.005 to 0.04, or from 0.005 to 0.03 wt.%, or from 0.01 to 0.02 wt.% N-acetylated (s) oligosaccharide (s). A specific example is the amount of 0.01 wt.% N-acetylated (s) oligosaccharide (s).

Additionally, the nutritional composition may contain at least 2 wt.% or at least 3 wt.%, or at least 4 wt.%, or at least 5 wt.%, or at least 5.5 wt.% , or at least 6 wt.%, or at least 7 wt.%, or at least 8 wt.% galactooligosaccharide(s). In some embodiments, it may contain from 4.5 to 8 wt.% galactooligosaccharide(s), for example, from 4.75 to 6 wt.% galactooligosaccharide(s), or from 4.9 to 5 wt.%, or from 5.5 to 6.5 wt.% galactooligosaccharide(s). A specific example is the amount of 4.965 wt.% galactooligosaccharide(s).

Finally, the nutritional composition may contain at least 0.01 wt.%, or at least 0.02 wt.%, or at least 0.03 wt.%, or at least 0.04 wt.%, or at least 0.05 wt.%, or at least 0.06 wt.%, or at least 0.07 wt.%, or at least 0.08 wt.%, or at least 0.09 wt.% sialylated oligosaccharides. In some embodiments, it may contain from 0.02 to 0.09 wt.% sialylated(s) oligosaccharide(s), for example, from 0.02 to 0.07 wt.% sialylated(s) oligosaccharide(s). ), or from 0.02 to 0.05 wt.% sialylated(s) oligosaccharide(s), or from 0.003 to 0.07 wt.% sialylated(s) oligosaccharide(s). A specific example is the amount of 0.025 wt.% sialylated(s) oligosaccharide(s).

In a specific embodiment, the nutritional composition according to the present invention may contain from 0.01 to 0.07 wt.% N-acetylated (s) oligosaccharide (s), from 2.0 to 8.0 wt.% galactooligosaccharide (s ) and from 0.02 to 0.09 wt.% sialylated(s) oligosaccharide(s).

In another specific embodiment, the nutritional composition in accordance with the present invention may include from 0.01 to 0.03 wt.% N-acetylated (s) oligosaccharide (s), 5.95 wt.% galactooligosaccharide (s) and from 0.02 to 0.09 wt.% sialylated(s) oligosaccharide(s).

In a specific embodiment, the nutritional composition may include from 0.0015 to 0.005 g/100 kcal of N-acetylated oligosaccharide(s), from 0.70 to 1.5 g/100 kcal of galactooligosaccharide(s), and from 0.0045 to 0.0085 g/100 kcal of sialylated oligosaccharide(s).

In another specific embodiment, the nutritional composition may include from 0.0015 to 0.0045 g/100 kcal of N-acetylated oligosaccharide(s), from 0.74 to 1.2 g/100 kcal of galactooligosaccharide(s). ) and from 0.0045 to 0.0075 g/100 kcal of the sialylated oligosaccharide(s).

In a particularly preferred embodiment, the mixture of oligosaccharides in the nutritional composition according to the present invention contains from 0.1 to 4.0 wt.% N-acetylated (s) oligosaccharide (s), from 92.0 to 99.5 wt.% galactooligosaccharide (s) and from 0.2 to 4.0 wt.% sialylated (s) oligosaccharide (s).

The nutritional composition according to the invention may also contain other types of prebiotics (ie other than oligosaccharides and in addition to the oligosaccharides contained in the mixture of oligosaccharides defined in accordance with the present invention). Examples of other types of prebiotics include human milk oligosaccharides (HMOs), such as fucosylated oligosaccharides; oligofructose; fructooligosaccharides (FOS); inulin xylooligosaccharides (XOS); polydextrose or any mixture thereof.

"Fucosylated oligosaccharide" is a human milk oligosaccharide having a fucose residue. It has a neutral character. Some examples are 2'-FL (2'-fucosyllactose or 2-fucosyllactose or 2FL or 2-FL), 3-FL (3-fucosyllactose), difucosyllactose, lacto-N-fucopentaose (e.g. lacto-N- fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V), lacto-N-fucohexaose, lacto-N-difucohexaose I, fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose, difucosyl lacto-N-hexaose I, difucosyl lacto-N-neohexaose II, and any combination thereof.

Suitable commercially available products that can be used in addition to the oligosaccharides contained in the mixture of oligosaccharides to obtain nutritional compositions according to the present invention include combinations of FOS with inulin, such as the product sold by BENEO under the trade name Orafti, or polydextrose, marketed by Tate & Lyle under the tradename STA-LITE®.

The nutritional composition of the present invention may further comprise at least one probiotic (or probiotic strain), such as a probiotic bacterial strain.

The most commonly used probiotic microorganisms are predominantly bacteria and yeasts of the following genera: Lactobacillus spp., Streptococcus spp., Enterococcus spp., Bifidobacterium spp. and Saccharomyces spp.

In some specific embodiments, the implementation of the probiotic is a probiotic bacterial strain. In some specific embodiments, the implementation is, in particular, strains of Bifidobacteria and/or Lactobacilli.

Подходящие пробиотические штаммы включают в себя 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, Bifidobacterium adolescentis or any mixture thereof.

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 & Gamble Co. under the Bifantis trademark; and Bifidobacterium breve sold by Institut Rosell (Lallemand) under the trademark R0070.

The nutritional composition according to the invention may contain from 10e3 to 10e12 CFU of the probiotic strain, more preferably from 10e7 to 10e12 CFU, for example from 10e8 to 10e10 CFU of the probiotic strain per gram of composition, based on dry weight.

In one embodiment, the probiotics are viable. In another embodiment of the present invention, the probiotics are non-propagating or inactivated. In some other embodiments, both viable probiotics and inactivated probiotics may be used.

The nutritional composition of the invention may further comprise at least one phage (bacteriophage) or a mixture of phages, preferably directed against pathogenic microorganisms Streptococci, Haemophilus, Moraxella and Staphylococci.

A nutritional composition according to the invention may be, for example, an infant formula, initial infant formula, complementary formula, infant formula, a cereal-based infant formula, a fortifier such as a breast milk fortifier, or a supplement. In some specific embodiments, the composition of the invention is an infant formula, fortifier or supplement, which may be for the first 4 or 6 months of life. In a preferred embodiment, the nutritional composition of the invention is an infant formula.

In some other embodiments, the nutritional composition of the present invention is a fortifier. The fortifier can be a human milk fortifier (eg, human breast milk fortifier) or a formula fortifier, such as an infant formula fortifier or formula fortifier for formula fed/follow-up formula.

If the nutritional composition is a supplement, it may be provided in unit dosage form.

The nutritional composition of the present invention may be in solid (eg powder), liquid or jelly form.

The nutritional compositions of the present invention, and especially infant formulas, typically contain a protein source, a carbohydrate source, and a lipid source.

The nutritional composition of the present invention typically contains a protein source. The protein may be present in an amount of 1.5 to 3 g per 100 kcal. In some embodiments, particularly when the composition is for preterm infants, the amount may be from 2.4 to 4 g/100 kcal, or more than 3.6 g/100 kcal. In some other embodiments, the amount of protein may be less than 2.0 g per 100 kcal, such as 1.8 to 2 g/100 kcal, or less than 1.8 g per 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 ratios.

In some preferred embodiments, the protein source is dominated by whey (i.e., more than 50% of the proteins are derived from whey proteins, such as 60% or 70%).

The proteins may be native or hydrolysed, or they may be a mixture of native and hydrolyzed proteins. The term "native" means that the bulk of the proteins are native, i.e. their molecular structure is not altered, e.g. at least 80% of the proteins are not altered, e.g. at least 85% of the proteins are not altered, preferably at least 90% of the proteins are not altered, even more preferably at least 95% of the proteins are not altered, e.g. at least 98% of the proteins are changed. In a specific embodiment, 100% of the proteins are unchanged.

The term "hydrolyzed" in the context of the present invention means a protein that has been hydrolyzed or cleaved into its constituent amino acids.

Proteins may be fully or partially hydrolysed. It may be desirable to provide partially hydrolyzed proteins (degree of hydrolysis between 2 and 20%), for example for infants or young children who are at risk of developing cow's milk allergy. If it is necessary to use hydrolyzed proteins, the hydrolysis process can be carried out as desired and in a manner known in the art. For example, whey protein hydrolysates can be obtained by enzymatic hydrolysis of a whey fraction in one or more steps. If the whey fraction used as starting material is substantially free of lactose, it has been found that the blocking of protein lysine occurs to a much lesser extent during hydrolysis. This reduces the degree of blocking of lysine from about 15 wt.% total lysine to less than about 10 wt.% lysine; for example, about 7 wt.% lysine, which significantly improves the nutritional quality of the protein source.

In one embodiment of the invention, at least 70% of the proteins are hydrolyzed, preferably at least 80% of the proteins are hydrolyzed, e.g. at least 85% of the proteins are hydrolyzed, even more preferably at least 90% of the proteins are hydrolyzed, e.g. at least 95% of the proteins are hydrolyzed in particular, at least 98% of the proteins are hydrolyzed. In a specific embodiment, 100% of the proteins are hydrolysed.

In one specific embodiment, the proteins of the nutritional composition are hydrolyzed, fully hydrolyzed, or partially hydrolyzed. The degree of hydrolysis (DH) of the protein can be from 8 to 40, or from 20 to 60, or from 20 to 80, or more than 10, 20, 40, 60, 80, or 90.

In a specific embodiment, the nutritional composition according to the invention is a hypoallergenic composition. In another specific embodiment, the composition according to the invention is a hypoallergenic nutritional composition.

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

The nutritional composition according to the present invention typically 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 may be any lipid or fat that is suitable for use in infant formulas. Some suitable fat sources include palm oil, high oleic sunflower oil, and high oleic safflower oil. You can also add essential fatty acids - linoleic 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. The ratio of n-6 to n-3 fatty acids in the fat source can be from about 5:1 to about 15:1; for example, from about 8:1 to about 10:1.

The nutritional composition of the invention may also contain all the vitamins and minerals that are considered indispensable in the daily diet, in nutritionally significant amounts. Some vitamins and minerals have minimum requirements. Examples of minerals, vitamins and other nutrients that are optionally present in the composition of the present invention include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folate 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 nutritional composition of the present invention may contain emulsifiers and stabilizers such as soy, lecithin, mono- and diglyceryl esters of citric acid, and the like.

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

The nutritional composition according to the present invention may also contain carotenoid(s).

The nutritional composition of the invention (eg, infant formula) can be obtained by mixing a protein source, a carbohydrate source, and a fat source in appropriate proportions. Emulsifiers can be added if desired. Vitamins and minerals can be added at this stage, but are usually added later to prevent thermal decomposition. Any freeze-dried vitamins, emulsifiers, and the like may be dissolved in the fat source prior to mixing. Water, preferably reverse osmosis water, can then be mixed in to form a liquid mixture.

The liquid mixture can then be heat treated to reduce bacterial loads. For example, the liquid mixture can be rapidly heated to a temperature in the range from about 80°C to about 110°C for about 5 seconds to about 5 minutes. This can be done by steam injection or by means of a heat exchanger such as a plate heat exchanger.

The liquid mixture can then be cooled to a temperature of from about 60° C. to about 85° C., for example, by flash cooling. The liquid mixture can then be homogenized, for example, in two stages: under pressure from about 7 MPa to about 40 MPa in the first stage and from about 2 MPa to about 14 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, the pH and solids content of the homogenized mixture can be standardized for convenience.

The homogenized mixture can be 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% wt.

The mixture of oligosaccharides can be obtained by any suitable method known in this field, and added at various stages in the process of obtaining a nutritional composition of the present invention. The oligosaccharide mixture can be added directly to the nutritional composition (eg, infant formula) by dry mixing (ie, during the mixing step). In an alternative embodiment, the mixture of oligosaccharides can be added to the liquid mixture before heat treatment to reduce the bacterial load. The individual components of the oligosaccharide mixture can also be added separately to the nutritional composition, in which case the oligosaccharide mixture is preferably added to the liquid phase just prior to drying.

The nutritional composition according to the present invention is intended for use in infants or young children. Infants or young children may be born at term or prematurely. In a specific embodiment, the nutritional composition of the invention is for use in infants or young children who were born prematurely. In a specific embodiment, the nutritional composition of the invention is for use in preterm infants.

The nutritional composition of the present invention may also be administered to an infant or young child who has been delivered by caesarean section or by vaginal delivery.

In some embodiments, the implementation of the nutritional composition according to the present invention may be intended for use before and/or during the period of weaning.

In some embodiments, the nutritional composition of the present invention is for use in infants or young children who are at risk of developing a health disorder later in life associated with excessive accumulation of adipose tissue mass.

Premature infants may be at increased risk of developing insulin resistance, hyperglycemia, malnutrition, impaired growth in lean body mass, fat accumulation in the viscera, and metabolic disorders later in life.

It may also target infants or young children who are at risk of becoming overweight or obese later in life.

In some embodiments, the nutritional composition of the present invention is for use in infants or young children born to overweight and obese women. Indeed, scientific evidence consistently confirms that babies born to overweight and obese mothers have a higher risk of becoming overweight or obese later in life than babies born to mothers who are not overweight or obese. In some embodiments, the nutritional composition of the present invention is for use in infants or young children born to mothers who have experienced gestational diabetes.

In a specific example, the nutritional composition according to the present invention can be used in infants or young children who have had IUGR (intrauterine growth retardation). This particular population is at risk of developing a health disorder later in life associated with excessive accumulation of adipose tissue mass, since they will have a higher appetite to compensate for their growth retardation. But they may not be healthy fed using standard recipes, for example, they may have a higher total mass and/or increase in adipose tissue mass compared to an increase in lean mass, which may lead to development and programming for future health conditions, including obesity in later life or future associated comorbidities. The nutritional composition of the present invention is believed to provide normal growth.

The age for administration (or use or feeding) of a nutritional composition, as well as the duration of such administration, depends on the possibilities and needs.

Since the nutritional composition is used in particular for the purpose of prophylaxis (prevention of a health disorder later in life), it can, for example, be given immediately after the birth of babies. The composition according to the present invention can also be given during the first week of an infant's life, or during the first 2 weeks of life, or during the first 3 weeks of life, or during the first month of life, or during the first 2 months of life, or during the first 3 months of life, or during the first 4 months of life, or during the first 6 months of life, or during the first 8 months of life, or during the first 10 months of life, or during the first year of life, or during the first two years of life, or even longer. In some particularly advantageous embodiments of the present invention, the nutritional composition is given (or administered) to an infant during the first 4 or 6 months after the birth of said infant.

In some other embodiments, the nutritional composition of the present invention is given several days (e.g., 1, 2, 3, 5, 10, 15, 20...) or several weeks (e.g., 1, 2, 3, 4, 5, 6 ,7, 8, 9, 10...), or a few months (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10...) after birth. First of all, this may be relevant, but not necessarily, in the case when the baby was born prematurely.

In one embodiment, the composition of the invention is given to an infant or young child as a composition in addition to mother's milk. In some embodiments, the infant or young child receives breast milk for at least the first 2 weeks, the first 1, 2, 4, or 6 months. In one embodiment, the nutritional composition of the invention is given to an infant or young child after such a breast milk feeding period, or is given during such a breast milk feeding period. In another embodiment, the composition is given to an infant or young child as the sole or primary nutritional composition for at least one period of time, such as after the 1st, 2nd, or 4th month of life, for at least 1, 2, 4 or 6 months.

In one embodiment, the nutritional composition of the invention is a complete nutritional composition (satisfying all or most of the nutritional needs of the subject). In another embodiment, the nutritional composition is a supplement or fortifier intended, for example, to supplement breast milk or supplement an infant formula or formula for feeding babies.

The mixture of oligosaccharides present in the nutritional composition of the invention may be obtained from the milk of one or more animal species. Milk can be obtained from any mammal, in particular from cows, goats, buffaloes, horses, elephants, camels or sheep. In a specific embodiment, the oligosaccharides of the oligosaccharide mixture are bovid milk oligosaccharides and can be obtained from cow, goat or buffalo milk. In a preferred embodiment, the oligosaccharides are derived from cow's milk. Documents WO 2006087391 and WO 2012160080 give several examples of the preparation of a BMOS mixture.

The present inventors found that BMO intervention in an animal model increased their colonic SCFA production, especially caecal SCFA production, and in particular butyrate and propionate. Since these SCFAs have been shown to protect against obesity, insulin resistance, regulate adipogenesis and food intake, the nutritional composition of the present invention would therefore be useful in reducing and/or avoiding excessive accumulation of adipose tissue mass in an infant or young child by increasing colonic SCFA production in said infant or young child.

It can be used to prevent a later life disorder associated with excess fat accumulation in an infant or young child by increasing colonic SCFA production in said infant or young child.

The production of one or more SCFAs may be increased. SCFA may be propionate, butyrate, valerate and/or acetate. In a particular embodiment, the SCFA is propionate and/or butyrate (ie, propionate, butyrate, or both).

Prevention of excess fat accumulation and associated health problems later in life can be done in a variety of ways. Increasing the production of SCFA represents a new clinical situation in which the goal of prevention can be achieved in a new way.

Therefore, in a particular embodiment, prevention of excess fat accumulation and associated health disorders later in life is achieved by increasing colonic production of propionate and/or butyrate in said infant or young child.

In a specific embodiment, the production of propionate and/or butyrate is measured using gas-liquid chromatography, and it can be expressed in nmol/mg dry weight.

In a specific embodiment, colonic butyrate production is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% of the butyrate production in the colon obtained with nutritional composition without at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide.

In a specific embodiment, colonic butyrate production is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 100% or more compared to with the production of butyrate in the colon, obtained with a nutritional composition supplemented with conventional fibers like polydextrose or pectin.

In a particular embodiment, colonic propionate production is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, compared to colonic propionate production obtained with a nutritional composition without at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide .

In a particular embodiment, colonic propionate production is increased by at least 10%, or at least 15%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70% compared to colonic propionate production obtained with a nutritional composition supplemented with conventional fibers like polydextrose or pectin.

The present inventors also found that the BMO mixture significantly increased GLP-1 secretion in an in vitro system using NCl-H716 cells. Since GLP-1 has been specifically shown to improve glucose clearance, slow gastric emptying, reduce appetite and food intake, reduce body weight, and provide some beneficial effects on the cardiovascular system, the nutritional composition of the present invention will therefore be useful in reducing and/or avoiding excessive accumulation of adipose tissue mass in an infant or young child. It can be used in an infant or young child to prevent a health disorder later in life like later life obesity, diabetes... by increasing GLP-1 secretion (especially GLP-1 secretion in the gut) in said infant or young child.

In a particular embodiment, GLP-1 secretion is increased by the BMO mixture of the present invention by at least 10%, or at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% compared to the secretion of GLP-1 obtained using a nutritional composition without said mixture BMO.

The health disorder later in life that may be prevented by the nutritional composition of the present invention may be any type of health condition associated with excessive accumulation of adipose tissue mass and which may be affected, directly or indirectly, by the amount of SCFA in the colon. and/or GLP-1 secretion.

As mentioned in the Background of the Invention section, SCFAs (especially butyrate and propionate) and GLP-1 are known to protect against obesity, insulin resistance, regulate adipogenesis and food intake. Later life health disorders associated with excessive accumulation of adipose tissue mass, the prevention of which can be carried out using a nutritional composition according to the present invention, can be selected from a list consisting of overweight, obesity or obesity-related comorbidities. Some examples of comorbidities associated with obesity later in life are: insulin resistance, impaired glucose tolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gallbladder disease, cardiovascular vascular disease, metabolic syndrome, and certain types of cancer.

Thus, in some embodiments, a nutritional composition of the present invention may be used to prevent a health disorder later in life selected from a list consisting of overweight, obesity, type 2 diabetes, insulin resistance, hypertension, cardiovascular disease, or metabolic syndrome. . In a preferred embodiment, it is used to prevent obesity later in life.

Another object of the present invention relates to the use of a nutritional composition according to the present invention to reduce the risk of developing overweight later in life in an infant or young child.

Another object of the present invention relates to a nutritional composition according to the present invention for use in promoting normal growth in an infant or young child by increasing the production of SCFA in the colon (especially butyrate and propionate) and/or by increasing the secretion of GLP-1 in said infant or a young child, in particular to avoid excess weight gain in said infant or young child.

Other objects

Another object of the present invention is the use of a mixture of oligosaccharides containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, in the preparation of a nutritional composition for reducing and/or avoiding excessive accumulation of adipose tissue mass in an infant or of a young child and/or to prevent a health disorder later in life associated with excessive accumulation of adipose tissue mass in an infant or young child by increasing the production of SCFA in the colon of said infant or young child, especially butyrate and/or propionate, and/or by increasing GLP-1 secretion in said infant or young child.

Another object of the present invention is the use of a mixture of oligosaccharides containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, in the preparation of a nutritional composition for promoting normal growth in an infant or young child by increasing the production of SCFA in the colon of said infant or young child, especially butyrate and/or propionate, and/or by increasing secretion of GLP-1 in said infant or young child.

Another object of the present invention is the use of a mixture of oligosaccharides containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, (or the use of a nutritional composition containing such a mixture of oligosaccharides) to increase the production of SCFA in the colon in an infant or young child, especially butyrate and/or propionate, and/or to increase GLP-1 secretion in an infant or young child.

Another object of the present invention is the use of a nutritional composition containing a mixture of oligosaccharides, which contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, for the prevention of overweight in an infant or young child by increasing the production SCFA in the colon of said infant or young child, especially butyrate and/or propionate, and/or by increasing secretion of GLP-1 in said infant or young child.

Another object of the present invention is the use of an oligosaccharide mixture containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, to promote normal growth in an infant or young child by increasing the production of SCFA in the colon in said infant or young child, especially butyrate and/or propionate, and/or by increasing secretion of GLP-1 in said infant or young child.

Another object of the present invention is a pharmaceutical composition containing a mixture of oligosaccharides, which contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, for use in reducing and/or avoiding excessive accumulation of adipose tissue mass in infant or young child and/or for use in the prevention of a health disorder later in life associated with excess fat accumulation in an infant or young child, by increasing the production of SCFA in the colon of said infant or young child, especially butyrate and /or propionate, and/or by increasing the secretion of GLP-1 in said infant or young child.

Another object of the present invention is a method for reducing and/or avoiding excessive accumulation of adipose tissue mass in an infant or young child and/or for the prevention of health disorders in later life associated with the accumulation of fat in an infant or young child, by increasing the production of SCFA in the colon. gut of said infant or young child, especially butyrate and/or propionate, and/or by increasing secretion of GLP-1 in said infant or young child, said method comprising administering to said infant or young child a nutritional composition comprising a mixture of oligosaccharides that contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide.

Another object of the present invention is a method of promoting normal growth in an infant or young child by increasing the production of SCFA in the colon of said infant or young child, in particular butyrate and/or propionate, and/or by increasing the secretion of GLP-1 in said an infant or young child, said method comprising administering to said infant or young child a nutritional composition containing an oligosaccharide mixture that contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide.

Another object of the present invention is a method for increasing the production of SCFA in the colon in an infant or young child, in particular butyrate and/or propionate, and/or for increasing the secretion of GLP-1 in an infant or young child, which method comprises administering to said infant or young child a nutritional composition containing an oligosaccharide mixture that contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide.

Another object of the present invention is an oligosaccharide mixture containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, for use in a nutritional composition for an infant or young child as a therapeutic agent that increases the production SCFA in the colon of said infant or young child, especially butyrate and/or propionate, and/or which increases GLP-1 secretion in said infant or young child.

Another object of the present invention is a nutritional composition containing at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, for use as a therapeutic agent that increases the production of SCFA in the colon in an infant or young child. age, especially butyrate and/or propionate, and/or which increases GLP-1 secretion in an infant or young child.

The previously mentioned embodiments and examples (eg, relating to types and amounts of oligosaccharide, nutritional composition, administration, target population...) also apply to all other subjects.

Examples

The following examples demonstrate some specific embodiments of the composition for use in accordance with the present invention. These examples are provided for illustrative purposes only and should not be construed as limiting the present invention, as many variations are possible within the spirit of the invention.

Example 1

Table 1 below shows an example of the composition of an infant formula containing a mixture of oligosaccharides according to the present invention. The mixture of oligosaccharides may contain, for example, from 0.1 to 4.0 wt.% N-acetylated (s) oligosaccharide (s), from 92.0 to 99.5 wt.% galactooligosaccharide (s) and from 0 .2 to 4.0 wt.% sialylated(s) oligosaccharide(s).

Table 1. An example of the composition of the nutritional composition (for example, infant formula) in accordance with the present invention

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 fifty Mn (mcg) eight fifty Se (mcg) 2 13 Vitamin A (µg RE) 105 700 Vitamin D (mcg) 1.5 ten Vitamin E (mg ET) 0.8 5.4 Vitamin K1 (mcg) eight 54 Vitamin C (mg) ten 67 Vitamin B1 (mg) 0.07 0.47 Vitamin B2 (mg) 0.15 1.0 Niacin (mg) one 6.7 Vitamin B6 (mg) 0.075 0.50 Folic acid (mcg) 9 60 Pantothenic acid (mg) 0.45 3 Vitamin B12 (mcg) 0.3 2 Biotin (mcg) 2.2 fifteen Choline (mg) ten 67 Fe (mg) 1.2 eight I (µg) fifteen 100 Cu (mg) 0.06 0.4 Zn (mg) 0.75 5 Oligosaccharide Blend (g) 1.38 9.0

Example 2

Study Description

Female BALB/cByJ CRL mice from Charles River at 5 weeks of age were divided into several groups and fed for 6 weeks based on the following protocol:

- Week 1: low fiber diet (composition detailed in Table 2) for all groups

- Weeks 2-6:

• Control group (group A): low fiber diet (same as week 1)

• Test groups (groups B-D): low fiber diet (same as week 1) supplemented with 5% by weight of test fiber (5% of total low fiber diet was replaced with 5% of test fiber)

Table 2. Composition of a low-fiber diet

Essential Nutrients Dry matter 93.9% Crude protein 18.0% Crude fat 5.0% Unrefined fiber 0.3% Crude minerals 3.5% Nitrogen Free Extract (NFE) 67.1% Gross energy 17.7 MJ/kg Metabolic energy 16.1 MJ/kg Starch 42.5% Amino acids Arginine 0.76% Lysine 1.66% Methionine 0.60% Methionine + cysteine 0.97% Tryptophan 0.28% Threonine 0.92% Macronutrients Calcium 0.62% Phosphorus 0.33% Magnesium 0.06% Sodium 0.24% Potassium 0.41% Chlorine 0.58% trace elements Iron 50 mg/kg Zinc 37 mg/kg Copper 6 mg/kg Iodine 0.6 mg/kg Manganese 12 mg/kg Selenium 0.22 mg/kg Added vitamins Vitamin A 4000 IE|UI|IU/kg Vitamin D3 1000 IE|UI|IU/kg Vitamin E 100 mg/kg Vitamin K3 4 mg/kg Vitamin B1 5 mg/kg Vitamin B2 6 mg/kg Vitamin B6 6 mg/kg Vitamin B12 0.05 mg/kg Nicotinic acid 32 mg/kg Pantothenic acid 16 mg/kg Folic acid 2 mg/kg Biotin 0.2 mg/kg Choline 998 mg/kg

The following fibers were studied:

BMO = cow's milk oligosaccharides: a mixture of oligosaccharides containing 99.3% GOS, 0.2% N-acetylated oligosaccharide and 0.5% sialylated oligosaccharide was studied

PDX = polydextrose

Pectin

Table 3 provides a summary of the various study groups and diets.

Table 3. Test groups and diets in the study

Group group label Diet Sample size A Positive counter. or counter. positive Diet low in fiber eight B BMOS Low fiber diet + 5 wt% BMO eight C PDX Low fiber diet + 5 wt% polydextrose eight E Pectin Low fiber diet + 5 wt% pectin eight

After 6 weeks, the animals in each group were sacrificed and the contents from the caecum were collected. SCFA production was measured by gas liquid chromatography (GLC; SCFA amounts in nmol/mg dry weight). The following SCFAs were measured: propionate, butyrate, valerate and acetate.

The measurement was carried out based on the following protocol: SCFA in acidic solution (pH 2.0-3.0) was separated on a GLC column coated with a polar stationary phase. This allowed for minimal sample preparation (no derivatization) and direct basic detection using a FID (flame ionization detector). SCFA was extracted from the caecum using an acid phosphate buffer containing HgCl2 to inactivate any residual bacterial activity and an internal standard (2,2-dimethyl-butyric acid) for GLC analysis. After centrifugation, the filter-sterilized supernatant was ready for GLC analysis. The SCFA content was measured simultaneously.

Ratios of median values were calculated to compare different fiber-fortified diets in terms of SCFA production.

results

Butyrate production was significantly increased by the BMO-enriched diet (see FIG. 1). Its production increased by approximately 99% compared to the positive control. Its production increased by 133% and 136% compared to pectin and PDX, respectively.

Propionate production was significantly increased by the BMO-enriched diet (see FIG. 2). Its production increased by approximately 69% compared to the positive control. Its production increased by 74% and 75% compared to pectin and PDX, respectively.

These results are very surprising since pectin has generally been regarded as a strong SCFA inducer (Stark et al, J Nutr. 1993, In vitro production of short-chain fatty acids by bacterial fermentation of dietary fiber compared with effects of those fibers on hepatic sterol synthesis in rats; Yang et al, Anaerobe, 2013, In vitro characterization of the impact of selected dietary fibers on fecal microbiota composition and short chain fatty acid production).

In FIG. 3 shows the ratio of the median content of each test SCFA in each fiber-enriched diet divided by the median value for the positive control diet (i.e., low fiber diet only). A ratio of 1 (black line) means that there is no difference between the fortified diet and the control diet. A ratio below 1 means that the content of the corresponding SCFA is higher in the control diet compared to the fiber-enriched diet, while a ratio above 1 means that the content of the corresponding SCFA is higher in the fiber-fortified diet than in the control. Diets enriched with PDX and pectin induced less release of SCFA of all species. On the other hand, a BMOS-enriched diet induces greater release of all types of SCFA (acetate, propionate, butyrate, valerate) than a low-fiber diet and other fibers studied. The BMOS-enriched diet was the only one that stimulated butyrate and propionate production so strongly.

Thus, the present inventors unexpectedly found that mice fed a specific BMO mixture had higher production of all SCFAs studied in the caecum (and therefore in the colon), and especially in the case of butyrate and propionate.

Since SCFAs, and in particular butyrate and propionate, are known to protect against obesity, insulin resistance, regulate adipogenesis, food intake, and are involved in the prevention of non-alcoholic fatty liver disease or conditions associated with cardiac metabolism, such as the development of atherosclerosis and inflammation, the composition containing a mixture of oligosaccharides that contains at least one N-acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, therefore, will be effective in infants or young children for use in avoiding excessive accumulation of adipose tissue mass and in prevention of health disorder in later life associated with it, by increasing the production of SCFA in the colon in these infants or young children. This represents a new method of targeted prevention of these specific health conditions.

Example 3

Study Description

Cell line and culture conditions

Human NCI-H716 cells were obtained from the American Type Culture Collection. These cells are human enteroendocrine cells capable of secreting GLP-1.

To maintain proliferation, cells were grown in suspension in Gibco® RPMI 1640 medium supplemented with 10% FBS (fetal bovine serum), 2 mM l-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin. Adhesion and endocrine differentiation were initiated by growing cells in Matrigel coated dishes in DMEM (Dulbecco's Modified Eagle's Medium) with high glucose, 10% FBS, 2 mM l-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin.

Secretion studies

Two days prior to experiments, 1x10 ⁶ NCI-H716 cells were seeded in 12-well culture plates coated with Matrigel gel. On the day of the experiment, the supernatants were replaced with KRB buffer (Krebs-Ringer bicarbonate buffer) with or without the test agent:

KRB buffer only = negative control

KRB buffer + 1 µM PMA (phorbol-12-myristate-13-acetate) = positive control

KRB buffer + 10 mg/ml BMO = experimental sample. BMO formulation: oligosaccharide mixture containing 99.3% GOS, 0.2% N-acetylated oligosaccharide and 0.5% sialylated oligosaccharide.

Cells were incubated for 2 hours at 37°C with various effectors. 2-hour incubation of cells with different effectors did not affect cell viability. Supernatants were collected with the addition of a complete cocktail of EDTA free protease inhibitors, DPP-IV inhibitor (dipeptidyl peptidase 4) and frozen at -80°C for further analysis. Cells were scraped and sonicated in a homogenization assay. Cells were scraped and sonicated for normalization. To normalize the content of GLP-1, proteins were measured in the cell homogenate. Biologically active GLP-1 was measured by an enzyme-linked immunosorbent assay (ELISA) as described by the distributor of the GLP-1 (active) ELISA assay kit (EMD Millipore; # EGLP-35K).

results

GLP-1 secretion was significantly increased when BMOS was added to this in vitro system using human NCl-H716 cells (P<0.01 vs. negative control; two-tailed t-test). Its secretion increased, in particular, by 92% compared to the negative control.

Since GLP-1 is known to improve glucose clearance, slow gastric emptying, reduce appetite and food intake, reduce body weight, and provide some beneficial effects on the cardiovascular system, a composition containing an oligosaccharide mixture that contains at least one N -acetylated oligosaccharide, at least one galactooligosaccharide and at least one sialylated oligosaccharide, therefore, will be effective in infants or young children for use in avoiding excessive accumulation of adipose tissue mass and in preventing later life health disorders like obesity and diabetes, by increasing the secretion of GLP-1 in said infants or young children. This represents a new method of targeted prevention of these specific health conditions.

Claims (14)

1. The use of an artificial nutritional composition containing a mixture of oligosaccharides to reduce and/or avoid excessive accumulation of adipose tissue in an infant or young child and/or to prevent health disorders in later life associated with excessive accumulation of adipose tissue in an infant or child younger age selected from a list consisting of insulin resistance, impaired glucose tolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gallbladder disease, cardiovascular disease, and metabolic syndrome , by increasing colonic SCFA production and/or by increasing GLP-1 secretion in said infant or young child, wherein said oligosaccharide mixture comprises at least one N-acetylated oligosaccharide, at least one galactooligosaccharide, and at least one sialylated oligosaccharide. 2. Use according to claim 1, wherein the N-acetylated oligosaccharide is GalNAcβ1.3Galβ1.4Glc, Galβ1.6GalNAcβ1.3Galβ1.4Glc or a mixture thereof. 3. Use according to any of the preceding claims, wherein the galactooligosaccharide is Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.4Glc, Galβ1.3Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.4Glc, Galβ1.3Galβ1.6Galβ1.4Glc, Galβ1 ,6Galβ1.3Galβ1.4Glc, Galβ1.6Galβ1.6Galβ1.6Glc, Galβ1.3Galβ1.3Glc, Galβ1.4Galβ1.4Glc, Galβ1.4Galβ1.4Galβ1.4Glc or a mixture thereof. 4. Use according to any one of the preceding claims, wherein the sialylated oligosaccharide is NeuAcβ2.3Galβ1.4Glc, NeuAcβ2.6Galβ1.4Glc or a mixture thereof. 5. Use according to any one of the preceding claims, wherein the artificial nutritional composition contains a mixture of oligosaccharides in an amount of 2.5 to 15.0% by weight. 6. Use according to any of the preceding claims, wherein the artificial nutritional composition contains at least 0.01 wt.% N-acetylated (s) oligosaccharide (s), at least 2.0 wt.% galactooligosaccharide (s) and at least 0.02 wt.% sialylated(s) oligosaccharide(s). 7. Use according to any one of the preceding claims, wherein the mixture of oligosaccharides contains from 0.1 to 4.0 wt.% N-acetylated (s) oligosaccharide (s), from 92.0 to 99.5 wt.% galactooligosaccharide ( -s) and from 0.2 to 4.0 wt.% sialylated (s) oligosaccharide (s). 8. Use according to any one of the preceding claims, wherein the artificial nutritional composition is an infant formula. 9. Use according to any one of the preceding claims, wherein the artificial nutritional composition is administered to an infant during the first six months of life. 10. Use according to any one of the preceding claims, characterized in that the mixture of oligosaccharides is derived from animal milk. 11. Use according to any one of the preceding claims, wherein the SCFA is selected from the list consisting of propionate, butyrate, valerate, acetate, or a combination thereof, preferably butyrate and/or propionate. 12. Use according to any one of the preceding claims, wherein the infant or young child is at risk of developing a health disorder later in life associated with excessive accumulation of adipose tissue mass, wherein the infant or young child is at particular risk of developing obesity later in life. 13. Use according to any one of the preceding claims to promote normal growth in an infant or young child by increasing colonic SCFA production and/or by increasing GLP-1 secretion in said infant or young child. 14. Use according to claim 13 to avoid excess weight gain in said infant or young child.

Images