Patent Application
20190046545
This invention relates to nutritional compositions such as infant formula, comprising an oligosaccharide mixture which is specifically designed to avoid excessive fat mass accumulation and to prevent related later in life health disorders in infants or young children like obesity later in life, by increasing colonic SOFA production and/or GLP-1 secretion in said infants or young children.
Scientific evidence suggests that infancy may be a critical period in the development and programming for future health conditions including later in life obesity or future related comorbidities (including the metabolic disorders).
Overweight and obesity are defined as abnormal or excessive fat accumulation that may impair health. Body mass index (BMI) is a simple index of weight-for-height that is commonly used to classify overweight and obesity in adults. It is defined as a person's weight in kilograms divided by the square of his height in meters (kg/m2). The WHO definition is: a BMI greater than or equal to 25 is overweight; a BMI greater than or equal to 30 is obesity. The prevalence of obesity and overweight in adults, children and adolescents has increased rapidly over the past 30 years globally and continues to rise. Worldwide obesity has more than doubled since 1980, as reported by the WHO. It has become a global health concern since it is associated with a reduced life time, an altered life quality and it is responsible of further health conditions. Overweight and obesity are linked to more deaths worldwide than underweight. Childhood obesity is indeed associated with a higher chance 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 psychological effects. Raised BMI is a major risk factor for noncommunicable diseases such as cardiovascular diseases (mainly heart disease and stroke), which were the leading cause of death in 2012; diabetes; musculoskeletal disorders (especially osteoarthritis—a highly disabling degenerative disease of the joints); even some cancers (endometrial, breast, and colon).
Insulin resistance can lead to type 2 diabetes. Type 2 diabetes is a chronic disease associated with abnormally high levels of the sugar glucose in the blood. In type 2 diabetes, there is generally enough insulin but the cells upon it should act are not normally sensitive to its action. The signs and symptoms include increased urine output and decreased appetite as well as fatigue. The major complications of diabetes include dangerously elevated blood sugar, abnormally low blood sugar due to diabetes medications, and disease of the blood vessels which can damage the eyes, kidneys, nerves, and heart.
Metabolic syndrome is a clustering of at least three of five of the following 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 the prevalence in the USA to be an estimated 34% of the adult population, and the prevalence increases with age.
Mother's milk is recommended for all infants for various reasons. Breastfeeding has especially been reported to be beneficial—in comparison to formula feeding—for prevention against later in life health disorders, for example obesity in later 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 risk of type 2 diabetes in later life? A quantitative analysis of published evidence, 2006). It has been widely reported that breast fed infants do have a different growth pattern than infants fed with infant formula. Indeed, breast fed infants have a lower weight gain and a lower body fat mass within the first year of life as compared to infants fed with infant formula. Additionally, breast fed infant have a different gut microbiota profile as compared to infants fed with infant formula. Altogether, these factors affect the development of the infant physiology, including metabolism, immunity and overall growth.
However, in some cases breastfeeding is inadequate or unsuccessful for medical reasons or the mother chooses not to breast feed. Infant formula have been developed for these situations. Fortifiers have also been developed to enrich mother's milk or infant formula with specific ingredients.
Short Chain fatty acids (SCFA) are especially produced by microbial fermentation of dietary fibres in the colon. SCFA, especially butyrate and propionate, have been shown to protect against obesity, insulin resistance, to be involved in adipogenesis, in food intake, in the prevention of non-alcoholic fatty liver disease or of cardiometabolic related conditions like development of 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-producing probiotics reduce nonalcoholic fatty liver disease progression in rats: new insight into the probiotics for the gut-liver axis”, 2013).
SCFA like butyrate and propionate have also been shown to significantly stimulate the release of intestinal hormones like GLP-1 (Chambers et al., 10 Dec. 2014; Hua V. Lin et al, April 2012).
Glucagon-like-peptide-1 (GLP-1 or GLP1) is an incretin secreted by intestinal L-cells upon nutrient intake. Amongst its main effects, it improves glucose clearance by stimulating insulin secretion in pancreatic beta-cells as well as inhibiting glucagon synthesis and secretion simultaneously (Kreymann B et al. 1987, PMID: 2890903; Nathan D M et al. 1992, PMID: 1547685; Nauck M A et al. 1993, PMID: 8423228; Ritzel R et al. 2001, PMID: 11289042). It may therefore prevent type II diabetes.
GLP-1 has also been shown to slow down gastric emptying (Little T J et al. 2006, PMID: 16492694; Nauck M A et al. 1997, PMID: 9374685), to reduce appetite and food intake in both, healthy and obese individuals (Pratley et al. 2008; Orskov et al. 1989; Davis H R 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 A S et al. 2013, PMID: 23380890; Kelly A S et al. 2012, PMID: 22076596). It has also been described that GLP-1 provides some advantageous cardiovascular effects (Bose et al. 2005, PMID: 15616022; Sokos G G et al. 2006, PMID: 17174230).
Increasing colonic SCFA production and/or GLP-1 secretion is therefore an attractive target for preventing later in life health disorders, especially those due to or associated with fat accumulation.
However, orally administered SCFA are unpalatable and are rapidly absorbed in the small intestine. Specific delivery systems targeting the release of propionate in the proximal colon have therefore been developed for some studies. In Chambers et al, scientists have developed a particular carrier molecule whereby propionate is chemically bound by an ester bond to inulin, a natural polymer composed mainly of fructose. This inulin-propionate ester was chemically synthesised. The majority of propionate chemically bound to inulin should only be released when the inulin polymer is fermented by the colonic microbiota, thus providing targeted colonic delivery. However such a kind of carrier present some drawback. This type of chemically synthesized substances may face regulatory issues if used in compositions designed to infants or young children. This study was indeed designed to adults. Some more “natural” solutions, e.g. with ingredients found in breast milk, would therefore be preferred for an administration to infants or young children.
Regarding GLP-1 secretion, two pharmacological approaches have been developed to increase GLP-1 or GLP-1-like activity. The first one is by reducing GLP-1 degradation by inhibiting the enzyme responsible for it (DPP-4i). In addition, several GLP-1 receptor agonists have been used to increase GLP-1 receptor activation. However all these pharmacological approaches are indicated only for adults. More “natural” solutions would be preferred for infants and young children.
Alternative solutions more appropriate to infants and young children should therefore be developed.
As the composition of human milk becomes better understood, it has been proposed to add prebiotics to infant formula. Various infant formulas supplemented with prebiotics such as mixtures of fructo-oligosaccharides (FOS) and galacto-oligosaccharides (GOS) for example are commercially available. Prebiotics are non-digestible in the sense that they are not broken down and absorbed in the stomach or small intestine and thus pass intact to the colon where they are selectively fermented by the bacteria. The main effect of prebiotics, once fermented, is to selectively promote the growth and metabolic activity of certain species of bacteria recognized as beneficial for the host well-being and health (Roberfroid, M, J. Nutrition, 2007: 37(3): 830S-837S). Beyond the direct effects of prebiotic on the gastrointestinal flora, prebiotics are known to also have beneficial effects on the host health (such as anticarcinogenic effects, improvement of mineral absorption and effects on metabolite production) that may be due to indirect effects of the prebiotic on the gut microflora.
Prebiotics such as oligofructose have also been shown to stimulate the GLP-1 gut release (Cani et al., 2005, Phuwamongkolwiwat et al., 2014). However the degree of polymerisation fluctuates widely from a type to another and the induced biological effect can therefore vary greatly.
Moreover, commercially available mixtures approximate only roughly the mixture of oligosaccharides found in human milk. More than 120 different oligosaccharide components have been detected in human milk, some of which have not been detected so far in animal milks (such as bovine milk) at all or have been detected only in small quantities. Some classes of human milk oligosaccharides are present in bovine milk or colostrum only in very small quantities or not at all are sialylated and fucosylated oligosaccharides. As bovine milk contains some oligosaccharides that are structurally identical or similar to those found in human milk, oligosaccharides from bovine milk in sufficient quantities should have prebiotic effect or other beneficial properties associated with human milk oligosaccharides. However until recently, the low concentration of these oligosaccharides in bovine milk (about 20-fold lower than in human milk) has hampered efforts to utilize bovine milk as a source of oligosaccharides for infant formulas.
In addition, few literature exist on the use of bovine milk oligosaccharides for preventing or treating metabolic diseases. The patent application WO2010/003803 describes a nutritional composition for administration to infants which an oligosaccharide mixture consisting of N-acetylated oligosaccharide(s), galacto-oligosaccharide(s) and sialylated oligosaccharide(s) to reduce the risk of obesity later in life. This composition was shown to induce a reduction of triglyceride concentration in the liver with a reduced lipogenesis and triacylglycerol incorporation into lipoproteins in the liver. But no effect of BMOs on the colonic SOFA production and/or on GLP-1 was mentioned or suggested in any of these prior documents.
There is clearly a need for developing new and suitable methods to decrease the risk of later in life health conditions related to excessive fat mass accumulation like obesity or related comorbidities in infants and young children.
There is also a need to deliver such health benefits in a manner that is particularly suitable for these young subjects (infants and young children), in a manner that does not involve a classical pharmaceutical intervention as infants or young children are particularly fragile.
There is a need to deliver such health benefits in infants or young children in a manner that does not induce side effects and/or in a manner that is easy to deliver, and well accepted by the parents or health care practitioners.
There is also a need to deliver such benefits in a manner that does keep the cost of such delivery reasonable and affordable by most.
The present inventors have found that a composition comprising a specific mixture of bovine's milk oligosaccharides (BMOs) can increase colonic SOFA production in an animal model, like propionate, butyrate, valerate and acetate, and especially butyrate and propionate.
They have also found that this specific mixture of bovine's milk oligosaccharides (BMOs) increased secretion of GLP-1 in an in vitro system using human enteroendocrine cells.
These represent new clinical situations where prevention of excessive fat mass accumulation and any related later in life health disorders can be targeted in new ways.
A composition comprising this specific mixture of BMOs can therefore advantageously be used to prevent in an infant or a young child a later in life health disorder related to excessive fat mass accumulation, for example overweight, obesity, insulin resistance, glucose intolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gall bladder disease, cardiovascular disease and metabolic syndrome.
In a particular embodiment, the nutritional composition comprises from 2.5 to 15.0 wt % of the oligosaccharide mixture.
The oligosaccharide mixture comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
It may comprise from 0.1 to 4.0 wt % of the N-acetylated oligosaccharide(s), from 92.0 to 99.5 wt % of the galacto-oligosaccharide(s) and from 0.2 to 4.0 wt % of the sialylated oligosaccharide(s).
The oligosaccharide mixture can be derived from animal milk, such as cow's milk, goat's milk or buffalo's milk.
Abbreviations: Pos ctr=positive control; BMOS=bovine milk oligosaccharides; PDX=polydextrose.
Abbreviations: Pos ctr=positive control; BMOS=bovine milk oligosaccharides; PDX=polydextrose.
Abbreviations: Ctrl pos=positive control; BMOS=bovine milk oligosaccharide; PDX=polydextrose.
Abbreviations: Neg. co=negative control; Pos. co=positive control; BMOS=bovine milk oligosaccharides.
As used herein, the following terms have the following meanings.
The term “infant” means a child under the age of 12 months.
The expression “young child” means a child aged between one and three years, also called toddler.
An “infant or young child born by C-section” means an infant or young child who was delivered by caesarean. It means that the infant or young child was not vaginally delivered.
An “infant or young child vaginally born” means an infant or young child who was vaginally delivered and not delivered by caesarean.
A “preterm” or “premature” means an infant or young child who was not born at term. Generally it refers to an infant or young child born prior 36 weeks of gestation.
The expression “nutritional composition” means a composition which nourishes a subject. This nutritional composition is usually to be taken orally or intravenously. It may include a lipid or fat source, a carbohydrate source and/or a protein source. In a particular embodiment the nutritional composition is a ready-to-drink composition such as a ready-to-drink formula.
In a particular embodiment the composition of the present invention is a hypoallergenic nutritional composition. The expression “hypoallergenic nutritional composition” means a nutritional composition which is unlikely to cause allergic reactions.
In a particular embodiment the nutritional composition of the present invention is a “synthetic nutritional composition”. The expression “synthetic nutritional composition” means a mixture obtained by chemical and/or biological means, which can be chemically identical to the mixture naturally occurring in mammalian milks (i.e. the synthetic nutritional composition is not breast milk).
The expression “infant formula” as used herein refers to a foodstuff intended for particular nutritional use by infants during the first months of life and satisfying by itself the nutritional requirements of this category of person (Article 2(c) of the European Commission Directive 91/321/EEC 2006/141/EC of 22 Dec. 2006 on infant formulae and follow-on formulae). It also refers to a nutritional composition intended for infants and as defined in Codex Alimentarius (Codex STAN 72-1981) and Infant Specialities (incl. Food for Special Medical Purpose). The expression “infant formula” encompasses both “starter infant formula” and “follow-up formula” or “follow-on formula”.
A “follow-up formula” or “follow-on formula” is given from the 6th month onwards. It constitutes the principal liquid element in the progressively diversified diet of this category of person.
The expression “baby food” means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.
The expression “infant cereal composition” means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.
The term “fortifier” refers to liquid or solid nutritional compositions suitable for mixing with breast milk or infant formula.
The expression “weaning period” means the period during which the mother's milk is substituted by other food in the diet of an infant or young child.
The expressions “days/weeks/months/years of life”, “days/weeks/months/years after birth” and “days/weeks/months/years of birth” can be used interchangeably.
The expression “later in life” and “in later life” can be used interchangeably. They refer to effects measured in the individual (infant or young child) after the age of some weeks, some months or some years after birth, such as after the age of 6 months after birth, such as after the age of 8 months after birth, such as after the age of 10 months after birth, such as after the age of 1 year after birth, such as after the age of 2 years, preferably after the age of 4 years, more preferably after the age of 5 years, even more preferably after the age of 7 years after birth, or even more, and as a comparison to average observations for subjects of the same age. Preferably it refers to an effect observed after at least 1 year of life, or after at least 2, 5, 7, 10 or 15 years of life. So the expression “later in life” might refer to an observation during infancy, during early childhood, during childhood, during the adolescent period, or during adulthood. Preferably it refers to an observation during childhood, during the adolescent period, or during adulthood.
The expressions “fat mass accumulation” and “fat accumulation” can be used interchangeably. The expression “excessive fat mass accumulation” refers to abnormal fat mass body amount, e.g. in an amount that can lead to health disorders.
The expressions “reducing excessive fat mass accumulation” and “avoiding excessive fat mass accumulation” refer to a decrease or a limitation of the body fat amount of an individual in order to get a normal or a lower fat mass, e.g. in an amount that does not lead to health disorders.
The expression “health disorder(s)” encompass any health conditions and/or diseases and/or dysfunctions that affect the organism of an individual, including the metabolic ones.
The expressions “preventing a later in life health disorder” or “preventing a health disorder later in life” can be used interchangeably. They mean avoiding that a health disorder/condition occur later in life and/or decreasing the incidence and/or the severity of a health disorder later in life. The prevention occurs “later is life”, so preferably after the termination of the intervention or treatment (i.e. after administration of the nutritional composition according to the invention).
The expression “later in life health disorder related to excessive fat mass accumulation” refers to later in life health disorder due to (so direct link) or associated with (so indirect link) fat excess. It encompasses overweight, obesity and obesity related comorbidities.
“Body mass index” or “BMI” is defined as the value resulting from division of a numerator that is the weight in kilograms by a denominator that is the height in meters, squared. Alternatively, the BMI can be calculated from the weight in pounds as the numerator and the height in inches, squared, as the denominator, with the resultant quotient multiplied by 703. “Overweight” is defined for a human as a BMI between 25 and 30. “Obese” is defined for a human as a BMI greater than 30.
“Obesity related comorbidities” include insulin resistance, glucose intolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gall bladder disease, cardiovascular disease, metabolic syndrome and certain types of cancer.
The term “SCFA” means short chain fatty acid(s). The expression “increasing colonic SCFA production” means that the amount of SCFA, when measured in the colon (or large intestine) or in a part thereof such as the caecum, is higher in an individual fed with the nutritional composition according to the present invention (i.e. comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide) in comparison with a standard composition (i.e. a nutritional composition not comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide) and/or in comparison with a standard composition supplemented with common fibers like polydextrose or pectin. The SCFA may be propionate, butyrate, valerate and/or acetate. In a particular embodiment of the present invention, it is butyrate and/or propionate. The SCFA production may be measured by techniques known by the skilled person such as by 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 intestinal endocrine cells known as L-cells. The expressions “increasing GLP-1 secretion” and “increasing 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 fed with the nutritional composition according to the present invention (i.e. comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide) in comparison with a standard composition (i.e. a nutritional composition not comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide). In a particular embodiment, the expression “increasing GLP-1 secretion” refers to “increasing intestinal GLP-1 secretion”.
The GLP-1 secretion/release may be measured by techniques known by the skilled person such as by measuring its amount in blood circulation (i.e. by determination of plasma GLP-1 concentration) of an individual since GLP-1 is secreted into the bloodstream upon nutrient intake.
The term “growth” refers to growth in weight, height and/or head circumference of an infant or young child. In a particular embodiment it refers to the weight. The growth has to be understood as the evolution of the weight, height and/or head circumference over the aging of the infant or young child. These parameters do not exclusively increase during development of the infant, as indeed the standard curves of growth published by the WHO show that the weight of an infant may decrease in the first days of life of the infant. Therefore, the growth has to be understood as the overall growth of the infant over the first months of life. So the expressions “growth rate” and “rate of growth” can also be used alternatively to the term “growth”.
The expressions “promoting a healthy growth” and “promoting an optimal growth” can be used interchangeably. They encompasses promoting a rate of growth which gets closer or approximates to the rate of growth of a breast-fed infant. They encompass promoting a growth that is qualified as normal by pediatricians so that it is not associated with providing health issues. These expressions also encompass preventing excessive growth or excessive body weight gain that may occur in formula-fed infants, especially in the first few months of life. The expression “promoting a healthy growth” may also encompass controlling weight management and/or avoiding weight gain, especially excessive weight gain, and/or promoting a lean mass increase (especially over a total weight or adipose mass increase).
The “mother's milk” should be understood as the breast milk or the colostrum of the mother.
The term “oligosaccharide” means a carbohydrate having a degree of polymerization (DP) ranging from 2 to 20 inclusive but not including lactose. In some embodiments of the invention, carbohydrate has DP ranging from 3 to 20.
The expressions “at least one N-acetylated oligosaccharide, one galacto-oligosaccharide and one sialylated oligosaccharide” and “at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide” can be used interchangeably.
The expressions “oligosaccharide(s) mixture” or “mixture of oligosaccharide(s)” can be used interchangeably. The oligosaccharide(s) mixture according to the invention comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide. The mixture may be made of one or several oligosaccharides of these different types, i.e. one or several N-acetylated oligosaccharide(s), one or several galacto-oligosaccharide(s) and one or several sialylated oligosaccharide(s). In some advantageous embodiments the oligosaccharides of the oligosaccharide mixture are bovine's milk oligosaccharides (or BMOs).
The expression “N-acetylated oligosaccharide” means an oligosaccharide having N-acetyl residue.
The expressions “galacto-oligosaccharide”, “galactooligosaccharide” and “GOS” can be used interchangeably. They refer to an oligosaccharide comprising two or more galactose molecules which has no charge and no N-acetyl residue (i.e. they are neutral oligosaccharide). 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, “galacto-oligosaccharide” and “GOS” also include oligosaccharides comprising one galactose molecule and one glucose molecule (i.e. disaccharides) which are linked by a β-1,2, β-1,3 or β-1,6 linkage.
The expression “sialylated oligosaccharide” means an oligosaccharide having a sialic acid residue with associated charge.
The terms “prebiotic”, “fibre(s)” and “fiber(s)” can be used interchangeably. They refer to non-digestible carbohydrates that beneficially affect the host by selectively stimulating the growth and/or the activity of healthy bacteria such as bifidobacteria in the colon of humans (Gibson G R, Roberfroid M B. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995; 125:1401-12).
The term “probiotic” means microbial cell preparations or components of microbial cells with a beneficial effect on the health or well-being of the host. (Salminen S, Ouwehand A. Benno Y. et al. “Probiotics: how should they be defined” Trends Food Sci. Technol. 1999:10 107-10). The microbial cells are generally bacteria or yeasts.
The term “cfu” should be understood as colony-forming unit.
All percentages are by weight unless otherwise stated.
The nutritional composition of the present invention can be in solid form (e.g. powder) or in liquid form. The amount of the various ingredients (e.g. the oligosaccharides) can be expressed in g/100 g of composition on a dry weight basis when it is in a solid form, e.g. a powder, or as a concentration in g/L of the composition when it refers to a liquid form (this latter also encompasses liquid composition that may be obtained from a powder after reconstitution in a liquid such as milk, water . . . , e.g. a reconstituted infant formula or follow-on/follow-up formula or infant cereal product or any other formulation designed for infant nutrition).
In addition, in the context of the invention, the terms “comprising” or “comprises” do not exclude other possible elements. The composition of the present invention, including the many embodiments described herein, can comprise, consist of, or consist essentially of the essential elements and limitations of the invention described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise depending on the needs.
Any reference to prior art documents in this specification is not to be considered an admission that such prior art is widely known or forms part of the common general knowledge in the field.
The invention will now be described in further details. It is noted that the various aspects, features, examples and embodiments described in the present application may be compatible and/or combined together.
A first object of the present invention is therefore a nutritional composition comprising an oligosaccharide mixture, said oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide for use in reducing and/or avoiding excessive fat mass accumulation of an infant or a young child by increasing colonic SOFA production and/or by increasing GLP-1 secretion (e.g. intestinal GLP-1 secretion) in said infant or young child. Said excessive fat mass accumulation may be later in life excessive fat mass accumulation in said infant or young child. The nutritional composition of the present invention can also be used for preventing any later in life health disorder related to excessive fat mass accumulation, by increasing colonic SOFA production in an infant or a young child. It can be used for preventing obesity or related comorbidities later in life. Some examples of later in life health disorders related to excessive fat mass accumulation are overweight, obesity, insulin resistance, glucose intolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gall bladder disease, cardiovascular disease and metabolic syndrome.
As illustrated in example 2, the inventors have found that the supplementation of an oligosaccharide mixture according to the present invention increased the SOFA production in an animal model, especially propionate, butyrate, valerate and acetate. Without being bound by theory the inventors of the present invention believe that these particular oligosaccharides act synergically to surprisingly provide a significant increased colonic SOFA production. As illustrated in example 3, the inventors have also found that this specific mixture of bovine's milk oligosaccharides (BMOs) increased secretion of GLP-1 in an in vitro system using human enteroendocrine cells.
Due to the known properties of SOFA, especially butyrate and propionate, and of GLP-1, for example against obesity, insulin resistance, adipogenesis and food intake, such a supplementation of BMOs could therefore be interestingly used to prevent excessive fat mass accumulation and related health disorders.
The oligosaccharide mixture of the nutritional composition according to the invention comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide. As previously mentioned, there may be made of one or several oligosaccharides of these different types, i.e. one or several N-acetylated oligosaccharide(s), one or several galacto-oligosaccharide(s) and one or several sialylated oligosaccharide(s). The oligosaccharide mixture of the nutritional composition of the invention may be prepared from one or more animal milks. The milk may be obtained from any mammal, in particular from cows, goats, buffalos, horses, elephants, camels or sheep.
Alternatively the oligosaccharide mixture may be prepared by purchasing and mixing the individual components.
An N-acetylated oligosaccharide is an oligosaccharide having an N-acetylated residue. Suitable N-acetylated oligosaccharides of the oligosaccharide mixture of the nutritional composition according to the present invention include GalNAcβ1,3Galβ1,4Glc and Galβ1,6GalNAcβ1,3Galβ1,4Glc, but also any mixture thereof. The N-acetylated oligosaccharides may be prepared by the action of glucosaminidase and/or galactoaminidase on N-acetyl-glucose and/or N-acetyl galactose. Equally, N-acetyl-galactosyl transferases and/or N-acetyl-glycosyl transferases may be used for this purpose. The N-acetylated oligosaccharides may also be produced by fermentation technology using respective enzymes (recombinant or natural) and/or microbial fermentation. In the latter case the microbes may either express their natural enzymes and substrates or may be engineered to produce respective substrates and enzymes. Single microbial cultures or mixed cultures may be used. N-acetylated oligosaccharide formation can be initiated by acceptor substrates starting from any degree of polymerization (DP) from DP=1 onwards. Another option is the chemical conversion of keto-hexose (fructose) either free or bound to an oligosaccharide (e.g lactulose) into N-acetylhexosamine or an N-acetylhexosamine containing oligosaccharide as described in Wrodnigg, T. M, Dtutz, A. E, Angew. Chem. Int. Ed. 1999: 38: 827-828.
A galacto-oligosaccharide is an oligosaccharide comprising two or more galactose molecules which has no charge and no N-acetyl residue. Suitable galacto-oligosaccharides 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, but also any mixture thereof. Synthesized galacto-oligosaccharides such as Galβ1,6Galβ1,4Glc, Galβ1,6Galβ1,6Galβ1,6Glc, Galβ1,3Galβ1,4Glc, Galβ1,6Galβ1,6Galβ1,4Glc, Galβ1,6Galβ1,3Galβ1,4Glc, Galβ1,3Galβ1,6Galβ1,4Glc, Galβ1,4Galβ1,4Glc and Galβ1,4Galβ1,4Galβ1,4Glc and mixture thereof are commercially available under trademarks Vivinal® and Elix'or®. Other suppliers of oligosaccharides are Dextra Laboratories, Sigma-Aldrich Chemie GmbH and Kyowa Hakko Kogyo Co., Ltd. Alternatively, specific glycotransferases, such as galoctosyltransferases may be used to produce neutral oligosaccharides.
A sialylated oligosaccharide is an oligosaccharide having a sialic acid residue with associated charge. Suitable sialylated oligosaccharides of the oligosaccharide mixture of the nutritional composition according to the present invention include NeuAcβ2,3Galβ1,4Glc and NeuAcβ2,6Galβ1,4Glc, but also any mixture thereof. These sialylated oligosaccharides may be isolated by chromatographic or filtration technology from a natural source such as animal milks. Alternatively, they may also be produced by biotechnology using specific sialyltransferases either by enzyme based fermentation technology (recombinant or natural enzymes) or by microbial fermentation technology. In the latter case microbes may either express their natural enzymes and substrates or may be engineered to produce respective substrates and enzymes. Single microbial cultures or mixed cultures may be used. Sialyl-oligosaccharide formation can be initiated by acceptor substrates starting from any degree of polymerization (DP) from DP=1 onwards.
In one aspect of the invention, the nutritional composition comprises the oligosaccharide mixture in an amount from 2.5 to 15 wt %. Alternatively, the nutritional composition comprises the oligosaccharide mixture in an amount from 3 to 15 wt %, or in an amount from 3 to 10 wt %, or in an amount from 3.5 to 9.5 wt % or in an amount from 4 to 9 wt % or in an amount from 4.5 to 8.5 wt %, or in an amount from 5.0 to 7.5 wt % such as 5 wt %.
In some specific embodiments, the nutritional composition may comprise the oligosaccharide mixture in an amount from 0.5 to 3.1 g/100 kcal, or in an amount from 0.6 to 3.1 g/100 kcal, or in an amount from 0.6 to 2.0 g/100 kcal, or in an amount from 0.7 to 2.0 g/100 kcal, or in an amount from 0.8 to 1.8 g/100 kcal, or in an amount from 0.9 to 1.7 g/100 kcal, or in an amount from 1.0 to 1.5 g/100 kcal.
The nutritional composition of the present invention may comprise at least 0.01 wt % of N-acetylated oligosaccharide(s), at least 2.0 wt % of galacto-oligosaccharide(s) and at least 0.02 wt % of sialylated oligosaccharide(s).
In some embodiments, the nutritional composition according to the present invention may comprise 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 % of N-acetylated oligosaccharide(s). In some embodiments, it may comprise from 0.005 to 0.06 wt % of N-acetylated oligosaccharide(s) such as 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 % of N-acetylated oligosaccharide(s). A particular example is an amount of 0.01 wt % of N-acetylated oligosaccharide(s).
In addition, the nutritional composition may comprise 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 % of galacto-oligosaccharide(s). In some embodiments, it may comprise from 4.5 to 8 wt % of galacto-oligosaccharide(s) such as from 4.75 to 6 wt % of galacto-oligosaccharide(s) or from 4.9 to 5 wt % or from 5.5 to 6.5 wt % of galacto-oligosaccharide(s). A particular example is an amount of 4.965 wt % of galacto-oligosaccharide(s).
Finally, the nutritional composition may comprise 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 % of sialylated oligosaccharides. In some embodiments, it may comprise from 0.02 to 0.09 wt % of sialylated oligosaccharide(s) such as from 0.02 to 0.07 wt % of sialylated oligosaccharide(s), or from 0.02 to 0.05 wt % of sialylated oligosaccharide(s) or from 0.003 to 0.07 wt % of sialylated oligosaccharide(s). A particular example is an amount of 0.025 wt % of sialylated oligosaccharide(s).
In a particular embodiment, the nutritional composition according to the present invention may comprise from 0.01 to 0.07 wt % of N-acetylated oligosaccharide(s), from 2.0 to 8.0 wt % of galacto-oligosaccharide(s) and from 0.02 to 0.09 wt % of sialylated oligosaccharide(s). In yet another particular embodiment, the nutritional composition according to the present invention may comprise from 0.01 to 0.03 wt % of N-acetylated oligosaccharide(s), 5.95 wt % galacto-oligosaccharide(s) and from 0.02 to 0.09 wt % of sialylated oligosaccharide(s).
In a particular embodiment, the nutritional composition may comprise from 0.0015 to 0.005 g/100 kcal of N-acetylated oligosaccharide(s), from 0.70 to 1.5 g/100 kcal of galacto-oligosaccharide(s) and from 0.0045 to 0.0085 g/100 kcal of sialylated oligosaccharide(s).
In another particular embodiment, the nutritional composition may comprise from 0.0015 to 0.0045 g/100 kcal of N-acetyl-oligosaccharide(s), from 0.74 to 1.2 g/100 kcal of galacto-oligosaccharide(s) and from 0.0045 to 0.0075 g/100 kcal of sialylated oligosaccharide(s).
In a particularly advantageous embodiment, the oligosaccharide mixture of the nutritional composition according to the invention comprises from 0.1 to 4.0 wt % of N-acetylated oligosaccharide(s), from 92.0 to 99.5 wt % of the galacto-oligosaccharide(s) and from 0.2 to 4.0 wt % of the sialylated oligosaccharide(s).
The nutritional composition according to the invention may also contain other types of prebiotic (i.e. different and in addition to the oligosaccharides comprised in the oligosaccharide mixture as defined according to the present invention). Examples of other types of prebiotics include human milk oligosaccharides (HMOs) such as fucosylated oligosaccharides; oligofructose; fructo-oligosaccharides (FOS); inulin; xylooligosaccharides (XOS); polydextrose or any mixture thereof.
A “fucosylated oligosaccharide” is a human milk oligosaccharide having a fucose residue. It has a neutral nature. 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, difucosyllacto-N-hexaose I, difucosyllacto-N-neohexaose II and any combination thereof.
Suitable commercial products that can be used in addition to the oligosaccharides comprised in the oligosaccharide mixture to prepare the nutritional compositions according to the invention include combinations of FOS with inulin such as the product sold by BENEO under the trademark Orafti, or polydextrose sold by Tate & Lyle under the trademark STA-LITE®.
The nutritional composition of the present invention can further comprise at least one probiotic (or probiotic strain), such as a probiotic bacterial strain.
The probiotic microorganisms most commonly used are principally bacteria and yeasts of the following genera: Lactobacillus spp., Streptococcus spp., Enterococcus spp., Bifidobacterium spp. and Saccharomyces spp.
In some particular embodiments, the probiotic is a probiotic bacterial strain. In some specific embodiments, it is particularly Bifidobacteria and/or Lactobacilli.
Suitable probiotic strains include 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 of 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 of New Zealand under the designation K12, Bifidobacterium lactis CNCM I-3446 sold inter alia by the Christian Hansen company of Denmark under the trademark Bb 12, Bifidobacterium longum ATCC BAA-999 sold by Morinaga Milk Industry Co. Ltd. of Japan under the trademark BB536, Bifidobacterium breve sold by Danisco under the trademark Bb-03, Bifidobacterium breve sold by Morinaga under the trade mark M-16V, Bifidobacterium infantis sold by Procter & Gamble Co. under the trademark Bifantis 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 probiotic strain, more preferably between 10e7 and 10e12 cfu such as between 10e8 and 10e10 cfu of probiotic strain per g of composition on a dry weight basis.
In one embodiment the probiotics are viable. In another embodiment the probiotics are non-replicating or inactivated. There may be both viable probiotics and inactivated probiotics in some other embodiments.
The nutritional composition of the invention can further comprise at least one phage (bacteriophage) or a mixture of phages, preferably directed against pathogenic Streptococci, Haemophilus, Moraxella and Staphylococci.
The nutritional composition according to the invention can be for example an infant formula, a starter infant formula, a follow-on or follow-up formula, a baby food, an infant cereal composition, a fortifier such as a human milk fortifier, or a supplement. In some particular embodiments, the composition of the invention is an infant formula, a fortifier or a supplement that may be intended for the first 4 or 6 months of age. 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 breast milk fortifier (e.g. a human milk fortifier) or a formula fortifier such as an infant formula fortifier or a follow-on/follow-up formula fortifier.
When the nutritional composition is a supplement, it can be provided in the form of unit doses.
The nutritional composition of the present invention can be in solid (e.g. powder), liquid or gelatinous form.
The nutritional compositions of the invention, and especially the infant formulas, generally contain a protein source, a carbohydrate source and a lipid source.
The nutritional composition according to the invention generally contains a protein source. The protein can be in an amount of from 1.5 to 3 g per 100 kcal. In some embodiments, especially when the composition is intended for premature infants, the protein amount can be between 2.4 and 4 g/100 kcal or more than 3.6 g/100 kcal. In some other embodiments the protein amount can be below 2.0 g per 100 kcal, e.g. between 1.8 to 2 g/100 kcal, or in an amount below 1.8 g per 100 kcal.
The type of protein is not believed to be critical to the present invention provided that the minimum requirements for essential amino acid content are met and satisfactory growth is ensured. Thus, protein sources based on whey, casein and mixtures thereof may be used as well as protein sources based on soy. As far as whey proteins are concerned, 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.
In some advantageous embodiments the protein source is whey predominant (i.e. more than 50% of proteins are coming from whey proteins, such as 60% or 70%).
The proteins may be intact or hydrolysed or a mixture of intact and hydrolysed proteins. By the term “intact” is meant that the main part of the proteins are intact, i.e. the molecular structure is not altered, for example at least 80% of the proteins are not altered, such as 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, such as at least 98% of the proteins are not altered. In a particular embodiment, 100% of the proteins are not altered.
The term “hydrolysed” means in the context of the present invention a protein which has been hydrolysed or broken down into its component amino acids.
The proteins may be either fully or partially hydrolysed. It may be desirable to supply partially hydrolysed proteins (degree of hydrolysis between 2 and 20%), for example for infants or young children believed to be at risk of developing cow's milk allergy. If hydrolysed proteins are required, the hydrolysis process may be carried out as desired and as is known in the art. For example, whey protein hydrolysates may be prepared by enzymatically hydrolysing the whey fraction in one or more steps. If the whey fraction used as the starting material is substantially lactose free, it is found that the protein suffers much less lysine blockage during the hydrolysis process. This enables the extent of lysine blockage to be reduced from about 15% by weight of total lysine to less than about 10% by weight of lysine; for example about 7% by weight of lysine which greatly improves the nutritional quality of the protein source.
In an embodiment of the invention at least 70% of the proteins are hydrolysed, preferably at least 80% of the proteins are hydrolysed, such as at least 85% of the proteins are hydrolysed, even more preferably at least 90% of the proteins are hydrolysed, such as at least 95% of the proteins are hydrolysed, particularly at least 98% of the proteins are hydrolysed. In a particular embodiment, 100% of the proteins are hydrolysed.
In one particular embodiment the proteins of the nutritional composition are hydrolyzed, fully hydrolyzed or partially hydrolyzed. The degree of hydrolysis (DH) of the protein can be between 8 and 40, or between 20 and 60 or between 20 and 80 or more than 10, 20, 40, 60, 80 or 90.
In a particular embodiment the nutritional composition according to the invention is a hypoallergenic composition. In another particular embodiment the composition according to the invention is a hypoallergenic nutritional composition.
The nutritional composition according to the present invention generally contains a carbohydrate source. This is particularly preferable in the case where the nutritional composition of the invention is an infant formula. In this case, any carbohydrate source conventionally found in infant formulae such as lactose, sucrose, saccharose, maltodextrin, starch and mixtures thereof may be used although one of the preferred sources of carbohydrates is lactose.
The nutritional composition according to the present invention generally contains a source of lipids. This is particularly relevant if the nutritional composition of the invention is an infant formula. In this case, the lipid source may be any lipid or fat which is suitable for use in infant formulae. Some suitable fat sources include palm oil, high oleic sunflower oil and high oleic safflower oil. The essential fatty acids linoleic and α-linolenic acid may also be added, as well small amounts of oils containing high quantities of preformed arachidonic acid and docosahexaenoic acid such as fish oils or microbial oils. The fat source may have a ratio of n-6 to n-3 fatty acids of about 5:1 to about 15:1; for example about 8:1 to about 10:1.
The nutritional composition of the invention may also contain all vitamins and minerals understood to be essential in the daily diet and in nutritionally significant amounts. Minimum requirements have been established for certain vitamins and minerals. Examples of minerals, vitamins and other nutrients 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, phosphorous, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in salt form. The presence and amounts of specific minerals and other vitamins will vary depending on the intended population.
If necessary, the nutritional composition of the invention may contain emulsifiers and stabilisers such as soy, lecithin, citric acid esters of mono- and diglycerides, and the like.
The nutritional composition of the invention may also contain other substances which may have a beneficial effect such as lactoferrin, nucleotides, nucleosides, and the like.
The nutritional composition of the invention may also contain carotenoid(s).
The nutritional composition of the invention (e.g. infant formula) may be prepared by blending together the protein source, the carbohydrate source and the fat source in appropriate proportions. Emulsifiers may be added if desired. Vitamins and minerals may be added at this point but are usually added later to avoid thermal degradation. Any lyophilic vitamins, emulsifiers and the like may be dissolved into the fat source prior to blending. Water, preferably water which has been subjected to reverse osmosis, may then be mixed in to form a liquid mixture.
The liquid mixture may then be thermally treated to reduce bacterial loads. For example, the liquid mixture may be rapidly heated to a temperature in the range of about 80° C. to about 110° C. for about 5 seconds to about 5 minutes. This may be carried out by steam injection or by heat exchanger, e.g. a plate heat exchanger.
The liquid mixture may then by cooled to about 60° C. to about 85° C., for example by flash cooling. The liquid mixture may then be homogenized, for example in two stages at about 7 MPa to about 40 MPa in the first stage and about 2 MPa to about 14 MPa in the second stage.
The homogenized mixture may then be further cooled to add any heat sensitive components such as vitamins and minerals. The pH and solids content of the homogenized mixture may be conveniently standardized at this point.
The homogenized mixture may be transferred to a suitable drying apparatus, such as spray drier or freeze drier, and may be converted to powder. The powder should have a moisture content of less than about 5% by weight.
The oligosaccharide mixture may be prepared by any suitable manner known in the art and added at different steps during the preparation of the nutritional composition of the present invention. The oligosaccharide mixture can be added directly to the nutritional composition (e.g. infant formula) by dry mixing (i.e. at the blending step). Alternatively, the oligosaccharide mixture can be added in liquid mixture prior to the thermal treatment to reduce the bacterial load. The individual components of the oligosaccharide mixture may also be added separately to the nutritional composition in which case the oligosaccharide mixture is preferably added in the liquid phase immediately prior to drying.
The nutritional composition according to the invention is for use in infants or young children. The infants or young children may be born term or preterm. In a particular embodiment the nutritional composition of the invention is for use in infants or young children that were born preterm. In a particular embodiment the nutritional composition of the invention is for use in preterm infants.
The nutritional composition of the present invention may also be used in an infant or a young child that was born by C-section or that was vaginally delivered.
In some embodiments the nutritional composition according to the invention can be for use before and/or during the weaning period.
In some embodiments the nutritional composition according to the invention is for use in infants or young children at risk of developing a later in life health disorder related to excessive fat mass accumulation.
Preterm infants may be at increased risk of insulin resistance, hyperglycemia, poor nutrient utilization, impaired lean body mass growth, fat accumulation in the visceral area and metabolic disease later in life.
Infants or young children at risk of developing later in life overweight or obesity may be targeted.
In some embodiments the nutritional composition of the present invention is for use in infants or young children born from overweight and obese women. Indeed, scientific evidence continues to suggest that infants born to overweight and obese mothers have a greater risk of becoming overweight or obese later in life than infants 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 from mothers who experienced gestational diabetes.
In a particular example, the nutritional composition of the present invention may be used in infants or young children who were IUGR (intrauterine growth restricted). This particular population is at risk of developing a later in life health disorder related to excessive fat mass accumulation since they will have a higher appetite to compensate their growth retardation. But they may not eat in a healthy way with standard formulations, e.g. they may have a higher total weight and/or adipose mass increase over the lean mass increase, which may lead to the development and programming for future health conditions including later in life obesity or future related comorbidities. The nutritional composition of the present invention is believed to provide a healthy growth.
The nutritional composition can be administered (or given or fed) at an age and for a period that depends on the possibilities and needs.
Since the nutritional composition is especially used for prevention purposes (prevention of a later in life health disorder), it can for example be given immediately after birth of the infants. The composition of the invention can also be given during the first week of life of the infant, 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 more. In some particularly advantageous embodiments of the invention, the nutritional composition is given (or administered) to an infant within the first 4 or 6 months of birth of said infant. In some other embodiments, the nutritional composition of the invention is given few days (e.g. 1, 2, 3, 5, 10, 15, 20 . . . ), or few weeks (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 . . . ), or few months (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 . . . ) after birth. This may be especially the case when the infant is premature, but not necessarily.
In one embodiment the composition of the invention is given to the infant or young child as a supplementary composition to the mother's milk. In some embodiments the infant or young child receives the mother's milk during at least the first 2 weeks, first 1, 2, 4, or 6 months. In one embodiment the nutritional composition of the invention is given to the infant or young child after such period of mother's nutrition, or is given together with such period of mother's milk nutrition. In another embodiment the composition is given to the infant or young child as the sole or primary nutritional composition during at least one period of time, e.g. after the 1st, 2nd or 4th month of life, during at least 1, 2, 4 or 6 months.
In one embodiment the nutritional composition of the invention is a complete nutritional composition (fulfilling all or most of the nutritional needs of the subject). In another embodiment the nutrition composition is a supplement or a fortifier intended for example to supplement human milk or to supplement an infant formula or a follow-on formula.
The oligosaccharide mixture present in the nutritional composition of the invention may be prepared from one or more animal milks. The milk can be obtained from any mammal, in particular from cows, goats, buffalos, horses, elephants, camels or sheep. In a specific embodiment, the oligosaccharides of the oligosaccharide mixture are bovine's milk oligosaccharides and can be obtained from cows, goats or buffalos' milk. In an advantageous embodiment, the oligosaccharides are obtained from cow's milk. WO2006087391 and WO2012160080 provide some examples of production of a BMOS mixture.
The present inventors have found that the BMOs intervention in an animal model increased its colonic SOFA production, especially the caecum SOFA production and particularly butyrate and propionate. Since these SOFA have been shown to protect against obesity, insulin resistance, adipogenesis and food intake, the nutritional composition according to the present invention would therefore be useful in reducing and/or avoiding excessive fat mass accumulation in an infant or a young child by increasing colonic SOFA production in said infant or young child.
It can be used to prevent a later in life health disorder related to excessive fat accumulation in an infant or a young child by increasing colonic SOFA production in said infant or young child.
There may be one or several SOFA which production is increased. The SOFA may be propionate, butyrate, valerate and/or acetate. In a particular embodiment the SOFA is propionate and/or butyrate (i.e. propionate, butyrate or both).
Excessive fat accumulation and the related later in life health disorders can be prevented in various ways. Increasing SOFA production represents a new clinical situation where they can be targeted in a new way.
So in a particular embodiment the prevention of excessive fat accumulation and of the related later in life health disorders is obtained by increasing colonic propionate and/or butyrate production in said infant or young child.
In a particular embodiment the propionate and/or butyrate production is measured by Gas-Liquid Chromatography and it can be expressed in nmol/mg dry weight.
In a particular embodiment, the 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% in comparison to the colonic butyrate production obtained with a nutritional composition without at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
In a particular embodiment, the 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, in comparison to the colonic butyrate production obtained with a nutritional composition supplemented with common fibers like polydextrose or pectin.
In a particular embodiment, the 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% in comparison to the colonic propionate production obtained with a nutritional composition without at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
In a particular embodiment, the 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% in comparison to the colonic propionate production obtained with a nutritional composition supplemented with common fibers like polydextrose or pectin.
The present inventors have also found that the BMOs mixture significantly increased the secretion of GLP-1 in an in vitro system using NCI-H716 cells. Since GLP-1 has been especially shown to improve glucose clearance, slow down gastric emptying, reduce appetite and food intake, reduce body weight and provide some advantageous cardiovascular effects, the nutritional composition according to the present invention would therefore be useful in reducing and/or avoiding excessive fat mass accumulation in an infant or a young child. It can be used in an infant or a young child to prevent a later in life health disorder like later in life obesity, diabetes . . . , by increasing GLP-1 secretion (especially the intestinal GLP-1 secretion) in said infant or young child.
In a particular embodiment, the GLP-1 secretion is increased by the BMOs mixture according to 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% in comparison to the GLP-1 secretion obtained with a nutritional composition without said BMOs mixture.
The later in life health disorder that may be prevented by the nutritional composition of the present invention may be any type of health condition associated with excessive fat mass accumulation and for which acting directly or indirectly on the SOFA colonic amount and/or on GLP-1 secretion could have an impact.
As mentioned in the background section, SOFA (especially butyrate and propionate) and GLP-1 are known to protect against obesity, insulin resistance, adipogenesis and food intake. The later in life health disorders related to excessive fat mass accumulation that may be prevented by the nutritional composition of the present invention may be selected from the list consisting of overweight, obesity or obesity related comorbidities. Some examples of later in life obesity related comorbidities are: insulin resistance, glucose intolerance, type 2 diabetes (diabetes mellitus), hypertension, dyslipidemia, sleep apnea, arthritis, hyperuricemia, gall bladder disease, cardiovascular disease, metabolic syndrome and certain types of cancer.
Therefore in some embodiments, the nutritional composition of the present invention can be used for preventing later in life health disorder selected from the 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 later in life obesity.
Another object of the present invention refers to the use of a nutritional composition according to the present invention for reducing the risk of development of overweight later in life in an infant or a young child.
Another object of the present invention refers to the nutritional composition of the present invention for use in promoting a healthy growth in an infant or young child by increasing colonic SOFA production (especially butyrate and propionate) and/or by increasing GLP-1 secretion in said infant or young child, especially to avoid excessive weight gain in said infant or young child.
Another object of the present invention is the use of an oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide in the preparation of a nutritional composition for reducing and/or avoiding excessive fat mass accumulation in an infant or a young child, and/or for preventing a later in life health disorder related to excessive fat mass accumulation in an infant or a young child by increasing colonic SOFA production in 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 an oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide in the preparation of a nutritional composition for promoting a healthy growth in an infant or a young child by increasing colonic SOFA production in 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 an oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide (or the use of a nutritional composition comprising such an oligosaccharide mixture) for increasing colonic SOFA production in an infant or a young child, especially butyrate and/or propionate, and/or for increasing GLP-1 secretion in an infant or a young child.
Another object of the present invention is the use of a nutritional composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide for preventing overweight in an infant or a young child by increasing colonic SOFA production in 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 an oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide for promoting a healthy growth in an infant or a young child by increasing colonic SOFA production in 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 a pharmaceutical composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide for use in reducing and/or avoiding excessive fat mass accumulation in an infant or a young child, and/or for use in preventing a later in life health disorder related to excessive fat accumulation in an infant or a young child by increasing colonic SOFA production in 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 a method for reducing and/or avoiding excessive fat mass accumulation in an infant or a young child, and/or for preventing a later in life health disorder related to fat accumulation in an infant or a young child by increasing colonic SOFA production in said infant or young child, especially butyrate and/or propionate, and/or by increasing GLP-1 secretion in said infant or young child, said method comprising administering to said infant or young child a nutritional composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
Another object of the present invention is a method for promoting a healthy growth in an infant or a young child by increasing colonic SOFA production in said infant or young child, especially butyrate and/or propionate, and/or by increasing GLP-1 secretion in said infant or young child, said method comprising administering to said infant or young child a nutritional composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
Another object of the present invention is a method for increasing colonic SOFA production in an infant or a young child, especially butyrate and/or propionate, and/or for increasing GLP-1 secretion in an infant or a young child, said method comprising administering to said infant or young child a nutritional composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide.
Another object of the present invention is an oligosaccharide mixture comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide for use in a nutritional composition for an infant or a young child as a therapeutic agent that increases colonic SOFA production in said infant or young child, especially butyrate and/or propionate and/or that increases GLP-1 secretion in said infant or young child.
Another object of the present invention is a nutritional composition comprising at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide, for use as a therapeutic agent that increases colonic SOFA production in an infant or a young child, especially butyrate and/or propionate, and/or that increases GLP-1 secretion in an infant or a young child.
The previously-mentioned embodiments and examples (e.g. related to the types and amounts of oligosaccharide, the nutritional composition, the administration, the targeted population . . . ) also apply for all these other objects.
The following examples illustrate some specific embodiments of the composition for use according to the present invention. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible without departing from the spirit of the invention.
An example of the composition of an infant formula comprising an oligosaccharide mixture according to the invention is given in the below table 1. The oligosaccharide mixture may for example comprise from 0.1 to 4.0 wt % of the N-acetylated oligosaccharide(s), from 92.0 to 99.5 wt % of the galacto-oligosaccharide(s) and from 0.2 to 4.0 wt % of the sialylated oligosaccharide(s).
5 week old females BALB/cByJ CRL mice from Charles River were split into several groups and fed during 6 weeks based on the following protocol:
The following fibers were tested:
Table 3 provides a summary of the different tested groups and diets.
After 6 weeks, the animals of each group were sacrificed and the content from caecum was collected. The SOFA production were measured by Gas-Liquid Chromatography (GLC; amounts of SOFA in nmol/mg dry weight). The following SOFA were measured: propionate, butyrate, valerate and acetate.
The measure was made based on the following protocol: SOFA in an acid solution (pH 2.0 to 3.0) were separated on a GLC column coated with a polar stationary phase. This allowed for minimal preparation of the sample (no derivatisation) and straightforward basic FID detection. SCFA were extracted from caecum using an acid phosphate buffer containing HgC12 for inactivation of any residual bacterial activity and an internal standard (2,2 Dimethyl-butyric acid) for GLC analysis. After centrifugation, the sterile-filtered supernatant was ready for analysis by GLC. SOFA were measured simultaneously.
Median ratio values were calculated in order to compare the different fiber-enriched diets on SOFA production.
The production of butyrate by BMOs enriched diet was significantly increased (see
The production of propionate by BMOs enriched diet was also significantly increased (see
These results are very surprising since pectin is usually seen as a high-inducer of SOFA (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).
The inventors therefore surprisingly found that mice fed with a specific BMO mixture were having a higher caecal (and therefore colonic) production for all the tested SOFA, and especially for butyrate and propionate.
Since SOFA, and especially butyrate and propionate, are known to protect against obesity, insulin resistance, adipogenesis, food intake, prevention of non-alcoholic fatty liver disease or of cardiometabolic related conditions like development of atherosclerosis and inflammation, a composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide, would therefore be efficient in infants or young children for use in avoiding excessive fat mass accumulation and in preventing a later in life health disorder associated thereof, by increasing colonic SOFA production in said infants or young children. This represents a new way of targeting prevention of these particular health conditions.
Human NCI-H716 cells were obtained from the American Type Culture Collection. These cells are human enteroendorine cells able to secrete GLP-1.
For proliferation maintenance, the cells were grown in suspension in Gibco® RPMI 1640 supplemented with 10% FBS (Fetal bovine serum), 2 mm I-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin. Cell adhesion and endocrine differentiation were initiated by growing cells in dishes coated with Matrigel in high-glucose DMEM (Dulbecco's Modified Eagle Medium), 10% FBS, 2 mm I-glutamine, 100 IU/ml penicillin, and 100 μg/ml streptomycin.
Two days before the experiments, 1×106 of NCI-H716 cells were seeded in 12-well culture plates coated with Matrigel. On the day of the experiment, supernatants were replaced by KRB buffer (Krebs-Ringer Bicarbonate Buffer) with or without the tested agent:
Cells were incubated for 2 h at 37° C. with the different effectors. The 2 h cell incubation with the different effectors did not affect cell viability. Supernatants were collected with the addition of Complete EDTA-Free Protease inhibitor cocktail, DPP-IV (Dipeptidyl peptidase 4) inhibitor and frozen at −80° C. for subsequent analysis. Cells were scraped off and sonicated in a homogenization analysis. Cells were scraped off and sonicated for normalization. To normalize GLP-1 content, cell homogenate proteins were measured. Biologically active GLP-1 was measured by enzyme-linked immunosorbent assay (ELISA) as described by the distributor of GLP-1 (active) ELISA kit (EMD Millipore; # EGLP-35K).
The secretion of GLP-1 was significantly increased with the addition of BMOS in this in vitro system using NCI-H716 human cells (P<0.01 vs Negative control; two-sided t-test). It was especially increased by 92% in comparison to the negative control.
Since GLP-1 is known to improve glucose clearance, slow down gastric emptying, reduce appetite and food intake, reduce body weight and provide some advantageous cardiovascular effects, a composition comprising an oligosaccharide mixture that comprises at least one N-acetylated oligosaccharide, at least one galacto-oligosaccharide and at least one sialylated oligosaccharide, would therefore be efficient in infants or young children for use in avoiding excessive fat mass accumulation and in preventing a later in life health disorder like obesity and diabetes, by increasing GLP-1 secretion in said infants or young children. This represents a new way of targeting prevention of these particular health conditions.
| Number | Date | Country | Kind |
|---|---|---|---|
| 16152751.0 | Jan 2016 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2017/051592 | 1/26/2017 | WO | 00 |
