FIELD OF THE INVENTION

The present invention relates to a nutritional composition for infants at risk of a disadvantageous body composition development for example due to being born to a mother that developed gestational diabetes, for use in improving body composition of such infants.

BACKGROUND OF THE INVENTION

Human milk is the uncontested gold standard concerning infant nutrition. However, in some cases breastfeeding is inadequate or unsuccessful for medical reasons or because of a choice not to breastfeed. For such situations infant or follow-on formulas have been developed. Commercial infant formulas are commonly used today to provide supplemental or sole source of nutrition early in life. These formulas comprise a range of nutrients to meet the nutritional needs of the growing infant, and typically include fat, carbohydrate, protein, vitamins, minerals, and other nutrients helpful for optimal infant growth and development. Commercial infant formulas are designed to mimic, as closely as possible, the composition and function of human milk.

Human milk lipids are known to have a distinct physical structure composed of large lipid globules with an average mode diameter of about 4 pm existing of a triglyceride core coated by a tri-layer of membranes, the milk fat globule membrane (MFGM). The mode diameter of lipid droplets in standard infant formula is typically about 0.3-0.5 pm due to the industrial processing procedures to achieve stable and reproducible end products, and the lipid droplets are not surrounded by MFGM but mostly by milk proteins. Infant formula with lipid globules with an architecture more similar to the lipid globules in human milk have been described.

Gestational diabetes mellitus (GDM) is a serious health condition that arises during pregnancy. It is characterized by hyperglycemia during pregnancy and results in short- and long-term consequences for the mother and child. Notably, infants born from a GDM pregnancy have an increased risk of overweight and obesity in infancy, childhood and in later life (Logan et al Arch Dis Child Fetal Neonatal Ed 2017;102:F65-F72; Farahvar et al Expert Review of Endocrinology & Metabolism, 2018, 14(1):63-74; Nijs and Benhalima J Clin Med 2020, 9, 599; doi:10.3390/ jcm9020599). Current treatment options are limited and focus mainly on diet and lifestyle interventions or insulin therapy for the pregnant women, yet these options do not appear to decrease the longer term risks such as developing a disadvantageous body composition, in particular with regard to fat mass and lean body mass, for their children.

WO 2013/191533 relates to infant formulas comprising vegetable fat and mammalian milk fat and having large lipid globules that preferably also have phospholipids in the outer layer of the lipid globules for the prevention of obesity later in life and the improvement of body composition. These infants were born from healthy mothers and were preterm/small for gestational age that often encounter catch up growth early in life.

Similarly, US 2016/0081963 describes an infant formula for infants born from healthy mothers to prevent/reduce obesity later in life wherein the infant formula comprises at least 10 wt. % palmitic acid based on total fatty acids, wherein at least 30 wt. % of the palmitic acid is esterified to the sn- 2 position of a triglyceride based on total palmitic acid.

US 2016/219915 involves a composition comprising large lipid globules containing vegetable fat as the lipid ingredient which are coated with phospholipids and polar lipids for improving body composition later in life in infants born from a mother with diabetes/obesity.

Non pre-published WO 2021/110916 describes that infants born from overweight or obese mothers receiving an infant formula comprising a mix of vegetable fat and milk fat improves the growth trajectory or body development in terms of Body Mass Index (BMI).

SUMMARY OF THE INVENTION

The present inventors have surprisingly found that the consumption of a nutritional composition comprising a mixture of vegetable fat and milk fat and having large lipid globules coated with phospholipids beneficially affects the body composition in subjects at risk of developing a disadvantageous body composition.

A mouse model was used, wherein a GDM-like phenotype (gestational diabetes mellitus) was induced in dams by feeding them a high-fat diet (to reduce insulin sensitivity) and exposing them to three mild streptozotocin triggers (to reduce beta cell capacity). The offspring of these GDM dams were fed in early life either with a nutritional composition according to the invention, i.e. a nutritional composition comprising a mixture of vegetable fat and milk fat and comprising large lipid globules coated with phospholipids, or a control nutritional composition comprising mainly vegetable fat with small lipid globules without a phospholipid coating. Offspring of GDM dams are more at risk of overweight and obesity and thus developing a disadvantageous body composition. After exposure to a Western style diet (WSD) later in life, the body weight of the offspring fed with either nutritional composition early in life were not significantly different. However, the offspring fed the nutritional composition according to the invention developed an unexpected greater increase in lean body mass compared to the offspring that was fed the control nutritional composition. This is indicative that consumption in early life of the nutritional composition according to the invention improves body composition in infants at risk of developing a disadvantageous body composition

Since the overall fatty acid composition in the diets was very similar, the observed much higher gain in lean body mass was wholly unexpected, all the more since the diet later in life for both groups was the same obesogenic Western style diet. Thus, due to the presence in the early life diet of lipid containing a mixture of vegetable fat and milk fat and because the lipid was in the form of large globules coated with phospholipids, this early life diet was able to program differences in body composition development which results in a healthier body composition later in life, i.e. adulthood. The present invention hence relates to nutritional compositions, in particular formulae for infants or growing up milks for toddlers, comprising a mixture of vegetable fat and milk fat and wherein the lipid is in the form of large lipid globules comprising a coating that comprises phospholipids that can be used to improve body composition in infants at risk of developing a disadvantageous body composition for example due to being born to a mother that developed gestational diabetes.

DETAILED DESCRIPTION OF THE INVENTION

The present invention thus pertains to a nutritional composition selected from infant formula, follow- on formula and young child formula, comprising digestible carbohydrates, protein and lipid, wherein the lipid comprises i) 30 to 90 wt% vegetable lipid based on total lipid, ii) 10 to 70 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat wherein the lipid is in the form of lipid globules and wherein a) the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and/or at least 45 volume %, based on total lipid volume, of the lipid globules have a diameter of

2 to 12 pm; and b) the lipid globules are at least partly coated on the surface with phospholipids, the amount of phospholipids present in the nutritional composition being from 0.5 to 20 wt% phospholipids based on total lipid for use in improving body composition in a human subject selected from the group consisting of an infant born to an overweight and/or obese mother at the time of conception of the infant; an infant born to a mother with diabetes at the time of conception of the infant; - an infant born to a mother with gestational diabetes (GDM); and an infant who was large for its gestational age (LGA) at birth.

For some jurisdictions, the invention can also be worded as the use of digestible carbohydrates, protein and lipid in the manufacture of a nutritional composition, selected from infant formula, follow- on formula and young child formula, for improving body composition in a human subject selected from the group consisting of an infant born to an overweight and/or obese mother at the time of conception of the infant; an infant born to a mother with diabetes at the time of conception of the infant; - an infant born to a mother with gestational diabetes (GDM); and an infant who was large for its gestational age (LGA) at birth wherein the lipid comprises i) 30 to 90 wt% vegetable lipid based on total lipid, ii) 10 to 70 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat wherein the lipid is in the form of lipid globules and wherein a) the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and/or at least 45 volume %, based on total lipid volume, of the lipid globules have a diameter of 2 to 12 pm; and b) the lipid globules are at least partly coated on the surface with phospholipids, the amount of phospholipids present in the nutritional composition being from 0.5 to 20 wt% phospholipids based on total lipid.

For some jurisdictions, the invention can also be worded as a method for improving body composition in a human subject selected from the group consisting of an infant born to an overweight and/or obese mother at the time of conception of the infant; an infant born to a mother with diabetes at the time of conception of the infant; an infant born to a mother with gestational diabetes (GDM); and - an infant who was large for its gestational age (LGA) at birth said method comprising feeding said human subject a nutritional composition, selected from infant formula, follow-on formula and young child formula, said nutritional composition comprising digestible carbohydrates, protein and lipid, wherein the lipid comprises i) 30 to 90 wt% vegetable lipid based on total lipid, ii) 10 to 70 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat wherein the lipid is in the form of lipid globules and wherein a) the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and/or at least 45 volume %, based on total lipid volume, of the lipid globules have a diameter of 2 to 12 pm; b) and the lipid globules are at least partly coated on the surface with phospholipids, the amount of phospholipids present in the nutritional composition being from 0.5 to 20 wt% phospholipids based on total lipid. In some jurisdictions administering a nutritional composition to an infant is considered non- therapeutic. In those instances the invention can be worded as defined above by way of a method comprising administering a nutritional composition. For clarity, the method can also be defined as a non-therapeutic method for improving body composition in an infant as defined above. By definition, the words “non-therapeutic” exclude any therapeutic effect. In the context of the present invention, the infant formula or follow-on formula is not native cow’s milk or human milk.

In the context of the present invention, young child formula can also be named growing-up milk.

In the context of the present invention a mother that is overweight or obese at the time of conception of the infant is based on the BMI of the mother before pregnancy. In one embodiment, the BMI before pregnancy is defined as the BMI as measured in the time period at least six months before conception.

In this specification, the following terms have the meanings assigned to them here below (Chiu M et al. Diabetes Care, 2011 , 34:1741-1748):

The term ‘BMI’ as used herein, is defined as the body mass divided by the square of the body height, and is expressed in units of kg/m ² . The BMI broadly categorizes a person as underweight, normal weight, overweight, or obese according to the calculated BMI value.

The terms ‘underweight’, ‘normal weight’, ‘overweight’ and ‘obese’ as herein are defined as follows: Underweight is defined as a BMI < 18.5 kg/m2 for all women. A normal weight for Asian women is considered a BMI >18.5 and <23 kg/m2, for non-Asian women, a normal weight is a BMI >18.5 and

<25 kg/m2. Overweight for Asian women is considered a BMI > 23 and < 27.5 kg/m2, for non-Asian women, overweight is a BMI > 25 and <30 kg/m2. Obese for Asian women is considered a BMI > 27.5 kg/m2, for non-Asian women, obese is a BMI > 30 kg/m2. In the context of the present invention a ‘mother with diabetes at the time of conception’ is a mother who has been diagnosed by a health care professional with diabetes mellitus type 1 or diabetes mellitus type 2. In the context of this invention this diagnosis of diabetes mellitus type 1 or 2 was made by a health care professional at the latest at the end of the 1 ^st trimester during the pregnancy of the infant. Preferably the mother with diabetes at the time of conception is a mother with diabetes mellitus type 2.

In the context of the present invention a ‘mother with gestational diabetes’ is a mother who has been diagnosed by a health care professional to suffer from gestational diabetes mellitus (GDM) during pregnancy. In the context of the present invention the GDM diagnosis is preferably based on IADPSG criteria (IADPSG: International Association of Diabetes and Pregnancy Study Group). Typically, a GDM diagnosis is made at the earliest in the second trimester during pregnancy. GDM occurs in pregnant women without a previous history of diabetes and is transient, i.e. the pathology disappears when the woman is not pregnant anymore. In the context of the present invention ‘an infant who was large for its gestational age (LGA) at birth’ is an infant with a weight, length, or head circumference at birth that lies above the 90th percentile for that gestational age. Both infants born to women with pre-pregnancy overweight or obesity and infants born to women with diabetes (diabetes mellitus type 1 , diabetes mellitus type 2 or GDM) have a higher risk of being LGA.

The unifying concept between these sub-groups is that all these infants are or have a higher risk of being exposed in utero to increased levels of glucose compared to infants born from healthy mothers. Exposure to increased levels of glucose or hyperglycaemia lead to both short- as well as long-term effects in the infant. The long-term effects may comprise increased risk of developing obesity and/or cardiovascular diseases in later life and neurodevelopmental problems in early life. The most predominant effect is the development of a disadvantageous body composition with regard to an increased or higher than average fat mass. Throughout this description, lipid and fat are used interchangeably.

Application

According to the present invention, the human subject is selected from the group consisting of an infant born to an overweight and/or obese mother at the time of conception of the infant; - an infant born to a mother with diabetes at the time of conception of the infant; an infant born to a mother with GDM; and an infant who was LGA at birth.

The infants listed here above are all considered by the skilled person to be populations at-risk of developing a disadvantageous body composition later in life.

In a preferred embodiment the human subject is selected from the group consisting of an infant born to an overweight and/or obese mother at the time of conception of the infant; an infant born to a mother with diabetes at the time of conception of the infant; and an infant born to a mother with GDM.

In a more preferred embodiment the human subject is selected from the group consisting of: an infant born to a mother with diabetes at the time of conception of the infant; and an infant born to a mother with GDM. In an even more preferred embodiment the human subject is an infant born to a mother with GDM.

In an even more preferred embodiment the human subject is an infant born to an overweight and/or obese mother at the time of conception of the infant and wherein the mother also has GDM. According to the present invention the body composition of the above defined ‘at risk’ infants is improved. The improvement in body composition is compared to the body composition of the same ‘at risk’ infants who did not consume the nutritional composition comprising a mixture of vegetable fat and milk fat and having large lipid globules coated with phospholipids but instead consumed a standard or control nutritional composition. The standard or control nutritional composition is characterised by comprising at least 95 wt% vegetable lipid based on total lipid and less than 2 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat and less than 0.5 wt% phospholipids based on total lipid and having lipid globules with a mode diameter, based on volume, of about 0.3-0.5 pm.

Thus in a preferred embodiment according to the invention, improving body composition is compared to the body composition of a human subject fed a nutritional composition, selected from infant formula, follow-on formula and young child formula, comprising digestible carbohydrates, protein and lipid, said nutritional composition comprising at least 95 wt% vegetable lipid based on total lipid and less than 1 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat and less than 0.5 wt% phospholipids based on total lipid and having lipid globules with a mode diameter, based on volume, of about 0.3- 0.5 pm. For a proper comparison, the standard or control nutritional composition is fed to a human subject from the same group and is of the same age and is fed for the same period of time.

In a preferred embodiment according to the invention, improving body composition is selected from the group consisting of increasing lean body mass and reducing the percentage of fat mass, preferably improving body composition is increasing lean body mass. In this embodiment, improving body composition is not the same as improving body development or growth trajectory in which weight is compared to length (BMI). It may still be that infants having average weights versus average lengths and consequently an average BMI nevertheless have an increased fat mass or a lowered lean body mass and thus a disadvantageous body composition. Hence, in a preferred embodiment, improving body composition means, importantly, that the ratio of fat mass to lean mass is reduced.

In yet a further preferred embodiment according to the invention, improving body composition is increasing gain in lean body mass. Preferably, the percentage of lean mass is increased by at least 5%, more preferably by at least 8%, even more preferably the percentage of lean body mass is increased by at least 10%, most preferably by at least 12% compared to subjects given an infant milk formula (IMF) comprising predominantly vegetable fat._Preferably the gain in the increase in lean body mass is established after the administration of the nutritional composition has stopped. In other words, preferably the gain in the increase in lean body mass is established later in life. In a preferred embodiment, the human subject is exposed to a Western style diet later in life. A Western-style diet is known in the art to be a general unhealthy diet, characterized by a large content of (unhealthy) lipids and a large content of rapidly digestible carbohydrates, in particular sucrose. A Western-style diet may also be referred to as a “high-fat diet”, as a “cafeteria diet” or as “unhealthy diet”. Typically, a Western-style diet is high in fat and high in saturated fat. In the context of the present invention, the Western-style diet is defined as containing more than 45 % lipid, based on total calories of the daily food intake, wherein the lipid contains at least 50 % by weight of saturated fats, such as present in for example cream, cheese, butter, ghee, suet, tallow, lard, and fatty meats. Further, the Western-style diet is defined as containing more than 50 % carbohydrate, based on total calories of the daily food intake, wherein the carbohydrate contains at least 75 % by weight of rapidly digestible carbohydrates selected from the group consisting of glucose, fructose, sucrose, lactose and starch.

In a preferred embodiment, the present invention concerns a nutritional composition selected from infant formula, follow-on formula and young child formula, comprising digestible carbohydrates, protein and lipid, wherein the lipid comprises i) 30 to 90 wt% vegetable lipid based on total lipid, ii) 10 to 70 wt% based on total lipid of mammalian milk lipid derived from the group consisting of butter, butter fat, butter oil and anhydrous milk fat wherein the lipid is in the form of lipid globules and wherein a) the lipid globules have a mode diameter, based on volume, of at least 1 .0 pm and/or at least 45 volume %, based on total lipid volume, of the lipid globules have a diameter of 2 to 12 pm; and b) the lipid globules are at least partly coated on the surface with phospholipids, the amount of phospholipids present in the nutritional composition being from 0.5 to 20 wt% phospholipids based on total lipid for use in increasing gain in lean body mass later in life in an infant born to a mother with gestational diabetes (GDM), preferably wherein the infant is exposed to a Western style diet. Lioid

The nutritional composition for use according to the present invention comprises lipid. Lipid in the present invention comprises one or more selected from the group consisting of triglycerides, polar lipids (such as phospholipids, cholesterol, glycolipids, sphingomyelin), free fatty acids, monoglycerides and diglycerides. Preferably the composition comprises at least 70 wt%, more preferably at least 80 wt%, more preferably at least 85 wt% triglycerides, even more preferably at least 90 wt% triglycerides based on total lipid.

The lipid provides preferably 30 to 60 % of the total calories of the nutritional composition. More preferably the nutritional composition comprises lipid providing 35 to 55 % of the total calories, even more preferably the nutritional composition comprises lipid providing 40 to 50 % of the total calories. The lipid is preferably present in an amount of 3 to 7 g per 100 kcal, more preferably in an amount of 4 to 6 g lipid per 100 kcal and most preferably in an amount of 4.5 to 5.5 g lipid per 100 kcal. When in liquid form, e.g. as a ready-to-feed liquid, the nutritional composition preferably comprises 2.1 to 6.5 g lipid per 100 ml, more preferably 3.0 to 4.0 g per 100 ml. Based on dry weight the nutritional composition preferably comprises 10 to 50 wt%, more preferably 12.5 to 40 wt% lipid, even more preferably 19 to 30 wt% lipid.

The lipid comprises vegetable lipid. The presence of vegetable lipid advantageously enables an optimal fatty acid profile high in polyunsaturated fatty acids and/or more reminiscent to human milk fat. Lipid from non-human mammalian milk alone, e.g. cow’s milk, does not provide an optimal fatty acid profile. The amount of essential fatty acids is too low in non-human mammalian milk.

Preferably the nutritional composition comprises at least one, preferably at least two vegetable lipid sources selected from the group consisting of linseed oil (flaxseed oil), rape seed oil (such as colza oil, low erucic acid rape seed oil and canola oil), sunflower oil, high oleic sunflower oil, safflower oil, high oleic safflower oil, olive oil, coconut oil, palm oil and palm kernel oil.

In a preferred embodiment, the nutritional composition comprises 30 to 90 wt% vegetable lipid based on total lipid, more preferably 35 to 80 wt%, more preferably 40 to 70 wt%, more preferably 40 to 60 wt% vegetable lipid based on total lipid.

The lipid in the nutritional composition for the use according to the invention further comprises lipid from mammalian milk, preferably ruminants milk, preferably cow's milk, goat milk, sheep milk, buffalo milk, yak milk, reindeer milk, and camel milk, most preferably cow's milk. Preferably the mammalian milk is not human milk. Preferably the mammalian milk component comprises at least 70 wt% triglycerides, more preferably at least 90 wt., more preferably at least 97 wt%.

Preferably the mammalian milk lipid is derived from the group consisting of butter, butter fat, butter oil, and anhydrous milk fat, more preferably anhydrous milk fat and butter oil. Such milk fat lipid sources are high in triglyceride levels. Furthermore these lipid sources are in the form of a continuous fat phase or a water-in-oil emulsion form. Using these sources of milk fat during the manufacture of the nutritional composition of the present invention enable the formation of lipid globules, wherein each globule comprising a mixture of vegetable fat and milk fat. When milk fat sources are used which are an oil-in-water emulsion, lipid globules being either composed of milk fat or composed of vegetable fat will be generated, which are believed to be less effective.

Milk fat in the present invention refers to all lipid components of milk, as produced by the mammalians, such as the cow, and is found in commercial milk and milk-derived products. Butter in the present invention is a water-in-oil emulsion comprised of over 80 wt% milk fat. Butterfat in the present invention relates to all of the fat components in milk that are separable by churning, in other words, present in butter. Anhydrous milk fat (AMF) is a term known in the art and relates to extracted milk fat. Typically AMF comprises more than 99 wt% lipid based on total weight. It can be prepared from extracting milk fat from cream or butter. Anhydrous butter oil in the present invention is synonymous with AMF.

Butteroil also is a term known in the art. It typically relates to a milk lipid extract with more than 98 wt% lipid and typically is a precursor in the process of preparing anhydrous milk fat or anhydrous butter oil.

Preferably the composition comprises 10 to 70 wt% milk lipid based on total lipid, more preferably 20 to 65 wt%, more preferably 30 to 60 wt%, more preferably 40 to 60 wt% based on total lipid. Preferably this milk lipid is selected from the group consisting of butter, butter fat, butter oil, and anhydrous milk fat.

Preferably the ratio of vegetable fat to milk fat ranges from 3/7 to 9/1 . In a preferred embodiment, the lipid in the nutritional composition comprises: a) 35 to 80 wt% vegetable lipid based on total lipid, and b) 20 to 65 wt% mammalian milk fat based on total lipid, wherein the mammalian milk fat is selected from butter, butter fat, butter oil or anhydrous milk fat.

More preferably, the lipid in the nutritional composition comprises: a) 40 to 70 wt% vegetable lipid based on total lipid, and b) 30 to 60 wt% mammalian milk fat based on total lipid, wherein the mammalian milk fat is selected from butter, butter fat, butter oil or anhydrous milk fat.

Most preferably, the lipid in the nutritional composition comprises: a) 40 to 60 wt% vegetable lipid based on total lipid, and b) 40 to 60 wt% mammalian milk fat based on total lipid, wherein the mammalian milk fat is selected from butter, butter fat, butter oil or anhydrous milk fat.

The nutritional composition also may comprise non-vegetable lipid and non-milk fat, such as animal fat other than milk fat, such as fish oil, and egg lipid, and microbial, algal, fungal or single cell oils. Preferably the non-vegetable, non-milk fat is present in an amount of at most 10 wt% based on total lipid, more preferably at most 5 wt%. Preferably the lipid in the nutritional composition comprises a fat source comprising long chain poly-unsaturated fatty acids (LC-PUFA), selected from the group consisting of fish oil, marine oil, algal oil, microbial oil, single cell oil and egg lipid in an amount of 0.25 to 10 wt% based on total lipid, preferably in an amount of 0.5 to 10 wt%. Compared to vegetable fat, milk fat is known to have a higher content of palmitic acid (PA) at the sn-2 position of a triglyceride. In a preferred embodiment, the lipid in the nutritional composition for use according to the invention comprises at least 10 wt% palmitic acid (PA) based on total fatty acids and at least 15 wt% of palmitic acid, based on total palmitic acid, is located at the sn-2 position of a triglyceride. Preferably the amount of PA that is present is below 30 wt% based on total fatty acids. More preferably the amount of PA that is present in the lipid is from 12 to 26 wt% based on total fatty acids in the total lipid, even more preferably from 14 to 24 wt%.

The lipid in the nutritional composition is preferably chosen such that, based on the total PA present in the lipid, at least 15 wt%, preferably at least 20 wt%, more preferably at least 25 wt%, more preferably at least 30 wt% PA is in the sn-2 or beta position in a triglyceride. Preferably the amount of PA in the sn-2 position in a triglyceride is not more than 45 wt%, preferably not more than 40 wt% based on total PA present in the lipid. Preferably the amount of PA in the sn-2 position in a triglyceride is from 25 to 40 wt% based on total PA present in the total lipid.

Compared to vegetable fat, milk fat is known to have a higher content of short-chain fatty acids butyric acid (BA; C4) and caproic acid (CA; C6). In a preferred embodiment, the lipid in the nutritional composition for use according to the invention comprises 0.6 to 5 wt% short chain fatty acids (SCFA) being the sum of butyric acid (BA) and caproic acid (CA) based on total fatty acids. Preferably the nutritional composition comprises less than 5 wt% BA based on total fatty acids, preferably less than 4 wt%. Preferably the nutritional composition comprises at least 0.5 wt% butyric acid based on total fatty acids, preferably at least 0.6 wt%, preferably at least 0.9 wt%, more preferably at least 1 .2 wt% BA based on total fatty acids. In a preferred embodiment, the lipid in the nutritional composition for use according to the invention comprises at least 10 wt% palmitic acid based on total fatty acids and at least 15 wt% of palmitic acid, based on total palmitic acid, is located at the sn-2 position of a triglyceride and comprises 0.6 to 5 wt% short chain fatty acids (SCFA) being the sum of butyric acid (BA) and caproic acid (CA) based on total fatty acids.

Faty acid composition

SFA relates to saturated fatty acids and/or acyl chains, MUFA relates to mono-unsaturated fatty acid and/or acyl chains, PUFA refers to polyunsaturated fatty acids and/or acyl chains with 2 or more unsaturated bonds; LC-PUFA refers to long chain polyunsaturated fatty acids and/or acyl chains comprising at least 20 carbon atoms in the fatty acyl chain and with 2 or more unsaturated bonds; Medium chain fatty acids (MCFA) refer to fatty acids and/or acyl chains with a chain length of 6, 8 or 10 carbon atoms. n3 or omega-3 PUFA refers to polyunsaturated fatty acids and/or acyl chains with 2 or more unsaturated bonds with an unsaturated bond at the third carbon atom from the methyl end of the fatty acyl chain, n6 or omega-6 PUFA refers to polyunsaturated fatty acids and/or acyl chains with 2 or more unsaturated bonds with an unsaturated bond at the sixth carbon atom from the methyl end of the fatty acyl chain.

DHA refers to docosahexaenoic acid and/or acyl chain (22:6, n3); DPA refers to docosapentaenoic acid and/or acyl chain (22:5 n3). EPA refers to eicosapentaenoic acid and/or acyl chain (20:5 n3); ARA refers to arachidonic acid and/or acyl chain (20:4 n6). LA refers to linoleic acid and/or acyl chain (18:2 n6); ALA refers to alpha-linolenic acid and/or acyl chain (18:3 n3). PA relates to palmitic acid and/or acyl chains (C16:0). BA refers to butyric acid (C4:0). CA refers to caproic acid (C6:0). LA refers to linoleic acid and/or acyl chain and is an n6 PUFA (18:2 n6) and the precursor of n6 LC- PUFA and is an essential fatty acid as it cannot be synthesized by the human body. The nutritional composition according to the present use preferably comprises linoleic acid (LA). LA preferably is present in a sufficient amount in order to promote a healthy growth and development, yet in an amount as low as possible to prevent negative, competitive, effects on the formation of n3 PUFA and a too high n6/n3 ratio. The nutritional composition therefore preferably comprises less than 20 wt% LA based on total fatty acids, preferably 5 to 16 wt%, more preferably 10 to 14.5 wt%. Preferably, the nutritional composition comprises at least 5 wt% LA based on total fatty acids, preferably at least 6 wt% LA, more preferably at least 7 wt% LA based on total fatty acids. Per 100 kcal, the nutritional composition preferably comprises 350 - 1400 mg LA.

ALA refers to a-linolenic acid and/or acyl chain and is an n3 PUFA (18:3 n3) and the precursor of n3 LC-PUFA and is an essential fatty acid as it cannot be synthesized by the human body. The nutritional composition according to the present use preferably comprises ALA. Preferably ALA is present in a sufficient amount to promote a healthy growth and development of the infant. The nutritional composition therefore preferably comprises at least 1.0 wt%, more preferably the nutritional composition comprises at least 1.5 wt%, even more preferably at least 2.0 wt% ALA based on total fatty acids. Preferably the nutritional composition comprises less than 10 wt% ALA, more preferably less than 5.0 wt% based on total fatty acids. Preferably the nutritional composition comprises a weight ratio of LA/ALA from 2 to 20, more preferably from 3 to 16, more preferably from 4 to 14, more preferably from 5 to 12.

The lipid that is present in the nutritional composition for use according to the invention preferably comprises 5 to 35 wt% poly-unsaturated fatty acids (PUFA), based on total fatty acids, comprising linoleic acid (LA) and alpha-linolenic acid (ALA) in a weight ratio LA/ALA of 2 to 20.

Preferably, the nutritional composition comprises n3 LC-PUFA, such as EPA, DPA and/or DHA, more preferably DHA. As the conversion of ALA to DHA may be less efficient in infants, preferably both ALA and DHA are present in the nutritional composition. Preferably the nutritional composition comprises at least 0.05 wt%, preferably at least 0.1 wt%, more preferably at least 0.2 wt%, of DHA based on total fatty acids. Preferably the nutritional composition comprises not more than 2.0 wt%, preferably not more than 1 .0 wt% of DHA based on total fatty acids.

The nutritional composition in one embodiment comprises ARA. Preferably the nutritional composition comprises at least 0.05 wt%, preferably at least 0.1 wt%, more preferably at least 0.2 wt% of ARA based on total fatty acids. As the group of n6 fatty acids, especially arachidonic acid (ARA) counteracts the group of n3 fatty acids, especially DHA, the nutritional composition preferably comprises relatively low amounts of ARA. Preferably the nutritional composition comprises not more than 2.0 wt%, preferably not more than 1.0 wt% of ARA based on total fatty acids. Preferably the weight ratio between DHA and ARA is between 1 :4 to 4:1 , more preferably between 1 :2 to 2:1 , more preferably between 0.6 and 1 .5. ARA may also be absent.

Lipid globule size

According to the present invention, lipid is present in the nutritional composition in the form of lipid globules. When the nutritional composition is in liquid form, these lipid globules are emulsified in the aqueous phase. Alternatively, when the nutritional composition is in powder form, the lipid globules are present in the powder and the powder is suitable for reconstitution with water or another food grade aqueous phase. The lipid globules comprise a core and a surface. The lipid globules in the nutritional composition preferably have mode diameter, based on volume, of at least 1 .0 pm, more preferably at least 3.0 pm, and most preferably at least 4.0 pm. Preferably, the lipid globules have a mode diameter, based on volume, between 1 .0 and 10 pm, more preferably between 2.0 and 8.0 pm, even more preferably between 3.0 and 7.0 pm, and most preferably between 4.0 pm and 6.0 pm.

Alternatively, or preferably in addition, the size distribution of the lipid globules is preferably in such a way that at least 45 volume % (vol%), preferably at least 55 vol%, even more preferably at least 65 vol%, and most preferably at least 75 vol% of the lipid globules have a diameter between 2 and 12 pm. In a preferred embodiment, at least 45 vol%, preferably at least 55 vol%, more preferably at least 65 vol%, and most preferably at least 75 vol% of the lipid globules have a diameter between 2 and 10 pm. In a more preferred embodiment, at least 45 vol%, more preferably at least 55 vol%, yet even more preferably at least 65 vol%, and most preferably at least 75 vol% of the lipid globules have a diameter between 4 and 10 pm. Preferably less than 5 vol% of the lipid globules have a diameter above 12 pm.

Standard infant formulas, follow-on formulas or young child formulas have lipid globules with a mode diameter, based on volume, of about 0.3-0.5 pm. The volume percentage of lipid globules is based on volume of total lipid. The mode diameter relates to the diameter which is the most present based on volume of total lipid, or the peak value in a graphic representation, having on the X-as the diameter and on the Y-as the volume %. The volume of the lipid globule and its size distribution can suitably be determined using a particle size analyzer such as a Mastersizer (Malvern Instruments, Malvern, UK), for example by the method described in Michalski et al, 2001 , Lait 81 : 787-796.

Phospholipid The nutritional composition for use according to the present invention comprises 0.5 to 20 wt% phospholipid based on total lipid, more preferably 0.5 to 10 wt%, more preferably 0.75 to 8 wt%, even more preferably 1.0 to 8 wt%, most preferably 1.5 to 5 wt% phospholipid based on total lipid.

Phospholipids are amphipathic of nature and include glycerophospholipids and sphingomyelin. By ‘coating’ is meant that the outer surface layer of the lipid globules comprises phospholipid, whereas phospholipid is virtually absent in the core of the lipid globule. A suitable way to determine whether phospholipid is located on the surface of lipid globules is confocal laser scanning microscopy or transmission electron microscopy; see for instance Gallier et al. (A novel infant milk formula concept: Mimicking the human milk fat globule structure, Colloids and Surfaces B: Biointerfaces 136 (2015) 329-339).

The nutritional composition preferably comprises glycerophospholipids. Examples of glycerophospholipids are phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylglycerol (PG). Preferably the nutritional composition comprises one or more of PC, PS, PI and PE, more preferably the nutritional composition comprises at least PC.

The nutritional composition preferably comprises sphingomyelin. Sphingomyelins have a phosphorylcholine or phosphorylethanolamine molecule esterified to the 1 -hydroxy group of a ceramide. They are classified as phospholipid as well as sphingolipid, but are not classified as a glycerophospholipid nor as a glycosphingolipid. Preferably the nutritional composition comprises 0.05 to 10 wt% sphingomyelin based on total lipid, more preferably 0.1 to 5 wt%, even more preferably 0.2 to 2 wt%. Preferably the nutritional composition comprises at least 5 wt%, more preferably 5 to 40 wt% sphingomyelin based on total phospholipid, more preferably 10 to 35 wt%, even more preferably 15 to 35 wt%, based on total phospholipid.

The nutritional composition preferably comprises glycosphingolipids. Preferably the nutritional composition comprises 0.1 to 10 wt% glycosphingolipids based on total lipid, more preferably 0.5 to 5 wt%, even more preferably 2 to 4 wt%, based on total lipid. The term glycosphingolipids in the present context particularly refers to glycolipids with an amino alcohol sphingosine. The sphingosine backbone is O-linked to a charged head-group such as ethanolamine, serine or choline backbone. The backbone is also amide linked to a fatty acyl group. Glycosphingolipids are ceramides with one or more sugar residues joined in a beta-glycosidic linkage at the 1 -hydroxyl position, and include gangliosides. Preferably the nutritional composition contains gangliosides, more preferably at least one ganglioside selected from the group consisting of GM3 and GD3.

The nutritional composition preferably comprises phospholipid derived from mammalian milk. Preferably the nutritional composition comprises phospholipid and glycosphingolipid derived from mammalian milk. The nutritional composition preferably comprises phospholipid and optionally glycosphingolipid from mammalian milk from cows, mares, sheep, goats, buffalos, horses and/or camels. More preferably the nutritional composition comprises phospholipid and optionally glycosphingolipid from cow’s milk.

Phospholipid derived from milk includes preferably phospholipid that is isolated from milk lipid, cream lipid, cream serum lipid, butter serum lipid (beta serum lipid), whey lipid, cheese lipid and/or buttermilk lipid. Buttermilk lipid is typically obtained during the manufacture of buttermilk. Butter serum lipid or beta serum lipid is typically obtained during the manufacture of anhydrous milk fat from butter. Preferably the phospholipid and optionally glycosphingolipid is obtained from milk cream. Examples of suitable commercially available sources for phospholipid from milk are BAEF, SM2, SM3 and SM4 powder of Corman, Salibra of Glanbia, Lipamin M20 of Lecico and LacProdan MFGM-10 or PL20 of Aria.

The use of phospholipid from milk lipid advantageously comprises the use of milk fat globule membranes, which are more reminiscent to the situation in human milk. The concomitant use of phospholipid derived from milk and triglycerides derived from vegetable lipid and mammalian milk fat therefore enables the manufacture of coated lipid globules with a coating more similar to human milk, while at the same time providing an optimal fatty acid profile.

Preferably the phospholipid is derived from milk lipid, more preferably from cow’s milk lipid. Preferably the phospholipid is derived from orforms part of the milkfat globule membrane (MFGM), more preferably is derived from or forms part of cow’s MFGM.

Preferably the nutritional composition comprises phospholipid and glycosphingolipid. In a preferred embodiment the weight ratio of phospholipid : glycosphingolipid is from 2:1 to 12:1 , more preferably from 2:1 to 10:1 and even more preferably 2:1 to 5:1 .

Methods for obtaining lipid globules with an increased size and coating with phospholipid are for example disclosed in WO 2010/0027258 and WO 2010/0027259. Digestible carbohydrates

The nutritional composition comprises digestible carbohydrates. The digestible carbohydrates preferably provide 25 to 75% of the total calories of the nutritional composition. Preferably the digestible carbohydrates provide 40 to 60% of the total calories. Based on calories the nutritional composition preferably comprises of 5 to 20 g of digestible carbohydrates per 100 kcal, more preferably 6 to 16 g per 100 kcal. When in liquid form, e.g. as a ready-to-feed liquid, the nutritional composition preferably comprises 3 to 30 g digestible carbohydrate per 100 ml, more preferably 6 to 20, even more preferably 7 to 10 g per 100 ml. Based on dry weight the nutritional composition preferably comprises 20 to 80 wt%, more preferably 40 to 65 wt% digestible carbohydrates.

Preferred digestible carbohydrate sources are lactose, glucose, sucrose, fructose, galactose, maltose, starch and maltodextrin. Lactose is the main digestible carbohydrate present in human milk. Lactose advantageously has a low glycaemic index. The nutritional composition preferably comprises lactose. The nutritional composition preferably comprises digestible carbohydrate, wherein at least 35 wt%, more preferably at least 50 wt%, more preferably at least 75 wt%, even more preferably at least 90 wt%, most preferably at least 95 wt% of the digestible carbohydrate is lactose. Based on dry weight the nutritional composition preferably comprises at least 25 wt% lactose, preferably at least 40 wt% lactose. Protein

The nutritional composition comprises protein. The protein preferably provides 5 to 20% of the total calories. Preferably the nutritional composition comprises protein that provides 6 to 12% of the total calories. Preferably the nutritional composition comprises less than 3.5 g protein per 100 kcal, more preferably the nutritional composition comprises between 1.5 and 2.1 g protein per 100 kcal, even more preferably between 1.6 and 2.0 g protein per 100 kcal. A low protein concentration advantageously is closer to human milk as human milk comprises a lower amount of protein based on total calories compared to cow’s milk. The protein concentration in a nutritional composition is determined by the sum of protein, peptides and free amino acids. Based on dry weight the nutritional composition preferably comprises less than 12 wt% protein, more preferably between 9.6 and 12 wt%, even more preferably between 10 and 11 wt%. Based on a ready-to-drink liquid product the nutritional composition preferably comprises less than 1.5 g protein per 100 ml, more preferably between 1 .2 and 1 .5 g, even more preferably between 1 .25 and 1 .35 g per 100 ml.

The source of the protein should be selected in such a way that the minimum requirements for essential amino acid content are met and satisfactory growth is ensured. Hence protein sources based on cows' milk proteins such as whey, casein and mixtures thereof and proteins based on soy, potato or pea are preferred. In case whey proteins are used, the protein source is preferably based on acid whey or sweet whey, whey protein isolate or mixtures thereof. Preferably the nutritional composition comprises at least 3 wt% casein based on dry weight. Preferably the casein is intact and/or non-hydrolyzed. Non-diaestible carbohydrates

In one embodiment the nutritional composition preferably comprises non-digestible oligosaccharides. Preferably the nutritional composition comprises non-digestible oligosaccharides with a degree of polymerization (DP) between 2 and 250, more preferably between 3 and 60.

Preferably the nutritional composition comprises fructo-oligosaccharides, galacto-oligosaccharides and/or galacturonic acid oligosaccharides, more preferably fructo-oligosaccharides and/or galacto- oligosaccharides, even more preferably galacto-oligosaccharides, most preferably transgalacto- oligosaccharides. In a preferred embodiment the nutritional composition comprises a mixture of galacto-oligosaccharides and fructo-oligosaccharides, more preferably transgalacto- oligosaccharides and fructo-oligosaccharides. Suitable non-digestible oligosaccharides are for example Vivinal®GOS (FrieslandCampina DOMO), Raftilin®HP or Raftilose® (Orafti). Preferably, the nutritional composition comprises 80 mg to 2 g non-digestible oligosaccharides per 100 ml, more preferably 150 mg to 1.5 g, even more preferably 300 mg to 1 g per 100 ml. Based on dry weight, the nutritional composition preferably comprises 0.25 wt% to 20 wt%, more preferably 0.5 wt% to 10 wt%, even more preferably 1.5 wt% to 7.5 wt% of non-digestible oligosaccharides. Formula

The use according to the present invention requires the administration of an infant formula, a follow- on formula or a young child formula. This means that the composition that is administered is not human milk. It also means that the composition that is administered is not native cow’s milk or native milk from another mammal. Alternatively, the terms as used herein, “infant formula” or “follow-on formula” or “young child formula” means that it concerns a composition that is artificially made or in other words that it is synthetic. Hence in one embodiment, the nutritional composition that is administered is an artificial infant formula or an artificial follow-on formula or an artificial young child formula or a synthetic infant formula or a synthetic follow-on formula or a synthetic young child formula.

In the present context, infant formula refers to nutritional compositions, artificially made, intended for infants of 0 to about 4 to 6 months of age and are intended as a substitute for human milk. Typically, infant formulae are suitable to be used as sole source of nutrition. Such infant formulae are also known as starter formula. Follow-on formula for infants starting with at 4 to 6 months of life to 12 months of life are intended to be supplementary feedings to infants that start weaning on other foods. Infant formulae and follow-on formulae are subject to strict regulations, for example for the EU regulations no. 609/2013 and no. 2016/127. In the present context, young child formula refers to nutritional compositions, artificially made, intended for infants of 12 months to 36 months, which are intended to be supplementary feedings to infants. The nutritional composition is preferably an infant formula or a follow-on formula. More preferably the nutritional composition is an infant formula.

The nutritional composition for use according to the present invention comprises digestible carbohydrates, protein and lipid, wherein preferably the lipid provides 30 to 60 % of the total calories, the protein provides 5 to 20% of the total calories and the digestible carbohydrates provide 25 to 75% of the total calories.

The nutritional composition is preferably an infant formula or follow-on formula and preferably comprises 3 to 7 g lipid/100 kcal, preferably 4 to 6 g lipid/100 kcal, more preferably 4.5 to 5.5 g lipid/100 kcal, preferably comprises 1.7 to 3.5 g protein/100 kcal, more preferably 1.8 to 2.1 g protein/100 kcal, more preferably 1.8 to 2.0 g protein/100 kcal and preferably comprises 5 to 20 g digestible carbohydrate/100 kcal, preferably 6 to 16 g digestible carbohydrate/100 kcal, more preferably 10 to 15 g digestible carbohydrate/100 kcal.

Preferably the nutritional composition is an infant formula or follow-on formula, and preferably has an energy density of 60 kcal to 75 kcal/100 ml, more preferably 60 to 70 kcal/100 ml, when in a ready-to-drink form. This density ensures an optimal balance between hydration and caloric intake. In one embodiment, the nutritional composition is a powder. Suitably, the nutritional composition is in a powdered form, which can be reconstituted with water or other food grade aqueous liquid, to form a ready-to drink liquid, or is in a liquid concentrate form that should be diluted with water to a ready-to-drink liquid. It was found that lipid globules maintained their size and coating when reconstituted.

In this document and in its claims, the verb "to comprise" and its conjugations is used in its nonlimiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one".

EXAMPLES

Example 1: Effect of lipid globule design on lean body mass in offspring of dams with gestational diabetes

An experiment was performed wherein the effects of an infant milk formula (IMF) comprising a mixture of vegetable fat and milk fat with large phospholipid coated lipid globules (IMF 1) was compared with a control IMF based on vegetable fat with small lipid globules and no phospholipid coating (IMF 2). GDM mouse model

A gestational diabetes mellitus (GDM) mouse model was used to generate offspring which was exposed to GDM conditions during gestation. The dams received 2 triggers before pregnancy in order to induce a transient GDM condition. This approach reflected the GDM pathophysiology in a pilot study (Li et al. Journal of Endocrinology 2020, 244:501-510).

At 12 weeks of age C57BL/6N dams were fed with a 60 energy % high fat diet (HFD) for 4 weeks to reduce insulin sensitivity. The HFD was based on AIN93G diet with an adjusted lipid fraction containing 60 energy % lipid. A low dose of Streptozotocin (60 mg STZ/kg) was administered on 3 consecutive days prior to mating. STZ is an alkylating agent that selectively kills pancreatic beta- cells via cell necrosis and/or apoptosis. By administrating low doses of STZ, the beta-cell capacity is reduced. The offspring was breastfed/lactated with breast milk till PN16. At PN16, the dam and litters were with IMF 1 (n=10) or the control IMF 2 (n=11). The male offspring were weaned at P21 and continued on the same IMF diet (1 or 2) until PN42. The offspring was subsequently fed with Western-style diet (WSD) for 8 weeks. The WSD was based on AIN93G diet with an adjusted lipid fraction containing 45 energy% lipid.

At PN42 and PN98, the male offspring were assessed for body weight (using standard scale) as well as fat mass and lean mass, using an MRI analyser (EchoMRMOOH, EchoMRI) per manufacturer’s instructions.

Diets

The offspring diets comprised a macronutrient and micronutrient composition following AIN93G. The offspring diets consisted of 28.3% (w/w) IMF 1 or control IMF 2. Protein, carbohydrates, and micronutrients were added to match AIN93G. The fat components were derived entirely from the IMF. The fat content and fatty acid profile of the diets comprising IMF 1 and IMF 2 were similar (Table 1).

For IMF 1 a mix of anhydrous cow’s milk fat, coconut oil, low erucic acid rape seed oil, sunflower oil, high oleic acid sunflower oil, with a small amount of soy lecithin and LC-PUFA premix was used. In addition, IMF 1 comprised butter serum powder as a source of milk derived phospholipids. The amount of vegetable lipid was about 49 wt% based on total fat, and the amount of mammalian milk fat about 50 wt%. For control IMF 2 a mixture of palm oil, coconut oil, low erucic acid rape seed oil, sunflower oil, high oleic sunflower oil, with a small amount of soy lecithin, and LC-PUFA premix was used. The amount of vegetable lipid in the final control IMF was about 98 wt% based on total fat, and the amount of mammalian milk fat about 1 wt%. Table 1 : Characteristics of the IMF powder

IMF 1 was prepared in a similar way as described in example 1B of WO 2010/0027259. The lipid globules of IMF 1 were large and coated with phospholipids. The control IMF 2 was prepared with high pressure homogenization, resulting in small lipid globules.

The detailed characteristics of the fat component of the different experimental IMFs used for the different IMFs are shown in Table 2. Table 2 Results

At PN98 the body weight and lean body mass were measured. Table 3 shows the body weight and lean body mass at PN98, as well as the percentage change in lean body mass from PN42 - PN98 Table 3

* p-value < 0.05 IMF 1 vs. IMF 2 ** p-value = 0.053 IMF 1 vs. IMF 2

As can be deduced from Table 3, whilst there was no difference in body weight at PN98, the levels of lean body mass in mice which consumed IMF 1 in early life and subsequently have been exposed to a WSD diets were significantly higher compared to the body mass in the mice that consumed IMF 2 in early life. The mice exposed to IMF 1 in early life also gained more lean mass during the exposure to the WSD (PN42 - PN98) compared to the mice exposed to IMF 2 in early life. Increased lean body mass after exposure to a WSD is considered healthier and is indicative of better metabolic handling of the dietary challenge.

Since the overall fatty acid composition in the diets was very similar, it is surprising that the presence of milk fat and difference in lipid globule design (larger lipid globules with PL coating) in an IMF fed early in life has such a programming effect towards investment in lean body mass during the WSD challenge (PN42 to PN98).

Example 2 Infant formula:

Infant formula, intended for infants of 0 to 6 months of age, comprising per 100 ml, after reconstituting 13.7 g powder to an end volume of 100 ml: - 66 kcal

1 .3 g protein (whey protein/casein weight ratio 1/1)

7.3 g digestible carbohydrates (mainly being lactose)

3.4 gram fat (of which about 47 wt% bovine milk fat, about 1 .5 wt% buttermilk powder as a source of milk derived phospholipids, the remainder being vegetable oils, fish oil and microbial oil)

0.8 g non-digestible oligosaccharides, of which 0.08 g long chain fructo-oligosaccharides (source RaftilineHP) and 0.72 g trans-galacto-oligosaccharides (source Vivinal GOS) minerals, vitamins, trace elements and other micronutrients as according to directives for infant formula. The formula comprises lipid globules with a volume mode diameter of about 5.6 pm and the volume % of lipid globules with a mode diameter between 2 and 12 pm is above 45.