Composition comprising glycosphingolipids for young child applications

Abstract

Composition comprising Glycosphingolipids for young child applications This invention relates to compositions comprising one or more glycosphingolipids, to child nutritional formulas which contain said compositions, to uses of said compositions for supporting healthy growth and development of young children, in particular, infants and toddlers, and dietary management of symptoms associated with inflammation of the gastrointestinal system of said young children. In particular, the invention relates to a composition comprising one or more compounds of formula (1), or salts thereof: wherein R1 is a C13-C17 alkyl chain; R2 is a C15-C19 alkyl chain; and R3 is a glycosyl moiety selected from Neu5Acα2-3-Galβ1-4-Glcβ1-, or Neu5Acα2-8-Neu5Acα2-3-Galβ1-4-Glc β1-. Child nutritional formulas containing the composition are also provided.

Description

[0001] Description

[0002] Composition comprising Glycosphingolipids for young child applications

[0003] Filed of the Invention

[0004] This invention relates to compositions comprising one or more glycosphingolipids, to child nutritional formulas which contain said compositions, to uses of said compositions for supporting healthy growth and development of young children, in particular, infants and toddlers, and dietary management of symptoms associated with inflammation of the gastrointestinal system of said young children.

[0005] Background of the Invention

[0006] Breast feeding is recommended for all infants as human milk contains all the essential nutrients for optimal growth and development of the infant. Human milk is broadly composed of water, carbohydrates, lipids, proteins, vitamins, and minerals. The lipid fraction of human milk occurs mostly as triacylglycerols which are dispersed throughout the milk in globules. The globules are encapsulated by a tri-layer membrane called the milk fat globule membrane. The triacylglycerol fraction provides about 98% of milk lipids in human milk and delivers more than 50% of the energy intake of the new-born infant.

[0007] The remaining 2% of milk lipids includes components of the milk fat globule membrane such as polar lipids, cholesterol (including free and cholesteryl esters), mono- and di-glycerides, and free fatty acids. Although a relatively minor fraction in terms of abundance, the milk fat globule membrane component is nutritionally important by serving as a source of cholesterol and polar lipids. Cholesterol is believed to aid early brain development of the neonate. The polar lipids in milk are glycerophospholipids and sphingolipids such as sphingomyelin and gangliosides. The glycerophospholipids serve as essential components of the lipid bilayer in all cell membranes and are important in various biological processes, including lipid and cholesterol digestion, absorption and transport, and regulation of plasma and liver lipid and lipoprotein levels. Sphingomyelin is a vital component of the brain myelination process and plays an active role in regulation of inflammation and in signaling pathways.

[0008] The gangliosides are thought to positively impact gastrointestinal and immune health by two main mechanisms: influencing the gut microbiota to promote the growth of bifidobacteria, and by binding to pathogenic targets in the intestine to reduce infections. Gangliosides have also been shown to influence the expression of gut tight junction proteins, maintaining intestinal integrity during episodes of gut inflammation (Park et al, 2010, Journal of Pediatric Gastroenterology and Nutrition, 50, 321). It is also believed that the gangliosides play a critical role in the development of the infant's central nervous system. The most abundant ganglioside in mature human milk is GM3, which has one sialic acid side-unit in its carbohydrate chain. The ganglioside GD3, which has two sialic acid units, is also present at relatively high levels in early-lactation milk but levels diminish to low levels with time.

[0009] Unfortunately, breastfeeding is not always possible or practicable, and therefore there is a need for human milk substitutes. These substitutes usually take the form of starter infant formulas for newborn infants and follow-on formulas for infants of 6 to 12 months of age. Also, for infants having special needs, certain medical formulas are used. These formulas are typically produced using reduced-fat dairy products such as bovine skim milk, or fractions obtained from bovine skim milk. This means that, at best, these formulas contain levels of polar lipids and cholesterol which are significantly lower than the levels in human milk. However, many of these formulas are formulated with vegetable oils rather than bovine lipids. As a result, the milk fat present in many formulas is typically very low, although supplementation of milk fat is becoming more prevalent. Where formula is supplemented with bovine milk fat, it is often by using anhydrous milk fat, which contains cholesterol but not polar lipids. Even when bovine milk fats are used, bovine milk contains significantly lower levels of gangliosides and human milk. Therefore, when reconstituted, the resulting formulas still contain much less gangliosides than human milk.

[0010] The infant nutrition industry has responded by incorporating bovine milk fat globule membrane fractions into infant formula (EP2211629). This enables the overall ganglioside levels of infant formulas to be brought closer to those in human milk. However, issues remain. In bovine milk, ganglioside GD3 is much more abundant than ganglioside GM3. As mentioned above, human milk contains greater amounts of ganglioside GM3. The difference in the ratio of GM3 to GD3 in human and bovine milk is likely due to their different biological and developmental requirements (Pan et al, 2000, Early Human Development 57, 25-31).

[0011] Further, GM3 and GD3 are not single compounds but are collective names for groups of compounds with similar structures. In particular, the fatty acids attached to the sphingoid base of the ganglioside can vary in chain length. The fatty acid chain lengths in human GM3 and GD3 are predominantly of 20 or less carbon atoms, predominantly of 16 and 18 carbon atoms, while those in bovine milk are predominantly of 22 or more carbon atoms (Bode et al, 2004, J. Nutr. 134: 3016-3020; Martin-Sosa et al, 2004, Lipids 39, 111-116). This difference has biological relevance. The fatty acid composition of gangliosides influences membrane fluidity and alters the formation of sphingolipid clusters in the cell membrane, which is important for cell- cell interactions, receptor-ligand interactions, and signaling pathways. Many infants suffer from inflammation of the gastro-intestinal tract. This negatively impacts the integrity and functioning of the gut epithelial barrier. A proper functioning gut epithelial barrier is crucial for the overall wellbeing of the infant. The inflammation can extend from sub-chronic to chronic to acute. Depending upon the severity of the inflammation, there can be various consequences for the infant ranging from inadequate nutritional intake, colic, pain, and increased risk to develop diseases later in life. These diseases include intestinal bowel disease, allergies, and autoimmune conditions, amongst others.

[0012] The microbiota of the human intestine, which is a complex and very dynamic microbial ecosystem, is considered to serve numerous important functions for its human host, including protection against pathogens and induction of immune regulatory functions. The intestinal microbiota consists of various populations, which are important to preserve human health, and recent research has been able to link imbalances in the intestinal bacterial population to both intestinal and extra-intestinal inflammatory diseases.

[0013] Selective stimulation of specific intestinal bacteria to promote their growth and metabolic activity could be a helpful approach in creating a benign intestinal microbial community. Particularly, this may be achieved by exploiting the capability of specific intestinal bacteria to grow on carbon sources which may be less easily used by other members of the intestinal microbial community.

[0014] Bifidobacteria are considered one of the most beneficial probiotics. As an example, strains of B.bifidum and B. longum have been widely studied for their immunomodulatory properties and protecting effect against pathogens (Chichlowski et al. J. Pediatr. Gastroenterol. Nutr. 2012, 55, 321).

[0015] A probiotic attribute of bifidobacteria is the direct or indirect production of short chain fatty acids (SCFAs), such as acetate and butyrate, which largely contribute to intestinal functionality.

[0016] Particularly, it has been shown that acetate produced by bifidobacteria acts in vivo to promote defense functions of the host epithelial cells and thereby protects the host from infection (Gut Microbes 2012, 3, 449-54).

[0017] In addition, while bifidobacteria do not directly produce butyrate as fermentation end-product, the importance of metabolic cross-feeding on acetate by butyrate-producing bacteria in the gut has been demonstrated. A study has shown that the butyrate-producing species such as f. ha / lii can utilize intermediates of bifidobacterial fermentation, such as acetate and lactate, and produce butyrate (Schwab et al. Frontiers in Microbiology 2017, 8, 1).

[0018] Butyrate is the primary energy source for colonocytes and has been reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene and tight junction expression. Additionally, butyrate has immunomodulatory effects keeping the immune cells in balance. Several studies have shown a protective effect of butyrate which can reduce the risk of disease including irritable bowel syndrome (IBS), and inflammatory bowel disease (IBD) (Canani et al. World J. Gastroenterol. 2017, 17, 1519).

[0019] Certainly, there is a need for means, preferably orally administered means, more preferably dietary means, to effectively increase the concentration of SCFAs in the gastro-intestinal tracts of humans, especially of infants.

[0020] Summary of the invention

[0021] In a first aspect, this invention relates to a composition comprising one or more compounds of formula (1), or salts thereof: wherein

[0022] R1is a C13-C17 alkyl chain;

[0023] R2is a C15-C19 alkyl chain; and

[0024] R3is a glycosyl moiety selected from Neu5Aca2-3-Gaipi-4-Glcpi-, or Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-.

[0025] In another aspect, this invention relates to a child nutritional formula which comprises the composition defined above.

[0026] In another aspect, this invention provides a child nutritional formula to treat a healthy young child to prevent or reduce the risk of occurrence of an inflammatory condition, or to treat a pediatric patient to reduce the symptoms of inflammation in that pediatric patient, e.g. a condition and the symptoms of colic, allergy, food- or pathogen-induced inflammation in the gastrointestinal tract (GIT), necrotizing enterocolitis (NEC) or very early onset-inflammatory bowel disease (VEO-IBD). In one aspect, this invention provides a child nutritional formula for reducing bacterial pathogen -induced inflammation, such as lipopolysaccharide induced inflammation, in the gastro-intestinal tract in an infant or a young child.

[0027] Various embodiments relating to these aspects of the invention are described throughout this specification and are illustrated by non-limiting examples. Description of the Figures

[0028] Figure 1. Schematic outline of the study of the impact on metabolite production in the gut of healthy infants stimulated by compositions of the present invention described in Example 7.

[0029] Detailed Description of the Invention

[0030] It has been surprisingly found that it is possible to produce, at scale and at reasonable cost, a composition comprising gangliosides identical to those present in human milk. Further, the composition can contain other glycosphingolipids, such as lactosyl ceramide and / or glucosyl ceramide, which are identical to those present in human milk. This enables the provision of child nutritional formulas which are closer in composition to human milk. By comprising gangliosides which are identical in structure to gangliosides in human milk, the composition is better tolerated by the infant or young child. Further, it has been surprisingly found that a composition which comprising synthetic gangliosides identical to those found in human milk stimulates the production of shortchain fatty acids (SCFA), in particular, butyrate, in gastrointestinal tract (GIT )of young children, and is effective in reduction of inflammation in such young children. One of the outcomes of the increased production of SCFAs, is a decrease in the pH of the colonic lumen, which in turn increases mineral absorption and inhibits growth of pathogens. Increased levels of SCFAs, such as acetate and butyrate, in the GIT are proven to be associated with a series of potential health benefits for humans, especially young children, like reduction of inflammation and allergic diseases, improved integrity of the GIT epithelium and nutrient absorption, reduced pathogen-induced inflammation (Zhang, P. Int. J. Mol. Sci. 2022, 23: 9588; Zhang, P. Nutrients 2023, 15: 3683; Silva YP et al Front Endocrinol 2020, 11; Vinolo MAR et al Nutrients. 2011 3: 858)

[0031] This invention is described in general terms that relate to all aspects and embodiments and have the following meaning unless otherwise specified:

[0032] "About" or "approximately" mean being close to the value or range following these terms, as understood by one of ordinary skill in the art and include a deviation up to 10 % of the values or ranges that follow.

[0033] "Young child" in the present context means a young human of age up to about 3 years, such as an infant or a toddler. "Infant" means a child of age up to 12 months old. "Toddler" means is a child of age between 12 to 36 months old.

[0034] "Child nutritional formula" means a foodstuff which is suitable as a complete or partial substitute for human milk and is intended to satisfy the particular nutritional needs of infants during the first year of life, or children of ages about 1 year to about 3 years. The child nutritional formula may be (i) a starter infant formula (also called an infant formula) which is suitable for infants from birth, (ii) a follow on formula which is suitable for infants over the age of six months, (iii) a food for special medical purposes / medical food which is intended for infants who have special dietary needs and who are unable to consume standard starter infant formulas and / or follow-on formulas, or (iv) a growing up milk which is suitable for supplementing the diet of young children of ages about 1 to about 3 years. Infant formulas and follow-on infant formulas are regulated in Europe under EU Regulation 2016 / 127. Food for special medical purposes for infants are regulated in Europe under EU Regulation 2016 / 128. Equivalent regulations exist in other geographies.

[0035] "Comprising" means including the compound of interest and other related or non-related compounds. Related terms such as "contain", "contained", "containing", "comprise" and "comprised" are to be interpreted in the same manner.

[0036] "Essentially consisting of" means, in the present context, that the compound of interest included in a composition, such as one or more compounds of formula (1), is a major compound of the composition, i.e. the total amount of the compound is at least 70 wt% and up to about 95-98 wt.%, and that the composition further comprises about 2-30 wt % one or more other compounds that are typically structurally different from the compound of interest.

[0037] "Effective amount" means an amount of a composition that provides a synthetic complex lipid, or a salt thereof in a sufficient amount to render a desired treatment outcome in a young child. An effective amount can be administered in one or more doses to achieve the desired treatment outcome.

[0038] "Treat" or "treating" means in the present context to give a care or attention to a human subject, in context of the present invention, a young child, by providing said human subject with dietary means that supports a healthy body functioning, helps to relieve the symptoms of pathogen infections, allergy and / or inflammation in this human subject, or help to heal or cure an inflammatory condition or disease, or reduce the likelihood of developing thereof in said human subject by addressing nutritional need of said subject. "Treating" and "treatment" have grammatically corresponding meanings.

[0039] "Enteral administration" means any conventional form for delivery of a composition to a human subject that causes the deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include feeding through a naso-gastric tube or jejunum tube, oral, sublingual and rectal.

[0040] "Oral administration" means any conventional form for the delivery of a composition to a human subject, such as a young child, through the mouth. Accordingly, oral administration is a form of enteral administration. Oral administrations is a preferred method of administration of compositions of the invention.

[0041] "Milk fat globule membrane" means the 3-layered membrane surrounding the lipid droplets in milk and which maintains the lipid dispersion in the aqueous phase of milk. Milk fat globule membrane is highly structured and contains various components, such as glycoproteins, polar lipids in the form of phospholipids and glycosphingolipids, and apolar lipids in the form of cholesterol and cerebrosides. It is usually abbreviated as MFGM.

[0042] "Probiotic" means a live microorganism which when administered in adequate amounts confers a health benefit on the host.

[0043] The term "a" grammatically is a singular, but it may as well mean the plural of e.g., the intended compound. For example, a skilled person would understand that in the expression "a glycosphingolipid", the provision of not only one single glycosphingolipid, but of a variety of glycosphingolipids of the same type is meant.

[0044] As used herein, the term "alkyl" refers to an acyclic straight or branched hydrocarbyl group having 1- 50 carbon atoms which may be saturated or contain one or more double and / or triple bonds (so, forming for example an alkenyl or an alkynyl), and / or which may be substituted or unsubstituted, as herein further described. Examples of "alkyl" include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, sec-butyl, tert-butyl, isopentyl, n-pentyl, neo-pentyl, n-hexyl, ethenyl, propenyl, 1-butenyl, 2-butenyl, isobutenyl,l-pentenyl, 2-pentenyl, 2-methyl-l-butenyl, 3-methyl-l- butenyl, 2-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, methylpentenyl, dimethylbutenyl, ethynyl, propynyl, 1-butynyl, 2-butynyl, pentynyl, and hexynyl, each of which may be substituted or unsubstituted. Typically, the term alkyl refers to a straight saturated acyclic hydrocarbyl group having 13-19 carbons, which may be substituted or unsubstituted, preferably unsubstituted.

[0045] The skilled person would understand that when speaking of position C-l, C-2, C-3, C-4, C-5 etc., reference is herein always made to the respective carbon atoms of glycosphingolipid such as those represented by formula (l)-(6).

[0046] The skilled person will understand that in formulas showing a specific compound, like for example formulas (l)-(6) unless the chemical formula expressly describes a carbon atom having a particular stereochemical configuration, the formula is intended to cover compounds where such a stereocenter has an R or an S configuration, or wherein a double bond has a cis or a trans configuration.

[0047] The term "glycosyl moiety" refers to a monosaccharide or an oligosaccharide (more than one monosaccharide units), wherein the anomeric carbon of the monosaccharide or the anomeric carbon at the reducing end of the oligosaccharide is engaged in a glycosidic bond with another chemical entity, such as a sphingolipid, and the bond, if not further specified, may be an alpha or a beta glycosidic bond. A glycosyl moiety having more than one monosaccharide unit may represent a linear or a branched structure.

[0048] The monosaccharide unit is preferably any 5-9 carbon atom sugar, comprising aldoses (e.g. D- glucose, D-galactose, D-mannose, D-ribose, D-arabinose, L-arabinose, D-xylose, etc.), ketoses (e.g. D- fructose, D-sorbose, D-tagatose, etc.), deoxysugars (e.g. L-rhamnose, L-fucose, etc.), deoxyaminosugars (e.g. N-acetylglucosamine, N-acetylmannosamine, N-acetylgalactosamine, etc.), uronic acids, ketoaldonic acids (e.g. sialic acid). The monosaccharide unit can form different cyclic structures such as pyranose (six-membered) cyclic structures or furanose (five-membered) cyclic structures. In some embodiments the glycosyl moiety derives from a monosaccharide, wherein the monosaccharide is a p-galactoside. In some embodiments, the glycosyl moiety derives from an oligosaccharide, wherein the oligosaccharide carries one or more terminal p-galactopyranosyl units.

[0049] The glycosyl moieties according to the present invention may be illustrated in the following style: Gaipi-4Glcl-, wherein the dash (-) represents the point of attachment of the glycosyl moiety and wherein the glycosyl moiety, may be linked via an alpha or a beta glycosidic bond, preferably a beta glycosidic bond.

[0050] In the context of the present invention, the term Glc is glucosyl moiety, the term Gal is a galactosyl moiety, and term Neu5Ac is a / V-acetylneuraminosyl moiety.

[0051] In one aspect, the present invention provides a synthetic composition comprising one or more compounds of formula (1), or salts thereof: wherein

[0052] R1is a C13-C17 alkyl chain, preferably a C13 alkyl chain ;

[0053] R2is a C15-C19 alkyl chain, preferably, C15 or C17 alkyl chain; and

[0054] R3is a carbohydrate moiety selected from Neu5Aca2-3-Gaipi-4-Glcpi-, or Neu5Aca2-8-Neu5Aca2-3- Gaipi-4-Glc pi-.

[0055] In one aspect, the present invention provides a composition essentially consisting of one or more compounds of formula (1), or salts thereof: wherein

[0056] R1is a C13-C17 alkyl chain, preferably a C13 alkyl chain ;

[0057] R2is a C15-C19 alkyl chain, preferably, C15 or C17 alkyl chain; and

[0058] R3is a carbohydrate moiety selected from Neu5Aca2-3-Gaipi-4-Glcpi-, or Neu5Aca2-8-Neu5Aca2-3- Gaipi-4-Glc pi-.

[0059] In the context of the present invention, compounds of formula (1) may also be referred to as "glycosphingolipid", wherein the term "glycosphingolipid" refers to a compound that structurally consists of a glycosyl moiety and a sphingolipid moiety. The glycosyl moiety is typically linked to the sphingolipid moiety via a glycosidic bond between the anomeric carbon at the reducing end of the glycosyl moiety and the hydroxyl group at the C-l position of the sphingolipid.

[0060] According to the invention compounds of formula (1) are synthetic compounds, i.e. compounds produced by man via a synthetic process.

[0061] The term "synthetic composition" as used herein refers to a composition which is artificially prepared and preferably refers to a composition containing at least one compound that is produced ex vivo chemically and / or biologically, e.g. by means of chemical reaction, enzymatic reaction or recombinantly. Synthetic compositions of the invention either essentially consist of one or more synthetic glycosphingolipids described herein or comprising thereof, e.g., in some embodiments, the synthetic compositions may comprise one or more nutritionally or pharmaceutically active components (natural or synthetic) which do not adversely affect the efficacy of synthetic glycosphingolipids of the invention, but rather supplementary to the synthetic glycolipids of the invention in their biological action. All compositions according to the invention described herein are synthetic compositions. Accordingly, the terms "composition" and "synthetic composition" are used in the context of the invention as synonyms and relate to synthetic compositions, unless specified otherwise. The term "compound of formula" in the context of the invention relates to synthetic compounds, unless specified otherwise. The sphingolipid moiety of the glycosphingolipid of the present invention typically derives from a ceramide.

[0062] Ceramides denote in the context of the present invention naturally occurring ceramides, analogues thereof or derivatives thereof. Preferred ceramides are those naturally occurring in humans. Naturally occurring human ceramides [Cer] include, but are not limited to, Cer[NS], Cer[AS], Cer[EOS], Cer[NH], Cer[AH], or Cer[EOH], Cer[NP], Cer[AP], or Cer[EOP], Cer[NDS], Cer[ADS], or Cer[EODS], The letters in brackets refer to the shorthand nomenclature developed by Motta et al. (1993) Biochim Biophys Acta. 1182:147-151 and expanded by Rabionet (2014) Biochim Biophys Acta. 1841:422-434 and Masukawa et al., Journal of Lipid Research, 2008, 49, 1466-1476. Particularly, the letters N, A, and EO represent non-hydroxy fatty acids (N), alpha-hydroxy fatty acids (A), and omega-linoleoyloxy fatty acids (EO), respectively, wherein the number of fatty acid carbons and unsaturations may be expressed in parentheses following the letters of N, A, E, and O. The letters, S, H, P, and DS represent D-erythro-sphingosine (S), 6-hydroxy-D-erythro-sphingosine (H), D-ribo- phytosphingosine (P), D-erythro-dihydrosphingosine (DS), respectively, wherein the number of sphingoid carbons may be expressed in parenthesis following the letters S, H, P, and DS. Ceramides, Cer[NDS], Cer[ADS], or Cer[EODS], may also be referred to as Cer[NG], Cer[AG], or Cer[EOG], respectively, wherein the letter G represents the INCI name for D-erythro-dihydrosphingosine.

[0063] Glycosphingolipids lacking the amide-linked fatty acyl group may also be referred to as lysosphingolipids.

[0064] Both ceramides and lysosphingolipids used for the purposes of the invention are preferably of synthetic origin.

[0065] In some embodiments, said composition comprising one compound of formula (1).

[0066] In some embodiments, said composition essentially consists of one compound of formula (1).

[0067] In some embodiments, the compound of formula (1) is a compound of formula (2), or a salt thereof: wherein, R1and R2are defined as for the compound of formula (1).

[0068] In some preferred embodiments, the compound of formula (1) is a compound of formula (2), wherein the compound of formula (2) is the ganglioside GM3.

[0069] In some preferred embodiments, the present invention provides a composition comprising the ganglioside GM3. In some other preferred embodiments, the present invention provides a composition comprising a synthetic human identical GM3 (i.e. structurally identical molecule to a human ganglioside GM3). By the term "human ganglioside GM3" in the present context is meant a molecule of formula (1) wherein R1is preferably a C13-C17 alkyl chain; R2is preferably a C15-C19 alkyl chain; and R3is the carbohydrate moiety Neu5Aca2-3-Gaipi-4-Glcpi-., i.e. a compound of formula (2).

[0070] In some embodiments, the compound of formula (1) is a compound of formula (3), or a salt thereof Galpl^-Glcpl-O^ ^^-R1Neu5Aca2-3 J.

[0071] | OH

[0072] Neu5Aca2-8

[0073] (3), wherein R1and R2are as defined as for the glycosphingolipid of formula (1).

[0074] In some preferred embodiments, the compound of formula (1) is a compound of formula (3), wherein the compound of formula (3) is the ganglioside GD3.

[0075] In some preferred embodiments, the present invention provides a composition comprising the ganglioside GM3.

[0076] In other preferred embodiments, the composition may essentially consist of GM3.

[0077] In some preferred embodiments, the ganglioside GD3 is a synthetic human identical ganglioside GD3 (i.e. a synthetic molecule that is identical to a human ganglioside GD3). By the term "human ganglioside GD3" is meant a molecule of formula (1), wherein R1is preferably a C13-C17 alkyl chain; R2is preferably a C15-C19 alkyl chain; and R3is the glycosyl moiety Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-, i.e. a compound of formula (3). In some preferred embodiments the present invention provides a synthetic composition comprising a human identical ganglioside GD3.

[0078] In some preferred embodiments the present invention provides a composition comprising the ganglioside GD3.

[0079] In some preferred embodiments the present invention provides a composition essentially consisting of ganglioside GD3.

[0080] In some embodiments, the present invention provides a composition essentially consisting of more than one compound of formula (1). In some embodiments, the present invention provides a composition comprising more than one compound of formula (1).

[0081] In some preferred embodiments, the composition comprising a mixture of a compounds of formula (2) and a compound of formula (3), or salts thereof: wherein R1and R2are as defined as for the glycosphingolipid of formula (1).

[0082] In some other preferred embodiments, the composition essentially consists of a mixture of a compound of formula (2) and a compound of formula (3), or salts thereof:

[0083] (3), wherein R1and R2are as defined as for the glycosphingolipid of formula (1).

[0084] In other preferred embodiments, the composition comprising a compound of formula (2) and a compound of formula (3).

[0085] In preferred embodiments, the composition comprising a compound of formula (2) and a compound of formula (3), wherein the compound of formula (2) is a human identical synthetic ganglioside GM3 and the compound of formula (3) is a human identical synthetic ganglioside GD3. Accordingly, in some preferred embodiments, the present invention provides a composition consisting of the human identical gangliosides GM3 and GD3 .The term "human identical ganglioside" in the present context is meant a ganglioside molecule of any of formula (1), wherein R1is a C13-C17 alkyl chain; R2is preferably a C15-C19 alkyl chain; and R3is the carbohydrate moiety Neu5Aca2-3-Gaipi-4-Glcpi- or Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-., preferably, R1is a C15-C17 alkyl chain; R2is preferably a C15-C17 alkyl chain.

[0086] In some embodiments, the composition comprising of one or more compounds if formula (1) may further comprise a compound of formula (4): wherein R1and R2are defined as for the compound of formula (1).

[0087] In some embodiments, the compound of formula (4) is lactosylceramide [N(18) S(18)]. Lactosylceramide [N(18) S(18)] may also be referred to LacCer[N(18) S(18)].

[0088] In some embodiments, the composition essentially consisting of one or more compounds of formula (1) may further comprise a compound of formula (4): wherein R1and R2are defined as for the compound of formula (1). In some embodiments, the composition may further comprise a compound of formula (5):

[0089] (5), wherein R1are R2are defined as for the compound of formula (1).

[0090] In some embodiments, the compound of formula (5) is glucosylceramide [N(18) S(18)].

[0091] Glucosylceramide [N(18) S(18)] may also be referred to as GlcCer[N(18) S(18)].

[0092] In some embodiments, the composition may further comprise a compound of formula (6), or a salt thereof:

[0093] (6), wherein R1and R2are defined as for the compound of formula (1).

[0094] In some embodiments, the compound of formula (6) is the ganglioside GT3.

[0095] In some embodiments, the compound of formula (6) isa human-identical ganglioside GT3.

[0096] In some preferred embodiments, R1of the compounds of formula (l)-(6) is an unsubstituted C13 alkyl chain.

[0097] In some embodiments, R2of the compounds of formula (l)-(6) is an unsubstituted C15-C19 alkyl chain.

[0098] In one embodiment, R2of the compounds of formula (l)-(6) is an unsubstituted C15 alkyl chain.

[0099] In one preferred embodiment, R2of the compounds of formula (l)-(6) is an unsubstituted C17 alkyl chain.

[0100] In one embodiment, R2of the compounds of formula (l)-(6) is an unsubstituted C19 alkyl chain.

[0101] In some embodiments, R2of the compounds of formula (l)-(6) is a substituted C15-C19 alkyl chain. In some embodiments, the composition of the present invention comprising more than one compound of formula (l)-(6) which differ from each other in the number of carbon atoms in the alkyl chain of R2.

[0102] In one preferred embodiment, said composition comprising a compound of formula (1) wherein R2is a C15 alkyl chain and a compound of formula (1) wherein R2is a Ci7alkyl chain. Preferably the mass ratio of the compound of formula (1) wherein R2is a Ci5alkyl chain and the compound of formula (1) wherein R2is a Ci7alkyl chain is respectively from about 1:1 to about 1:4, for example from about 1:1.5 to about 1:3. In one embodiment, the mass ratio of the compound of formula (1) wherein R2is a C15 alkyl chain and the compound of formula (1) wherein R2is a Ci7alkyl chain is respectively between about 1:1.5 to about 1:2.5.

[0103] In one embodiment, said composition comprising a compound of formula (1) wherein R2is a Ci5alkyl chain, a compound of formula (1) wherein R2is a Ci7alkyl chain and a compound of formula (1) wherein R2is a Cig alkyl chain, and wherein the ratio between the compound of formula (1) wherein R2is a Cis alkyl chain, the compound of formula (1) wherein R2is a Ci7alkyl chain, and the compounds of formula (1) wherein R2is a Cig alkyl is respectively from about 1:1:1 to about l:4:0.1, for example about 1:1.5:1 to about l:3:0.1. If said composition comprises more than one compound of formula (1), Ri is preferably an unsubstituted C13 alkyl chain.

[0104] In some embodiments, the composition of the present invention comprising the ganglioside GM3.

[0105] In some embodiments, the composition of the present invention is essentially consisting of the ganglioside GM3.

[0106] In some embodiments, the composition of the present invention comprising at least about 70 wt.% of GM3, or at least about 75 wt.% of GM3, or at least about 80 wt.%> of GM3, or at least about 85 wt. i of GM3.

[0107] In some embodiments, the composition of the present invention comprising the ganglioside GM3, and wherein said composition further comprising LacCer[N(18) S(18)] and GlcCer[N(18) S(18)], respectively.

[0108] In some embodiments, the composition of the present invention comprising about 70-85 wt.%> of GM3, and wherein said composition further comprising about 5-10 wt.%> or LacCer[N(18) S(18)] and about 0.5-1 wt. of GlcCer[N(18) S(18)].

[0109] In some embodiments, the composition of the present invention comprising the ganglioside GM3 and the ganglioside GD3 respectively. In some embodiments, the composition of the present invention is essentially consisting of the ganglioside GM3 and the ganglioside GD3.

[0110] In some embodiments, the composition of the present invention comprising the ganglioside GM3 and the ganglioside GD3, and wherein the mass ratio between GM3 and GD3 in said mixture is from about 0.2:14 to 14:0.2. For example, in some embodiments, the mass ratio between GM3 and GD3 in said composition may be about 14:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 0.2:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 5:1.

[0111] In some embodiments, the composition of the present invention essentially consisting of the ganglioside GM3 and the ganglioside GD3, and wherein the mass ratio between GM3 and GD3 in said mixture is from about 0.2:14 to 14:0.2. For example, in some embodiments, the mass ratio between GM3 and GD3 in said composition may be about 14:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 0.2:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 5:1.

[0112] In some embodiments, the composition of the present invention comprising the ganglioside GM3 and the ganglioside GD3, and wherein the mass ratio between GM3 and GD3 in said mixture is from about 1:10 to 10:1. For example, in some embodiments, the mass ratio between GM3 and GD3 in said composition may be about 10:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 3:1. In some other embodiments, the mass ratio between GM3 and GD3 in said composition may be about 1:1. In some embodiments, the mass ratio between GM3 and GD3 in said composition may be about 1:4.

[0113] In some preferred embodiments, the composition essentially consists of the ganglioside GM3 and the ganglioside GD3, and wherein the mass ratio between GM3 and GD3 in said mixture is from about 1:10 to 10:1. For example, in some embodiments, the mass ratio between GM3 and GD3 in said composition may be about 10:1. In some other preferred embodiments, the mass ratio between GM3 and GD3 in said composition may be about 3:1. In some other preferred embodiments, the mass ratio between GM3 and GD3 in said composition may be about 1:1. In some preferred embodiments, the mass ratio between GM3 and GD3 in said composition may be about 1:4.

[0114] In some embodiments, the composition of the present invention comprising the ganglioside GM3 and the ganglioside GD3, and wherein said composition further comprising LacCer[N(18) S(18)] and GlcCer[N(18) S(18)].

[0115] In some embodiments, the composition essentially consisting of the ganglioside GM3 and the ganglioside GD3 further comprises LacCer[N(18) S(18)] and GlcCer[N(18) S(18)]. In some embodiments, the composition of the present invention may comprise the ganglioside GM3 and the ganglioside GD3, and wherein said composition further comprising LacCer[N(18) S(18)], GlcCer[N(18) S(18)] and the ganglioside GT3.

[0116] In some embodiments, the composition of the present invention comprises about 60-65 wt.% of the ganglioside GM3, about 20-25 wt.% of the ganglioside GD3, and wherein said composition further comprising about 5-10 wt.% LacCer[N(18) S(18)] and about 0.4-1 wt.% of GlcCer[N(18) S(18)].

[0117] In some embodiments, the composition of the present invention may comprise about 10-15 wt.% of the ganglioside GM3, about 40-55 wt.% of the ganglioside GD3, and wherein said composition further comprising about 4-6 wt.% of the ganglioside GT3, about 5-7 wt.% LacCer[N(18) S(18)] and about 0.4-1 wt.% of GlcCer[N(18) S(18)].

[0118] In some embodiments, the composition of the present invention may comprise up to about 95-98 wt.% gangliosides described therein and about 2-5 wt.% of LacCer[N(18) S(18)] and GlcCer[N(18) S(18)] together.

[0119] The salt of the compound of formulas (1), (2), (3), and (6) can be any suitable salt which is compatible with food use. For example, a sodium, potassium, lithium, calcium, magnesium, zinc, aluminium, trietylamine, diethanolamine, ethanolamine, ethylenediamine, arginine, lysine, histidine, choline, benzathine, chloroprocaine, procaine, or meglumine salt.

[0120] In one embodiment, the composition of the present invention can be obtained by a method comprising the steps of :

[0121] - Reacting 3'-sialyllactose, an enzyme having trans-sialidase activity, an enzyme having p- galactosidase activity, and a compound formula (7), or a salt thereof:

[0122] NH2Galpl-4-Glcpl-O^ ^^

[0123] OH

[0124] (7), wherein R1is as defined as for the compound of formula (1); to obtain a composition comprising a compound of formula (8), or a salt thereof: (paipi-4-Glcpi- Neu5Aca2,3

[0125] (8), wherein R1is as defined as for the compound of formula (1);

[0126] - Reacting the composition comprising the compound of formula (8) with one or more triazine triazine-based acylating agents of formula (9):

[0127] Oh wherein i i represents a conjugated system of bonds such that either two or three double bonds are present in the ring,

[0128] Y is selected from C(O-C(=O)R2), or C(=O),

[0129] Xais selected from N, NR4, or N(C(=O)R2)

[0130] Xbis selected from N, or NR4

[0131] Z is selected from C(=O), or C(OR4), and provided that: when Y is C(O-C(=O)R2), Z is C(OR4), Xaand Xbare N, and three double bonds are present in the ring, or when Y is C(O-C(=O)R2), Z is C(=O), one of Xaand Xbis N and the other group is NR4, and two double bonds are present in the ring, or when Y is C(=O), Z is C(OR4), Xais N(C(=O)R2), Xbis N, and two double bonds are present in the ring, and wherein

[0132] R2is as defined as for the compound of formula (1),

[0133] R4is selected from methyl, ethyl, 2,2,2-trifluoroethyl, and substituted or unsubstituted benzyl; to produce the composition comprising the compound of formula (2).

[0134] In another embodiment, the composition of the present invention can be obtained by a method comprising the steps of: - Reacting 3'-sialyllactose, an enzyme having trans-sialidase activity, an enzyme having p- galactosidase activity, and a compound formula (7), or a salt thereof:

[0135] Gaip 1 -4-Glcp wherein R1is as defined as for the compound of formula (1); to obtain a composition comprising a compound of formula (8), or a salt thereof:

[0136] NH2

[0137] (jSaipi -4-Glc i-O^ ^^- / R1

[0138] Neu5Aca2,3 OH

[0139] (8), wherein R1is as defined as for the compound of formula (1);

[0140] - Reacting the composition comprising the compound of formula (8) with a mixture comprising sialic acid, cytidine monophosphate (CMP), a nucleoside triphosphate, and at least five enzymes, in which the at least five enzymes comprise at least one enzyme having sialyltransferase activity, at least one enzyme having a / V-acylneuraminate citydyltransferase activity, at least one enzyme having inorganic diphosphatase activity, and at least two enzymes having kinase activity; to obtain a composition comprising a compound of formula (10), or a salt thereof:

[0141] Gaipi-4-Glcpl- Neu5Aca2-3

[0142] |

[0143] Neu5Aca2-8

[0144] (10), wherein R1is as defined as for the compound of formula (1);

[0145] - Reacting the composition comprising the compound of formula (10) with one or more triazine triazine-based acylating agents of formula (9): represents a conjugated system of bonds such that either two or three double bonds are present in the ring,

[0146] Y is selected from C(O-C(=O)R2), or C(=O), Xais selected from N, NR4, or N(C(=O)R2) Xbis selected from N, or NR4Z is selected from C(=O), or C(OR4), and provided that: when Y is C(O-C(=O)R2), Z is C(OR4), Xaand Xbare N, and three double bonds are present in the ring, or when Y is C(O-C(=O)R2), Z is C(=O), one of Xaand Xbis N and the other group is NR4, and two double bonds are present in the ring, or when Y is C(=O), Z is C(OR4), Xais N(C(=O)R2), Xbis N, and two double bonds are present in the ring, and wherein

[0147] R2is as defined as for the compound of formula (1),

[0148] R4is selected from methyl, ethyl, 2,2,2-trifluoroethyl, and substituted or unsubstituted benzyl; to produce the composition comprising the compound of formula (3), or the composition comprising the compounds of formula (2) and of formula (3).

[0149] "Enzyme having a p-galactosidase activity" means an enzyme belonging to the glycoside hydrolase family 35 (GH35) which typically catalyses the hydrolysis of terminal non-reducing p-d-galactose residues in p-d-galactosides. An enzyme having a p-galactosidase activity is used interchangeably with the term "P-galactosidase". A p-galactosidase may also be referred to as lactase.

[0150] "Enzyme having a trans-sialidase activity" means an enzyme belonging to the glycoside hydrolase family 33 (GH33) which typically catalyzes the reversible transfer of a glycosidically linked sialic acid from sialic acid donors such as oligosaccharides, glycoproteins, glycolipids, and colominic acid to acceptor molecules containing a terminal p-galactopyranosyl unit. Enzyme having a trans-sialidase activity is used interchangeably with the term "trans-sialidase". In the absence of a suitable acceptor molecule, an enzyme having a trans-sialidase activity may act as a sialidase and transfer a glycosidically linked sialic acid to a water molecule. However, the hydrolytic activity is typically low.

[0151] "Enzyme having inorganic diphosphatase activity" means an enzyme that catalyzes the hydrolysis of pyrophosphate (PPi). An enzyme having inorganic diphosphatase activity is used interchangeably with the term "inorganic diphosphatase" or "PPase".

[0152] "Enzyme having / V-acylneuraminate cytidyltransferase activity" means an enzyme that catalyzes the transfer of CMP from CTP to / V-acetyl-neuraminic acid (Neu5Ac). An enzyme having N- acylneuraminate cytidyltransferase activity is used interchangeably with the term "N- acylneuraminate cytidylyltransferase" or "CSS".

[0153] "Enzyme having a nucleoside diphosphate kinase activity" means an enzyme that catalyzes the phosphorylation of a nucleoside diphosphate. An enzyme having a nucleoside diphosphate kinase activity is used interchangeably with the term "nucleoside-diphosphate kinase" or "NDK".

[0154] "Enzyme having a sialyltransferase activity" means an enzyme belonging to the glycosyltransferase family 29 (GT29), or to the glycosyltransferase family 42 (GT42) which typically catalyzes the transfer of sialic acid from CMP-sialic acid to a saccharide acceptor. Enzyme having a sialyltransferase activity is used interchangeably with the term "sialyltransferase" and

[0155] The enzyme having trans-sialidase activity in its wild-type form, may originate from parasitic euglenoids, such as Trypanosoma cruzi, Trypanosoma congolense, or Trypanosome brucei.

[0156] In some embodiments, the enzyme having trans-sialidase activity is a wild-type trans-sialidase originating from Trypanosoma cruzi. The amino acid sequence of the wild-type trans-sialidase originating from Trypanosoma cruzi can be found on https: / / www.uniprot.org / , accession: Q26966. The trans-sialidase originating from Trypanosoma cruzi may also be referred to as TcTS.

[0157] In some embodiments, the enzyme having trans-sialidase activity is a mutant of the wild-type trans- sialidase originating from Trypanosoma cruzi (Q26966). In some embodiments, the mutant trans- sialidase has at least five mutations at amino acid positions selected from the following positions (numbered corresponding to alignment of the amino acid sequence with the amino acid sequence of Q26966): S263T, R477H, V485L, E559V, N59F, S496K, V497G, E521K, D594G, I598D and H600R. In some embodiments, the amino acids sequence of the mutant trans-sialidase is that described on https: / / www.rcsb.org / structure / lMSO. The mutant trans-sialidase may be produced by methods known to the skilled person. A method for the expression and purification of a mutant trans-sialidase is for example described in Paris et al., Glycobiology 2001, 11, 305-311, or in Buschiazzo et al., Molecular Cell 2002, 10, 757-768.

[0158] The enzyme having p-galactosidase activity in its wild-type form, may originate from Aspergillus oryzae. In some preferred embodiments, the enzyme having p-galactosidase activity is a truncated variant of the wild-type p-galactosidase originating from Aspergillus orizyae (Q2UCU3). The truncated variant of the p-galactosidase can be purchased from established manufacturers, e.g. Calza Clemente, or produced by methods known to the skilled person such as that described in M.M. Maksimainen et al., International Journal of Biological Macromolecules 2013, 60, 109-115.

[0159] In some embodiments, the enzyme having sialyltransferase activity is a wild-type a-2,3 / a-2,8- sialyltransferase originating from Campylobacter jejuni, strain OX=197, or a functional analogue thereof. The amino acid sequence of the wild-type a-2,3 / a-2,8-sialyltransferase originating from Campylobacter jejuni, strain OX=197 can be found on https: / / www.uniprot.org / , accession: Q9LAK3. The a-2,3 / a-2,8-sialyltransferase originating from Campylobacter jejuni is referred to as CST-II.

[0160] In some embodiments, the enzyme having sialyltransferase activity is a mutant of the wild-type a- 2,3 / a-2,8-sialyltransferase originating from Campylobacter jejuni, strain OX=197 (Q9LAK3). In some embodiments, the mutant a-2,3 / a-2,8-sialyltransferase has a mutation at the amino acid 15 position I53G (numbering corresponding to alignment of the amino acid sequence with the amino acid sequence of Q9LAK3), as described in Gilbert et al., Biological Chemistry 2002, 277, 327-337.

[0161] In some embodiments, the triazine-based acylating agents of formula (9) is a triazine-based acylating agent selected from triazine based acylating agents of formulas (11) and (12): wherein R2is as defined as the compound of formula (1), and R4is as defined as for the triazine-base acylating agent of formula (9). In some embodiments, triazine triazine-based acylating agents of formula (9) is a triazine-based acylating agent selected from triazine based acylating agents of formulas (13) and (14): wherein R2is as defined as the compound of formula (1), and R4is as defined as for the triazine-base acylating agent of formula (9).

[0162] The compound of formula (7) and the triazine base-acylating agents of formulas (9) and (11)-(14) can be obtained by method known to the skilled person. For instance, a method to produce a compound of formula (7) is described by Vaughan et al., J. Am. Chem. Soc. 2006, 128, 6300-6301. A method to produce a triazine-based acylating of formula (9) and (11)-(14) is for example described by Kaminski et al., Journal , prakt. Chemie. 1990, Band 332, Heft 4, S. 579-383. A detailed description of the method of producing the composition according to the present invention can be found in the examples below.

[0163] In some preferred embodiments, the nucleoside triphosphate is adenosine 5'-triphosphate (ATP), and the at least five enzymes comprise the following enzyme:

[0164] - an enzyme having sialyltransferase activity,

[0165] - an enzyme having cytidine monophosphate kinase activity,

[0166] - an enzyme having nucleoside diphosphate kinase activity,

[0167] - an enzyme having N-acylneuraminate cytidyltransferase activity, and

[0168] - an enzyme having inorganic diphosphatase activity.

[0169] In some preferred embodiments, the enzyme having cytidine monophosphate kinase activity is a wild-type CMP kinase originating from Mycobacterium tuberculosis, or a functional analogue thereof. The amino acid sequence of the wild-type CMP kinase originating from Mycobacterium tuberculosis can be found on https: / / www.ncbi.nlm.nih.gov / genbank / , accession: WP_129368399. The CMP kinase originating from Mycobacterium tuberculosis is also referred to as MtCMK. In some embodiments, the enzyme having nucleoside diphosphate kinase activity is a wild-type nucleoside-diphosphate kinase originating from Mycobacterium tuberculosis complex, or a functional analogue thereof. The amino acid sequence of the wild-type nucleoside-diphosphate kinase originating from Mycobacterium tuberculosis complex, can be found on https: / / www.ncbi.nlm.nih.gov / genbank / , accession: WP_003412592. The nucleoside-diphosphate kinase originating from Mycobacterium tuberculosis complex may also be referred to as MtNDK.

[0170] In some preferred embodiments, the enzyme having / V-acylneuraminate cytidyltransferase activity is a wild-type / V-acylneuraminate cytidylyltransferase originating from Neisseria meningitidis or a functional analogue thereof. The amino acid sequence of the wild-type / V-acylneuraminate cytidylyltransferase originating from Neisseria meningitidis can be found on https: / / www.ncbi.nlm.nih.gov / genbank / , accession: WP_061726245. The / V-acylneuraminate cytidyltransferase originating from Neisseria meningitidis is also referred to as NmCSS.

[0171] In some embodiments, the enzyme having inorganic diphosphatase activity is a wild-type inorganic diphosphatase originating from Escherichia coli, or a functional analogue thereof. The amino acid sequence of the wild-type inorganic diphosphatase originating from Escherichia coli can 30 be found on https: / / www.ncbi.nlm.nih.gov / genbank / , accession: WP_073849715. The inorganic diphosphatase originating from Escherichia coli is also referred to as EcPPase.

[0172] The mutated variant and the wild-type enzymes used during the sialyltransferase cycle, can be purchased from established manufacturers, or produced by methods known to the skilled person. For instance, the mutant of the wild-type a-2,3 / a-2,8-sialyltransferase originating from Campylobacter jejuni can be produced by the method described in Gilbert et al., Biological Chemistry 152002, 277, 327-337, whereas the wild-type enzymes can be produced by the method described elsewhere in molecular cloning textbooks.

[0173] In another aspect, the invention provides a child nutritional formula which comprises the composition as defined above. The composition, as defined above comprising one or more compounds of formula (1), and in some embodiment further comprising a compound of formula (4),

[0174] (5), and / or (6).

[0175] In another aspect, the invention provides a child nutritional formula which comprises a composition as any of the described above, the composition essentially consisting of one or more compounds of formula (1), and, in some embodiments, further comprising a compound of formula (4), (5), and / or

[0176] (6).

[0177] In some embodiments, the child nutritional formula comprising the composition according to the present invention, wherein said composition comprising gangliosides GM3 and GD3, wherein the total content of gangliosides GM3 and GD3 in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l, and wherein the ratio between gangliosides GM3 and GD3 is from about 0.2:1 to about 14:1, preferably from about 0.2:1 to 5:1.

[0178] In some embodiments, the child nutritional formula comprising the composition according to the present invention, wherein said composition comprising ganglioside GM3, and wherein the total content of GM3 in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

[0179] In some embodiments, the child nutritional formula comprising the composition according to the present invention, wherein said composition comprising gangliosides GM3 and GD3, and wherein said composition further comprising LacCer[N(18) S(18)], GlcCer[N(18) S(18)] and ganglioside GT3.

[0180] In some embodiments, the child nutritional formula comprising the composition according to the present invention, wherein said composition comprising gangliosides GM3, and wherein said composition further comprising LacCer[N(18) S(18)] and GlcCer[N(18) S(18)].

[0181] In one embodiment, the child nutritional formula comprises from about 0.001 mg / kcal to about 0.04 mg / kcal of one or more compounds compound of formula (2). For example, the child nutritional formula may comprise from about 0.003 mg / kcal to about 0.025 mg / kcal of the one or more compounds of formula (2), or about 0.004 mg / kcal to about 0.017 mg / kcal of the one or more compounds of formula (2). When in a ready to feed form, the child nutritional formula can contain from about 1 mg / l to about 20 mg / l of the one or more compounds of formula (2), for example about 2 mg / l to about 15 mg / l, or about 3 mg to about 10 mg / l. Any value within the given ranges is contemplated.

[0182] In one embodiment, the child nutritional formula comprises from about 0.0001 mg / kcal to about 0.017 mg / kcal of the compound of formula (3). For example, the child nutritional formula may comprise from about 0.0003 mg / kcal to about 0.008 mg / kcal of the compound of formula (3), or from about 0.004 mg / kcal to about 0.003 mg / kcal of the compound of formula (3). When in a ready to feed form, the child nutritional formula can contain from about 0.1 mg / l to about 10 mg / l of the compound of formula (3), for example from about 0.25 mg / l to about 5 mg / l, from about 0.5 mg to about 2 mg / l, from about 1 mg / l to about 6 mg / l, from about 3 mg to about 8 mg / l, or from about 4 mg / l to about 10 mg / l . Any value within the given ranges is contemplated.

[0183] In one embodiment, the child nutritional formula may comprise ba mixture of compound of formula (2) and compound of formula (3). Any combination of the amounts of these compounds described above are contemplated. The details of preferred combinations of these compounds are also earlier in this specification.

[0184] In one embodiment, the child nutritional formula comprises about 0.0001 mg / kcal to about 0.025 mg / kcal of the compound of formula (4). For example, the child nutritional formula may comprise from about 0.0003 mg / kcal to about 0.017 mg / kcal of the compound of formula (4), or from about 0.004 mg / kcal to about 0.008 mg / kcal of the compound of formula (4). When in ready to feed form, the child nutritional formula can contain from about 0.1 mg / l to about 15 mg / l of the compound of formula (4), for example from about 0.2 mg / l to about 10 mg / l, or from about 0.3 mg to about 5 mg / l.

[0185] In one embodiment, the child nutritional formula comprises from about 0.0001 mg / kcal to about 0.025 mg / kcal of the compound of formula (5). For example, the child nutritional formula may comprise about 0.0003 mg / kcal to about 0.017 mg / kcal of the compound of formula (5), or about 0.004 mg / kcal to about 0.008 mg / kcal of the compound of formula (5). When in ready to feed form, the child nutritional formula can contain about 0.1 mg / l to about 15 mg / l of the compound of formula (5), for example from about 0.2 mg / l to about 10 mg / l, or from about 0.3 mg to about 5 mg / l.

[0186] The above described amounts of glycolipids of formulas (1) to (5) are effective amounts according to the invention.

[0187] The child nutritional formula as any of described above can comprise a source of protein. The source of protein can be in the form of intact protein, partially hydrolyzed protein, extensively hydrolyzed protein, or amino acids. The protein source can be any source of protein which is suitable for infants, for example cow's milk protein, goat's milk protein, rice protein, pea protein, soya protein, and the like. The milk protein may be in the form of whey protein, casein, or combinations of whey and casein. The protein source can be supplemented with free amino acids such that it provides enough of each indispensable and conditionally indispensable amino acid in human breast milk.

[0188] The protein content of the child nutritional formula is preferably in the range of about 1.8 g / 100 kcal to about 3.2 g / 100 kcal (about 0.43 g to about 0.76 g carbohydrate per 100 KJ). If the protein source is an intact milk protein, the protein content is preferably in the range of about 1.8 g / 100 kcal to about 2.5 g / 100 kcal (about 0.43 g to about 0.6 g carbohydrate per 100 KJ). If the protein source is or includes a plant protein (e.g., rice, soya and the like), the protein content is preferably in the range of about 2.25 g / 100 kcal to about 2.8 g / 100 kcal (about 0.54 g to about 0.67 g carbohydrate per 100 KJ). If the protein source is a hydrolyzed protein, the protein content is preferably in the range of about 1.8 g / 100 kcal to about 2.8 g / 100 kcal (about 0.43 g to about 0.67 g carbohydrate per 100 KJ). If the protein source is free amino acids, the protein content is preferably in the range of about 2.8 g / lOOkcal to 3.1 g / 100 kcal. For certain infants, such as those suffering gastrointestinal pathologies with severe malabsorption or premature infants, the protein content can be higher, for example up to 4.1 g / 100 kcal (0.97 g / 100 KJ).

[0189] The child nutritional formula as any of described above can also comprise a digestible carbohydrate. Examples of digestible carbohydrates include lactose, maltose, sucrose, glucose, glucose syrup or dried glucose syrup, maltodextrins, and starch. Mixtures of carbohydrates can also be used. If the child nutritional formula is intended for preterm infants, the carbohydrate source preferably contains less or no lactose. The digestible carbohydrate content of the child nutritional formula is preferably in the range of from about 9 g / 100 kcal to about 14 g / 100 kcal (from about 2.2 g to about 3.3 g carbohydrate per 100 KJ).

[0190] The child nutritional formula as any of the described above can further comprise a source of lipid. The lipid source may be any lipid which is suitable for use in infant nutrition. Preferred lipid sources include milk fat, sunflower oil, rapeseed oil, safflower oil, egg yolk lipid, olive oil, coconut oil, palm oil, palm kernel oil, soybean oil, fish oil, and microbial fermentation oil containing. These oils may be in the form of high oleic forms such as high oleic sunflower oil and high oleic safflower oil. The lipid source may also be in the form of fractions derived from these oils such as palm olein, medium chain triglycerides (MCT), and esters of fatty acids such as linoleic acid, palmitic acid, stearic acid, linolenic acid, oleic acid, lauric acid, capric acid, caprylic acid, caproic acid, and the like. The lipid source can also include structured lipids (i.e., lipids that are modified chemically or enzymatically to change their structure). Preferably, the structured lipids are sn2 structured lipids, for example comprising triglycerides having an elevated level of palmitic acid at the sn2 position of the triglyceride. The lipid source can also include oils containing high concentrations of long-chain, polyunsaturated fatty acids such as arachidonic acid (ARA), docosahexaenoic acid (DHA), and / or eicosapentaenoic acid such as fish oils or microbial oils. If the Child nutritional formula is intended for preterm infants, the lipid source preferably contains MCT. For example, the lipid source can contain up to 40% by weight of MCT, preferably about 15% to about 35% by weight of the lipid source.

[0191] The lipid content of the child nutritional formula is preferably in the range of from about 4.4 g / 100 kcal to about 6 g / 100 kcal (from about 1.1 g to about 1.4 g lipid per 100 KJ). If the child nutritional formula contains ARA, the amount of ARA is preferably in the range of from about 2mg to about 20 mg ARA per 100 kcal, for example about 5mg to about 15mg ARA per 100 kcal. If the child nutritional formula contains DHA, the amount of DRA is preferably in the range of about 2mg to about 20 mg DHA per 100 kcal, for example about 5mg to about 15 DHA per 100 kcal. For example, the child nutritional formula can contain about 10 mg ARA per 100 kcal and about 10 mg DHA per 100 kcal. The mass ratio of n-6 to n-3 fatty acids can be from about 5: 1 to about 15: 1; for example, about 8: 1 to about 10: 1.

[0192] When in nutritionally complete form, the child nutritional formula contains all vitamins and minerals understood to be essential in the daily diet and in nutritionally adequate amounts. Minimum requirements have been established for certain vitamins and minerals. Minerals which are normally required include sodium, potassium, chloride, calcium, phosphorous, magnesium, iron, zinc, copper, iodine, selenium, manganese, molybdenum, and fluoride. In general, the molar ratio of calcium to available phosphorus is about 1:1 to about 2:1. Vitamins which are normally required include vitamin A, vitamin D, thiamine (vitamin Bl), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), vitamin B6, biotin (vitamin B7), folate (vitamin B9), vitamin BI2, vitamin C, vitamin E, and vitamin K. The child nutritional formula may further contain one or more carotenoids.

[0193] The child nutritional formula can contain a source of other essential nutrients such as choline. Suitable sources of choline include milk fat, milk fat fractions, phospholipids, and choline salts. In general, the child nutritional formula can contain about 7 mg / 100 kcal to about 50 mg / 100 kcal. The child nutritional formula can also contain a source inositol. For example, the child nutritional formula may contain about 4 mg / 100 kcal to about 150 mg / 100 kcal.

[0194] The child nutritional formula can also comprise at least one probiotic. If the probiotic is capable of producing lactic acid, a probiotic which produce L(+) lactic acid is preferred. Examples of probiotics include yeasts, such as Saccharomyces; and bacteria, such as the genera Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus. Specific examples of suitable probiotics are: Saccharomyces cereviseae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbruckii subsp. lactis, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus rhamnosus ( Lactobacillus GG), Lactobacillus sake, Lactococcus lactis, Lactobacillus reuterii, Micrococcus varians, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus halophilus, Streptococcus faecalis, Streptococcus thermophilus, Staphylococcus carnosus and Staphylococcus xylosus.

[0195] The child nutritional formula can also comprise a source of non-digestible carbohydrates. Suitable non-digestible carbohydrates include human milk oligosaccharides, galacto-oligosaccharides, fructo- oligosaccharide, polydextrose, and mixtures of these oligosaccharides. Preferably the child nutritional formula contains a source of human milk oligosaccharides, for example 2'-fucosyllactose, 3-fucosyllactose, di-fucosyllactose, lacto-N-fucopentaose 1, lacto-N-tetraose, lacto-N-neotetraose, 3'- sialyllactose, 6'-sialyllactose, disialyllacto-N-tetraose, and mixtures of these oligosaccharides. The child nutritional formula may comprise from about 0.02 g / lOOkcal to about 3 g / lOOkcal of non- digestible carbohydrates, for example from about 0.02 g / lOOkcal to about 2.25 g / lOOkcal.

[0196] In one embodiment, the source of non-digestible carbohydrates includes an acidic human milk oligosaccharide such as 3'-sialyllactose, 6'-sialyllactose, and / or disialyllacto-N-tetraose. The acidic human milk oligosaccharide can be present in an amount from 0.015 g / lOOkcal to about 0.23 g / 100 kcal, for example from about 0.02 g / 100 kcal to about 0.15 g / 100 kcal. This corresponds to the amount from about 0.1 g / l to about 1.5 g / l, for example about 0.13 g / l to about 1 g / l, in ready to feed liquid.

[0197] In another embodiment, the source of non-digestible carbohydrates includes a fucosyl human milk oligosaccharide such as 2'-fucosyllactose, 3-fucosyllactose, di-fucosyllactose, lacto-N-fucopentaose 1. The acidic human milk oligosaccharide can be present in an amount from about 0.015 g / lOOkcal to about 1.5 g / 100 kcal, for example from about 0.02 g / 100 kcal to about 1.0 g / 100 kcal. This corresponds to the amount from about 0.1 g / l to about 10 g / l, for example from about 0.13 g / l to about 6.7 g / l, in ready to feed liquid.

[0198] In another embodiment, the source of non-digestible carbohydrates includes a core human milk oligosaccharide such as lacto-N-tetraose and / or lacto-N-neotetraose. The core human milk oligosaccharide can be present in an amount from 0.015 g / lOOkcal to about 0.3 g / 100 kcal, for example from about 0.02 g / 100 kcal to about 0.23 g / 100 kcal. This corresponds to the amount from about 0.1 g / l to about 2 g / l, for example from about 0.13 g / l to about 1.5 g / l, in ready to feed liquid.

[0199] The child nutritional formula can also comprise a source of animal milk fat globule membrane, especially cow milk fat globule membrane. The source of animal milk fat globule membrane can be selected from whole animal milk or a concentrated fraction of animal. Suitable concentrated fractions of animal milk include buttermilk, beta serum and fat-fractions obtained during the production of whey protein concentrates or whey protein isolates. Sources of milk fat globule membrane in the form of concentrated fractions of animal milk are commercially available.

[0200] The child nutritional formula can comprise other substances which may be beneficial to the infants and toddlers such as lactoferrin, nucleotides, nucleosides, sphingomyelin, and the like. If desired, the child nutritional formula can comprise emulsifiers and stabilizers such as soy lecithin, citric acid esters of mono-and di-glycerides, and the like. This is especially the case if the child nutritional formula is provided in liquid form.

[0201] The child nutritional formula can be prepared in any suitable manner. For example, a child nutritional formula can be prepared by blending the protein source, the carbohydrate source, and the lipid source in appropriate proportions. If used, emulsifiers may be included in the blend. The vitamins and minerals may be added at this stage but are usually added later to avoid thermal degradation. Any lipophilic vitamins, emulsifiers and the like may be dissolved into the lipid source prior to blending. The ganglioside composition of the invention can be included in the lipid source prior to blending. Water, preferably water which has been subjected to reverse osmosis, may then be mixed in to form a liquid mixture.

[0202] The liquid mixture then can 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, for example a plate heat exchanger.

[0203] The liquid mixture then can be cooled to about 60°C to about 85°C, for example by flash cooling. The liquid mixture then can 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 then can be further cooled to add any heat sensitive components, such as vitamins and minerals. The pH and solids content of the homogenized mixture is conveniently standardized at this point.

[0204] If it is desired to produce a powdered child nutritional formula, the homogenized mixture is transferred to a suitable drying apparatus such as a spray drier or freeze drier and converted to powder. The powder preferably has a moisture content of less than about 5% by weight.

[0205] If it is desired to produce a liquid child nutritional formula, the homogenized mixture is filled into suitable containers, preferably aseptically. However, the liquid child nutritional formula also can be retorted in the container. Suitable apparatus for carrying out filling of this nature is commercially available. The liquid child nutritional formula may be in the form of a ready to feed formula having a solids content of about 10 to about 14% by weight or may be in the form of a concentrate, usually having a solids content of about 20 to about 26% by weight.

[0206] According to the invention, inclusion of the compositions of the invention in a child nutritional formula has multiple benefits for young children since synthetic glycolipids of the compositions, such as gangliosides GM3 and GD3 described herein, are powerful enhancers of the production of shortchain fatty acids (SCFA) by microbiome bacteria of the young children gut, especially, when these children are administered effective amounts of the glycolipids regularly for a period for at least 14 days.

[0207] By the term "short-chain fatty acids" or "SCFAs" is meant fatty acids of two to six carbon atoms. Derived from intestinal microbial (microbiota) fermentation of indigestible foods, SCFAs in human gut are acetic, propionic and butyric acid.

[0208] The terms "microbiota", "microflora" and "microbiome" mean a community of living microorganisms that typically inhabits a bodily organ or part, particularly the gastro-intestinal organs of non-infant humans. The most dominant members of the gastrointestinal microbiota include microorganisms of the phyla of Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, Synergistetes, Verrucomicrobia, Fusobacteria, and Euryarchaeota; at genus level Bacteroides, Faecalibacterium, Bifidobacterium, Roseburia, Alistipes, Collinsella, 8 / autia Coprococcus, Ruminococcus, Eubacterium and Dorea; at species level Bacteroides uniformis, Alistipes putredinis, Parabacteroides merdae, Ruminococcus bromii, Dorea longicatena , Bacteroides caccae, Bacteroides thetaiotaomicron, Eubacterium ha / lii, Ruminococcus torgues, Faecalibacterium prausnitzii, Ruminococcus lactaris, Collinsella aerofaciens, Dorea formicigenerans, Bacteroides vulgatus and Roseburia intestinalis. The gastrointestinal microbiota includes the mucosa-associated microbiota, which is located in or attached to the mucus layer covering the epithelium of the gastrointestinal tract, and luminal- associated microbiota, which is found in the lumen of the gastrointestinal tract.

[0209] In accordance with this invention, it has been discovered that a synthetic glycolipid of the invention, or a salt thereof, or a composition essentially consisting thereof, stimulates the production of SCFA by the young child (i.e. the host) GIT microbiome and increases the total amount SCFA, in particular the amount of acetate and butyrate, in the GIT by at least 5% after around 14 days of ingestion said glycolipid, salt or composition, and decreases the amount of produced by the microbiome propionate and branched-SCFA (b-SCFA). For this reason, the synthetic glycolipid of the invention, salt thereof or composition comprising thereof can be used as the acetate and butyrate- providing dietary supplement for dietary treatment of young children that would have beneficial effect on healthy growth and development of these young children by restoring, maintaining or improving intestinal tissue structure and mucus production by the intestinal goblet cells, the mucus quality, mucosal immune and inflammatory status and gut permeability, and reducing the risk of development of leaky gut syndrome ( LGS), allergy and inflammatory conditions in the treated children. Thereby, a dietary treatment with the compounds of the invention would work as

[0210] - prophylactic treatment for reducing the likelihood of development of severe symptoms of LGS, like diarrhea, bloating, and fatigue, in said young children, but also consequences of bacterial and viral infections of their GIT, such as allergic and inflammatory conditions, and malnutrition problems; or / and dietary enhancement of therapy used for young children diagnosed with a GIT infection, an allergic or inflammatory condition or disease, such as colic, pediatric inflammatory bowel disease or necrotizing enterocolitis (NEC), by supporting the young child healthy body development and maturation.

[0211] Accordingly, one aspect of the invention relates to a synthetic compound of formula (1), or salts thereof, or composition essentially consisting thereof: wherein

[0212] R1is a C13-C17 alkyl chain;

[0213] R2is a C15-C19 alkyl chain; and

[0214] R3is a carbohydrate moiety selected from:

[0215] Neu5Aca2-3-Gaipi-4-Glcpi-, or

[0216] Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-. for use in dietary treating of a young child, wherein said composition when administered for at least 14 consequent days increases the total amount of amount of SCFA in the GIT of said young child by at least 5%, and wherein said dietary treating restores, maintains or improves intestinal tissue structure and mucus production by the intestinal goblet cells, improves the mucus quality and mucosal immune and inflammatory status, and gut permeability in said young child.

[0217] In one preferred embodiment, the synthetic glycolipid is a compound of formula (1), wherein both R1and R2(independently) are C13-C17 alkyl chain.

[0218] One of the beneficial effects associated with an enhanced production of SCFA by the host microbiota in the gut is that SCFAs activate anti-inflammatory pathways of the host (i.e. young child). These effects, all, results on rection of inflammation and restoration of normal gut functioning of the host (Akhar, M., et al (2022) Animal Nutrition 8:350-360; https: / / doi.Org / 10.1016 / i.aninu.2021.ll.005).

[0219] Accordingly, the composition of the present invention can be used to reduce inflammation in the GIT, in particular the small intestine, of a young child. This can be accomplished by administering to the young child an effective amount of the said composition as defined above. In an embodiment, the composition can be used to reduce lipopolysaccharide induced inflammation in the gastro-intestinal tract of a young child by administering to the young child an effective amount of the composition as defined above.

[0220] The anti-inflammatory effect of the compounds or compositions described herein can be effectively used as dietary prophylactic or enhancing therapy means for development of such pediatric inflammatory diseases as, e.g.,

[0221] • Ulcerative colitis. A disease of the large intestine (colon) that causes inflammation and ulcers that can produce pus, bleeding, and mucus. The most common symptoms are bloody stools or diarrhea and abdominal cramping, but it can also cause weight loss, fatigue, anemia, skin lesions, joint pains, rashes, and other symptoms. It most commonly affects teenagers but can also be seen in infants and very young children.

[0222] • Very early onset IBD (VEO-IBD). A term used for IBD when it is diagnosed in children under 6 years of age. In infants, it is referred to as infantile IBD. The symptoms can be similar to IBD in an older child or adult, though they tend to have involvement of the colon more frequently.

[0223] • Lymphocytic colitis. A form of IBD where specific immune cells called “lymphocytes” are seen in the inner lining of the large intestine (colon). The most common symptom is diarrhea.

[0224] • Eosinophilic gastroenteritis (EGID). A digestive inflammatory disease that may or may not evolve into IBD over time. It can also have similar symptoms including abdominal pain, occasional diarrhea, constipation, nausea, and vomiting.

[0225] Accordingly, in one embodiment, compositions essentially consisting of or comprising compounds of the inventions described above, a child nutritional formula, can be used for dietary treatment of a young child, wherein an effective amount of said composition or child nutritional formula administered to said young child orally in one or more dosages during at least 14

[0226] - restores, maintains and / or improve said young child gut epithelium structure and functioning, and / or

[0227] - reduces the symptoms of leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease selected from abdominal pain, diarrhea, fatigue; or

[0228] - reduce the likelihood of development of a health disorder in said young child, wherein said health disorder is selected from leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease. The compounds, compositions and child nutritional formula of the invention are useful as dietary means both for healthy young children, like infants and toddlers, and those young children that experience health disorders. According to the invention, both groups of young children would benefit from use of the inventive compounds and compositions described herein. According to the invention, the compounds and compositions can help to reduce systemic and gastro-intestinal inflammation, in particular, and alleviate the associated symptoms, such as abdominal pain, diarrhea, ulcers, fatigue , etc, in young children experiencing thereof. However, the compositions described herein are not therapeutic, but nutritional, and they are intended for use in dietary management of these symptoms in young children. The compositions may be used as dietary enhancement of therapeutic treatment prescribed for curing these health conditions and symptom alleviation. The compounds and compositions can also dietary prevent or reduce the likelihood of occurrence of health conditions leading to these symptoms, like those health conditions that are described above, in healthy young children. Since a 14-day dietary consumption of the compounds, compositions or child nutritional formula of the invention increases the amount of SCFA in the young child GIT, and an increased amount of SCFA helps to reduce inflammation in the GIT of the young child and reduce the GIT inflammation-associated symptoms, like abdominal pain, diarrhea, ulcers, fatigue, in one embodiment, the invention relates to a non-therapeutic use of compounds, compositions or child nutritional formula of the invention, for increasing the amount of SCFA, specifically, acetate and butyrate, in the GIT of a young child, wherein an effective amount of said compound, salt, composition or child nutritional formula is administered orally to said young child daily in one or dosages during at least 14 days, In one embodiment, the compounds, compositions or child nutritional formula are used for in reducing bacterial or viral pathogen indued inflammation in the GIT, in particular, lipopolysaccharide induced inflammation in the gastro-intestinal tract

[0229] The effective amounts of the composition required to be administered will vary depending upon factors such as the risk and severity of the inflammation, any underlying medical condition or disease, age, the delivery form of the composition, and other medications being administered. However, the required amount can be readily set by a medical practitioner and would generally be in the range from about 1 mg to about 30 mg per day, in certain embodiments from about 2 mg to about 20 mg per day, for example from about 3 mg to about 10 mg per day. An appropriate dose can be determined based on several factors, including, for example, body weight and / or condition, the severity of the inflammation, other ailments and / or diseases, the incidence and / or severity of side effects and the manner of administration. Appropriate dose ranges may be determined by methods known to those skilled in the art. The effective amounts of individual compounds of compositions of the invention are discussed in detail above. The duration of the administration of the composition will vary depending upon factors such as the risk and severity of the inflammation, child age, the form of the composition, the dose and other medications being administered. However, the duration can be readily set by a medical practitioner. For example, the composition can be administered during the first week of life of the child, 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 three years of life. The composition can be administered daily, or with intervals between administration longer than a day. Further, the composition can be administered more than once a day. Preferably, the compounds and compositions are administered in effective amounts to a subject-in-need for at least 14 consequent days in one or more dosages per day. It is advantageous to administer the compounds or compositions of the invention in a form of a child nutritional formula as any of the described herein.

[0230] Some preferred, but non-limiting, embodiments of the invention are summarized below.

[0231] 1. A composition comprising one or more compounds of formula (1), or salts thereof: wherein

[0232] R1is a C13-C17 alkyl chain;

[0233] R2is a C15-C19 alkyl chain; and

[0234] R3is a carbohydrate moiety selected from:

[0235] Neu5Aca2-3-Gaipi-4-Glcpi-, or

[0236] Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-.

[0237] 2. A composition comprising a compound of formula (2), or a salt thereof: wherein, R1and R2are defined as for the compound of formula (1).

[0238] A composition comprising a compound of formula (2) and of formula (3), or salts thereof: wherein R1and R2are as defined as for the glycosphingolipid of formula (1).

[0239] A composition according to 3, wherein the ratio between the compound of formula (2) and the compound of formula (3) is from about 0.2:1 to about 14:1, preferably from about 0.2:1 to 5:1.

[0240] A composition according to any one of 1 to 4, further comprising a compound of formula (4): wherein R1and R2are defined as for the compound of formula (1).

[0241] A composition according to any one of 1 or 5, further comprising a compound of formula

[0242] (5): (5) wherein R1and R2are defined as for the compound of formula (1).

[0243] 7. A composition according to any one of 3 to 6, further comprising a compound of formula (6):

[0244] O R2^NH Galpl-4-Glcpl-O^A ^^R1 wherein R1and R2, Glc, Gal, and Neu5Ac are defined as for the compound of formula (1).

[0245] 8. A child nutritional formula comprising a composition of any one of claims 1 to 6.

[0246] 9. A child nutritional formula according to 8, wherein the child nutritional formula comprising a composition according to 3 or 4, wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

[0247] 10. A child nutritional formula according to 8, wherein the child nutritional formula comprising a composition according to 2, wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

[0248] 11. A child nutritional formula according to any one of 8 to 10, for use in reducing inflammation in the gastro-intestinal tract of a child.

[0249] 12. A child nutritional formula of any one of 8 to 10, for use in reducing lipopolysaccharide induced inflammation in the gastro-intestinal tract.

[0250] 13. A child nutritional formula of any one of 8 to 10, for use in reducing inflammation in the small intestine of a child.

[0251] Further preferred, but non-limiting, embodiments of the invention include.

[0252] 1. A composition essentially consisting of one or more compounds of formula (1), or salts thereof: wherein

[0253] R1is a C13-C17 alkyl chain, preferably, Ci5or Ci7alkyl chain;

[0254] R2is a C15-C19 alkyl chain, preferably, Ci5or Ci7alkyl chain; and

[0255] R3is a carbohydrate moiety selected from:

[0256] Neu5Aca2-3-Gaipi-4-Glcpi-, or

[0257] Neu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-. A composition essentially consisting of a compound of formula (2), or a salt thereof: wherein, R1and R2are defined as for the compound of formula (1). A composition essentially consisting of mixture of a compound of formula (2) and of formula

[0258] (3), or salts thereof:

[0259] (3), wherein R1and R2are as defined as for the compound of formula (1). A composition essentially consisting of mixture of a compound of formula (2) and of formula (3), or salts thereof, wherein the ratio between the compound of formula (2) and the compound of formula (3) is from about 0.2:1 to about 14:1, preferably from about 0.2:1 to 5:1. A composition according to (1) (2), (3) or (4), further comprising a compound of formula (4): wherein R1and R2are defined as for the compound of formula (1). A composition according to (1) (2), (3), (4) or (5), further comprising a compound of formula (5): wherein R1and R2are defined as for the compound of formula (1). A composition according to (1) (2), (3), (4), (5) or (6), further comprising a compound of formula (6):

[0260] O

[0261] R2^NH

[0262] Gaipi-4-Glcpi-O.^L ^^R1I OH wherein R1and R2, Glc, Gal, and Neu5Ac are defined as for the compound of formula (1). 8. A child nutritional formula comprising a compositions (1) (2), (3), (4), (5), (6) or (7) .

[0263] 9. A child nutritional formula comprising a composition (2) or (3), wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

[0264] 10. A child nutritional formula comprising a composition (2), wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

[0265] 11. A child nutritional formula comprising a composition (1) (2), (3), (4), (5), (6) or (7), for use in reducing inflammation in the gastro-intestinal tract of a child.

[0266] 12. A child nutritional formula comprising a compositions (1) (2), (3), (4), (5), (6) or (7), for use in reducing lipopolysaccharide induced inflammation in the gastro-intestinal tract.

[0267] 13. Non-therapeutic use of a compound of formula (1), a salt thereof, or a composition (1) (2), (3), (4), (5), (6) or (7), or child nutritional formula (8), (9), (10) or (11), for increasing the amount of short chain fatty acids, preferably acetate and / or butyrate, in the gastrointestinal tract of a young child, wherein an effective amount of said compound, salt, composition or child nutritional formula is administered orally to said young child daily in one or dosages during at least 14 days.

[0268] 14. A composition (1) (2), (3), (4), (5), (6) or (7), or a child nutritional formula (8), (9), (10) or (11), for use in dietary treatment of a young child, wherein an effective amount of said composition or child nutritional formula administered to said young child orally in one or more dosages during at least 14 days

[0269] - restores, maintains and / or improve said young child gut epithelium structure and functioning, and / or

[0270] - reduces the symptoms of leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease selected from abdominal pain, diarrhea, fatigue; or

[0271] - reduce the likelihood of development of a health disorder in said young child, wherein said health disorder is selected from leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease.

[0272] Compositions of the above embodiments are synthetic compositions.

[0273] All compounds of formulas (1), (2), (3), (4), (5), and (6) are synthetic compounds.

[0274] Examples The examples described below are for illustration purposes only. The invention is not limited in scope to embodiments described in the examples.

[0275] Example 1: General procedure for the production of a composition comprising a compound of formula (8)

[0276] The 2,3-trans-sialidase catalyzed sialylation reaction is performed in an aqueous solution at a pH between about 6.0 to about 7.0. A typical reaction mixture contains the compound of formula (7) (1 eq.), 3'-sialyllactose (3.5 eq.), p-cyclodextrin (0.5 eq.), and 2,3-transialidase (TcTS, 0.4 g / L). The reaction mixture is stirred for about 3 to 6 hours at a temperature of about 37°C and then the - galactosidase is added (0.13 g / L). The reaction mixture is nanofiltered using 300 to 500 Da membranes, a pressure of 15 to 20 bar and a temperature of about 30-40°C for about 6 to 8 hours. The nanofiltration retentate (NFR) is heated to a temperature between about 60 to 95°C for about 10 to 60 minutes. The NFR is then diafiltered (DF) using 250 kDa membranes, a pressure of around 15 to 20 bar, a temperature of about 15 to 40°C, and around 10 diafiltration volumes relative to the volume of the NFR.

[0277] Example 2: General procedure for the production of a composition comprising a compound of formula (8) and of formula (10)

[0278] The enzymatic reaction is performed in an aqueous solution at a pH between about 6.5 to about 7.5, and at a temperature of about 37°C. A typical reaction mixture contains the compound of formula (8) (1 eq.), / V-acetylneuraminic acid (Neu5Ac, 2.5 eq.), P-cyclodextrin (0.5 25 eq.), ATP (3.5 eq.), CMP (0.27 eq.), MgCL (20 mM), and the following enzymes: sialyltransferase (CST-II I53A, 5g / L), cytidine monophosphate kinase (MtCMK, 12g / L), nucleoside diphosphate kinase (MtNDK, 6g / L), N- acylneuraminate cytidyltransferase (NmCSS, lg / L), inorganic diphosphatase (EcPPase, 2.5 pL / mL).

[0279] Example 3: General procedure for the production of a composition comprising a compound of formula (2) or compounds of formula (2) and (3)

[0280] A composition comprising a compound of formula (8), or compounds of formula (8) and (10) is dissolved in methanol, and the triazine-based acylating agent is added. The resulting suspension is stirred at 50°C until a thin layer chromatography (TLC) analysis shows complete consumption of the starting material. Then, water is added, and the reaction mixture is cooled down to room temperature. A solid precipitates, which is filtered and dried in vacuum to obtain the final product.

[0281] Example 4: Production of a composition comprising GM3

[0282] The composition comprising GM3 is synthesized in two steps. In the first step a composition comprising / V-lyso-GM3 is obtained staring from lactosyl-D-erythro-sphingosine as described in Example 1. In the second step the composition comprising / V-lyso-GM3 is subjected to / V-acylation with the triazine-based acylating agent as described in Example (3).

[0283] The triazine-based acylating agent is 4,6-dimethoxy-l,3,5-triazyn-2-yl stearate.

[0284] The composition comprising GM3, produced by the process above, contains about 75-85 wt. % of ganglioside GM3 [a- / V-acetylneuraminosyl-(2->3)-O-p-D-galactopyranosyl-(l->4)-p-D-glucopyranosyl- (l^l')- / V-stearoyl-D-eryt / iro-sphingosine), about 5-10 wt.% by mass of LacCer[N(18) S( 18)] [P-D- galactopyranosyl-(l->4)-p-D-glucopyranosyl-(l->r)- / V-stearoyl-D-erythro-sphingosine]; and about 0.4-1 wt.% of GlcCer[N(18) S( 18)] [p-D-glucopyranosyl-(l^l')-A / -stearoyl-d-eryt / iro-sphingosine]. The GM3, LacCer and GlcCer all contain a Cis fatty acid acylated to the sphingosine base.

[0285] Example 5: Production of a composition comprising GM3 and GD3

[0286] The composition comprising GM3 and GD3 is synthesized in three steps. In the first step a composition comprising / V-lyso-GM3 is obtained as described in Example 1. In the second step a composition comprising / V-lyso-GM3 and / V-lyso-GD3 is obtained as described in Example 2. In the third step the composition comprising / V-lyso-GM3 and / V-lyso-GD3 is subjected to / V-acylation with the triazine-based acylating agent as described in Example 3.

[0287] The composition comprising GM3 and GD3, produced by the process above, contains about 12-60 wt. % of ganglioside GM3 [a- / V-acetylneuraminosyl-(2->3)-O-p-D-galactopyranosyl-(l->4)-p-D- glucopyranosyl-(l->r)- / V-stearoyl-D-erythro-sphingosine]; 20-55 wt. % of ganglioside GD3 [a- / V- acetylneuraminosyl-(2->8)-a- / V-acetylneuraminosyl-(2->3)-O-p-D-galactopyranosyl-(l->4)-p-D- glucopyranosyl-(l->r)- / V-stearoyl-D-erythro-sphingosine]; about 0-5 wt. % of ganglioside GT3 [a- / V- acetylneuraminosyl-(2->8)-a- / V-acetylneuraminosyl-(2->8)-a- / V-acetylneuraminosyl-(2->3)-O-p-D- galactopyranosyl-(l->4)-p-D-glucopyranosyl-(l->l')- / V-stearoyl-D-erythro-sphingosine]; about 5-10 wt.%> of LacCer[N(18) S( 18)] [p-D-galactopyranosyl-(l->4)-p-D-glucopyranosyl-(l->l')- / V-stearoyl-D- erythro-sphingosine]; and about 0.4-1 wt.%> of GlcCer[N(18) S(18)] [p-D-glucopyranosyl-(l->l')- / V- stearoyl-d-erythro-sphingosine]. The GM3, GD3, GT3 LacCer and GlcCer all contain a Cis fatty acid acylated to the sphingosine base.

[0288] Example 6. Production of Enzymes

[0289] The enzymes used for the production of the composition as described above are either purchased from commercial manufacturers or produced in-house by standard molecular cloning techniques using recombinant E. coli as the host.

[0290] Example 7. Study of the impact on metabolite production of healthy infants by the compositions of the present invention. The objective of the study was to investigate the impact on metabolite production of healthy, exclusively formula-fed infants (1-3 months) by the different ganglioside compositions of the present invention. Three different ganglioside compositions were tested (test products), particularly one composition comprising GM3 (referred to as test product GM3), and two compositions comprising GM3 and GD3 (referred to as test product GM3-GD3 mix 1 and test product GM3-GD3 mix 2), wherein test products GM3-GD3 mix 1 and GM3-GD3 mix 2 were characterized by a different ratio of the two gangliosides. Particularly, GM3-GD3 mix 1 is characterized by a mass ratio between GM3 and GD3 of about 1:4, whereas GM3-GD3 mix 2 is characterized by a mass ratio between GM3 and GD3 of about 3:1.

[0291] The three test products were tested as such or upon being subjected to gastric and small intestinal digestion / absorption prior to colonic incubation. The digested test products are referred to as GM3+, GM3-GD3 mix 1+, and GM3-GD3 mix 2+. Tests were performed using the ex vivo SIFR® technology, wherein the impact on metabolite production such as production of SCFA, lactate and bCFA, and key fermentation parameters such as pH and gas were assessed for 6 test subjects. The test products were compared against a no-substrate control, while predigested products and their undigested counterparts were also compared against each other.

[0292] Test Study:

[0293] Test products GM3, GM3-GD3 mix 1, GM3-GD3 mix 2, and their digested forms were tested at 1 g / L against a no-substrate control (NSC). The dose was determined based on a previous study showing that only minor effects on Bifidobacterium species were observed at 0.5 g / L, while dosing gangliosides at > 1 g / L exerted pronounced effects. The test products were dosed as such or upon pre- digestion prior to colonic incubation. Test products, sample type, their corresponding compositions and doses are summarized in Table 1.

[0294] Table 1. Test products, Sample Type, Compositions and Doses

[0295] *Abbreviations used in the schematic presentation of the study shown in Figure 1

[0296] The digested test products were subjected to gastric and small intestinal digestion procedures according to the method adapted to the infant gut published by Menard et al., Food Chem. 2018, 240, 338-345.

[0297] Subsequently, small intestinal absorption was stimulated by means of a 1.0 kDa dialysis membranes resulting in absorption of gangliosides (size = 1.6 kDa) only upon their digestion to fractions <1.0 kDa.

[0298] The colonic fermentation of the test products by the infant gut microbiota was investigated using the SIFR® technology (Cryptobiotix). The study design is schematically presented in Figure 1. An ex vivo SIFR® study was implemented, simulating colonic fermentation of test products by the gut microbiota derived from healthy infants (n = 6).

[0299] An ex vivo SIFR® study was implemented, simulating colonic fermentation of test products by the gut microbiota derived from healthy infants (n = 6).

[0300] Faecal samples were collected according to a procedure approved by Ethics Committee of the University Hospital Ghent (reference number BC-09977). The selection criteria for the donors were as follows: healthy infants of 1-3 months. The exclusion criteria were breastfeeding for more than 1 month in the past and 2 weeks prior to faecal donation, antibiotic use in 30 days before sample delivery for the study and previous NEC or gut surgery. This resulted in the enrolment of 6 specific test subjects (Table 2). The average age of the infants was 2.2 (± 1.0) months.

[0301] The simulation parameters were as follows:

[0302] • Test Product = 7

[0303] • No substrate control (NSC) = background medium + microbiota (no product)

[0304] • Undigested products: GM3, GM3-GD3 mix 1, GM3-GD3 mix 2 • Predigested products: GM3 +, GM3-GD3 mix 1 +, GM3-GD3 mix 2 +

[0305] • Time points = Oh (only for NSC), 24 h (all test products)

[0306] Key fermentation parameters and metabolite production were measured at 0 h (only NSC) and 24h (for all test products).

[0307] During the current study, acetate, propionate, butyrate, valerate and branched chain fatty acids (bCFAs) (sum of isobutyrate, isovalerate and isocaproate) were determined via a GC-FID approach. Lactate was measured with a high-throughput spectrophotometric method. Further, pH and gas production were also measured as key markers for microbial metabolic activity.

[0308] For exploratory evaluation of the obtained results, a series of principle component analysis (PCA) was performed. The two principal components with the largest eigenvalues were plotted.

[0309] For the statistical evaluation of the treatment effects on key fermentation parameters, cell counts, microbial diversity (4 indices) and microbial composition (phylum level) across 6 different donors, a repeated measures ANOVA analysis was performed (~ based on paired t-testing, thus accounting for fact that values are compared between samples of a given donor). The statistical significance of the potential treatment effects was determined via Benjamini-Hochberg post hoc testing30.

[0310] Test Study Results

[0311] • Undigested test product (GM3-GD3 mix 1, GM3 and GM3-GD3 mix 2)

[0312] The treatment of the undigested GM3-GD3 mix 1, GM3 and GM3-GD3 mix 2 resulted in the following key observations:

[0313] - pH Tended to decrease (not significant), while gas production tended to increase, most significantly for GD3 (+28%).

[0314] - Significant increase in total SCFA production, with the highest increase observed for GM3- GD3 mix 1 (+14.7%), followed by GM3-GD3 mix 2 (+10.6%) and GM3 (+6.0%).

[0315] This is largely attributed to a significant increase in acetate production for GM3-GD3 mix 1 (+19.0%), GM3-GD3 mix 2 (+14.2%) and GM3 (+8.1%).

[0316] - Propionate levels tended to decrease (-3.9% for GM3-GD3 mix 1, -2.0% for GM3-GD3 mix 2 and -0.6% for GM3), while butyrate levels tended to increase (+28.2% for GM3-GD3 mix 1, +12.5% for MIX and +7.1% for GM3), bCFA significantly decreased, with the strongest decrease observed for GM3 (-8.3%), followed by GM3-GD3 mix 1 (-7.5%) and GM3-GD3 mix 1 (-5.6%).

[0317] • Predigested test products (GD3+, GM3+ AND MIX+) Compared to the undigested counterpart (GM3-GD3 mix 1, GM3 and GM3-GD3 mix 2), the treatment of the predigested GM3-GD3 mix 1+, GM3+ and GM3-GD3 mix 2+ resulted in the following key observations:

[0318] Stronger decrease in pH with lower increase in gas production.

[0319] - Lower increase in total SCFA production: GM3-GD3 mix 1+ (+11.6%), GM3-GD3 mix 2+ (+7.1%) and GM3+ (+5.9%).

[0320] - Similarly, lower increase in acetate production was observed for : GM3-GD3 mix 1+ (+17.1%), GM3-GD3 mix 2+ (12.1%) while higher increase observed for GM3 (+10.7%)

[0321] Stronger decrease in propionate, especially for GM3-GD3 mix 2+ (-8.1%), followed by GM3- GD3 mix 1+ (-6.2%) and GM3+ (-5.1%).

[0322] - Lower increase in butyrate: GM3-GD3 mix 1+ (+19.8%), GM3-GD3 mix 2+ (+5,9%), GM3+ (+1.0%)

[0323] - Significantly stronger decrease in bCFA: GM3+ (-36.3%), GM3-GD3 mix 1+ (-28.4%) and GM3- GD3 mix 2+ (-22.0%)

[0324] Discussion & Conclusion

[0325] Upon simulating a single intake, the gangliosides impacted microbial metabolite production, i.e. decreased pH and increased gas production, significantly increased acetate levels, while tending to increase butyrate levels (thus also significantly increasing total SCFA). These stimulatory effects were the most and the least pronounced for GM3-GD3 mix 1 / GM3-GD3 mix 1+ and GM3 / GM3+, respectively, while GM3-GD3 mix 2 / GM3-GD3 mix 2+ exerted intermediate effects.

[0326] Gut bacteria such as Bifidobacterium spp. present in infant's gut are capable of degrading milk gangliosides and releasing sialic acid. Sialic acid is a functional sugar that is used as a fermentation substrate by a number of gut bacteria, many of which promote SCFA production. Since GM3-GD3 mix 1 has a higher content of GD3 which contains more sialic acid residues compared to GM3 per unit mass (2 vs 1), GM3-GD3 mix 1 could provide more nutrients for microbial fermentation, especially by SCFA-producing bacteria.

[0327] In addition, the stimulatory effects on acetate and butyrate (thus, total SCFA) were only slightly reduced when gangliosides were subjected to upper GIT passage, e.g., +11.6% more total SCFA for GM3-GD3 mix 1+ compared to +14.7% for GM3-GD3 mix 1. This suggests that gastric and small intestinal digestion has only a minor impact on the stimulatory effects on gangliosides on health related SCFAs production. Interestingly, all test products tended to decrease propionate and bCFA production. In contrast to the stimulatory effects on acetate and butyrate, these inhibitory effects of the tested products are significantly enhanced by simulated oral, gastric, and small intestinal digestion. For example, bCFAs production decreased of -8.3% on average for undigested GM3 while decreased -36.3% for predigested GM3+.

[0328] Overall, the tested products strongly stimulated acetate and butyrate levels, thus increasing total SCFA production. The strength of these stimulatory effects ranked as follows: GM3-GD3 mix 1 > GM3-GD3 mix 2 > GM3. These effects were only mildly reduced by the pre-digestion prior to colonic incubation. In contrast, propionate and bCFAs production decreased in response to all test products and were further lowered by the pre-digestion of the test products when compared to the undigested counterparts.

[0329] Overall, the obtained results suggest that the compositions of the present invention impact microbial metabolite production. Besides increasing acetate / butyrate production, the compositions of the present invention lowered propionate / bCFAs levels, potentially resulting in a series of health benefits for the human host. Pre-digestion only had minor effects on the increase of acetate / butyrate levels, while significantly further decreasing propionate / bCFA levels.

[0330] Example 8. Reduction of LPS induced inflammation

[0331] Purified and stimulated human monocytes are prepared from heparinized, fresh, human peripheral blood by first diluting the blood to 2x volume with a cold (4°C) endotoxin-free, phosphate-buffered saline (PBS) containing 1 pg / ml polymyxin B. The diluted blood is overlaid on a cold (4°C) lymphocyte separation solution containing 1 pg / ml polymyxin B and centrifuged at 800 g for 25 min at 4°C. The peripheral blood mononuclear cell (PBMC) fraction is collected and diluted to 2x volume of a wash solution at pH 7.5 (PBS, 1% heat-inactivated faecal bovine serum (FCS), 5 mM EDTA, 1 pg / ml polymyxin B).

[0332] The PBMCs are separated by centrifugation at 600 g for 10 min at 4°C, washed twice, resuspended in 750 pl of the wash solution and incubated with 120 pl anti-human CD14 magnetic particles for 30 minutes at room temperature. CD14-positive monocytes are separated by magnetic field and washed 3x with the wash solution. The resulting purified cells are resuspended in a cold, low-glucose Dulbecco's Modified Eagle Medium (DMEM) with 0.75% FCS and left on ice for 45 minutes. Thereafter the purified and stimulated monocytes are counted, diluted to 2.0 x 105cells / ml with a culture medium (low-glucose DMEM, 0.75% FCS, 40 ng / ml recombinant human granulocytemacrophage colony-stimulating factor), and cultured in 96-well plates (100 pl / well) overnight at 37°C under 5% CO2 atmosphere. The composition comprising ganglioside GM3 having a Cis acyl moiety (prepared according to example 4), ganglioside GM3 having C22 and ganglioside GM3 having a C24 acyl moieties (synthesized as described by Mauri et al, Glycoconjugate J. 1999, 16, 197 and Murase et al., Carbohydrate Res. 1989, 188, 71) were dissolved in warm low-glucose DMEM, to obtained three solutions having 0.5 nM concentration. The solutions are stored at -30°C and then diluted to 100 pM concentration using low-glucose DMEM just prior to use.

[0333] Activated monocytes are separately primed for 30 minutes with the solutions containing the composition comprising GM3 having a Cis acyl moiety, and the solutions of ganglioside GM3 having a C22 acyl moiety and of ganglioside GM3 having a C24 acyl moiety, then stimulated with LPS. As control, activated monocytes are stimulated with LPS. After 18 hours of culturing, the media is collected and subjected to ELISA using ELISA kits for human IL-6 and human TNF-a.

[0334] By themselves (i.e., without LPS stimulation), the composition comprising ganglioside GM3 having a Ci8 acyl moiety, the ganglioside GM3 having C22 and the ganglioside GM3 having a C24 acyl moieties do not induce IL-6 and TNF-a production. Upon LPS stimulation, the control exhibits significant IL-6 and TNF-a production. The IL-6 and TNF-a production is amplified with GM3 having a C22 acyl moiety and GM3 a having C24 acyl moiety. However, the composition comprising ganglioside GM3 having a Ci8 acyl moiety reduces IL-6 and TNF-a production.

[0335] LPS is present in the intestine of infants. Including the composition of the present invention in the diet of the infants has the potential to reduce LPS-induced inflammation. Cow's milk GM3 (which mainly comprise GM3 having a C22 and ganglioside GM3 having a C24 acyl moieties) has the potential to amplify LPS-induced inflammation.

[0336] Example 9. Child nutritional formula

[0337] A child nutritional formula having the following composition is prepared:

[0338] The disclosure should not be seen in any way restricted to the embodiments described and a person with ordinary skill in the art will foresee many possibilities to modifications thereof.

[0339] The above described embodiments are combinable. The following claims further set out particular embodiments of the disclosure.

Claims

CLAIMS1. A composition comprising one or more compounds of formula (1), or salts thereof:whereinR1is a C13-C17 alkyl chain, preferably a C13 alkyl chain;R2is a C15-C19 alkyl chain, preferably, C15 or C17 alkyl chain; andR3is a carbohydrate moiety selected from:Neu5Aca2-3-Gaipi-4-Glcpi-, orNeu5Aca2-8-Neu5Aca2-3-Gaipi-4-Glc pi-.

2. The composition according to claim 1, comprising a compound of formula (2), or a salt thereof:(2), wherein, R1and R2are defined as for the compound of formula (1).

3. The composition according to claim 1, comprising a mixture of a compound of formula (2) and of formula (3), or salts thereof:(2),(3), wherein R1and R2are as defined as for the glycosphingolipid of formula (1).

4. The composition according to claim 3, wherein the ratio between the compound of formula (2) and the compound of formula (3) is from about 0.2:1 to about 14:1, preferably from about 0.2:1 to 5:1.

5. The composition according to any one of claims 1 to 4, wherein the compounds of formula (l)-(3), are synthetic compounds.

6. The composition according to claim 1, essentially consisting of a compound of formula (1).

7. The composition according to claim 6, essentially consisting of a compound of formula (2).

8. The composition according to claim 7 , essentially consisting of a mixture of a compound of formula (2) and a compound of formula (3).

9. The composition according to any one of claims 1 to 8, further comprising a compound of formula (4):wherein R1and R2are defined as for the compound of formula (1).

10. The composition according to any one of claims 1 to 9, further comprising a compound of formula (5):Glcpi-(5) wherein R1and R2are defined as for the compound of formula (1).

11. The composition according to any one of claims 3 to 10, further comprising a compound of formula (6):OR2^NHGalpl-4-Glcpl-O^^ ^^R1wherein R1and R2, Glc, Gal, and Neu5Ac are defined as for the compound of formula (1).

12. A child nutritional formula comprising a composition of any one of claims 1 to 11.

13. The child nutritional formula according to claim 12, wherein the child nutritional formula comprising a composition according to claims 3 or 4, wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

14. The child nutritional formula according to claim 12, wherein the child nutritional formula comprising a composition according to claim 2, wherein the total content of said composition in said child nutritional formula is between about 5 mg / l and about 35mg / l, preferably between about 6 mg / l and about 26 mg / l.

15. The child nutritional formula according to any one of claims 12 to 14, for use in reducing inflammation in the gastro-intestinal tract of a young child.

16. The child nutritional formula of any one of claims 12 to 14, for use in reducing lipopolysaccharide induced inflammation in the gastro-intestinal tract.

17. The child nutritional formula of any one of claims 12 to 14, for use in reducing inflammation in the small intestine of a young child.

18. Non-therapeutic use of a compound of formula (1), a salt thereof, or a composition of any of claims 1 to 11, or child nutritional formula of any of claims 12-14, for increasing the amount of short chain fatty acids, specifically, acetate and / or butyrate, in the gastrointestinal tract of a young child, wherein an effective amount of said compound, salt, composition or childnutritional formula is administered orally to said young child daily in one or dosages during at least 14 days.

19. A composition according to any of claims 1 to 11, or a child nutritional formula according any of claims 12-14, for use in dietary treatment of a young child, wherein an effective amount of said composition or child nutritional formula administered to said young child orally in one or more dosages during at least 14 days,- restores, maintains and / or improve said young child gut epithelium structure and functioning, and / or- reduces the symptoms of leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease selected from abdominal pain, diarrhea, fatigue; or- reduce the likelihood of development of a health disorder in said young child, wherein said health disorder is selected from leaky gut syndrome, allergy, colic, necrotizing enterocolitis, IBD or infectious disease.

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