Lutein and 1,3-dioleoyl-2-palmitoylglycerol for use in the prevention or treatment of gastrointestinal disorders

Abstract

The present invention relates to a combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol.

Description

Technical Field

[0001] This invention belongs to the field of nutrition, and more particularly relates to compositions for use in the prevention or treatment of gastrointestinal disorders in human subjects, particularly gastrointestinal disorders associated with reduced lactase expression. Background Technology

[0002] Gastrointestinal disorders are common in humans, especially in Western countries where high-fat diets are prevalent. It is estimated that 60 to 70 million Americans suffer from gastrointestinal problems.

[0003] Gastrointestinal problems are commonly associated with lactose intolerance, which affects 57% to 65% of the global population. Lactose intolerance is caused by reduced or absent activity of the intestinal enzyme lactase—phlorizin hydrolase (more commonly known as lactase), which is responsible for digesting lactose. Reduced or absent lactase activity leads to increased osmotic load in the small intestine and fermentation of lactose by bacterial flora, which may result in increased production of short-chain fatty acids and gas. This can subsequently cause abdominal pain, diarrhea, and bloating.

[0004] A common cause of lactose malabsorption is lactase non-persistence, a condition characterized by reduced lactase expression in infancy. In contrast, congenital lactase deficiency (a rare condition where newborns exhibit severe symptoms) results from a complete lack of lactase. Furthermore, lactose malabsorption can be secondary to acquired conditions, including small intestinal bacterial overgrowth, infectious enteritis (i.e., giardiasis), or mucosal damage caused by celiac disease, inflammatory bowel disease, medications, gastrointestinal surgery, short bowel syndrome, or radiation enteritis, leading to decreased carbohydrate absorption or downregulated lactase expression in the small intestine.

[0005] Furthermore, the intestinal lactase activity of preterm infants has not yet reached a mature level. Early studies have shown that although intestinal lactase activity begins to develop in early pregnancy, the period of greatest increase in lactase occurs in late pregnancy (Antonowicz et al., Development and distribution of lysosomal enzymes and disaccharidases in human fetal intestine, Gastroenterology, 1974, 67, 51-58). At the same time, the digestion of lactose in preterm infants appears to be incomplete (Shulman et al., Absorption of lactose, glucose polymer, or combination in premature infants, J Pediatr, 1995, 127(4), 626-631).

[0006] Therefore, there is a need for compositions that prevent or treat gastrointestinal conditions associated with lactose malabsorption and / or reduced lactase expression. Summary of the Invention

[0007] The inventors unexpectedly discovered that the combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) increases lactase expression in the gastrointestinal tract. Furthermore, the inventors also found that when used in combination with lutein and 1,3-dioleoyl-2-palmitoylglycerol, the presence of fermentation products of indigestible oligosaccharides (i.e., short-chain fatty acids (SCFAs)) synergistically increases lactase expression in the gastrointestinal tract.

[0008] Without being bound by theory, the inventors speculated that increasing lactase expression in the epithelial cells of subjects suffering from gastrointestinal symptoms due to impaired lactase expression might lead to increased lactose tolerance in the subjects, thereby alleviating gastrointestinal symptoms.

[0009] Therefore, the present invention relates to a combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) for use in the prevention or treatment of gastrointestinal disorders in subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract. Preferably, the combination is administered orally, and more preferably, a nutritional composition comprising the combination is administered orally. Attached Figure Description

[0010] Figure 1The effects of SCFA, lutein, OPO, lutein + OPO, or lutein + OPO + SCFA on lactase mRNA expression in Caco-2 cells were shown compared to the negative control.

[0011] List of preferred embodiments 1. An oral composition comprising lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) for use in the prevention or treatment of gastrointestinal disorders in human subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract.

[0012] 2. The composition for use according to Example 1, wherein the prevention or treatment of the gastrointestinal symptom is carried out by increasing the expression of lactase in the gastrointestinal tract.

[0013] 3. The composition for use according to Example 1 or 2, wherein the composition comprises an indigestible oligosaccharide.

[0014] 4. The composition for use according to Example 3, wherein the indigestible oligosaccharide is selected from galactooligosaccharides (GOS), fructooligosaccharides (FOS), human milk oligosaccharides (HMO), or combinations thereof.

[0015] 5. The composition for use according to any of the foregoing embodiments, wherein the impaired lactase expression is due to lactase non-persistence, preterm birth, celiac disease, inflammatory bowel disease, short bowel syndrome, intestinal infection, or exposure of the subject to chemotherapy or a combination thereof.

[0016] 6. The composition for use according to Example 5, wherein the impaired lactase expression is due to lactase non-persistence, prematurity, or a combination thereof.

[0017] 7. The composition for use according to any of the foregoing embodiments, wherein the symptoms of the gastrointestinal ailment are selected from diarrhea, constipation, flatulence, bloating, abdominal pain, or a combination thereof.

[0018] 8. The composition for use according to any of the foregoing embodiments, wherein the oral composition is a nutritional composition.

[0019] 9. The composition for use according to any of the foregoing embodiments, wherein the oral composition is infant formula, follow-up formula or toddler formula.

[0020] 10. The composition for use according to Example 9, wherein the oral composition is an infant formula for premature infants and / or small for gestational age infants.

[0021] 11. The composition for use according to any of the foregoing embodiments, wherein the composition comprises ≥ 5 μg lutein / 100 g dry weight of the composition.

[0022] 12. The composition for use according to any of the foregoing embodiments, wherein the composition comprises 0.5 to 15 g OPO / 100 g dry weight of the composition.

[0023] 13. The composition for use according to any of the foregoing embodiments, wherein the composition comprises 0.05-25 g of indigestible oligosaccharide per 100 g dry weight of the composition.

[0024] 14. The composition for use according to any of the foregoing embodiments, wherein the composition comprises 0-75 g lactose / 100 g dry weight of the composition.

[0025] 15. The composition for use according to any of the foregoing embodiments, wherein the composition comprises digestible carbohydrates, lipids, proteins and an energy content of at least 400 kcal, wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 5-1000 μg of lutein; b. 0.25-25 g of indigestible oligosaccharides, comprising fructooligosaccharides and galactooligosaccharides; and c.0.5-15 g 1,3-Dioleoyl-2-palmitoylglycerol. Detailed Implementation

[0026] A first aspect of the invention relates to a combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) for use in the prevention or treatment of gastrointestinal disorders in subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract. In a preferred embodiment, the combination is contained in an oral composition, more preferably a nutritional composition.

[0027] In some jurisdictions, the present invention can be defined as a method for preventing or treating a gastrointestinal symptom in a subject, wherein the gastrointestinal symptom is caused by impaired expression of lactase in the gastrointestinal tract, the method comprising orally administering to the subject a combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO), the combination preferably being contained in an oral composition.

[0028] In some jurisdictions, the invention may be defined as the use of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) in the manufacture of oral compositions, preferably nutritional compositions, for the prevention or treatment of gastrointestinal disorders in subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract.

[0029] In some jurisdictions, the administration of nutritional compositions to human subjects is considered non-therapeutic. In those cases, the invention can be described as a method comprising administering an oral nutritional composition, as defined above. For clarity, this method can also be defined as a non-therapeutic method. By definition, the term "non-therapeutic" excludes any therapeutic effect.

[0030] Preferably, the prevention or treatment of gastrointestinal disorders is carried out by increasing the expression of lactase in the gastrointestinal tract, and more preferably by increasing the expression of lactase in the small intestine.

[0031] The combination of 1,3-dioleoyl-2-palmitoylglycerol and lutein is at a therapeutically effective level.

[0032] Treatment group In a preferred embodiment, the human subject is an adult, preferably at least 50 years old, more preferably at least 65 years old.

[0033] In a preferred embodiment, the human subjects are selected from patients who have undergone abdominal surgery, patients who have experienced postoperative bowel dysfunction, and / or patients who are malnourished.

[0034] In another preferred embodiment, the human subject is an infant or toddler, preferably aged between 0 and 60 months, more preferably between 0 and 36 months. More preferably, the human subject is an infant. As used herein, the term "infant" refers to a human subject aged between 0 and 24 months, preferably between 0 and 12 months, and even more preferably between 0 and 6 months.

[0035] The infants are preferably preterm and / or small for gestational age (SGA) infants. Preterm infants are those born before the completion of the standard gestational period, i.e., before or after 37 weeks from the start of the mother's last menstrual period. SGA infants are those whose birth weight is below the 10th percentile of their gestational age. They are typically subjects of intrauterine growth restriction (IUGR). Preterm and / or SGA infants include low birth weight infants (LBW infants), very low birth weight infants (VLBW infants), and extremely low birth weight infants (ELBW infants). LBW infants are defined as those weighing less than 2500 g, VLBW infants as those weighing less than 1500 g, and ELBW infants as those weighing less than 1000 g.

[0036] Gastrointestinal diseases Gastrointestinal disorders are caused by impaired lactase expression in the gastrointestinal tract. Preferably, impaired lactase expression is due to lactase non-persistence, preterm birth, celiac disease, inflammatory bowel disease, short bowel syndrome, intestinal infection, or exposure to chemotherapy or a combination thereof. More preferably, impaired lactase expression is due to lactase non-persistence, preterm birth, or a combination thereof. Most preferably, impaired lactase expression is due to lactase non-persistence or preterm birth. In other words, preferably, a combination of lutein and OPO (and combinations included in oral compositions), preferably further combined with indigestible oligosaccharides, is used for the treatment of lactase non-persistence.

[0037] Preferably, the symptoms of the gastrointestinal disorder are selected from diarrhea, constipation, flatulence, bloating, abdominal pain, or combinations thereof. In other words, preferably, a combination of lutein and OPO (and combinations contained in oral compositions), preferably further combined with indigestible oligosaccharides, is used for the treatment of diarrhea, constipation, flatulence, bloating, abdominal pain, or combinations thereof.

[0038] In another preferred embodiment, the combination of lutein and OPO (and the combination contained in the oral composition), preferably further combined with indigestible oligosaccharides, is used for the treatment of diarrhea, constipation, flatulence, bloating, abdominal pain, or combinations thereof in preterm and / or small-for-gestational-age infants.

[0039] Lutein Preferably, lutein exists as free xanthophyll, xanthophyll ester, or other chemical forms of lutein. Lutein is abundant in leafy green vegetables such as spinach, kale, and yellow carrots. Lutein can be obtained or isolated by any method recognized by those skilled in the art. For example, lutein can be obtained by extraction from marigolds or other xanthophyll-rich sources, chemical synthesis, fermentation, or other biotechnological derivatization and enrichment of xanthophyll sources. Suitable forms of lutein are commercially available, such as Floraglo® lutein. The amount of lutein defined herein refers to free lutein (i.e., equivalent to 100% pure lutein).

[0040] The combination according to the invention comprises lutein. In a preferred embodiment, the combination is contained in an oral composition, and preferably lutein is present in an amount of ≥ 5 μg / 100 g dry weight of the composition, preferably 10-1000 μg lutein / 100 g, more preferably 15-750 μg lutein / 100 g, and even more preferably 30-500 μg lutein / 100 g dry weight of the composition.

[0041] 1,3-Dioleoyl-2-palmitoylglycerol (OPO) The composition according to the invention comprises 1,3-dioleoyl-2-palmitoylglycerol (OPO). Preferably, the composition is included in an oral composition, and preferably OPO is present in an amount of 0.5-15 g / 100 g dry weight of the composition, preferably 1-12 g OPO / 100 g, more preferably 2-10 g OPO / 100 g dry weight of the composition.

[0042] Suitable commercially available OPO sources are, for example, from Loders Croklaan, using Betapol. TM The name or Enzymotec Inc.'s InFat TM Alternatively, OPO can be prepared in the manner described, for example, in EP 0698078 and / or EP0758846.

[0043] Indigestible oligosaccharides Preferably, the composition further comprises indigestible oligosaccharides. In a preferred embodiment, the composition is included in an oral composition, and preferably the composition comprises 0.05-25 g of indigestible oligosaccharides per 100 g dry weight of the composition, even more preferably 0.15-15 g of indigestible oligosaccharides per 100 g dry weight of the composition.

[0044] As used in this article, the term “indigestible oligosaccharides” refers to oligosaccharides that are not digested in the gut by digestive enzymes present in the upper digestive tract (e.g., the small intestine and stomach), but instead reach the distal parts of the gut (e.g., the colon) intact, where they are fermented by the gut microbiota.

[0045] Microbial metabolites of indigestible oligosaccharides contain short-chain fatty acids (SCFAs), such as acetate, propionate, butyrate, lactate, etc. The inventors unexpectedly discovered that the presence of fermentation products of indigestible oligosaccharides (i.e., SCFAs) significantly increased lactase expression when used in combination with lutein and 1,3-dioleoyl-2-palmitoylglycerol.

[0046] Preferably, the indigestible oligosaccharide is selected from galactooligosaccharides (GOS), fructooligosaccharides (FOS), human milk oligosaccharides (HMO), or combinations thereof.

[0047] In the embodiments, the indigestible oligosaccharides are preferably selected from FOS, GOS, or combinations thereof. More preferably, the combination and / or oral composition comprises FOS and GOS in a weight ratio of (20 to 2):1, more preferably (16 to 4):1, or even more preferably (12 to 6):1. Most preferably, the weight ratio is about 9:1.

[0048] Preferably, the combination included in the oral composition comprises 0.25-25 g / 100 g dry weight of the composition, more preferably 0.5-10 g / 100 g, and even more preferably 1.5-7.5 g / 100 g dry weight of the composition of FOS, GOS, or combinations thereof.

[0049] GOS is preferably β-galacto-oligosaccharide. β-galacto-oligosaccharide can also be referred to as trans-galacto-oligosaccharide (TOS). GOS is marketed, for example, under the trademarks Vivinal™ (Borculo Domo Ingredients, Netherlands) or Bi2Munno (Classado). Preferably, GOS contains at least 80%, more preferably at least 90%, of β-1,4 and β-1,6 bonds based on total bonds. GOS preferably has a DP of 2-60, more preferably 3-40, even more preferably 4-20, and most preferably 5-10.

[0050] The FOS preferably comprises chains of β-linked fructose units with a DP of 2 to 250, more preferably 5 to 100, and even more preferably 10 to 60. Preferably, the average DP of the FOS is greater than 20. The FOS may preferably contain inulin, levulin, and / or mixed polyfructose. Particularly preferred FOS is inulin. FOS suitable for use in the composition are also commercially available, such as Raftiline® HP (Orafti).

[0051] In the embodiments, the indigestible oligosaccharide is preferably human milk oligosaccharide. "Human milk oligosaccharide" (HMO) is present in human milk and is an indigestible oligosaccharide composed of one or more of the following monomers: D-glucose, D-galactose, N-acetylglucosamine, L-fucose, and sialic acid (N-acetylneuraminic acid).

[0052] The combination included in the oral composition preferably contains 0.05-12 g / 100 g dry weight of the composition, more preferably 0.075-9 g / 100 g, even more preferably 0.15-6 g / 100 g, and most preferably 0.3-3 g / 100 g dry weight of the composition of HMO.

[0053] Preferably, the HMO is selected from sialic oligosaccharides, such as 3'-sialyllactose (3'-SL), 6'-sialyllactose (6'-SL), lactosesialyl-tetrasaccharide a,b,c (LST), disialiallactose-N-tetrasaccharide (DSLNT), sialyl-lactose-N-hexasaccharide (S-LNH), DS-LNH and fucoidan oligosaccharides, such as (un)sulfated fucose oligosaccharides, 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), difucosyllactose, lactose-N-fucopentose (LNFP), I, II, III, IV lactose-N-neofucopentose (LNnFP), lactose-N-difucosyl-hexasaccharide (LNDH) and combinations thereof.

[0054] Preferably, the HMO is selected from 2'-fucosyllactose (2'-FL), 3-fucosyllactose (3-FL), 3-sialyllactose (3'-SL), 6-sialyllactose (6'-SL), lactose-N-tetrasaccharide (LNT), lactose-N-neotetrasaccharide (LNnT), difucosyllactose (DFL), or combinations thereof. More preferably, the HMO is selected from 2'-FL, 3-FL, LNT, 3'-SL, and 6'-SL, or combinations thereof. Most preferably, the HMO is 2'-FL.

[0055] In the embodiments, the indigestible oligosaccharide preferably comprises a combination of GOS and FOS with 2'-FL and / or LNT, and most preferably a combination of GOS, FOS and 2'-FL.

[0056] Preferably, the weight ratio of HMO (preferably 2'-FL) to GOS is 5 to 0.05, more preferably 5 to 0.1, and even more preferably 2 to 0.2. Preferably, the weight ratio of HMO (preferably 2'-FL) to FOS is 10 to 0.05, more preferably 5 to 0.1, and even more preferably 2 to 0.5.

[0057] combination The combination according to the invention comprises lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO). In a preferred embodiment, the combination further comprises an indigestible oligosaccharide selected from galactooligosaccharides (GOS), fructooligosaccharides (FOS), human lactose oligosaccharides (HMO), or combinations thereof, more preferably, the indigestible oligosaccharide selected from galactooligosaccharides and fructooligosaccharides.

[0058] In another preferred embodiment, the combination according to the invention comprises lutein, OPO, and indigestible oligosaccharides, wherein these indigestible oligosaccharides are human milk oligosaccharides selected from 2'-fucosylated lactose (2'-FL), 3-fucosylated lactose (3-FL), 3-sialylated lactose (3'-SL), 6-sialylated lactose (6'-SL), lactose-N-tetrasaccharide (LNT), lactose-N-neotetrasaccharide (LNnT), difucosylated lactose (DFL), or combinations thereof. More preferably, these indigestible oligosaccharides are combinations of GOS and FOS with 2'-FL and / or LNT, and most preferably, combinations of GOS, FOS, and 2'-FL.

[0059] In a preferred embodiment, the combination comprises FOS and GOS in a weight ratio of (20 to 2):1, more preferably (16 to 4):1, or even more preferably (12 to 6):1. Most preferably, the weight ratio is about 9:1. In another preferred embodiment, the combination comprises HMO (preferably 2'-FL) and GOS in a weight ratio of 5 to 0.05, more preferably 5 to 0.1, more preferably 2 to 0.2. Preferably, the combination comprises HMO (preferably 2'-FL) and FOS in a weight ratio of 10 to 0.05, more preferably 5 to 0.1, more preferably 2 to 0.5.

[0060] The use of the combination is as outlined above. Specifically, the combination is used for the prevention or treatment of gastrointestinal disorders in human subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract; preferably, the use is for the treatment of the gastrointestinal disorders. In a preferred embodiment, the use is carried out by oral administration of the combination, preferably by oral administration of a nutritional composition comprising the combination.

[0061] Composition In a preferred embodiment of the invention, the composition is included in an oral composition. The composition is preferably a nutritional composition, more preferably an enteral nutrition composition. Preferably, the nutritional composition is a nutritionally complete composition. The composition preferably comprises digestible carbohydrates, lipids, proteins, and combinations thereof.

[0062] Preferably, the composition comprises digestible carbohydrates. Preferred digestible carbohydrates are lactose, glucose, sucrose, fructose, galactose, maltose, starch, and maltodextrin. Preferably, based on the dry weight of the composition, the composition comprises at least 40 g of digestible carbohydrates per 100 g of the composition, more preferably 45-70 g of digestible carbohydrates per 100 g.

[0063] Preferably, the composition comprises 0-75 g lactose per 100 g dry weight of the composition, more preferably 10-65 g lactose, and most preferably 20-55 g lactose per 100 g dry weight of the composition.

[0064] Preferably, the composition comprises protein. Preferred protein sources are milk protein or plant protein. Preferably, based on the dry weight of the composition, the composition comprises 5-20 g protein / 100 g of the composition, more preferably 8-15 g protein / 100 g.

[0065] Preferably, the composition comprises lipids. Preferred lipid sources are plant lipids, milk lipids, microbial lipids, fish lipids, and combinations thereof. Preferably, based on the dry weight of the composition, the composition comprises 15-35 g / 100 g of the composition, more preferably 20-30 g / 100 g.

[0066] The composition is preferably a powder containing up to 5 wt.% water by weight of the powder. The powder is preferably reconstituted with water or other food-grade aqueous liquid to form a ready-to-drink composition in liquid form. Preferably, 10-15 grams of this powder is added to 90 ml of liquid to obtain 100 ml of ready-to-drink liquid.

[0067] Preferably, the oral composition is infant formula, follow-up formula, or toddler formula. More preferably, the oral composition is infant formula for premature infants and / or small for gestational age infants.

[0068] In this invention, infant formula refers to an artificially manufactured nutritional composition intended for use in infants aged 0 to 4 to 6 months, and intended as a substitute for human milk. Typically, infant formula is suitable as a sole source of nutrition. Such infant formula is also referred to as stage one formula. Follow-up formula, intended for infants aged 4 to 6 months to 12 months, is intended as a supplemental feed for infants beginning weaning and consuming other foods. Infant formula and follow-up formula are subject to strict regulation, such as EU Regulation Nos. 609 / 2013 and 2016 / 127. In the context of this invention, toddler formula refers to an artificially manufactured nutritional composition intended for use in infants aged 12 to 36 months, and intended as a supplemental feed for infants. In the context of this invention, toddler formula may also be referred to as growing milk.

[0069] Preferably, the energy density of infant formula, follow-up formula or toddler formula, when in ready-to-drink form, is 60 kcal to 70 kcal / 100 ml.

[0070] In a preferred embodiment, infant formula, follow-up formula, or toddler formula preferably comprises: -3 to 7 g lipids / 100 kcal, preferably 4 to 6 g lipids / 100 kcal, most preferably 4.5 to 5.5 g lipids / 100 kcal. -1.7 to 3.5 g protein / 100 kcal, more preferably 1.8 to 3.0 g protein / 100 kcal, and most preferably 1.8 to 2.5 g protein / 100 kcal; and -5 to 20 g digestible carbohydrates / 100 kcal, more preferably 6 to 16 g digestible carbohydrates / 100 kcal, and most preferably 10 to 15 g digestible carbohydrates / 100 kcal.

[0071] In a preferred embodiment, the composition comprises digestible carbohydrates, lipids, proteins, and at least 400 kcal of energy per 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 5-1000 μg of lutein; and b. 0.5-15 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0072] In a more preferred embodiment, the composition comprises digestible carbohydrates, lipids, proteins, and at least 400 kcal of energy per 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 30-500 μg of lutein; and b.2-10 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0073] In even more preferred embodiments, the composition comprises digestible carbohydrates, lipids, proteins, and at least 400 kcal of energy per 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 5-1000 μg of lutein; b. 0.25-25 g of indigestible oligosaccharides, comprising fructooligosaccharides and galactooligosaccharides; and c.0.5-15 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0074] In another preferred embodiment, the composition comprises digestible carbohydrates, lipids, proteins, and at least 400 kcal of energy per 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 30-500 μg of lutein; b. 0.5-10 g of indigestible oligosaccharides, including fructooligosaccharides and galactooligosaccharides; and c.2-10 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0075] In another preferred embodiment, the composition comprises digestible carbohydrates, lipids, proteins, and at least 400 kcal of energy per 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following per 100 g: a. 5-1000 μg of lutein; b. 0.25-25 g of indigestible oligosaccharides selected from galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), human milk oligosaccharides (HMOs), or combinations thereof; and c.0.5-15 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0076] Most preferably, the composition comprises digestible carbohydrates, lipids, proteins, and an energy content of at least 400 kcal / 100 g (based on the dry weight of the composition), wherein, based on the dry weight of the composition, the composition further comprises the following / 100 g: a. 30-500 μg of lutein; b. 0.5-10 g of indigestible oligosaccharides, wherein these indigestible oligosaccharides comprise galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), and human milk oligosaccharides (HMO), preferably wherein these indigestible oligosaccharides are combinations of GOS and FOS with 2'-FL and / or LNT, and more preferably combinations of GOS, FOS, and 2'-FL. c.2-10 g 1,3-Dioleoyl-2-palmitoylglycerol.

[0077] In this document and its claims, the verb “comprising” and its inflections are used in their non-limiting sense to mean including the item following the word, but not excluding items not specifically mentioned. Furthermore, reference to an element by the indefinite article “a” or “an” does not preclude the possibility of more than one such element, unless the context explicitly requires the presence of one and only one such element. Therefore, the indefinite article “a” or “an” generally means “at least one” or “one type”.

[0078] The invention is illustrated by the following non-limiting examples.

[0079] Example Materials and Methods Component preparation Concentrated stock solutions were prepared for each test component. Short-chain fatty acids (75% sodium acetate / 20% sodium propionate / 5% sodium butyrate, B5887 / P1880 / S5636, Sigma-Aldrich) were resuspended at a stock solution concentration of 4 M in DMEM medium (31053028, Gibco) and administered at a final concentration of 2 mM. Micelles were used to deliver lutein into cells. Lutein (PHR1699, Sigma-Aldrich) was prepared at a concentration of 1 mg / mL in 310 mM sodium taurocholate and 64.5 mM phosphatidylcholine. The final concentration administered to cells was 0.415 μg / mL lutein. A concentrated stock solution of 2-palmitoylglycerol MAG (OPO) (75614, Sigma-Aldrich) was prepared at 100 mM in ethanol. The final concentration used in the experiment was 100 µM. The final concentration of the component was obtained by dissolving the component stock solution in serum-free DMEM (Gibco™ 31053028) or PBS solution (PBS, 14190144, Gibco; 0.6% BSA; 1.2 mM CaCl2; 1 mM MgCl2).

[0080] Cell culture and maintenance Human Caco-2 cell line (HTB-37 ATCC) was used as a model of intestinal epithelium. Cells (passaged 50-70 times) were maintained in complete growth medium DMEM (high glucose + Glutamax, phenol red, 31966021, Gibraltar), supplemented with 10% heat-inactivated fetal bovine serum (10270106, Gibraltar), 1% penicillin-streptomycin (15140-130, Gibraltar), 1% non-essential amino acids in MEM (MEM NEAA 100X, 11140-035, Gibraltar), and 1% sodium pyruvate (100 mM stock solution, 11360-039, Gibraltar). Cells were grown at 37°C and 5% CO2 and passaged at 80% confluence.

[0081] Caco-2 cells were incubated with the ingredients. Caco-2 cells were loaded at 2.5 × 10⁻⁶. 5Cells were seeded at 1 ml / well in 24-well plates (3526, Corning Costar®, Corning Inc.) and allowed to grow for 21 days. Treatment solutions were prepared by mixing the components in serum-free DMEM (Gibco™ 31053028). The medium in each well was then removed and replaced with the treatment solution (1 ml / well), and incubated at 37°C, 5% CO2 for 16 h. Each treatment was performed in triplicate. A medium control was included as a negative control. After 16 h of incubation, cells were harvested and RNA extracted.

[0082] RNA isolation Following the manufacturer's protocol, total RNA from Caco-2 cells was isolated using TRIzol reagent (15596026, AmbionLife technologies) followed by the RNeasy kit from Qiagen. The quality of the extracted RNA was assessed using a bioanalyzer, with RIN > 8 for each sample (Agilent 2100, SoftMax Pro 7.0 software), and the concentration of each sample was measured using a Nanodrop 2000 spectrophotometer.

[0083] RNA sequencing Paired-end sequence reads were generated using the Illumina NovaSeq 6000 system by BaseClear (Leiden, Netherlands). Sequences generated using NovaSeq 6000 were within the BaseClear BV certification scope (L457; NEN-EN-ISO / IEC 17025). FASTQ read sequence files were generated using bcl2fastq version 2.20 (Illumina). Initial quality assessment was based on data filtered through Illumina Chastity. Subsequently, reads containing PhiX control signals were mapped to the PhiX genome using bowtie2, and aligned reads were removed from the fastq files. Furthermore, reads containing (partial) adapters (minimum read length up to 50 bp) were cleaved using fastq-mcf of the ea-utils package version 1.04. A second quality assessment was based on the remaining reads using the FASTQC quality control tool version 0.11.8.

[0084] The reads were trimmed and quality checked using Trim_Galore version 0.6.2. Next, the RNA-seq reads were mapped to the genomic reference sequence (GCF_000001405.39_GRCh38.p13_genomic) using STAR alignment tool version 2.7.1. Dobin et al., STAR: ultrafast universal RNA-seq aligner, Bioinformatics, 2013, 29(1), 15-21. Gene counts were estimated using featureCounts version 1.6.3 based on the alignment. Liao et al. , featureCounts: An efficient general-purpose program for assigning sequence reads to genomic features. , Bioinformatics , 2014 , 30(7) , (923–930). In addition to the alignment results (BAM file), the results for each sample also provide the raw counts and TPM normalized counts.

[0085] Statistical analysis Differential gene expression analysis was performed using DESeq2 version 1.22.2. Love et al., Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2, Genome Biology, 2014, 15:550. Genes with a q-value (false discovery rate) <0.05 were considered differentially expressed.

[0086] result The results are as follows Figure 1As shown. When compared with the negative control, the following was observed: Lactase expression in Caco-2 cells significantly increased after stimulation with SCFA. The combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) also significantly increased lactase expression. The combination of SCFA, lutein, and 1,3-dioleoyl-2-palmitoylglycerol (OPO) synergistically increased lactase expression in Caco-2 cells.

Claims

1. A combination of lutein and 1,3-dioleoyl-2-palmitoylglycerol (OPO) for use in the prevention or treatment of gastrointestinal disorders in human subjects, wherein the gastrointestinal disorders are caused by impaired expression of lactase in the gastrointestinal tract.

2. The combination for use according to claim 1, wherein the use is for treating the gastrointestinal condition.

3. The combination for use according to claim 1 or 2, wherein the use is carried out by oral administration of the combination, preferably by oral administration of a nutritional composition comprising the combination.

4. The combination for use according to any one of the preceding claims, wherein the treatment of the gastrointestinal symptom is carried out by increasing the expression of lactase in the gastrointestinal tract.

5. The combination for use according to any one of the preceding claims, wherein the combination further comprises an indigestible oligosaccharide selected from galactooligosaccharides (GOS), fructooligosaccharides (FOS), human milk oligosaccharides (HMO), or combinations thereof.

6. The combination for use according to claim 5, wherein the indigestible oligosaccharide is selected from GOS and FOS.

7. The combination for use according to claim 5 or 6, wherein the indigestible oligosaccharide is selected from the HMOs of 2'-fucosylated lactose (2'-FL), 3-fucosylated lactose (3-FL), 3-sialylated lactose (3'-SL), 6-sialylated lactose (6'-SL), lactose-N-tetrasaccharide (LNT), lactose-N-neotetrasaccharide (LNnT), difucosylated lactose (DFL), or combinations thereof.

8. The combination for use according to claim 5, wherein the indigestible oligosaccharide is a combination of GOS and FOS with 2'-FL and / or LNT, preferably a combination of GOS and FOS with 2'-FL.

9. The combination for use according to any one of claims 5-8, wherein the combination is contained in the composition, and wherein the composition contains 0.05-12 g of HMO per 100 g dry weight of the composition.

10. The combination for use according to claim 5, wherein the weight ratio of HMO to GOS is 5 to 0.05, and / or wherein the weight ratio of HMO to FOS is 10 to 0.

05.

11. The combination for use according to any one of the preceding claims, wherein the combination is contained in a composition, the composition further comprising 10-65 g lactose / 100 g dry weight of the composition.

12. The combination for use according to any one of the preceding claims, wherein the impaired lactase expression is due to lactase non-persistence, preterm birth, celiac disease, inflammatory bowel disease, short bowel syndrome, intestinal infection, or exposure of the subject to chemotherapy or a combination thereof.

13. The combination for use according to claim 12, wherein the impaired lactase expression is due to lactase non-persistence, preterm birth, or a combination thereof.

14. The combination for use according to any one of the preceding claims, wherein the symptoms of the gastrointestinal ailment are selected from diarrhea, constipation, flatulence, bloating, abdominal pain, or a combination thereof.

15. The combination for use according to claim 3, wherein the oral administration is administration of infant formula, follow-up formula or toddler formula.

16. The combination for use according to claim 15, wherein the infant formula is for preterm infants and / or small for gestational age infants.

17. The combination for use according to any one of the preceding claims, wherein the combination comprises an oral nutritional composition, and wherein the composition comprises digestible carbohydrates, lipids, proteins, and an energy content of at least 400 kcal / 100 g, based on the dry weight of the composition, wherein, based on the dry weight of the composition, the composition comprises the following / 100 g: a. 30-500 μg of lutein; b. 0.5-10 g of indigestible oligosaccharides, including fructooligosaccharides, galactooligosaccharides, and human milk oligosaccharides; and c.2-10 g 1,3-Dioleoyl-2-palmitoylglycerol.

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