TW201316983A - Composition for use in the promotion of intestinal angiogenesis and of nutrient absorption and of enteral feeding tolerance and/or in the prevention and/or treatment of intestinal inflammation and/or in the recovery after intestinal injury and surgery - Google Patents

Abstract

The invention discloses a composition comprising at least one long chain polyunsaturated fatty acid, at least one probiotic and a mixture of oligosaccharides, said mixture containing at least one N-acetylated oligosaccharide, at least one sialylated oligosaccharide and at least one neutral oligosaccharide, for use in the promotion of intestinal angiogenesis and of nutrient absorption and of enteral feeding tolerance and/or in the prevention and/or treatment of intestinal inflammation, such as necrotizing enterocolitis, and/or in the recovery after intestinal injury and/or surgery. This composition is particularly adapted for use in infants, notably preterm infants.

Description

Composition for promoting intestinal angiogenesis, nutrient absorption and intestinal feeding tolerance, and/or prevention and/or treatment of intestinal inflammation, and/or intestinal damage and post-operative recovery

The present invention relates to for promoting intestinal angiogenesis and nutrient absorption and enteral feeding tolerance, and/or prevention and/or treatment of intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or after surgery Restored composition. This composition is for mammals, preferably humans, and better babies.

The accelerated formation of blood vessels in the intestinal mucosa occurs in the neonatal period, especially in premature babies. Neovascularization and vascular repair are also critical during intestinal injury and/or healing after surgery. Neovascularization and vascular repair occur via angiogenesis.

Infants, in particular premature delivery and LBW infants, can undergo immature intestinal angiogenesis and thus reduce blood flow to the intestinal mucosa (Reber KM, Nankervis CA, Novicki PT. Newborn intestinal circulation. Physiology and pathophysiology. Clin Perinatol 2002; 29 ( 1): 23-39).

Impaired blood flow can delay intestinal development, impair nutrient absorption and tolerance to intestinal feeding, alter intestinal motility, and is believed to be a predisposing factor in the pathogenesis of necrotizing enterocolitis (Nankervis CA, Giannone PJ, Reber KM. The neonatal intestinal vasculature: contributing factors to necrotizing enterocolitis. Semin Perinatol 2008; 32(2): 83-91).

Necrotizing enterocolitis (NEC) primarily affects premature babies. It is the most common emergency surgery for neonatal surgery. About 12% of premature infants weighing less than 1500 g have NEC. Mortality ranges from 20% to 50% and the condition includes stenosis, Adhesive and short bowel syndrome.

A single factor has not yet been determined as the cause of NEC. It is now believed that NEC is the result of a combination of several factors. Two consistent findings were premature birth and feeding. Premature bowel response is abnormal and produces an acute inflammatory response to feeding, leading to intestinal necrosis. Some postnatal problems (including cardiac malformations, intestinal occlusion, infection, or abdominal fissure) are also associated with NEC.

In premature babies, NEC usually occurs between one week and 10 days after the start of feeding. In term infants, NEC occurs within 1 to 4 days of birth if it is raised on the first day. The risk of NEC occurring at a larger gestational age is smaller. Very few NECs occur in unfed infants. Immature neonatal vascular systems are extremely sensitive to environmental stimuli, which can cause significant changes in blood flow. Current findings suggest that mucosal damage caused by vasoconstriction and subsequent ischemia-reperfusion injury may be the initial event in the pathogenesis of NEC. One theory, which associates feeding with intestinal mucosal damage, involves the excessive growth of bacteria when providing a source of enteral nutrients, which causes the bacteria to invade through previously damaged mucosa (Nankervis CA, Giannone PJ, Reber KM. The neonatal intestinal vasculature: contributing Factors to necrotizing enterocolitis. Semin Perinatol 2008;32(2):83-91).

NEC is difficult to prevent. Premature babies who are breastfed or colostrum seem to have a lower incidence of NEC than their formulators (Schanler RJ, Lau C, Hurst NM, Smith EO 2005 Randomized trial of donor human milk versus preterm formula as substitutes for mothers' own Milk in the feeding of extremely premature infants. Pediatrics 116: 400-406). However, it is often not possible to obtain old mother milk or even donor milk.

Medical management includes termination of feeding, nasogastric drainage, antibiotics for 7-14 days, and intravenous nutrition. Fluid state, electrolytes, solidification and oxygen demand must also be closely monitored. 60-80% of NEC infants are medically controlled and the symptoms disappear without surgery. Slowly start feeding after surgery. However, delayed supply of enteral feeding leads to growth stagnation and failure to thrive, which can increase preterm birth morbidity and mortality and lead to sequelae such as impaired cognitive development throughout life.

When medical management is ineffective or intestinal perforation (cracking), surgery is required in 20-40% of cases. The target system only removes the fully necrotic (dead) intestines and leaves any marginal areas in the hope that they will survive. However, surgery leads to short bowel syndrome, which can lead to permanent damage to nutrient digestion, poor tolerance to feeding and impaired quality of life.

The present invention relates to the promotion of intestinal angiogenesis and nutrient absorption and intestinal feeding tolerance, and/or prevention and/or treatment such as necrotizing enterocolitis in infants and young children, preferably infants, in particular. Compositions of intestinal inflammation, and/or intestinal damage and/or recovery after surgery, such infants and young children are born prematurely or have low birth weight (LBW) or undergo intrauterine growth retardation (IUGR) or undergo poor endometrial nutrition And/or intestinal damage and/or surgery.

More generally, the industry needs to perform this nutritional intervention in young mammals, particularly infants and children, preferably infants and young pets.

There is a need in the industry to induce this intervention to maintain or improve the intestinal angiogenesis of humans and animals, especially young mammals.

The inventors have surprisingly found that administration of a particular oligosaccharide mixture with at least one long chain polyunsaturated fatty acid (LC-PUFA) and at least one probiotic combination is particularly effective in promoting intestinal angiogenesis and nutrient absorption and intestinal transit. Feeding tolerance, and/or prevention and/or treatment of intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or post-operative recovery.

Accordingly, the present invention provides a composition comprising at least one LC-PUFA, at least one probiotic, and an oligosaccharide mixture, the mixture comprising at least one N-acetylated oligosaccharide, at least one sialylated oligosaccharide, and at least one Sexual oligosaccharide, which is used to promote nutrient absorption and intestinal angiogenesis and enteral feeding tolerance, and/or to prevent and/or treat intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or Or recover after surgery.

The composition of the invention is preferably a nutritional composition.

The LC-PUFA is preferably selected from the group consisting of arachidonic acid (ARA) and docosahexaenoic acid (DHA), and more preferably, the LC-PUFA is a mixture of ARA and DHA.

The probiotic is preferably selected from a probiotic strain, more preferably, a probiotic strain lactobacillus or bifidobacterium . In a preferred embodiment, the probiotics are Lactobacillus rhamnosus , Bifidobacterium lactis , and Lactobacillus reuteri .

The neutral oligosaccharide is preferably selected from the group consisting of oligofructose (FOS) and galactooligosaccharide (GOS), preferably GOS.

In one embodiment, the oligosaccharide mixture may be derived from animal milk, such as one or more of cow's milk, goat's milk, sheep's milk, or buffalo milk. For example, it is obtained by milk fractionation and further enzymatic treatment.

In a second embodiment, the oligosaccharide mixture can be prepared using enzymatic, chemical enzymatic and/or chemical means.

In a third embodiment, the oligosaccharide mixture can be prepared using yeast and/or bacterial fermentation techniques. For example, yeast and/or bacterial cells that exhibit suitable enzymes (eg, glycosidases and/or glycosyltransferases) can be used for this purpose either after genetic modification or without genetic modification.

The compositions of the present invention are preferably used in infants that are premature or have low birth weight (LBW) or undergo uterine growth retardation (IUGR) and/or poor endometrial nutrition and/or intestinal damage and/or surgery.

The following terms as used herein have the following meanings.

The term "child" means a human being between the stage of birth and the stage of puberty. Adults are older than children.

The term "infant" means a child who is within 12 months of age.

The term "preterm infant or premature infant" means an infant born less than 37 weeks of gestational age.

The term "low birth weight infant" means an infant having a live birth weight of less than 2,500 g.

The term "young child" means a child between the ages of 1 and 3.

The term "infant formula" means a food intended for the specific nutritional use of an infant during the period from 4 months to 6 months prior to life and which satisfies the nutritional requirements of such person (May 14, 1991 regarding infant formula and larger) European Commission Directive of the follow-on formula 91/321/EEC Article 1.2).

The term "premature infant formula" means an infant formula intended for use in a premature infant.

The term "human milk fortifier" means a supplement for calorie, protein, minerals and vitamins in breast milk that is fed to a premature infant or a baby with a low birth weight.

The term "larger infant formula" means a food intended for a particular nutritional use of an infant over 4 months of age and which constitutes a major liquid element in the diet in which such persons are increasingly diversified.

The term "starter infant formula" means a food intended for a particular nutritional use of an infant during the first 4 months of life.

The term "child food" means a food intended for the specific nutritional use of an infant during the first year of life.

The term "baby cereal composition" means a food intended for the particular nutritional use of an infant during the first year of life.

The term "growth milk" means a milk-based beverage that is adapted to the specific nutritional needs of young children.

The term "weaning period" means the period of time during which a portion of a baby's diet partially or completely replaces breast milk or an infant formula.

The term "promoting nutrient absorption and intestinal angiogenesis and enteral feeding tolerance" means supporting nutrient absorption and intestinal angiogenesis and intestinal feeding tolerance. For example, it covers the prevention of ischemia. The term "enteral" means "intragastric".

The term "preventing and/or treating intestinal inflammation" means preventing intestinal inflammation and reducing the frequency and/or incidence and/or severity of intestinal inflammation and/or shortening its Continued time. The incidence refers to the amount of any intestinal inflammation. Frequency refers to the amount of inflammation of the same intestine. This prevention covers reducing the frequency and/or severity of the intestinal inflammation in later life. The term "in later life" covers the effects of the termination of the intervention. The effect of "in later life" may preferably be 2 weeks to 4 weeks, 2 months to 12 months, or 2 to 12 years after the termination of the intervention (eg, 2 years, 5 years, 10 years). Necrotizing enterocolitis is an example of intestinal inflammation.

The term "intestinal injury and/or post-operative recovery" means support for intestinal damage and/or healing and recovery after surgery.

The term "nutritional composition" means a composition that is administered to an individual for nutrition. This nutritional composition is usually administered orally, transgastrically or intravenously, and it typically includes a source of lipid or fat and a source of protein.

The term "synthetic mixture" means a mixture obtained by chemical and/or biological means which is chemically identical to a mixture naturally occurring in mammalian milk.

The term "hypoallergenic composition" means a composition that is unlikely to cause an allergic reaction.

The term "probiotic" means a microbial cell preparation or a microbial cell component or a microbial cell metabolite having a beneficial effect on the health or wellness of the host. (Salminen S, Ouwehand A. Benno Y. et al. " Probiotics: how should they be defined " Trends Food Sci. Technol. 1999: 10 107-10).

The term "oligosaccharide" means a carbohydrate having a degree of polymerization (DP) in the range of 2 to 20 (including 2 and 20), but excluding lactose.

The term "neutral oligosaccharide" means uncharged and has no N-acetyl group residues. Oligosaccharides.

The term "sialylated oligosaccharide" means an oligosaccharide having a residue of a sialic acid (eg, N-ethyl thioneamine and/or N-hydroxyethyl thioglycolic acid).

The term "N-acetylated" oligosaccharide means an oligosaccharide having at least one hexose having an N-acetinyl residue.

All percentages are by weight unless otherwise indicated.

In one aspect, the invention provides a composition comprising: at least one LC-PUFA, at least one probiotic, and an oligosaccharide mixture, the mixture comprising at least one N-acetylation selected from the group consisting of Oligosaccharides: GalNAcα1,3Galβ1,4Glc (=3'GalNAc-lac=N-acetamido-aminogalactosyl-lactose) and Galβ1,6GalNAcα1,3Galβ1,4Glc(=6'Gal-3GalNAc-lac=galactose --N-ethinyl-aminogalactosyl-lactose, Galβ1,4GlcNAcβ1,3Galβ1,4Glc (lacto-N-neotetraose) and Galβ1,3GlcNAcβ1,3Galβ1,4Glc (milk-N-four a sugar); at least one selected from the group consisting of sialylated oligosaccharides comprising: NeuAcα2, 3Galβ1, 4Glc (=3'-sialyl lactose) and NeuAcα2,6Galβ1,4Glc (=6'-sialyl lactose) And at least one group selected from the group consisting of: Galβ1,6Gal (=β1,6-digalactoside), Galβ1,6Galβ1,4Glc (=6'Gal-lac), Galβ1,6Galβ1,6Glc , Galβ1, 3Galβ1, 3Glc, Galβ1, 3Galβ1, 4Glc (=3'Gal-lac), Galβ1,6Galβ1,6Galβ1,4Glc(=6',6-di Gal-lac), Galβ1,6Galβ1,3Galβ1,4Glc(= 6',3-di Gal-lac), Galβ1,3Galβ1, 6Galβ1,4Glc(=3',6-di Gal-lac), Galβ1,3Galβ1,3Galβ1,4Glc(=3',3-di Gal-lac), Galβ1,4Galβ1,4Glc (=4'Gal-lac) and Galβ1,4Galβ1,4Galβ1,4Glc (=4',4-di Gal-lac), and Fucα1,2Galβ1,4Glc (=2'fucosyllactose or FL), which is used to promote nutrient absorption and intestinal blood vessels Neonatal and enteral feeding tolerance, and/or prevention and/or treatment of intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or post-operative recovery.

In a second aspect, the invention relates to a composition comprising at least one long chain polyunsaturated fatty acid, at least one probiotic and an oligosaccharide mixture, the mixture comprising: - 0.25 wt% relative to the total weight of the oligosaccharide mixture At least one N-acetylated oligosaccharide to 20 wt%, preferably 0.3 wt% to 10 wt%, more preferably 0.3 wt% to 5 wt%, and even more preferably about 0.5 wt%, relative to the oligosaccharide At least one sialylated oligosaccharide of from 0.5 wt% to 30 wt%, preferably from 0.75 wt% to 15 wt%, more preferably from 0.75 wt% to 10 wt% and even more preferably from about 1 wt%, based on the total weight of the mixture, And at least one of 50 wt% to 99.3 wt%, preferably 20 wt% to 80 wt%, more preferably 10 wt% to 50 wt%, and even more preferably about 50 wt%, relative to the total weight of the oligosaccharide mixture. Sexual oligosaccharide, which is used to promote intestinal angiogenesis and nutrient absorption and enteral feeding tolerance, and/or prevent and/or treat intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or Or recover after surgery.

According to a preferred embodiment, the oligosaccharide mixture is present in an amount of from 0.5% to 70%, more preferably from 1% to 20%, even more preferably from 2% to 5%, based on the total weight of the composition. in.

The oligosaccharide compound is defined by its structure, wherein GalNAc is N-acetyl galactosamine, GlcNAc is N-acetyl glucosamine, Gal galactose, NeuAc N-acetyl retinoic acid, Fuc system Fucose and Glc are glucose.

The oligosaccharide mixture of the compositions of the invention may be the sole source of oligosaccharides in the composition.

In the first embodiment, the neutral oligosaccharide is preferably selected from the group consisting of FOS and GOS, preferably GOS, such as those described above.

In the second embodiment (independent or not from the first embodiment), the neutral oligosaccharide is preferably 2'-fucosyllactose (FL). In this case, FL is preferably included in the population of neutral oligosaccharides in the oligosaccharide mixture during the manufacture of the oligosaccharide mixture.

Neutral oligosaccharides can be prepared as a mixture by purchasing and mixing the individual components. For example, synthetic galactooligosaccharides such as the following are commercially available under the trade names Vivinal® (from Friesland Campina, Netherlands) and Elix'or®: Galβ1,6Gal, Galβ1,6Galβ1,4Glc, Galβ1,6Galβ1,6Glc, Galβ1,3Galβ1, 3Glc, Galβ1, 3Galβ1, 4Glc, Galβ1, 6Galβ1, 6Galβ1, 4Glc, Galβ1, 6Galβ1, 3Galβ1, 4Glc, Galβ1, 3Galβ1, 6Galβ1, 4Glc, Galβ1, 3Galβ1, 3Galβ1, 4Glc, Galβ1, 4Galβ1, 4Glc and Galβ1,4Galβ1, 4Galβ1, 4Glc and mixtures thereof. Other oligosaccharide suppliers are Dextra Laboratories, Sigma-Aldrich Chemie GmbH and Kyowa Hakko Kogyo Co., Ltd. Alternatively, specific glycosyltransferases and/or glycosidases (eg, galactosyltransferases and/or fucoids may be used) Glycosyltransferases and/or galactosidase and/or fucosidase) to produce galactooligosaccharides and/or fucosylated oligosaccharides.

Fucosyl lactose is a fucosylated oligosaccharide (i.e., an oligosaccharide having a fucose residue). Fucosylated oligosaccharides can be isolated from natural sources (eg, animal milk) by chromatography or filtration techniques. Alternatively, it can be produced by biotechnological means using specific fucosyltransferases and/or fucosidases via the use of enzyme-based fermentation techniques (recombinant or native enzymes) or microbial fermentation techniques. In the latter case, the microorganism can express its natural enzymes and substrates or can be engineered to produce individual receptors and enzymes. Single microbial cultures and/or mixed cultures can be used. The formation of fucosylated oligosaccharides can be initiated by receptor acceptor starting at any degree of polymerization starting from a degree of polymerization (DP)=1. Alternatively, fucosylated oligosaccharides can be produced by chemical synthesis from lactose and free fucose. Fucosylated oligosaccharides are also commercially available, for example, from Kyowa, Hakko, Kogyo (Japan).

According to the present invention, the sialylated oligosaccharide may be selected from the group consisting of 3'-sialyl lactose and 6'-sialyl lactose. Preferably, the sialylated oligosaccharide comprises both 3'-sialyl lactose and 6'-sialyl lactose. In this embodiment, the ratio between the 3'-sialyl lactose and the 6'-sialyl lactose is preferably in the range of between 5:1 and 1:2.

The 3'- and 6'-forms of sialic lactose can be obtained by adding a natural source such as animal milk to the composition, or can be isolated from this natural source by chromatography or filtration techniques. Alternatively, it may be by biotechnological methods using a specific sialyltransferase or sialidase (neuraminidase) by enzyme-based fermentation techniques (recombinant or native enzymes), by chemical synthesis or by microorganism Fermentation technology to produce. In the latter case, the microorganism can express its natural enzymes and substrates or can be engineered to produce individual receptors and enzymes. A single microbial culture or mixed culture can be used. The formation of sialyl oligosaccharides can be initiated by acceptor acceptance starting at any degree of polymerization starting from the degree of polymerization (DP)=1. Alternatively, sialyl lactose can be produced by chemical synthesis from lactose and free N'-ethionylneuraminic acid (sialic acid). Sialyl lactose is also commercially available, for example, from Kyowa Hakko Kogyo (Japan).

N-acetylated oligosaccharides can be obtained by adding a natural source such as animal milk to the composition. Alternatively, it can be prepared by the action of an aglycosidase and/or an aminogalactosidase on N-ethylidene-glucose and/or N-ethylmercaptogalactose. Likewise, N-ethinyl-galactosyltransferase and/or N-ethinyl-glycosyltransferase can be used for this purpose. N-acetylated oligosaccharides can also be produced by fermentation using individual enzymes (recombinant or natural) and/or microbial fermentation using fermentation techniques. In the latter case, the microorganism can express its natural enzymes and substrates or can be engineered to produce individual receptors and enzymes. A single microbial culture or mixed culture can be used. N-acetylated oligosaccharide formation can be initiated by receptor acceptor starting from any degree of polymerization starting from a degree of polymerization (DP)=1. Another option is the chemical conversion of oligosaccharides (such as lactulose) or ketohexoses (such as sugars) bound to the oligosaccharides to N-acetyl hexosamine or N-acetyl hexose hexoses containing oligosaccharides. Amines are described, for example, in Wrodnigg, TM; Stutz, AE (1999) Angew. Chem. Int. Ed. 38: 827-828.

LNnT and LNT can be synthesized by enzymatic transfer of a sugar unit from a donor moiety to a receptor moiety using a glycosyl hydrolase and/or a glycosyltransferase, as described in, for example, U.S. Patent No. 5,288,637 and WO 96/10086. Said. Another option For example, as described in Wrodnigg, TM; Stutz, AE (1999) Angew. Chem. Int. Ed. 38: 827-828, LNnT can be free or bound to an oligosaccharide (eg, lactulose). The ketohexose (for example, fructose) is chemically converted to N-acetyl hexosamine or an oligosaccharide containing N-acetyl hexosamine. The N-ethinyl-lactosamine produced in this manner can then be transferred to the lactose as part of the receptor.

Preferably, the N-acetylated oligosaccharide is selected from the group consisting of milk-N-neotetraose (or LNnT) and milk-N-tetraose (or LNT). Preferably, LNnT and/or LNT are included in the sialylated oligosaccharide in the oligosaccharide mixture during the manufacture of the oligosaccharide mixture.

The probiotic strains present in the compositions of the present invention may be selected from any strain that meets the probiotic definition and has an acceptable shelf life for the composition to be incorporated therein. For example, if the composition is included in an infant formula, the infant formula is required to remain stable and effective for up to 12 months. The probiotic strain is preferably Lactobacillus or Lactobacillus bifidum.

Examples of preferred lactobacilli are Lactobacillus rhamnosus, Lactobacillus paracasei and Lactobacillus reuteri. The preferred strains are Lactobacillus rhamnosus ATCC 53103, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus reuteri DSM 17938 and Lactobacillus paracasei CNCM I-2116. Even better probiotic strains of Lactobacillus rhamnosus, the term encompasses Lactobacillus rhamnosus ATCC 53103 and Lactobacillus rhamnosus CGMCC 1.3724.

Lactobacillus rhamnosus ATCC 53103 is commercially available from Valio Oy (Finland) under the trade name LGG, and Lactobacillus reuteri DSM 17938 is from BioGaia A.B. The trade name Reuteri is for sale.

Examples of preferred Bifidobacterium lactis include Lactobacillus lactis, Bifidobacterium longum , Bifidobacterium breve , and Bifidobacterium infantis , especially strains. Lactobacillus acidophilus CNCM I-3446 sold by Christian Hansen (Denmark) under the trade name Bb12, Lactobacillus longiflorum ATCC BAA-999 sold by Morinaga Milk Industry Co., Ltd. (Japan) under the trade name BB536, by Danisco Lactobacillus brevis strains sold under the trade name Bb-03, Lactobacillus brevis strains sold under the trade name M-16V by Morinaga, Lactobacillus bifidus strains sold under the trade name Bifantis by Procter & Gamble and A strain of Lactobacillus brevis that sold by Institut Rosell (Lallemand) under the trade name R0070.

According to the present invention, the probiotic strain is selected from the group of probiotic bacteria, and the probiotic bacteria are preferably Lactobacillus or Lactobacillus bifidum, and the probiotics are more preferably Lactobacillus rhamnosus, Lactobacillus reuteri and Lactobacillus lactici.

Probiotics can be present in the composition in a wide range of percentages, provided that the probiotics deliver the effect. Preferably, however, the probiotic strain is present in a combination of a probiotic strain equivalent to 10e2 cfu (=colony forming unit) to 10e12 cfu per gram of composition, more preferably between 10e6 cfu and 10e9 cfu. In. This expression includes the possibility that the bacteria are viable, inactive or dead or even present as fragments (eg DNA, cell wall material, intracellular material or bacterial metabolite). In other words, if all bacteria are alive (whether they are actually alive, inactive or dead, fragmented or otherwise) The amount of bacteria contained in the composition, expressed as the bacterial colony forming ability of the amount of bacteria.

The composition contains at least one LC-PUFA, which is typically an n-3 or n6 LC-PUFA. The n-3 LC-PUFA can be a C20 or C22 n-3 fatty acid. Preferably, the C20 or C22 n-3 LC-PUFA is present in an amount of at least 0.1 wt% of all fatty acids in the composition. Preferably, n-3 LC-PUFA is docosahexaenoic acid (DHA, C22: 6, n-3). The n-6 LC-PUFA can be a C20 or C22 n-6 fatty acid. Preferably, the C20 or C22 n-6 LC-PUFA is present in an amount of at least 0.1 wt% of all fatty acids in the composition. Preferably, n-6 LC-PUFA is arachidonic acid (ARA, C20: 4, n-6). The source of LC-PUFA can be, for example, egg fat, fungal oil, low EPA fish oil or algae oil. The LC-PUFA of the composition of the present invention can be provided in a small amount of oil (e.g., fish oil or microbial oil) containing a large amount of pre-formed arachidonic acid and docosahexaenoic acid.

The composition of the present invention is preferably a nutritional composition, more preferably a synthetic nutritional composition. In this case, it may be a premature infant formula, a human milk fortifier, a neonatal formula, a larger infant formula, a toddler food formula, a baby cereal formula, a growing milk, a medical food for clinical nutrition, or a hospitalization usually Supplements used during and/or after discharge from the hospital. Supplements can be used for premature babies or children or adults. Preferably, the composition is for use in preterm delivery of a product, such as a premature infant formula, a human milk fortifier or a premature infant supplement. According to one embodiment, the composition is preferably a premature infant formula, a human milk fortifier or a supplement. The composition of the invention may also be a product for children or adults (such as yogurt or medical food) as well as pet food.

According to a particularly preferred embodiment, the composition of the invention is for preterm labor or has LBW Or infants and young children experiencing IUGR are preferred for infants who experience IUGR and/or experience adverse intrauterine nutrition and/or intestinal damage and/or surgery.

The compositions of the invention may be used before and/or during and/or after the weaning period.

The invention also encompasses the use of a composition of the invention as a synthetic nutrient for promoting intestinal angiogenesis and nutrient absorption and intestinal feeding tolerance, and/or prevention and/or treatment of intestinal inflammation (such as necrotizing enterocolitis) ) and/or intestinal damage and/or recovery after surgery.

In the case of humans, all of the uses described above are particularly intended for use in infants and young children, preferably infants. However, such uses are also intended for younger pets. The compositions and uses of the present invention are particularly suitable for infants and young children (preferably infants) who are premature or have LBW or who experience IUGR and/or poor endometriosis and/or intestinal damage and/or surgery.

Without wishing to be bound by theory, the inventors believe that combinations of oligosaccharide mixtures in the above compositions are effective in intestinal angiogenesis and nutrient absorption and enteral feeding tolerance, and/or prevention and/or treatment such as necrotizing small intestine The efficacy of inflammation such as inflammation of the intestines, and/or intestinal damage and/or post-operative recovery may be the result of a synergistic combination of pro-bacterial strains and LC-PUFAs that are triggered by stimulation with a particular oligosaccharide mixture.

The oligosaccharide mixture, LC-PUFA and probiotic strain can be administered in the same composition or can be administered sequentially.

If it is desired to cope with the preterm and LBW infants, the composition is preferably a nutritional composition that is consumed, for example, in liquid form. It can be a nutritionally complete formula such as (premature) infant formula, supplements, human milk fortifiers, larger infant formulas or Growing milk. Alternatively, for young mammalian groups, the composition can be a pet food.

The compositions of the invention may also contain a source of protein. It is believed that the type of protein is not critical to the invention, provided that the minimum requirements for the essential amino acid are met and satisfactory growth is ensured. Thus, protein sources based on whey, casein, and mixtures thereof, as well as soy-based protein sources can be used. In the case of whey proteins, the protein source may be based on acid whey or sweet whey or mixtures thereof and may include alpha-lactalbumin and beta-lactoglobulin in any desired ratio. The protein can eventually be partially hydrolyzed to enhance oral tolerance to allergens, especially food allergens. In this case, the composition is a hypoallergenic composition.

In addition to the oligosaccharide mixture, the compositions of the invention may also contain a source of carbohydrates. This is especially preferred in the case where the composition of the invention is an infant formula. In this case, although any carbohydrate source commonly found in infant formulas, such as lactose, sucrose, maltodextrin, starch, and mixtures thereof, may be used, the preferred carbohydrate source is lactose. In either case, the oligosaccharide mixture is preferably a single source of prebiotics in the compositions of the invention.

In addition to LC-PUFA, the compositions of the invention may also contain a source of lipids. This is especially suitable if the nutritional composition of the invention is an infant formula. In this case, the lipid source can be any lipid or fat suitable for use in an infant formula. Preferred fat sources include palmitoleic acid, high oleic sunflower oil, and high oleic safflower oil. The essential fatty acids linoleic acid and alpha-linolenic acid can also be added. In the composition, the fat source (including LC-PUFA, such as ARA and/or DHA) preferably has an n-6 to n of from about 1:2 to about 10:1, preferably from about 3:1 to about 8:1. -3 fat The ratio of acid.

The compositions of the present invention may also contain all of the vitamins and minerals deemed necessary in the daily diet in a nutritionally significant amount. Minimum requirements for certain vitamins and minerals have been established. Examples of minerals, vitamins and other nutrients which are optionally present in the compositions of the present invention include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, Nicotinic acid, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine and L-carnitine. Minerals are usually added in the form of a salt. The presence and amount of specific minerals and other vitamins will vary depending on the intended population.

If desired, the compositions of the present invention may contain emulsifiers and stabilizers such as soy lecithin, citric acid esters of mono- and diglycerides, and the like.

The compositions of the present invention may also contain other substances which may have beneficial effects, such as lactoferrin, nucleotides, nucleosides, gangliosides, polyamines, and the like.

The preparation of the compositions of the invention will now be illustrated by way of example.

The formulation can be prepared in any suitable manner. For example, it can be prepared by blending a protein source, a carbohydrate source (as opposed to an oligosaccharide mixture) and a fat source (including LC-PUFA) in an appropriate ratio. If used, an emulsifier can be included at this time. Vitamins and minerals can be added at this point, but they are usually added later to avoid thermal degradation. Any lipophilic vitamins, emulsifiers, and the like can be dissolved in the fat source prior to blending. Water that has preferably been subjected to reverse osmosis can then be mixed to form a liquid mixture. Water temperature is convenient It is in the range of between about 50 ° C and about 80 ° C to aid in the dispersion of the ingredients. Commercially available liquefiers can be used to form the liquid mixture. If the final product is to be in liquid form, the oligosaccharide mixture will be added at this stage. If the final product is to be a powder, the oligosaccharides may be added at this stage if desired. The liquid mixture is then homogenized in, for example, two stages.

The liquid mixture can then be heat treated by, for example, rapidly heating the liquid mixture to a temperature in the range between about 80 ° C and about 150 ° C and maintaining a duration between about 5 seconds and about 5 minutes. To reduce the bacterial load. This can be carried out by means of a steam injection, an autoclave or a heat exchanger (for example a plate heat exchanger).

The liquid mixture can then be cooled to between about 60 ° C and about 85 ° C by, for example, flash cooling. The liquid mixture can then be homogenized again, for example, in two stages, between about 10 Mpa and about 30 Mpa in the first stage, and between about 2 Mpa and about 10 Mpa in the second stage. . The homogenized mixture can then be further cooled to add any heat sensitive components such as vitamins and minerals. At this time, the pH and solid content of the homogenized mixture can be conveniently adjusted.

The homogenized mixture is transferred to a suitable drying device (eg, a spray dryer or freeze dryer) and converted to a powder. The moisture content of the powder should be less than about 5% by weight. The oligosaccharide mixture can be added at this stage by dry mixing with the probiotic strain or by blending it with the probiotic strain in the form of a crystalline syrup, and spray dried (or freeze dried).

If the liquid composition is preferred, the homogenized mixture can be sterilized and then aseptically filled into a suitable container, or it can be first filled into a container and then passed through Sterilization tank treatment.

In another embodiment, the compositions of the invention may be in an amount sufficient to achieve the desired effect in the individual. This form of administration is generally more suitable for preterm or LBW or IUGR infants, older children and adults.

The amount of the oligosaccharide mixture, LC-PUFA and probiotic strain to be included in the supplement will be selected according to the manner in which the supplement is to be administered.

The supplement may be in the form of, for example, a powder, a lozenge, a capsule, a soft lozenge or a liquid. Supplements may further contain protective hydrocolloids (eg, gums, proteins, modified starches), binders, film formers, encapsulants/materials, wall/shell materials, matrix compounds, coatings, emulsifiers, surfactants , solubilizer (oil, fat, wax, lecithin, etc.), adsorbent, carrier, filler, co-compound, dispersant, wetting agent, processing aid (solvent), flow agent, taste mask, weight gain Agent, gelling agent and gel former. Supplements may also contain customary pharmaceutical additives and adjuvants, excipients and diluents including, but not limited to, water, gelatin of any origin, vegetable gums, sulfonic acid lignin, talc, sugar, starch, gum arabic ( Gum arabic), vegetable oils, polyalkylene glycols, flavoring agents, preservatives, stabilizers, emulsifiers, buffers, lubricants, colorants, wetting agents, fillers, and the like.

The supplement may be added to a product acceptable to the consumer (which is a human or animal) (for example, an ingestible carrier or carrier, respectively). Examples of such carriers or carriers are pharmaceutical or food or pet food compositions. Non-limiting examples of such compositions are milk, yogurt, curd, cheese, fermented milk, milk based fermentation products, fermented cereal based products, milk based powders, human milk, preterm formula, infant formula, oral supplements And tube feeding.

In addition, supplements may contain organic or inorganic carrier materials suitable for enteral or parenteral administration, as well as vitamins, trace mineral elements, and other micronutrients, as recommended by government agencies such as USRDA.

Advantages, properties, and various other features of the present invention will become more fully apparent from the <RTIgt;

Instance

Experiments were carried out on the addition of an oligosaccharide mixture (which is a mixture of galactooligosaccharide-rich bovine milk oligosaccharide (CMOS)) (demineralized lactose whey permeate or DDWP), LC-PUFA (arachidonic acid) Effects of -ARA- and docosahexaenoic acid-DHA-) and Lactobacillus lactis (BL) and optionally 2'-fucosyllactose (FL) on juveniles.

Method Experimental program

The experiments were performed in accordance with the Swiss Animal Protection Law and approved by Office Vétérinaire Cantonal (Lausanne, Switzerland, Grant Nos. 2028 and 2028.1). Reproductor male and unmarried female Long-Evans Hooded rat lines were purchased from Janvier (France) and the rats arrived at the animal care facility two weeks prior to mating.

Pregnant females receive food (Kliba 3437) and water ad libitum, kept in constant temperature and humidity, and maintained at 12:12 dark: bright cycle. Captive conditions maintain the full duration of the program. On the second day after birth (B) after birth (B), the mother is removed from the maternity cage and the sex of the cub is determined. After randomization of body weight, 8 standardized litters of male pups were allocated for rearing. Assigning two breeding conditions to the mother and her cubs One of the following: 1) maternal deprivation group, which is isolated from the mother for 180 min (MS) per day from PND2 to PND14, or 2) untreated control (NS).

MS pups were weaned (W) at PND15. They were randomly assigned by weight and lactating mothers and distributed to a group of 16 animals, which received the following feeding until PND26: control diet (modified AIN 93G, MS control group) or suitable for LC-PUFA, lactate double A similar diet of Lactobacillus fermentum CNCM I-3446 (BL) and oligosaccharide (DDWP). MS animals (MS control group) were housed in groups of 8 pups up to PND21 to reduce the stress load of premature weaning, and then individually housed until the end of the experiment. Animals in the NS group were weaned to a control diet (NS control group) at PND21 and individually housed until the end of the experiment.

Animals were sacrificed (+) in PND26 by thorough bleeding under isoflurane anesthesia. Intestinal samples were collected for further qRT-PCR analysis of the expression of angiogenic marker mRNA.

2. Treatment and diet

The following functional ingredients for the experimental tube feeding and dietary compositions contained 98.8% dry matter DDWP ingredients, the composition of which is detailed in Table 1 below.

DDWP is generally obtained according to the disclosure of WO 2007/101675 or WO 2007/090894 and usually contains a mixture of about 30 wt% of GalNAcα1,3Galβ1,4Glc and Galβ1,6GalNAcα1,3Galβ1,4Glc; 50 wt% of Galβ1,6Galβ1 4Glc and Galβ1, 3Galβ1, 4Glc; 20 wt% of NeuAcα2, 3Galβ1, 4Glc and NeuAcα2, 6Galβ1, 4Glc.

Animals were weaned from the time of weaning until the end of the experiment: a semi-synthetic diet for nutritional adaptation (modified AIN 93 G), the composition of which is shown in Tables 2, 3 and 4.

Kjeldhal analysis; ⁶ by Soxhlet; ⁷ AA = arachidonic acid, ⁸ DHA = docosahexaenoic acid; ⁹ by standard culture method and PCR analysis; NA = not analyzed; ND = below detection limit ( Less than 1.00E + 03)

The fatty acid profiles of the three diets were balanced to provide similar ratios of n-6/n-3 and similar ratios of saturated, monounsaturated and polyunsaturated fatty acids. Therefore, the fatty acid composition of the three diets is nearly identical in terms of fatty acid profile.

Animals were sacrificed on the 26th day after birth (PND).

3. Angiogenesis parameters

A marker for angiogenesis, namely CD34, which is a highly expressed protein in endothelial cells, and VEGF (vascular endothelial growth factor) and FGF (fibroblast growth factor), which are angiogenic factors, are used.

It is known that the basic principles of all free indicators CD34, VEGF and FGF are indicative of better vascular formation in the intestine.

Based on the results of Figures 2, 3 and 4, it appears that the compositions of the invention (PUFA-BL-DDWP and PUFA-BL-DDWP-FL) exhibit an angiogenic effect of angiogenic growth factors and markers over both compositions. (Control NS control and MS control). In particular, normalized mRNA expression of CD34 and VEGF was even better than the control NS control.

Thus, the inventive mixtures (PUFA-BL-DDWP and PUFA-BL-DDWP-FL) demonstrate that the vascularization of the present invention is superior to the MS-control and is also superior to the NS-control. This is a true advantage of the compositions of the invention.

It is known that impaired blood flow delays intestinal development, thereby impairing nutrient absorption, and we believe it is a predisposing factor in the pathogenesis of NEC. Therefore, the mixture (PUFA-BL-DDWP and PUFA-BL-DDWP-FL) effectively helps to ingest one of them. The body does not experience a decrease in blood flow to the intestinal mucosa.

Thus, the nutritional composition of the present invention is effective in intestinal angiogenesis and nutrient absorption and enteral feeding tolerance, and/or prevention and/or treatment of intestinal inflammation such as necrotizing enterocolitis, and/or intestinal damage and/or surgery The post-recovery aspect shows the effect.

Figure 1 is a diagram illustrating the experimental protocol.

Figure 2 is a bar graph plotting experimental results for normalized mRNA expression of CD34 (x 10 ^-3 ).

Figure 3 is a bar graph plotting experimental results for normalized mRNA expression of VEGF (x 10 ^-3 ).

Figure 4 is a bar graph showing experimental results of normalized mRNA expression of FGF (x 10 ^-3 ).

Claims (13)

A composition comprising at least one long chain polyunsaturated fatty acid (LC-PUFA), at least one probiotic and an oligosaccharide mixture, the mixture comprising at least one N-acetylated oligosaccharide, at least one sialylated oligosaccharide And at least one neutral oligosaccharide for promoting intestinal angiogenesis and nutrient absorption and enteral feeding tolerance, and/or preventing and/or treating intestinal inflammation (such as necrotizing enterocolitis), and/or Or intestinal damage and / or recovery after surgery. The composition of claim 1, wherein the neutral oligosaccharide is selected from the group consisting of fructooligosaccharides (FOS) and/or galactooligosaccharides (GOS), preferably GOS. The composition of claim 1 or 2, wherein the oligosaccharide mixture contains at least one N-acetylated oligosaccharide selected from the group consisting of GalNAcα1, 3Galβ1, 4Glc (=3'GalNAc-lac=N-B Mercapto-aminogalactosyl-lactose, Galβ1,6GalNAcα1,3Galβ1,4Glc (=6'Gal-3GalNAc-lac=galactosyl-N-ethylidene-aminogalactosyl-lactose), Galβ1, 4GlcNAcβ1, 3Galβ1, 4Glc (lacto-N-neotetraose or LNnT) and Galβ1, 3GlcNAcβ1, 3Galβ1, 4Glc (milk-N-tetraose or LNT); at least one selected from the group consisting of the following sialyl groups Oligosaccharides: NeuAcα2, 3Galβ1, 4Glc (=3'-sialyl lactose) and NeuAcα2,6Galβ1,4Glc (=6'-sialyl lactose); and at least one group of neutral oligosaccharides selected from the group consisting of :Galβ1,6Gal(=β1,6-digalactoside), Galβ1,6Galβ1,4Glc(=6'Gal-lac), Galβ1,6Galβ1,6Glc, Galβ1,3Galβ1,3Glc, Galβ1,3Galβ1,4Glc(=3 'Gal-lac', Galβ1, 6Galβ1, 6Galβ1, 4Glc (=6',6-di-Gal-lac), Galβ1,6Galβ1, 3Galβ1,4Glc(=6',3-di Gal-lac), Galβ1,3Galβ1,6Galβ1,4Glc(=3',6-di Gal-lac), Galβ1,3Galβ1,3Galβ1,4Glc(=3',3- Two Gal-lac), Galβ1, 4Galβ1, 4Glc (=4'Gal-lac) and Galβ1,4Galβ1,4Galβ1,4Glc (=4',4-di Gal-lac), and Fucα1,2Galβ1,4Glc (=2' Fucosyl lactose or FL). The composition of claim 1 or 2, wherein the oligosaccharide mixture comprises: 0.25 wt% to 20 wt%, preferably 0.3 wt% to 10 wt%, more preferably 0.3 wt%, based on the total weight of the oligosaccharide mixture Up to 5 wt% and even more preferably about 0.5 wt% of at least one N-acetylated oligosaccharide, from 0.5 wt% to 30 wt%, preferably from 0.75 wt% to 15 wt%, based on the total weight of the oligosaccharide mixture %, more preferably 0.75 wt% to 10 wt% and even more preferably about 1 wt% of at least one sialylated oligosaccharide, and 50 wt% to 99.3 wt%, preferably relative to the total weight of the oligosaccharide mixture. 20 wt% to 80 wt%, more preferably 10 wt% to 50 wt%, and even more preferably about 50 wt% of at least one neutral oligosaccharide. The composition of claim 1 or 2, wherein the oligosaccharide mixture is present in an amount of from 0.5% to 70%, more preferably from 1% to 20%, even more preferably from 2% to 5%, based on the total weight of the composition. . The composition of claim 1 or 2, wherein the LC-PUFA is selected from the group consisting of arachidonic acid (ARA) and docosahexaenoic acid (DHA), and the LC-PUFA is preferably a mixture of ARA and DHA. The composition of claim 1 or 2, wherein the probiotic strain is selected from the group of probiotic bacteria, preferably a lactobacillus or a bifidobacterium , the probiotic being better than rhamnose Lactobacillus rhamnosus , Bifidobacterium lactis and Lactobacillus reuteri . The composition of claim 1 or 2, wherein the N-acetylated oligosaccharide is selected from the group consisting of milk-N-neotetraose (or LNnT) and milk-N-tetraose (or LNT). The composition of claim 1 or 2, wherein the sialylated oligosaccharide is selected from the group consisting of 3'-sialyl lactose and 6'-sialyl lactose, and the sialylated oligosaccharide is preferably Containing both 3'-sialyl lactose and 6'-sialyl lactose, the ratio between 3'-sialyl lactose and 6'-sialyl lactose is preferably between 5:1 and 1:2 Within the range. The composition of claim 1 or 2, wherein the neutral oligosaccharide is 2'-fucosyllactose (or FL). The composition of claim 1 or 2, wherein the composition is a premature infant formula, a human milk fortifier, a starter infant formula, a follow-on formula, a toddler food formula, a baby cereal Formulation, growing milk, medical food or supplement for clinical nutrition, and the composition is preferably a premature infant formula, a human milk fortifier or a supplement. A composition according to claim 1 or 2 for use in infants and children having premature birth or having low birth weight or experiencing intrauterine growth retardation and/or undergoing suboptimal intrauterine nutrition and/or intestinal damage and/or surgery Preferably, it is an infant. Use of a composition comprising at least one long chain polyunsaturated fatty acid (LC-PUFA), at least one probiotic and an oligosaccharide mixture, the mixture Containing at least one N-acetylated oligosaccharide, at least one sialylated oligosaccharide, and at least one neutral oligosaccharide, the composition being used for the manufacture of a synthetic nutrient for promoting intestinal angiogenesis and nutrient absorption and Intestinal feeding tolerance, and/or prevention and/or treatment of intestinal inflammation (such as necrotizing enterocolitis), and/or intestinal damage and/or post-operative recovery.