TW201316982A - Composition for use in the promotion of healthy bone growth and/or in the prevention and/or treatment of bone disease - 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 healthy bone growth and/or in the prevention and/or treatment of bone disease. Said bone disease is in particular osteomalacia, rickets, osteopenia or osteoporosis. Preferably the composition is a nutritional composition.

Description

Composition for promoting healthy bone growth and/or preventing and/or treating bone diseases

The present invention relates to a composition for preventing and/or treating bone diseases (for example, promoting healthy bone growth) and for preventing or treating bone mass loss, osteomalacia or rickets or osteoporosis. This composition is for use in mammals, preferably humans.

The reduction in bone mass is characterized by a lack of organic bone matrix resulting in a lower than normal condition of bone tissue.

Bone softening disease has lower bone mineral density (BMD) and bone mineral content (BMC) than normal conditions.

Both conditions occur frequently in premature and/or low birth weight infants and/or infants suffering from poor uterine nutrition and result in increased risk of fracture in these groups (Arch Dis Child Fetal Neonatal Ed 2002 86: F82-F85 ). These conditions also frequently affect infants, children and adolescents who have experienced growth retardation due to malnutrition and/or disease.

Children with bony softening are usually associated with rickets, and bone loss is often limited to wilder (adult form of disease). Many doctors often believe that osteomalacia is a precursor to osteoporosis.

Osteoporosis ("porous bone", from Greek) is a bone disease that causes an increased risk of fracture. This disease is characterized by too little bone formation, excessive bone loss, or a combination of both. In osteoporosis, bone mineral density (BMD) is reduced, bone microstructure is degraded, and the amount and type of protein in the bone changes.

Osteoporosis is defined by the World Health Organization (WHO) as having a bone mineral density (BMD) below the mean peak bone mass (average of healthy young adults) of 2.5 standard deviations or greater, as by DXA ( Double energy X-ray absorptiometry). For comparison, bone mass reduction is defined as the standard deviation of bone mineral density below the average peak bone mass between 1.0 and 2.5. The BMD test provides the individual with a measure called the T-score, which is the value obtained by comparing the individual's bone density with the optimal bone density.

The osteomalacia system has a general bone condition in which bone mineralization is insufficient. Osteopathic softening or rickets can show signs of diffuse body pain, muscle weakness, and fragility of the bones. The most common cause of the disease is the lack of vitamin D, which is usually obtained from diet and sun exposure.

However, there is still a need in the industry for nutritional compositions for the prevention and/or treatment of bone diseases (especially osteopenia, osteodystrophy, rickets and osteoporosis), particularly for infants and young children, for premature delivery of such infants and young children. Or have low birth weight (LBW) or experience intrauterine growth retardation (IUGR), or they often experience growth retardation or experience such as Crohn's disease and/or due to malnutrition (usually due to poor endometrial nutrition) Or diseases such as celiac disease and/or cancer, or treatment with drugs (eg, chemotherapeutic drugs and/or corticosteroids) resulting in malabsorption, anorexia, and/or metabolic bone disease.

The industry needs to improve bone diseases such as osteopenia, osteodystrophy, rickets or osteoporosis by being compatible with weak individuals such as infants or children and not based on drug-based interventions.

The industry needs to reduce the frequency, incidence, severity and/or severity of bone diseases. Or shorten the long-term effect of its duration. In addition, the industry needs to be “measured in the future”, especially after years of intervention.

There is a need in the industry for nutritional interventions in young mammals (particularly infants and children, as well as young pets) that are prematurely born or have LBW or undergo IUGR, or that suffer from growth retardation or experience such as Crohn's due to malnutrition Diseases and/or diseases such as celiac disease and/or cancer, or treatment with drugs such as chemotherapeutic drugs and/or corticosteroids, leading to malabsorption, anorexia and/or metabolic bone disease.

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

The inventors have surprisingly discovered 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 healthy bone growth and preventing and/or treating bones. Diseases, in particular osteomalosis, rickets, osteopenia and osteoporosis.

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 A sexual oligosaccharide which is useful for the prevention and/or treatment of bone diseases, preferably osteodystrophy, rickets, osteopenia or osteoporosis.

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 probiotic strain Lactobacillus rhamnosus (the term encompasses Lactobacillus rhamnosus ATCC 53103 and Lactobacillus rhamnosus CGMCC 1.3724, aliased NCC4007 and LPR), Royth lactic acid Lactobacillus reuteri and Bifidobacterium lactis .

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 invention encompasses the use of three different types of compositions of the invention. In the first case, the individual, and in particular the child, is healthy and does not have any risk of bone disease because of a history of no bone disease in the family. In the second case, the individual and, in particular, the child are healthy, but have a risk of bone disease because of a history of bone disease in their family or because of their premature birth or LBW or experiencing IUGR, or they are experiencing growth retardation Time period or experience such as gram Diseases such as Long's disease and/or celiac disease and/or cancer, or drugs that cause malabsorption, anorexia and/or metabolic bone disease (eg, chemotherapeutic drugs and/or corticosteroids). In the third case, the individual, and in particular the child, has a bone disease and is therefore a patient. A preferred use of the invention is the second case for preterm/LBW/IUGR infants and also for infants/children who are subjected to growth delays due to disease or malnutrition or drug use.

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 "premature baby" 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 calorie, protein used to increase the amount of milk fed to premature babies or infants with low birth weight. A supplement of quality, minerals and vitamins.

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 "skeletal disease" refers to a medical condition affecting bone, specifically a condition associated with a decrease in bone organic matrix (eg, reduced bone mass or osteoporosis) and/or a condition associated with reduced bone mineralization (eg osteomalacia) And rickets).

The term "preventing and/or treating a bone disorder" means preventing the bone disease and reducing the frequency and/or incidence and/or severity of the bone disease and/or shortening its duration. Incidence is the number of any bone disease. Frequency refers to the number of diseases of the same bone. This prevention covers reducing the frequency and/or severity of the bone disease in later life. The term "in later life" covers the effects of the termination of the intervention. The effect of "in the future life" Preferably, it may be 2 weeks to 4 weeks, 2 months to 12 months, or 2 to 12 years after the termination of the intervention (for example, 2 years, 5 years, 10 years). The fracture can be the result of a bone disorder, such as those described above.

The term "promoting healthy bone growth" means supporting bone growth, so it is achieved in the most balanced way.

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 usually includes a lipid or fat source and a protein source.

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 microbial cell component that has 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 an oligosaccharide that is uncharged and does not have an N-acetinyl residue.

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 having at least one N-acetamidine An oligosaccharide of a hexose of a residue.

NCC refers to the Nestlé Culture Colle- tion.

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 a 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), Galβ1,6GalNAcα1,3Galβ1,4Glc (=6'Gal-3GalNAc-lac=galactose --N-ethinyl-aminogalactosyl-lactose), Galβ1,4GlcNAcβ1,3Galβ1,4Glc (lacto-N-neotetraose or LNnT) and Galβ1,3GlcNAcβ1,3Gal β1,4Glc (milk- N-tetraose or LNT); at least one selected from the group consisting of sialylated oligosaccharides: NeuAcα2, 3Galβ1, 4Glc (=3'-sialyl lactose) and NeuAcα2,6Galβ1,4Glc (=6'- a sialyl lactose; and at least one group of intermediate oligosaccharides selected from the group consisting of Galβ1,6Gal (=β1,6-digalactoside), Galβ1,6Galβ1,4Glc (=6'Gal-lac), Galβ1,6Galβ1,6Glc,Galβ1,3Galβ1,3Glc,Galβ1,3Gal β1,4Glc(=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), The composition is for promoting healthy bone growth and preventing and/or treating bone diseases, preferably osteodynacia, rickets, osteopenia or osteoporosis.

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 mixture a total weight 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%, of at least one sialylated oligosaccharide, and At least one neutrality of from 50 wt% to 99.3 wt%, preferably from 20 wt% to 80 wt%, more preferably from 10 wt% to 50 wt%, and even more preferably from about 50 wt%, relative to the total weight of the oligosaccharide mixture. Oligosaccharide, which is used to promote healthy bone growth and to prevent and/or treat bone diseases, preferably osteomosis, rickets, osteopenia or osteoporosis.

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%, relative to the total weight of the composition.

The oligosaccharide compound is defined by its structure, wherein GalNAc is N-acetyl galactosamine, GlcNAc is N-acetyl glucosamine, Gal galactose, NeuAc N-ethyl thione, Glc Glucose and Fuc are fucose.

The oligosaccharide mixture of the compositions of the invention is preferably 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, 3GaIβ1, 3Galβ1, 4Glc, Galβ1, 4Galβ1, 4Glc and Galβ1,4GaIβ1, 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 (such as galactosyltransferase and/or fucosyltransferase and/or galactosidase and/or fucosidase) can be used to generate Semi-milk Sugar 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 Microbial fermentation technology is produced. In the latter case, the microorganism can express its natural enzyme And the substrate may be modified to produce a separate substrate and enzyme. 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 is as in Wrodnigg, T.M.; Stutz, A.E. (1999) Angew.Chem. As described in Int. Ed. 38:827-828, LNnT can be chemically converted to N-ethylhexyl hexose by ketohexose (eg, fructose) which is free or bound to an oligosaccharide (eg, lactulose) It is prepared by an amine or an oligosaccharide containing N-ethionyl 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. Lactobacillus reuteri DSM 17938 is commercially available from BioGaia A.B under the trade name Reuteri.

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 an embodiment of 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 better 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, inactivated or dead, fragmented, or a mixture of any or all of these states), the amount of bacteria contained in the composition Take The colony forming ability of this amount of bacteria is expressed.

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 usually during hospitalization. Supplements for use and/or intended to be used after discharge. 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 an 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 use in infants and young children who are born prematurely or have LBW or undergo IUGR, preferably for infants: Experience IUGR, or it usually suffers from growth retardation due to malnutrition, or experiences diseases such as Crohn's disease and/or celiac disease and/or cancer, or it is caused by drugs such as chemotherapy drugs and/or corticosteroids. Anorexia and/or metabolic bone disease.

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

Preferably, the compositions of the present invention are used to promote healthy bone growth and/or to prevent and reduce the frequency and/or incidence and/or severity and/or duration of bone diseases.

In particular, the compositions of the invention are useful for preventing and reducing the frequency and/or incidence and/or severity and/or duration of fractures.

Preferably, the compositions of the invention are used to promote healthy bone growth. More preferably, the compositions of the invention are used to prevent a decrease in BMD and/or to increase bone mineral content (BMC).

The invention also encompasses the use of a composition of the invention as a synthetic nutrient for promoting healthy bone growth and/or for preventing and/or treating bone diseases, particularly osteomalosis, rickets, osteopenia or osteoporosis disease.

The invention also encompasses the use of the compositions of the invention as synthetic nutrients for preventing BMD reduction and/or increasing BMC.

Such uses encompass compositions, supplements, preferably in unit dosage form.

All of the uses described above are especially intended for use in infants and young children. The compositions and uses of the present invention are particularly suitable for having bone disease risks and having bones A family history of the disease or an infant and young child who have experienced at least one, preferably several, fractures. The compositions and uses of the present invention are particularly suitable for infants and young children who have premature birth or have LBW or experience IUGR, or who suffer from growth retardation due to malnutrition or experience such as Crohn's disease and/or celiac disease and/or cancer A disease, or treatment with a drug such as a chemotherapeutic drug and/or a corticosteroid, results in malabsorption, anorexia, and/or metabolic bone disease. The compositions of the present invention may also be suitable for adolescents or adults who have a risk of bone disease or have experienced at least one, preferably several, fracture episodes, or have premature birth or have LBW or experience IUGR, or have experienced growth retardation due to malnutrition Or experiencing diseases such as Crohn's disease and/or celiac disease and/or cancer, or treatment with drugs such as chemotherapeutic drugs and/or corticosteroids leading to malabsorption, anorexia and/or metabolic bone disease, or in infants Delayed growth during the period and/or during childhood (including adolescence) due to illness or malnutrition or drug use.

The compositions of the present invention are particularly useful in infants and children who have premature birth or have low birth weight or experience intrauterine growth retardation, or have intrauterine malnutrition, or are subject to growth delay.

Without wishing to be bound by theory, the inventors believe that the combination of oligosaccharide mixtures in the above compositions may be beneficial to probiotic growth and/or prevention and/or treatment of bone diseases by probiotic strains and LC-PUFA. The result of a synergistic combination of immune modulator effects triggered by stimulation with a particular oligosaccharide mixture.

Oligosaccharides, LC-PUFAs, and probiotic strains 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 a (premature) infant formula, a supplement, a human milk fortifier, a larger infant formula or a growing milk. Alternatively, for a group of young children, the composition can be, for example, a juice drink or other frozen or shelf stable beverage or soup, or a baby food or baby cereal composition. 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 situation 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 fatty acid ratio.

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 that may be 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 made by source of protein, carbohydrate source (different from oligosaccharide mixture) and fat source (package) Including LC-PUFA) is prepared by blending 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. The water temperature is conveniently 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 small 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 processed through a sterilizer.

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 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.

Supplements can be acceptable to consumers (which are human or animal) (respectively For example, an ingestible carrier or carrier can be added. 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 rhamnosus on rats undergoing IUGR. IUGR is induced by limiting maternal food by 50% from day 11 of pregnancy to delivery.

Method

Sprague-Dawley females and unmatured females mated at the right time were purchased from Charles River, France. The mated female animals are grouped by weight and dispensed according to random or restricted protocols. All animals received a chow diet (Kliba 3437). Restricted females received 50% of the unmarried females from the 11th day of pregnancy until delivery, after which all females were fed ad libitum.

At birth, the juveniles were weighed and assigned to one of three groups (12 males and 12 females) within the first 24 hrs: a) Control: Accepted control tubes for feeding the mothers at will Feeding and control weaning diets; b) IUGR control (IUGRc): Controlled tube feeding for restricted females and juveniles on controlled weaning diet; c) IUGR mixture (IUGRtx): treated tube feeding for restricted females and Treat the pups of the weaning diet.

To minimize maternal effects, the juveniles in the study were randomly selected from each female and assigned to 4 lactating mothers/groups, whereby the siblings of the same litter were randomly assigned to the two IUGR groups. The control pups were breastfed by a mother who was fed ad libitum during pregnancy and the pups in the IUGR group were breastfed by a mother who was restricted during pregnancy.

Each litter was adjusted to 8 pups (4 males and 4 females), of which 6 (3 males and 3 females) from each litter were included in the experimental group. The cub remains with its assigned mother and allows it to suck at random until 21 days of age.

2. Treatment and diet

The following functional ingredients are used in experimental tube feeding and dietary compositions:

a) 98.8% dry matter DDWP composition, the composition of which is detailed in Table 1.

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.

b) NCC 4007 (L. rhamnosus) freeze-dried culture powder with skim milk containing 2.14 x 10 ¹¹ cfu/g.

c) ARASCO® oil containing 43.6% arachidonic acid (ARA) and DHASCO® oil (Martek Biosciences) containing 43.6% docosahexaenoic acid (DHA).

2.1. Tube feeding

7 to 21 days of pups received control or treatment tube feeding. The purpose of treating tube feeding is to provide DDWP of 0.08% by weight (BW), DHA and ARA of 0.03% of BW, and NCC 4007 of 1×10 ⁹ CFU. In placebo tube feeding, DDWP was replaced by malidextrin (Glucidex 12, Roquette), replaced by alpha-linolenic acid (C18:3 n-3) and linoleic acid (C18:2 n-6) DHA and ARA, and the freeze-dried NCC 4007 was replaced by skim milk powder. Other carbohydrates present in the DDWP mixture were also provided in placebo tube feeding (see Table 1).

2.2. Diet

On day 21, the pups were individually housed and weaned to receive controls (control and IUGRc) or experimental diet (IUGRtx) until day 58 (= days after birth). The weaning diet (Tables 2 and 3) was in powder form and stored in vacuum sealed aluminum packaging at 4 °C until use.

¹ 4.53 kcal/g, 18.32% protein ² 4.16 kcal/g, 18.78% protein ³ Smith trading ⁴ Synopharm, Germany, ⁵ Roquette, France. ⁶ Howeg, Switzerland. ⁷ Fluka, ⁸ Merck, Germany. ⁹ Nestlé, Switzerland, ¹⁰ Socochim, Switzerland. ¹¹ Sofinol, Switzerland. ¹² Nutriswiss, Switzerland. ¹³ Martek, USA. ¹⁴ Sabo, Switzerland.

In the experiment, the ratio is as follows: tube feeding: DDWP: ARA: DHA 1:1:1 and diet: DDWP: ARA: DHA 11: 1:1

The nutritional content of the two diets (modified AIN 93 G) is suitable for rat growth requirements. The control diet was supplemented with lactose, glucose, galactose to match the levels of such carbohydrates found in the DDWP product.

The fatty acid profiles of the two diets are balanced to provide similar ratios of n-6/n-3 and similar ratios of saturated, monounsaturated and polyunsaturated fatty acids.

The fatty acid composition of the two diets is nearly identical in terms of fatty acid profile, with the following exceptions: C18: 2 n-6 (linolenic acid) is 16.8% by mass of FA (for controls) and 14.6% by mass (for treatment) , C18: 3 n-3 (α-linolenic acid) is 4.1% by mass of FA (for control) and 2.5% by mass (for treatment), C20: 4 n-9 (ARA) is 0.0 ( For the control) and 2.1% by weight of FA (for treatment) and C22: 6 n-3 (DHA) is 0.0 (for control) and 1.9% by mass of FA (for treatment). The total n-6 of the control and treatment was 18.0% by mass of FA. The total n-3 is 4.1% by mass of FA (for the control) and 4.4% by mass of FA (for treatment), which is quite similar, and the n-6/n-3 ratio is 4.4% by mass of FA (for For comparison) and 4.1% by mass (for treatment), this is similar.

During this time period, animals received gelatin particles with (IUGR-treated) or without (control and IUGR-control) NCC 4007 (1 x ¹⁰⁹ CFU). The granules were replenished each morning and the composition is detailed in Table 4.

Animals were sacrificed on day 58.

3. Skeletal parameters

The nose-valve length was measured before being sacrificed.

After sacrifice, the femur was collected. Carefully remove the bones to complete the periosteum. After measuring the length of the femur, it was stored in gauze soaked in phosphate buffered saline at -80 °C. Each femur was allowed to reach room temperature and then measured.

The bone mineral content (BMC) and bone minerals (BMD) of the entire femur were determined using a dual energy X-ray absorptiometry (DXA). The femur was placed on a reference resected bone plate (purchased by PIXImus) simulating soft tissue and scanned using DXA (PIXImus2; GE-Lunar, Madison, WI). The coefficient of variation for total BMD and BMC is 0.5%.

Figure 1 is a bar graph plotting the results of the experiment on day 58 (= days after birth) of bone mineral content (BMC, g).

Figure 2 is a bar graph plotting experimental results on day 58 (= days after birth) of bone mineral density (BMD, g/cm ² ). Bars indicate the median value of the group ± SE median. Male and female animals are drawn together. The letters "a" and "b" have the following meanings: different letters mean significant differences (p < 0.005).

Figure 3 is a bar graph plotting the results of the experiment on day 58 (= days after birth) of the nose-anal length (or naL, cm).

Figure 4 is a bar graph plotting experimental results for day 58 (= days after birth) of femur length (or fL, mm). Bars indicate the median ± SE median values in these groups. Male and female animals are drawn together. The letters "a" and "b" have the following meanings: different letters mean significant differences (p < 0.005).

According to their results, IUGRc appeared to decrease compared with the control in terms of nasal-valve length (naL), whereas there was no difference between control and IUGRtx (Fig. 3, IUGRc versus control, P <0.01; and IUGRx pair IUGRc, P=0.059). In terms of femur length (fL), IUGRc was reduced compared to control and IUGRtx; there was no difference between control and IUGRtx. (Fig. 4, IUGRc vs. IUGRtx, P=0.027). Similarly, in the case of BMC, IUGRc was reduced compared to controls and IUGRtx (Figure 1, P 0.003); There is no difference between IUGRtx and control. In terms of BMD, IUGRx has increased relative to control and IUGRc (Figure 2, P 0.003).

In the figure: Figure 1 is a bar graph plotting the experimental results for bone mineral concentration (BMC, g).

Figure 2 is a bar graph plotting the experimental results for bone mineral density (BMC, g/cm ² ).

Figure 3 is a diagram showing the results of the experiment on the length of the nose - anus (or naL, cm) Shape chart.

Figure 4 is a bar graph plotting experimental results for femur length (or fL, mm).

Claims (15)

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 healthy bone growth and/or preventing and/or treating bone diseases, preferably osteodystrophy, rickets, osteopenia or osteoporosis. 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, 6Ga1β1,4Glc(=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). 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 , Lactobacillus reuteri , and Bifidobacterium lactis . 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 A formula, a growing milk, a medical food or supplement for clinical nutrition, preferably a premature infant formula, a human milk fortifier or a supplement. The composition of claim 1 or 2 for use in infants and children who have premature birth or have low birth weight or experience intrauterine growth retardation, or have growth stunting due to malnutrition, or experience Diseases such as anorexia, Crohn's disease and/or celiac disease and/or cancer, or treatment with drugs such as chemotherapeutic drugs and/or corticosteroids, resulting in malabsorption, anorexia and/or metabolic bones disease. A composition according to claim 1 or 2 for use in preventing bone mineral density (BMD) reduces and / or is used to increase bone mineral content (BMC). 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 comprising at least one N-acetylated oligosaccharide, at least one sialylation An oligosaccharide and at least one neutral oligosaccharide, the composition for use in the manufacture of a synthetic nutrient for promoting healthy bone growth and/or for preventing and/or treating bone diseases, preferably osteomalosis, rickets, bone mass Reduce or osteoporosis. 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 comprising at least one N-acetylated oligosaccharide, at least one sialylation An oligosaccharide and at least one neutral oligosaccharide for use in the manufacture of a synthetic nutrient for preventing bone mineral density (BMD) reduction and/or for increasing bone mineral content (BMC).