Mature or improved human milk oligosaccharides for supporting sleep patterns

Abstract

The present invention relates to fucosylated human milk oligosaccharides for use in improving sleep maturation in non-infants, such as improving sleep difficulties.

Description

Technical Field

[0001] This invention relates to mature or improved human milk oligosaccharides (HMOs) for use in non-infant subjects to support sleep patterns. In particular, the HMO is a fucoidylated HMO. Background Technology

[0002] Preliminary evidence based on animal studies suggests that human milk oligosaccharides (HMOs) may play an important role in the development of the central nervous system and thus in related functions.

[0003] Reduced sleep is associated with a variety of negative effects, such as decreased cognitive development, mood regulation, and overall health (LJ Meltzer, JA Mindell Psychiatr. Clin. North Am. 29, 1059 (2006)). Specifically, short sleep duration and poor sleep quality are associated with obesity (EMTaveras et al. Arch. Pediatr. Adolesc. Med. 162, 305 (2008)) and behavioral problems (B. Zuckerman et al. Pediatrics 80, 664 (1987)). The most common sleep disorders in infants and children are insomnia-related sleep disorders (difficulty falling asleep at bedtime or being unable to sleep through the night undisturbed). These disorders are estimated to affect 15% to 35% of infants under 24 months of age (France et al. SleepMedicine Reviews 3, 265 (1999)). Furthermore, as early sleep patterns mature, these sleep disorders may appear differently at different ages, shifting from more fragmented non-circadian sleep to long, uninterrupted sleep following a circadian rhythm (TF Anders et al. A longitudinal study of night-time sleep-wake patterns in infants from birth to one year In: JDCall et al. (eds): Frontiers of Infant Psychiatry. New York: Basic Books, pp. 50–166 (1983)).

[0004] Since HMOs are primarily metabolized in the gut, their potential impact on the gut microbiota has been well-established. In contrast, the potential effects of HMOs on brain function development are less studied, with most research in this area focusing on learning and memory. For example, chronic oral administration of 2'-FL to rodents, Vazquez et al. J. Nutr. Biochem. 26, 455 (2015) showed that treated animals exhibited significantly better learning and working memory function. Nevertheless, to our knowledge, this study is the first to report human outcomes and investigate the effects of HMOs on sleep pattern maturation or sleep difficulties.

[0005] Such health benefits need to be delivered in non-infants in a manner that does not cause side effects and / or in a manner that is easy to deliver and generally accepted by patients or healthcare practitioners. They also need to be delivered in a manner that truly keeps the cost of such delivery reasonable and affordable for most people. Summary of the Invention

[0006] In a first aspect, the present invention provides one or more fucoidylated human milk oligosaccharides (HMOs), preferably fucoidylated neutral HMOs, for improving sleep maturation in non-infants, such as improving sleep difficulties.

[0007] In a second aspect, the present invention provides a synthetic composition comprising an effective amount of one or more fucosylated human lactose oligosaccharides (HMOs), preferably fucosylated neutral HMOs, consisting of or substantially consisting of them, for improving sleep maturation in non-infants, such as improving sleep difficulties.

[0008] In one embodiment, the nutritional composition comprises one or more HMOs in a total amount of 50 mg / L to 5000 mg / L, for example 50 mg / L to 2500 mg / L, or 60 mg / L to 2000 mg / L, or 80 mg / L to 1000 mg / L.

[0009] In a third aspect, the present invention provides an encapsulation for improving sleep maturation in non-infants, such as improving sleep difficulties, the encapsulation comprising an effective amount of one or more fucosylated human lactose oligosaccharides (HMOs), preferably fucosylated neutral HMOs, in at least 14 individual daily doses.

[0010] The package preferably contains at least about 21 individual daily doses, such as about 28 daily doses.

[0011] The fourth aspect of the invention is

[0012] - One or more fucoidylated human milk oligosaccharides (HMOs), preferably fucoidylated neutral HMOs.

[0013] - A synthetic composition comprising one or more fucosylated human lactose oligosaccharides (HMOs), preferably fucosylated neutral HMOs, consisting of or substantially consisting of them, or

[0014] - An encapsulation comprising at least 14 individual daily doses of one or more fucoidylated human lactose oligosaccharides (HMOs), preferably fucoidylated neutral HMOs.

[0015] Application in the dietary management of non-infants who have fragmented sleep consisting of frequent short sleep periods and lack of circadian rhythms.

[0016] In a fifth aspect, the present invention provides a method for improving sleep maturity in non-infants, such as improving sleep difficulties, the method comprising administering to the non-infant an effective amount of one or more fucosylated human lactose oligosaccharides (HMOs), preferably fucosylated neutral HMOs.

[0017] Preferably, in any of the foregoing aspects, one or more fucosylated HMOs are fucosylated neutral HMOs.

[0018] Preferably, one or more fucosylated HMOs are administered to the non-infant person for at least one week, more preferably at least two weeks. For example, one or more fucosylated HMOs may be administered to the person for at least four weeks or even longer.

[0019] One or more fucosylated HMOs can be administered to non-infants at a dose of about 0.2 to 10 g per day. In one embodiment, a higher dose may be initially administered to non-infants, followed by a lower dose of one or more fucosylated HMOs. The higher dose is preferably about 0.5 g to about 10 g per day (e.g., about 1 g to about 7.5 g per day), and the lower dose is preferably about 0.1 g to about 5 g per day (e.g., about 0.2 g to about 3 g per day).

[0020] In any of the above aspects, non-infants who have sleep difficulties are, for example, patients with IBS. Detailed Implementation

[0021] As used herein, the following terms have the following meanings.

[0022] "Dietary management" refers to the complete or partial feeding of patients suffering from the following conditions due to illness, ailment, or medical circumstances:

[0023] - The ability to ingest, digest, absorb, metabolize, or excrete ordinary food or certain nutrients or metabolites contained therein is limited, impaired, or disordered.

[0024] - Other medically determined nutritional needs

[0025] (See: Commission Notice on the classification of Food for Special Medical Purposes of the European Commission, Official Journal of the European Union C 401, 25.11.2017, p.10-11).

[0026] "Enterovenous administration" refers to any conventional form of delivery of a composition to a person that results in the deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include via nasogastric or jejunal tube, oral administration, sublingual administration, and rectal feeding.

[0027] "Effective dose" refers to the amount of composition that provides a sufficient amount of HMO in a person to achieve the desired therapeutic outcome. The effective dose may be administered once or multiple times to achieve the desired therapeutic outcome.

[0028] "FODMAP" refers to fermentable oligosaccharides, disaccharides, monosaccharides, and polyols.

[0029] "Human milk oligosaccharides" or "HMOs" refer to a complex carbohydrate found in breast milk (Urashima et al.: Milk Oligosaccharides. Nova Science Publisher (2011); Chen Adv. Carbohydr. Chem. Biochem. 72, 113 (2015)). HMOs have a core structure comprising a reducing lactose unit that can be elongated by one or more β-N-acetyl-lactosamine and / or one or more β-lacto-N-disaccharide units, and this core structure can be partially substituted with α-L-fucopyranosyl and / or α-N-acetyl-neuraminoyl (sialic acid) groups. In this respect, non-acidic (or neutral) HMOs do not contain sialic acid residues, while acidic HMOs have at least one sialic acid residue in their structure. Non-acidic (or neutral) HMOs can be fucosylated or unfucosylated. Examples of neutral, non-fucosylated HMOs include lact-N-tetrasaccharide (LNT), lact-N-neotetrasaccharide (LNnT), lact-N-neohexasaccharide (LNnH), p-lacto-N-neohexasaccharide (pLNnH), p-lacto-N-hexasaccharide (pLNH), and p-N-hexasaccharide (LNH). Examples of neutral, fucosylated HMOs include 2'-fucosylated lactose (2'-FL), lact-N-fucopentose I (LNFP-I), lact-N-difucohexasaccharide I (LNDFH-I), 3-fucosylated lactose (3-FL), difucosylated lactose (DFL), lact-N-fucopentose II (LNFP-II), lact-N-fucopentose III (LNFP-III), and lact-N-difucohexasaccharide III (LNDFH-I). H-III), fucoidan-lacto-N-hexasaccharide II (FLNH-II), lacto-N-fucopentose V (LNFP-V), lacto-N-fucopentose VI (LNFP-VI), lacto-N-difucohexasaccharide II (LNDFH-II), fucoidan-lacto-N-hexasaccharide I (FLNH-I), fucoidan-p-lacto-N-hexasaccharide I (FpLNH-I), fucoidan-p-lacto-N-neohexose II (FpLNnH II), and fucoidan-lacto-N-neohexose (FLNnH).Examples of acidic HMOs include 3'-sialyl lactose (3'-SL), 6'-sialyl lactose (6'-SL), 3-fucosyl-3'-sialyl lactose (FSL), LST a, fucosyl-LST a (FLST a), LST b, fucosyl-LST b (FLST b), LST c, fucosyl-LST c (FLST c), sialyl-LNH (SLNH), sialyl-lacto-N-hexasaccharide (SLNH), sialyl-lacto-N-neohexose I (SLNH-I), sialyl-lacto-N-neohexose II (SLNH-II), and disialyl-lacto-N-tetrasaccharide (DSLNT).

[0030] Irritable bowel syndrome (IBS) is a functional bowel disorder characterized by recurrent abdominal pain associated with bowel movements or changes in bowel habits. Disorders of bowel habits (constipation, diarrhea, or a combination of both) and symptoms of abdominal bloating / distension are typical. For diagnosis according to the Rome IV criteria, the patient must have experienced recurrent abdominal pain on average for at least one day per week over the past 3 months, and the pain must be associated with two or more of the following criteria: (1) associated with bowel movements; (2) associated with changes in bowel frequency; and (3) associated with changes in stool form (appearance). Symptoms should have occurred at least 6 months prior to diagnosis and have been present for the most recent 3 months.

[0031] "Microbiome," "microbial community," and "microbial group" refer to the living microbial community that usually resides in the organs or parts of the body, especially the human gastrointestinal tract. The most important members of the gastrointestinal microbiota include the phyla Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, Synergistetes, Verrucomicrobia, Fusobacteria, and Euryarchaeota. At the genus level, genera such as Bacteroides, Faecalibacterium, Bifidobacterium, Roseburia, Alistipes, Collinsella, Blautia, Coprococcus, Ruminococcus, Eubacterium, and Dorea exist. At the species level, Bacteroides... The gut microbiota includes mucosa-associated microbes located on or attached to the mucosal layer covering the gastrointestinal epithelium, as well as lumen-associated microbes found in the gastrointestinal lumen. These microbes include *Alistipes putredinis*, *Parabacteroides merdae*, *Ruminococcus bromii*, *Dorea longicatena*, *Bacteroides caccae*, *Bacteroides thetaiotaomicron*, *Eubacterium hallii*, *Ruminococcus torques*, *Faecalibacterium prausnitzii*, *Ruminococcus lactaris*, *Collinsella aerofaciens*, *Doreaformicigenerans*, *Bacteroides vulgatus*, and *Roseburia intestinalis*.

[0032] "Regulating the microbiome" refers to exerting altering or controlling effects on the microbiome, such as effects that increase the native gut abundance of Bifidobacteria, Barnesiella, Faecalibacterium, and / or butyrate-producing bacteria. In another instance, such an effect may lead to a decrease in the gut abundance of Ruminococcus gnavus and / or Proteobacteria. Proteobacteria is a phylum of Gram-negative bacteria that includes a wide variety of pathogenic bacteria, such as Escherichia coli, Salmonella, Vibrio, Helicobacter pylori, Yersinia, and many other well-known genera.

[0033] "Non-infant person" or "non-infant" refers to a person aged 3 years or older. Non-infant persons can be children, adolescents, adults, or the elderly, with adolescents, adults, or the elderly being preferred, and adults or the elderly being even more preferred.

[0034] "Oral administration" refers to any conventional form of delivery of the composition to a person through the mouth. Therefore, oral administration is a form of enteral administration.

[0035] "Sleep maturity or sleep pattern maturity" refers to improving sleep difficulties, such as the development of nighttime and daytime sleep patterns from more fragmented sleep consisting of frequent short sleeps and a lack of circadian rhythm to a pattern of long, uninterrupted nighttime sleep and limited daytime sleep with a circadian rhythm.

[0036] "Synthetic composition" refers to an artificially prepared composition, preferably a composition containing at least one compound produced in vitro through chemical reaction, enzymatic reaction, or recombination. Synthetic compositions typically contain one or more HMOs. Furthermore, in some embodiments, the synthetic composition may contain one or more nutritional or pharmaceutically active ingredients that do not adversely affect the efficacy of the HMO. Some non-limiting embodiments of the synthetic compositions of the present invention are described below.

[0037] The fucosylated HMOs used in the synthetic compositions, encapsulations, and methods of the present invention can be isolated or enriched from the milk secreted by mammals, including but not limited to humans, cattle, sheep, pigs, or goats, by well-known methods. HMOs can also be produced by well-known methods using microbial fermentation, enzymatic processes, chemical synthesis, or combinations of these techniques. As an example, 2'-FL can be prepared chemically as described in WO 2010 / 115934 and WO 2010 / 115935. As an example of enzymatic production, sialylated oligosaccharides can be prepared as described in WO 2012 / 007588, fucosylated oligosaccharides can be prepared as described in WO 2012 / 127410, and advantageously, various mixtures of human milk oligosaccharides can be prepared as described in WO 2012 / 156897 and WO 2012 / 156898. Biotechnical methods describing the preparation of human lactose oligosaccharides with a core optionally substituted with fucose or sialic acid can be found in WO 01 / 04341 and WO 2007 / 101862. Specifically, WO2015 / 032413, WO2015 / 032412, and WO2016 / 095924 describe biotechnological methods for the production of 2'-FL and DFL.

[0038] Fucosylated neutral HMOs can be supplemented with other human milk oligosaccharides. For example, fucosylated neutral HMOs can be supplemented with non-fucosylated HMOs, such as LNT and / or LNnT.

[0039] Fucosylated neutral HMOs, optionally combined with non-fucosylated HMOs, can be used as is (pure) without any carriers and / or diluents. In another embodiment, fucosylated neutral HMOs, optionally combined with non-fucosylated HMOs, are used in synthetic compositions with one or more inert carriers / diluents acceptable in nutritional or pharmaceutical compositions, such as solvents (e.g., water, water / ethanol, oil, water / oil), dispersants, coatings, absorption enhancers, controlled-release agents, and inert excipients (e.g., starch, polyols, granulators, microcrystalline cellulose, diluents, lubricants, binders, disintegrants). These compositions are free of prebiotics and / or probiotics [i.e., synthetic compositions consisting of a) fucosylated neutral HMOs, optionally combined with non-fucosylated HMOs, and b) one or more inert carriers / diluents acceptable in nutritional or pharmaceutical compositions]. In another embodiment, fucosylated neutral HMOs, optionally combined with non-fucosylated HMOs, are used in synthetic pharmaceutical or nutritional compositions that may contain prebiotics and / or probiotics.

[0040] In this regard, the following implementation methods are used to improve sleep maturation in non-infants, such as improving sleep difficulties, and particularly include:

[0041] -A single HMO selected from 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, and LNFP-VI.

[0042] -Two HMOs selected from 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, and LNFP-VI.

[0043] -A fucosylated neutral HMO selected from 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, and LNFP-VI, and at least one other non-fucosylated neutral HMO.

[0044] - A synthetic composition comprising, or substantially comprising, a fucosylated neutral HMO selected from 2'-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, LNFP-VI, and combinations thereof.

[0045] In a preferred embodiment, the HMO is 2'-FL. In other preferred embodiments, the HMO is DFL. In other preferred embodiments, the HMO is 3-FL. In other preferred embodiments, the HMO is LNFP-I. In other preferred embodiments, the HMO is a combination of 2'-FL and DFL. In other preferred embodiments, the HMO is a combination of 2'-FL and LNFP-I. In other preferred embodiments, the HMO is a combination of 2'-FL and (LNT and / or LNnT) [i.e., 2'-FL and LNT; 2'-FL and LNnT; 2'-FL, LNT, and LNnT]. In other preferred embodiments, the HMO is a combination of 2'-FL, DFL, and (LNT and / or LNnT) [i.e., 2'-FL, DFL, and LNT; 2'-FL, DFL, and LNnT; 2'-FL, DFL, LNT, and LNnT]. More preferably, the HMO is 2'-FL, optionally combined with LNnT.

[0046] Regarding the preferred and more preferred embodiments described above, the preferred non-infant population is selected from children, adolescents, adults, and the elderly; more preferably from adolescents, adults, and the elderly; and even more preferably from adults and the elderly. In one embodiment of the non-infant population, the child, adolescent, adult, or elderly, more preferably from adolescents, adults, or the elderly, and even more preferably from adults or the elderly, is a patient with irritable bowel syndrome (IBS).

[0047] Furthermore, the preferred embodiments of the daily dose included in the package according to the third aspect are those disclosed above.

[0048] Furthermore, the preferred embodiments of the use of the fourth aspect in the dietary management of non-infant individuals are those disclosed above.

[0049] Furthermore, the preferred embodiments of the method according to the fifth aspect are those disclosed above.

[0050] The synthetic composition can be in the form of a nutritional composition. For example, a nutritional composition can be a food composition, rehydration solution, medical food, or food for a specific medical purpose, nutritional supplement, etc. The nutritional composition may contain sources of protein, lipids, and / or digestible carbohydrates, and may be in powder or liquid form. The composition may be designed as a sole source of nutrition or as a nutritional supplement.

[0051] Suitable protein sources include milk protein, soy protein, rice protein, pea protein, and oat protein, or mixtures thereof. Milk protein can be milk protein concentrate, milk protein isolate, whey protein, or casein, or a mixture of both. Protein can be partially hydrolyzed or extensively hydrolyzed whole or hydrolyzed protein. Hydrolyzed protein has the advantage of being easier to digest, which is important for people with gastrointestinal inflammation or damage. Protein can also be provided in the form of free amino acids. Protein can constitute about 5% to about 30% of the energy in a nutritional composition, typically about 10% to 20%.

[0052] Protein sources can be sources of glutamine, threonine, cysteine, serine, proline, or combinations of these amino acids. Glutamine sources can be glutamine dipeptides and / or glutamine-rich proteins. Glutamine can be included because intestinal epithelial cells use glutamine as an energy source. Threonine, serine, and proline are important amino acids for the production of mucin. Mucin lines the gastrointestinal tract and can improve intestinal barrier function and mucosal healing. Cysteine ​​is a major precursor to glutathione, which is key to the body's antioxidant defense.

[0053] Suitable digestible carbohydrates include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup, corn syrup solids, high-fructose corn syrup, rice-derived carbohydrates, pea-derived carbohydrates and potato-derived carbohydrates, tapioca starch, sucrose, glucose, fructose, lactose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), or mixtures thereof. In one embodiment, the composition has reduced or no added lactose or other FODMAP carbohydrates. Generally, digestible carbohydrates provide about 35% to about 55% of the energy of the nutritional composition. A particularly suitable digestible carbohydrate is low-dextrose equivalent (DE) maltodextrin.

[0054] Suitable lipids include medium-chain triglycerides (MCTs) and long-chain triglycerides (LCTs). Preferably, the lipids are a mixture of MCTs and LCTs. For example, MCTs may contain about 30% to about 70% lipids by weight, more specifically, about 50% to about 60% by weight. MCTs have the advantage of being easier to digest, which is important for people with gastrointestinal inflammation or damage. Typically, lipids provide about 35% to about 50% of the energy of a nutritional composition. Lipids may contain essential fatty acids (ω-3 and ω-6 fatty acids). Preferably, these polyunsaturated fatty acids provide less than about 30% of the total energy from the lipid source.

[0055] Suitable sources of long-chain triglycerides include rapeseed oil, sunflower oil, palm oil, soybean oil, milk fat, corn oil, high-oleic acid, and soybean lecithin. Fractionated coconut oil is a suitable source of medium-chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a ratio of polyunsaturated fatty acids ω-6 (n-6) to ω-3 (n-3) of about 4:1 to about 10:1. For example, the ratio of n-6 to n-3 fatty acids can be about 6:1 to about 9:1.

[0056] Nutritional compositions may also contain vitamins and minerals. If the nutritional composition is intended to be the sole source of nutrition, it preferably contains a complete spectrum of vitamins and minerals. Examples of vitamins include vitamins A, B complex (such as B1, B2, B6, and B12), C, D, E, and K, niacin, and acidic vitamins such as pantothenic acid, folic acid, and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium, and boron.

[0057] The nutritional composition may also contain carotenoids, such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoids contained may vary between about 0.001 μg / ml and about 10 μg / ml. The lutein content may be about 0.001 μg / ml to about 10 μg / ml, preferably about 0.044 μg / ml to about 5 μg / ml. The lycopene content may be about 0.001 μg / ml to about 10 μg / ml, preferably about 0.0185 μg / ml to 5 μg / ml. Beta-carotene may be contained between about 0.001 μg / ml and about 10 mg / ml, for example, about 0.034 μg / ml to about 5 μg / ml.

[0058] The nutritional composition preferably also contains sodium at a reduced concentration, for example, from about 300 mg / L to about 400 mg / L. The remaining electrolytes can be present at the required concentrations without placing an excessive solute burden on renal function. For example, potassium is preferably present in the range of about 1180 to about 1300 mg / L, and chloride is preferably present in the range of about 680 to about 800 mg / L.

[0059] Nutritional compositions may also contain various other conventional ingredients, such as preservatives, emulsifiers, thickeners, buffers, fiber, and prebiotics (e.g., fructooligosaccharides, galactooligosaccharides), and probiotics (e.g., *Bifidobacterium animalis* subsp. *lactis* BB-12, *Bifidobacterium lactis* HN019, *Bifidobacterium lactis* Bi07, *Bifidobacterium infantis* ATCC). 15697, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus HNOOl, Lactobacillus acidophilus LA-5, Lactobacillus acidophilus NCFM, Lactobacillus fermentum CECT5716, Bifidobacterium longum BB536, Bifidobacterium longum AH1205, Bifidobacterium longum AH1206, Bifidobacterium breve M-16V, Lactobacillus reuteri ATCC 55730, Lactobacillus reuteri ATCC PTA-6485, Lactobacillus reuteri DSM17938), antioxidant / anti-inflammatory compounds include tocopherols, carotenoids, ascorbic acid / vitamin C, ascorbate palmitate, polyphenols, glutathione and superoxide dismutase (melon), other bioactive factors (such as growth hormones, cytokines, TFG-β), colorants, fragrances and stabilizers, lubricants, etc.

[0060] In one embodiment, the nutritional composition is gluten-free.

[0061] The nutritional composition can be formulated as a soluble powder, liquid concentrate, or ready-to-use preparation. It can be administered to those in need via nasogastric tube or orally. Various flavorings, fibers, and other additives may also be present.

[0062] Nutritional compositions can be prepared using any commonly used manufacturing techniques for preparing solid or liquid nutritional compositions. For example, the composition can be prepared by combining various feedstock liquids. A fat-encapsulated protein feedstock liquid can be prepared by heating and mixing a lipid source, followed by the addition of an emulsifier (e.g., lecithin), fat-soluble vitamins, and at least a portion of a protein source while heating and stirring. A carbohydrate feedstock liquid can then be prepared by adding minerals, trace and ultra-trace minerals, thickeners, or suspending agents to water while heating and stirring. The resulting solution is kept under continuous heating and stirring for 10 minutes before adding carbohydrates (e.g., HMOs and digestible carbohydrate sources). The resulting feedstock liquids are then mixed together while heating and stirring, and the pH is adjusted to 6.6–7.0. The composition is then subjected to a short-time high-temperature treatment during which it is heat-treated, emulsified, and homogenized, followed by cooling. Water-soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavorings are added, and water is added to achieve the desired total solids content.

[0063] For liquid products, the resulting solution can then be aseptically packaged to form an aseptically packaged nutritional composition. In this form, the nutritional composition can be in a ready-to-eat or concentrated liquid form. Alternatively, the composition can be spray-dried, processed, and packaged into a reconstitutable powder.

[0064] When the nutritional product is a ready-to-eat nutritional liquid, the total concentration of sialylated and / or non-fucosylated neutral HMOs in the liquid is preferably from about 0.1% to about 1.5% by weight, including about 0.2% to about 1.0%, for example, about 0.3% to about 0.7%. When the nutritional product is a concentrated nutritional liquid, the total concentration of sialylated and / or non-fucosylated neutral HMOs in the liquid is preferably from about 0.2% to about 3.0% by weight, including about 0.4% to about 2.0%, for example, about 0.6% to about 1.5%.

[0065] In another embodiment, the nutritional composition is in unit dosage form. Unit dosage form may contain an acceptable food-grade carrier, such as a phosphate-buffered saline solution, an ethanol-water mixture, water, and an emulsion (such as an oil / water or water / oil emulsion), as well as various wetting agents or excipients. Unit dosage form may also contain other materials that will not cause adverse, allergic, or other unwanted reactions when administered to humans. The carrier and other materials may include solvents, dispersants, coatings, absorption enhancers, controlled-release agents, and one or more inert excipients, such as starch, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrants. Preferably, the carrier and other materials have low or no FODMAP content. Preferably, the unit dosage form primarily comprises sialylated and / or non-fucosylated neutral HMOs and a minimal amount of binders and / or excipients. Unit dosage forms are particularly suitable when nutrition is incomplete or not intended as the sole source of nutrition.

[0066] The unit dosage forms of the present invention can be administered orally, for example, in the form of tablets, capsules, or pills containing a predetermined amount of the mixture, or in the form of powders or granules containing a predetermined concentration of the mixture, or in the form of gels, pastes, solutions, suspensions, emulsions, syrups, pills, pharmaceutical sugars, or slurries containing a predetermined concentration of the mixture as an aqueous or non-aqueous liquid. Orally administered compositions may include one or more binders, lubricants, inert diluents, flavoring agents, and humectants. Orally administered compositions, such as tablets, may optionally be coated and may be formulated to provide sustained or controlled release of sialylated and / or non-fucosylated neutral HMOs.

[0067] The unit dosage form of the present invention can also be administered via nasogastric tube or directly into the gastrointestinal tract or stomach.

[0068] The unit dosage form of the present invention may also include therapeutic agents, such as antibiotics, probiotics, analgesics and anti-inflammatory agents.

[0069] The appropriate dosage of this composition for humans can be determined in a conventional manner based on factors such as the individual's condition, immune status, weight, and age. In some cases, the concentration of the dosage is similar to that of the fucosylated neutral HMO in human breast milk. The required amount is typically in the range of about 0.1 g to about 10 g per day, and in some embodiments, about 0.2 g to about 7.5 g per day, for example, about 1 g to about 5 g per day. An appropriate dosage regimen can be determined by methods known to those skilled in the art.

[0070] In another embodiment, fucosylated neutral HMOs can be formulated into pharmaceutical compositions. The pharmaceutical compositions may comprise pharmaceutically acceptable carriers, such as phosphate-buffered saline solutions, ethanol-water mixtures, water, and emulsions, such as oil / water or water / oil emulsions, as well as various wetting agents or excipients. The pharmaceutical compositions may also comprise other materials that will not produce adverse, allergic, or other unwanted reactions when administered to humans. Carriers and other materials may include solvents, dispersants, coatings, absorption enhancers, controlled-release agents, and one or more inert excipients, such as starch, granulating agents, microcrystalline cellulose, diluents, lubricants, binders, and disintegrants. Preferably, the carriers and other materials have low or no FODMAP content.

[0071] Pharmaceutical compositions can be administered orally, for example, in the form of tablets, capsules, or pills containing a predetermined amount of the mixture, or in the form of powders or granules containing a predetermined concentration of the mixture, or in the form of gels, pastes, solutions, suspensions, emulsions, syrups, pills, saccharides, or liquids as aqueous or non-aqueous liquids containing a predetermined concentration of the mixture. Orally administered compositions may include one or more binders, lubricants, inert diluents, flavoring agents, and humectants. Orally administered compositions, such as tablets, may optionally be coated and may be formulated to provide sustained, delayed, or controlled release of the mixture described herein.

[0072] The drug composition can also be administered via rectal suppositories, aerosol tubes, nasogastric tubes, or direct infusion into the gastrointestinal tract or stomach.

[0073] The pharmaceutical composition may also include therapeutic agents, such as antibiotics, probiotics, analgesics, and anti-inflammatory agents.

[0074] The appropriate dosage of these compositions for humans can be determined in a conventional manner based on factors such as disease condition, immune status, weight, and age. In some cases, the concentration of the dosage is similar to that of fucosylated neutral HMOs in human breast milk. The required amount is typically in the range of about 0.1 g to about 10 g per day, and in some embodiments, about 0.2 g to about 7.5 g per day, for example, about 1 g to about 5 g per day. A suitable dosage regimen can be determined by conventional methods.

[0075] The amount of fucoidylated neutral HMO to be administered will vary depending on factors such as the risk and severity of the underlying condition, any other medical conditions or diseases, age, the form of the composition, and other medications administered. Furthermore, the amount may vary depending on whether the combination is used to provide a direct effect (when the dose may be higher) or whether the combination is used for secondary prevention / maintenance (when the dose may be lower). However, the required amount can be readily determined by a practicing physician and is generally in the range of about 0.1 g to about 10 g per day, and in some embodiments, about 0.2 g to about 7.5 g per day, for example, about 1 g to about 5 g per day. A suitable dose can be determined based on several factors, including, for example, weight and / or condition, the severity of the underlying condition being treated or prevented, other ailments and / or diseases, the incidence and / or severity of side effects, and the method of administration. A suitable dose range can be determined by methods known to those skilled in the art. In the initial treatment phase, the dose may be higher (e.g., 0.5 g to 10 g per day, preferably 1 g to 7.5 g per day). During the maintenance phase, the dosage can be reduced (e.g., 0.1g to 5g per day, preferably 0.2g to 3g per day).

[0076] Example

[0077] The present invention will now be further illustrated by the following embodiments:

[0078] Example 1 – Nutritional Composition

[0079] The ready-to-eat nutritional composition is prepared from water, maltodextrin, enzymatically hydrolyzed whey protein (from milk), medium-chain triglycerides (from coconut and / or palm kernel oil), corn starch, soybean oil, soybean lecithin, 2'-FL, magnesium chloride, calcium phosphate, guar gum, sodium ascorbate, potassium citrate, sodium phosphate, calcium citrate, choline chloride, potassium chloride, sodium citrate, magnesium oxide, taurine, L-carnitine, α-tocopherol acetate, zinc sulfate, ferrous sulfate, nicotinamide, calcium pantothenate, vitamin A palmitate, citric acid, manganese sulfate, pyridoxine hydrochloride, vitamin D3, copper sulfate, thiamine mononitrate, riboflavin, β-carotene, folic acid, biotin, potassium iodide, chromium chloride, sodium selenate, sodium molybdate, phytonabinone, and vitamin B12.

[0080] The composition has a caloric density of 1.0 kcal / ml, with the following caloric distribution (% of kcal): protein: 16%, carbohydrates: 51%, fat: 33%. The NPC:N ratio of the protein source is 131:1. The MCT:LCT ratio is 70:30, and the n6:n3 ratio is 7.4:1. Without added flavoring, the osmotic pressure (mOsm / kg water) is 270. This composition contains 85% water and meets 100% RDI for 22 key micronutrients per 1500 mL.

[0081] This composition has a balanced peptide profile that promotes gastrointestinal absorption and integrity, and the enzymatically hydrolyzed 100% whey protein promotes gastric emptying. MCT reduces the likelihood of fat malabsorption. This composition is nutritionally complete and can be used for tube feeding or oral supplementation.

[0082] Example 2 – Tablet Composition

[0083] The tablets are prepared from 2'-FL (or 2'-FL:LNnT 4:1 by weight), hydroxypropyl methylcellulose, sodium alginate, gum, microcrystalline cellulose, colloidal silica, and magnesium stearate. All raw materials except magnesium stearate are placed in a high-shear granulator and premixed. Water is sprayed onto the premix while mixing continues at 300 rpm. The granules are transferred to a fluidized bed dryer and dried at 75°C. The dried powder is sieved and graded using a grinder. The resulting powder is then lubricated with magnesium stearate and compressed into tablets. Each tablet contains 325 mg of 2'-FL (or 2'-FL:LNnT 4:1 by weight). The weight of each tablet is 750 mg.

[0084] Example 3 – Capsule Composition

[0085] Capsules are prepared by filling approximately 1 g of 2'-FL (or 2'-FL:LNnT 4:1 by weight) into 000-gelatin capsules using a filling machine. The capsules are then closed. The 2'-FL (or 2'-FL:LNnT 4:1 by weight) is in free-flowing powder form.

[0086] Example 4 – Nutritional Composition

[0087] Introduce 2'-FL (or 2'-FL:LNnT 4:1 by weight) into a rotary mixer. Introduce 0.25 w% silica into the mixer and mix for 10 minutes. Then, coagulate the mixture in a fluidized bed and fill it into 5 g rod-shaped encapsulations, and seal the encapsulations.

[0088] Example 5 – Human Trial

[0089] In a single-arm, open-label trial, 317 male and female patients who completed the baseline procedure were recruited to participate in the study. Patients were eligible to participate if they were 18 years of age or older; met the diagnostic criteria for Rome type IV irritable bowel syndrome (IBS); had English reading, speaking, and comprehension abilities; and were able and willing to understand and comply with the study procedures. Patients were excluded if they had participated in a clinical study one month prior to their screening visit; had a physician diagnosis of celiac disease, Crohn's disease, ulcerative colitis, diverticulitis, inflammatory bowel disease, or Clostridium difficile infection; were pregnant, breastfeeding, or wishing to become pregnant during the study period; or were deemed unsuitable for the study for any reason in the judgment of the field investigator or principal investigator.

[0090] During the initial screening visit, patients were given written and verbal information about the study and were asked to sign an informed consent form. Patients were assessed through a comprehensive review of their clinical history, and the diagnostic criteria for IBS were evaluated. Upon enrollment in the trial, patients completed an electronic baseline survey.

[0091] The study intervention lasted 12 weeks. After the baseline survey, the survey product for the first eight weeks was sent to patients via email, while the remaining four weeks' worth of the product was sent after the follow-up survey four weeks after the intervention. The survey product consisted of a 4:1 weight ratio of 2'-FL and LNnT, administered daily at a dose of 5 grams, and was provided as a white powder in a single-use stick package. During the study, patients were instructed not to alter their normal diet. During the intervention, recruited patients completed three online follow-up surveys every four weeks. The online survey consisted of several validated questionnaires, including the PHQ-12 (a 12-item Patient Health Questionnaire; a tool for quantifying the severity of non-gastrointestinal physical symptoms).

[0092] The question, "How much have you been bothered by sleep difficulties in the past 4 weeks?" was rated from 0 = 'Not bothered at all' to 2 = 'Very bothered'. Patients reported significant improvement in 'sleep difficulties' symptoms after 4 weeks of intervention, and a 35% improvement after 12 weeks (see table below).

[0093] average 1.04 0.60 0.65 0.67 Standard deviation 0.72 0.68 0.71 0.66

[0094] It can be concluded that daily supplementation with a combination of 5 grams of 2'-FL and LNnT significantly improves patients' sleep problems.

Claims

1. Use of one or more fucoidylated human milk oligosaccharides (HMOs) in the preparation of medicaments for improving sleep in non-infant mature individuals. The aforementioned sleep maturation includes the development of nighttime and daytime sleep patterns from more fragmented sleep consisting of frequent short sleep periods and a lack of circadian rhythms to a pattern of long, uninterrupted nighttime sleep and limited daytime sleep with a circadian rhythm. The HMO mentioned therein is 2'-FL; The HMO is a combination of 2'-FL and (LNT and / or LNnT); or The HMO is a combination of 2'-FL, DFL, and (LNT and / or LNnT).

2. The use according to claim 1, wherein the drug is used to improve sleep difficulties in non-infants.

3. The use according to claim 1 or 2, wherein the HMO is administered to the non-infant person for a period of at least one week.

4. The use according to claim 1 or 2, wherein the HMO is administered to the non-infant person for a period of at least 2 weeks.

5. The use according to claim 1 or 2, wherein the non-infant is given a daily dose of 0.1 g to 10 g of HMO.

6. The use according to claim 5, wherein the non-infant is administered HMO at a daily dose of 0.2 g to 7.5 g.

7. The use according to claim 5, wherein the daily dose of HMO is administered to the non-infant person at a dose of 1 g to 5 g.

8. The use according to claim 1 or 2, wherein a higher dose is initially administered to the non-infant person, followed by a lower dose.

9. The use according to claim 8, wherein the higher dose is 0.5 g to 10 g per day and the lower dose is 0.1 g to 5 g per day.

10. The use according to claim 9, wherein the higher dose is 1 g to 7.5 g per day.

11. The use according to claim 9, wherein the lower dose is 0.2 g to 3 g per day. 12.-One or more fucoidylated human milk oligosaccharides (HMOs), - A synthetic composition comprising one or more fucoidylated human lactose oligosaccharides (HMOs), or - An encapsulation comprising at least 14 individual daily doses of one or more fucoidylated human lactose oligosaccharides (HMOs). Non-therapeutic use in the dietary management of non-infant individuals who have fragmented sleep consisting of frequent short sleep periods and lack of circadian rhythms. The HMO mentioned therein is 2'-FL; The HMO is a combination of 2'-FL and (LNT and / or LNnT); or The HMO is a combination of 2'-FL, DFL, and (LNT and / or LNnT).

13. The use according to claim 1 or 2, or the use according to claim 12, wherein the non-infant is a child, adolescent, adult, or elderly person.

14. The use according to claim 1 or 2, or the use according to claim 12, wherein the non-infant is an IBS patient.