阅读说明：本技术 治疗小麦敏感性的hmo混合物 (HMO mixtures for treating wheat sensitivity ) 是由 L·K·比格斯奈斯 B·麦康奈尔 于 2018-11-30 设计创作，主要内容包括：本发明涉及人乳寡糖(HMO)、包含HMO的合成组合物用于人的非乳糜泻小麦和/或麸质敏感性的治疗、二级预防和/或诱导耐受性以及其方法。(The present invention relates to Human Milk Oligosaccharides (HMOs), synthetic compositions comprising HMOs for use in the treatment, secondary prevention and/or induction of tolerance in non-celiac wheat and/or gluten sensitivity in humans and methods thereof.)

1. A Human Milk Oligosaccharide (HMO) for use in:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans.

2. A synthetic composition for:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans,

the composition comprises at least one Human Milk Oligosaccharide (HMO).

3. Synthetic composition for use according to claim 2, containing at least one HMO in an amount of from 1g to 15g, preferably from 2g to 10g, more preferably from 3g to 7 g.

4. A package comprising an effective amount of at least one Human Milk Oligosaccharide (HMO) in at least 14 independent daily doses for:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans.

5. A package for use according to claim 4, wherein each daily dose contains from about 1g to about 15g, preferably from about 2g to about 10g, of at least one HMO.

6. -one or more Human Milk Oligosaccharides (HMOs),

-a synthetic composition comprising one or more Human Milk Oligosaccharides (HMOs), or

-a package comprising an effective amount of at least one or more Human Milk Oligosaccharides (HMOs) of at least 14 independent daily doses

Use in the dietary management of patients suffering from non-celiac wheat and/or gluten sensitivity.

7. Human milk oligosaccharide for use according to claim 1, synthetic composition for use according to claim 2 or 3, encapsulate for use according to claim 4 or 5, or use according to claim 6, wherein the human milk oligosaccharide comprises 2 ' -FL, 3-FL, DFL, LNT, LNnT, 3 ' -SL, 6 ' -SL, LNFP-I or a mixture thereof.

8. Human milk oligosaccharide for use according to claim 1, synthetic composition for use according to claim 2 or 3, encapsulate for use according to claim 4 or 5, or use according to claim 6, wherein the human milk oligosaccharide comprises, consists of or consists essentially of at least one neutral HMO.

9. The human milk oligosaccharide for use according to claim 8, the synthetic composition for use, the encapsulate for use or the use, wherein the neutral HMO is a mixture of fucosylated HMO and non-fucosylated HMO.

10. The human milk oligosaccharide for use according to claim 9, the synthetic composition for use, the encapsulate for use or the use, wherein the cocktail comprises, consists of, or consists essentially of at least one of 2' -FL and DFL and at least one of LNnT and LNT.

11. The human milk oligosaccharide for use according to claim 10, the synthetic composition for use, the encapsulate for use or the use, wherein the mixture comprises, consists or consists essentially of 2' -FL and LNnT, preferably in a mass ratio of about 4:1 to 1: 1.

12. A method for the treatment of non-celiac wheat and/or gluten sensitivity in a human, the method comprising administering to the human an effective amount of at least one Human Milk Oligosaccharide (HMO).

13. The method according to claim 12, wherein the HMO is administered to the human in an amount of 3g to 15g per day, such as about 4g to about 7.5g per day.

14. The method of claim 12 or 13, wherein the HMO is administered to the human for at least 7 consecutive days.

15. A method for secondary prevention of non-celiac wheat and/or gluten sensitivity in a human, the method comprising administering to the human an effective amount of at least one Human Milk Oligosaccharide (HMO).

16. A method for inducing wheat and/or gluten tolerance in a patient suffering from non-celiac wheat and/or gluten sensitivity, the method comprising administering to the patient an effective amount of at least one Human Milk Oligosaccharide (HMO).

17. The method of claim 16, wherein HMO is administered to the patient upon consumption of a gluten-containing grain.

18. The method according to any one of claims 15 to 17, wherein 2g to 7.5g HMO are administered to the human per day, such as about 2g to about 5g per day.

19. The method of any one of claims 15 to 18, wherein the HMO is administered to the human for at least 4 consecutive weeks.

20. The method according to any one of claims 12-19, wherein the human milk oligosaccharide comprises 2 ' -FL, 3-FL, DFL, LNT, LNnT, 3 ' -SL, 6 ' -SL, LNFP-I, or a mixture thereof.

21. The method according to any one of claims 12 to 19, wherein the human milk oligosaccharide comprises, consists of or consists essentially of at least one neutral HMO.

22. The method of claim 21, wherein the neutral HMOs are a mixture of fucosylated HMOs and nonfucosylated HMOs.

23. The method of claim 22, wherein the mixture comprises, consists of, or consists essentially of at least one of 2' -FL and DFL and at least one of LNnT and LNT.

24. The method of claim 23, wherein the mixture comprises, consists of, or consists essentially of 2' -FL and LNnT, preferably in a mass ratio of about 4:1 to 1: 1.

25. The method according to any one of the preceding claims, comprising a first stage for treating the non-celiac wheat and/or gluten sensitivity and a second stage for inducing wheat and/or gluten tolerance or secondary prevention.

26. The method of claim 25, wherein a higher dose of HMO is administered to the human during the first phase and then a lower dose of HMO is administered to the human during the second phase.

27. The method of claim 26, wherein the higher dose is about 3g to about 15g per day, such as about 4g to about 7.5g per day, and the lower dose is about 2g to about 7.5g per day, such as about 2g to about 5g per day.

28. The method of any one of claims 25 to 27, wherein the duration of the first phase is at least 7 consecutive days and the duration of the second phase is at least four consecutive weeks.

Technical Field

The present invention relates to methods, compounds and compositions for preventing and/or treating non-celiac wheat susceptibility.

Background

Non-celiac wheat sensitivity, also known as non-celiac gluten sensitivity, is a gluten-related disorder that accompanies celiac disease and wheat allergy (Leccioli et al nutriments 9,1203 (2017)). Non-celiac wheat sensitivity is a non-allergic and non-autoimmune condition in which consumption of wheat and/or gluten results in similar symptoms as other gluten-related conditions. This condition is considered to be wheat and/or gluten sensitivity, as symptoms can be alleviated by wheat and/or gluten removal and reappear after wheat and/or gluten reintroduction. However, patients with this condition do not exhibit the characteristic autoimmune or allergic markers associated with celiac disease or gluten allergy. Nevertheless, the clinical symptoms are similar to those of celiac disease and gluten allergy. The most common gastrointestinal symptoms include abdominal distension, abdominal pain, epigastric pain, diarrhea, and constipation. Parenteral symptoms include tiredness, headache, anxiety, "blurry head" or difficulty concentrating, depression and rash. These symptoms can occur within hours to days after exposure to wheat and/or gluten, and then can disappear after the wheat and/or gluten is removed.

The etiology of non-celiac wheat susceptibility is unclear. As seen in celiac disease and gluten allergy, this condition may involve a number of causes. The initial predisposition involves primarily exposure of the intestinal epithelium to wheat and/or gluten, resulting in immune-mediated and/or non-immune-mediated responses. Due to the lack of evidence of T cell involvement and significant Toll-like receptor (TLR) involvement, this condition may be more of an innate immune response than an adaptive immune response.

Since no specific cause has been identified, this condition may involve a different mechanism than celiac disease and gluten allergy. Gluten, the cause of celiac disease, may be an important cause of the condition, but it is increasingly suspected as a major or root cause. Stimulation of some other food sources may also be an important cause. These include alpha amylase/trypsin inhibitors (ATI), fermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPS) and other short chain fructans. In particular ATI is associated with pathology. The role of ATI in eliciting an immune response has been shown in animal and human research models and is considered to be an important entry antigen for celiac and non-celiac wheat sensitivity. ATI primarily triggers an innate immune response involving macrophages, neutrophils, and gut dendritic cells by activating TLR complexes. In contrast, gliadin does not appear to be the primary cause, as the typical gliadin-induced gut response profile is not found, and gliadin primarily activates adaptive immune markers (e.g., IL-6, IL-21, and INF- γ), not found in patients with non-celiac wheat sensitivity.

Patients with non-celiac wheat sensitivity show elevated expression levels of TLR2, increased numbers of intraepithelial alpha and beta lymphocytes, and decreased numbers of T regulatory cells. Also, these patients have elevated levels of Lipopolysaccharide Binding Protein (LBP) and soluble CD14 protein. In addition, these patients have elevated levels of fatty acid binding protein 2(FABP2) in their blood circulation, which is a marker of intestinal epithelial cell injury (Uhde et al, Gut 65,1930 (2016)). It has now been accepted that patients with non-celiac wheat sensitivity exhibit increased intestinal permeability, which allows antigens to pass through the lamina propria.

Changes in intestinal flora may also play a role in non-celiac wheat susceptibility. The immune markers seen in patients are primarily innate immune response markers, which provide evidence that microbiota is functioning. However, evidence for the role of microbial populations remains unclear. From a compositional standpoint, it appears that patients may have a lower abundance of butyrate-producing bacteria and bifidobacteria.

In addition, there is some evidence that the condition occurs in individuals with some genetic predisposition. The genetic susceptibility is higher than the general population but lower than patients with celiac disease (with a powerful genetic component). However, such association with genes is not clear at present.

The diagnosis of this condition is complex and many patients are reluctant to undergo this procedure. The first step in the diagnosis is to exclude celiac disease and wheat and/or gluten allergy. This was done by placing the patient on a gluten-containing diet for a period of 6 weeks. During this period, several tests were performed to rule out wheat allergy (i.e. wheat specific IgE and skin prick test) and celiac disease (i.e. IgA-tTG, IgG-DGP and IgA-EMA). If necessary, a duodenal biopsy may be performed for confirmation. The second step involves starting the patient on a gluten-free diet for a period of 6 weeks and monitoring the symptomatic response. The symptomatic response was assessed using the Gastrointestinal Symptom Rating Scale (GSRS) and the Numerical Rating Scale (NRS). If the patient does not have an improvement in symptoms within 6 weeks after the start of the diet, the diagnosis of the condition should be excluded and other diagnoses need to be explored, such as IBS and other functional bowel disorders. The third step involves reintroduction of the gluten-containing diet. In this step, patients are randomly assigned to one of two protocols. Patients received a gluten-free diet + placebo or a gluten-free diet + gluten for one week. The patient then received a strict gluten-free diet for a 1-week washout period followed by a crossover (crossover) for 1 week. Positive results were indicated by 30% improvement of symptoms after introduction of the gluten-free diet or by 30% appearance of symptoms after introduction of the gluten-containing diet. Values below 30% are considered negative.

The difficulty of diagnosis means that the prevalence of the condition is not clear. However, this condition is becoming a more common diagnosis. As a result, the prevalence of this condition is reported to vary widely from 0.6 to 6% among western populations. This lack of diagnostic ability has led patients to begin gluten-free diets after self-diagnosis without any formal clinical testing or regulatory advice by their physicians. Thus, consumption of gluten-free foods is becoming increasingly popular in the western world. A poll conducted on 7 months of 2015 for glopopu (Gallup) showed that 20% of americans chose a gluten-free diet, while 17% of americans indicated that they avoided eating gluten-free food.

Currently, there is no cure for this condition and the only accepted treatment is to place the patient on a gluten-free diet, which often helps to address intestinal and extra-intestinal symptoms. However, it is recommended to continue to consume the gluten-free diet for life later. The consequences of this nutrition are unclear, as gluten-containing grains also contain many essential nutrients; in particular fibers. In addition, the removal of gluten-containing foods can have a significant impact on the quality of life of the patient. Preferably, the patient is provided with an intervention that at least substantially alleviates the symptoms or prevents the recurrence of the condition, even when fed wheat and/or gluten.

Thus, there remains a need for methods and compounds for managing non-celiac wheat and/or gluten sensitivity in humans to allow for the consumption of wheat and/or gluten.

Disclosure of Invention

A first aspect of the present invention relates to a Human Milk Oligosaccharide (HMO) for use in:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans.

A second aspect of the invention relates to a synthetic composition for:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans,

the composition comprises at least one Human Milk Oligosaccharide (HMO).

Preferably, the synthetic composition contains HMO in an amount of 1g to 15 g; more preferably, 2g to 10 g. For example, the synthetic composition may contain from 3g to 7g of HMO.

The synthetic composition may contain bifidobacteria, such as Bifidobacterium longum (Bifidobacterium longum) and/or Bifidobacterium bifidum (Bifidobacterium bifidum).

A third aspect of the invention relates to a method for treating non-celiac wheat and/or gluten sensitivity in a human, the method comprising administering to the human an effective amount of at least one Human Milk Oligosaccharide (HMO).

A fourth aspect of the invention relates to a method for secondary prevention of non-celiac wheat and/or gluten sensitivity in a human, the method comprising administering to the human an effective amount of at least one Human Milk Oligosaccharide (HMO).

A fifth aspect of the present invention relates to a method for inducing wheat and/or gluten tolerance in a patient suffering from non-celiac wheat and/or gluten sensitivity, the method comprising administering to the patient an effective amount of at least one Human Milk Oligosaccharide (HMO).

Preferably, HMOs are administered to the patient at the same time as the gluten-containing grain is consumed.

The amount of HMO administered is preferably effective to increase the abundance of butyrate producing bacteria and/or bifidobacteria in the human intestine. Furthermore, the amount of HMO administered is preferably effective to improve the intestinal barrier properties in humans, especially in the colon.

Preferably, HMO is administered to the human in an amount of 1g to 15g per day, more preferably 2g to 10g per day. For example, 3g to 7g may be administered to a human per day. Preferably, HMOs are administered to humans for a period of at least 7 consecutive days (1 week), more preferably for at least 14 consecutive days (2 weeks).

Higher doses may be administered to patients during treatment of non-celiac wheat and/or gluten sensitivity, while lower doses are used to induce wheat and/or gluten tolerance or as secondary prevention. Preferably, HMOs are administered to humans for at least 1 week, more preferably at least 2 weeks during the treatment period. HMOs may be administered to a human for at least 4 weeks, more preferably at least 8 weeks, to induce wheat and/or gluten tolerance or as a secondary prevention. The dose administered during the treatment period is preferably from about 3g to about 15g per day (e.g., from about 4g to about 7.5g per day), and the dose administered during the wheat and/or gluten tolerance-inducing or secondary prevention period is from about 2g to about 7.5g per day (e.g., from about 2g to about 5g per day). In one embodiment, the method of the invention comprises a first stage of treatment of non-celiac wheat and/or gluten sensitivity followed by a second stage for induction of wheat and/or gluten tolerance or secondary prevention. Preferably, HMO is administered to the human for a period of at least 7 consecutive days (1 week), more preferably at least 14 consecutive days (2 weeks) in a first phase until non-celiac wheat and/or gluten sensitivity improves (typically 2-3 months), followed by a second phase to induce wheat and/or gluten tolerance or as a secondary prevention in which HMO is administered to the human for at least (consecutive) 4 weeks, more preferably at least 8 weeks.

A sixth aspect of the invention relates to an encapsulant (pack) for:

-treatment of non-celiac wheat and/or gluten sensitivity in humans,

-inducing wheat and/or gluten tolerance in patients with non-celiac wheat and/or gluten sensitivity, and/or

Secondary prevention of non-celiac wheat and/or gluten sensitivity in humans,

the package comprises an effective amount of at least one Human Milk Oligosaccharide (HMO) in at least 14 independent daily doses.

Preferably, each dose contains from about 1g to 15g of human milk oligosaccharide, more preferably from about 2g to about 10 g; for example, about 3g to about 7 g. Preferably, the package comprises at least 21 daily doses, more preferably at least 28 daily doses; for example, at least 35 daily doses. The package may include instructions for use.

The seventh aspect of the present invention is:

-one or more Human Milk Oligosaccharides (HMOs),

-a synthetic composition comprising one or more Human Milk Oligosaccharides (HMOs), or

-a package comprising an effective amount of at least one or more human milk oligosaccharides for at least 14 independent daily doses

Use in the dietary management of patients suffering from non-celiac wheat and/or gluten sensitivity.

In certain embodiments of any aspect, the HMO may be a neutral HMO or an acidic HMO. The neutral HMOs may be one or more fucosylated HMOs or one or more nonfucosylated HMOs. Preferably, the HMO is selected from 2 ' -FL, 3-FL, DFL, LNT, LNnT, 3 ' -SL, 6 ' -SL, LNFP-I or mixtures thereof. Preferably, the HMO comprises, consists of, or consists essentially of: 2' -FL and at least one of LNnT and LNT; at least one of 2 '-FL and DFL and at least one of LNnT and LNT (e.g., at least one of 2' -FL, DFL, and LNnT and LNT); 2 '-FL and 6' -SL; DFL and 6' -SL; 2 '-FL, DFL and 6' -SL; 2 '-FL, 6' -SL and at least one of LNnT and LNT; 2 '-FL, DFL, 6' -SL, and at least one of LNnT and/or LNT.

Detailed Description

It has now surprisingly been found that oral or enteral administration of one or more Human Milk Oligosaccharides (HMOs) to patients suffering from non-celiac wheat and/or gluten sensitivity may alleviate the symptoms of this condition. Furthermore, HMOs surprisingly induce tolerance to wheat and/or gluten in patients with non-celiac wheat and/or gluten sensitivity; the patient is allowed to eat wheat and/or gluten with reduced or no symptoms. Thus, human milk oligosaccharides may be used as a dietary secondary prevention of non-celiac wheat and/or gluten sensitivity. HMOs also preferentially increase the abundance of bifidobacteria in the gastrointestinal tract, in particular bifidobacteria of the phylogenetic group Bifidobacterium adolescentis (b.adolescentis), Bifidobacterium longum (Bifidobacterium longum) and/or Bifidobacterium bifidum (Bifidobacterium bifidum). These bacteria produce lactate and acetate which in turn can be converted to butyrate by the butyrate producing bacteria.

Human milk oligosaccharides are a heterogeneous mixture of soluble glycans found in human milk. They are the third most abundant solid component next to lactose and lipids in Human milk and are present at concentrations of 5-25g/l (Bode: Human milligosaccharaides and the hair cosmetic effects, see: Handbook of dietetic and nutritional aspects of Human breath mill (Zibadi et al), pp.515-31, Wahering Academic publications (2013)). HMOs are resistant to enzymatic hydrolysis in the small intestine and are therefore largely undigested and absorbed and reach the colon intact. Most HMOs that reach the colon serve as substrates to shape the intestinal ecosystem by selectively stimulating the growth of specific bacteria. It is believed that HMOs can substantially regulate infant intestinal flora and play a decisive role in the microbiota difference between formula-fed and breast-fed infants. These differences include the predominance of bifidobacteria in the gut of breast-fed infants compared to the more diverse gut flora in formula-fed infants. This is considered beneficial for infants, as strains of bifidobacterium are considered to have a positive effect on intestinal health.

HMOs also preferentially increase the abundance of bifidobacteria in the gastrointestinal tract, in particular bifidobacteria of the phylogenetic group Bifidobacterium adolescentis (b.adolescentis), Bifidobacterium longum (Bifidobacterium longum) and/or Bifidobacterium bifidum (Bifidobacterium bifidum).

In the present specification, the following terms have the following meanings:

by "non-celiac wheat sensitivity" is meant a syndrome characterized by intestinal and extra-intestinal symptoms associated with ingestion of gluten-containing foods in subjects unaffected by celiac disease or wheat allergy. "non-celiac gluten sensitivity" has the same meaning, and the two terms may be used interchangeably. Gluten-containing foods typically include gluten-containing grains such as wheat, barley, and rye.

By "non-infant human" or "non-infant" is meant a human that is 3 years of age and older. The non-infant human may be a child, adolescent, adult or elderly human.

"human milk oligosaccharides" or "HMOs" mean the complex carbohydrates found in human breast milk (Urshima et al: MillkOligosaccharides, Novascience publishers (2011)); chenaddv. carbohydr. chem. biochem.72,113 (2015)). HMOs have a core structure comprising a lactose unit at the reducing end, which may be extended by one or more β -N-acetyl-lactosaminyl and/or one or more β -lacto-N-disaccharide (β -lacto-N-biosyl) units, and which may be substituted by α L-fucopyranosyl (fucopyranosyl) and/or α -N-acetyl-neuraminic acid (sialyl) moieties. In this regard, the non-acidic (or neutral) HMOs do not contain sialic acid residues, whereas acidic HMOs have at least one sialic acid residue in their structure. The non-acidic (or neutral) HMOs may be fucosylated or non-fucosylated. Examples of such neutral non-fucosylated HMOs include lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-neohexose (LNnH), p-lacto-N-neohexose (pLNnH), p-lacto-N-hexose (pLNH), and lacto-N-hexose (LNH). Examples of neutral fucosylated HMOs include 2 '-fucosyllactose (2' -FL), lacto-N-fucopentaose I (LNFP-I), lacto-N-difucohexaose I (LNDFH-I), 3-fucosyllactose (3-FL), Difucosyllactose (DFL), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP-III), lacto-N-difucohexaose III (LNDFH-III), fucosyl-lacto-N-hexaose II (FLNH-II), lacto-N-fucopentaose V (LNFP-V), lacto-N-fucopentaose VI (LN-FP), lacto-N-difucohexaose II (LNDFH-II), fucosyl-lacto-N-hexose I (FLNH-I), fucosyl-p-lacto-N-hexose I (FpLNH-I), fucosyl-p-lacto-N-neohexose ii (fplnnh ii), and fucosyl-lacto-N-neohexose (flnhh). Examples of acidic HMOs include 3 ' -sialyllactose (3 ' -SL), 6 ' -sialyllactose (6 ' -SL), 3-fucosyl-3 ' -sialyllactose (FSL), LST a, fucosyl-LST a (FLST a), LST b, fucosyl-LST b (FLSTb), LST c, fucosyl-LST c (FLST c), sialyl-lnh (SLNH), sialyl-lacto-N-hexose (SLNH), sialyl-lacto-N-neohexose I (SLNH-1), sialyl-lacto-N-neohexose II (SLNH-II), and disialyl-lacto-N-tetraose (DSLNT).

"synthetic composition" means a composition that is artificially prepared, and preferably means a composition that contains at least one compound that is produced (e.g., by a chemical reaction, enzymatic reaction, or recombination) chemically and/or biologically ex vivo. In some embodiments, the synthetic composition of the invention may be the same as the naturally occurring composition, but preferably is different. Synthetic compositions typically comprise one or more compounds capable of reducing the symptoms of or inducing tolerance to wheat and/or gluten sensitivity in humans, including one or more HMOs. Also, in some embodiments, the synthetic compositions may comprise one or more nutritional or pharmaceutically active components that do not adversely affect the efficacy of the above-described compounds. Some non-limiting embodiments of the synthetic compositions of the present invention are also described below.

"microbial population", "microflora" and "microbiome" refer to a population of living microorganisms that inhabit body organs or parts in general, and gastrointestinal organs in particular in humans. The most major members of the gastrointestinal microbiota include microorganisms of the phyla Firmicutes, Bacteroidetes, actinomycetes, Proteobacteria, syntrophic bacteria, Verrucomicrobia, Fusobacteria and eurycota. Microorganisms at the genus level include Bacteroides (Bacteroides), Faecalibacterium (Faecalibacterium), Bifidobacterium (Bifidobacterium), Roseburia (Roseburia), Alisipes, Coriolis (Collinsella), Blautia (Blautia), enterococcus (Coprococcus), Ruminococcus (Ruminococcus), Eubacterium (Eubacterium), and Dorea; microorganisms at the species level include Bacteroides monomorphus (Bacteroides), Alisiples vitronensis, Parabateria merdae, Ruminococcus braunii, Dorea longticana, Bacteroides coprinus (Bacteroides caccae), Bacteroides thetaiotaomicron (Bacteroides thetaiotaomicron), Eubacterium halobacterium (Eubacterium halii), Ruminococcus torvus (Ruminococcus torqueus), Clostridium coprinus (Faecalixima), Ruminococcus acidilactici (Ruminococcus acidilasus), Coprinus aerogenes (Colinobacillus aeolicus), Doreafiiginens, Bacteroides vulus vulgatoides (Bacteroides vulus vygaeus) and Roseburia terreus (Rosebuiella terrestris). The gastrointestinal microbiota includes mucosa-associated microbiota located in or attached to the mucus layer covering the epithelium of the gastrointestinal tract, as well as lumen-associated microbiota found in the lumen of the gastrointestinal tract.

By "enteral administration" is meant any conventional form for delivering the composition to a non-infant which causes deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include oral, sublingual and rectal feeding through nasogastric or jejunal tubes.

By "oral administration" is meant any conventional form for delivering a composition to a human through the mouth. Thus, oral administration is one form of enteral administration.

By "effective amount" is meant an amount of the composition that provides HMOs in a sufficient amount to elicit the desired therapeutic result in a human. An effective amount may be administered in one or more doses to achieve the desired therapeutic result.

By "relative abundance of bifidobacteria" is meant the abundance of bifidobacteria species relative to other bifidobacteria in the human gastrointestinal microbiota.

By "relative growth of bifidobacteria" is meant the growth of bifidobacteria species relative to other bifidobacteria in the human gastrointestinal microbiota.

"Bifidobacterium adolescentis phylogenetic group" means a bacterium selected from the group consisting of: bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium angulus (Bifidobacterium angulus), Bifidobacterium catenulatum (Bifidobacterium catenulatum), Bifidobacterium pseudocatenulatum (Bifidobacterium pseudocatenulatum), Bifidobacterium kashiwanogens, Bifidobacterium odonta (Bifidobacterium endohedral), Bifidobacterium stercoris (Bifidobacterium et al, applied. environ. Microbiol.79,336(2013), Bacillus Cell factor. 13: S4 (2014)). Preferably, the bifidobacteria of the phylogenetic group of bifidobacterium adolescentis are bifidobacterium adolescentis and/or bifidobacterium pseudocatenulatum.

By "treatment" is meant the resolution of a medical condition or disease with the aim of improving or stabilizing the prognosis or potential nutritional needs of the person being treated. Thus, treatment includes dietary or nutritional management of medical conditions or diseases by addressing the nutritional needs of the person being treated. "treatment" and "treatment" have grammatically corresponding meanings.

By "dietary management" is meant all or part of a meal for a patient who has experienced the following due to a disease, disorder or medical condition:

the ability to ingest, digest, absorb, metabolize or excrete normal food or certain nutrients or metabolites contained therein is limited, impaired or disturbed, or

-having other medically defined nutrient requirements (see: Committee bulletin of the Committee of the European Committee for the classification of food for special medical use, European official gazette C401, 25.11.2017, pages 10-11).

By "modulation of the microbiota" is meant an effect that exerts a modulation or control over the microbiota, such as an effect that results in an increase in the intrinsic intestinal abundance of bifidobacteria (bifidobacteria), Barnesiella, faecalis (Faecalibacterium) and/or other butyrate producing bacteria. In another example, the effect may cause a decrease in the intestinal abundance of active ruminococcus (ruminococcus gnavus) and/or proteus. "Proteobacteria" is a gram-negative phylum and includes various pathogenic bacteria such as Escherichia (Escherichia), Salmonella (Salmonella), Vibrio (Vibrio), Helicobacter (Helicobacter), Yersinia (Yersinia) and many other well-known genera.

"treatment" means treatment or action taken to reduce or eliminate the symptoms of a disease or pathological condition.

As used herein, "prophylactic treatment" or "prevention" refers to treatment or action taken to reduce the risk of onset or recurrence of a disease.

"Secondary prevention" refers to the prevention of the onset of a condition in a high risk patient, or the prevention of recurrence of symptoms in a patient already suffering from the condition. A "high risk" patient refers to an individual who is susceptible to disease, e.g., a person with a family history of the condition.

HMOs can be isolated or enriched from one or more milks secreted from mammals (including but not limited to human, bovine, ovine, porcine, or caprine species) by well-known methods. HMOs can also be produced by well-known methods using microbial fermentation, enzymatic methods, chemical synthesis, or a combination of these techniques. As examples, using chemical methods, LNnT can be made as described in WO 2011/100980 and WO2013/044928, LNT can be synthesized as described in WO 2012/155916 and WO2013/044928, a mixture of LNT and LNnT can be made as described in WO 2013/091660, 2 '-FL can be made as described in WO 2010/115934 and WO2010/115935, 3-FL can be made as described in WO 2013/139344, 6' -SL and salts thereof can be made as described in WO 2010/100979, sialylated oligosaccharides can be made as described in WO 2012/113404, and a mixture of human milk oligosaccharides can be made as described in WO 2012/113405. As an example of enzymatic production, sialylated oligosaccharides may be manufactured as described in WO 2012/007588, fucosylated oligosaccharides may be manufactured as described in WO 2012/127410, and various mixtures of human milk oligosaccharides may be advantageously prepared as described in WO2012/156897 and WO 2012/156898. Biotechnological methods describing how to make core (nonfucosylated neutral) human milk oligosaccharides (optionally substituted with fucose or sialic acid) using genetically modified escherichia coli can be found in WO 01/04341 and WO 2007/101862.

In any of the above aspects, the HMO may be a single HMO or any mixture of HMOs suitable for the purposes of the present invention. The HMO may be a neutral HMO or an acidic HMO. In one embodiment, the neutral HMOs are one or more fucosylated HMOs; in another embodiment, the neutral HMOs are one or more nonfucosylated HMOs. In particular, the fucosylated neutral HMOs are selected from the list consisting of: 2 '-FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, LNFP-VI, LNDFH-I, LNDFH-II, LNDFH-III, FLNH-I, FLNH-II, FLNnH, FpLNH-I and F-pLNnH II, preferably 2' -FL, and the nonfucosylated neutral HMO is selected from the list consisting of LNT, LNnT, LNH, LNnH, pLNH and pLNnH, e.g., LNnT. The one or more fucosylated HMOs may be, for example, a mixture comprising, consisting of, or consisting essentially of 2' -FL and DFL.

In one embodiment, the mixture comprises, consists of, or consists essentially of: neutral HMOs, preferably at least a first neutral HMO and at least a second neutral HMO, wherein the first neutral HMO is a fucosylated neutral HMO and the second neutral HMO is a non-fucosylated neutral HMO. The fucosylated neutral HMOs and nonfucosylated HMOs may be present in a mass ratio of about 4:1 to 1: 1. In particular, the mixture of HMOs comprises, consists of, or consists essentially of fucosylated HMOs selected from the list consisting of 2' -FL, 3-FL, DFL, LNFP-I, LNFP-II, LNFP-III, LNFP-V, LNDFH-I, LNDFH-II, LNDFH-III, FLNH-I, FLNH-II, flnhh, FpLNH-I and F-pLNnHII, as well as non-fucosylated neutral HMOs selected from the list consisting of LNT, LNnT, LNH, LNnH, pLNH and plnhh. More preferably, the mixture of neutral HMOs contains, consists of, or consists essentially of: a fucosylated HMO selected from the list consisting of 2' -FL, 3-FL and DFL, and a non-fucosylated neutral HMO selected from the list consisting of LNT and LNnT; advantageously, the mixture comprises, consists of, or consists essentially of: 2' -FL and at least one of LNnT and LNT; or at least one of 2' -FL and DFL, and at least one of LNnT and LNT; or 2' -FL, DFL and at least one of LNnT and LNT.

In other embodiments, the mixture comprises, consists of, or consists essentially of at least a first (acidic) HMO and at least a second (neutral) HMO, wherein the first (acidic) HMO is selected from the list consisting of 3 ' -SL, 6 ' -SL, and FSL and the second (neutral) HMO is selected from the list consisting of 2 ' -FL, 3-FL, DFL, LNT, and LNnT; advantageously, the mixture comprises, consists or consists essentially of: 2 '-FL and 6' -SL; or 6 '-SL and at least one of 2' -FL and DFL; or 2 '-FL, 6' -SL and at least one of LNnT and LNT; or 2 '-FL, DFL, 6' -SL and at least one of LNnT and/or LNT.

Furthermore, in one embodiment, the synthetic composition may be in the form of a nutritional composition. For example, the nutritional composition may be a food composition, a nutritional supplement, a medical food or a food for a specific medical purpose, a nutritional supplement, or the like. The nutritional composition may contain a source of protein, lipid and/or digestible carbohydrate and may be in powder or liquid form. The composition may be designed as the sole source of nutrition or as a nutritional supplement.

Suitable protein sources include milk protein, soy protein, rice protein, pea protein and oat protein, or mixtures thereof. The milk protein may be in the form of a milk protein concentrate, milk protein isolate, whey protein or casein or a mixture of both. The protein may be intact or hydrolysed, partially hydrolysed or extensively hydrolysed. Hydrolyzed protein offers the advantage of being easily digestible, which is of great importance for people with inflamed or damaged gastrointestinal tracts. Proteins may also be provided in the form of free amino acids. Protein may comprise from about 5% to about 30%, typically from about 10% to 20% of the energy of the nutritional composition.

The protein source may be the following: glutamine, threonine, cysteine, serine, proline, or combinations of these amino acids. The glutamine source may be glutamine dipeptide and/or a glutamine rich protein. Since the intestinal cells use glutamine as an energy source, glutamine can be included. Threonine, serine and proline are important amino acids for the production of mucins. Mucins coat the gastrointestinal tract and can improve intestinal barrier function and mucosal healing. Cysteine is the main precursor of glutathione, which is the key to the body's antioxidant defense.

Suitable digestible carbohydrates include maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup solids, high fructose corn syrup, rice derived carbohydrates, pea derived carbohydrates, potato derived carbohydrates, tapioca starch, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), or mixtures thereof. Preferably, the composition is reduced or free of added lactose or other FODMAP carbohydrates. Generally, the digestible carbohydrate provides about 35% to about 55% of the energy of the nutritional composition. A particularly suitable digestible carbohydrate is low Dextrose Equivalent (DE) maltodextrin.

Suitable lipids include Medium Chain Triglycerides (MCT) and Long Chain Triglycerides (LCT). Preferably, the lipid is a mixture of MCT and LCT. For example, MCTs can comprise from about 30% to about 70% by weight of the lipid, more specifically from about 50% to about 60% by weight. MCT have the advantage of being easily digestible, which is important for people with inflamed or damaged gastrointestinal tract. Typically, the lipids provide from about 35% to about 50% of the energy of the nutritional composition. Lipids may contain essential fatty acids (omega-3 and omega-6 fatty acids). Preferably, these polyunsaturated fatty acids provide less than about 30% of the total energy of the lipid source.

Suitable sources of long chain triglycerides are rapeseed oil, sunflower oil, palm oil, soybean oil, milk fat, corn oil, high oil oils and soy 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 omega-6 (n-6) to omega-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 from about 6:1 to about 9: 1.

The nutritional composition may further comprise vitamins and minerals. If the nutritional composition is intended as the sole source of nutrition, it preferably includes a complete vitamin and mineral profile. Examples of vitamins include vitamin A, B-complex (e.g., 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.

The nutritional composition may also comprise carotenoids such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoid included may range from about 0.001 μ g/ml to about 10 μ g/ml. Lutein may be included in amounts of about 0.001 to about 10 μ g/ml, preferably about 0.044 to about 5 μ g/ml lutein. Lycopene may be included in an amount of from about 0.001 μ g/ml to about 10 μ g/ml, preferably from about 0.0185 μ g/ml to about 5 μ g/ml of lycopene. The beta-carotene may comprise from about 0.001 to about 10mg/ml, for example from about 0.034 to about 5 μ g/ml of beta-carotene.

The nutritional composition preferably also contains a low concentration of sodium; for example, from about 300mg/l to about 400 mg/l. The concentration of residual electrolytes may be desirable without unnecessarily burdening renal function with renal solutes. For example, potassium is preferably present in the range of about 1180mg/l to about 1300 mg/l; and chloride is preferably present in the range of about 680mg/l to about 800 mg/l.

The nutritional composition may further comprise various other conventional ingredients such as preservatives, emulsifiers, thickeners, buffers, fibers and prebiotics (e.g. fructooligosaccharides, galactooligosaccharides), probiotics (e.g. bifidobacterium animalis (b.animalis) subsp. lactis) BB-12, bifidobacterium lactis (b.lactis) HN019, bifidobacterium lactis Bi07, bifidobacterium infantis (b.infarnatis) ATCC 15697, lactobacillus rhamnosus (l.rhamnosus) GG, lactobacillus rhamnosus HNOOl, lactobacillus acidophilus (l.acidophilus) LA-5, lactobacillus acidophilus NCFM, lactobacillus fermentum (l.fermentum) CECT5716, bifidobacterium longum (b.longum) BB536, bifidobacterium longum AH1205, bifidobacterium longum 1206, bifidobacterium breve (b.breve) M-16V, lactobacillus reuteri (l.reuteri) ATCC 30, lactobacillus reuteri ATCC 55785, anti-inflammatory lactobacillus strain ATCC 6485, lactobacillus plantarum ATCC 938, and anti-inflammatory compounds including lactobacillus plantarum DSM 6485, lactobacillus plantarum, Carotenoids, ascorbic acid/vitamin C, ascorbyl palmitate, polyphenols, glutathione and superoxide dismutase (cantaloupe), other bioactive factors (e.g. growth hormone, cytokines, TFG- β), colorants, flavors and stabilizers, lubricants and the like.

The nutritional compositions may be formulated as soluble powders, liquid concentrates, or ready-to-use formulations. The composition may be administered to a person in need thereof via nasogastric tube or orally. Various flavoring agents, fibers, and other additives may also be present.

The nutritional compositions may be prepared by any of the usual manufacturing techniques for preparing nutritional compositions in solid or liquid form. For example, the composition may be prepared by combining the various feed solutions. A protein-in-fat feed solution may be prepared by heating and mixing a lipid source, and then adding an emulsifier (e.g., lecithin), a fat-soluble vitamin, and at least a portion of the protein source while heating and stirring. The carbohydrate feed solution is then prepared by adding minerals, trace and ultra trace minerals, thickeners or suspending agents to water while heating and stirring. The resulting solution was held under constant heating and stirring for 10 minutes prior to adding the carbohydrates (e.g., HMOs and digestible carbohydrate source). The resulting feed solution is then blended together while heating and stirring and the pH is adjusted to 6.6-7.0, and the composition is then subjected to high temperature short time processing during which the composition is heat treated, emulsified and homogenized, and then allowed to cool. Water soluble vitamins and ascorbic acid are added, the pH is adjusted to the desired range if necessary, flavoring agents are added, and water is added to achieve the desired total solids content.

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

When the nutritional product is a ready-to-eat nutritional liquid, the total concentration of HMOs in the liquid is preferably from about 0.1% to about 1.5%, including from about 0.2% to about 1.0%, for example from about 0.3% to about 0.7%, by weight of the liquid. When the nutritional product is a concentrated nutritional liquid, the total concentration of HMOs in the liquid is preferably from about 0.2% to about 3.0%, including from about 0.4% to about 2.0%, for example from about 0.6% to about 1.5%, by weight of the liquid.

In another embodiment, the nutritional composition is in unit dosage form. The unit dosage form may contain an acceptable food grade carrier, for example, a phosphate buffered saline solution, a mixture of ethanol in water, and emulsions such as an oil/water or water/oil emulsion, and various wetting agents or excipients. The unit dosage form may also contain other materials which do not produce a deleterious, allergic, or other untoward effect when administered to a human. Carriers and other materials may include solvents, dispersants, coatings, absorption promoters, controlled release agents and one or more inert excipients, such as starches, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders and disintegrating agents.

The unit dosage forms of the invention may be administered orally, for example as tablets, capsules or pills containing a predetermined amount of the mixture, or as powders or granules containing a mixture of predetermined concentrations, or gels, pastes, solutions, suspensions, emulsions, syrups, boluses, lozenges or syrups containing a predetermined concentration of the mixture in an aqueous or non-aqueous liquid. Compositions for oral administration may comprise one or more binders, lubricants, inert diluents, flavoring agents, and humectants. The orally administered compositions, such as tablets, may optionally be coated and may be formulated to provide sustained, delayed or controlled release of HMOs.

The unit dosage forms of the invention may also be administered via nasogastric tube or infused directly into the gastrointestinal tract or stomach.

The unit dosage forms of the present invention may also include therapeutic agents such as antibiotics, probiotics, analgesics, and anti-inflammatory agents. Suitable dosages of such compositions for use in humans can be determined in a conventional manner based on factors such as the condition, immune status, weight and age of the human. In some cases, the concentration of the dose will be similar to that found for HMO in human breast milk compositions. The desired amount is generally from about 1g to about 15g per day, in certain embodiments from about 2g to about 10g per day, for example from about 3g to about 7g per day. Appropriate dosage regimens can be determined by methods known to those skilled in the art.

In another embodiment, the HMO may be formulated as a pharmaceutical composition. The pharmaceutical compositions may contain a pharmaceutically acceptable carrier, for example, a phosphate buffered saline solution, a mixture of ethanol in water, water and an emulsion (e.g., an oil/water or water/oil emulsion), and various wetting agents or excipients. The pharmaceutical composition may also contain other substances which do not produce harmful, allergic or other unwanted reactions when administered to a non-infant. Carriers and other materials may include solvents, dispersants, coatings, absorption promoters, controlled release agents and one or more inert excipients, such as starches, polyols, granulating agents, microcrystalline cellulose, diluents, lubricants, binders and disintegrating agents.

The pharmaceutical compositions may be administered orally, for example as a powder or granules containing a predetermined amount of tablets, capsules or pills, or as a powder or granules containing a predetermined concentration, or as a gel, paste, solution, suspension, emulsion, syrup, bolus, electuary or syrup in an aqueous or non-aqueous liquid containing a predetermined concentration. Compositions for oral administration may comprise binders, lubricants, inert diluents, flavoring agents and wetting agents. Compositions for oral administration, such as tablets, may optionally be coated and may be formulated to provide sustained, delayed or controlled release of the mixture therein.

The pharmaceutical compositions may also be administered by rectal suppository, aerosol tube, nasogastric tube, or direct infusion into the gastrointestinal tract or stomach.

The pharmaceutical composition may also include therapeutic agents such as antibiotics, probiotics, analgesics, and anti-inflammatory agents. Suitable dosages of these compositions for use in humans can be determined in a conventional manner based on factors such as condition, immune status, weight and age. In some cases, the concentration of the dose will be similar to the concentration of HMO in human breast milk. The desired amount generally ranges from about 1g to about 15g per day, in certain embodiments from about 2g to about 10g per day, for example from about 3g to about 7g per day. Appropriate dosage regimens can be determined by conventional methods.

For the treatment of non-celiac wheat and/or gluten sensitivity in humans, the amount of HMO that needs to be administered will vary depending on factors such as the risk and severity of fatigue, any potential medical condition or disease, age, form of the composition and other drugs being administered. In addition, the amount may vary depending on whether HMO is used to reduce the symptoms of activity (when the dose may be higher) or whether HMO is used to induce tolerance and/or as a secondary prophylaxis (when the dose may be lower). However, the desired amount can be readily set by the physician, and is typically in the range of about 1g to about 15g per day, in certain embodiments about 2g to about 10g per day, for example about 3g to about 7g per day. Suitable dosages may be determined based on several factors including, for example, body weight and/or condition, severity of non-celiac wheat and/or gluten sensitivity being treated or prevented, other conditions and/or diseases, incidence and/or severity of side effects, and mode of administration. Appropriate dosage ranges can be determined by methods known to those skilled in the art. For example, for treating non-celiac wheat and/or gluten sensitivity in a human, the dose may be from about 3g to about 15g per day, preferably from 4g to 10g per day, while for inducing wheat and/or gluten tolerance or in secondary prevention, the dose may be from about 2g to about 7.5g per day, preferably from 2g to 5g per day. In a combination treatment regimen, the dose may be higher (e.g., 3g to 15g per day, preferably 4g to 7.5g) during an initial treatment phase (first phase) for treating non-celiac wheat and/or gluten sensitivity, followed by a phase (second phase) that induces wheat and/or gluten tolerance or secondary prevention, and the dose may be reduced (e.g., 2g to 7.5g per day, preferably 2g to 5g per day).