Compositions of galactooligosaccharides and uses thereof

Abstract

A prebiotic oligosaccharide composition for use in the prevention, amelioration or treatment of upper respiratory system (URS) and upper gastrointestinal tract (GI) conditions and / or diseases. The present invention relates to a composition comprising an oligosaccharide compound, such as a galactooligosaccharide compound, comprising, based on the total weight of oligosaccharide compounds present in the composition: (a) at least 8 wt% of Gal-(beta1-3)-Gal-(beta1-4)-Xa; (b) at least 3 wt% of Gal-([beta] 1-3)-Gal-([beta] 1-3)-Xb; and (c) at least 5 wt% of Gal-(beta1-3)-Gal-(beta1-2)-Xc; wherein Xa, Xb and Xc are independently selected from monosaccharides. These compositions contain relatively high levels of oligosaccharide compounds (a), (b) and (c) and relatively high levels of [beta] 1-3Gal-Gal linkages compared to known oligosaccharide compositions. The invention also relates to a composition comprising an oligosaccharide compound for improving athlete performance. These particular characteristics of the composition are believed to provide benefits for the eater's intestinal health, such as reducing the symptoms of URS and upper GI conditions and / or diseases. A method of preparing the composition and the use of the composition as a dietary supplement or medicament are also disclosed.

Description

Technical Field

[0001] The present invention relates to a composition comprising an oligosaccharide compound for preventing, improving or treating upper respiratory system (URS) and gastrointestinal tract (GI) disorders and / or diseases. The present invention also relates to a composition comprising an oligosaccharide compound for improving athlete performance. The present invention also relates to a method for preparing the composition and a formulation of the composition as a dietary supplement and / or medicament. Specifically, the present invention relates to an oligosaccharide composition having a relatively high content of certain beneficial oligosaccharides. Background Art

[0002] The human intestinal microbiota (or microbial flora) includes pathogenic, benign and beneficial microbial genera. The dominance of the former may lead to acute (such as gastroenteritis) and chronic (such as inflammatory bowel disease, irritable bowel syndrome and some intestinal cancers) intestinal diseases. The human colon microbiota is acquired at birth. Bifidobacteria are dominant in breastfed infants and can easily outcompete other types of bacteria. This is because breast milk components stimulate the growth of bifidobacteria. However, many factors can affect the composition and activity of the intestinal microbiota throughout an individual's life, such as the environment, drug treatment and diet, especially the consumption of highly processed foods (see "Role of the gut microbiota in nutrition and health", Valdes, AM et al., BMJ, 2018, 361).

[0003] Certain components of the gut microbiota have been implicated in the etiology of intestinal diseases. For example, mycobacteria have been implicated in Crohn's disease; ulcerative colitis may be triggered by sulfate-reducing bacteria, and there may be bacteria implicated in the development of intestinal cancer. It would be beneficial if the selective growth of inherent beneficial gut bacteria could be promoted by the ingestion of prebiotics. A prebiotic is a substrate that is selectively utilized by the host microbiota, which has a health benefit. The use of such a prebiotic may have a sustained beneficial effect of inhibiting the growth of pathogenic microbiota.

[0004] There have been attempts to affect the balance of intestinal microbiota by adding one or more of these microbial strains to a suitable food carrier, in order to favor beneficial microorganisms (such as bifidobacteria), with the purpose of conferring health benefits to the host. These live microorganisms supply additives and are called probiotics. However, it is difficult to ensure the survival of live bacteria in food after digestion, so the actual effect of probiotics may be very limited. Therefore, prebiotics may be a more promising option for having a positive impact on the intestinal microbiota of patients (see Walter J, Maldonado-Gómez MX, Martínez I. "To engraft or not to engraft: an ecological framework for gut microbiome modulation with livemicrobes", Curr Opin Biotechnol. 2018, 49, 129-139).

[0005] As mentioned above, an alternative to dietary manipulation of the gut microbiota is the use of prebiotics, which are defined as non-digestible food ingredients that beneficially affect the subject by selectively stimulating the growth and / or activity of one or a limited number of bacteria in the intestine, thereby improving the health of the host.

[0006] Upper gastrointestinal (GI) disorders and / or diseases affect many people around the world, resulting in recurring and uncomfortable symptoms. For example, upper gastrointestinal disorders and / or diseases may include heartburn, regurgitation, gastroesophageal reflux disease (GERD), gastritis, and dyspepsia.

[0007] Many drugs have been developed to treat upper gastrointestinal disorders, although none are without disadvantages.

[0008] Treatment of upper gastrointestinal disorders may include alkaline agents and stomach acid suppressants and / or adjustments to certain foods in the individual's diet. Antacids (such as aluminum hydroxide and magnesium hydroxide) are commonly used to neutralize stomach acid, resulting in an increase in gastric and duodenal pH. Antacids act quickly but only last for a short time and do not always prevent heartburn when taken before foods or drinks that may trigger symptoms.

[0009] H2-receptor antagonists (also called H2-blockers) are prescription drugs that are also used to treat upper gastrointestinal tract disorders by inhibiting the action of histamine on stomach parietal cells, thereby inhibiting acid secretion. Examples of H2 receptor antagonists include ranitidine hydrochloride, cimetidine, nizatidine, rabeprazole, and famotidine. Although H2 receptor antagonists are effective, there is a delay between taking them and symptom relief, and the drugs are expensive.

[0010] Proton pump inhibitors ("PPIs") are another alternative medication for treating upper gastrointestinal disorders. PPIs are prescription medications that target the amount of gastric acid produced by the stomach's acid-producing cells. Medications include omeprazole, omeprazole IR, pantoprazole, lansoprazole, and esomeprazole. PPIs show a delay between taking and symptom relief and are expensive.

[0011] Respiratory disorders and / or illnesses typically include cough, sneezing, headache, congestion, sore throat, stuffy nose, rhinitis, and fever, etc. Medication includes liquid elixirs, cough syrups, cold and flu capsules, cold and flu tablets, allergy tablets, effervescent tablets, oral and nasal sprays, cough drops, etc.

[0012] The most commonly used drugs for the treatment of respiratory disorders and / or diseases are taken orally to solve and / or treat the onset of respiratory symptoms. These products usually contain one or more active substances dissolved or dispersed in a carrier system for ingestion or oral delivery to the bloodstream. Typically, respiratory drugs are in the form of cough drops, liquids or capsules, and respiratory products are in the form of powders and effervescent tablets. Although most drugs are relatively cheap, their effects only last for a limited time. Some treatments use a combination of antiviral and anti-inflammatory compounds. Similarly, the duration of action of these drugs is limited, and the addition of antiviral agents greatly increases the cost of these treatment types.

[0013] Elite athletes, such as rugby league players, follow physically and psychologically demanding training schedules, with frequent matches, limited recovery, and regular international travel. Collectively, these stressors may compromise immunity and increase the risk of acute upper respiratory symptoms (URS) (e.g., cough, sneezing, sore throat, and nasal congestion) and gastrointestinal symptoms (GIS) (e.g., bloating, burping, flatulence, nausea, and diarrhea) (Peters & Bateman, 1983; Drew et al., 2017; Hellard et al., 2015; Svendsen et al., 2016; Wentz et al., 2018). On average, elite rugby league players experience four episodes of upper respiratory illness and one episode of GI discomfort per season (Cunniffe et al., 2009), with the highest incidence reported in the pre-season and winter (Cunniffe et al., 2009; Tiernan et al., 2020; Keaney et al., 2021). Therefore, identifying ways to reduce these illnesses, or speed up recovery to allow return to play, is imperative for player health and team performance. The profile, genetic material, and functional activity of the gut microbial community (gut microbiome) have a significant impact on systemic immune function (Roberfoid et al., 2010). The gut microbiome can be manipulated through dietary interventions, most commonly through the consumption of probiotic or prebiotic dietary supplements. Previous studies using single or multiple probiotics have reported a reduction in the incidence of URS in active runners (Cox et al., 2007; Gleeson et al., 2011; Strasser et al., 2016). In addition, a 2015 Cochrane review reported that probiotics reduce the number and duration of URS episodes in adults and children (Hao et al., 2015). A multi-strain probiotic reduced the incidence and duration of URS in elite rugby league players by approximately 2 days (Haywood et al., 2013). However, the various probiotic strains used in different studies have created uncertainty about which strains are most beneficial for athletes’ health.

[0014] Previous studies have also shown that GOS can reduce the number of URS days and alleviate the severity of GIS in student populations (Hughes et al., 2011) and patients with travelers’ diarrhea (Drakoularakou et al., 2010; Hasle et al., 2017).

[0015] One group of compounds that are classified as prebiotics are galacto-oligosaccharides, which are galactose-containing oligosaccharides produced from lactose by the transgalactosidase activity of β-galactosidase. Known galacto-oligosaccharide prebiotic products contain a mixture of various oligosaccharide compounds in varying proportions, not all of which have the desired beneficial effects on the consumer's gut microbiota. Although these products have shown some beneficial results in improving patient health, there remains a need for further improvements in oligosaccharide compositions to fully realize the potential benefits of these prebiotics for consumers and to provide durable and inexpensive treatments for URS and GI conditions and / or diseases and / or to enhance performance in athletes. Summary of the invention

[0016] One of the objects of the present invention is to provide a composition comprising an oligosaccharide compound which solves at least one disadvantage of the prior art, whether pointed out herein or elsewhere, or provides an alternative to existing compositions. For example, an object of the present invention may be to provide a composition comprising an oligosaccharide compound which comprises a higher proportion of specific beneficial oligosaccharides than known oligosaccharide compositions.

[0017] According to some aspects of the invention, there are provided compositions, methods and uses as described in the accompanying claims. Other features of the invention will become apparent from the dependent claims and the following description.

[0018] According to a first aspect of the present invention, there is provided a composition comprising an oligosaccharide compound for preventing, improving or treating upper respiratory system (URS) and gastrointestinal (GI) disorders and / or diseases, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compound present in the composition:

[0019] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0020] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0021] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0022] Where X a , X b and X c Each is independently selected from monosaccharides.

[0023] In an embodiment related to the first aspect of the present invention, a method for preventing, ameliorating or treating URS and GI disorders and / or diseases is also provided, the method comprising administering to a subject in need of such prevention, improvement or treatment a composition comprising an oligosaccharide compound, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compound present in the composition:

[0024] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0025] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0026] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0027] Where X a , X b and X c Each is independently selected from monosaccharides.

[0028] In another embodiment related to the first aspect of the present invention, there is also provided a method for preventing, ameliorating or treating URS and GI disorders and / or diseases in a subject in need of such prevention, improvement or treatment, the method comprising:

[0029] i) determining whether the subject suffers from URS and GI conditions and / or diseases, and

[0030] ii) administering to the subject a therapeutically effective amount of a composition comprising an oligosaccharide compound, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compound present in the composition:

[0031] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0032] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0033] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0034] Where X a , X b and X c Each is independently selected from monosaccharides.

[0035] In another embodiment of the present invention, there is provided a use of a composition for preparing a medicament for preventing, ameliorating or treating URS and GI disorders and / or diseases, wherein the composition comprises an oligosaccharide compound, based on the total weight of the oligosaccharide compound present in the composition, the oligosaccharide compound comprises:

[0036] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0037] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0038] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0039] Where X a , X b and X c Each is independently selected from monosaccharides.

[0040] In another embodiment of the present invention, there is provided a composition comprising an oligosaccharide compound for improving athlete performance, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compound present in the composition:

[0041] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0042] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0043] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0044] Where X a , X b and X c Each is independently selected from monosaccharides.

[0045] In another embodiment, a composition for regulating URS and GI environment is also provided, wherein the composition comprises an oligosaccharide compound, based on the total weight of the oligosaccharide compound present in the composition, the oligosaccharide compound comprises:

[0046] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0047] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0048] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0049] Where X a , X b and X c Each is independently selected from monosaccharides.

[0050] As used herein, the terms "treatment", "treating", "treat" and the like refer to obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in terms of completely or partially preventing a disease or its symptoms, and / or therapeutic, in terms of partially or completely curing a disease and / or side effects caused by the disease. As used herein, "treatment" includes any treatment of a disease in mammals, particularly humans, including: (a) preventing the onset of the disease in a subject susceptible to the disease but not yet diagnosed as having the disease; (b) inhibiting the disease, i.e., arresting or slowing its development; and (c) alleviating the disease and / or disease symptoms, i.e., causing regression of the disease.

[0051] The term "subject" as used herein includes any human or non-human animal. The term "non-human animal" includes all mammals, such as non-human primates, sheep, dogs, cats, cows, horses.

[0052] "Therapeutically effective amount" refers to the amount of a compound that, when administered to a subject for treating a disease, is sufficient to achieve treatment for the disease."Therapeutically effective amount" will vary depending on the compound used, the disease and its severity, the age, weight, etc. of the subject to be treated.

[0053] The term "GI disorders and / or diseases" refers to a range of disorders and / or diseases associated with the gastrointestinal tract with recognizable symptoms, including, but not limited to, the following upper GI disorders and / or diseases: dyspepsia, food impaction, dysphagia, heartburn, acid regurgitation, reflux esophagitis, gastroesophageal reflux disease, gastritis, small intestinal bacterial overgrowth, and dyspepsia; and the following lower (or more common) GI disorders and / or diseases: constipation, diarrhea, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), hemorrhoids, anal fissures, perianal abscesses, anal fistulas, diverticular disease or diverticulitis, colitis, infectious colitis, ulcerative colitis, Crohn's disease, ischemic colitis, radiation colitis, pseudomembranous colitis, Clostridium difficile colitis, proctocolitis, amebama, anal spasm, peptic ulcer, colic, biliary colic, gastroenteritis, acrodermatitis enteropathic, intestinal obstruction, intussusception, polyps, Hirschsprung's disease, and diabetes.

[0054] Preferably, the GI disorder and / or disease is an upper GI disorder and / or disease.

[0055] The term "upper respiratory system disorders and / or diseases" or "URS system disorders and / or diseases" refers to a range of disorders and / or diseases associated with the upper respiratory tract with recognizable symptoms, including but not limited to the following respiratory disorders and / or diseases: bacterial infections, viral infections, colds, influenza (flu), respiratory allergies and asthma.

[0056] Preferably, the URS disorder and / or disease is an upper respiratory tract infection (URTI).

[0057] URS or GI disorders and / or diseases may be present in athletes and / or individuals who are (or will be) exercising or training. The composition may be administered before, during, or after exercising or performing physical activity. Alternatively, the composition may be administered in a continuous dosage regimen, such as once or twice a day. The therapeutically effective amount may be a standard dose or a dose determined based on individual factors, such as the age, body mass index (BMI), or total body weight of the individual.

[0058] The terms "modulation" or "modulating" refer to the effect of beneficially adjusting the environment of the GI system or URS to make it more robust and resilient, thereby reducing medical conditions or the severity of these conditions.

[0059] The term “performance” refers to an individual’s ability to perform certain physical tasks associated with sports and exercise, including but not limited to reducing fatigue and improving endurance.

[0060] The term "athlete" refers to an individual who actively engages in sports or exercise, whether professional or amateur.

[0061] The inventors have advantageously found that supplementation with the composition of the present invention reduces the duration of URS, the incidence and severity of GIS, particularly upper GIS, and increases the rate of sIgA secretion in athletes. The composition acts systemically, and studies have shown that the composition of the present invention can modulate immune function and reduce disease, which improves the health of athletes and their availability for training and competition.

[0062] These types of oligosaccharides have a specific connection between the galactose unit (denoted as Gal) and the terminal monosaccharide and are considered to be particularly beneficial for maintaining a healthy human intestinal microbiota and, therefore, for the health of the subject. Known oligosaccharide compositions either do not contain each of these types of oligosaccharides or contain these oligosaccharides in smaller amounts. Therefore, when used as a prebiotic, the composition of the first aspect provides an advantage to the consumer in terms of establishing and maintaining the human intestinal microbiota. Advantageously and unexpectedly, it was found that the composition of the present invention is not only more actively consumed by the microbiota present in the intestine compared to other oligosaccharides, but also in human trials, the composition also showed significantly lower URS and GI symptoms.

[0063] The weight percentage of the specific oligosaccharide compound discussed herein is based on the weight of all oligosaccharide compounds present in the whole composition. Therefore, when determining a specific weight percentage, only the part of the composition provided by the oligosaccharide, component (a), (b), (c) or any other oligosaccharide present is considered. For the determination of oligosaccharide content, in addition to lactose, disaccharides are also included. The composition of this first aspect may contain other non-oligosaccharide components, including monosaccharides and lactose. When determining the specific weight percentage of the oligosaccharide compound discussed herein, these components are not considered. The specific amount of oligosaccharide compound in the composition can be referred to as the weight percentage of the oligosaccharide part in the composition of this first aspect.

[0064] The oligosaccharide compounds (a), (b) and (c) respectively contain X a , X b and X c , each of which is independently selected from carbohydrates. a , X b and X c The groups can be considered as sugar units. Thus, the group X a , X b and X c It can be considered as the terminal sugar of an oligosaccharide. a , X b and X c Any suitable monosaccharide unit capable of forming an oligosaccharide with the galactose units of compounds (a), (b) and (c) can provide X a , X b and Xc Preferably, X a , X b and X c Each independently selected from the following monosaccharides. Suitable monosaccharide units are selected from glucose (Glc), fucose (Fuc), arabinose (Ara), wood sugar (Xyl), rhamnose (Rha), mannose (Man), galactose (Gal), ribose (Rib), lyxose (Lyx), allose (All), altrose (Alt), gulose (Gul), idose (Ido), talose (Tal), psicose (Psi), fructose (Fru), sorbose (Sor), tagatose (Tag), galactosamine (GalN), glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) or mixtures thereof. Therefore, each of compounds (a), (b) and (c) can comprise a mixture of oligosaccharide compounds with different X groups (e.g., X groups are Glc or Fuc).

[0065] In some embodiments, group X a , X b and X c Independently selected from the monosaccharides listed above, preferably independently selected from Glc, Fuc, Ara, Xyl, Rha and Man or a mixture thereof.

[0066] The sugar units of the oligosaccharides in the composition of the first aspect may have the D or L enantiomeric form. Preferably, the Gal sugar units in components (a), (b) and (c) all have the D enantiomeric form. Therefore, components (a), (b) and (c) may be as follows:

[0067] (a)D-Gal-(β1-3)-D-Gal-(β1-4)-X a ;

[0068] (b)D-Gal-(β1-3)-D-Gal-(β1-3)-X b ;and

[0069] (c)D-Gal-(β1-3)-D-Gal-(β1-2)-X c .

[0070] Preferably, Gal and X a , X b and X c The sugar units are all in the D enantiomeric form. Thus, components (a), (b) and (c) may be as follows:

[0071] (a)D-Gal-(β1-3)-D-Gal-(β1-4)-DX a ;

[0072] (b)D-Gal-(β1-3)-D-Gal-(β1-3)-DX b ;and

[0073] (c)D-Gal-(β1-3)-D-Gal-(β1-2)-DX c .

[0074] In some embodiments, X a , X b and X c Each of them is Glc. Therefore, compound (a), (b) and (c) can be galacto-oligosaccharide compound (GOS), and the composition of this first aspect can be called galacto-oligosaccharide composition. These galacto-oligosaccharides can be formed by converting lactose into described oligosaccharides with suitable β-galactosidase. In these embodiments, components (a), (b) and (c) are preferably as follows:

[0075] (a)Gal-(β1-3)-Gal-(β1-4)-Glc;

[0076] (b)Gal-(β1-3)-Gal-(β1-3)-Glc; and

[0077] (c)Gal-(β1-3)-Gal-(β1-2)-Glc.

[0078] Preferably, both Gal and Glc sugar units are in the D enantiomeric form. Thus, components (a), (b) and (c) may be as follows:

[0079] (a) D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glc;

[0080] (b) D-Gal-(β1-3)-D-Gal-(β1-3)-D-Glc; and

[0081] (c) D-Gal-(β1-3)-D-Gal-(β1-2)-D-Glc.

[0082] In some embodiments, X a , X b and X cEach of (a), (b) and (c) is a mixture of Glc and one or more of the above-mentioned other sugar units (e.g., Fuc, Ara, Xyl, Rha and Man). Therefore, each of (a), (b) and (c) may contain a mixture of oligosaccharide compounds having Glc or one of the above-mentioned other sugar units as X groups. In these embodiments, components (a), (b) and (c) may be formed by converting a mixture of lactose and a suitable additional sugar (e.g., a monosaccharide selected from fucose, arabinose, xylose, rhamnose and mannose) into oligosaccharides. Preferably, X a , X b and X c Each of is a mixture of Glc and Fuc.

[0083] In addition to the components (a), (b) and (c) discussed above, other oligosaccharides in the composition of the first aspect comprising oligosaccharide compounds may also comprise the sugar unit "X" groups mentioned above.

[0084] The composition of the first aspect comprises (a) at least 8 wt% of Gal-(β1-3)-Gal-(β1-4)-X a Preferably, the composition comprises at least 9 wt% of component (a) or at least 10 wt% of component (a).

[0085] Preferably, the composition comprises at most 35 wt% component (a), at most 30 wt% component (a) or at most 25 wt% component (a).

[0086] Preferably, the composition comprises 8 wt % to 35 wt % of component (a), 8 wt % to 25 wt % of component (a), or 10 wt % to 20 wt % of component (a).

[0087] The composition of the first aspect comprises (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b Preferably, the composition comprises at least 4 wt% of component (b) or at least 5 wt% of component (b).

[0088] Preferably, the composition comprises at most 25 wt% component (b), at most 20 wt% component (b) or at most 15 wt% component (b).

[0089] Preferably, the composition comprises 3 wt% to 25 wt% of component (b), 4 wt% to 20 wt% of component (b), or 4 wt% to 10 wt% of component (b).

[0090] The composition of the first aspect comprises (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-Xc Preferably, the composition comprises at least 6 wt% of component (c).

[0091] Preferably, the composition comprises at most 25 wt% component (c), at most 20 wt% component (c) or at most 15 wt% component (c).

[0092] Preferably, the composition comprises 5 wt % to 25 wt % of component (c), 5 wt % to 20 wt % of component (c) or 6 wt % to 10 wt % of component (c).

[0093] The above amounts are based on the total weight of the oligosaccharide compounds present in the composition.

[0094] Preferably, in the composition of the first aspect, based on the total weight of the oligosaccharide compounds present in the composition:

[0095] Component (a) is present in an amount of up to 35 wt %;

[0096] Component (b) is present in an amount of up to 25 wt%; and

[0097] Component (c) is present in an amount of up to 25 wt%.

[0098] Preferably, in the composition of the first aspect, based on the total weight of the oligosaccharide compounds present in the composition:

[0099] Component (a) is present in an amount of 8 wt % to 25 wt %;

[0100] Component (b) is present in an amount of 3 wt% to 25 wt%; and

[0101] Component (c) is present in an amount of 5 wt% to 20 wt%.

[0102] In some embodiments, the composition of the first aspect comprises, based on the total weight of the oligosaccharide compounds present in the composition:

[0103] (a) 8 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-4)-Glc;

[0104] (b) 3 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-3)-Glc; and

[0105] (c) 5 to 20 wt% of Gal-(β1-3)-Gal-(β1-2)-Glc.

[0106] Preferably, the weight percentage ratio of compound (a) to compound (b) is 1:1 to 3:1, preferably 1.5:1 to 2.5:1.

[0107] Preferably, the weight percentage ratio of compound (a) to compound (c) is 1:1 to 3:1, preferably 1.5:1 to 2.5:1.

[0108] Preferably, the weight percentage ratio of compound (b) to compound (c) is 2:1 to 1:2, preferably 1.5:1 to 1:1.5.

[0109] Preferably, in the composition of the first aspect, the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compounds present in the composition:

[0110] (d) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-X d ;

[0111] Where X d Selected from sugars.

[0112] X d Can be selected from the above X a , X b and X c of the same monosaccharide units.

[0113] In some embodiments, X d It is Glc.

[0114] Preferably, the Gal sugar unit of component (d) has the D enantiomeric form. Preferably, Gal and X d The sugar units are all in the D enantiomeric form. Therefore, component (d) can be represented as follows: D-Gal-(β1-3)-D-Gal-(β1-3)-D-Gal-(β1-4)-DX d .

[0115] In some embodiments, X d It is a mixture of Glc and one or more other monosaccharide units (eg Fuc, Ara, Xyl, Rha and Man, preferably Fuc).

[0116] Preferably, the composition comprises at least 4 wt% of component (d) or at least 5 wt% of component (d).

[0117] Preferably, the composition comprises at most 25 wt% of component (d), at most 20 wt% of component (d) or at most 15 wt% of component (d).

[0118] Preferably, the composition comprises 3 wt % to 25 wt % of component (d), 4 wt % to 20 wt % of component (d), or 4 wt % to 10 wt % of component (d).

[0119] Preferably, in the composition of the first aspect, based on the total weight of the oligosaccharide compounds present in the composition:

[0120] Component (a) is present in an amount of 8 wt % to 25 wt %;

[0121] Component (b) is present in an amount of 3 wt % to 25 wt %;

[0122] Component (c) is present in an amount of 5 wt% to 20 wt%; and

[0123] Component (d) is present in an amount of 3 wt% to 25 wt%.

[0124] In some embodiments, the composition of the first aspect comprises, based on the total weight of the oligosaccharide compounds present in the composition:

[0125] (a) 8 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-4)-Glc;

[0126] (b) 3 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-3)-Glc;

[0127] (c) 5 wt% to 20 wt% of Gal-(β1-3)-Gal-(β1-2)-Glc; and

[0128] (d) 3 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-Glc.

[0129] Preferably, in the composition of the first aspect, the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compounds present in the composition:

[0130] (e) at least 5 wt% of Gal-(β1-4)-Gal-(β1-4)-X e ;

[0131] Where X e Selected from monosaccharides.

[0132] X e Can be selected from the above X a , X b and X c of the same monosaccharide units.

[0133] Preferably, the Gal sugar unit of component (e) has the D enantiomeric form. Preferably, Gal and X e The sugar units are all in the D enantiomeric form. Therefore, component (e) can be represented as follows: D-Gal-(β1-4)-D-Gal-(β1-4)-DXe .

[0134] In some embodiments, X e It is Glc.

[0135] In some embodiments, X e It is a mixture of Glc and one or more other monosaccharide units (eg Fuc, Ara, Xyl, Rha and Man, preferably Fuc).

[0136] Preferably, the composition comprises at least 6 wt% of component (e) or at least 7 wt% of component (e).

[0137] Preferably, the composition comprises at most 25 wt% of component (e), at most 20 wt% of component (e) or at most 15 wt% of component (e).

[0138] Preferably, the composition comprises 5 wt % to 25 wt % of component (e), 5 wt % to 20 wt % of component (e), or 6 wt % to 10 wt % of component (e).

[0139] Preferably, in the composition of the first aspect, based on the total weight of the oligosaccharide compounds present in the composition:

[0140] Component (a) is present in an amount of 8 wt % to 25 wt %;

[0141] Component (b) is present in an amount of 3 wt % to 25 wt %;

[0142] Component (c) is present in an amount of 5 wt % to 20 wt %;

[0143] Component (d) is present in an amount of 3 wt% to 25 wt%; and

[0144] Component (e) is present in an amount of 5 wt% to 25 wt%.

[0145] In some embodiments, the composition of the first aspect comprises, based on the total weight of the oligosaccharide compounds present in the composition:

[0146] (a) 8 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-4)-Glc;

[0147] (b) 3 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-3)-Glc;

[0148] (c) 5 wt% to 20 wt% of Gal-(β1-3)-Gal-(β1-2)-Glc, and

[0149] (d) 3 wt% to 25 wt% of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-Glc, and

[0150] (e) 5 to 25 wt% of Gal-(β1-4)-Gal-(β1-4)-Glc.

[0151] In the composition of the first aspect, X a , X b , X c , X d and X e Each is independently selected from the above monosaccharides.

[0152] In the composition of the first aspect, X a , X b , X c , X d and X e Each may be independently selected from Glc, Fuc, Ara, Xyl, Rha and Man or a mixture thereof.

[0153] In some embodiments, X a , X b , X c , X d and X e Each is Glc.

[0154] In some embodiments, X a , X b , X c , X d and X e Each contains Fuc. Preferably, X a , X b , X c , X d and X e Each is a mixture of Glc and Fuc.

[0155] The oligosaccharide compound of the composition of the first aspect preferably has a relatively high proportion of β1-3Gal-Gal bonds, mainly due to the presence of components (a), (b), (c) and optionally (d) in the proportions discussed herein. The inventors have found that these relatively high proportions of β1-3Gal-Gal bonds may be particularly advantageous for using the compositions discussed herein.

[0156] In the oligosaccharide compounds, preferably 35%-55% of the Gal-Gal bonds are 1-3 bonds, preferably 40%-55%, preferably 40%-50%.

[0157] In the oligosaccharide compound, preferably 40%-60% of the Gal-Gal bonds are 1-4 bonds, preferably 45%-55%, preferably 45%-52%.

[0158] In the oligosaccharide compounds, preferably 8%-20% of the Gal-X bonds are 1-3 bonds, preferably 10%-18%, preferably 12%-16%, preferably wherein X is Glc.

[0159] In the oligosaccharide compounds, preferably 15%-25% of the Gal-X bonds are 1-4 bonds, preferably 16%-24%, preferably 17%-22%, preferably wherein X is Glc.

[0160] In the oligosaccharide compounds, preferably 30%-45% of the Gal-X bonds are 1-2 bonds, preferably 32%-43%, preferably 34%-41%, preferably wherein X is Glc.

[0161] In the oligosaccharide compounds, preferably 20%-36% of the Gal-X bonds are 1-6 bonds, preferably 22%-34%, preferably 25%-32%, preferably wherein X is Glc.

[0162] Preferably, the above-mentioned Gal-X bond, such as Gal-Glu bond, is a β bond, ie a β-glycosidic bond.

[0163] Preferably, the composition comprises at least 25 wt% trisaccharide, based on the total weight of the composition.Preferably, the composition comprises at least 28 wt% trisaccharide or at least 30 wt% trisaccharide.

[0164] Preferably, the composition comprises at most 70 wt% trisaccharide, preferably at most 60 wt% trisaccharide or at most 50 wt% trisaccharide.

[0165] Preferably, the composition comprises 25 to 70 wt% trisaccharide, preferably 30 to 60 wt% trisaccharide or 30 to 50 wt% trisaccharide.

[0166] Preferably, the composition comprises at least 10 wt% tetrasaccharide based on the total weight of the composition. Preferably, the composition comprises at least 12 wt% tetrasaccharide or at least 15 wt% tetrasaccharide. Preferably, the composition comprises at least 20 wt% tetrasaccharide or at least 25 wt% tetrasaccharide.

[0167] Preferably, the composition comprises at most 40 wt% tetrasaccharide, preferably at most 25 wt% tetrasaccharide or at most 20 wt% tetrasaccharide.

[0168] Preferably, the composition comprises 10 wt% to 40 wt% tetrasaccharide, preferably 10 wt% to 30 wt% tetrasaccharide or 12 wt% to 25 wt% tetrasaccharide.

[0169] Preferably, the composition comprises 30 wt% to 50 wt% of trisaccharides and 10 wt% to 25 wt% of tetrasaccharides, based on the total weight of the composition.

[0170] As mentioned above, the determination of disaccharides present in the composition does not include lactose. Therefore in some embodiments, the composition preferably comprises at most 40wt% disaccharides, preferably at most 30wt% disaccharides or at most 20wt% disaccharides. Preferably, the content of lactose in the composition of the first aspect is minimized. Preferably, the composition is substantially free of lactose. Preferably, the composition is free of lactose.

[0171] Preferably, the composition comprises 10 to 40 wt% disaccharide, preferably 10 to 30 wt% disaccharide or 10 to 20 wt% disaccharide.

[0172] Preferably, the composition comprises 10 wt% to 40 wt% of disaccharides, 30 wt% to 60 wt% of trisaccharides and 10 wt% to 25 wt% of tetrasaccharides, based on the total weight of the composition.

[0173] The following description relates to the content of disaccharides in the oligosaccharide compound.Preferably, the oligosaccharide compound comprises at most 40 wt% disaccharides, preferably at most 30 wt% disaccharides or at most 20 wt% disaccharides.

[0174] Preferably, the oligosaccharide compound comprises 0 wt% to 40 wt% of disaccharides, preferably 10 wt% to 30 wt% of disaccharides or 10 wt% to 20 wt% of disaccharides.

[0175] Preferably, the oligosaccharide compound comprises at least 25 wt% trisaccharides, based on the total weight of oligosaccharide compounds present in the composition.Preferably, the oligosaccharide compound comprises at least 30 wt% trisaccharides or at least 33 wt% trisaccharides.

[0176] Preferably, the oligosaccharide compound comprises at most 75 wt% trisaccharides, preferably at most 65 wt% trisaccharides or at most 55 wt% trisaccharides.

[0177] Preferably, the oligosaccharide compound comprises 25 wt% to 75 wt% of trisaccharides, preferably 30 wt% to 65 wt% of trisaccharides or 34 wt% to 55 wt% of trisaccharides.

[0178] Preferably, the oligosaccharide compound comprises at least 10 wt% tetrasaccharide, based on the total weight of oligosaccharide compounds present in the composition.Preferably, the oligosaccharide compound comprises at least 12 wt% tetrasaccharide or at least 15 wt% tetrasaccharide.

[0179] Preferably, the oligosaccharide compound comprises at most 45 wt% tetrasaccharide, preferably at most 35 wt% tetrasaccharide or at most 30 wt% tetrasaccharide.

[0180] Preferably, the oligosaccharide compound comprises 10 wt% to 45 wt% of tetrasaccharide, preferably 10 wt% to 35 wt% of tetrasaccharide or 15 wt% to 30 wt% of tetrasaccharide.

[0181] Preferably, the oligosaccharide compound comprises 0 wt% to 40 wt% of disaccharides and 30 wt% to 75 wt% of trisaccharides, based on the total weight of the oligosaccharide compounds present in the composition.

[0182] Preferably, the oligosaccharide compound comprises 0 wt% to 40 wt% of disaccharides, 30 wt% to 75 wt% of trisaccharides and 10 wt% to 45 wt% of tetrasaccharides, based on the total weight of the oligosaccharide compounds present in the composition.

[0183] The composition of the first aspect preferably comprises at least 50 wt% of oligosaccharide compounds, including components (a), (b), (c) and optionally (d) and (e). Based on the total weight of the composition, preferably the composition comprises at least 55 wt% of oligosaccharide compounds, preferably at least 60 wt%.

[0184] Preferably, the composition comprises at most 100 wt% of the oligosaccharide compound, preferably at most 95 wt%, at most 90 wt% or at most 85 wt% of the oligosaccharide compound.

[0185] Preferably, the composition comprises 50 wt% to 100 wt% of the oligosaccharide compound, preferably 55 wt% to 95 wt%, or 60 wt% to 85 wt% of the oligosaccharide compound.

[0186] In some embodiments, the composition is in the form of a syrup. The syrup preferably comprises at least 50wt% of an oligosaccharide compound, at least 55wt% or at least 60wt% of an oligosaccharide compound. Preferably, the syrup comprises 50wt% to 75wt% of an oligosaccharide compound, preferably 55wt% to 70wt% or 60wt% to 70wt% of an oligosaccharide compound.

[0187] The syrup may contain a large amount of monosaccharides, such as glucose and / or galactose. The syrup may contain 15wt% to 30wt% monosaccharides, preferably 20wt% to 28wt% monosaccharides or 21wt% to 25wt% monosaccharides, such as glucose and / or galactose.

[0188] The syrup preferably comprises 20 to 30 wt% water, preferably 22 to 28 wt% water.

[0189] The syrup may further contain lactose, for example 4 to 14 wt % lactose.

[0190] In some embodiments, the composition is in the form of a powder. The powder preferably comprises at least 60wt% of an oligosaccharide compound, preferably at least 70wt% or at least 75wt% of an oligosaccharide compound. Preferably, the powder comprises 60wt% to 100wt% of an oligosaccharide compound, preferably 70wt% to 95wt% or 75wt% to 90wt% of an oligosaccharide compound.

[0191] The powder preferably comprises a reduced amount of monosaccharides (e.g. glucose and / or galactose) compared to the above syrup. The powder may comprise 1 wt% to 10 wt% monosaccharides, preferably 2 wt% to 8 wt% monosaccharides, or 3 wt% to 7 wt% monosaccharides, preferably about 5 wt%, such as glucose and / or galactose.

[0192] The powder preferably comprises from 1 to 10 wt% water, preferably from 3 to 6 wt% water.

[0193] The composition of the first aspect may be purified to remove monosaccharides and optionally disaccharides from the composition.

[0194] The composition of the first aspect can be fractionated to separate the oligosaccharide components in the composition according to molecular weight, for example, to remove disaccharides from the composition or to separate trisaccharides from other oligosaccharide components. This can be done by any suitable method known in the art, such as high performance liquid chromatography. The composition produced by this fractionation can be referred to as an oligosaccharide fraction or a GOS fraction.

[0195] In these embodiments, the composition (or oligosaccharide fraction) preferably comprises at least 70 wt% of trisaccharides, tetrasaccharides and higher oligosaccharides, preferably at least 80 wt% or at least 90 wt%. The degree of polymerization of these higher oligosaccharides is 5 and above. Preferably, the composition comprises at least 95 wt% of trisaccharides, tetrasaccharides and higher oligosaccharides. Preferably, the composition consists of or essentially consists of trisaccharides, tetrasaccharides and higher oligosaccharides.

[0196] In these embodiments, the composition preferably comprises 40 to 70 wt% trisaccharide, preferably 45 to 70 wt% trisaccharide or 50 to 70 wt% trisaccharide.

[0197] Preferably, the oligosaccharide compound comprises 15 wt% to 50 wt% of tetrasaccharide, preferably 15 wt% to 40 wt% of tetrasaccharide or 20 wt% to 40 wt% of tetrasaccharide.

[0198] Preferably, the oligosaccharide compound contains 5 wt% to 25 wt% of higher oligosaccharides, preferably 5 wt% to 20 wt% of higher oligosaccharides or 10 wt% to 20 wt% of higher oligosaccharides.

[0199] Based on the total weight of the composition, preferably the composition comprises 40wt% to 70wt% trisaccharides, 15wt% to 40wt% tetrasaccharides and 5wt% to 25wt% higher oligosaccharides.Based on the total weight of the composition, preferably the composition comprises 50wt% to 70wt% trisaccharides, 20wt% to 40wt% tetrasaccharides and 10wt% to 20wt% higher oligosaccharides.

[0200] In these embodiments, the composition (or oligosaccharide fraction) preferably comprises the following amounts of components (a), (b) and (c) as described above, based on the total weight of oligosaccharide compounds present in the composition:

[0201] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0202] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0203] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0204] Where X a , X b and X c Each is independently selected from monosaccharides.

[0205] Preferably, the composition (or oligosaccharide fraction) comprises the following amounts of components (a), (b) and (c) as described above:

[0206] (a) at least 10 wt% of Gal-(β1-3)-Gal-(β1-4)-X a ;

[0207] (b) at least 5 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0208] (c) at least 7 wt% Gal-(β1-3)-Gal-(β1-2)-X c .

[0209] The composition may comprise component (d) and / or component (e) as described above.

[0210] The composition preferably comprises the components in the above-mentioned proportions.

[0211] In some embodiments, the composition of the first aspect is a fractionated product of trisaccharides and tetrasaccharides (which may be referred to as a DP3 / DP4 fraction).

[0212] In these embodiments, the composition (or DP3 / DP4 fraction) preferably comprises at least 70 wt% trisaccharides and tetrasaccharides, preferably at least 80 wt% or at least 90 wt%. Preferably, the composition consists of or consists essentially of trisaccharides and tetrasaccharides.

[0213] In these embodiments, the composition preferably comprises 50 wt% to 80 wt% trisaccharide, preferably 55 wt% to 75 wt% trisaccharide or 60 wt% to 75 wt% trisaccharide.

[0214] Preferably, the oligosaccharide compound comprises 20 wt% to 50 wt% of tetrasaccharide, preferably 25 wt% to 45 wt% of tetrasaccharide or 25 wt% to 40 wt% of tetrasaccharide.

[0215] Based on the total weight of the composition, preferably the composition comprises 50wt% to 80wt% trisaccharides and 20wt% to 50wt% tetrasaccharides.Based on the total weight of the composition, preferably the composition comprises 60wt% to 75wt% trisaccharides and 25wt% to 40wt% tetrasaccharides.

[0216] In these embodiments, the composition (or oligosaccharide fraction) preferably comprises the following amounts of components (a), (b) and (c) as described above, based on the total weight of oligosaccharide compounds present in the composition:

[0217] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0218] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0219] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0220] Where X a , X b and X c Each is independently selected from monosaccharides.

[0221] Preferably, the composition (or oligosaccharide fraction) comprises the following amounts of components (a), (b) and (c) as described above:

[0222] (a) at least 10 wt% of Gal-(β1-3)-Gal-(β1-4)-X a ;

[0223] (b) at least 5 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0224] (c) at least 7 wt% Gal-(β1-3)-Gal-(β1-2)-X c .

[0225] The composition may comprise component (d) and / or component (e) as described above.

[0226] The composition preferably comprises the components in the above-mentioned proportions.

[0227] In some embodiments, the composition of the first aspect is a trisaccharide fractionation product (which may be referred to as a DP3 fraction). This trisaccharide fractionation product can be obtained by the above-mentioned known fractionation method. This composition preferably comprises at least 70wt% trisaccharides, preferably at least 80wt% or at least 90wt%. Preferably, the composition comprises at least 95wt% trisaccharides.

[0228] In these embodiments, the composition (or oligosaccharide fraction) preferably comprises the following amounts of components (a), (b) and (c) as described above, based on the total weight of oligosaccharide compounds present in the composition:

[0229] (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ;

[0230] (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0231] (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ;

[0232] Where X a , X b and X c Each is independently selected from sugars.

[0233] Preferably, the composition (or oligosaccharide fraction) comprises the following amounts of components (a), (b) and (c) as described above:

[0234] (a) at least 10 wt% of Gal-(β1-3)-Gal-(β1-4)-X a ;

[0235] (b) at least 5 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and

[0236] (c) at least 7 wt% Gal-(β1-3)-Gal-(β1-2)-X c .

[0237] The composition may comprise component (e) as described above.

[0238] The composition preferably comprises the components in the above-mentioned proportions.

[0239] The composition of the first aspect can be used as and incorporated into a food supplement product for ingestion by a consumer. Such a product can be selected from dairy products (e.g., liquid milk, milk powder (e.g., whole milk powder, skimmed milk powder, fat-filled milk powder, whey powder, infant formula, ice cream, yogurt, cheese, fermented dairy products), beverages, sports drinks, baby food, cereals, bread, biscuits, candy, cakes, food supplements, dietary supplements, animal feed, poultry feed or any other food or beverage.

[0240] The composition of the first aspect may be incorporated into a synbiotic composition. Such a synbiotic composition is preferably a mixture comprising live microorganisms and substrates selectively utilized by the host microorganisms, which impart health benefits to the host, ie probiotics and prebiotics.

[0241] The composition of the first aspect can be used as a pharmaceutical composition or a nutritional composition comprising at least one carrier, excipient or diluent, and used in the form thereof.

[0242] Suitable further components of pharmaceutical or nutritional compositions and methods of preparing such pharmaceutical or nutritional compositions are known in the art.

[0243] The composition can be administered in a single dose or multiple doses. Suitable frequency of administration can be at least once a day, every other day, once a week, once every two weeks, once every three weeks or four weeks, once a month, once every two months, or once every three to six months. The composition can be administered over a period of at least one week, at least one month, at least three to six months, at least one year, two years, three years, four years, or five years, or during the course of a disease, or throughout the life of the subject.

[0244] It will be apparent to one skilled in the art that administration of the composition will be optimized in clinical trials.

[0245] The composition of the present invention can be formulated into a pharmaceutical composition by combining with an appropriate pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent or a pharmaceutically acceptable excipient, and can be formulated into preparations in solid, semisolid or liquid form, such as tablets, capsules, powders, granules and solutions.

[0246] Pharmaceutically acceptable carriers, excipients or diluents may include, for example, water, saline, glucose, maltodextrin, glycerol, ethanol, salts (e.g., NaCl, MgCl 2 , KCl, MgSO 4 etc.), buffers (e.g., phosphate buffer, citrate buffer, Tris buffer, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), 2-(N-morpholino)ethanesulfonic acid sodium salt (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS) etc.), solubilizing agents; detergents (e.g., nonionic detergents (e.g., Tween-20 etc.)); glycerol, etc.

[0247] Pharmaceutically acceptable carriers, excipients and diluents are non-toxic to the subject at the dosages and concentrations used, and may include, for example, buffers (such as phosphates, citrates and other organic acids); antioxidants (including ascorbic acid, glutathione, cysteine, methionine and citric acid); preservatives (such as ethanol, benzyl alcohol, phenol, m-cresol, p-chloro-m-cresol, methyl or propyl paraben, benzalkonium chloride or a combination thereof); amino acids (such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan); amino acids, methionine, serine, proline and valine and combinations thereof); monosaccharides, disaccharides and other carbohydrates; low molecular weight (less than about 10 residues) polypeptides; proteins (such as gelatin or serum albumin); chelating agents (such as EDTA); sugars (e.g., trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbitol, raffinose, glucosamine, N-methylglucosamine, galactosamine and neuraminic acid); and / or nonionic surfactants (e.g., Tween, Brij Pluronics, Triton-X or polyethylene glycol (PEG)).

[0248] For oral preparations, the composition of the present invention may include suitable additives for making tablets, powders, granules or capsules, for example, adding 30 conventional additives (such as lactose, mannitol, corn starch or potato starch); binders (such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatin); disintegrants (such as corn starch, potato starch or sodium carboxymethyl cellulose); lubricants (such as talc or magnesium stearate); if necessary, diluents, buffers, wetting agents, preservatives and flavoring agents may also be added.

[0249] The pharmaceutical composition may be in liquid form, lyophilized form, or liquid form reconstituted from a lyophilized form, wherein the lyophilized preparation is reconstituted with a sterile solution prior to administration. The standard procedure for reconstitution of a lyophilized composition is to add back a volume of purified water (usually equivalent to the volume removed during the lyophilization process).

[0250] Tonicity agents can be included in the formulation to adjust the tonicity of the formulation. Exemplary tonicity agents include sodium chloride, potassium chloride, glycerol, and the group of any components from amino acids, sugars, and combinations thereof. In some embodiments, the aqueous formulation is isotonic, although hypertonic or hypotonic solutions may also be suitable. The term "isotonic" means that the solution has the same tonicity as other solutions to which it is compared, such as physiological saline or serum.

[0251] The composition may modulate the abundance of a bacterial genus present in the upper gastrointestinal tract. In some embodiments, the composition modulates the abundance of a bacterial genus present in one or both of the small intestine or the large intestine. In some embodiments, the composition modulates the abundance of a bacterial genus predominant in the small intestine selected from the group consisting of Achromobacter, Agrobacterium, Blautia, Burkholderia, Coprococcus, Cryocola, Enterococcus, Eubacterium, Holdemania, Lactococcus, Mycobacterium, Pseudoramibacter, Ralstonia, Sphingomonas, Streptococcus, and Turicibacter.I.

[0252] According to a second aspect of the present invention, there is provided a method for preparing a composition comprising an oligosaccharide compound according to the first aspect, the method comprising the following steps:

[0253] (i) providing a source of sugar compounds;

[0254] (ii) treating the carbohydrate source with one or more β-galactosidases to at least partially convert the carbohydrate source into oligosaccharide compounds.

[0255] Preferably, the steps of the method of the second aspect are performed in the order of step (i) followed by step (ii).

[0256] Preferably, the sugar compound source comprises lactose, lactulose or epilactose. Preferably, the sugar compound source comprises lactose. The sugar source may be lactose, for example lactose syrup from milk. The lactose may be heat treated.

[0257] In some embodiments, no additional sugars (eg, monosaccharides or disaccharides) are added to the sugar source. In these embodiments, the method produces a galacto-oligosaccharide compound.

[0258] In some embodiments, the sugar compound source comprises at least one other sugar. As described above, preferably the at least one other sugar provides an optional terminal monosaccharide unit for the oligosaccharide compound. Preferably, the at least one other sugar is the source of this monosaccharide unit. At least one other sugar as the source of this monosaccharide unit can be a monosaccharide, or can be a higher sugar (such as a disaccharide). The at least one other sugar can be a monosaccharide source selected from glucose (Glc), fucose (Fuc), arabinose (Ara), xylose (Xyl), rhamnose (Rha), mannose (Man), galactose (Gal), ribose (Rib), lyxose (Lyx), allose (All), altrose (Alt), gulose (Gul), idose (Ido), talose (Tal), psicose (Psi), fructose (Fru), sorbose (Sor), tagatose (Tag), galactosamine (GalN), glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) or a mixture thereof. The at least one additional sugar may be one or more of the monosaccharides listed above.

[0259] In these embodiments, the methods produce oligosaccharides having one or more of the above-described monosaccharides as terminal sugar units.

[0260] In some embodiments, the at least one additional sugar may be selected from fucose, arabinose, xylose, rhamnose, mannose or a mixture thereof. The sugar source may comprise lactose and the source of one or more of the monosaccharides. In these embodiments, the method produces an oligosaccharide having a terminal sugar unit selected from Glc, Fuc, Ara, Xyl, Rha and Man or a mixture thereof.

[0261] The step (ii) of the method comprises treating a sugar compound source with at least one galactosidase. The galactosidase may be an alpha-galactosidase or a beta-galactosidase, depending on whether an alpha or beta bond is required between sugar units of the oligosaccharide compound. Preferably, the enzyme exhibits galactosyltransferase (transgalactosidase) activity and forms an alpha- or beta-bond between sugar units in the sugar compound source. This results in the synthesis of an oligosaccharide compound having two or more galactose units derived from lactose. Preferably, step (ii) is performed until the conversion of the sugar source to the oligosaccharide compound is complete.

[0262] Step (ii) may comprise treating the source of carbohydrate compounds with one or more additional enzymes other than β-galactosidase.

[0263] Preferably, the method comprises step (iii) of isolating the β-galactosidase from the composition comprising the oligosaccharide compound.Step (iii) may comprise removing the enzyme by filtration, for example by nanofiltration.

[0264] The composition comprising oligosaccharide compounds produced in step (iii) may be subjected to heat treatment.

[0265] In some embodiments, the composition is evaporated to reduce the water content, thereby providing a final composition comprising an oligosaccharide compound in the form of a syrup, as described above in relation to the first aspect.

[0266] In some embodiments, glucose is removed from the composition produced in step (iii) prior to evaporation. This preferably reduces the glucose content of the composition from 20 wt%-30 wt% to below 10 wt%, preferably to about 5 wt% or less. The water content of the composition is then reduced by evaporation, and the product is dried to provide a powdered final composition comprising an oligosaccharide compound, as described above with respect to the first aspect.

[0267] It should also be understood that those skilled in the relevant art can produce a composition comprising oligosaccharide compounds according to the first aspect, ie, a composition comprising a specified amount of a specific oligosaccharide compound, by combining the oligosaccharides obtained and isolated from different sources in desired amounts.

[0268] The present invention is described below, by way of example only, with reference to the accompanying drawings, in which:

[0269] Figure 1 A graph showing the results of butyrate analysis of three donors after administration of oligosaccharides of the present invention and comparative oligosaccharides;

[0270] Figure 2A graph showing weekly GIS scores reported during the 24-week study (compositions containing oligosaccharide compounds of the present invention (hereinafter also referred to as B-GOS) n=16, placebo n=17), data are expressed as mean ± SD. AUC analysis showed differences between the groups (P<0.05);

[0271] Figure 3 Figure 2 shows salivary IgA concentrations before and after 12 and 24 weeks of the study (B-GOS n=16, placebo n=17), data are expressed as median (IQR). Asterisks (*) indicate significant differences between groups (P<0.05); and

[0272] Figure 4 Graphs showing salivary IgA secretion rates before and after 12 and 24 weeks of the study (B-GOS n=16, placebo n=17), data are expressed as mean±SD. Asterisks (*) indicate significant differences between groups (P<0.05). Example

[0273] Example 1: Oligosaccharide syrup

[0274] The composition comprising oligosaccharide compounds according to the present invention is obtained in the form of syrup by the following steps. Lactose is rehydrated with drinking water to obtain a working solution with a solid content between 35wt% and 65wt%. The lactose solution is heat treated and then cooled to 40°C-65°C. The pH value of the solution is adjusted to 5.5-7.5. β-galactosidase is then added to the solution in a closed container and then reacted with lactose to catalyze the transfer of galactose molecules to produce oligosaccharide compounds. The progress of the reaction is monitored by measuring the generation of glucose. The reaction is carried out for 8 to 26 hours. The reaction is then terminated by high temperature treatment. The reaction mixture is cooled and the enzyme is removed by carbon filtration. The mixture is then dried by evaporation to reduce the water content to about 22wt%-28wt%, thereby providing a product in the form of syrup.

[0275] Example 2: Oligosaccharide powder

[0276] The composition comprising oligosaccharide compounds according to the present invention is obtained in powder form by the modification of the above method. After removing the enzyme, the mixture is further filtered to remove most of the glucose and other monosaccharides, reducing the monosaccharide content from about 23wt% to about 5wt%. The water content of the composition is then reduced by evaporation, and the product is dried to obtain a powder having a water content of about 3wt%-6wt%.

[0277] Example 3: Classification

[0278] A 5×70 cm BioGel P2 column was used to separate different fractions (DP2, DP3, DP4 or DP5) from the sample of Example 2 using water as the eluent. The column was run at a flow rate of 40 mL / h to 100 mL / h at room temperature (21° C.). Depending on the run, 0.5, 0.75, 1.0, 1.5 or 2.0 mL of a 0.5 g / mL solution of Example 2 dissolved in ultrapure water was added. The column was loaded a total of 15 times with this solution to obtain sufficient low-abundance fraction material, i.e., DP5. After a void volume of approximately 725 mL, 5 mL fractions were collected. Thin layer chromatography (TLC) and HPAEC-PAD were used for analysis to ensure that molecules with the same DP were merged.

[0279] Pooled fractions were frozen and lyophilized. Dry matter from all runs was combined and reconstituted. After the second freezing and lyophilization cycle, the dry matter was stored at 4°C for further analysis or experiments.

[0280] Comparative Example 1

[0281] A commercially available composition comprising an oligosaccharide compound is obtained in powder form.

[0282] Oligosaccharide analysis

[0283] The samples of Example 2 of the present invention and Comparative Example 1 were analyzed by the following steps to determine their oligosaccharide contents.

[0284] Materials and methods

[0285] The dried powder samples of Example 2 and Comparative Example 1 were respectively dissolved in water to provide solutions having a concentration of 40 g / l for analysis.

[0286] Gel Permeation Chromatography

[0287] An HPLC apparatus equipped with a Rezex RSO and RI detector and online desalting (for removal of salts and charged species such as proteins) was used for aqueous GPC separation of sample components. Separation was performed at high temperature (80°C). The separation range of the Rezex RSO column is from DP1 (monosaccharides) to approximately DP10. All samples were analyzed undiluted (40 g / L). Prior to analysis, all sample solutions were treated at 100°C for 10 minutes to remove any microbial or enzymatic activity.

[0288] GOS fingerprint

[0289] HPAEC-PAD (high-performance anion exchange chromatography) equipped with a PA-1 column is used to separate monosaccharides and oligosaccharides of different samples. An attempt was made to achieve the separation quality described in van Leeuwen et al., Carbohydrate Research 2016, 425, 48-58. Commercial maltooligosaccharide mixtures and comparative example 1 were also injected to compare the chromatograms with those reported by Van Leeuwen et al. Based on this, many peaks were annotated. The samples for GPC were diluted 100 times with DMSO before injection.

[0290] result

[0291] Gel Permeation Chromatography (GPC-RI)

[0292] Table 1 shows the DP (degree of polymerization) composition results for samples using the Rezex-RSO system, based on RI calibration with glucose (values ​​in samples are expressed as g / L). All material eluting in the >DP5 window was combined.

[0293] Table 1

[0294]

[0295] The concentration information can be used to calculate the relative weight percentages of different DP fractions of oligosaccharides contained in the sample, as shown in Table 1, where DP=2 refers to disaccharides, DP=3 refers to trisaccharides, and so on.

[0296] GOS fingerprint

[0297] To identify individual galacto-oligosaccharides in the samples (GOS fingerprint), a gradient was developed that gave a separation comparable to that reported in van Leeuwen et al., Carbohydrate Research 2016, 425, 48-58. Peak annotations were made in the chromatograms of all GOS samples based on the peak annotations of oligosaccharide compounds by van Leeuwen et al. A retention window of approximately 15 seconds was used for peak annotations.

[0298] Table 2 shows the HPAEC-PAD peak area information for all annotated peaks, as well as the incubation conditions, sample concentrations, dilutions, and injection volumes.

[0299] Table 2

[0300]

[0301]

[0302] Assume that the peak area corresponds approximately to the amount of each oligosaccharide compound present in the composition. If no specific oligosaccharide compound is identified, enter "unknown" and a number in the table. The identified compound is identified by name or number, which corresponds to the number assigned to a specific galacto-oligosaccharide in van Leeuwen et al., Carbohydrate Research 2016, 425, 48-58. A list of these galacto-oligosaccharides and their corresponding numbers is provided below.

[0303] The above oligosaccharide components (a)-(e) correspond to the following numbers in Table 2 above, wherein each X group is Glu:

[0304] (a)D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glu-12

[0305] (b)D-Gal-(β1-3)-D-Gal-(β1-3)-D-Glu-29

[0306] (c)D-Gal-(β1-3)-D-Gal-(β1-2)-D-Glu-30

[0307] (d)D-Gal-(β1-3)-D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glu-31

[0308] (e) D-Gal-(β1-4)-D-Gal-(β1-4)-D-Glu-11.

[0309] Prebiotic Effect: Butyrate Production

[0310] Butyrate is produced by the intestinal microbiota from the conversion of acetate and / or lactate (and other substrates) to butyrate. Since butyrate is a secondary metabolite, it is usually produced in the later stages of incubation. These experiments were designed to evaluate the differences in butyrate production between the compositions of the present invention and the comparative GOS compositions to understand whether they are more effective as prebiotics. The experiments used the following procedure.

[0311] The composition of Example 2 of the present invention, Comparative Example 1 and a control blank sample were dialyzed using a 0.5 kDa membrane to provide a 5 g / L sample, which was then mixed with fecal material obtained from three healthy adult subjects (donors A, B and C). The mixture was shaken under anaerobic conditions, and colonic fermentation products, including butyrate, were monitored over 48 hours (collection points were 6, 24 and 48 hours). During this period, the distribution of oligosaccharides in the mixture was also monitored using the method described in Table 2 above. The results showed that Example 2 was fully fermented by all donors primarily during the 0-24 hour period, and the butyrate production increased at the 6 and 48 hour time points compared to Comparative Example 1 and the control. The analysis results of butyrate are shown in Figure 2. Figure 1 The results of oligosaccharide analysis of each sample at different time points are shown in Table 3. These results indicate that during the experiment, the oligosaccharides in the samples were actively consumed by the microbiota present in the fecal samples.

[0312] In order to evaluate whether the treatment effect on intestinal microbial activity is statistically significant, three two-sided T tests were performed between Example 2 and the control, Comparative Example 1 and the control, and Example 2 and Comparative Example 1 to obtain p values. The Benjamini-Hochberg false discovery rate (FDR) was also used for analysis. When the p value obtained was less than the reference value, the difference between the treatment effects was considered significant. Table 4 below shows the difference in average butyrate production of the comparison samples within 48 hours, and the asterisk indicates whether the difference is considered significant based on the above analysis. These results show that the increase in butyrate production provided by Example 2 during 0-48 hours is statistically significant compared to the control and Comparative Example 1.

[0313]

[0314] Table 4

[0315] D0-48h / output Example 2 - Control Comparative Example 1-Control Example 2-Comparative Example 1 Butyrate (mM) 5.88* 4.75* 1.13*

[0316] In summary, the present invention provides a composition comprising an oligosaccharide compound (e.g., a galacto-oligosaccharide compound), which comprises, based on the total weight of the oligosaccharide compounds present in the composition: (a) at least 8 wt% of Gal-(β1-3)-Gal-(β1-4)-X a (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b and (c) at least 5 wt% Gal-(β1-3)-Gal-(β1-2)-X c ; where X a , X b and X cEach is independently selected from monosaccharides. Compared with known oligosaccharide compositions, these compositions contain relatively high content of oligosaccharide compounds (a), (b) and (c) and relatively high content of β1-3Gal-Gal bonds. These specific features of the composition are believed to be beneficial to the intestinal health of the consumer, for example, because these compositions increase the production of butyrate in the intestinal tract of the consumer compared with known compositions.

[0317] Example 4: Effects of 24-week prebiotic intervention on self-reported upper respiratory tract, gastrointestinal symptoms and immune markers in elite rugby league players

[0318] This study evaluated the effects of 24 weeks of B-GOS supplementation on the severity, duration and incidence of URS and GIS, salivary immunoglobulin A (sIgA), and C-reactive protein (CRP) and TNF-α concentrations in elite rugby league players during the season.

[0319] This study was a randomized, double-blind, placebo-controlled trial conducted over a 24-week period during the regular rugby league season in the Gallagher English Premier League. Forty-one healthy elite rugby league players (mean age 23.5±4.7 years; weight 103.8±13.2 kg; 186.5±7.8 cm) from a single club volunteered to participate in this study. All individuals were non-smokers, had no history of gastrointestinal disease, and did not regularly consume foods rich in probiotics, prebiotics, or vitamins. Participants were paired based on weight and playing position prior to randomization to the intervention (see Table 5 below). All data were collected during the UK autumn and winter months (temperature range -4 to +25°C).

[0320] During the study period, a typical week for participants consisted of four to five days of training (lasting five hours each day), one match, and at least one day of rest. Training included endurance, skills, fitness, tactics, and match practice. Athletes who were not included in the match roster would receive an additional training day. Participants did not follow a personalized diet plan, but rather meals were provided on-site during training. A cooked breakfast was provided before the first training session, and a snack was provided after the morning session. The afternoon session was followed by the main meal. Similar meals were provided before the competition, and recovery drinks were provided after the competition. Similarly, no meal plan was provided upon leaving the training ground. However, individuals were instructed to avoid any foods rich in probiotics, prebiotics, and vitamins. Prior to each data collection visit, individuals were asked to arrive after an overnight fast and to avoid using mouthwash.

[0321] Supplements

[0322] Athletes were randomly assigned to consume 2.9 g of the oligosaccharide composition of the present invention (in powder form) or 2.9 g of a placebo (maltodextrin) per day. The two supplements were identical in taste and color and were blinded to which supplement was taken. Researchers, club staff, and participants were blinded until all statistical analyses were completed. Participants were provided with the supplement pre-mixed in water at the training ground and drank it under the observation of club staff. On rest days, athletes were instructed to mix the sachets into water and consume them at breakfast. Participants returned used and unused sachets to assess compliance with the supplement.

[0323] Daily upper respiratory symptoms

[0324] To determine the presence of URS, participants completed the Jackson Questionnaire (Jackson et al., 1958) daily. The presence of eight symptoms (headache, chills, sneezing, sore throat, fatigue, cough, runny nose, nasal congestion) was rated on a 0-3 scale (0-none, 1-mild, 2-moderate, 3-severe). All symptom scores for each day were summed to obtain a total Jackson symptom score. URS episodes were defined using the Jackson criteria applied by Martineau, Hanfia, and Witt (2015), with an episode defined as any duration ≥3 days, a Jackson score ≥14, the presence of a runny nose or a symptom score <14, and a subjective feeling of having a cold for at least 3 days. If URS symptoms recurred within a week, it was considered the same episode.

[0325] Weekly Gastrointestinal Symptoms

[0326] To assess gastrointestinal symptoms (GIS), participants were asked to complete a weekly GI symptom tool (Gaskell et al., 2019). Participants were trained and informed to rate each symptom using a 10-point visual analog scale, with 1-4 indicating mild GIS (i.e., a feeling of GIS, but not enough to interfere with exercise), 5-9 indicating severe GIS (i.e., GIS enough to interfere with exercise), and 10 indicating extremely severe GIS (i.e., causing a reduction in exercise effort or cessation of exercise). If no specific GIS was reported, this was scored as zero. GIS, including regurgitation, projectile vomiting, and defecation, were scored as 0 or 10, as the presence of these symptoms would lead to cessation of exercise. Participants were asked to rate each symptom with reference to the previous 7 days. All symptom scores were summed to produce a weekly total score and incidence. In addition, the maximum possible weekly score for each individual was 190 (Gaskell et al., 2019).

[0327] Collection and analysis of sIgA

[0328] At weeks 0, 12, and 24, all participants provided saliva samples for the measurement of sIgA. Participants rinsed their mouths with tap water and remained seated for 10 minutes before providing each sample. Unstimulated saliva samples were produced using a passive method, with participants instructed to collect as much saliva as possible in their mouths for 2 minutes and then slowly release saliva into a collection tube by tilting their heads forward for another 2 minutes. Samples were immediately frozen at -20°C and then frozen at -80°C within 48 hours until analysis. For analysis, samples were completely thawed at room temperature and sIgA concentrations were measured by enzyme-linked immunosorbent assay (ELISA) (Salimetrics, Philadelphia, PA). For sIgA, the intra-assay and inter-assay coefficients of variation were 3.2% and 7.3%, respectively, and the minimum detectable level of the assay was 2.5 μg / mL, which all samples exceeded. sIgA flow rate was calculated by multiplying the concentration of sIgA (μg / mL) by the flow rate (mL / min) to give the concentration per unit time (μg / min) as determined according to the manufacturer's instructions.

[0329] Collection and analysis of blood biomarkers of systemic inflammation

[0330] All participants provided blood samples at weeks 0, 12, and 24. Blood samples were drawn from the antecubital vein in two 10-mL blood collection tubes, one containing heparin and the other containing EDTA coagulant (BD ). Plasma was used to determine the concentrations of tumor necrosis factor-α (TNF-α) and CRP at each time point. The samples were centrifuged and immediately frozen at -20°C and then frozen at -80°C within 48 hours until analysis. TNF-α was assessed using a high-sensitivity ELISA, and CRP was assessed using a conventional ELISA protocol (R&D systems). For TNF-α, the intra-assay coefficient of variation and the inter-assay coefficient of variation were 7.5% and 5.8%, and the minimum detectable level of the assay was 0.022 pg / ml, which was exceeded by all samples. For CRP, the intra-assay coefficient of variation and the inter-assay coefficient of variation were 4.4% and 7.8%, and the minimum detectable level of the assay was 0.022 pg / ml, which was exceeded by all samples.

[0331] Statistical analysis

[0332] Statistical analysis was performed using the Statistical Package for Social Sciences (IBM SPSS version 26, Illinois, USA). All data were tested for normal distribution using the Shapiro-Wilk test. The area under the curve (AUC) of daily URS and weekly GIS over 24 weeks was compared using the Mann-Whitney U test to assess differences in symptom severity. The Mann-Whitney U test was used to assess differences in the incidence of URS between groups. An independent t-test was used to assess differences in the duration of URS onset and GIS-free weeks. Analysis of covariance (ANCOVA) was used to assess the secretion rates of CRP, TNF-α, and sIgA, adjusting for baseline values. Data evaluated using parametric tests are expressed as mean standard deviations. Data presented using nonparametric tests are presented as medians (ranges). Statistical significance was set at P<0.05.

[0333] Athlete characteristics

[0334] The final sample consisted of 33 participants (placebo n = 17, B-GOS n = 16) who successfully completed the full 24-week supplementation period. Adherence to supplementation in training and at home was good (B-GOS: 80.4 ± 13.9%, placebo: 78.3 ± 14%, P = 0.73). There were no differences in weight, height, age, average weekly workload, and playing time between the groups (P > 0.05) (see Table 5 for characteristics of the study participants).

[0335] Table 5

[0336] factor B-GOS[SB1] Placebo p-value Random N (%) 16(48％) 17(52％) Age (years), mean (SD) 22.4(3.3) 24.5(5.2) 0.16 Height (cm), mean (SD) 186.9(9.4) 186.6(7.3) 0.91 Body weight (kg), mean (SD) 103.4(14.0) 105.2(13.2) 0.71 BMI, mean (SD) 29.5(2.4) 30.2(3.1) 0.49 Unit-Back 8(50％) 7(41％) 0.61 Unit - Forward 8(50％) 10(59％) 0.61 Weekly workload (AU), mean (SD) 672.4(129.6) 675.9 0.93 Game time, mean (SD) 657.4(279.5) 597.9 0.57

[0337] Upper respiratory tract symptoms (URS)

[0338] The Mann-Whitney U test showed no difference in the incidence of URS between B-GOS (1.0±1.4) and placebo (1.0±1.0) (P=0.64) (see Table 6 for an overview of self-reported URS data). The AUC of daily symptom scores did not differ between the two groups (P=0.77). The duration of individual URS episodes was shorter in the oligosaccharide group (7.4±2.8 days) compared with the placebo group (9.8±4.1 days) (P=0.04).

[0339] Table 6

[0340]

[0341] Gastrointestinal symptoms (GIS)

[0342] There was no significant difference in GIS between the two groups in the first 7 days before day 0 (P = 0.53). Mann-Whitney U test showed that the AUC of the weekly total symptom score was lower in the B-GOS group (50 [10.5-139.5]) compared with the placebo group (149 [69-208]) (P = 0.03) ( Figure 2 ). The AUC weekly upper GIS symptom score was lower in the B-GOS group compared with the placebo group (P<0.001), but no difference was found in the lower GIS (P=0.11) (data not shown). The number of symptom-free weeks for total GIS and upper GIS was lower in the B-GOS group compared with the placebo group (P<0.05), and there was no significant difference in the number of symptom-free weeks for the lower GIS (P=0.15) (see Table 7, showing the number of GIS symptom-free weeks during the 24-week study period).

[0343] Table 7

[0344]

[0345] Systemic inflammation

[0346] There was no difference in CRP and TNF-α between the two groups at any time point (P>0.05).

[0347] sIgA

[0348] No differences in sIgA concentrations were observed between the two study groups at week 0 or week 12, but at week 24, sIgA was higher in the B-GOS group compared with the placebo group (B-GOS, median (IQR) 245 (177.9, 319.2), (placebo median (IQR) 169 (110.6, 229.9); P = .006) ( Figure 3 No differences in sIgA secretion rates were observed at week 0 or week 12, but a difference was observed at week 24 (B-GOS, median (IQR) 142 μg / min (93.9, 155.0) vs (placebo, 86 μg / min (71.5, 105.5)); P = 0.01)( Figure 4 ).

[0349] Conclusions of prebiotic intervention

[0350] The main findings of this study were that daily supplementation with B-GOS reduced the duration of URS and the incidence of GI symptoms in elite rugby league players over a 24-week period. In addition, B-GOS increased sIgA concentrations and secretion rates at 24 weeks compared with the placebo group. These findings suggest that B-GOS can modulate the immune and GI systems and inhibit URS and GI discomfort associated with elite rugby league play.

[0351] Illness can adversely affect an athlete's training effectiveness and readiness for competition (Cunniffe et al., 2009; Tiernan et al., 2020; Keaney et al., 2021). To the best of the inventors' knowledge, this is the first study to evaluate the effects of a prebiotic dietary intervention on URS and GI in an athlete population. A 22% reduction in the duration of URS episodes is clinically meaningful and relevant to athletes and coaches, ensuring that athletes can return to competition more quickly after an episode of URS.

[0352] Previous studies have shown that B-GOS can promote the growth of bifidobacteria in the human gut, resulting in numerous health benefits, such as reduced systemic inflammation and improved immune responses in elderly and overweight individuals (Vulevic et al., 2008; Vulevic et al., 2013). In addition, increases in fecal short-chain fatty acid (SCFA) production, as well as potential enhancement of epithelial integrity and mucosal immunity have been reported (Hernot et al., 2009; Mariadason, Barkla & Gibson, 1997). Indeed, in the current study, B-GOS increased sIgA concentrations and secretion rates at 24 weeks, a time when immunity may be compromised due to increased training load, as observed in the reduction in the placebo group. This suggests that active manipulation of the gut microbiome may support mucosal immunity and sIgA production.

[0353] B-GOS is believed to reduce systemic inflammation by increasing the growth of Bifidobacterium, which may expand the population of dendritic cells in the intestine, alter naive T cell differentiation, and increase T cells expressing forkhead box protein P3 (FoxP3), thereby producing an anti-inflammatory effect (McLoughlin & Mills, 2011).

[0354] It should be noted that the total number of URS episodes in this study was lower than previously reported for winter. Elite rugby league players have been reported to experience four URS episodes per season (Cunniffe et al., 2009), with the highest incidence in the winter. Our report demonstrated a significantly lower incidence, but with a higher incidence early in the season. This could explain why B-GOS had little effect on the incidence and severity of URS. However, it does suggest that B-GOS may improve the duration of URS even at a low incidence. We also found that participants continued to train despite URS. This finding has been found in other similar cohorts and may be due to participants being asked to train while at the training ground (Cunniffe et al., 2009). They may also have feared being left out of upcoming matches. This is a limitation of collecting self-reported data, and future studies should use molecular testing to determine infection. Another limitation is the frequency of sIgA and cytokine measurements. B-GOS may have shortened the duration of URS by enhancing sIgA.

[0355] In conclusion, 24 weeks of prebiotic B-GOS supplementation reduced the duration of URS, the incidence and severity of GIS, and increased the rate of sIgA secretion in elite rugby league players. These findings suggest that B-GOS has the potential to modulate immune function and reduce disease, which could improve the effectiveness of athlete training and competition.

[0356] While several preferred embodiments have been shown and described, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the scope of the invention as defined in the appended claims.

[0357] Throughout the specification, the term "comprising" or "comprises" means including the specified components, but does not exclude the presence of other components. The term "consisting essentially of" or "consists essentially of" means including the specified components, but excluding other components, except for substances present as impurities, substances that are inevitably present due to the method of providing the components, and components added for purposes other than achieving the technical effects of the present invention. Generally, when referring to a composition, a composition consisting essentially of a series of components will contain less than 5wt%, generally less than 3wt%, and more generally less than 1wt% of non-specified components.

[0358] The term "consisting of" or "consists of" means including the specified components but excluding the addition of other components.

[0359] Where appropriate, depending on the context, the terms “comprises” or “comprising” used may also be understood to encompass or include the meaning of “consists essentially of” or “consisting essentially of” and may also be understood to include the meaning of “consisting of” or “consists of.”

[0360] For the avoidance of doubt, when the amount of a component in a composition is described in wt%, this means the weight percentage of the specified component relative to the entire composition. For example, "wherein the oligosaccharide compound comprises at most 35 wt% of disaccharides" means that 35 wt% of the oligosaccharide compound in the composition is provided by disaccharides.

[0361] Where appropriate, the optional features set forth herein may be used alone or in combination with each other, particularly in the combinations set forth in the appended claims. Where appropriate, the optional features of each aspect or exemplary embodiment of the invention set forth herein should also be understood to be applicable to any other aspect or exemplary embodiment of the invention. In other words, the technician reading this specification should consider the optional features of each exemplary embodiment of the invention to be interchangeable and combinable between different exemplary embodiments.

[0362] Please note that all papers and documents related to this application filed concurrently or previously with this specification are open to public inspection together with this specification, and the contents of all these papers and documents are incorporated into this specification by reference.

[0363] All features disclosed in this specification (including any accompanying claims and drawings), and / or all steps of any disclosed method or process, may be combined in any combination, except combinations in which at least some of such features and / or steps are mutually exclusive.

[0364] Unless expressly stated otherwise, each feature disclosed in this specification (including any attached claims and drawings) may be replaced by alternative features for the same, equivalent or similar purpose. Therefore, unless expressly stated otherwise, each feature disclosed is only an example of a series of equivalent or similar features.

[0365] The invention is not limited to the details of the foregoing embodiments. The invention extends to any novel one or any novel combination of the features disclosed in this specification (including any attached claims and drawings), or to any novel one or any novel combination of the steps of any disclosed method or process.

Claims

1. A composition comprising an oligosaccharide compound for preventing, improving or treating upper respiratory system (URS) and gastrointestinal tract (GI) disorders and / or diseases, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compound present in the composition: (a) at least 8 wt% Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-X b ;and (c) at least 5 wt% of Gal-(β1-3)-Gal-(β1-2)-X c ; Where X a , X b and X c Each is independently selected from monosaccharides.

2. A composition for use according to claim 1, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compounds present in the composition: (d) at least 3 wt% of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-X d ; Where X d Selected from monosaccharides.

3. A composition for use according to claim 1 or 2, wherein based on the total weight of the oligosaccharide compounds present in the composition: Component (a) is present in an amount of up to 35 wt %; Component (b) is present in an amount of up to 25 wt%; and Component (c) is present in an amount of up to 25 wt%.

4. A composition for use according to any one of the preceding claims, wherein the weight percentage ratio of compound (a) to compound (b) is from 1:1 to 3:

1.

5. A composition for use according to any one of the preceding claims, wherein the weight percentage ratio of compound (a) to compound (c) is from 1:1 to 3:

1.

6. A composition for use according to any one of the preceding claims, wherein the weight percentage ratio of compound (b) to compound (c) is from 2:1 to 1:

2.

7. A composition for use according to any one of the preceding claims, wherein the oligosaccharide compound comprises, based on the total weight of the oligosaccharide compounds present in the composition: (e) at least 5 wt% of Gal-(β1-4)-Gal-(β1-4)-X e ; Where X e Selected from monosaccharides.

8. A composition for use according to any one of the preceding claims, wherein X a , X b , X c , X d and X e Each is independently selected from glucose, fucose, arabinose, xylose, rhamnose, mannose, galactose, ribose, lyxose, allose, altrose, gulose, idose, talose, psicose, fructose, sorbose, tagatose, galactosamine, glucosamine and N-acetylglucosamine or a mixture thereof.

9. A composition for use according to any one of claims 1 to 7, wherein X a , X b , X c , X d and X e Each is Glc.

10. A composition for use according to any one of claims 1 to 7, wherein X a , X b , X c , X d and X e Each contains Fuc.

11. A composition for use according to any preceding claim, wherein 40% to 55% of the Gal-Gal bonds in the oligosaccharide compound are 1 to 3 bonds.

12. A composition for use according to any preceding claim comprising at least 50 wt% of an oligosaccharide compound, preferably wherein the composition is in the form of a syrup.

13. A composition for use according to any preceding claim comprising at least 75 wt% of an oligosaccharide compound, preferably wherein the composition is in powder form.

14. A composition for use according to any preceding claim, wherein the oligosaccharide compound comprises at least 25 wt% trisaccharides.

15. A composition for use according to any preceding claim, wherein the oligosaccharide compound comprises at least 10 wt% tetrasaccharides.

16. A composition for use according to any preceding claim comprising 80 wt% of trisaccharides, tetrasaccharides and higher oligosaccharides.

17. A composition for use according to any preceding claim comprising 80 wt% trisaccharide.

18. A composition for use according to any one of the preceding claims, wherein the composition is in the form of a dietary supplement.

19. A composition for use according to any one of claims 1 to 17, wherein the composition is in the form of a pharmaceutical or nutritional composition.

20. The composition for use according to claim 19, wherein the pharmaceutical or nutritional composition further comprises at least one carrier, excipient or diluent.

21. A composition for use according to any preceding claim, wherein the upper gastrointestinal tract disorder and / or disease is selected from one or more of the following upper gastrointestinal tract disorders and diseases: dyspepsia, food impaction, dysphagia, heartburn, acid reflux, reflux esophagitis, gastroesophageal reflux disease, gastritis, small intestinal bacterial overgrowth and dyspepsia.

22. A composition for use according to any one of the preceding claims, wherein the upper respiratory system disorder and / or disease is an upper respiratory tract infection (URTI).

23. Composition for use according to any of the preceding claims, wherein the upper respiratory system and gastrointestinal tract disorders and / or diseases are athletes and / or individuals undergoing or about to undergo physical exercise.

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