Composition of galactooligosaccharide and its use

Abstract

Prebiotic oligosaccharide compositions for use in the prevention, amelioration or treatment of upper respiratory system (URS) and upper gastrointestinal (GI) conditions and / or diseases. The compositions comprise oligosaccharide compounds, for example, based on the total weight of the oligosaccharide compounds present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c , wherein X a , X b and X c are each independently selected from monosaccharides, and comprise galactooligosaccharide compositions. These compositions contain relatively high amounts of oligosaccharide compounds (a), (b) and (c) and relatively high amounts of β1-3 Gal-Gal linkages compared to known oligosaccharide compositions. The present invention also relates to compositions comprising oligosaccharide compounds for enhancing the performance of athletes. These specific features of the compositions are thought to provide benefits to the health of the consumer's gut and to reduce, for example, the symptoms of URS and upper GI conditions and / or diseases. Also disclosed are methods of preparing said compositions and the use of said compositions as nutritional supplements or medicaments.

Description

Technical Field

[0001] Field The present invention relates to compositions comprising oligosaccharide compounds for use in the prevention, amelioration or treatment of upper respiratory system (URS) and gastrointestinal (GI) conditions and / or diseases. The present invention also relates to compositions comprising oligosaccharide compounds for enhancing the performance of athletes. The present invention also relates to methods of preparing said compositions and formulations of said compositions as nutritional supplements and / or medicaments. In particular, the present invention relates to oligosaccharide compositions having a relatively high amount of certain beneficial oligosaccharides.

Background Art

[0002] Background The human gut microbiota (or microflora) includes genera of pathogenic, benign and beneficial microorganisms. The predominance of the former can lead to intestinal disorders that can be both acute (e.g., gastroenteritis) and chronic (e.g., inflammatory bowel disease, irritable bowel syndrome and some intestinal cancers). In humans, the colonic microbiota is acquired at birth. Infants breastfed have a predominance of Bifidobacterium, which easily outcompetes bacteria of other genera. This is because the components of human milk stimulate the growth of Bifidobacterium. However, many factors, such as the environment, drug treatments and diet, especially the consumption of highly processed foods, can affect the composition and activity of an individual's intestinal microbiota over their lifetime (see “Role of the gut microbiota in nutrition and health”, Valdes, A.M. et al., BMJ, 2018, 361).

[0003] Certain components of the gut microbiota are involved in the etiology of intestinal diseases. For example, Mycobacterium is associated with Crohn's disease, ulcerative colitis can be caused by sulfate-reducing bacteria, and there may be a bacterial involvement in the development of intestinal cancer. It would be beneficial if the selective growth of indigenous beneficial gut bacteria could be promoted by the intake of prebiotics. Prebiotics are substrates that are selectively utilized by host microbes to confer health benefits. The use of such prebiotics may have a continuing beneficial effect of suppressing the growth of pathogenic microbiota.

[0004] Attempts have been made to favorably influence the balance of the gut microbiota in favor of beneficial microorganisms, such as Bifidobacterium, by adding one or more such microbial strains to a suitable food medium with the intention of conferring health benefits on the host. Such dietary supplements of live microorganisms are known as probiotics. However, it is difficult to guarantee the survival of live bacteria in food after digestion, and as a result, the actual effects of probiotics can be limited. Therefore, prebiotics may be a more promising option for positively influencing the gut microbiota in patients (see Walter J, Maldonado-Goemez MX, Martinez I. “To engraft or not to engraft: an ecological framework for gut microbiome modulation with live microbes”, Curr Opin Biotechnol. 2018, 49, 129-139).

[0005] As noted above, an alternative approach to dietary manipulation of the gut microbiota is the use of prebiotics, defined as non-digestible food components that favorably affect the subject by selectively stimulating the growth and / or activity of one or a limited number of bacteria in the gut, thereby bringing about an improvement in the health of the host.

[0006] Upper gastrointestinal (GI) disorders and / or diseases affect many people worldwide and lead to recurrent and uncomfortable symptoms. For example, upper GI disorders and / or diseases can include heartburn, regurgitation, gastroesophageal reflux disease (GERD), gastritis, and indigestion.

[0007] Many drugs have been developed to treat upper GI diseases, but none are without drawbacks.

[0008] Treatment of the upper GI tract condition can include alkalis and acid suppressants and / or specific dietary changes to certain foods. Antacids, such as aluminum hydroxide and magnesium hydroxide, are generally used to neutralize the acidity of the stomach, resulting in an increase in the pH of the stomach and duodenum. Antacids are immediate-acting but only last for a short period and do not necessarily prevent heartburn when taken before foods or drinks that can cause symptoms.

[0009] H2 receptor antagonists (also known as H2 blockers) are prescription drugs that can also be used to treat upper GI conditions by inhibiting the action of histamine on parietal cells, which in turn inhibits acid secretion. Examples of H2 receptor antagonists include ranitidine hydrochloride, cimetidine, nizatidine, rabeprazole, and famotidine. H2 receptor antagonists are effective, but there is a delay between ingestion and relief from symptoms, and the drugs are expensive.

[0010] Proton pump inhibitors ("PPIs") are another alternative type of drug for treating upper GI diseases. PPIs are prescription drugs that target the amount of gastric acid produced by gastric acid-producing cells. Drugs include omeprazole, omeprazole IR, pantoprazole, lansoprazole, and esomeprazole. PPIs have demonstrated a delay between ingestion and relief from symptoms and are expensive.

[0011] Respiratory disorders and / or diseases typically include cough, sneezing, headache, congestion, sore throat, nasal congestion, rhinitis, and fever, etc. The medicaments include liquid elixirs, cough syrups, cold and influenza capsules, cold and influenza tablets, allergy tablets, effervescent tablets, mouth sprays and nasal drops, cough drops, etc.

[0012] The most commonly used medicaments for treating respiratory disorders and / or diseases are taken or administered orally to address the onset of respiratory symptoms and / or to treat the onset. The products typically contain one or more active substances dissolved or dispersed in a carrier system for oral ingestion or oral delivery into the bloodstream. Typically, respiratory medicaments are respiratory products in the form of cough drops, liquids, or capsules, powders, and effervescent tablets. Most medicaments are relatively inexpensive, but on the other hand, their effects only last for a limited period. Some treatments use a combination of antiviral and anti-inflammatory compounds. Again, the effects of these medicaments are for a limited period, and the inclusion of antiviral agents greatly increases the cost for these types of therapies.

[0013] Elite athletes, such as rugby union players, follow a physiologically and psychologically demanding training schedule with frequent competitive matches, limited recovery, and regular international travel. Collectively, these stress factors can impair immunity and increase the risk of acute upper respiratory symptoms (URS) (e.g., cough, sneeze, sore throat & nasal congestion) and gastrointestinal symptoms (GIS) (e.g., bloating, flatulence, abdominal distension, nausea & diarrhea) (Peters & Bateman, 1983; Drew et al., 2017; Hellard et al., 2015; Svendsen et al., 2016; Wentz et al., 2018). On average, elite rugby union players experience four episodes of upper respiratory illness and one episode of GI complaint per season (Cunniffe et al., 2009), with the highest incidence reported during pre-season and winter (Cunniffe et al., 2009; Tieman et al., 2020; Keaney et al., 2021). Therefore, identifying ways to reduce these illnesses or accelerate recovery to enable return to play is essential for player health and team performance. The profile, genetic material, and functional activity of the gut microbiota (gut microbiome) have important effects on systemic immune function (Roberfoid et al., 2010). Manipulation of the gut microbiome is possible through dietary interventions, most commonly through the ingestion of probiotics or prebiotic nutritional supplements. Previous studies using single or multiple strains of 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). Furthermore, a 2015 Cochrane review reported that probiotics reduce the number and duration of URS episodes in adults and children (Hao et al., 2015).Multiple strains of probiotics reduced the incidence and duration of URS by about two days in elite rugby union players (Haywood et al., 2013). However, the various probiotic strains used across different studies raise uncertainty about exactly which are most beneficial for the health of athletes.

[0014] GOS has also previously been shown to reduce the number of URS days and the severity of GIS in student cohorts (Hughes et al., 2011) and cohorts suffering from traveler's diarrhea (Drakoularakou et al., 2010; Hasle et al., 2017).

[0015] One group of compounds classified as prebiotics is galactooligosaccharides, which are galactose-containing oligosaccharides produced from lactose by the transgalactosylase activity of the β-galactosidase enzyme. Known galactooligosaccharide prebiotic products contain mixtures of many oligosaccharide compounds in various ratios, and not all of them necessarily have a desirable beneficial effect on the consumer's gut microbiota. Such products have shown some beneficial results in improving the health of patients, but there is still a need for further improved oligosaccharide compositions to fully realize the potential benefits for consumers of such prebiotics, as well as to provide a long-lasting and inexpensive treatment for URS and GI conditions and / or diseases, and / or to improve the performance of athletes. SUMMARY OF THE INVENTION

[0016] In particular, it is an object of the present invention to provide a composition comprising an oligosaccharide compound that addresses at least one disadvantage of the prior art, whether specified herein or elsewhere, or to provide an alternative to an existing composition. For example, it may be an object of the present invention to provide a composition comprising an oligosaccharide compound that contains a higher proportion of certain beneficial oligosaccharides than known oligosaccharide compositions.

[0017] According to an aspect of the present invention, there is provided a composition, method and use as described in the appended claims. Other features of the present invention will be apparent from the dependent claims and the following description.

Mode for Carrying Out the Invention

[0018] According to a first aspect of the present invention, there is provided a composition comprising an oligosaccharide compound for use in the prevention, amelioration or treatment of the state of URS and GI and / or a disease, wherein the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c comprising, wherein X a , X b and X c are each independently selected from monosaccharides.

[0019] In an embodiment related to the first aspect of the present invention, there is also provided a method of such prevention, amelioration or treatment, including administration of a composition comprising an oligosaccharide compound to a subject in need of prevention, amelioration or treatment of the state of URS and GI and / or a disease, wherein the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c comprising, wherein X a , X b and X c are each independently selected from monosaccharides.

[0020] In a further embodiment related to the first aspect of the present invention, there is further provided a method of preventing, ameliorating or treating such URS and GI conditions and / or diseases in a subject in need thereof, the method comprising: i) determining whether the subject has URS and GI conditions and / or a disease, and ii) administering to the subject a therapeutically effective amount of a composition comprising an oligosaccharide compound wherein the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c comprising, wherein X a , X b and X c are each independently selected from monosaccharides.

[0021] In still a further embodiment of the present invention, there is provided the use of a composition for the manufacture of a medicament for the prevention, amelioration or treatment of URS and GI conditions and / or diseases, wherein the composition comprises an oligosaccharide compound, and the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) At least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c wherein Xa, Xb and Xc are each independently selected from monosaccharides.

[0022] In a further embodiment of the invention, a performance enhancing composition for a sports person comprising an oligosaccharide compound is provided, wherein the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c wherein X a , X b and X c are each independently selected from monosaccharides.

[0023] In a further embodiment, a composition for use in modulating the URS and GI environment is provided, wherein the composition comprises an oligosaccharide compound, and the oligosaccharide compound is: based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c wherein X a , X b and X c are each independently selected from monosaccharides.

[0024] As used herein, the terms "treatment", "treating", "treat", etc. refer to obtaining the desired pharmacological and / or physiological effect. The effect can be prophylactic in terms of completely or partially preventing a disease or its symptoms, and / or therapeutic in terms of partial or complete cure of the disease and / or side effects caused by the disease. "Treatment" as used herein covers any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing a disease from occurring in a subject who may be susceptible to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., preventing or delaying its progression; and (c) alleviating the disease and / or its symptoms, i.e., causing regression of the disease.

[0025] 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, etc.

[0026] "Therapeutically effective amount" refers to the amount of a compound that is sufficient to effect such treatment of a disease when administered to a subject for treating the disease. A "therapeutically effective amount" will vary depending on the compound(s) used, the disease and its severity, and the age, weight, etc. of the subject to be treated.

[0027] The term "GI condition and / or disease" refers to a series of conditions and / or diseases associated with the gastrointestinal tract having identifiable symptoms, including but not limited to the following upper GI conditions and / or diseases: indigestion, constipation, dysphagia, heartburn, acid reflux, reflux esophagitis, gastroesophageal reflux disease, gastritis, small intestinal bacterial overgrowth and dyspepsia; and the following lower (or more general) GI conditions and / or diseases: constipation, diarrhea, irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), hemorrhoids, anal fissure, perianal abscess, fistula-in-ano, diverticulosis or diverticulitis, colitis, infectious colitis, ulcerative colitis, Crohn's disease, ischemic colitis, radiation colitis, pseudomembranous colitis, Clostridioides difficile colitis, proctocolitis, ameboma, angismus, peptic ulcer, colic, biliary colic, gastroenteritis, enteropathic acrodermatitis, intestinal obstruction, intussusception, polyps, Hirschsprung's disease and diabetes.

[0028] Preferably, the GI condition and / or disease is an upper GI condition and / or disease.

[0029] The term "upper respiratory tract condition and / or disease" or "URS condition and / or disease" refers to a series of conditions and / or diseases associated with the upper respiratory tract having identifiable symptoms, including but not limited to the following respiratory conditions and / or diseases: bacterial infection, viral infection, cold, influenza (flu), respiratory allergy and asthma.

[0030] Preferably, the URS condition and / or disease is an upper respiratory tract infection (URTI).

[0031] URS or GI conditions and / or diseases can be in athletes and / or individuals who are undergoing (or about to undergo) physical exercise or physical training. The composition can be administered before, during, or after exercise or physical activity. Alternatively, the composition can be administered according to a continuous dosing schedule, such as once or twice a day. The therapeutically effective amount can be a standardized dose or a dose determined by individual factors such as the age, body mass index (BMI), or overall body weight of the individual.

[0032] The term "modulating" or "modulates" refers to the effect of beneficially adjusting the GI system or URS environment to make it stronger and more resilient, thereby reducing the disease state or the severity of such a condition in turn.

[0033] The term "performance" refers to the ability of an individual to perform specific physical tasks related to sports and exercise, including but not limited to reducing fatigue and improving endurance.

[0034] The term "sports person" refers to an individual who is actively engaged in sports or exercise, including both professional and amateur individuals.

[0035] The inventors have advantageously found that supplementation with the compositions of the present invention reduces the incidence and severity of URS episodes, GIS, and particularly upper GIS, and increases the sIgA secretion rate in athletes. The composition acts systemically, and studies have shown that the compositions of the present invention can modulate immune function to reduce disease, which improves the health status of athletes and their ability to train and compete.

[0036] These types of oligosaccharides having a specific linkage between galactose units (designated Gal) and terminal monosaccharides are considered to be particularly beneficial for the maintenance of a healthy human gut microbiota and thus beneficial to the health of a subject. Known oligosaccharide compositions either do not contain each of these types of oligosaccharides or contain lower levels of these oligosaccharides. Accordingly, the compositions of this first aspect provide an advantage to consumers in the establishment and maintenance of the human gut microbiota when used as prebiotics. Advantageously and unexpectedly, the compositions of the present invention have been found to be more actively consumed by the microbiota present in the gut when compared to other oligosaccharides, and during human trials, the compositions have also been shown to significantly reduce URS and GI symptoms.

[0037] The weight percentages of the specific oligosaccharide compounds discussed herein are based on the total weight of all the oligosaccharide compounds present in the total composition. Accordingly, when determining a specific weight percentage, only the fraction of the composition provided by the oligosaccharide, which can be any of components (a), (b), (c) or any other oligosaccharide present, is considered. For this determination of oligosaccharide content, disaccharides are included, apart from lactose. The compositions of this first aspect may contain other non-oligosaccharide components, including monosaccharides and lactose. These components are not considered when determining the specific weight percentages of the oligosaccharide compounds discussed herein. The specific amount of oligosaccharide compound in the composition can be referred to as the weight percentage of the oligosaccharide fraction of the composition of this first aspect.

[0038] The oligosaccharide compounds (a), (b) and (c) each independently comprise an X selected from sugars a , X b and X c respectively. The X a , X b and X c groups can be considered to be sugar units. Accordingly, the groups X a , X b and X ccan be considered as the terminal sugar of the oligosaccharide compound. Group X a , X b and X c are preferably independently selected from monosaccharides. The galactose units of compounds (a), (b) and (c) and any suitable monosaccharide units capable of forming an oligosaccharide can provide X a , X b and X c . Preferably X a , X b and X c are each independently selected from the following monosaccharides. Suitable monosaccharide units are 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 mixtures thereof. Thus, each of compounds (a), (b) and (c) can contain a mixture of oligosaccharide compounds having different respective X groups, for example either a Glc or Fuc X group.

[0039] In some embodiments, groups X a , X b and X c are independently selected from the monosaccharides listed above and are preferably independently selected from Glc, Fuc, Ara, Xyl, Rha and Man or mixtures thereof.

[0040] The sugar units of the oligosaccharide in the composition of this first aspect can have either the D enantiomeric form or the L enantiomeric form. Preferably, all of the Gal sugar units in components (a), (b) and (c) have the D enantiomeric form. Components (a), (b) and (c) can thus be as follows: (a) D-Gal-(β1-3)-D-Gal-(β1-4)-X a ; (b) D-Gal-(β1-3)-D-Gal-(β1-3)-X b ; and (c) D-Gal-(β1-3)-D-Gal-(β1-2)-X c .

[0041] Preferably, both Gal and X a , X b and X c sugar units all have the D-configuration. Components (a), (b) and (c) can thus be as follows: (a) D-Gal-(β1-3)-D-Gal-(β1-4)-D-X a ; (b) D-Gal-(β1-3)-D-Gal-(β1-3)-D-X b ; and (c) D-Gal-(β1-3)-D-Gal-(β1-2)-D-X c .

[0042] In some embodiments, each of X a , X b and X c is Glc. Accordingly, compounds (a), (b) and (c) can be galactooligosaccharide compounds (GOS), and the compositions of this first aspect can be referred to as galactooligosaccharide compositions. Such galactooligosaccharides can be formed by converting lactose to the stated oligosaccharides using a suitable galactosidase enzyme. In such embodiments, components (a), (b) and (c) are preferably as follows: (a) Gal-(β1-3)-Gal-(β1-4)-Glc; (b) Gal-(β1-3)-Gal-(β1-3)-Glc; and (c) Gal-(β1-3)-Gal-(β1-2)-Glc.

[0043] Preferably, all of the Gal and Glc sugar units have the D-enantiomeric form. Components (a), (b), and (c) can thus be as follows: (a) D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glc; (b) D-Gal-(β1-3)-D-Gal-(β1-3)-D-Glc; and (c) D-Gal-(β1-3)-D-Gal-(β1-2)-D-Glc.

[0044] In some embodiments, each of X a , X b and X c is a mixture of Glc and one or more of the other sugar units described above, such as Fuc, Ara, Xyl, Rha, and Man. Thus, each of (a), (b), and (c) can include a mixture of oligosaccharide compounds having either Glc or one of the other sugar units described above as the X group. In such embodiments, components (a), (b), and (c) can be formed by converting a mixture of lactose and a suitable additional sugar, such as fucose, arabinose, xylose, rhamnose, and mannose, to an oligosaccharide. Preferably, each of X a , X b and X c is a mixture of Glc and Fuc.

[0045] Other oligosaccharides in the composition containing the oligosaccharide compound of this first aspect can, in addition to components (a), (b), and (c) discussed above, also include the sugar unit "X" groups mentioned above.

[0046] The composition of this first aspect contains (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a . Preferably, the composition contains at least 9% by weight of component (a) or at least 10% by weight of component (a).

[0047] Preferably, the composition comprises up to 35% by weight of component (a), up to 30% by weight of component (a) or up to 25% by weight of component (a).

[0048] Preferably, the composition comprises from 8 to 35% by weight of component (a), from 8 to 25% by weight of component (a) or from 10 to 20% by weight of component (a).

[0049] The composition of this first aspect comprises (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b Preferably, the composition comprises at least 4% by weight of component (b) or at least 5% by weight of component (b).

[0050] Preferably, the composition comprises up to 25% by weight of component (b), up to 20% by weight of component (b) or up to 15% by weight of component (b).

[0051] Preferably, the composition comprises from 3 to 25% by weight of component (b), from 4 to 20% by weight of component (b) or from 4 to 10% by weight of component (b).

[0052] The composition of this first aspect comprises (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c Preferably, the composition comprises at least 6% by weight of component (c).

[0053] Preferably, the composition comprises up to 25% by weight of component (c), up to 20% by weight of component (c) or up to 15% by weight of component (c).

[0054] Preferably, the composition comprises from 5 to 25% by weight of component (c), from 5 to 20% by weight of component (c) or from 6 to 10% by weight of component (c).

[0055] The above amounts are based on the total weight of the oligosaccharide compounds present in the composition.

[0056] Preferably, in the composition of this first aspect: Based on the total weight of the oligosaccharide compounds present in the composition, component (a) is present in an amount of up to 35% by weight; component (b) is present in an amount of up to 25% by weight; and component (c) is present in an amount of up to 25% by weight.

[0057] Preferably, in the composition of this first aspect: Based on the total weight of the oligosaccharide compounds present in the composition, component (a) is present in an amount of 8 to 25% by weight; component (b) is present in an amount of 3 to 25% by weight; and component (c) is present in an amount of 5 to 20% by weight.

[0058] In some embodiments, the composition of this first aspect is: Based on the total weight of the oligosaccharide compounds present in the composition, (a) 8 to 25% by weight of Gal-(β1-3)-Gal-(β1-4)-Glc; (b) 3 to 25% by weight of Gal-(β1-3)-Gal-(β1-3)-Glc; and (c) 5 to 20% by weight of Gal-(β1-3)-Gal-(β1-2)-Glc comprising.

[0059] Preferably, the weight percentage ratio of compound (a) to compound (b) is from 1:1 to 3:1, preferably from 1.5:1 to 2.5:1.

[0060] Preferably, the weight percentage ratio of compound (a) to compound (c) is from 1:1 to 3:1, preferably from 1.5:1 to 2.5:1.

[0061] Preferably, the weight percentage ratio of compound (b) to compound (c) is from 2:1 to 1:2, preferably from 1.5:1 to 1:1.5.

[0062] Preferably, in the composition of this first aspect, the oligosaccharide compound is: Based on the total weight of the oligosaccharide compounds present in the composition, (d) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-X d wherein X d is selected from monosaccharides.

[0063] X d is X a X b and X c can be selected from the same monosaccharide units as described above for X

[0064] In some embodiments, X d is Glc.

[0065] Preferably, the Gal sugar units of component (d) have the D enantiomeric form. Preferably, all of the Gal and X d sugar units have the D enantiomeric form. Component (d) can thus be as follows: D-Gal-(β1-3)-D-Gal-(β1-3)-D-Gal-(β1-4)-D-X d .

[0066] In some embodiments, X d is a mixture of Glc and one or more other monosaccharide units, such as Fuc, Ara, Xyl, Rha and Man, preferably Fuc.

[0067] Preferably, the composition comprises at least 4% by weight of component (d) or at least 5% by weight of component (d).

[0068] Preferably, the composition comprises up to 25% by weight of component (d), up to 20% by weight of component (d) or up to 15% by weight of component (d).

[0069] Preferably, the composition comprises from 3 to 25% by weight of component (d), from 4 to 20% by weight of component (d) or from 4 to 10% by weight of component (d).

[0070] Preferably, in the composition of this first aspect: Based on the total weight of the oligosaccharide compounds present in the composition, Component (a) is present in an amount of 8 to 25% by weight; Component (b) is present in an amount of 3 to 25% by weight; Component (c) is present in an amount of 5 to 20% by weight; and Component (d) is present in an amount of 3 to 25% by weight.

[0071] In some embodiments, the composition of this first aspect is: Based on the total weight of the oligosaccharide compounds present in the composition, (a) 8 to 25% by weight of Gal-(β1-3)-Gal-(β1-4)-Glc; (b) 3 to 25% by weight of Gal-(β1-3)-Gal-(β1-3)-Glc; (c) 5 to 20% by weight of Gal-(β1-3)-Gal-(β1-2)-Glc; and (d) 3 to 25% by weight of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-Glc comprising.

[0072] Preferably, in the composition of this first aspect, the oligosaccharide compound is: Based on the total weight of the oligosaccharide compounds present in the composition, (e) at least 5% by weight of Gal-(β1-4)-Gal-(β1-4)-X e comprising, wherein X e is selected from monosaccharides.

[0073] X e is X a , X b and X c can be selected from the same monosaccharide units as described above for X

[0074] Preferably, the Gal sugar unit of component (e) has the D enantiomeric form. Preferably, Gal and X eAll sugar units have the D-enantiomeric form. Component (e) can thus be as follows: D-Gal-(β1-4)-D-Gal-(β1-4)-D-X e .

[0075] In some embodiments, X e is Glc.

[0076] In some embodiments, X e is a mixture of Glc and one or more other monosaccharide units, such as Fuc, Ara, Xyl, Rha and Man, preferably Fuc.

[0077] Preferably, the composition contains at least 6% by weight of component (e) or at least 7% by weight of component (e).

[0078] Preferably, the composition contains at most 25% by weight of component (e), at most 20% by weight of component (e) or at most 15% by weight of component (e).

[0079] Preferably, the composition contains 5 to 25% by weight of component (e), 5 to 20% by weight of component (e) or 6 or 10% by weight of component (e).

[0080] Preferably, in the composition of this first aspect, based on the total weight of the oligosaccharide compounds present in the composition, component (a) is present in an amount of 8 to 25% by weight; component (b) is present in an amount of 3 to 25% by weight; component (c) is present in an amount of 5 to 20% by weight; component (d) is present in an amount of 3 to 25% by weight; and component (e) is present in an amount of 5 to 25% by weight.

[0081] In some embodiments, the composition of this first aspect is: based on the total weight of the oligosaccharide compounds present in the composition, (a) 8 to 25% by weight of Gal-(β1-3)-Gal-(β1-4)-Glc; (b) 3 to 25% by weight of Gal-(β1-3)-Gal-(β1-3)-Glc; (c) 5 to 20% by weight of Gal-(β1-3)-Gal-(β1-2)-Glc; and (d) 3 to 25% by weight of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-Glc; and (e) 5 to 25% by weight of Gal-(β1-4)-Gal-(β1-4)-Glc comprising.

[0082] In the composition of this first aspect, X a , X b , X c , X d and X e are each independently selected from the above monosaccharides.

[0083] In the composition of this first aspect, X a , X b , X c , X d and X e can each independently be selected from Glc, Fuc, Ara, Xyl, Rha and Man or mixtures thereof.

[0084] In some embodiments, X a , X b , X c , X d and X e are each Glc.

[0085] In some embodiments, X a , X b , X c , X d and X e each contain Fuc. Preferably, X a , X b , X c , X d and X eThey are each a mixture of Glc and Fuc.

[0086] The oligosaccharide compounds of the composition of this first aspect preferably have a relatively high ratio of β1-3 Gal-Gal linkages, mainly due to the presence of components (a), (b), (c) and optionally (d) in the ratios discussed herein. The inventors have found that such a relatively high ratio of β1-3 Gal-Gal linkages can be particularly advantageous for the use of the compositions discussed herein.

[0087] Preferably, 35 to 55% of the Gal-Gal linkages in the oligosaccharide compound are 1-3 linkages, preferably 40 to 55%, preferably 40 to 50%.

[0088] Preferably, 40 to 60% of the Gal-Gal linkages in the oligosaccharide compound are 1-4 linkages, preferably 45 to 55%, preferably 45 to 52%.

[0089] Preferably, 8 to 20% of the Gal-X linkages in the oligosaccharide compound are 1-3 linkages, preferably 10 to 18%, preferably 12 to 16%, and preferably here, X is Glc.

[0090] Preferably, 15 to 25% of the Gal-X linkages in the oligosaccharide compound are 1-4 linkages, preferably 16 to 24%, preferably 17 to 22%, and preferably here, X is Glc.

[0091] Preferably, 30 to 45% of the Gal-X linkages in the oligosaccharide compound are 1-2 linkages, preferably 32 to 43%, preferably 34 to 41%, and preferably here, X is Glc.

[0092] Preferably, 20 to 36% of the Gal-X linkages in the oligosaccharide compound are 1-6 linkages, preferably 22 to 34%, preferably 25 to 32%, and preferably here, X is Glc.

[0093] Preferably, the Gal-X mentioned above, the bond, for example the Gal-Glu bond, is a β-linkage, i.e., a β-glycosidic bond.

[0094] Preferably, the composition contains at least 25% by weight of trisaccharide, based on the total weight of the composition. Preferably, the composition contains at least 28% by weight of trisaccharide or at least 30% by weight of trisaccharide.

[0095] Preferably, the composition contains a maximum of 70% by weight of trisaccharide, preferably a maximum of 60% by weight of trisaccharide or a maximum of 50% by weight of trisaccharide.

[0096] Preferably, the composition contains 25 to 70% by weight of trisaccharide, preferably 30 to 60% by weight of trisaccharide or 30 to 50% by weight of trisaccharide.

[0097] Preferably, the composition contains at least 10% by weight of tetrasaccharide, based on the total weight of the composition. Preferably, the composition contains at least 12% by weight of tetrasaccharide or at least 15% by weight of tetrasaccharide. Preferably, the composition contains at least 20% by weight of tetrasaccharide or at least 25% by weight of tetrasaccharide.

[0098] Preferably, the composition contains a maximum of 40% by weight of tetrasaccharide, preferably a maximum of 25% by weight of tetrasaccharide or a maximum of 20% by weight of tetrasaccharide.

[0099] Preferably, the composition contains 10 to 40% by weight of tetrasaccharide, preferably 10 to 30% by weight of tetrasaccharide or 12 to 25% by weight of tetrasaccharide.

[0100] Preferably, the composition contains 30 to 50% by weight of trisaccharide and 10 to 25% by weight of tetrasaccharide, based on the total weight of the composition.

[0101] As described above, the determination of the disaccharides present in the composition excludes lactose. Thus, in some embodiments, the composition preferably contains up to 40% by weight of disaccharides, preferably up to 30% by weight of disaccharides or up to 20% by weight of disaccharides. Preferably, the lactose content in the composition of this first aspect is minimized. Preferably, the composition is substantially free of lactose. Preferably, the composition does not contain lactose.

[0102] Preferably, the composition contains 10 to 40% by weight of disaccharides, preferably 10 to 30% by weight of disaccharides or 10 to 20% by weight of disaccharides.

[0103] Preferably, the composition contains 10 to 40% by weight of disaccharides, 30 to 60% by weight of trisaccharides and 10 to 25% by weight of tetrasaccharides, based on the total weight of the composition.

[0104] The following description relates to the content of disaccharides in the oligosaccharide compound. Preferably, the oligosaccharide compound contains up to 40% by weight of disaccharides, preferably up to 30% by weight of disaccharides or up to 20% by weight of disaccharides.

[0105] Preferably, the oligosaccharide compound contains 0 to 40% by weight of disaccharides, preferably 10 to 30% by weight of disaccharides or 10 to 20% by weight of disaccharides.

[0106] Preferably, the oligosaccharide compound contains at least 25% by weight of trisaccharides, based on the total weight of the oligosaccharide compound present in the composition. Preferably, the oligosaccharide compound contains at least 30% by weight of trisaccharides or at least 33% by weight of trisaccharides.

[0107] Preferably, the oligosaccharide compound contains up to 75% by weight of trisaccharides, preferably up to 65% by weight of trisaccharides or up to 55% by weight of trisaccharides.

[0108] Preferably, the oligosaccharide compound contains 25 to 75% by weight of trisaccharides, preferably 30 to 65% by weight of trisaccharides or 34 to 55% by weight of trisaccharides.

[0109] Preferably, the oligosaccharide compound contains at least 10% by weight of tetrasaccharide based on the total weight of the oligosaccharide compound present in the composition. Preferably, the oligosaccharide compound contains at least 12% by weight of tetrasaccharide or at least 15% by weight of tetrasaccharide.

[0110] Preferably, the oligosaccharide compound contains a maximum of 45% by weight of tetrasaccharide, preferably a maximum of 35% by weight of tetrasaccharide or a maximum of 30% by weight of tetrasaccharide.

[0111] Preferably, the oligosaccharide compound contains 10 to 45% by weight of tetrasaccharide, preferably 10 to 35% by weight of tetrasaccharide or 15 to 30% by weight of tetrasaccharide.

[0112] Preferably, the oligosaccharide compound contains 0 to 40% by weight of disaccharide and 30 to 75% by weight of trisaccharide based on the total weight of the oligosaccharide compound present in the composition.

[0113] Preferably, the oligosaccharide compound contains 0 to 40% by weight of disaccharide, 30 to 75% by weight of trisaccharide and 10 to 45% by weight of tetrasaccharide based on the total weight of the oligosaccharide compound present in the composition.

[0114] The composition of this first aspect preferably contains at least 50% by weight of oligosaccharide compound, including components (a), (b), (c) and optionally (d) and (e). Preferably, the composition contains at least 55% by weight of oligosaccharide compound, preferably at least 60% by weight, based on the total weight of the composition.

[0115] Preferably, the composition contains a maximum of 100% by weight of oligosaccharide compound, preferably a maximum of 95%, a maximum of 90% or a maximum of 85% of oligosaccharide compound.

[0116] Preferably, the composition contains 50 to 100% by weight of oligosaccharide compound, preferably 55 to 95%, or 60 to 85% of oligosaccharide compound.

[0117] In some embodiments, the composition is in the form of a syrup. The syrup preferably contains at least 50% by weight of the oligosaccharide compound, at least 55% by weight of the oligosaccharide compound, or at least 60% by weight. Preferably, the syrup contains 50 to 75% by weight of the oligosaccharide compound, preferably 55 to 70% or 60 to 70% by weight of the oligosaccharide compound.

[0118] The syrup may contain a significant amount of monosaccharides, such as glucose and / or galactose. The syrup may contain 15 to 30% by weight of monosaccharides, preferably 20 to 28% by weight of monosaccharides or 21 to 25% by weight of monosaccharides, such as glucose and / or galactose.

[0119] The syrup preferably contains 20 to 30% by weight of water, preferably 22 to 28% by weight of water.

[0120] The syrup may also contain lactose, for example 4 to 14% by weight of lactose.

[0121] In some embodiments, the composition is in the form of a powder. The powder preferably contains at least 60% by weight of the oligosaccharide compound, preferably at least 70% or at least 75% by weight of the oligosaccharide compound. Preferably, the powder contains 60 to 100% by weight of the oligosaccharide compound, preferably 70 to 95% or 75 to 90% by weight of the oligosaccharide compound.

[0122] The powder preferably contains a reduced amount of monosaccharides, such as glucose and / or galactose, compared to the syrup discussed above. The powder may contain 1 to 10% by weight of monosaccharides, preferably 2 to 8% by weight of monosaccharides or 3 to 7% by weight of monosaccharides, preferably about 5% by weight, such as glucose and / or galactose.

[0123] The powder preferably contains 1 to 10% by weight of water, preferably 3 to 6% by weight of water.

[0124] The composition of this first aspect may be purified to remove monosaccharides and optionally disaccharides from the composition.

[0125] The composition of this first aspect may be fractionated to separate the oligosaccharide component of the composition according to their molecular weights, for example to remove disaccharides from the composition or to isolate 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 such fractionation may be referred to as an oligosaccharide fraction or a GOS fraction.

[0126] In such embodiments, the composition (or oligosaccharide fraction) preferably comprises at least 70% by weight of trisaccharides, tetrasaccharides and higher oligosaccharides, preferably at least 80% by weight or at least 90% by weight. Such higher oligosaccharides have a degree of polymerization of 5 and above. Preferably, the composition comprises at least 95% by weight of trisaccharides, tetrasaccharides and higher oligosaccharides. Preferably, the composition consists of or consists essentially of trisaccharides, tetrasaccharides and higher oligosaccharides.

[0127] In such embodiments, the composition preferably comprises 40 to 70% by weight of trisaccharides, preferably 45 to 70% by weight of trisaccharides or 50 to 70% by weight of trisaccharides.

[0128] Preferably, the oligosaccharide compound comprises 15 to 50% by weight of tetrasaccharides, preferably 15 to 40% by weight of tetrasaccharides or 20 to 40% by weight of tetrasaccharides.

[0129] Preferably, the oligosaccharide compound comprises 5 to 25% by weight of higher oligosaccharides, preferably 5 to 20% by weight of higher oligosaccharides or 10 to 20% by weight of higher oligosaccharides.

[0130] Preferably, the composition comprises, based on the total weight of the composition, 40 to 70% by weight of trisaccharides, 15 to 40% by weight of tetrasaccharides and 5 to 25% by weight of higher oligosaccharides. Preferably, the composition comprises, based on the total weight of the composition, 50 to 70% by weight of trisaccharides, 20 to 40% by weight of tetrasaccharides and 10 to 20% by weight of higher oligosaccharides.

[0131] In such embodiments, the composition (or oligosaccharide fraction) preferably comprises the above components (a), (b) and (c) in the following amounts: Based on the total weight of the oligosaccharide compounds present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c (wherein X a , X b and X c are each independently selected from monosaccharides).

[0132] Preferably, the composition (or oligosaccharide fraction) comprises the above components (a), (b) and (c) in the following amounts: (a) at least 10% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 5% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 7% by weight of Gal-(β1-3)-Gal-(β1-2)-X c .

[0133] The composition may comprise the above component (d) and / or component (e).

[0134] The composition preferably comprises the said components in the ratios discussed above.

[0135] In some embodiments, the composition of this first aspect is a fractionated product of trisaccharides and tetrasaccharides (which can be referred to as the DP3 / DP4 fraction).

[0136] In such embodiments, the composition (or DP3 / DP4 fraction) preferably contains at least 70% by weight of trisaccharides and tetrasaccharides, preferably at least 80% by weight or at least 90% by weight. Preferably, the composition consists of or consists essentially of trisaccharides and tetrasaccharides.

[0137] In such embodiments, the composition preferably contains 50 to 80% by weight of trisaccharides, preferably 55 to 75% by weight of trisaccharides or 60 to 75% by weight of trisaccharides.

[0138] Preferably, the oligosaccharide compound contains 20 to 50% by weight of tetrasaccharides, preferably 25 to 45% by weight of tetrasaccharides or 25 to 40% by weight of tetrasaccharides.

[0139] Preferably, the composition contains 50 to 80% by weight of trisaccharides and 20 to 50% by weight of tetrasaccharides based on the total weight of the composition. Preferably, the composition contains 60 to 75% by weight of trisaccharides and 25 to 40% by weight of tetrasaccharides based on the total weight of the composition.

[0140] In such embodiments, the composition (or oligosaccharide fraction) preferably contains the above components (a), (b) and (c) in the following amounts: Based on the total weight of the oligosaccharide compounds present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c (wherein X a , X b and X care each independently selected from monosaccharides).

[0141] Preferably, the composition (or oligosaccharide fraction) comprises the above components (a), (b) and (c) in the following amounts: (a) at least 10% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 5% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 7% by weight of Gal-(β1-3)-Gal-(β1-2)-X c .

[0142] The composition may comprise the above component (d) and / or component (e).

[0143] The composition preferably comprises the said components in the ratios discussed above.

[0144] In some embodiments, the composition of this first aspect is a trisaccharide fraction product (which may be referred to as the DP3 fraction). Such a trisaccharide fraction product can be obtained by the known fractionation methods mentioned above. Such a composition preferably comprises at least 70% by weight of trisaccharide, preferably at least 80% by weight or at least 90% by weight. Preferably, the composition comprises at least 95% by weight of trisaccharide.

[0145] In such embodiments, the composition (or oligosaccharide fraction) preferably comprises the above components (a), (b) and (c) in the following amounts: Based on the total weight of the oligosaccharide compounds present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c (wherein Xa and X b and X c are each independently selected from monosaccharides).

[0146] Preferably, the composition (or oligosaccharide fraction) comprises the above components (a), (b) and (c) in the following amounts: (a) at least 10% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 5% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 7% by weight of Gal-(β1-3)-Gal-(β1-2)-X c .

[0147] The composition may comprise the above component (e).

[0148] The composition preferably comprises the said components in the ratios discussed above.

[0149] The composition of this first aspect may be for use as a food supplement product for consumption by consumers and may be incorporated therein. Such products may be selected from the group consisting of dairy products (e.g., liquid milk, dried milk powders such as whole milk powder, skim milk powder, filled milk powder, whey powder, infant formula, ice cream, yogurt, cheese, fermented dairy products), beverages, sports drinks, baby foods, cereals, breads, biscuits, confectionery, cakes, food supplements, dietary supplements, animal feeds, poultry feeds or indeed any other food or beverage.

[0150] The composition of this first aspect may be incorporated into a synbiotics composition. Such a synbiotics composition is preferably a mixture comprising live microorganisms and a substrate (s) selectively utilized by the host microorganisms which confer health benefits on the host, i.e., probiotics and prebiotics.

[0151] The composition of this first aspect can be for use as a pharmaceutical composition or a nutraceutical composition comprising at least one carrier, excipient, or diluent, and can be in that form.

[0152] Suitable further components of the pharmaceutical composition or nutraceutical composition, and methods of preparing such pharmaceutical compositions or nutraceutical compositions are known in the art.

[0153] The composition can be administered in a single dose or in multiple doses. Suitable frequencies of administration can be at least once a day, every other day, once a week, once every two, three, 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, two, three, four, or five years, or over the course of the disease, or over the lifetime of the subject.

[0154] It will be apparent to those skilled in the art that the administration of the composition is optimized during clinical trials.

[0155] The composition of the present invention can be formulated into a pharmaceutical composition by combination with a suitable pharmaceutically acceptable carrier, pharmaceutically acceptable diluent, or pharmaceutically acceptable excipient, and can be formulated into a solid, semi-solid, or liquid form of preparation such as tablets, capsules, powders, granules, and solutions.

[0156] Pharmaceutically acceptable carriers, excipients, or diluents include, for example: water, physiological saline, glucose, maltodextrin, glycerol, ethanol, salts such as NaCl, MgCl2, KCl, MgSO4, etc.; buffers such as phosphate buffer, citrate buffer, Tris buffer, N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), sodium 2-(N-morpholino)ethanesulfonate (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS), etc.; solubilizing agents; detergents such as non-ionic detergents like Tween-20; glycerol; and the like.

[0157] Pharmaceutically acceptable carriers, excipients, and diluents are non-toxic to the recipient at the dosages and concentrations used and include, for example, buffers such as phosphoric acid, citric acid, 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 combinations thereof); amino acids such as arginine, glycine, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, tyrosine, tryptophan, 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 such as trehalose, sucrose, lactose, glucose, mannose, maltose, galactose, fructose, sorbose, raffinose, glucosamine, N-methylglucosamine, galactosamine, and neuraminic acid; and / or non-ionic surfactants such as Tween, Brij Pluronics, Triton-X, or polyethylene glycol (PEG).

[0158] For oral preparations, the compositions of the present invention may include suitable additives for making tablets, powders, granules or capsules, such as 30 conventional additives such as lactose, mannitol, corn starch or potato starch; binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatin; disintegrants such as corn starch, potato starch or sodium carboxymethyl cellulose; lubricants such as talc or magnesium stearate; and, if desired, diluents, buffers, wetting agents, preservatives and flavorings.

[0159] The pharmaceutical composition can be in liquid form, lyophilized form or liquid form reconstituted from lyophilized form (where the lyophilized formulation will be reconstituted with a sterile solution prior to administration). Standard procedures for reconstituting lyophilized compositions involve adding back a large volume of pure water (typically equivalent to the volume removed during lyophilization).

[0160] An isotonic agent can be included in the formulation to adjust the tonicity of the formulation. Exemplary isotonic agents include sodium chloride, potassium chloride, glycerin, and any component from the group of amino acids, saccharides and combinations thereof. In some embodiments, hypertonic or hypotonic solutions may be appropriate, but aqueous formulations are isotonic. The term "isotonic" refers to a solution having the same tonicity as some other solution to which it is compared, such as physiological saline or serum.

[0161] The composition can regulate the abundance of bacterial genera present in the upper GI tract. In some embodiments, the composition regulates the abundance of bacterial genera present in one or both of the small intestine or the large intestine. In some embodiments, the composition regulates the abundance of a dominant bacterial genus in the small intestine selected from the group consisting of Acromobacter, Agrobacterium, Blautia, Burkholderia, Coprococcus, Cryocola, Enterococcus, Eubacterium, Holdemania, Lactococcus, Mycobacterium, Pseudoramibacter, Ralstonia, Sphingomonas, Streptococcus, and Turicibacter.

[0162] 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: (i) providing a source of a sugar compound; (ii) treating the source of the sugar compound with one or more galactosidase enzymes to convert at least partially the source of the sugar compound into an oligosaccharide compound and the steps thereof.

[0163] Preferably, the steps of the method of this second aspect are carried out in the order of step (ii) after step (i).

[0164] Preferably, the source of the sugar compound comprises lactose, lactulose or epilactose. Preferably, the source of the sugar compound comprises lactose. The source of the sugar can be lactose syrup, for example derived from cow's milk. Lactose can be heat treated.

[0165] In some embodiments, no additional sugars, such as monosaccharides or disaccharides, are added to the source of the sugar. In such embodiments, the method produces a galacto-oligosaccharide compound.

[0166] In some embodiments, the source of the sugar compound comprises at least one additional sugar. Preferably, at least one additional sugar provides an oligosaccharide compound having alternative terminal monosaccharide units, as discussed above. Preferably, at least one additional sugar is a source of such monosaccharide units. At least one additional sugar that is a source of such monosaccharide units can be a monosaccharide or a higher sugar such as a disaccharide. At least one additional sugar can be a source of a monosaccharide 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. At least one additional sugar can be one or more of the monosaccharides listed above.

[0167] In such embodiments, the method produces an oligosaccharide having one or more of the above monosaccharides as terminal sugar units.

[0168] In some embodiments, at least one additional sugar can be selected from fucose, arabinose, xylose, rhamnose, mannose or a mixture thereof. The source of the sugar can comprise lactose and a source of one or more of the above monosaccharides. In such embodiments, the method produces an oligosaccharide having a terminal sugar unit selected from Glc, Fuc, Ara, Xyl, Rha and Man, or a mixture thereof.

[0169] Step (ii) of the method involves treating a source of sugar compounds with at least one galactosidase enzyme. The galactosidase enzyme can be an alpha-galactosidase enzyme or a beta-galactosidase enzyme, depending on whether an alpha bond or a beta bond is required between the sugar units of the oligosaccharide compound. Preferably, the enzyme exhibits galactosyltransferase (transgalactoside) activity and forms an alpha bond or a beta bond between the sugar units in the source of sugar compounds. This results in the synthesis of oligosaccharide compounds having two or more galactose units derived from lactose. Preferably, step (ii) is carried out until the conversion of the source of sugar to oligosaccharide compounds is complete.

[0170] Step (ii) can involve treating the source of sugar compounds with one or more additional enzymes other than galactosidase enzymes.

[0171] Preferably, the method includes step (iii) of separating the galactosidase enzyme from the composition containing the oligosaccharide compound. Step (iii) can involve removing the enzyme by filtration, for example, by nanofiltration.

[0172] The composition containing the oligosaccharide compound produced in step (iii) can be heat-treated.

[0173] In some embodiments, as discussed above with respect to the first aspect, the composition is evaporated to reduce the water content and provide the final composition containing the oligosaccharide compound as a syrup.

[0174] In some embodiments, glucose is removed from the composition produced by step (iii) before evaporation. This preferably reduces the glucose content of the composition from 20 - 30 wt% to less than 10 wt%, preferably about 5 wt% or less. As discussed above with respect to the first aspect, the water content of the composition is then reduced by evaporation and the product is dried to provide the final composition containing the oligosaccharide compound as a powder.

[0175] Furthermore, those skilled in the art will recognize that by combining the oligosaccharides obtained and isolated from different sources in the required amounts, a composition containing an oligosaccharide compound according to the first aspect, i.e., a composition containing a specific amount of a specific oligosaccharide compound, can be produced.

Brief Description of the Drawings

[0176] The present invention will be described below by way of example only, with reference to the accompanying drawings:

Figure 1

Figure 2

Figure 3

Figure 4

Examples

[0177] Example 1 - Oligosaccharide Syrup A composition containing an oligosaccharide compound according to the present invention in the form of a syrup was obtained by the following procedure. Lactose was rehydrated with portable water to obtain a working solution of solids between 35 and 65% by weight. The lactose solution was heat-treated and then cooled to 40 - 65°C. The pH of the solution was adjusted to pH 5.5 - 7.5. Beta-galactosidase enzyme was then added to the solution in a sealed container and subsequently reacted with lactose to produce an oligosaccharide compound, catalyzing the transfer of galactose molecules. The progress of the reaction was monitored by measuring the production of glucose. The reaction was allowed to proceed for a period between 8 and 26 hours. The reaction was then stopped by high heat treatment. The reaction mixture was cooled and filtered by carbon filtration to remove the enzyme. The mixture was then dried by evaporation to reduce the water content to about 22 - 28% by weight, obtaining the product as a syrup.

[0178] Example 2 - Oligosaccharide Powder A composition containing an oligosaccharide compound according to the present invention in the form of a powder was obtained by a modification of the above procedure. After removal of the enzyme, the mixture was further filtered to remove a substantial portion of glucose and other monosaccharides, reducing the monosaccharide content from about 23% by weight to about 5% by weight. The water content of the composition was then reduced by evaporation and the product dried to obtain a powder having a water content of up to about 3 - 6% by weight.

[0179] Example 3 - Fractionation Isolation from the sample of Example 2 of different fractions (DP2, DP3, DP4 or DP5) was carried out using a 5×70 cm BioGel P2 column with water as the eluent. The column was operated at a flow rate of 40 to 100 mL / hour 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 in ultrapure water was loaded. This solution was loaded onto the column a total of 15 times to obtain sufficient material of the lower fractions, i.e., DP5. After an exclusion volume of approximately 725 mL, 5 mL fractions were collected. Analysis using thin layer chromatography (TLC) and HPAEC-PAD was performed to ensure that molecules with the same DP were pooled.

[0180] The pooled fractions were frozen and freeze-dried. The dried materials of all runs were combined and redissolved. After the second cycle of freezing and freeze-drying, the dried materials were stored at 4 °C for further analysis or experiments.

[0181] Comparative Example 1 A commercially available composition containing an oligosaccharide compound was obtained in powder form.

[0182] Oligosaccharide analysis The samples of Example 2 and Comparative Example 1 of the present invention were analyzed to determine their oligosaccharide contents by the following procedure.

[0183] Materials and methods Samples of the dry powders of Example 2 and Comparative Example 1 were dissolved in water to obtain solutions having a concentration of 40 g / l for analysis.

[0184] Gel permeation chromatography An HPLC apparatus equipped with Rezex RSO and an RI detector and in-line desalting (for removal of charged substances such as salts and proteins) was used for aqueous GPC separation of the components of the samples. The separation was carried out at a high temperature (80 °C). The separation range of the Rezex RSO column is from DP1 (monosaccharide) to about DP10. All samples were analyzed without dilution (at 40 g / L). Before analysis, all sample solutions were treated at 100 °C for 10 minutes to remove any microbial or enzymatic activity.

[0185] GOS fingerprinting HPAEC-PAD (High-Performance Anion-Exchange Chromatography) equipped with a PA-1 column was used for the separation of monosaccharides and oligosaccharides in various samples. Efforts were made to achieve the separation quality described in van Leeuwen et al., Carbohydrate Research 2016, 425, 48-58. A commercially available malt oligosaccharide mixture and Comparative Example 1 were also injected for comparison with those reported by van Leeuwen et al. in the chromatogram. Based on this, peak annotation was performed for many peaks. The samples used for GPC were diluted 100-fold with DMSO before injection.

[0186] Results Gel Permeation Chromatography (GPC-RI) Table 1 shows the DP (degree of polymerization) composition results for the samples using a Rezex-RSO system based on RI calibration with glucose (values are expressed as g / L in the sample). All materials eluting in the DP5 window were combined.

[0187]

Table 1

[0188] As shown in Table 1, using the concentration information, the relative weight percentages of different DP fractions of oligosaccharides contained in the sample can be calculated, where DP = 2 refers to disaccharides, DP = 3 refers to trisaccharides, and so on.

[0189] GOS Fingerprinting To identify the individual galacto-oligosaccharides in the sample (GOS fingerprinting), a gradient was developed that gave a separation comparable to that reported in van Leeuwen et al., Carbohydrate Research 2016, 425, 48-58. Peak annotation was performed on the chromatograms of all GOS samples based on the peak annotation made by van Leeuwen et al. for the oligosaccharide compounds. A retention window of approximately 15 seconds was applied for peak annotation.

[0190] Table 2 shows the information regarding the HPAEC-PAD peak areas of all annotated peaks, together with the information regarding the incubation conditions, sample concentration, dilution, and injection volume.

[0191]

Table 2

[0192] It was assumed that the peak area corresponded approximately to the amount of each oligosaccharide compound present in the composition. When a particular oligosaccharide compound was not identified, then "unknown" and a number were entered in the table. The identified compounds were identified either by name or by a number corresponding to the number assigned to the particular 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.

[0193] The oligosaccharide components (a) to (e) discussed above, where each X group is Glu, correspond to the following numbered entries in Table 2 above: (a) D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glu - 12 (b) D-Gal-(β1-3)-D-Gal-(β1-3)-D-Glu - 29 (c) D-Gal-(β1-3)-D-Gal-(β1-2)-D-Glu - 30 (d) D-Gal-(β1-3)-D-Gal-(β1-3)-D-Gal-(β1-4)-D-Glu - 31 (e) D-Gal-(β1-4)-D-Gal-(β1-4)-D-Glu - 11。

[0194] Prebiotic effect - Butyric acid production Butyric acid is produced by the gut microbiota that converts acetic acid and / or lactic acid (along with other substrates) into butyric acid. Since butyric acid is a secondary metabolite, it is often produced during the later stages of culture. These experiments were aimed at evaluating the differences in butyric acid production of the compositions of the present invention in parallel with a comparative GOS composition to see whether the compositions of the present invention are more effective as prebiotics. The experiments used the following procedure.

[0195] The compositions of Example 2, Comparative Example 1 and the control blank sample of the present invention were subjected to dialysis using a 0.5 kDa membrane to obtain a 5 g / l sample, which was then mixed with feces obtained from three healthy human adult subjects (Donors A, B and C). The mixture was shaken under anaerobic conditions and monitored for 48 hours for colonic fermentation products containing butyric acid (at collection points of 6, 24 and 48 hours). The distribution of oligosaccharides in the mixture was also monitored over this period using the method described above with respect to Table 2. The results show that Example 2 was well fermented by all donors, mainly during the period of 0 - 24 hours, and that butyric acid production increased at 6 hours and 48 hours compared to Comparative Example 1 and the control. The results of butyric acid analysis are shown in Figure 1. The results of oligosaccharide analysis for the samples at different time points are shown in Table 3. These results show that the oligosaccharides in the samples were actively consumed by the bacterial flora present in the fecal samples during the experiment.

[0196] To evaluate whether the treatment effect on the intestinal microbiota activity was statistically significant, three two-sided t-tests were performed between Example 2 and the control, between Comparative Example 1 and the control, and between Example 2 and Comparative Example 1 to obtain p-values. The Benjamini-Hochberg false discovery rate (FDR) was also used in this analysis. The difference between treatment effects was considered significant when the obtained p-value was less than the reference value. Table 4 below shows the differences in average butyric acid production for the compared samples over 48 hours, and the asterisks indicate whether the differences were considered significant according to the above analysis. These results indicate that the increase in butyric acid production provided by Example 2 during the period of 0 to 48 hours was statistically significant compared to the control and Comparative Example 1.

[0197]

Table 3-1

[0198]

Table 3-2

[0199]

Table 4

[0200] In summary, the present invention provides an oligosaccharide compound, for example, based on the total weight of the oligosaccharide compound present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X a ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X b ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X c wherein X a X b and X cProvided are compositions, each independently, containing a galactooligosaccharide compound selected from monosaccharides. These compositions contain relatively high amounts of oligosaccharide compounds (a), (b) and (c) and relatively high amounts of β1-3 Gal-Gal linkages as compared to known oligosaccharide compositions. These specific features of the compositions are thought to provide benefits to the health of the consumer's gut, for example, because these compositions provide an increased production of butyric acid in the consumer's gut as compared to known compositions.

[0201] Example 4 - Effect of 24-week prebiotic intervention on self-reported upper respiratory, gastrointestinal symptoms and immune markers in elite rugby union players The study evaluated the effect of 24-week B-GOS supplementation on the severity, duration and incidence of URS and GIS, salivary immunoglobulin A (sIgA), and the concentrations of C-reactive protein (CRP) and TNF-α in elite rugby union players during the competitive season.

[0202] The study was a randomized double-blind placebo-controlled trial over a 24-week period during the normal rugby union season in the Gallagher English Premiership. Forty-one healthy elite rugby union players (mean age 23.5 ± 4.7 years; body mass 103.8 ± 13.2 kg; 186.5 ± 7.8 cm) from a single club volunteered to participate in the study. All individuals were non-smokers, had no gastrointestinal medical history, and did not regularly consume probiotic-, prebiotic-, or vitamin-rich foods. Participants were matched into pairs based on body mass and playing position before random assignment to the intervention (see Table 5 below). All data were collected during the autumn and winter months in the UK (temperature range -4 to +25 °C).

[0203] During the study, a typical week for the participants included 4 to 5 days of training (lasting about 5 hours per day), one competitive match, and at least one day of rest. The training included resistance, skill, fitness, tactical, and match play exercises. Players not named to the team on the day of the competitive match had an additional day of training. The participants did not follow a diet plan made for a specific individual, but were provided with meals locally during training days. A cooked breakfast was provided before the first training session, a snack was provided after the morning session. The main meal then followed after the afternoon session. Similar meals were provided before the competitive match and a recovery drink was provided after the match. Similarly, no meal plan was provided when away from the training ground. However, individuals were instructed to avoid any foods rich in probiotics, prebiotics, and vitamins. Before each data collection visit, individuals were required to arrive after an overnight fast and avoid using mouthwash.

[0204] Supplementation Players were randomly selected to receive either 2.9 g / day of the oligosaccharide composition of the present invention (in powder form) or 2.9 g / day of a placebo (maltodextrin). Both supplements were identical in taste and color, and the supplements were blinded. The researchers, club staff, and participants remained blinded until all statistical analyses were completed. The participants were provided with the supplement pre-mixed in water at the training ground and consumed it under the observation of a club staff member. On rest days, players were instructed to mix the sachet in water and consume it at breakfast. The participants returned the used and unused sachets to evaluate supplement compliance.

[0205] Daily upper respiratory symptoms To establish the presence of URS, participants completed the Jackson questionnaire daily (Jackson et al., 1958). The presence of eight symptoms (headache, chill, sneeze, sore throat, malaise, cough, nasal discharge, nasal congestion) was evaluated on a scale of 0–3 (0 – none, 1 – mild, 2 – moderate, 3 – severe). The total symptom score for each day was summed to obtain the total Jackson symptom score. Episodes of URS were defined using the Jackson criteria applied by Martineau, Hanfia and Witt (2015), and an episode was defined as any period having a subjective impression of having a cold for at least three days, a Jackson score of 14 or more, and the presence of nasal discharge or a symptom score of less than 14 lasting for more than three days. If the symptoms of URS returned within one week, it was considered the same episode.

[0206] Weekly gastrointestinal symptoms To assess gastrointestinal symptoms (GIS), participants were required to complete a weekly gastrointestinal symptom tool (Gaskell et al. 2019). Participants were educated and advised to rate each symptom using a 10-point visual analog scale where 1–4 indicated mild GIS (i.e., there was a sensation of GIS but it was not substantially sufficient to interfere with movement), 5–9 indicated severe GIS (i.e., GIS substantially sufficient to interfere with movement), and 10 indicated very severe GIS (i.e., causing a decrease in exercise load or cessation from exercise). If no specific GIS was reported, this was rated as zero. GIS including reflux, projectile vomiting, and defecation were rated as either 0 or 10 only as their presence would result in cessation of exercise. Participants were asked to rate each symptom with reference to the previous seven days. All symptom scores were summed to obtain weekly totals and incidence rates. Additionally, the maximum possible weekly score for everyone was 190 (Gaskell et al., 2019).

[0207] Collection and analysis of sIgA At 0, 12, and 24 weeks, all participants provided saliva samples to determine sIgA. Participants rinsed their mouths with plain water and remained seated for 10 minutes before providing each sample. Unstimulated saliva samples were generated using a passive method, where participants were instructed to accumulate as much saliva as possible in their mouths for 2 minutes and then tilt their heads forward to slowly release the saliva into a collection tube for an additional 2 minutes. Samples were immediately frozen at -20°C and then at -80°C within 48 hours until analysis. For analysis, samples were completely thawed at room temperature and sIgA concentration was determined by enzyme-linked immunosorbent assay (ELISA) (Salimetrics, Philadelphia, PA). The intra-assay and inter-assay variability for sIgA were 3.2% and 7.3%, respectively, and the minimum detection level of the assay was 2.5 μg / ml, which was exceeded by all samples. sIgA flow rate was calculated by multiplying the concentration of sIgA (μg / ml) by the flow rate (ml / min), resulting in concentration measurements per unit time (μg / min) according to the manufacturer's instructions.

[0208] Collection and analysis of blood biomarkers of systemic inflammation All participants provided blood samples at 0, 12, and 24 weeks. Samples were drawn from the antecubital vein into two 10-ml vacutainers with one heparin and one EDTA coagulant (BD Vacutainer®). Plasma was used to determine the concentrations of tumor necrosis factor alpha (TNF-α) and CRP at each time point. Samples were centrifuged and immediately frozen at -20°C and then at -80°C within 48 hours until analysis. TNF-α was evaluated using a high-sensitivity ELISA, and CRP was evaluated using a normal ELISA protocol (R&D Systems). For TNF-α, the within-assay and between-assay variations were 7.5% and 5.8%, respectively, and the minimum detection level of the assay was 0.022 pg / ml, which all samples exceeded. For CRP, the within-assay and between-assay variations were 4.4% and 7.8%, respectively, and the minimum detection level of the assay was 0.022 pg / ml, which all samples exceeded.

[0209] Statistical Analysis Statistical analysis was performed using the Statistical Package for the Social Sciences (IBM SPSS version 26, Illinois, United States). All data were checked 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 evaluate differences in symptom severity. Intergroup differences in URS incidence were evaluated using the Mann-Whitney U test. Differences in URS episode duration and weeks without GIS were evaluated using an independent t-test. The secretion rates of CRP, TNF-α, and sIgA were adjusted for baseline values and evaluated using analysis of covariance (ANCOVA). Data evaluated using parametric tests are presented as mean ± standard deviation. Data presented using nonparametric tests are presented as median (range). Statistical significance was set at P<0.05.

[0210] Characteristics of the Players The final sample consisted of 33 participants (n = 17 placebo, 16 B-GOS) who successfully completed the entire 24-week supplementation period. Adherence to the supplements both during training and at home was good (B-GOS: 80.4 ± 13.9%, placebo: 78.3 ± 14%, P = 0.73). Body mass, height, age, average weekly workload, and minutes of sport played did not differ between groups (P>0.05) (see Table 5 showing the characteristics of the study participants).

[0211] [[Table 5]]

[0212] Upper respiratory symptoms (URS) The Mann-Whitney U test revealed 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 showing the summary of self-reported URS data). There was no difference in the AUC of the daily symptom scores 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 to the placebo group (9.8 ± 4.1 days) (P = 0.04).

[0213] [[Table 6]]

[0214] Gastrointestinal symptoms (GIS) Differences in GIS over the 7 days prior to day 0 were not apparent between the two groups (P = 0.53). The Mann-Whitney U test revealed that the AUC of the total weekly symptom score was lower in the B-GOS group (50 [10.5 - 139.5]) compared to the placebo group (149 [69 - 208]) (P = 0.03) (Figure 2). The upper symptom score of the AUC per week was lower in the B-GOS group compared to the placebo group (P < 0.001), but no difference was found for the lower GIS (P = 0.11) (data not shown). The number of symptom-free weeks for the total GIS and upper GIS was lower in the B-GOS group compared to the placebo group (P < 0.05), and no difference was apparent for the lower GIS (P = 0.15) (see Table 7 showing symptom-free weeks for GIS during the 24-week study).

[0215]

Table 7

[0216] Systemic inflammation No differences were observed in CRP or TNF-α at any time point between the two groups (P > 0.05).

[0217] sIgA No differences between the two study groups were observed in sIgA concentration at week 0 or week 12, but at week 24, sIgA was higher in the B-GOS group (B-GOS, median (IQR) 245 (177.9, 319.2)) compared to the placebo group (placebo median (IQR) 169 (110.6, 229.9); P = 0.006) (Figure 3). No differences were observed in sIgA secretion rate at week 0 or week 12, but differences were 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).

[0218] Conclusions of the prebiotic intervention The main findings of this study were that daily supplementation with B-GOS decreased the duration of URS and the incidence of GI symptoms in elite rugby union players over a 24-week period. Furthermore, B-GOS increased the concentration and secretion rate of sIgA at 24 weeks when compared to the placebo group. These findings indicate that B-GOS can modulate the immune and GI systems to suppress URS and GI discomfort associated with elite rugby union play.

[0219] Illness can have an adverse impact on the training potential and match readiness of athletes (Cunniffe et al., 2009; Tiernan et al., 2020; Keaney et al., 2021). To the inventors' knowledge, this is the first study to evaluate the effect of prebiotic dietary interventions on URS and GIS in the athlete population. The 22% reduction in the duration of URS episodes is clinically important and relevant for athletes and coaches alike, ensuring that athletes can return to play sooner after URS episodes.

[0220] B-GOS has previously been shown to promote the growth of Bifidobacterium in the human gut, conferring numerous health benefits such as reduced systemic inflammation and improved immune responses in elderly and overweight populations (Vulevic et al., 2008; Vulevic et al., 2013). Furthermore, increases in fecal short-chain fatty acid (SCFA) production, as well as potential enhancements in epithelial integrity and mucosal immunity, have also been reported (Hernot et al., 2009; Mariadason, Barkla & Gibson, 1997). Indeed, in the present study, B-GOS increased sIgA concentration and secretion rate at 24 weeks when immunity may have been compromised due to increased training load, as seen by the decrease in the placebo group. This suggests that active manipulation of the gut microbiota can support mucosal immunity and sIgA production.

[0221] B-GOS is thought to reduce systemic inflammation through increased growth of Bifidobacterium. Bifidobacterium can amplify the sampling of dendritic cells in the gut, alter the differentiation of naive T cells and increase T cells expressing forkhead box protein P3 (FoxP3), thereby producing an anti-inflammatory effect (McLoughlin & Mills, 2011).

[0222] It should be noted that the total number of URS episodes in this study was less than previously reported for winter. Elite rugby union players have been reported to experience four URS episodes per season (Cunniffe et al., 2009), with the highest incidence during winter. We reported significantly lower rates 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 still improve the URS period even when the incidence is low. We also found that participants continued to train despite showing URS. This finding has also been found elsewhere in similar cohorts and may have been because participants were required to train while at the training site (Cunniffe et al., 2009). They may also fear the possibility of being dropped from the next game. This is a limitation of collecting self-reported data, and future studies should use molecular tests to determine infection. Another limitation was the frequency of sIgA and cytokine measurements. B-GOS may have reduced the URS period through enhancement of sIgA.

[0223] In conclusion, 24 weeks of prebiotic B-GOS supplementation decreased the URS period, the incidence and severity of GIS, and increased the sIgA secretion rate in elite rugby union players. These findings indicate that B-GOS has the potential to regulate immune function, reduce illness, and improve the ability of athletes to train and compete.

[0224] Although several preferred embodiments have been shown and described, it will be recognized by those skilled in the art that various changes and modifications can be made without departing from the scope of the invention, as defined in the appended claims.

[0225] Throughout this specification, the term "comprising" or "comprises" means including the stated element(s) (singular or plural), but not precluding the presence of other elements. The term "consisting essentially of" or "consists essentially of" means including the stated element, but excluding other elements except for substances present as impurities, inevitable substances resulting from the processes used to provide the element, and other elements added for purposes other than achieving the technical effect of the invention. Typically, when referring to a composition, a composition consisting essentially of a set of components contains less than 5% by weight, typically less than 3% by weight, more typically less than 1% by weight of unspecified components.

[0226] The term "consisting of" or "consists of" means including the stated element, but precluding the addition of other elements.

[0227] Whenever appropriate and depending on the context, the use of the term "comprises" or "comprising" can also be interpreted as encompassing or including the meaning "consists essentially of" or "consisting essentially of", and also as including the meaning "consists of" or "consisting of".

[0228] To avoid misunderstanding, when the amount of a component in a composition is stated in weight %, this means the weight percentage of the specific component in relation to the total composition being referred to. For example, "here, the oligosaccharide compound contains up to 35% by weight of disaccharide" means that 35% by weight of the oligosaccharide compound in the composition is provided by the disaccharide.

[0229] Any feature described herein may be used individually or in combination with each other as required, and in particular in the combinations described in the appended claims. Any feature for each aspect or exemplary embodiment of the invention described herein should also be read as being applicable, as required, to any other aspect or exemplary embodiment of the invention. In other words, a person skilled in the art reading this specification should consider any feature for each exemplary embodiment of the invention to be interchangeable and combinable between different exemplary embodiments.

[0230] Attention is directed to all papers and documents that have been filed in connection with this application simultaneously with or prior to this specification and that have been published for public inspection with this specification, and the contents of all such papers and documents are hereby incorporated herein by reference.

[0231] All of the features disclosed in this specification (including any appended claims and drawings), and / or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations in which at least some of such features and / or steps are mutually exclusive.

[0232] Each feature disclosed in this specification (including any appended claims and drawings) may be replaced by alternative features that serve the same, equivalent or similar purpose, unless expressly stated otherwise. That is, each feature disclosed is only an example of a general series of equivalent or similar features, unless expressly stated otherwise.

[0233] The present invention is not limited to the details of the foregoing embodiment(s). The present invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any appended claims and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

Claims

1. A composition comprising an oligosaccharide compound for use in the prevention, improvement, or treatment of conditions and / or diseases of the upper respiratory system (URS) and gastrointestinal tract (GI), wherein the oligosaccharide compound is: Based on the total weight of the oligosaccharide compounds present in the composition, (a) at least 8% by weight of Gal-(β1-3)-Gal-(β1-4)-X ａ ; (b) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-X ｂ ; and (c) at least 5% by weight of Gal-(β1-3)-Gal-(β1-2)-X ｃ Includes, in the formula, X ａ , X ｂ and X ｃ Each of these is independently selected from monosaccharides, forming a composition.

2. Oligosaccharide compounds: Based on the total weight of oligosaccharide compounds present in the composition, (d) at least 3% by weight of Gal-(β1-3)-Gal-(β1-3)-Gal-(β1-4)-X ｄ Includes, in the formula, X ｄ The composition for use according to claim 1, which is selected from monosaccharides.

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

4. A composition for use according to claim 1 or claim 2, wherein the weight percentage ratio of compound (a) to compound (b) is 1:1 to 3:

1.

5. A composition for use according to claim 1 or claim 2, wherein the weight percentage ratio of compound (a) to compound (c) is 1:1 to 3:

1.

6. A composition for use according to claim 1 or claim 2, wherein the weight percentage ratio of compound (b) to compound (c) is 2:1 to 1:

2.

7. Oligosaccharide compounds: Based on the total weight of oligosaccharide compounds present in the composition, (e) at least 5% by weight of Gal-(β1-4)-Gal-(β1-4)-X ｅ Includes, in the formula, X ｅ a composition for use according to claim 1 or claim 2, selected from monosaccharides.

8. X ａ , X ｂ , X ｃ , X ｄ and X ｅ A composition for use according to claim 1 or 2, wherein each of these is independently selected from glucose, fucose, arabinose, xylose, rhamnose, mannose, galactose, ribose, lyxose, allose, altrose, growth, idose, talose, psicose, fructose, sorbose, tagatose, galactosamine, glucosamine, and N-acetylglucosamine or a mixture thereof.

9. X ａ , X ｂ , X ｃ , X ｄ and X ｅ A composition for use according to claim 1 or claim 2, wherein each of the elements is Glc.

10. X ａ , X ｂ , X ｃ , X ｄ and X ｅ A composition for use according to claim 1 or claim 2, wherein each of the elements comprises Fuc.

11. A composition for use according to claim 1 or 2, wherein 40 to 55% of the Gal-Gal bonds in the oligosaccharide compound are 1-3 bonds.

12. A composition for use according to claim 1 or 2, comprising at least 50% by weight of an oligosaccharide compound, preferably in the form of a syrup.

13. A composition for use according to claim 1 or 2, comprising at least 75% by weight of an oligosaccharide compound, preferably in the form of a powder.

14. A composition for use according to claim 1 or claim 2, wherein the oligosaccharide compound comprises at least 25% by weight of a trisaccharide.

15. A composition for use according to claim 1 or claim 2, wherein the oligosaccharide compound comprises at least 10% by weight of a tetrasaccharide.

16. A composition for use according to claim 1 or claim 2, comprising 80% by weight of trisaccharides, tetrasaccharides and higher oligosaccharides.

17. A composition for use according to claim 1 or claim 2, comprising 80% by weight of trisaccharides.

18. A composition for use according to claim 1 or 2, wherein the composition is in the form of a nutritional supplement.

19. A composition for use according to claim 1 or 2, wherein the composition is in the form of a pharmaceutical or nutritional supplement.

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

21. The composition for use according to claim 1 or claim 2, wherein the upper GI condition and / or disease is selected from one or more of the following upper GI conditions and diseases: indigestion, stomach upset, difficulty swallowing, heartburn, acid reflux, reflux esophagitis, gastroesophageal reflux disease, gastritis, small intestinal bacterial overgrowth, and dyspepsia.

22. The composition for use according to claim 1 or 2, wherein the condition and / or disease of the upper URS is an upper respiratory tract infection (URTI).

23. The composition for use according to claim 1 or 2, wherein the URS and GI condition and / or disease is in an individual who is an athlete and / or is undergoing or intending to undergo physical training.

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