Oligosaccharides for glycemic control

Abstract

Methods for treating glucose-related metabolic disorders in a subject are provided. In some aspects, the methods comprise enterally administering an effective amount of a beta-glucan oligosaccharide prior to and / or during intake of a glucose source in the subject.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 383,464, filed on November 11, 2022, the entire contents of which are hereby incorporated by reference.

[0003] The entire contents of each of the following applications are incorporated into this application by reference for all purposes to the extent not inconsistent with the present disclosure: PCT patent application No. PCT / US22 / 29065 filed on May 12, 2022 (WO 2022 / 241163 A1), PCT patent application No. PCT / US2018 / 038350 filed on June 19, 2018 (WO 2018 / 236917 A1), PCT patent application No. PCT / US2020 / 035748 filed on June 2, 2020 (WO 2020 / 247389 A1), PCT patent application No. PCT / US2020 / 060297 filed on November 12, 2020 (WO 2021 / 097138 A1). Technical Field

[0004] The disclosure herein generally relates to oligosaccharides, compositions comprising oligosaccharides, methods of obtaining oligosaccharides and oligosaccharide compositions, methods of using oligosaccharides or oligosaccharide compositions to modulate microbiota and their metabolites, and methods of using oligosaccharides or oligosaccharides as therapeutic agents for health applications, including for managing glucose levels in the blood. Background Art

[0005] A chronic medical condition or disease is a condition that persists over an extended period of time, requires ongoing medical attention, limits activities of daily living, or both. In some cases, symptoms may flare up and relapse, while in other cases, they persist. Chronic disease is extremely common and a major contributor to impaired quality of life and the economic burden of health. For example, chronic diseases such as heart disease, cancer, and diabetes are the leading causes of death and disability in the United States and are the primary driver of the country's $3.8 trillion annual healthcare costs.

[0006] Hyperglycemia, or high blood sugar, can cause serious damage to many body systems over time, especially the nerves and blood vessels. Long-term complications of hyperglycemia can include heart disease, stroke, diabetic retinopathy, kidney failure, and poor blood flow to the extremities. Lowering the glycemic response has been associated with a reduced risk of diabetes, a condition characterized by chronic hyperglycemia. Post-meal blood sugar levels are related to the amount of insulin the body produces and the amount of glucose absorbed by the small intestine, and are influenced by a variety of environmental factors, such as diet. The Western diet, which contains a large amount of high-sugar and high-starch foods, as well as changes in the gut microbiome caused by a low-fiber diet, can affect multiple mechanisms in our bodies related to glucose absorption. Most current treatments are aimed at treating the consequences of hyperglycemia, rather than the cause of the impaired metabolism. This approach is not very effective, so there is still a need to find safe ways to address the underlying causes or at least manage the impaired metabolism over a longer period of time with few or no adverse side effects.

[0007] One strategy for controlling blood glucose levels is to target mechanisms that influence glucose absorption at the interface between the intestinal wall and the intestinal lumen. Controlling intestinal glucose absorption in the intestinal brush border membrane could help reduce the risk of hyperglycemia. The glucose transporter sodium-glucose transporter 1 (SGLT1) is responsible for the active transport of glucose across the brush border membrane of the small intestine, and inhibition of SGLT-1 can significantly reduce intestinal glucose absorption in patients at risk for diabetes. Several polysaccharides have been reported to significantly reduce SGLT-1 expression not only in the intestinal mucosa of diabetic mice but also in Caco-2 cells. Furthermore, pure β-glucan from baker's yeast (referred to as linear β-(1→3)-glucan) has been reported to significantly improve blood glucose regulation in normal mice and reduce glucose transport across Caco-2 monolayers (Cao et al. 2016).

[0008] Another strategy to reduce the intake of carbohydrates from food and their entry into the bloodstream is to target α-glucosidases in the brush border membrane of the small intestine. Dietary starch is broken down into small carbohydrate chains by α-amylase in the stomach, which are then broken down into monosaccharides in the small intestine by an enzyme complex called α-glucosidase. α-glucosidase can be competitively inhibited by compounds that resemble short starch-derived carbohydrates. Consequently, monosaccharide formation is reduced, and less glucose is absorbed by the host, leading to a reduction in the food-induced postprandial blood glucose rise (Benalla, W et al. 2010). The antidiabetic drug acarbose (a pseudotetrasaccharide, O-4,6-dideoxy-4-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)-2-cyclohexen-1-yl]amino]-α-d-glucopyranosyl-(1→4)-O-α-d-glucopyranosyl(1→4)-d-glucose) is an α-glucosidase inhibitor. However, acarbose has been reported to cause gastrointestinal side effects (particularly flatulence and diarrhea) because it also inhibits α-amylase activity.

[0009] Furthermore, certain oligosaccharides that are not digested by the host can reach the lower intestine, where they are fermented by the intestinal microbiota. Fermentation of these oligosaccharides by specific bacterial microorganisms produces short-chain fatty acids, which activate GPR41 and GPR43 receptors. Activation of these receptors stimulates the secretion of glucagon-like peptide-1 (GLP-1). This gut hormone slows gastric emptying, reduces glucagon secretion, and regulates insulin secretion (Everard and Cani, 2014).

[0010] Optimal glycemic control is the cornerstone of managing prediabetes and diabetes. Both fasting and postprandial glucose levels are associated with the risk of complications and influence the measurement of glycated hemoglobin (A1C). A1C levels >7.0% are associated with a significantly increased risk of microvascular and cardiovascular (CV) complications. Postprandial glucose is an important component of overall hyperglycemia and may be the major component in patients approaching the A1C extremes and in the elderly. For example, postprandial glucose has been shown to be the major contributor to total glycemic excursions in patients with T2D (i.e., well-controlled diabetes or prediabetes) who have HbA1c <8%. Prediabetes is increasing rapidly worldwide and is primarily related to age and BMI. Therefore, controlling postprandial glucose in overweight and obese individuals may play a role in preventing the progression of prediabetes to diabetes.

[0011] Current treatments for reducing postprandial glucose response typically include α-glucosidase inhibitors such as acarbose, or SGLT-1 inhibitors such as empagliflozin, canagliflozin, and dapagliflozin. However, these drugs have side effects. Other approaches include using nutritional formulas containing complex carbohydrates as meal replacements for foods containing simple carbohydrates. However, the need to consume standard meal replacements typically leads to taste fatigue and loss of compliance over time.

[0012] Therefore, there remains a need for safe and effective interventions that allow subjects to manage glucose responses long-term, particularly postprandial glucose responses. Summary of the Invention

[0013] Provided are methods for adjusting the microbial community and related bioactive metabolites in vitro and / or in the gastrointestinal tract of a subject by administering one or more oligosaccharide compositions of different structures. Depending on the oligosaccharide structure, the microbial community structure (e.g., microbial abundance level and composition) and metabolism respond in different ways. Also provided are methods for treating diseases, conditions, disorders, and / or indications related to gastrointestinal health and / or metabolic disorders. Also provided are novel oligosaccharide compositions and their structural features. Typically, the oligosaccharide compositions are, but not necessarily, derived from natural products.

[0014] Also provided are improved methods for producing polysaccharide cleavage products and / or oligosaccharide mixtures by reacting polysaccharides in a reaction mixture with a Fenton reagent having a peroxide and a metal ion, and lysing the treated polysaccharide with an alkali to produce the polysaccharide cleavage products and / or oligosaccharide mixtures. In one embodiment, the improvement includes using copper metal ions in combination with hydrogen peroxide to increase the reaction efficiency and the yield of the cleavage products and oligosaccharide mixtures.

[0015] In one aspect, the present disclosure provides a method for treating a glucose-related metabolic disorder in a subject, the method comprising administering an effective amount of β-glucan oligosaccharide to the subject enterally before and / or during the subject's intake of a glucose source. In a more specific aspect, the glucose-related metabolic disorder is prediabetes, diabetes, including type 1 diabetes, type 2 diabetes, gestational diabetes, or metabolic syndrome.

[0016] In another aspect, the present disclosure provides a method for reducing a postprandial glucose response in a subject, the method comprising enterally administering to the subject an effective amount of β-glucan oligosaccharide prior to and / or during ingestion of a glucose source by the subject. In a more specific aspect, the subject being treated is at risk for diabetes, is overweight and / or obese, is pregnant, and / or has prediabetes or diabetes.

[0017] In another aspect, the present disclosure provides a method for reducing the risk of a prediabetic and / or obese subject developing type 2 diabetes, the method comprising attenuating the subject's postprandial glucose response by enterally administering to the subject an effective amount of β-glucan oligosaccharide before and / or during ingestion of a glucose source by the subject.

[0018] In another aspect, the present disclosure provides a method for reducing HbA1c levels in a subject, the method comprising attenuating the subject's postprandial glucose response for at least 2 months by enterally administering to the subject an effective amount of β-glucan oligosaccharide before and / or during ingestion of a glucose source by the subject.

[0019] In embodiments of the above aspects of the present disclosure, the subject is at risk for diabetes, is overweight and / or obese, is pregnant, and / or has prediabetes or diabetes. In further embodiments, about 0.5 g to about 20 g of beta-glucan oligosaccharide may be administered to the subject. For example, about 0.75 g to about 15 g or about 0.75 g to about 7.5 g of beta-glucan oligosaccharide may be administered to the subject. In further embodiments, the beta-glucan oligosaccharide may be administered to the subject less than about 2 hours, 1 hour, or 30 minutes before the subject ingests a glucose source, for example, less than about 15 minutes before the subject ingests a glucose source. In further embodiments, the beta-glucan oligosaccharide is co-administered with the glucose source. In some embodiments, the beta-glucan oligosaccharide is administered simultaneously or almost simultaneously with the glucose source. In some embodiments, the beta-glucan oligosaccharide is administered before the glucose source or simultaneously with the glucose source. In some embodiments, the β-oligosaccharide and the glucose source are administered as part of the same treatment, but at different times, e.g., 8 hours apart, 12 hours apart, 1 day apart, 2 days apart, or 1 week apart. In further embodiments, the β-glucan oligosaccharide inhibits salivary amylase and / or pancreatic amylase. In further embodiments, the β-glucan oligosaccharide inhibits SGLT1 glucose transporter and / or inhibits α-glucosidase. In further embodiments, the β-glucan oligosaccharide comprises β-1,3 or β-1,4 linked glucose residues. In further embodiments, the β-glucan oligosaccharide comprises both β-1,3 and β-1,4 linked glucose residues. In further embodiments, the β-glucan oligosaccharide comprises a ratio of β-1,3 linked glucose residues to β-1,4 linked glucose residues of 1:1 to 1:5, e.g., 1:1 or 1:2 or 1:3 or 1:4 or 1:5. In a further embodiment, the β-glucan oligosaccharide has a weight average molecular weight of less than 10,000 Da, or less than 8,000 Da, or less than 7,500 Da, or less than 5,000 Da, optionally greater than 500 Da, or greater than 1,000 Da, or greater than 2,000 Da. In a further embodiment, the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue. In a further embodiment, the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue, and each oligosaccharide comprises both β-1,3 glucose residues and β-1,4 glucose residues. In further embodiments, the β-glucan oligosaccharide comprises 3 to 30, or 3 to 25, or 5 to 30, or 5 to 25 subunits, or any subrange thereof, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue.In further embodiments, the β-glucan oligosaccharide comprises 3 to 30, or 3 to 25, or 5 to 30, or 5 to 25 subunits, or any subrange thereof, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue, and each oligosaccharide comprises both β-1,3 glucose residues or β-1,4 glucose residues. In further embodiments, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C. In further embodiments, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C, or any subranges thereof.

[0020] In yet another aspect, the present invention provides a synthetic composition for treating a glucose-related metabolic disorder, reducing a subject's postprandial glucose response, reducing the risk of progression to type 2 diabetes in a prediabetic subject and / or an obese subject, and / or for reducing a subject's HbA1c level, the synthetic composition comprising, consisting essentially of, or consisting of an effective amount of β-glucan oligosaccharides. In a further embodiment, the β-glucan oligosaccharide comprises β-1,3 and β-1,4 linked glucose residues. In a further embodiment, the β-glucan oligosaccharide comprises both β-1,3 and β-1,4 linked glucose residues. In a further embodiment, the β-glucan oligosaccharide comprises a ratio of β-1,3 linked glucose residues to β-1,4 linked glucose residues of 1:1 to 1:5, e.g., 1:1 or 1:2 or 1:3 or 1:4 or 1:5 or any subrange thereof. In a further embodiment, the β-glucan oligosaccharide mixture has a weight average molecular weight of less than 10,000 Da, or less than 8,000 Da, or less than 7,500 Da, or less than 5,000 Da, or any subrange thereof. In a further embodiment, the β-glucan oligosaccharide comprises from about 3 to about 50 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue. In a further embodiment, the β-glucan oligosaccharide comprises from about 3 to about 50 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue, and each oligosaccharide comprises both a β-1,3 glucose residue and a β-1,4 glucose residue. In a further embodiment, the β-glucan oligosaccharide comprises from 3 to 30, or from 3 to 25, or from 5 to 30, or from 5 to 25 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue. In further embodiments, the β-glucan oligosaccharide comprises 3 to 30, or 3 to 25, or 5 to 30, or 5 to 25 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue, and each oligosaccharide comprises both β-1,3 glucose residues and β-1,4 glucose residues. In further embodiments, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C. In further embodiments, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C, or a dynamic viscosity of about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C.

[0021] In yet another aspect, the present invention provides a pharmaceutical pack for treating a glucose-related metabolic disorder, or for reducing a postprandial glucose response in a subject, or for reducing the risk of a prediabetic and / or obese subject developing type 2 diabetes, and / or for reducing HbA1c levels in a subject, the pharmaceutical pack comprising, consisting essentially of, or consisting of at least 14 individual doses of an effective amount of β-glucan oligosaccharide, wherein the β-glucan oligosaccharide is as described herein above.

[0022] In another aspect, the present disclosure provides use of the β-glucan oligosaccharide as described herein above for treating a glucose-related metabolic disorder, or for attenuating a subject's postprandial glucose response, or for reducing the risk of a prediabetic subject and / or an obese subject developing type 2 diabetes, and / or for reducing a subject's HbA1c level.

[0023] In another aspect, the present disclosure provides use of a β-glucan oligosaccharide as described herein above for the preparation of a medicament for treating a glucose-related metabolic disorder, or for attenuating a subject's postprandial glucose response, or for reducing the risk of a prediabetic subject and / or an obese subject developing type 2 diabetes, and / or for reducing a subject's HbA1c level.

[0024] In an embodiment, glucose-related metabolic disorders herein include, but are not limited to, diabetes (type 1 diabetes, type 2 diabetes, and gestational diabetes), as well as prediabetes and metabolic syndrome.

[0025] Additional aspects and embodiments of the present disclosure will become apparent to those of ordinary skill in the art upon reading the following detailed description, non-limiting examples, and accompanying figures.

[0026] Without wishing to be bound by any particular theory, this paper may discuss ideas or understandings of the underlying principles associated with the devices and methods disclosed herein. It should be recognized that regardless of the ultimate correctness of any mechanistic explanation or hypothesis, embodiments or aspects of the present invention may still be operational and useful. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 : Relative transporter-specific accumulation of AMG (%).

[0028] Figure 2 : Inhibitory effect of CLX115 on α-glucosidase.

[0029] Figure 3 : Inhibitory effects of different β-glucan sources on α-glucosidase.

[0030] Figure 4A-4B : Butyrate production levels of CLX115 in vitro (SHIME model). Figure 4A Proximal colon findings are depicted. Figure 4B Distal colon findings are depicted.

[0031] Figure 5A-5B : Propionate production levels of CLX115 in vitro (SHIME model). Figure 5A Proximal colon findings are depicted. Figure 5B Distal colon findings are depicted.

[0032] Figure 6 : Gas generation level (kPa).

[0033] Figures 7A-7D : RID chromatogram. Figure 7A Results for CLX115-PS are depicted. Figure 7B Results for CLX115Cu are depicted. Figure 7C Results for CLX115 are depicted. Figure 7D Results for CLX112 are depicted.

[0034] Figure 8A-8B : Evaluation of the inhibitory effect of CLX115 on α-glucosidase. Figure 8A The change in maltose concentration level over time is depicted. Figure 8B The changes in glucose concentration over time are depicted.

[0035] Figure 9A-9B : In vivo assessment of blood glucose levels. Figure 9A and Figure 9B In the figures, "ctrl" refers to the control, in which only maltose was administered. Figure 9A Depicted are blood glucose levels measured at different time points after carbohydrate gavage in two groups of mice (n=15 / group). Figure 9B Peak blood glucose levels 30 minutes after carbohydrate gavage are depicted.

[0036] Statement Regarding Incorporation by Reference and Modifications

[0037] Generally, the terms and phrases used herein have art-recognized meanings, which can be found by reference to standard texts, journal references and context known to those skilled in the art. The following definitions are provided to clarify their specific use in the context of the present invention.

[0038] The following abbreviations are used herein: Deoxyhex refers to deoxyhexose; DP refers to degree of polymerization; Glc refers to glucose; Hex refers to hexose; HexA refers to hexadecanoate; Man refers to mannose; Pent refers to pentose; and PS refers to polysaccharide.

[0039] As used herein, when the terms "about" and "approximately" are used to modify an amount specified by a numerical value or a numerical range, the terms indicate that slight deviations from the stated value can be used to achieve substantially the same result as the stated value. In cases where this definition is not applicable or extremely difficult to apply, the terms "about" and "approximately" indicate reasonable deviations from the value known to those skilled in the art, for example, if "X" is the value, for example, "about X" or "approximately X" would indicate a value of 0.9X to 1.1X, for example, a value of 0.95X to 1.05X, or a value of 0.98X to 1.02X, or a value of 0.99X to 1.01X. Any reference to "about X" or "approximately X" where X is a value disclosed herein specifically refers to at least the following values: X, 0.9X, 0.91X, 0.92X, 0.94X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, 1.05X, 1.06X, 1.07X, 1.08X, 1.09X, and 1.1X, as well as values within this range.

[0040] As used herein, when referring to one or more oligosaccharides (or any similar context), the term "collectively comprise" or "collectively comprises" or similar terms means that the one or more oligosaccharides as a whole have the indicated composition or property. For example, a given composition may contain two different oligosaccharides (e.g., a first oligosaccharide comprises a glucose subunit but does not contain an arabinose subunit, and a second oligosaccharide comprises an arabinose subunit but does not contain a glucose subunit). In this case, the two different oligosaccharides (i.e., "the one or more oligosaccharides") collectively comprise a glucose subunit and an arabinose subunit. Similarly, the term "collectively" has the same meaning in any similar context (e.g., when referencing a composition, etc.) to indicate that any components present together have the indicated characteristics or properties (e.g., NMR analysis or type or number of connections, etc.).

[0041] Unless otherwise stated, any viscosity measurements or properties reported herein use water as the solvent.

[0042] In some aspects, the compositions or compounds disclosed herein, such as oligosaccharides or oligosaccharide compositions, are isolated or substantially purified. In one embodiment, the isolated or purified oligosaccharides or oligosaccharide compositions are at least partially isolated or substantially purified, as will be understood in the art. In some aspects, the substantially purified compositions, oligosaccharides, or preparations disclosed herein have a chemical purity of 95%, optionally 99% for some applications, optionally 99.9% for some applications, optionally 99.99% for some applications, and optionally 99.999% for some applications.

[0043] As used herein, the term "polysaccharide" refers to a polysaccharide or a material comprising a polysaccharide, in either case, wherein at least the polysaccharide component can be cleaved by the COG method disclosed herein. In addition, as used herein, the term "polysaccharide" refers to any carbohydrate polymer (to which other non-carbohydrate moieties may also be attached, e.g., glycoproteins, proteoglycans, glycopeptides, glycolipids, glycoconjugates, glycosides, or any combination thereof). In addition, as used herein, "polysaccharide" refers to a polymer of monosaccharide units having more than 30 monosaccharide units, and its length can reach hundreds of thousands of monosaccharides. A polysaccharide can be a linear polymer, a branched polymer, a main linear polymer with pendant sugar monomers, or any combination thereof.

[0044] As used herein, the term "peroxidase" refers to a compound containing an oxygen-oxygen bond that can naturally produce RO and / or ROO species under the action of light, temperature, or a catalyst (e.g., metals and enzymes), where "R" refers to a hydrogen or carbon group attached to the rest of the molecule. In one aspect, the peroxidase is hydrogen peroxide.

[0045] The "degree of polymerization" or "DP" of an oligosaccharide refers to the total number of sugar monomer units (also referred to herein as subunits) that are part of a specific carbohydrate. For example, a tetragalacto-oligosaccharide has a DP of 4, with 3 galactose moieties and 1 glucose moiety. When used to describe a group of oligosaccharides (e.g., an oligosaccharide composition), DP generally refers to the average DP of the oligosaccharides in the composition. In some aspects, the DP of an oligosaccharide is referred to as "DP#," where "#" corresponds to an integer representing the total number of sugar monomer units (or the average value if used to describe a group of oligosaccharides) (e.g., "DP3" represents a degree of polymerization of 3). For the oligosaccharides or oligosaccharide compositions discussed herein having a DP in the range of 3-9, the DP includes a deviation of up to ±2 monomer units of the value. For oligosaccharides or oligosaccharide compositions having a DP of 10 or greater, the DP includes a deviation of up to ±20% of the value, for example, an oligosaccharide having a DP of 20 may have a DP of 16, 17, 18, 19, 20, 21, 22, 23, or 24.

[0046] As used herein, when the amount of a component is expressed in weight or mole percentage, unless otherwise indicated, it refers to the amount on a dry basis. As used herein, "dry basis" means without water or other solvents. For example, when a composition contains 10 g glucose, 40 g xylose, and 50 g water, this means that the composition contains 25% (mass % or weight %) glucose on a dry basis, but the glucose is present in the composition at a concentration of 10% (mass % or weight %).

[0047] As used herein, "live biotherapeutic agent" refers to a therapeutic product or pharmaceutical product that contains a living microorganism (e.g., archaea, bacteria, algae, fungi (e.g., yeast), or any combination thereof) as an active ingredient. In some aspects, a live biotherapeutic agent can include an engineered (e.g., genetically modified) or non-engineered living organism, or a mixture of engineered and non-engineered living organisms.

[0048] "Prebiotics" or "prebiotic nutrients" are generally non-digestible or partially digestible (i.e., digestible by a subject / human / animal, and not digestible by microorganisms) food ingredients that, when ingested, have a beneficial effect on the host by selectively stimulating the growth and / or activity of one or a limited number of microorganisms in the gastrointestinal tract, urogenital system, or other parts of the host. As used herein, the term "prebiotic" refers to such non-digestible or partially digestible food ingredients in a state that is not naturally present (e.g., after purification, chemical synthesis, or enzymatic synthesis, other than, for example, in whole human milk).

[0049] "Probiotic" refers to live microorganisms that, when administered in adequate amounts, confer a health benefit on the host.

[0050] As used herein, "stripping reaction" or "stripping" as applied to the disclosed methods refers to the sequential alkaline degradation of carbohydrates by a mechanism that releases monomer units from the reducing end of the polymer.

[0051] As used herein, preferably, " cleavage agent " or " cleavage reagent " applied to the disclosed method refers to a single or collective non-Arrhenius base and / or weak Arrhenius base for cleaving polysaccharides after peroxyhydroxyl oxidation of the polysaccharide. In some aspects, the cleavage agent or cleavage reagent destroys the glycosidic bond in the polysaccharide, and the glycosidic bond can be present between any two sugars of the polysaccharide. In an embodiment, the cleavage reagent (cleavage initiator) can also be and preferably is a peroxide quenching reagent, which can be used in combination with another compatible peroxide quencher in either case, and the other compatible peroxide quencher can be a cleavage agent or not. In some aspects, the cleavage reagent can be an enzyme. In some aspects, the cleavage reagent enzyme can be a glycoside hydrolase, a cleavage polysaccharide monooxygenase, a glycosyltransferase, a transglycosidase, a polysaccharide lyase, a carbohydrate binding assembly, a glycosyltransferase, a carbohydrate esterase, a mixture comprising two or more of the above enzymes, or any enzyme with carbohydrate activity. In some aspects, the cleavage reagent can be a solid phase acid catalyst or a solid phase base catalyst.

[0052] As used herein, "base" refers to a compound or collection of compounds that can accept hydrogen ions from carbohydrates, water, or non-aqueous solvents oxidized by hydrogen peroxide. The term "base" can include Lewis bases, non-Arrhenius bases, weak Arrhenius bases, other molecules that produce hydroxide ions by decomposition, Lewis bases, non-Arrhenius bases, or weak Arrhenius bases, or other compounds that can accept hydrogen ions from carbohydrates oxidized by hydrogen peroxide. As used herein, unless otherwise indicated, "base" explicitly does not refer to strong Arrhenius bases (e.g., Na + OH - , K + OH - or Ca +2 (OH - )2).

[0053] As used herein, "ammonium bicarbonate" as applied to the disclosed methods refers to solid ammonium bicarbonate, and / or an aqueous solution comprising: ammonium and bicarbonate; ammonium, OH - and CO2; ammonia, H2O and CO2; or any of the foregoing and the balance thereof.

[0054] As used herein, "ammonium hydroxide" suitable for use in the disclosed methods refers to: aqueous ammonium hydroxide solutions, and / or solutions comprising: ammonia and H2O; ammonium and OH - ; ammonia and OH - ; or any of the foregoing products and their equilibrium products.

[0055] As used herein, a "strong Arrhenius base" as applied to the disclosed methods refers to a compound that completely dissociates in water to release one or more hydroxide ions into solution. As used herein, a "strong Arrhenius base" as applied to the disclosed methods specifically refers to KOH, NaOH, Ba(OH)2, CsOH, Sr(OH)2, Ca(OH)2, LiOH, and RbOH.

[0056] As used herein, a "weak Arrhenius base" as applied to the disclosed methods refers to a compound that does not completely dissociate in water to release one or more hydroxide ions into solution, such as ammonium hydroxide, H2O, etc. As "weak Arrhenius base" is used herein, there is no compound that meets the definition of both a strong Arrhenius base and a weak Arrhenius base.

[0057] As used herein, "non-Arrhenius base" as applied to the disclosed methods refers to a compound or atom that can donate electrons (e.g., a Lewis base), accept protons (e.g., a Bronsted-Lowry base), or release hydroxide ions (NH4HCO3) upon decomposition thereof, but does not specifically qualify as an Arrhenius base.

[0058] As used herein, a "Lewis base" as applied to the disclosed methods refers to a compound or atom (e.g., F) that can donate an electron pair. - Benzene, H - , pyridine, acetonitrile, acetone, urea, etc.).

[0059] As used herein, a "Bronsted-Lowry base" employed in the disclosed methods refers to a compound or atom (eg, methanol, formaldehyde, ammonia, etc.) that can accept or combine with a hydrogen ion.

[0060] As used herein, a "peroxide quenching agent" as used in the disclosed methods refers to a compound or atom (e.g., ammonium hydroxide, ammonium bicarbonate, ammonia, etc.) that is not a strong Arrhenius base and can convert hydrogen peroxide, peroxyl radicals, and peroxyhydroxyl radicals into a less reactive or non-reactive state. In some aspects, the peroxide quenching agent as defined herein converts hydrogen peroxide and the free radicals generated by hydrogen peroxide into a less reactive species (e.g., water). In some aspects, the peroxide quenching agent can reduce the concentration of hydrogen peroxide to zero, less than 5 mg / L, less than 10 mg / L, less than 25 mg / L, or less than 50 mg / L. In some aspects, the peroxide quenching agent can form water, hydroxide ions, or oxygen. In some aspects, enzymes can be used to quench the peroxide species. In some aspects, these enzymes can include catalase. In some aspects, these enzymes can be derived from animal sources. In some aspects, these enzymes can be derived from bovine liver. In some aspects, the enzymes can be derived from microbial sources. In some aspects, the enzymes can be recombinant. In certain aspects, different enzymes can be mixed to quench peroxide species.

[0061] As used herein, "nitrogen-based" as applied to the disclosed methods refers to a compound comprising at least one nitrogen atom and four substituent groups (which may comprise any combination of lone electron pairs, hydrogen, or carbon atoms) (e.g., ammonia, sodium amide, trimethylamine, diethylamine, N,N-diisopropylethylamine, urea, pyridine, ammonium hydroxide, ammonium bicarbonate, etc.). Exemplary nitrogen-based, peroxide quenching, and PS cleavage agents are listed in Table 1 below. Nitrogen-based reagents may have unsubstituted or substituted ammonium groups and may exist in neutral and / or ionic form.

[0062] Table 1. Exemplary polysaccharide (PS) cleaving agents and / or peroxide quenchers.

[0063]

[0064]

[0065] As used herein, a "reaction mixture" refers to a mixture comprising reagents that can chemically react to form products other than the reagents.

[0066] As used herein, "treated polysaccharide" refers to a polysaccharide that has been contacted with at least one reagent capable of reacting with the polysaccharide, such as an enzyme or Fenton's reagent.

[0067] As used herein, "polysaccharide cleavage product" is a product formed by chemical cleavage and / or enzymatic cleavage of a polysaccharide. In some aspects, the polysaccharide cleavage product comprises one or more oligosaccharides. In some aspects, the polysaccharide cleavage product comprises one or more polysaccharides. In some aspects, the polysaccharide cleavage product comprises a mixture of one or more oligosaccharides and one or more polysaccharides.

[0068] As used herein, "oligosaccharide" refers to an oligomer of a carbohydrate, wherein the oligomer has a DP of 2 to 30 monosaccharide units, e.g., 3 to 30, 3 to 20, 3 to 25, 3 to 15, 3 to 10, 3 to 8, 3 to 6, or 5 to 15 monosaccharide units, or any subrange thereof. The oligosaccharide can be linear, branched, primarily linear with side chain sugar monomers, or any combination thereof. "Oligosaccharide" refers to a single oligomer chain.

[0069] As used herein, "oligosaccharide composition" (also referred to herein as "oligosaccharide pool" or "oligosaccharide mixture") refers to a mixture of two or more oligosaccharides, each of which can be the same or different from each other. Although efforts have been made to consistently use the terms "oligosaccharide" and "oligosaccharide composition" according to their aforementioned definitions, when such terms are used herein, the intended meaning will be clear based on the context. In an embodiment, "one or more oligosaccharides" refers to an oligosaccharide mixture when there are more than one oligosaccharide. In an embodiment, "one or more oligosaccharides" refers to an oligosaccharide. In some embodiments, the oligosaccharide composition comprises one or more polysaccharides. According to this aspect, the oligosaccharide composition can comprise up to 60% to 80% by mass of polysaccharides, preferably less than 70% or less than 60% by mass of polysaccharides (e.g., 0.5% to 70% by mass of polysaccharides, 0.5% to 60% by mass of polysaccharides, or 0.5% to 50% by mass of polysaccharides). In preferred embodiments, oligosaccharide compositions comprising up to 60% to 80% by mass polysaccharide have greater solubility, increased bioactivity, or a combination thereof, compared to compositions comprising 80% to 100% by mass polysaccharide.

[0070] As used herein, "subunit" (also sometimes referred to herein as "unit" or "residue") refers to a substance that is covalently bonded to an oligomer (e.g., an oligosaccharide) or a polymer (e.g., a polysaccharide) or covalently bonded to an oligomer (e.g., an oligosaccharide) or a polymer (e.g., a polysaccharide). In some aspects, such a substance may generally include a carbohydrate (e.g., glucose, galactose, mannose, etc.). For example, when an oligosaccharide composition comprises glucose subunits, this means that the composition comprises glucose molecules bound to or bound to an oligomer or polymer; thus, a composition comprising only free monomeric glucose will not comprise glucose subunits. Similarly, when an oligosaccharide composition comprises at least 60% by weight of the total weight of the carbohydrate subunits, this means that the masses of all glucose subunits, galactose subunits, and mannose subunits are added together, and the subunits of all carbohydrates are added together, and then the first sum is divided by the second sum. In addition, when the oligosaccharide composition comprises non-terminal galactose subunits, and it is specified that at least 70% by weight of the non-terminal galactose subunits have at least one 4-bond, the feature is calculated by adding the masses of all non-terminal galactose subunits having at least one 4-bond (and this can include, for example, galactose subunits having 4,6-bonds and galactose subunits having 4,3-bonds) and then dividing by the total mass of the non-terminal galactose subunits regardless of the bond type. The same concept applies to any feature cited herein for "at least one X-bond", where X is an integer (e.g., "the weight ratio of glucose subunits having at least one 4-bond to glucose subunits having at least one 3-bond is 2:1 to 4:1" and other such features). In addition, in this calculation, the actual mass of the subunit is used (i.e., in bound form), rather than the mass of the unit after hydrolysis (which will increase the mass of water). Other features described elsewhere herein can be calculated similarly. These characteristics can be determined using a variety of analytical techniques described herein, such as hydrolysis monosaccharide composition analysis, oligosaccharide analysis, glycosidic bond analysis, NMR HSQC analysis, and the like, as well as other techniques known in the art.

[0071] As used herein, "Fenton's reagent" refers to a reagent comprising a peroxidizing agent and a metal. In certain aspects, the peroxidizing agent is hydrogen peroxide. In certain aspects, the metal is Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II), and Co(II), alkaline earth metal ions Ca(II) and Mg(II), lanthanides Ce(IV), or any combination thereof. In specific embodiments, the metal ion is a copper ion, particularly Cu(II).

[0072] As used herein, the phrase "substantially commensurate with initiation of peroxide quenching" refers to the relationship between the timing of the cleavage reaction and the timing of the peroxide quenching reaction, indicating that initiation of the cleavage reaction and initiation of the peroxide quenching reaction occur within a short period of time of each other (e.g., within about seconds or about minutes, but no more than a day).

[0073] As used herein, "specified reaction time" or "reaction time" refers to the time provided for a reaction to proceed toward a state of equilibrium between the added reagents and the products produced by the reaction of the reagents. In some aspects, the specified reaction time allows sufficient time to reach equilibrium. In certain other aspects, the specified reaction time allows time for the reaction to proceed toward equilibrium but does not provide the time required to reach equilibrium.

[0074] As used herein, the term "synthetic oligosaccharide" refers to the oligosaccharide produced by the depolymerization of one or more polysaccharides. In some aspects, the term synthetic oligosaccharide refers to the composition of the oligosaccharide produced by the method disclosed herein. Alternatively or additionally, enzyme, chemical reaction (such as Fenton chemical reaction), physical process (such as improving time and temperature, etc.) or its any combination can be used to carry out depolymerization to produce synthetic oligosaccharides. In some aspects, the term synthetic oligosaccharide refers to the oligosaccharide prepared by synthesizing oligosaccharides from monosaccharides or lower DP oligosaccharides. The terms "synthetic oligosaccharide" and "oligosaccharide" are used interchangeably in this article, and "composition comprising at least one synthetic oligosaccharide" and "oligosaccharide composition" are used interchangeably in this article. It is intended to be indistinguishable in meaning.

[0075] As used herein, the term "synthetic composition" refers to an artificially prepared composition, and preferably refers to a composition comprising at least one compound produced by an in vitro chemical method and / or biological method (e.g., by chemical reaction, enzymatic reaction, recombinant or any combination thereof). Synthetic compositions typically comprise one or more compounds, including one or more oligosaccharides as described herein. In some aspects, oligosaccharides and oligosaccharide compositions can be formulated into synthetic compositions or administered separately as oligosaccharides. In some embodiments, synthetic compositions can be in the form of a nutritional composition or a pharmaceutical composition.

[0076] As used herein, the term "heteropolymer polysaccharide" refers to a polysaccharide comprising two or more monosaccharide subunits linked together by the same type of glycosidic bonds or different types of glycosidic bonds; heteropolymer polysaccharides also include polysaccharides containing the same type of repeating monosaccharide subunits linked together by different types of glycosidic bonds. The glycosidic bonds in the heteropolymer polysaccharide can be β1-2 bonds, β1-3 bonds, β1-4 bonds, β1-5 bonds, β1-6 bonds, α1-3 bonds, α1-4 bonds, β1-5, α1-6 bonds, or a combination thereof. Examples of heteropolymer polysaccharides include, but are not limited to, xyloglucan, lichenan, β-glucan, glucomannan, galactomannan, arabinan, xylan, and arabinoxylan.

[0077] As used herein, "short-chain fatty acids" include butyrate, propionate, β-hydroxybutyrate, lactate, acetate, or any combination thereof.

[0078] As used herein, the term "hydrolyzed monosaccharide composition analysis" refers to the method described in Amicucci, Galermo et al. 2019 with some modifications, the entire contents of which are incorporated herein by reference for all purposes to the extent not inconsistent with the description herein. In some aspects, the hydrolysis reaction to produce monosaccharides is carried out under optimized conditions at 100 ° C for 2 hours. The sample was run on an Agilent 1290 Infinity II ultra-high performance liquid chromatography (UHPLC) system coupled to an Agilent 6490A triple quadrupole (QqQ) mass spectrometer. Separation was performed on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm × 50 mm, 1.9 μm particle size) plus a guard column (5 mm) using the same solvent system described in Amicucci, Galermo et al. 2019. At a constant flow rate of 1.2 mL / min, an isocratic gradient of 8.5% B was used during the first 4 min elution, followed by elution with 15% B for 0.4 min. During the rinse, 97% B was maintained for 1 min. The column oven was set to 35 ° C. For mass spectrometry parameters, the only change compared to the method described in Amicucci, Galermo et al. 2019 was that the fragmentor voltage was set to 380 V. For data analysis, since the sample contained oligosaccharides rather than polysaccharides, no hydrolysis correction factor was applied and no hydrolysis correction factor was required. In this analytical method, the monosaccharide composition is calculated by quantifying the concentrations of 14 monosaccharides (glucose, galactose, fructose, xylose, arabinose, fucose, rhamnose, glucuronic acid, galacturonic acid, N-acetylglucosamine, N-acetylgalactosamine, mannose, allose, ribose) relative to their respective standard curves. For example, 30% glucose measured by the hydrolysis monosaccharide composition analysis herein means that 30g of glucose is contained in the sum of all 14 monosaccharides mentioned above per 100g.

[0079] As used herein, the term "free monosaccharide composition analysis" refers to the method described in MJ Amicucci et al. 2019 (Amicucci, Galermo et al. 2019) with some modifications. The derivatization reaction to produce monosaccharides was carried out under optimized conditions at 70°C for 30 minutes. The sample was run on an Agilent 1290 Infinity II ultra-high performance liquid chromatography (UHPLC) system coupled to an Agilent 6490A triple quadrupole (QqQ) mass spectrometer. Separation was performed on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm × 50 mm, 1.9 μm particle size) plus a guard column (5 mm) using the same solvent system described in the document. At a constant flow rate of 1.2 mL / min, an isocratic gradient of 8.5% B was used during the first 4 min elution, followed by elution with 15% B for 0.4 min. During the wash, 97% B was maintained for 1 min. The column oven was set to 35°C. For mass spectrometry parameters, compared with the method described in the document, only change is that the fragmentation voltage is set to 380V.For data analysis, because this paper sample comprises oligosaccharides rather than polysaccharides, hydrolysis correction factor is not applied.In this analytical method, by 14 kinds of monosaccharides (glucose, galactose, fructose, xylose, arabinose, fucose, rhamnose, glucuronic acid, galacturonic acid, N-acetylglucosamine, N-acetylgalactosamine, mannose, allose, ribose) are quantitatively calculated inherent free unpolymerized monosaccharide relative to their standard curve separately.For example, 30% free glucose measured by the free monosaccharide composition analysis of this paper refers to that 30g glucose is included in the summation of all 14 kinds of monosaccharides mentioned above of every 100g.

[0080] As used herein, the terms "monosaccharide ratio," "monosaccharide peak area ratio," "monosaccharide ratio," or similar terms may refer to any quantitative comparison based on the relationship observed in the analysis of hydrolyzed monosaccharide composition. The absolute concentration of each monosaccharide is calculated based on the relative percentage relative to the sum of all other monosaccharides observed. The monosaccharide ratio is calculated by dividing the contributing monosaccharide by any other monosaccharide in the composition. The monosaccharide ratio is not intended to limit the composition to the listed monosaccharides. For example, a 1:1 glucose:galactose ratio indicates that approximately equal amounts of glucose subunits and galactose subunits are present in the composition, but the composition may also contain mannose subunits, rhamnose subunits, or any other subunits.

[0081] As used herein, the terms "glycosidic bond composition," "glycosidic bond analysis," "permethylation bond composition analysis," or similar terms refer to the method described in Galermo, Nandita et al. 2018, with some modifications, which is incorporated herein by reference in its entirety for all purposes, to the extent not inconsistent with the description herein. The permethylation reaction time was 30 min. The samples were run on an Agilent 1290 Infinity II UHPLC system coupled to an Agilent 6490A QqQ mass spectrometer. Separation was performed on an Agilent InfinityLab Poroshell HPH-C18 column (2.1 mm × 100 mm, 1.9 μm particle size) plus a guard column (5 mm) using the same solvent system described in Galermo, Nandita et al. 2018. An isocratic gradient of 14% B was used for a 16 min elution period, followed by a 2 min wash period of 99% B at a constant flow rate of 0.8 mL / min. The column oven was set to 35°C. The glycosidic bond composition is calculated by integrating the chromatographic peak areas of all peaks with the following m / z values: 481.2, 495.2, 509.2, 523.3, 525.2, 537.3, 539.3, 553.3, 567.3, 581.3. For example, 20% 4-galactose measured by permethylated bond composition analysis means that the peak area of 4-galactose is 20% of the sum of the peak areas of all peaks with the m / z values listed above.

[0082] As used herein, the term "other minor bonds" refers to the sum of bonds that are not fully annotated or that account for less than 2% of any sample. Therefore, the contribution of these bonds to the glycosidic bond composition of the sample is classified into this "other minor bonds" category.

[0083] As used herein, the term "bond ratio," "bond peak area ratio," "bond ratio," or other similar terms may refer to any quantitative comparison based on the relationship observed in the glycosidic bond composition analysis. For example, an oligosaccharide having a ratio of 1:1 to 1:5 of β-1,3 linked glucose residues: β-1,4 linked glucose residues refers to the bond ratio measured by the glycosidic bond composition analysis. The peak area of each bond is calculated based on the relative percentage of the peak area relative to the sum of the peak areas of all other bonds observed. The peak area ratio is calculated by dividing a contributing bond by any other bond of the same monosaccharide in the composition.

[0084] As used herein, the term "oligosaccharide analysis" or "oligosaccharide composition analysis" (or similar terms) refers to the HPLC-quadrupole time-of-flight (Q-TOF) method described in Amicucci, Nandita et al. 2020 with some modifications, the entire contents of which are incorporated herein by reference for all purposes to the extent not inconsistent with the description herein. For sample preparation, oligosaccharides were reduced by incubating with 2.0M NaBH4 at 65°C for 1 hour. Oligosaccharides were purified using a C-18 column 96-well plate: the plate was washed with 100% ACN, the oligosaccharides were loaded and eluted with water. Oligosaccharides were subsequently purified using a porous graphite carbon (PGC) 96-well plate: the PCG plate was washed with an aqueous solution of 80% acetonitrile and 0.1% (v / v) TFA, the C-18 purified oligosaccharides were loaded and washed with water. The oligosaccharides were eluted with 0.05% (v / v) TFA and 40% acetonitrile. The sample was completely dried by evaporative centrifugation and reconstituted for mass spectrometry analysis. The instrument used was an Agilent 1260 Infinity II HPLC coupled to an Agilent 6530Q-TOF mass spectrometer. Separation was performed using the same stationary phase (plus a 5 mm guard column) and mobile phase as described in the literature, using the following gradient: 2-15% B, 0-20 min; 15-60% B, 20-45 min. The column oven was set to 35°C. The fragmentor voltage was set to 75 V. In this method, when "oligosaccharide weight %" or "oligo wt. %" or similar terms are used in the context of "oligosaccharide analysis," it is calculated by dividing the chromatographic peak area of a specific oligosaccharide by the total peak area of all oligosaccharides determined for that sample during a defined chromatographic period. Generally, when an oligosaccharide composition is described herein as comprising a specified weight percentage, calculated on a dry basis, of oligosaccharides having a specified number of degrees of polymerization (e.g., at least 50% by weight, calculated on a dry basis, of oligosaccharides having a degree of polymerization of 3 to 30 monosaccharide subunits), these values can be calculated using the oligosaccharide analysis described herein; however, other methods may also assist in such determinations, such as size exclusion chromatography using a universal detector, or other methods known in the art.

[0085] As used herein, the term "retention factor" refers to the ratio obtained by dividing the retention time of a given peak observed in an oligosaccharide analysis (eg, HPLC spectrum) by the first oligosaccharide peak (ie, lowest retention time) observed in the oligosaccharide analysis.

[0086] As used herein, the terms "NMR HSQC analysis", "1H-13C HSQC NMR", "HSQC spectrum" or other similar terms correspond to data generated by two-dimensional spectroscopic analysis of a sample by combining heteronuclear single quantum coherence (HSQC) spins of the protons and bonded carbons present in the sample. HSQC experiments rely on dissolution of the sample in a deuterated solvent such as D6-DMSO or D2O. The HSQC spectrum contains a unique peak for each proton associated with the heteronuclear carbon atom under consideration, thereby allowing the identification of the molecular structure of the analyzed sample. Each experiment was performed using a Bruker AVANCE 600MHz NMR using heteronuclear single quantum coherence (HSQC) to illustrate the correlation between the 1H and 13C chemical shifts by 1JCH coupling. The resulting FIDs were processed using Bruker TopSpin 4.1.3 and the experimental chemical shifts were used to determine the anomeric characteristics of the oligosaccharide structure and glycosidic bonds with the aid of the CASPER program. The relative ratios of α and β bonds were calculated by 2D 1H-13C HSQC detection, which measures signal intensities in Hz. These values were then compared to determine the percent abundance of each bond type within the same carbohydrate. The NMR samples were dried by lyophilization and the resulting material was dissolved in 0.75 mL of dimethyl sulfoxide-d6 (DMSO-d6) containing 0.03% (v / v) TMS internal standard at a concentration of 20 mg / mL over a pH range of 4.5-6.

[0087] As used herein, "molecular weight analysis," "molecular weight distribution analysis," or "SEC-RID" or similar terms refer to a method in which samples are prepared by reconstituting a dry powder in nanopure water to make a 10 mg / mL solution. Samples were analyzed on an Agilent Infinity II 1260 RID coupled to an Agilent Infinity II 1260 HPLC. Separation was performed on an Agilent Advance Bio SEC column (7.8 mm x 300 mm, 2.7 μm particle size) with a 10 μL injection volume. The chromatographic solvents were: A: nanopure water, B: 95% acetonitrile in water (v / v), with a gradient of 0.0-13.0 min, 50 min, 0% B; 13.0-14.0 min, 30% B; 14.0-50.0 min, 0% B. The flow rate was set at 1.00 mL / min, and the column temperature was set at 35°C. RID was operated in positive signal polarity mode with a peak width of 2.31 Hz and an attenuation of 500,000 nRIU at 35°C. Samples were integrated using Agilent ChemStation data analysis and molecular weights were determined using an Agilent Cirrus GPC program and a set of Dextran standards with a resolution of 180-150,000 Da. Those skilled in the art will understand that SEC-RID is subject to random experimental error and, therefore, the molecular weights inferred therefrom should be interpreted as including reasonable deviations from the values. Specifically, in some aspects, the molecular weight of the oligosaccharide composition described herein includes a deviation of ±20% of the molecular weight, or in some aspects, a deviation of ±10% of the molecular weight, or in some aspects, a deviation of ±5% of the molecular weight.

[0088] Unless otherwise indicated, the terms "average molecular weight," "average molecular mass," "average molecular weight," "average molecular mass," or similar terms refer to weight average molecular weight. Generally, unless otherwise indicated, when the oligosaccharide or oligosaccharide composition described herein has a specified average molecular weight (e.g., a β-glucan oligosaccharide having a molecular weight distribution such that at least 50% of the mass is less than 5 kDa), these values can be calculated using the molecular weight analysis described herein.

[0089] As used herein, the term "bioactive" or "bioactivity" refers to one or more of the following: reducing glycemic response, reducing Hbalc levels, inhibiting SGLT1 transport, inhibiting α-amylase, inhibiting α-glucosidase, increasing GLP-1 production, enhancing microbial short-chain fatty acid production, enhancing microbial butyrate production, enhancing microbial lactate production, enhancing microbial acetate production, increasing the number of intestinal microbiota members producing short-chain fatty acids, increasing the number of intestinal microbiota members producing butyrate, increasing the number of intestinal microbiota members producing lactate, increasing the number of intestinal microbiota members producing acetate, increasing the abundance of Clostridium butyricum in the intestine, increasing the abundance of Bifidobacterium in the intestine, increasing the abundance of Lactobacillus in the intestine, or increasing the abundance of Lactobacillus rhamnosus in the intestine. In aspects where bioactivity refers to an increase or decrease (e.g., increasing GLP-1 production, reducing Hbalc levels, etc.), as defined herein, an increase or decrease refers to a biologically relevant increase or a biologically relevant decrease.

[0090] As used herein, "increase (enhances or enhancing) microbial production" refers to the biologically relevant increase of the production of a specific metabolite or metabolite group. In some aspects, the biologically relevant increase is a statistically significant change measured by a parametric test or a non-parametric test. In some aspects, the biologically relevant increase can be measured in feces, serum, urine or organ tissue. In some aspects, the biologically relevant increase is measured by host metabolites (choline can be measured by TMA or TMAO). In some aspects, the biologically relevant increase is an increase of 10% or an increase of 100% or an increase of 500% or an increase of 1000% or more. In some aspects, the biologically relevant increase can be the absolute amount of a metabolite or metabolite group. In some aspects, the biologically relevant increase can be the production rate of a metabolite or metabolite group. In some aspects, the biologically relevant increase can be the relative amount of a metabolite or metabolite group.

[0091] As used herein, "reducing microbial production" refers to the biologically relevant reduction of the production of a specific metabolite or metabolite group. In some aspects, the biologically relevant reduction is a statistically significant change measured by a parametric test or a nonparametric test. In some aspects, the biologically relevant reduction can be measured in feces, serum, urine or organ tissue. In some aspects, the biologically relevant reduction is measured by host metabolites (choline can be measured by TMA or TMAO). In some aspects, the biologically relevant reduction is a 10% reduction or a 20% reduction or a 50% reduction or a 75% reduction or a 90% reduction or more. In some aspects, the biologically relevant reduction can be the absolute amount of a metabolite or metabolite group. In some aspects, the biologically relevant reduction can be the production rate of a metabolite or metabolite group. In some aspects, the biologically relevant reduction can be the relative amount of a metabolite or metabolite group.

[0092] As used herein, " reducing microbial utilization " refers to the biologically relevant increase of the amount that causes specific metabolite or metabolite group due to microbial utilization reduction. In some aspects, the biologically relevant increase is the statistically significant change measured by parameter test or non-parametric test. In some aspects, the biologically relevant increase can be measured in feces, serum, urine or organ tissue. In some aspects, the biologically relevant increase is measured by host metabolites (choline can TMA or TMAO). In some aspects, the biologically relevant increase is an increase of 10% or an increase of 100% or an increase of 500% or an increase of 1000% or more. In some aspects, the biologically relevant increase can be the absolute amount of metabolite or metabolite group. In some aspects, the biologically relevant increase can be the production rate of metabolite or metabolite group. In some aspects, the biologically relevant increase can be the relative amount of metabolite or metabolite group.

[0093] As used herein, the term "relative abundance of a bacterium" refers to the abundance of that bacterium relative to other bacteria in the microbiome within or on a particular organ of a complex organism (eg, a human or mammal).

[0094] As used herein, "slowing down microorganism utilization" refers to the biologically relevant increase of the amount or accumulation of a specific metabolite or metabolite group caused by microorganism utilization slowing down. In some aspects, the biologically relevant increase is the statistically significant change measured by parameter test or non-parametric test. In some aspects, the biologically relevant increase can be measured in feces, serum, urine or organ tissue. In some aspects, the biologically relevant increase is measured by host metabolites (choline can be measured by TMA or TMAO). In some aspects, the biologically relevant increase is an increase of 10% or an increase of 100% or an increase of 500% or an increase of 1000% or more. In some aspects, the biologically relevant increase can be the absolute amount of a metabolite or metabolite group. In some aspects, the biologically relevant increase can be the production rate of a metabolite or metabolite group. In some aspects, the biologically relevant increase can be the relative amount of a metabolite or metabolite group.

[0095] As used herein, "increased abundance" refers to a biologically relevant increase in the population of a certain bacterial taxon.

[0096] In some aspects, a "biologically relevant increase" is a statistically significant change measured by a parametric test or a nonparametric test, typically involving the effect of a method comprising administering an oligosaccharide composition or its preparation to a subject, or in vitro, relative to the effect of the identical method not comprising administering an oligosaccharide composition or its preparation. In some aspects, a biologically relevant increase can be measured in feces, jejunum, cecum, ileum, stomach, large intestine, duodenum, oral cavity, respiratory tract, skin, urogenital tract, vagina or other microbial communities. In some aspects, a biologically relevant increase is an increase of 10% or a 5-fold increase or a 10-fold increase or a 50-fold increase or a 100-fold or 1000-fold increase or a 10,000-fold increase or more. In some aspects, a biologically relevant increase can be in terms of the absolute amount of a taxonomic group or taxonomic group, or in terms of the amount of a given substance (e.g., short-chain fatty acids, GLP-1, etc.). In some aspects, a biologically relevant increase can be the rate at which a taxonomic group, taxonomic group or other given substance in a microbial community or subject (or a position therein, e.g., gastrointestinal tract) increases. In some aspects, the increase in biological relevance can be the relative amount of a taxonomic group, a taxonomic group, or other given substance in a microbial community or a subject (or a location therein, such as the gastrointestinal tract). In some aspects, the increase in abundance refers to the presence of a microbial taxonomic group relative to another microbial taxonomic group, or the presence of a given substance relative to another given substance. "Biologically relevant reduction," "biologically relevant change," "biologically relevant amount," and similar such terms can be similarly understood.

[0097] As used herein, "stimulation" with respect to a receptor means that a given substance (eg, butyrate, propionate, etc.) acts as an agonist by binding to the receptor, thereby initiating an immune response.

[0098] As used herein, "increased production" of a given substance (eg, short-chain fatty acids, GLP-1, etc.) means a biologically relevant increase in the production of the given substance.

[0099] As used herein, "inhibiting histone deacetylase" means that the enzymatic activity of the histone deacetylase is reduced by a biologically relevant amount or even eliminated.

[0100] As used herein, "increased expression" of a gene (eg, Muc2, occlding, claudin-4, ZO-1, etc.) refers to a biologically relevant increase in the expression amount of the gene.

[0101] As used herein, "lowering AIc levels" means a biologically relevant decrease in AIc levels.

[0102] As used herein, "reducing a gastrointestinal marker of inflammation" refers to a biologically relevant decrease in the amount of a gastrointestinal marker of inflammation.

[0103] As used herein, "increasing an anti-inflammatory gastrointestinal marker" refers to a biologically relevant increase in the amount of an anti-inflammatory gastrointestinal marker.

[0104] As used herein, "reducing inflammation" in relation to a subject or the gastrointestinal tract of a subject means a reduction in one or more, or any combination of, TNF-α, IL1-β, IL-6, or other known markers of inflammation when compared to the levels of the same markers in a subject that has not undergone the relevant administration or treatment steps using the formulations or oligosaccharide compositions described herein.

[0105] As used herein, "reducing intestinal barrier permeability" in relation to a subject or the gastrointestinal tract of a subject means (1) a decrease in TEER value, and / or (2) an increase in expression of one or more of the Muc2, occluding, claudin-4, ZO-1 genes, or and / or (3) other known intestinal barrier permeability markers, or (4) any combination thereof, when compared to the same markers in a subject that has not undergone the relevant administration or treatment steps using the formulation or oligosaccharide composition described herein.

[0106] "Therapy" means treatment administered or action taken to alleviate or eliminate the symptoms of a disease or pathological condition.

[0107] As used herein, a "therapeutically effective amount" or "effective amount" of a disclosed compound (e.g., an oligosaccharide, oligosaccharide composition, and / or synthetic composition) is a dose of the compound sufficient to achieve a desired therapeutic effect or other result or effect (e.g., an anti-inflammatory effect, stimulation of the growth of a particular microbial population, etc.). For example, a therapeutically effective amount of a compound can be a dose in which a subject receives from about 0.1 μg / kg body weight / day to about 1000 mg / kg body weight / day, for example, from about 1 μg / kg body weight / day to about 1000 μg / kg body weight / day, for example, from about 5 μg / kg body weight / day to about 500 μg / kg body weight / day. One or more compounds herein can be administered single or multiple times, for example, regularly, including once a day, twice a day, every two days, once a week, or once every two weeks, for a specified period of time to achieve and / or maintain a desired therapeutic effect.

[0108] As used herein, "treatment" or "treat" refers to a therapeutic intervention that improves a disease or pathological condition after it has begun to develop, and also includes treating a medical condition or disease for the purpose of improving or stabilizing the results of the subject being treated or solving potential nutritional needs. Therefore, "treatment" or "treat" includes the diet or nutritional management of a medical condition or disease by solving the nutritional needs of the person being treated. "Treating," "treat," and "treatment" have grammatically corresponding meanings. As used herein, the term "improvement" relating to a disease or pathological condition refers to any observable beneficial effect of treatment. Beneficial effect can be demonstrated by, for example, a delayed onset of the clinical symptoms of a disease or condition in susceptible subjects, a reduction in the severity of some or all of the clinical symptoms of a disease or condition, a delay in the progression of a disease or condition, an improvement in the overall health or well-being of the subject, or by other parameters known in the art for a particular disease or condition. The phrases "treating a disease," "treating a condition," and similar terms include inhibiting the full development of a disease or condition, e.g., in a subject at risk for, or in a subject suffering from, a disease or condition, such as inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis, bacterial vaginosis, cardiovascular disease, chronic kidney disease, neurological disorders, allergic reactions, atopic dermatitis, and the like.

[0109] As used herein, "prophylactic treatment" or "prevention" refers to treatment given or action taken to reduce the risk of disease onset or recurrence. Preventing a disease or condition refers to the prophylactic administration of a composition to a subject who does not show signs of the disease or condition or who only shows early signs of the disease or condition, in order to reduce the risk of developing the pathology or condition, or to reduce the severity of the pathology or condition. While in some aspects a disease or condition can be avoided permanently or by retreatment, in other aspects the onset of a disease or condition can be delayed. "Primary prevention" means preventing the first onset of a condition in an individual. "Secondary prevention" means, in a subject who has or has had a condition, (i) preventing the recurrence of the condition, (ii) increasing the duration of remission of the condition, and / or (iii) reducing the severity of the symptoms of the condition. "Preventing," "prevention," "prophylactic treatment," and "prevent" are used interchangeably.

[0110] Emotional disorder refers to a mental disorder that primarily involves emotional imbalance, causing distorted or conflicting emotions. Emotional disorders include excessive anxiety, fear, anger, happiness, etc.

[0111] A mood disorder is a mental disorder that primarily involves dysregulation of emotions, resulting in distorted or conflicted feelings about the environment. Mood disorders include depression, major depressive disorder, dysthymia, and bipolar disorder.

[0112] As used herein, the term "enteral administration" refers to any form of delivery of a composition to a subject that results in deposition of the composition in the gastrointestinal tract (including the stomach). Enteral administration methods include feeding through a nasogastric or jejunal tube, oral feeding, sublingual feeding, and rectal feeding.

[0113] As used herein, the "gastrointestinal tract" or "GI tract" refers to the passage in a subject's digestive system, including all components from the esophagus to the anus, inclusive, and all parts along the passage, including the stomach, intestines, etc. In general, "gastrointestinal tract" is used interchangeably herein with the term "intestine."

[0114] As used herein, the terms "microflota," "microflora," and "microbiome" refer to a community of living microorganisms that typically inhabit an organ or part of the body, such as the gastrointestinal tract or urogenital organs of complex organisms, such as mammals and humans. Specifically, the most important members of the gastrointestinal microbiome include microbial phyla of Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, Synergistetes, Verrucomicrobia, Fusobacteria and Euryarchaeota; microorganisms at the genus level of Bacteroides, Faecalibacterium, Bifidobacterium, Roseburia, Alistipes, Collinsella, Blautia, Coprococcus, Ruminococcus, Eubacterium and Dorea; and microorganisms at the genus level of Bacteroides monomorpha. uniformis, Alistipes putredinis, Parabacteroides merdae, Ruminococcus bromii, Dorea longicatena, Bacteroides caccae, Bacteroides thetaiotaomicron, Eubacterium hallii, Ruminococcus torques, Faecalibacterium prausnitzii, Ruminococcus lactaris, Collinsella aerofaciens, Dorea formicigenerans, Bacteroides vulgatus, and Roseburia intestinalis.The gastrointestinal microbiota includes mucosal-associated microbiota (which resides in or is attached to the mucosal layer covering the gastrointestinal epithelium) and lumen-associated microbiota, which are found within the gastrointestinal lumen. The main members of the urogenital tract microbiota include Lactobacillus crispatus, Lactobacillus jensenii, Lactobacillus gasseri, Lactobacillus iners, and Lactobacillus vaginalis.

[0115] The term "Bifidobacterium" and its synonyms refer to a genus of anaerobic bacteria that are beneficial to humans. Members of the genus Bifidobacterium are some of the major strains that make up the gut microbiome (bacteria that reside in the gastrointestinal tract and have health benefits for their hosts) (Guarner and Malagelada 2003).

[0116] As used herein, the terms "modulate," "modulating," or other similar terms refer to the ability of a disclosed compound (e.g., an oligosaccharide or oligosaccharide composition) to alter the amount, extent, or rate of a biological function (including metabolite production), disease progression, or improvement of a condition. For example, modulation can refer to the ability of a compound to increase or decrease microbial abundance, increase or decrease metabolite production, or cause a decrease in inflammation, pain, morbidity, or severity of symptoms associated with a particular condition or disease (e.g., associated with the gastrointestinal system, cardiovascular system, renal system, nervous system, immune system, and / or genitourinary system).

[0117] As used herein, the term "modulation of the microbiota" refers to exerting a modifying or controlling influence on the microbiota, for example, an influence that results in an increase in the intrinsic intestinal abundance of one or more types of microorganisms (e.g., Bifidobacteria) and / or metabolite-producing bacteria (e.g., butyrate-producing bacteria). In another example, the influence may result in a decrease in the intestinal abundance of one or more types of microorganisms (e.g., Ruminococci and / or Proteobacteria).

[0118] As used herein, the term "oral administration" refers to any form for delivering a composition to a subject through the oral cavity. Thus, oral administration is a form of enteral administration.

[0119] As used herein, the term "EP or ethanol precipitation" refers to a composition artificially prepared by adding a known concentration of ethanol to selectively precipitate and separate the insoluble portion.

[0120] As used herein, the term "ES or ethanol supernatant" refers to a composition artificially prepared by adding ethanol of known concentration to perform selective precipitation and separate the soluble portion.

[0121] Disclosed herein are examples of starting materials (sometimes referred to herein as "material(s)", "materials", "parents", "parent polysaccharides", "precursor polysaccharides" or similar terms) from which certain oligosaccharides and compositions thereof are derived. One skilled in the art will understand that any toxic portions of the starting materials will be minimized or excluded from the methods, compositions, medicaments and formulations herein.

[0122] As used herein, "arabinose" is a polysaccharide whose glycosidic bond composition comprises at least 75% arabinose subunits composed of α-1,3, α-1,5, and α-1,3,5 linkages. In some aspects, arabinose comprises 10-20% galactose subunits, 10-20% xylose subunits, or a combination of 10-20% galactose and xylose subunits. In some aspects, arabinose is "debranched," comprising only α-1,5 arabinose linkages.

[0123] As used herein, "arabinose oligosaccharide" means an oligosaccharide comprising an arabinose residue connected α- and optionally similar to beetroot arabinose and / or legume arabinose (e.g., pea arabinose and / or soybean arabinose). In some aspects, the oligosaccharide comprises an α1-5, α1-3, or α1-2 glycosidic bond. Arabinose oligosaccharides can be linear or branched. In some aspects, the molecular weight distribution of arabinose oligosaccharides is that at least 50% of the mass is less than 50 kDa. As described in WO2021097138A1, WO2018236917A1, WO2020247389A1, and WO2022241163A1, arabinose oligosaccharides can be prepared by Fenton-type depolymerization, and to the extent not contradictory to the description herein, the entire contents of each of these patents are incorporated herein by reference, more specifically citing the synthesis method.

[0124] As used herein, "barley" comprises polysaccharides containing beta-glucans, the glycosidic bond composition of which comprises a glucose backbone containing beta-1,4 and beta-1,3 in a ratio of about 4:1.

[0125] As used herein, "curdlan" is a polysaccharide having a glycosidic bond composition comprising a β-1,3 glucose backbone.

[0126] As used herein, "glucomannan" is a polysaccharide having a glycosidic bond composition with a backbone of about 60% β-1,4 mannose and about 40% β-1,4 glucose.

[0127] As used herein, "xylan" is a polysaccharide having a glycosidic linkage composition comprising a β-1,4 xylose backbone and approximately 13% α-1,2 gluco-4-OMe.

[0128] As used herein, "arabinogalactan" is a polysaccharide having a glycosidic linkage composition comprising a β-1,4 xylose backbone and α-1,3 and α-1,2 arabinose side chains in a ratio of approximately 1:2.

[0129] As used herein, "carob bean gum" is a polysaccharide having a glycosidic bond composition of about 73% β-1,4 mannose backbone, of which about 23% is modified with β-1,4 galactose.

[0130] As used herein, "galactan" is a polysaccharide having a glycosidic bond composition comprising a β-1,4 galactan backbone.

[0131] As used herein, "legumes" refers to any part of a plant of the Fabaceae or Leguminosae family, or the fruit or seeds of such plants. Examples of legumes include peas, beans, soybeans, chickpeas, peanuts, lentils, lupines, mesquite, carob, tamarind, alfalfa, and clover. In some aspects, legumes refer to byproducts of plants during harvest or food processing. Non-limiting examples include powder, pods, flowers, stems, roots, seeds, fiber, or crude protein. Legumes can refer to solid materials that have been extracted by baking, fermentation, hot water, enzymolysis, chemical, alkaline, supercritical fluid, sun-drying, organic solvents, acid, mechanical pressure, or pressure.

[0132] As used herein, "lichenin" is a polysaccharide having a glycosidic bond composition comprising a backbone of β-1,4 glucose alternating with β-1,3 glucose approximately 33% of the time.

[0133] As used herein, "galactomannan" is a polysaccharide having a glycosidic bond composition comprising a β-1,4 mannose backbone and approximately 22% α-1,3 galactose branches.

[0134] As used herein, "β-glucan" (also known as "beta glucan") is a polysaccharide having a glycosidic bond composition comprising a glucose backbone of β-1,4 and β-1,3 in a ratio of about 4:1. In embodiments, the β-glucan has a weight average molecular weight (Mw) of 500 kDa or greater.

[0135] As used herein, "xyloglucan" is a polysaccharide having a glycosidic linkage composition comprising a β-1,4 glucose backbone and α-1,6 xylose side chains.

[0136] As used herein, "arabinoxylan" is a polysaccharide having a glycosidic linkage composition comprising a β-1,4 xylose backbone and α-1,3 and α-1,2 arabinose side chains in a ratio of approximately 1:2.

[0137] As used herein, "olive" refers to any part of the genus Olea. "Olive" can refer to Olea europaea, Olea cuspidate, Olea oleaster, Oleacerasiformis (maderensis), Olea guanchica, Olea laperrinei, Olea maroccana, Olea Canarium or other species. "Olive" can refer to, but is not limited to, green, red, brown or black. "Olive" can refer to the by-products of plants in the harvest or food processing process, non-limiting examples including olive flowers and their related parts (stigma, style, filaments, petals, rachis, articulation and nectar), olive ovules, olive oil, olive oil press cake, olive grinding wastewater, olive pomace, olive compost or olive mud. "Olive" can refer to solid materials extracted through baking, fermentation, hot water, enzymes, chemicals, alkaline processes, supercritical fluids, organic solvents, acid processes, mechanical pressure or pressure. "Olive" may refer to other non-Olea europaea species commonly known as olives.

[0138] As used herein, "Ulvaceae (Ulva intestinalis)" refers to any part of a plant of the genus Ulva, Enteronia, Gemina, Letterstedtia, Lobata, Ochlochaete, Percursaria, Phycoseris, Ruthnielsenia, Solenia, Ulvaria, Umbraulva, or Enteromorpha. "Ulvaceae" may refer to Enteromorpha, Ulva rapa, or other species. "Ulvaceae" may refer to a by-product of a plant during harvest or food processing, non-limiting examples of which include flat or hollow tubular thalluses, Ulvaceae leaves, compound leaves or blades, Ulvaceae strips, or Ulvaceae hold fasts. "Ulva of the Ulva family" may refer to solid materials extracted by roasting, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. "Ulva of the Ulva family" may refer to other non-Ulva species commonly known as sea lettuce, bitter lettuce, or grass kelp.

[0139] As used herein, "macroalgae" refers to any part of the genus Macrocystis. "Macroalgae" may refer to Fucus pyrifer L., Laminaria pyrifera (L.) Lamouroux, Macrocystis humboldtii (Bonpland) C.Ag., Macrocystis planicaulis C.Agardh, Macrocystis pyrifera var. humboldtii, or other species. "Macroalgae" may refer to byproducts of the plant during harvesting or food processing, non-limiting examples of which include flat or hollow tubular thalluses, macroalgae blades, macroalgae air sacs (pneumatocysts), macroalgae strips, macroalgae sporophylls, or macroalgae anchor roots. "Macroalgae" may refer to solid materials that have been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluid methods, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. "Macroalgae" may refer to other non-macroalgae genera commonly known as giant kelp, giant air sac kelp, Pacific kelp, or large brown algae.

[0140] As used herein, "sugar cane" refers to any part of the plant of the genus Saccharum. "Sugar cane" may refer to Saccharum officinarum, Saccharum sinense, Saccharum barberi, Saccharum arundinaceum, Saccharum bengalense, Saccharum edule, Saccharum procerum, Saccharum ravennae, Saccharum robustum, Saccharum spontaneum, hybrids of two, three or more species, or other species. In some aspects, "sugarcane" refers to a byproduct of the plant during harvesting or food processing, non-limiting examples of which include sugarcane leaves (barbojo), sugarcane stalks (stems), raw sugarcane poles or chunks, bagasse, fresh sugarcane juice, sugarcane molasses, raw sugarcane sugar, sugarcane flour, or processed sugarcane. In some aspects, sugarcane refers to a solid material that has been extracted by roasting, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. Sugarcane may also be referred to as "power cane." In some aspects, sugarcane refers to other non-Saccharum species commonly known as sugarcane.

[0141] As used herein, "carrot" refers to any part of the plant of the genus Daucus. "Carrot" may refer to wild carrot (Daucus carota), yellow carrot (Daucus sativus), Carota sativa, or other species. "Carrot" may refer to a by-product of a plant during harvest or food processing, non-limiting examples of which include carrot flowers, carrot stems, carrot seeds, carrot leaves, carrot taproots, or carrot lateral roots. "Carrot" may refer to a solid material that has been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline processes, supercritical fluids, sun drying, organic solvents, acid processes, mechanical pressure, or pressure. "Carrot" may refer to other non-carrot species commonly known as yellow carrot (Daucus carota subsp. Sativus) or wild carrot.

[0142] As used herein, "soy" refers to any part of a plant of the genus Glycine or Soja. "Soy" can refer to Dolichos soja L., Glycine angustifolia Miq., Glycine gracilis Skvortsov, Glycine hispida (Moench) Maxim., Glycine soja, Phaseolus max L., Sojaangustifolia, Soja hispida Moench, Soja japonica Savi, Soja max, Soja H., Sojaviridis, or other species. In some aspects, soy refers to a by-product of a plant during harvest or food processing, non-limiting examples of which include soy roots, soy stems, soy leaves, soy flowers, soy pods, soy seeds, soy protein, soy okra (paste or curd), soy fiber, or soy coat. In some aspects, soybean refers to a solid material that has been subjected to roasting, fermentation, hot water, enzymes, chemicals, alkaline processing, supercritical fluid processing, sun drying, organic solvents, acid processing, mechanical pressure or pressure extraction. "Soybean" can refer to other non-Glycine or non-Soja species commonly known as soy beans, kongbiji or soya.

[0143] As used herein, "Sphingomonas elodea extract" refers to any part of a Sphingomonas bacterium. "Sphingomonas extract" may refer to Pseudomonas elodea or other species. "Sphingomonas" may refer to a by-product of the bacteria during harvesting or food processing, non-limiting examples of which include exopolysaccharides, intracellular polysaccharides, Sphingomonas cell walls, Sphingomonas carbohydrate membranes, or purified Sphingomonas gellan gum polysaccharide. "Sphingomonas extract" may refer to a solid material that has been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure. "Sphingomonas extract" may refer to other non-Sphingomonas bacteria commonly known as gellan gum, bacterial extracts, or gelling agents.

[0144] As used herein, "coffee" refers to any part of the Coffea plant. "Coffee" can refer to Coffea arabica, Coffea robusta, Coffea liberica, or other species. In some aspects, coffee refers to a by-product of a plant during harvest or food processing, non-limiting examples of which include spent coffee grounds, coffee extract, coffee beans, coffee parchment, coffee pulp, coffee berries, coffee fruits, coffee husks, coffee silverskin, coffee gelatin, coffee bean skins, coffee shells, coffee leaves, coffee roots, coffee stems, or coffee leaves. In some aspects, coffee refers to a solid material extracted by roasting, fermentation, hot water, enzymes, chemicals, alkaline processes, supercritical fluids, organic solvents, acid processes, mechanical pressure, or pressure extraction. "Coffee" can refer to other non-Coffee species commonly known as coffee.

[0145] As used herein, "Xanthomonas campestris extract" refers to any part of the Xanthomonas bacteria. The “Xanthomonas campestris extract” may refer to Xanthomonas campestris pv.armoraciae, Xanthomonas campestris pv.begoniae A, Xanthomonas campestris pv.begoniae B, Xanthomonas campestris pv.campestris, Xanthomonas campestris pv.cannabis, Xanthomonas campestris pv.carota, Xanthomonas campestris pv.corylina, Xanthomonas campestris pv.deff dieffenbachiae), Xanthomonas campestris pv. glycines syn, Xanthomonas axonopodis pv. glycines, Xanthomonas campestris pv. graminis, Xanthomonas campestris pv. hederae, Xanthomonas campestris pv. hyacinthi, Xanthomonas campestris pv. juglandis, Xanthomonas campestris pv. malvacearum, or Xanthomonas campestris pv. malvacearum. citrisubsp.malvacearum), Xanthomonas campestrispv.musacearum, Xanthomonas campestrispv.mangiferaeindicae), Xanthomonas campestris pv.mori, Xanthomonas campestris pv.nigromaculans, Xanthomonas campestris pv.pelargonii, Xanthomonas campestris pv.phaseoli, Xanthomonas campestris pv.poinsettiicola, Xanthomonas campestris pv.pruni, Xanthomonas campestris pv.raphani, Xanthomonas campestris pv.sesame In some aspects, the Xanthomonas campestris extract refers to a by-product of the bacteria during harvesting or food processing, non-limiting examples of which include Xanthomonas campestris exopolysaccharides, Xanthomonas campestris intracellular polysaccharides, Xanthomonas campestris cell walls, Xanthomonas campestris carbohydrate membranes, or purified Xanthomonas campestris xanthan polysaccharides. In some aspects, a Xanthomonas campestris extract refers to a solid material extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. "Xanthomonas campestris extract" may also refer to other non-Xanthomonas species commonly known as xanthan gum, bacterial extracts, or gelling agents.

[0146] As used herein, "xanthan gum" refers to a polysaccharide having a β-1,4 glucose backbone alternating with α-1,2 mannose. Xanthan gum can be derived from Xanthomonas campestris. In some aspects, oligosaccharides derived from xanthan gum ("xanthan oligosaccharides") have a variety of structural features similar to the parent polysaccharide. In some aspects, xanthan oligosaccharides include a composition having 63.86% glucose and 30.64% mannose. In some aspects, xanthan oligosaccharides include a glycosidic bond composition of 29.88% 4-glucose, 16.02% 2-mannose, 12.24% 3,6-galactose, 11.54% terminal glucose, and 6.09% 4,6-mannose. In some aspects, xanthan oligosaccharides are produced by enzymatic, chemical, or biosynthesis or by depolymerization of β-glucans by enzymatic, chemical, physical, or biological processes. In some aspects, xanthan oligosaccharides are obtained by Fenton-type depolymerization as described in WO2021097138A1, WO2018236917A1, WO2020247389A1, and WO2022241163A1 (each of which is incorporated herein by reference in its entirety, more particularly with respect to the synthesis methods, to the extent not inconsistent with the disclosure herein). Oligosaccharide CLX123 provides an example of a xanthan oligosaccharide.

[0147] As used herein, "pea" refers to any part of a plant of the genus Pisum, Cajanus, Lathyrus, or Vigina. In some aspects, pea refers to Pisum sativum, Cajanus cajanor, Vigna unguiculata, Lathyrus aphaca, or other species. In some aspects, pea refers to a byproduct of the plant during harvest or food processing, non-limiting examples of which include pea flour, pea pods, pea flowers, pea stems, pea stipules, pea roots, pea seeds, pea fiber, or crude pea protein. In some aspects, pea refers to a solid material that has been extracted by roasting, fermenting, using hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. Peas may refer to other non-Pisum, non-Pigeon, non-Lathyrus, or non-Vigna species commonly known as field peas, snow peas, split peas, sugar peas, garden peas, or sugar peas.

[0148] As used herein, "tomato" refers to any part of the Solanum plant. "Tomato" may refer to Solanum lycopersicum, Lycopersicon lycopersicum, Lycopersicon esculentum, or other species. "Tomato" may refer to a by-product of a plant during harvest or food processing, non-limiting examples of which include tomato skins, tomato berries, tomato stems, tomato flowers, tomato seeds, tomato berry pulp, or tomato roots. "Tomato" may refer to a solid material that has been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. "Tomato" may refer to other non-Lupinus Solanum plants commonly known as tomatoes.

[0149] As used herein, " Saccharomyces cerevisiae " refers to any part of the yeast of Saccharomyces cerevisiae (Saccharomyces) genus. In some aspects, " Saccharomyces cerevisiae " refers to Saccharomyces cerevisiae or other species. Saccharomyces cerevisiae can refer to the by-products of yeast in the harvest or food processing process, and non-limiting examples include yeast cell membranes, yeast growth medium, yeast extracellular polysaccharides, yeast intracellular polysaccharides, yeast cell extracts, yeast fiber, yeast polysaccharides, yeast extracts rich in mannose or yeast spores. In some aspects, Saccharomyces cerevisiae refers to solid materials extracted through baking, fermentation, hot water, enzyme, chemistry, alkali process, supercritical fluid, drying in the sun, organic solvent, acid process, mechanical pressure or pressure-based. In some aspects, Saccharomyces cerevisiae can refer to other non-Saccharomyces cerevisiae that are commonly known as baker's yeast.

[0150] As used herein, "yeast beta glucan" refers to the beta glucan present in the yeast cell wall. In some aspects, yeast beta glucan refers to a polysaccharide comprising beta-linked glucose units, which can be located at the beta-3 position, the beta-4 position, or the beta-6 position. In some aspects, yeast beta glucan exists with other polymers such as mannan. In some aspects, yeast beta glucan refers to a structure in which the backbone is beta-3 linked and the beta-6 bonds are long side chains. In some aspects, yeast beta glucan is derived from Saccharomyces cerevisiae or other yeasts within the genus Saccharomyces or yeasts other than the genus Saccharomyces. In some aspects, yeast beta glucan refers to a solid material that has been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure-based methods.

[0151] As used herein, "β-glucan" (also referred to as "β-glucan") is a polysaccharide comprising β-linked glucose residues. In some embodiments, β-glucan refers to a group of polysaccharides including cereal β-glucans, yeast β-glucans, and fungal β-glucans. In some aspects, the β-glucan polymer comprises β1-3, β1-4, or β1-6 glycosidic linkages. In some aspects, the β-glucan polymer comprises a glycosidic linkage composition comprising a glucose backbone comprising approximately a 4:1 ratio of β-1,4 and β-1,3. In some aspects, the β-glucan is linear or branched. In some aspects, within each class of β-glucans, the distribution of polymers is such that at least 80% by mass is greater than 50 kDa. In some aspects, "β-glucan" refers to a solid material that has been subjected to baking, fermentation, hot water, enzymatic, chemical, alkaline, supercritical fluid, sun drying, organic solvent, acid, mechanical pressure, or pressure extraction. CLX115-PS is an example of a β-glucan.

[0152] As used herein, "β-glucan oligosaccharides" (also referred to as "β-glucan oligosaccharides") refer to a group of oligosaccharides similar to cereal β-glucans, yeast β-glucans, and / or fungal β-glucans. In some aspects, β-glucan oligosaccharides are a class of oligosaccharides comprising only β-linked glucose residues. In some aspects, β-glucan oligosaccharides comprise β1-3, β1-4, or β1-6 glycosidic bonds. In some aspects, β-glucan oligosaccharides are linear or branched. In some aspects, the molecular weight distribution of β-glucan oligosaccharides is such that at least 50% of the mass is less than 5 kDa. In some aspects, "β-glucan oligosaccharides" refer to oligosaccharides produced by enzymatic, chemical, or biological synthesis or by depolymerization of β-glucans by enzymatic, chemical, physical, or biological processes. β-glucan oligosaccharides can be obtained by Fenton-type depolymerization as described in WO2021097138A1, WO2018236917A1, and WO2020247389A1 (each of which is incorporated herein by reference in its entirety, and more particularly with respect to the synthesis methods, to the extent not inconsistent with the present disclosure). Oligosaccharide compositions CLX112, CLX115, and CLX115Cu are all examples of β-glucan oligosaccharides.

[0153] As used herein, "cereal β-glucan" refers to β-glucan present in the cell walls of cereals. "Cereal β-glucan" refers to a polysaccharide comprising β-linked glucose units at the β-3 position and the β-4 position. In some aspects, cereal β-glucan is present with other polymers such as cellulose, starch, and arabinoxylan. In some embodiments, "cereal β-glucan" refers to a structure in which a linear polymer consists of β-4 linked glucose residues interspersed with β-3 linked residues at a ratio of β-4:β-3 linked glucose residues of about 1:1 to 5:1. In some embodiments, cereal β-glucan has a structure in which a linear polymer consists of β-4 linked glucose residues interspersed with β-3 linked residues at a ratio of β-4:β-3 linked glucose residues of about 3:1 to 5:1. Cereal β-glucan can be derived from cereals and grains, such as oats, barley, wheat, rye, and rice. β-glucans can come from other cereals and grains. β-glucans can be extracted from the bran or endosperm of cereals and grains. In some aspects, cereal β-glucans are solid materials extracted by roasting, fermentation, hot water, enzymatic extraction, chemical extraction, alkaline extraction, supercritical fluid extraction, sun drying, extraction with organic solvents, acid extraction, mechanical compression, or pressure extraction. In some embodiments, the distribution of polymers in the cereal β-glucan is such that at least 80% by mass is greater than 50 kDa. CLX115-PS is an example of a cereal β-glucan.

[0154] As used herein, "cereal β-glucan oligosaccharide" refers to an oligosaccharide that is similar to the β-glucan present in the cell walls of cereals and contains β-linked glucose units located at the β-3 position and the β-4 position. In some embodiments, cereal β-glucan oligosaccharides are present in their polysaccharide or oligosaccharide form with polysaccharides or oligosaccharides such as cellulose, starch, and arabinoxylan. In some aspects, cereal β-glucan oligosaccharides refer to a structure in which a linear polymer consists of β-4 linked glucose residues interspersed with β-3 linked residues at a ratio of β-4:β-3 linked glucose residues of about 1:1 to 5:1. In some aspects, cereal β-glucan oligosaccharides have a structure in which a linear polymer consists of β-4 linked glucose residues interspersed with β-3 linked residues at a ratio of β-4:β-3 linked glucose residues of about 3:1 to 5:1. Cereal β-glucan oligosaccharides can be derived from cereal β-glucans from cereals and grains such as oats, barley, wheat, rye, and rice. Cereal β-glucan oligosaccharides can be derived from β-glucans from other cereals and grains. Cereal β-glucan oligosaccharides can be derived from β-glucans extracted from the bran or endosperm of cereals and grains. Cereal β-glucan oligosaccharides can refer to solid materials extracted by roasting, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluid extraction, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. In some aspects, the molecular weight distribution of cereal β-glucan oligosaccharides is such that at least 50% of the mass is less than 5 kDa. In some aspects, cereal β-glucan oligosaccharides refer to oligosaccharides produced by enzymatic, chemical, or biosynthesis, or by depolymerization of β-glucans by enzymatic, chemical, physical, or biological processes. In some aspects, cereal β-glucan oligosaccharides are obtained by Fenton-type depolymerization as described in WO2021097138A1, WO2018236917A1, and WO2020247389A1. Oligosaccharides CLX112, CLX115 and CLX115Cu are all examples of cereal beta-glucan oligosaccharides.

[0155] As used herein, "gum tragacanth" refers to any part of the yeast of the genus Astragalus. "Gum tragacanth" may refer to Astragalus adscendens, Astragalus gummifer, Astragalus brachycalyx, and Astragalus tragacantha, or other species. Gum tragacanth may refer to byproducts of the Astragalus plant during harvesting or food processing, non-limiting examples of which include astragalus juice, astragalus powder, astragalus beans, astragalus leaves, or astragalus bark. "Gum tragacanth" may refer to solid materials that have been extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. Gum tragacanth may refer to other non-Astragalus species commonly known as shiraz gum, shiraz, gumelect, Gond Kateera, or dragon gum.

[0156] As used herein, "orange" refers to any part of the genus Citrus. "Citrus" can refer to grapefruit (Citrus maxima), mandarin orange (Citrus reticulata), sweet orange (Citrus sinensis), sour orange (Citrus aurantium), bergamot (Citrus bergamia Risso), trifoliate orange (Citrus trifoliata) or other different species, variants and hybrids. Citrus can refer to by-products of plants during harvesting or food processing, non-limiting examples of which include citrus peel, citrus pith, citrus pulp, citrus fiber, citrus juice, citrus seeds, citrus leaves, citrus bark or citrus flowers. In some aspects, citrus refers to solid materials extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline processes, supercritical fluids, sun drying, organic solvents, acid processes, mechanical pressure or pressure extraction. "Citrus" can refer to other non-citrus species commonly known as sweet orange, bitter orange, bergamot, trifoliate orange or mandarin orange.

[0157] As used herein, "beet" refers to any part of the plant of the genus Beta. "Beet" can refer to Beta vulgaris or other different species and subspecies adanesisi, maritima, vulgaris, altissima, circla, flavescens, conditiva, and crassa. "Beet" can refer to by-products of the plant during harvest or food processing, non-limiting examples of which include beet taproot, beet stems, beet leaves, beet meal, beet fiber, and beetroot. "Beet" can refer to solid materials that have been roasted, fermented, hot water, enzymatically, chemically, alkaline, supercritical fluid, sun-dried, organic solvent, acid-processed, mechanically pressed, sugared, pickled, or pressure-extracted. "Beet" may refer to other non-beta species commonly known as sugar beet, sea beet, spinach beet, Swiss chard, beetroot, table beet, garden beet, red beet, dinner beet, golden beet, or fodder beet.

[0158] As used herein, "Baobab" refers to any part of a plant of the genus Adansonia. "Baobab" may refer to Adansonia digitata, Adansonia grandidieri, Adansonia gregorii, Adansonia madagascariensis, Adansonia perrieri, Adansonia rubrostipa, Adansonia suarezensis, Adansonia za, or other different species. "Baobab" may refer to a by-product of the plant during harvesting or food processing, non-limiting examples of which include baobab fruit, baobab flour, baobab bark, baobab leaves, baobab fiber, baobab seeds, baobab fruit pith, or baobab flowers. "Baobab" may refer to solid material extracted by roasting, fermentation, hot water, enzymes, chemicals, alkaline methods, supercritical fluids, sun drying, organic solvents, acid methods, mechanical pressure, or pressure extraction. "Baobab" may refer to other non-Adansonia species commonly known as boab, bottle tree, dead rat tree, monkey-bread tree, or montane.

[0159] As used herein, "Karayu gum" refers to any part of a plant of the genus Sterculia. "Karayu gum" may refer to Sterculia urens, Cavallium urens, Clompanus urens, Kavalama urens, or other different species. "Karayu gum" may refer to a by-product of the plant during harvesting or food processing, non-limiting examples of which include karaya gum liquid, karaya gum powder, karaya gum leaves, or karaya bark. "Karayu gum" may refer to a solid material extracted by baking, fermentation, hot water, enzymes, chemicals, alkaline processes, supercritical fluids, sun drying, organic solvents, acid processes, mechanical pressure, or pressure extraction. "Karayu gum" may refer to other non-Karayu gum species commonly known as gum tragacanth, katira, kulu, or gum sterculia.

[0160] As used herein, "lupin galactan" refers to any part of the Lupinus plant. "Lupin" can refer to yellow lupin (Lupinus arboreus), blue lupin (Lupinus hirsutus), white lupin (Lupinus chamissonis), Campanulaceae lupin (Lupinus albifrons), Lupinus excubitus, white lupin (Lupinous albus), Lupinous mutabilis, long-leaf lupin (Lupinus longifolius), narrow-leaf lupin (Lupinous angustifolius), or other different species. "Lupin galactan" can refer to a by-product of the plant during harvesting or food processing, non-limiting examples of which include lupin beans, lupin flour, lupin seeds, lupin flowers, lupin stems, "protein-extracted" lupin, lupin fiber, and defatted lupin flour. "Lupin galactan" may refer to a solid material extracted by roasting, fermentation, hot water, enzymatic, chemical, alkaline, supercritical fluid, sun drying, organic solvent, acid, mechanical pressure or pressure extraction. "Lupin" may refer to other non-Lupinus species commonly known as lupin beans, white lupin, tarwi, chocho, kirku, turmus or blue lupin.

[0161] As used herein, when a table, spectrum or other data is referred to as representing a characteristic or property possessed by a particular composition, oligosaccharide or other compound or mixture, unless otherwise indicated, the same analytical methods and procedures used to obtain the table, spectrum or other data will be used to determine the property of the particular composition, oligosaccharide or other compound or mixture.

[0162] Various compositions disclosed herein are identified by the designation "CLX". CLX generally refers to an oligosaccharide composition. In embodiments where CLX is followed by "PS", CLX#-PS refers to a polysaccharide composition. Such CLX compositions can be prepared in any suitable manner and by any suitable method, including de novo synthesis methods (e.g., oligomerizing monomers or shorter chain oligosaccharides into the oligosaccharides shown), or by depolymerization methods (e.g., by depolymerizing polysaccharides or longer chain oligosaccharides into shorter chain oligosaccharides). For example, in some aspects, the CLX compositions disclosed herein can be prepared by the depolymerization methods disclosed in WO 2018 / 236917 (Amicucci et al., "Production of bioactive oligosaccharides") or WO 2021 / 097138 (Amicucci et al., "High-yield peroxide quench-controlled polysaccharide depolymerization and compositions thereof"), the entire contents of which are incorporated herein by reference for all purposes, more particularly with respect to methods of preparation, to the extent not inconsistent with the description herein. For example, the CLX compositions disclosed herein can be prepared by a method comprising dissolving a polysaccharide of a specified origin (e.g., microbial curdlan, lichenin, xylan, etc.) in 20 mL of HPLC-grade water in a covered reaction vessel and placing in a shaking incubator at 55° C. and 85 RPM for 20 minutes. The pH of the solution is adjusted to 5.2 or about 5.2. Hydrogen peroxide (5 mL) and iron (II) sulfate or copper (II) sulfate (2.75 mg in 50 μL of water, in either case) are added to the reaction mixture and mixed thoroughly. The reaction in the covered reaction vessel is carried out in a shaking incubator at 55° C. and 65 RPM for two hours. The sealed reaction vessel is cooled to 12° C. in a −20° C. refrigerator. The pH is adjusted using ammonium hydroxide (1 mL of 28% v / v, to a pH of 8 to 12, e.g., 10.2), and the sample is reacted in a shaking incubator at 45° C. and 20 RPM for 1 hour, with the lid loosened to allow the release of oxygen, ammonia, and carbon dioxide gases. Alternatively, sodium hydroxide (65 μ L 10.45M NaOH to pH 8 to 12, for example 10) can be used instead of ammonium hydroxide. The sample is then frozen and lyophilized, then stored at -80 ℃. The freeze-dried oligosaccharide mixture is rehydrated with the minimum amount of water required for allowing the solution to flow freely. The solution is then loaded onto a post that every gram (dry weight) of raw material comprises 15 mL of mixed bed ion exchange resin, and the effluent is collected in a plastic freezing bag. Once the material is loaded onto the post, the post is rinsed with 3 times of bed volumes of water. Finally, the effluent is sealed and frozen in a bag, then carefully broken and lyophilized.In embodiments, the depolymerization process is performed using established production methods, such as batch or continuous processes.

[0163] For NMR analysis of the CLX compositions described herein, oligosaccharides were dissolved in D2O or D6-DMSO at a concentration of 50 mg / mL and their HSQC spectra were analyzed on a 600 MHz Bruker NMR spectrometer.

[0164] The oligosaccharide compositions disclosed herein (including CLX compositions) are characterized, in part, by the relative amounts of monosaccharide subunits present in each composition. The amount of each subunit is expressed as a percentage as defined herein under "Hydrolysis Monosaccharide Composition Analysis" and / or as a "Monosaccharide Ratio" as defined herein. Those skilled in the art will understand that hydrolysis monosaccharide composition analysis is subject to random experimental error, and therefore percentages and ratios should be interpreted as including reasonable deviations from the stated values. Specifically, in some aspects, percentages and ratios relating to the monosaccharide subunits of an oligosaccharide composition include a deviation of ±20% of the stated percentage or ratio. According to this aspect, for a CLX composition described as comprising 50% of its mass containing glucose, the skilled artisan will understand that the CLX composition may comprise glucose at any percentage between 40% and 60% of its mass. As a further example in this respect, the glucose of 2: 1: galactose ratio includes the glucose of 1.6: 1 to 2.4: 1 (for example, 1.6: 1, 1.8: 1, 2: 1, 2.2: 1, 2.4: 1) and 2: 0.8 to 2: 1.2 (for example, 2: 0.8, 2: 0.9, 2: 1, 2: 1.1, 2: 1.2): galactose ratio deviation. In some aspects, the percentage and ratio relevant to the monosaccharide subunits of oligosaccharide composition include the deviation of ± 10% of the percentage or ratio. In some aspects, the percentage and ratio relevant to the monosaccharide subunits of oligosaccharide composition include the deviation of ± 5% of the percentage or ratio. In some aspects, the percentage and ratio relevant to the monosaccharide subunits of oligosaccharide composition include the deviation of ± 1% of the percentage or ratio.

[0165] Relatedly, oligosaccharide compositions (including CLX compositions) are also characterized by the relative amount of glycosidic bonds present in each composition. The amount of each bond is expressed as a percentage as defined herein under "glycosidic bond composition" and / or as a "bond ratio" as defined herein. Those skilled in the art will understand that glycosidic bond composition analysis is subject to random experimental error, and therefore, percentages and ratios should be interpreted as including reasonable variations in the values. Specifically, in some aspects, the percentages and ratios associated with the glycosidic bonds of the oligosaccharide composition include a deviation of ±20% of the percentage or ratio. In some aspects, the percentages and ratios associated with the glycosidic bonds of the oligosaccharide composition include a deviation of ±10% of the percentage or ratio. In some aspects, the percentages and ratios associated with the glycosidic bonds of the oligosaccharide composition include a deviation of ±5% of the percentage or ratio. In some aspects, the percentages and ratios associated with the glycosidic bonds of the oligosaccharide composition include a deviation of ±1% of the percentage or ratio.

[0166] "Diabetes" is a disease characterized by hyperglycemia and a relative lack of insulin. Diabetes is diagnosed when fasting blood sugar is ≥7.0mmol / L (126mg / dL), or when blood sugar ≥11.1mmol / L (200mg / dL) 2 hours after administration in a glucose tolerance test. There are three main types, namely type 1 diabetes, type 2 diabetes and gestational diabetes. Type 1 diabetes is an autoimmune disease that results from the destruction of insulin-producing beta cells by the immune system. Type 2 diabetes is caused by a combination of lifestyle and genetic factors, leading to hyperglycemia, insulin resistance, and ultimately beta cell damage. Gestational diabetes refers to high blood sugar levels that occur during pregnancy in women who do not have diabetes. In this specification, unless a specific form is specified, the term diabetes includes all three forms of the disease.

[0167] "Obese" means a body mass index (BMI) exceeding 30 kg / m 2 For a human individual, BMI is a measurement obtained by dividing an individual's weight by the square of their height.

[0168] "Overweight" means a body mass index (BMI) range of 25-30 kg / m 2 For a human individual, BMI is a measurement obtained by dividing an individual's weight by the square of their height.

[0169] As used herein, the terms "patient" or "subject" generally refer to a living organism suffering from or susceptible to a disease or condition that can be treated by administering a compound or pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammals. In some embodiments, the patient is a human. In aspects where the patient is a human, the patient can be a child or adult patient. In some embodiments, the patient is a mammal. In some embodiments, the patient is a mouse. In some embodiments, the patient is a laboratory animal. In some embodiments, the patient is a rat. In some embodiments, the patient is a test animal.

[0170] "Prediabetes" means that an individual has at least one of the following characteristics: a glycated hemoglobin (A1C) level of 5.7% to 6.4%, a fasting blood glucose level of 100 mg / dL to 125 mg / dL (5.6 mmol / L to 7.0 mmol / L), or a blood glucose level of 140 mg / dL to 199 mg / dL (7.8 mmol / L to 11.0 mmol / L).

[0171] "At risk for diabetes" means having risk factors that indicate a higher risk of developing diabetes. Non-limiting examples of risk factors include: a waist circumference greater than 100 cm for men or greater than 88 cm for women, blood pressure of 130 / 85 mmHg or higher, a blood triglyceride level greater than 150 mg / dl, a fasting blood glucose level greater than 100 mg / dl, a high-density lipoprotein level less than 40 mg / dl for men, or a high-density lipoprotein level less than 50 mg / dl for women. An individual "at risk for diabetes" may have one or more of these factors.

[0172] A "synthetic composition" refers to an artificially prepared composition and optionally refers to a composition comprising at least one compound produced by in vitro chemical and / or biological methods (e.g., by chemical reaction, enzymatic reaction, or recombination). Synthetic compositions typically comprise β-glucan oligosaccharides. In addition, in some embodiments, the synthetic composition may include one or more nutritionally active ingredients or pharmaceutically active ingredients that do not adversely affect the effects of the β-glucan oligosaccharides. Some non-limiting embodiments of synthetic compositions are described below. DETAILED DESCRIPTION

[0173] In the following description, many specific details of the oligosaccharides, oligosaccharide compositions and methods of the present invention are set forth to provide a comprehensive explanation of the precise nature of the present invention. However, it will be apparent to those skilled in the art that the present invention can be practiced without these specific details. Although the following description is divided into multiple sections, it is contemplated that each section comprises multiple aspects of the present invention, and disclosures within each section and across two or more sections may be combined to form any aspect of the present invention.

[0174] In this specification, it will be shown that by depolymerizing the polysaccharide β-glucan fiber, the oligosaccharide units may have an enhanced inhibitory effect on amylase, α-glucosidase, and SGLT1. Without wishing to be bound by any particular theory, it is believed that this increased effect may be due to reduced substrate steric hindrance, more frequent interactions between proteins and oligosaccharides, and reduced solution viscosity, thereby increasing the availability of oligosaccharides to the enzyme / transporter active sites. In addition, the oligosaccharide units are more easily digested by the intestinal microbiota, resulting in an increase in the concentration of short-chain fatty acids in the colon. β-glucan oligosaccharides are also easier to formulate than the original polysaccharide and have better organoleptic properties, improving subject compliance.

[0175] In some aspects, a method for treating a glucose-related metabolic disorder in a subject is provided. In embodiments, glucose-related metabolic disorders include diabetes (type 1 diabetes, type 2 diabetes, or gestational diabetes) or metabolic syndrome. In embodiments, a method for reducing a subject's postprandial glucose response is provided. In some embodiments, the subject is at risk of developing diabetes, is overweight and / or obese, is pregnant, and / or has diabetes. In some embodiments, a method for reducing the risk of prediabetes subjects and / or obese subjects progressing to type 2 diabetes is provided. In some embodiments, a method for reducing a subject's HbA1c level is provided. In the above embodiments, the method includes administering an effective amount of β-glucan oligosaccharides to the subject enterally before and / or during the subject's intake of a glucose source. In the method for reducing a subject's HbA1c level, the subject's postprandial glucose response is weakened over a period of at least 2 months by enteral administration. In embodiments of the above methods, β-glucan oligosaccharides are administered up to 2 hours before the subject ingests the glucose source. In embodiments of the above methods, β-glucan oligosaccharides are administered up to 1 hour before the subject ingests the glucose source. In embodiments of the above methods, the beta-glucan oligosaccharide is administered up to 30 minutes before the subject ingests the glucose source. In embodiments of the above methods, the beta-glucan oligosaccharide is administered up to 15 minutes before the subject ingests the glucose source.

[0176] In embodiments, an effective amount of β-glucan oligosaccharide for treatment ranges from about 0.5 g to about 20 g. In embodiments, an effective amount of β-glucan oligosaccharide ranges from about 0.75 g to about 15 g, such as about 1 g to about 7.5 g.

[0177] The present disclosure provides several β-glucan oligosaccharides and methods for their use. In embodiments, the β-glucan oligosaccharide inhibits salivary amylase or pancreatic amylase, and / or the β-glucan oligosaccharide inhibits SGLT1 glucose transporter, and / or the β-glucan oligosaccharide inhibits α-glucosidase.

[0178] In an embodiment, the β-glucan oligosaccharide comprises β-1,3 and β-1,4 linked glucose residues. In an embodiment, the β-glucan oligosaccharide comprises both β-1,3 and β-1,4 linked glucose residues. In an embodiment, the β-glucan oligosaccharide comprises a ratio of 1:1 to 1:5 of β-1,3 linked glucose residues: β-1,4 linked glucose residues, for example 1:1, 1:2, 1:3, 1:4 or 1:5. In an embodiment, the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 10,000 Da, optionally greater than 500 Da or 1,000 Da. In an embodiment, the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 8,000 Da, optionally greater than 500 Da or 1,000 Da. In an embodiment, the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 7,500 Da, optionally greater than 500 Da or 1,000 Da. In an embodiment, the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 5,000Da, optionally greater than 500Da or 1,000Da. In an embodiment, the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 2,500Da, optionally greater than 500Da or 1,000Da. In an embodiment, the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue. In an embodiment, the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue. In an embodiment, the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein each subunit is a β-1,3 glucose residue or a β-1,4 glucose residue, and the β-glucan oligosaccharide comprises both β-1,3 glucose residue and β-1,4 glucose residue. In embodiments, the β-glucan oligosaccharide comprises 3 to 30 subunits, or 3 to 25 subunits, or 5 to 30 subunits or 5 to 25 subunits, or 10 to 30 subunits or 10 to 25 subunits, or any subrange thereof.

[0179] In an embodiment, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C. In an embodiment, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C. In an embodiment, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C. In an embodiment, the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C. In an embodiment, the β-glucan oligosaccharide has a dynamic viscosity of about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C.

[0180] In embodiments, at least 70% by mass of the β-glucan oligosaccharides have a molecular mass of less than 100 kDa, optionally greater than 0.1 kDa, 0.5 kDa, or 1 kDa. In embodiments, at least 60% by mass of the β-glucan oligosaccharides have a molecular mass of less than 50 kDa, optionally greater than 0.1 kDa, 0.5 kDa, or 1 kDa. In embodiments, at least 50% by mass of the β-glucan oligosaccharides have a molecular mass of less than 15 kDa, optionally greater than 0.1 kDa, 0.5 kDa, or 1 kDa. In embodiments, at least 50% by mass of the β-glucan oligosaccharides have a molecular mass of less than 5 kDa, optionally greater than 0.1 kDa, 0.5 kDa, or 1 kDa. In embodiments, at least 25% by mass of the β-glucan oligosaccharides have a molecular mass of less than 1 kDa, optionally greater than 0.1 kDa.

[0181] In an embodiment, the solubility of the β-glucan oligosaccharide is 50 mg / mL to 1000 mg / mL, e.g., 50 mg / mL to 1000 mg / mL, 100 mg / mL to 1000 mg / mL, 150 mg / mL to 1000 mg / mL, 200 mg / mL to 1000 mg / mL, 150 mg / mL to 500 mg / mL, or 200 mg / mL to 1000 mg / mL. In a preferred embodiment, the solubility of the β-glucan oligosaccharide is at least 200 mg / mL, optionally less than 1000 mg / mL. In a further preferred embodiment, the solubility of the β-glucan oligosaccharide is at least 200 mg / mL, optionally less than 1000 mg / mL, and the turbidity value is less than 20 NTU, optionally greater than 0.5 NTU.

[0182] In various aspects, the compositions disclosed herein (including CLX compositions) have a turbidity value of 0.5 NTU to 50 NTU, e.g., 0.5 NTU to 50 NTU, 0.5 NTU to 40 NTU, 0.5 NTU to 30 NTU, 0.5 NTU to 25 NTU, or 0.5 NTU to 20 NTU. In preferred embodiments, the compositions disclosed herein (including CLX compositions) have a turbidity value of 20 NTU, optionally greater than 0.5 NTU, optionally greater than 1 NTU.

[0183] In an embodiment, the present disclosure provides a method for producing an oligosaccharide composition, in particular a method for producing the β-glucan oligosaccharide of the present disclosure, and for the uses described herein. In an embodiment, the β-glucan oligosaccharide is produced by the following steps: reacting a polysaccharide in a reaction mixture with a Fenton reagent having a peroxidase agent and a metal ion to provide a treated polysaccharide; and lysing the treated polysaccharide with an alkali to produce a polysaccharide lysate and / or a mixture of oligosaccharides having polysaccharide characteristics, which mixture is a β-glucan oligosaccharide. In an embodiment, the Fenton reagent comprises hydrogen peroxide and one or more metal ions selected from the following: transition metals Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II) and Co(II), alkaline earth metals Ca(II) and Mg(II), and lanthanide element Ce(IV). In embodiments, the base is one or more bases selected from the group consisting of ammonium hydroxide, ammonium bicarbonate, ammonia, urea, sodium amide, dimethylamine, trimethylamine, pyridine and N,N-diisopropylethylamine, sodium hydroxide, calcium hydroxide, potassium hydroxide, barium hydroxide, lithium hydroxide. In embodiments, the base is one or more bases selected from the group consisting of ammonium hydroxide and sodium hydroxide. In embodiments, the base is a nitrogen-based cleavage reagent. In embodiments, the nitrogen-based cleavage reagent is also a peroxide quenching reagent, and the initiation of polysaccharide cleavage is synchronized or substantially synchronized with the initiation of peroxide quenching. In embodiments, the nitrogen-based cleavage reagent is not a peroxide quencher, and the method further comprises initiating peroxide quenching with an additional peroxide quencher. In embodiments, wherein the metal ion is a copper ion. In embodiments, the metal ion is Cu(II). In embodiments, the concentration of Cu(II) used in the reaction mixture is about 0.25 mM to about 1.00 mM. In embodiments, the concentration of Cu(II) used in the reaction mixture is about 0.7 mM to about 0.8 mM. In embodiments, the concentration of Cu(II) used in the reaction mixture is about 0.75 mM. In embodiments, the source of Cu(II) is copper sulfate. In embodiments, the Fenton reagent comprises copper sulfate. In embodiments, the hydrogen peroxide concentration is about 1% to about 7%. In embodiments, the hydrogen peroxide concentration is about 3.5% (v / v) to 4.5% (v / v). In embodiments, the hydrogen peroxide concentration is about 4.0 (v / v). In embodiments, the alkali concentration is about 0.2M to about 1M. In embodiments, the alkali concentration is about 0.3M to about 0.5M. In embodiments, the alkali concentration is about 0.4M. In embodiments, the alkali is added to a final pH of about 8 to 12. In embodiments, the alkali is added to a final pH of about 8.5 to 11. In embodiments, the alkali is added to a final pH of about 9.5 to 10.5. In embodiments, the alkali is added to a final pH of about 10. In embodiments, the polysaccharide loading in the reaction mixture is about 2% (w / v) to about 20% (w / v).In embodiments, the polysaccharide loading in the reaction mixture is about 8% (w / v) to about 12% (w / v).In embodiments, the polysaccharide loading in the reaction mixture is about 10% (w / v).

[0184] In an embodiment, the polysaccharide used to generate the beta-glucan oligosaccharide is derived from cereal grains. In an embodiment, the polysaccharide used to generate the beta-glucan oligosaccharide is derived from oats or barley. In an embodiment, the polysaccharide used to generate the beta-glucan oligosaccharide is a beta-glucan, particularly a beta-glucan having a weight average molecular weight of 500 kDa or greater, optionally less than 10,000 kDa. In some embodiments, the polysaccharide used to generate the beta-glucan oligosaccharide is derived from a by-product or waste stream of oats or barley, such as milk solids of oats or barley, brewers spent grain, or pearled barley dust.

[0185] In an embodiment, β-glucan is purified from the starting material by dry grinding and sorting. In an embodiment, β-glucan is purified by wet grinding and sorting with centrifugation, a decanter, a tri-canter centrifuge, and a hydrocyclone. In an embodiment, β-glucan is purified by removing starch by amylase digestion (with or without jet cooking or other steam cooking techniques). In an embodiment, β-glucan is purified by removing protein by protease digestion. Suitable proteases include flavor enzymes, alkaline proteases, papain, bromelain, protease A, protease B, neutral proteases, trypsin, chymotrypsin, pepsin, and subtilisin. Enzyme hydrolysates (e.g., glucose, maltose, maltooligosaccharides, peptides, and amino acids) can be removed from β-glucan by ethanol precipitation or common solid-liquid separation techniques (decanter, tri-canter centrifuge, centrifuge, nutsche dryer), or by size exclusion methods such as membrane filtration, terminal filtration, or chromatography techniques.

[0186] On the other hand, the present disclosure provides the use of the beta-glucan oligosaccharides described herein for treating glucose-related metabolic disorders, or the use of the beta-glucan oligosaccharides described herein for preparing a medicament for treating glucose-related metabolic disorders. In an embodiment, the use is for treating diabetes or metabolic syndrome. In an embodiment, the use is for treating type 1 or type 2 diabetes or gestational diabetes. In an embodiment, the use is for reducing the postprandial glucose response of a subject. In an embodiment, the subject is at risk of developing diabetes, is overweight and / or obese, is pregnant and / or has diabetes. In an embodiment, the use is for reducing the risk of prediabetic subjects and / or obese subjects progressing to type 2 diabetes. In an embodiment, the use is for reducing the HbA1c level of a subject.

[0187] In another aspect, the present disclosure provides a pharmaceutical composition comprising the β-glucan oligosaccharide described herein and a pharmaceutically acceptable carrier.

[0188] In yet another aspect, the present disclosure provides a dietary supplement comprising the β-glucan oligosaccharide described herein and a food-grade carrier.

[0189] In some aspects, a method for controlling glucose levels by applying an oligosaccharide composition generated from cereal grain beta-glucans is provided. In some aspects, the oligosaccharide composition is generated from oats, barley, rye, or wheat. In some aspects, the oligosaccharide composition is generated from beta-glucans derived from cereals. In some aspects, the oligosaccharide composition can disrupt the breakdown of starch. In some aspects, the oligosaccharide composition can disrupt the breakdown of maltooligosaccharides and maltose into glucose. In some aspects, the oligosaccharide composition can disrupt the absorption of glucose from the intestine. In some aspects, the oligosaccharide composition can inhibit salivary amylase and pancreatic amylase. In some aspects, the oligosaccharide composition can inhibit alpha-glucosidase. In some aspects, the oligosaccharide composition can inhibit the SGLT1 transporter.

[0190] In some aspects, the oligosaccharide composition can be used to treat prediabetes. In some aspects, the oligosaccharide composition can be used to treat type 1 diabetes. In some aspects, the oligosaccharide composition can be used to treat type 2 diabetes. In some aspects, the oligosaccharide composition can be used to treat gestational diabetes. In some aspects, the oligosaccharide composition can be used to reduce postprandial blood sugar spikes. In some aspects, the oligosaccharide composition can be used to lower HbA1c levels.

[0191] In some aspects, the oligosaccharide composition can be produced by depolymerizing β-glucans using a Fenton system. In some aspects, the Fenton system uses iron and / or copper. In some aspects, the composition can be produced by enzymatic depolymerization. In some aspects, the enzymatic depolymerization uses lichenase, β-glucanase and / or cellulase. In some aspects, the oligosaccharide composition is produced by algae, yeast and / or bacterial fermentation. In some aspects, the oligosaccharide composition is produced by chemical synthesis or chemoenzymatic synthesis. In some aspects, the oligosaccharide composition is produced by autolysis. In some aspects, the oligosaccharides are produced by genetic engineering. In some aspects, the oligosaccharides are extracted from food.

[0192] In some aspects, the oligosaccharide composition is fermented by intestinal microflora. In some aspects, the oligosaccharide composition stimulates the production of short-chain fatty acids. In some aspects, the short-chain fatty acids are butyrate or propionate or acetate or beta-hydroxybutyrate or lactate. In some aspects, the interaction between the oligosaccharide composition and the intestinal microflora can help control blood sugar levels. In some aspects, short-chain fatty acids can stimulate the release of GLP-1. In some aspects, GLP-1 stimulates insulin secretion. In some aspects, the oligosaccharide composition works synergistically by first inhibiting amylase, α-glucosidase and SGLT1 in the small intestine and then fermenting by the microflora in the small intestine and colon.

[0193] In some aspects, oligosaccharide comprises β1-3 and β1-4 glucose glycosidic bond. In some aspects, oligosaccharide library or at least one oligosaccharide in library comprises the β1-3 of about 0.37:1 ratio: β1-4 glucose glycosidic bond. In some aspects, oligosaccharide library or at least one oligosaccharide in library comprises the β1-3 of about 0.20:1 to about 0.37:1 ratio: β1-4 glucose glycosidic bond. In some aspects, when measured by SEC-RID, at least 70% of the mass is less than 100kDa, optionally greater than 0.1kDa or greater than 0.5kDa. In some aspects, when measured by SEC-RID, at least 60% of the mass is less than 50kDa, optionally greater than 0.1kDa or greater than 0.5kDa. In some aspects, when measured by SEC-RID, at least 50% of the mass is less than 15kDa, optionally greater than 0.1kDa or greater than 0.5kDa. In some aspects, when measured by SEC-RID, at least 50% of the mass is less than 5kDa, optionally greater than 0.1kDa or greater than 0.5kDa. In some aspects, when measured by SEC-RID, at least 25% of the mass is less than 1kDa, optionally greater than 0.1kDa or greater than 0.5kDa. In some aspects, the oligosaccharide library comprises oligosaccharides having a DP scope of 3 to 30.

[0194] In this specification, unless otherwise specified, the following terms have the following meanings:

[0195] As used herein, the term "CLX101" refers to an oligosaccharide composition of which approximately 99% by mass comprises glucose (as measured by hydrolysis monosaccharide composition analysis). For composition CLX101, the glycosidic bond composition comprises approximately the amounts of CLX101 listed in Table B (75% 3-linked glucose, 9% terminal glucose, and 15% other minor bonds). The CLX101 composition comprises approximately the 1H-13C HSQC NMR correlations for CLX101 listed in Table A. The CLX101 composition comprises approximately the values measured by oligosaccharide analysis listed in Table C. CLX101 has a dynamic viscosity of 1.306 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX101 is derived from a microbial curdlan. In some aspects, CLX101 is generally derived from microbial gellan, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX101. CLX101 is produced by depolymerization as described in Example 7.

[0196] As used herein, the term "CLX102" refers to an oligosaccharide composition wherein 37% by mass comprises glucose and 60% by mass comprises mannose (as measured by hydrolysis monosaccharide composition analysis). For composition CLX102, the glycosidic bond composition comprises approximately the amounts listed in Table B for CLX102 (32% 4-linked glucose, 8% terminal glucose, 48% 4-linked mannose, and 13% terminal mannose). The CLX102 composition comprises approximately the 1H-13C HSQC NMR correlations for CLX102 listed in Table A. The CLX102 composition comprises approximately the values listed in Table D for oligosaccharide analysis. CLX102 has a dynamic viscosity of 1.392 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX102 is typically derived from konjac glucomannan, but can also be derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX102. CLX102 is produced by depolymerization as described in Example 7.

[0197] As used herein, the term "CLX112" refers to an oligosaccharide composition wherein 97% by mass comprises glucose (as measured by hydrolyzed monosaccharide composition analysis). For composition CLX112, the glycosidic bond composition comprises approximately the amounts listed in Table B for CLX112 (17% 3-linked glucose, 49% 4-linked glucose, and 31% terminal glucose). The CLX112 composition comprises approximately the 1H-13C HSQC NMR correlations for CLX112 listed in Table A. The CLX112 composition comprises approximately the values listed in Table E as measured by oligosaccharide analysis. The molecular weight distribution of the CLX112 composition comprises approximately the values listed in Table K (as measured by refractive index detection (RID)) (see also Figure 7D ). CLX112 has a dynamic viscosity of 1.248 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX112 is typically derived from barley beta glucan, but can also be derived from other materials / sources (e.g., polysaccharide depolymerization or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX112. In one aspect, CLX112 is produced according to the depolymerization described in Example 7.

[0198] As used herein, the term "CLX113" refers to an oligosaccharide composition comprising 49% by mass of glucose, 36% by mass of xylose, and 14% by mass of galactose (as measured by hydrolysis monosaccharide composition analysis). For composition CLX113, the glycosidic bond composition comprises approximately the amounts of CLX113 listed in Table B (28% 4-linked glucose, 6% 6-linked glucose, 20% 4,6-linked glucose, 4% terminal glucose, 21% terminal galactose, 6% 2-linked xylose, and 11% terminal xylose). The CLX113 composition comprises approximately the 1H-13 CHS QC NMR correlations for CLX113 listed in Table A. The CLX113 composition comprises approximately the values measured by oligosaccharide analysis listed in Table F. CLX113 has a dynamic viscosity of 1.209 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX113 is generally derived from tamarind seed xyloglucan. In some aspects, CLX113 is derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX112. In one aspect, CLX113 is produced by depolymerization as described in Example 7.

[0199] As used herein, the term "CLX115" refers to an oligosaccharide composition wherein 95% by mass comprises glucose and 2% by mass comprises arabinose (as measured by hydrolysis monosaccharide composition analysis). For composition CLX115, the glycosidic bond composition comprises approximately the amounts of CLX115 listed in Table B (64% 4-linked glucose, 23% 3-linked glucose, and 13% terminal glucose). The CLX115 composition comprises approximately the 1H-13C HSQC NMR correlations for CLX115 listed in Table A. The CLX115 composition comprises approximately the values listed in Table G as measured by oligosaccharide analysis. The molecular weight distribution of the CLX115 composition comprises approximately the values listed in Table K (as measured by refractive index detection (RID)) (see also Figure 7C). CLX115 has a dynamic viscosity of 1.382 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX115 is generally derived from oat beta glucan. In some aspects, CLX115 is derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX115. In one aspect, CLX115 is produced according to the depolymerization described in Example 7.

[0200] As used herein, the term "CLX123" refers to an oligosaccharide composition wherein 64% by mass comprises glucose, 31% by mass comprises mannose, and 3% by mass comprises glucuronic acid (as measured by hydrolysis monosaccharide composition analysis). For composition CLX123, the glycosidic bond composition comprises approximately the amounts listed in Table B for CLX123 (30% 4-linked glucose, 3% 3-linked glucose, 12% terminal glucose, 12% 3,6-linked galactose, 16% 2-linked mannose, 6% 4,6-linked mannose, 6% terminal mannose, and 4% 4-linked glucuronic acid). The CLX123 composition comprises approximately the 1H-13C HSQC NMR correlations for CLX123 listed in Table A. The CLX123 oligosaccharide composition comprises approximately the values listed in Table H as measured by oligosaccharide analysis. CLX123 has a dynamic viscosity of 1.658 mPa*s at 100 mg / mL at 25°C. In some aspects, CLX123 is generally derived from an extract of Xanthomonas campestris. In some aspects, CLX123 is derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and H-C HSQC NMR analysis as CLX123. In one aspect, CLX123 is produced according to the depolymerization described in Example 7.

[0201] As used herein, the term "CLX125" refers to an oligosaccharide composition comprising 44% by mass of glucose, 43% by mass of rhamnose, and 8% by mass of glucuronic acid (as measured by hydrolysis monosaccharide composition analysis). For composition CLX125, the glycosidic bond composition comprises approximately the amounts of CLX125 listed in Table B (24% 4-linked glucose, 21% 3-linked glucose, 10% terminal glucose, 22% 4-linked rhamnose, 7% terminal rhamnose, 5% 4-linked glucuronic acid, and 3% terminal glucuronic acid). The CLX125 composition comprises approximately the 1H-13CHS QC NMR correlations for CLX125 listed in Table A. The CLX125 oligosaccharide composition comprises approximately the values measured by oligosaccharide analysis listed in Table I. CLX125 has a dynamic viscosity of 1.525 mPa*s at 25°C at 100 mg / mL. In some aspects, CLX125 is generally derived from a Sphingomonas extract. In some aspects, CLX125 is derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX125. In one aspect, CLX125 is produced by depolymerization as described in Example 7.

[0202] As used herein, the term "CLX115-PS" (OatBG poly) refers to a polysaccharide composition wherein 99% by mass comprises glucose (as measured by hydrolysis monosaccharide composition analysis). For the composition CLX115-PS, the glycosidic bond composition comprises approximately the amounts listed in Table B for CLX115-PS. The molecular weight distribution of CLX115-PS comprises approximately the values listed in Table K (as measured by refractive index detection (RID)) (see also Figure 7A In one aspect, the oat β-glucan polysaccharide of CLX115-PS is sourced from PurestarChem and is 90% pure. Oat β-glucan is reported to have a molecular weight ranging from 65,000 Da to 3,100,000 Da (doi:10.3390 / ijms20164032).

[0203] As used herein, the term "CLX112-PS" (BarleyBG poly) refers to a polysaccharide composition wherein 97% by mass comprises glucose (as measured by hydrolysis monosaccharide composition analysis). For the composition CLX112-PS, the glycosidic bond composition comprises approximately the amounts listed in Table B for CLX112-PS. In one aspect, the barley β-glucan polysaccharide of CLX112-PS is sourced from Megazyme Inc. (Product No. P-BGBH). Barley β-glucan is reported to have a molecular weight ranging from 31,000 Da to 2,700,000 Da (doi:10.3390 / ijms20164032).

[0204] As used herein, the term "CLX115Cu" refers to an oligosaccharide composition wherein 87.5% by mass comprises glucose and 4.5% by mass comprises arabinose (as measured by hydrolysis monosaccharide composition analysis). For the composition CLX115Cu, the glycosidic bond composition comprises approximately the amounts of CLX115Cu listed in Table B. The CLX115Cu oligosaccharide composition comprises approximately the values listed in Table J as measured by oligosaccharide analysis. The molecular weight distribution of the CLX115Cu composition comprises approximately the values listed in Table K (as measured by refractive index detection (RID)) (see also Figure 7B In some aspects, CLX115Cu is generally derived from oat β-glucan. In some aspects, CLX115Cu is derived from other materials / sources (e.g., depolymerization of polysaccharides or oligomerization of lower DP monosaccharides and / or oligosaccharides) that provide oligosaccharides having the same (or substantially the same, e.g., within 10%, or within 15%, or within 20%, or within 25%, or within 30%) dynamic viscosity, hydrolyzed monosaccharide composition, glycosidic bond composition, oligosaccharide analysis, and 1H-13C HSQC NMR analysis as CLX115Cu. In one aspect, CLX115Cu is produced according to the depolymerization described in Example 6.

[0205] Table A: 1H-13C HSQC NMR correlations for oligosaccharide compositions used herein. The peak pairs listed correspond to those major peaks in the anomeric region.

[0206]

[0207] Table A (continued)

[0208]

[0209] Table B. Glycosidic bond analysis of CLX compositions. Data are presented as peak area %. "Other" refers to bonds that contribute less than 2%. The symbol "--" indicates a bond present in an amount less than 2% (and may be 0%) of the total oligosaccharide weight. If the bond is not fully described, the bond is represented by the monosaccharide (when known) or monosaccharide type (pentose or hexose), followed by multiple or single "x"s to indicate the number of branch points.

[0210] Blank cell = not measured / no signal

[0211]

[0212]

[0213] Table C: CLX101 contains the oligosaccharides shown in this table. Hex refers to hexose, Pent refers to pentose, HexA refers to hexadronate, and Deoxyhex refers to deoxyhexose.

[0214]

[0215] Table D: CLX102 contains the oligosaccharides shown in this table. Hex refers to hexose, Pent refers to pentose, HexA refers to hexauronic acid sugar, and Deoxyhex refers to deoxyhexose.

[0216]

[0217]

[0218]

[0219] Table E: CLX112 contains the oligosaccharides shown in this table. Hex refers to hexose, Pent refers to pentose, HexA refers to hexauronic acid, and Deoxyhex refers to deoxyhexose.

[0220]

[0221]

[0222] Table F: CLX113 contains the oligosaccharides shown in this table. Hex refers to hexose, Pent refers to pentose, HexA refers to hexauronic acid sugar, and Deoxyhex refers to deoxyhexose.

[0223]

[0224]

[0225] Table G: CLX115 contains the oligosaccharides shown in this table. Hex refers to hexose, Pent refers to pentose, HexA refers to hexauronic acid sugar, and Deoxyhex refers to deoxyhexose.

[0226]

[0227]

[0228]

[0229] Table H: CLX123 contains the oligosaccharides indicated in this table (Pent = pentose, typically arabinose, Hex = hexose, typically glucose or mannose).

[0230]

[0231]

[0232]

[0233] Table I. CLX125 contains the oligosaccharides shown in this table.

[0234]

[0235]

[0236] Table J. CLX115Cu contains the oligosaccharides shown in this table.

[0237]

[0238]

[0239]

[0240] Table K. Refractive Index Detection (RID) Analysis of CLX Compositions. Data are presented in units of peak area %.

[0241]

[0242] In one embodiment, the present invention provides a method for reducing a subject's postprandial glucose response. The method comprises administering an effective amount of β-glucan oligosaccharide to a subject before and / or during the subject's intake of a glucose source. The β-glucan oligosaccharide can be provided in the form of a synthetic composition, such as a formulation, supplement, nutritional composition, or medicament of substantially pure β-glucan oligosaccharide.

[0243] In one embodiment, the synthetic composition is a dietary supplement in unit dosage form comprising a unit dose of beta-glucan oligosaccharides. The dietary supplement may comprise an acceptable food grade carrier, such as a phosphate buffered saline solution, a mixture of ethanol and water, water and an emulsion (e.g., oil / water or water / oil emulsion) and a variety of wetting agents or excipients. The dietary supplement may also comprise other materials that do not produce adverse reactions, allergic reactions or other unwanted reactions when administered to a human. The carrier and other materials may include solvents, dispersants, coatings, absorption enhancers, controlled release agents and one or more inert excipients, such as starch, granulating agents, microcrystalline cellulose, diluents, lubricants, binders and disintegrants.

[0244] The dietary supplement can be administered orally, for example, as a tablet, capsule or granule containing a predetermined amount of beta-glucan oligosaccharide, or as a powder or granule containing a predetermined amount of beta-glucan oligosaccharide, or as a gel, paste, solution, suspension, emulsion, syrup, pill, electuary or slurry containing a predetermined amount of beta-glucan oligosaccharide in an aqueous or non-aqueous liquid. Orally administered compositions can include one or more binders, lubricants, inert diluents, flavorings and wetting agents. Orally administered compositions such as tablets can optionally be coated and can be formulated to provide sustained, delayed or controlled release of the beta-glucan oligosaccharide.

[0245] Dietary supplements can also include activating agents, such as vitamins, mineral substances, prebiotics, probiotics and anti-inflammatory agents. The example of suitable vitamins includes vitamin A, B complex vitamins (such as B1, B2, B6 and B12), vitamin C, vitamin D, vitamin E and vitamin K, niacin and acidic vitamins such as pantothenic acid, folic acid and biotin. The example of suitable mineral substances includes calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium and boron. The example of suitable prebiotics includes human milk oligosaccharides, galacto-oligosaccharides, fructo-oligosaccharides, inulin etc. Suitable probiotics include B. animalis subsp. lactis BB-12, B. lactis HN019, B. lactis Bi07, B. infantis ATCC 15697, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus HN001, Lactobacillus acidophilus LA-5, Lactobacillus acidophilus NCFM, Lactobacillus fermentum CECT5716, B. longum BB536, B. longum AH 1205, B. longum ...NCFM, Lactobacillus fermentum CECT5716, B. longum BB536, B. longum NCFM, Lactobacillus fermentum CECT5716, B. longum NCFM, Lactobacillus fermentum CECT5716, B. longum BB536, B. longum NCFM, Lactobacillus fermentum CECT5716, B. longum NCFM, Lactobacillus fermentum CECT5716, B. longum BB536, B. longum NCFM, Lactobacillus fermentum CECT5 1205), Bifidobacterium longum AH1206 (B. longum AH1206), Bifidobacterium breve M-16V (B. breve M-16V), Lactobacillus reuteri ATCC 55730 (L reuteri ATCC 55730), Lactobacillus reuteri ATCC PTA-6485 (L reuteri ATCC PTA-6485) and Lactobacillus reuteri DSM 17938 (L reuteri DSM 17938). Suitable anti-inflammatory agents include natural antioxidants and phenolics such as carotenoids, for example, lutein, lycopene, zeaxanthin and beta-carotene. In addition, fats with anti-inflammatory properties such as omega-3 polyunsaturated fatty acids can be included.

[0246] The dietary supplement may also include one or more other agents suitable for use in foods and dietary supplements that improve glucose management. Suitable agents are known in the art. Non-limiting examples include alpha-lipoic acid, chromium, magnesium, vanadium, cinnamon bark extract, berberine extract, fenugreek seed extract, gymnema extract, mulberry leaf extract, aloe extract, turmeric / curcumin extract, raspberry extract, bitter melon extract, ginseng extract, or any combination thereof.

[0247] The resulting composition may also be formulated in unit dosage form for administration via a nasogastric tube or direct infusion into the gastrointestinal tract or stomach.

[0248] In a further embodiment, the synthetic composition can be formulated as a pharmaceutical composition. The pharmaceutical composition can include a pharmaceutically acceptable carrier, such as a phosphate buffered saline solution, a mixture of ethanol and water, water and an emulsion (such as oil / water or water / oil emulsion) and a variety of wetting agents or excipients. The pharmaceutical composition can also be included in other materials that do not produce adverse reactions, allergic reactions or other unnecessary reactions when applied to a person. The carrier and other materials can include solvents, dispersants, coatings, absorption enhancers, controlled release agents and one or more inert excipients, such as starch, granulating agents, microcrystalline cellulose, diluents, lubricants, binders and disintegrants.

[0249] The pharmaceutical composition can also include one or more other agents suitable for improving the medicine of glucose management. Suitable agents are known in the art. Non-limiting examples include metformin, sulfonylureas (Glyburide, Glipizide, Glimepiride), glinides (Repaglinide, Nateglinide), thiazolidinediones (Pioglitazone, Rosiglitazone), DPP-4 inhibitors (Sitagliptin, Saxagliptin), and saxagliptin.

[00145] The present invention relates to a combination of angiotensin-converting enzyme inhibitor (IgA1c), angiotensin-converting enzyme inhibitor (IgE1c), angiotensin-1 (IgE1c), angiotensin-2 (IgE2), angiotensin-3 (IgE1), angiotensin-4 (IgE2), angiotensin-5 (IgE1), angiotensin-6 (IgE2), angiotensin-7 (IgE1), angiotensin-8 (IgE2), angiotensin-12 (IgE2), angiotensin-13 (IgE1), angiotensin-14 (IgE2), angiotensin-13 ...

[0250] The pharmaceutical composition can be orally administered, for example, as a tablet, capsule or granule comprising a predetermined amount, or as a powder or granule comprising a predetermined amount, or as a gel, paste, solution, suspension, emulsion, syrup, pill, lozenge or slurry comprising a predetermined amount, in an aqueous or non-aqueous liquid. Orally administered compositions can include binding agents, lubricants, inert diluents, flavorings and wetting agents. Orally administered compositions such as tablets can optionally be coated and can be formulated to provide a continuous, delayed or controlled release of a mixture thereof.

[0251] The pharmaceutical composition may also be administered via a nasogastric tube or infused directly into the gastrointestinal tract or stomach.

[0252] Pharmaceutical compositions may also contain therapeutic agents such as antibiotics, probiotics, analgesics, and anti-inflammatory agents.

[0253] The synthetic composition can be in the form of a nutritional composition. For example, the nutritional composition can be a food composition, a rehydration solution, a medical food or a food for special medical purposes, a nutritional supplement, etc. The nutritional composition can contain a protein, a lipid and / or a digestible carbohydrate source and can be in powder or liquid form. The composition can be designed as a sole source of nutrition or as a nutritional supplement.

[0254] Suitable protein sources include milk protein, soy protein, rice protein, pea protein and oat protein, or mixtures thereof. Milk protein can be in the form of milk protein concentrate, milk protein isolate, whey protein or casein, or a mixture of the two. Protein can be whole protein or hydrolyzed protein (partially hydrolyzed or extensively hydrolyzed). Hydrolyzed protein provides the advantage of being easier to digest, which is very important for people with inflamed or damaged gastrointestinal tract. Protein can also be provided in the form of free amino acids. Protein can comprise about 5% to about 30%, typically about 10% to 20%, of the energy of the nutritional composition. The protein source can be glutamine, threonine, cysteine, serine, proline or a combination of these amino acids.

[0255] Suitable digestible carbohydrates include maltodextrin, starch or corn starch, glucose polymers, corn syrup, corn syrup solids, high fructose corn syrup, rice-derived carbohydrates, pea-derived carbohydrates, potato-derived carbohydrates, tapioca, sucrose, glucose, fructose, sucrose, lactose, honey, sugar alcohols (such as maltitol, erythritol, sorbitol) or their mixtures. Preferably, in the composition, glucose is reduced or not added or the simple digestible carbohydrates comprising glucose are included. Conventionally digestible carbohydrates provide approximately 15% to approximately 55% of the nutritional composition energy, for example, approximately 35% to approximately 55%. Particularly suitable digestible carbohydrates are low glucose equivalent (DE) maltodextrins.

[0256] Suitable lipids include medium chain triglycerides (MCT) and long chain triglycerides (LCT). Typically, lipids provide about 15% to about 50% of the nutritional composition's energy, such as about 30% to about 50%. Lipids can include essential fatty acids (ω-3 fatty acids and ω-6 fatty acids). Preferably, these polyunsaturated fatty acids provide less than about 30% of the lipid source's total energy.

[0257] Suitable sources of long-chain triglycerides are rapeseed oil, sunflower oil, palm oil, soybean oil, butterfat, corn oil, high oleic oil and soy lecithin. Fractionated coconut oil is a suitable source of medium-chain triglycerides. The lipid profile of the nutritional composition is preferably designed to have a ratio of polyunsaturated fatty acids ω-6 (n-6) to ω-3 (n-3) of about 4:1 to about 10:1. For example, the ratio of n-6 fatty acids to n-3 fatty acids can be about 6:1 to about 9:1 (by weight).

[0258] The nutritional composition can also include vitamins and minerals. If the nutritional composition is used as the sole nutritional source, it preferably includes a complete vitamin and mineral spectrum. Examples of vitamins include vitamin A, B complex vitamins (such as B1, B2, B6, and B12), vitamin C, vitamin D, vitamin E, and vitamin K, niacin, and acidic vitamins such as pantothenic acid, folic acid, and biotin. Examples of minerals include calcium, iron, zinc, magnesium, iodine, copper, phosphorus, manganese, potassium, chromium, molybdenum, selenium, nickel, tin, silicon, vanadium, and boron.

[0259] The nutritional composition can also include carotenoids, such as lutein, lycopene, zeaxanthin, and beta-carotene. The total amount of carotenoids included can range from about 0.001 pg / mL to about 10 pg / mL. The amount of lutein included can be from about 0.001 pg / mL to about 10 pg / mL, preferably from about 0.044 pg / mL to about 5 pg / mL of lutein. The amount of lycopene included can be from about 0.001 pg / mL to about 10 pg / mL, preferably from about 0.0185 pg / mL to about 5 pg / mL of lycopene. Beta-carotene can include from about 0.001 pg / mL to about 10 mg / mL, such as from about 0.034 pg / mL to about 5 pg / mL of beta-carotene.

[0260] The nutritional composition preferably also contains a reduced concentration of sodium; for example, about 300 mg / L to about 400 mg / L. The remaining electrolytes may be present at concentrations sufficient to meet needs without unduly burdening renal solutes on renal function. For example, potassium is preferably present in a range of about 1180 mg / L to about 1300 mg / L; chloride is preferably present in a range of about 680 mg / L to about 800 mg / L.

[0261] The nutritional compositions may also contain a variety of other conventional ingredients, such as preservatives, emulsifiers, thickeners, buffering agents, fiber and prebiotics, probiotics, antioxidant / anti-inflammatory compounds including tocopherols, carotenoids, ascorbate / vitamin C, ascorbyl palmitate, polyphenols, glutathione and superoxide dismutase (melon), other bioactive factors (e.g., growth hormones, cytokines, TFG-b), colorants, flavorings and stabilizers, lubricants, etc.

[0262] The nutritional composition may also include one or more other agents suitable for use in foods and dietary supplements that improve glucose management. Suitable agents are known in the art. Non-limiting examples include alpha-lipoic acid, chromium, magnesium, vanadium, cinnamon bark extract, berberine extract, fenugreek seed extract, gymnema extract, mulberry leaf extract, aloe vera extract, turmeric / curcumin extract, raspberry extract, bitter melon extract, ginseng extract, or any combination thereof.

[0263] The nutritional composition can be formulated as a soluble powder, a liquid concentrate, or a ready-to-use formulation. The composition can be administered to a person in need thereof via a nasogastric tube or orally. Various flavorings, fibers, and other additives may also be present.

[0264] Nutritional composition can be prepared by any common manufacturing technology for preparing the nutritional composition of solid or liquid form. For example, composition can be prepared by combining multiple feed solutions (feed solution). Feed solutions with protein in fat can be prepared by heating and mixing lipid sources, then adding emulsifiers (such as lecithin), fat-soluble vitamins and at least a portion of protein sources when heating and stirring. Then, carbohydrate feed solutions are prepared by adding minerals, trace and ultra-trace minerals, thickeners or suspending agents in water when heating and stirring. Before adding carbohydrates (such as HMO and digestible carbohydrate sources), the solution obtained is continued to be heated and stirred for 10 minutes. Then, the feed solutions obtained are blended when heating and stirring, and pH is adjusted to 6.6-7.0, then the composition is subjected to high temperature short-time processing, during which time, the composition is subjected to heat treatment, emulsification and homogenization, and then cooled. Add water-soluble vitamins and ascorbic acid, if necessary, pH is adjusted to the desired range, add flavorings, and add water to reach the desired total solids level.

[0265] For liquid products, the resulting solution can then be aseptically packaged to form an aseptically packaged nutritional composition. In this form, the nutritional composition can be in a ready-to-eat or concentrated liquid form. Alternatively, the composition can be spray-dried and processed and packaged as a resolvable powder.

[0266] In order to weaken the subject's postprandial glucose response, the amount of β-glucan oligosaccharide administered to the subject will vary according to factors such as risk factors, the severity of the subject's condition, the subject's age, the form of the composition, the glucose load and type of the food / beverage consumed, and other drugs being administered to the subject. However, the amount can be easily set by a physician. In several embodiments, the dosage range for weakening the postprandial glucose response can generally be about 0.5g to about 20g, for example, about 0.75g to about 15g, more specifically, about 1g to about 7.5g. β-glucan oligosaccharide can be administered before or when the patient consumes a glucose source. When administered before consuming a glucose source, β-glucan oligosaccharide can be administered up to about 30 minutes before, for example, up to about 15 minutes before. When administered when consuming a glucose source, β-glucan oligosaccharide can be administered separately from the glucose source or as a meal replacement. The daily dose of β-glucan oligosaccharide administered to the subject is generally about 0.5g / day to about 20g / day. For example, the daily dose can be about 0.75g / day to about 15g / day, more specifically, about 1g / day to about 10g / day. Suitable dosage can be determined according to several factors, including, for example, body weight and / or disease, severity of disease, other ailments and / or diseases, incidence and / or severity of side effects, mode of administration and / or glucose load of the diet / beverage taken in. Suitable dosage ranges can be determined by methods known to those skilled in the art.

[0267] In order to reduce the risk of prediabetic subjects and / or obese subjects progressing to type 2 diabetes, the amount of β-glucan oligosaccharide administered to the subject will vary according to factors such as risk factors, the severity of the subject's condition, the subject's age, the form of the composition, the glucose load and type of the food / beverage taken, and other drugs being administered to the subject. However, the amount can be easily set by a physician. The dosage and daily dose suggested above can be used. β-glucan oligosaccharide can be administered daily over a period of time (e.g., at least one month, at least two months, etc.).

[0268] The amount of β-glucan oligosaccharide administered to a subject to lower their HbA1c level will vary depending on factors such as risk factors, the severity of the subject's condition, the subject's age, the form of the composition, the glucose load and type of food / beverage consumed, and other medications being administered to the subject. However, the amount can be readily determined by a physician. The dosages and daily doses suggested above can be used.

[0269] The resulting composition can be co-administered to patients who are also receiving standard of care medications for glucose control.

[0270] Example

[0271] Example 1: Evaluation of the inhibitory effect of β-glucan oligosaccharides on SGLT1 receptor

[0272] It has been described that some polysaccharides not only reduce SGLT-1 expression in the intestinal mucosa of diabetic mice, but also reduce SGLT-1 expression in Caco-2 cells. In addition, some long carbohydrate structures significantly promote blood glucose regulation in normal mice and reduce glucose transport through Caco-2 monolayers (Cao et al. 2016). One method for measuring SGLT1-mediated transport is the cellular uptake assay. This interaction is detected by adjusting the initial rate of transport of 14C-AMG by human SGLT1 to HEK-293-FRT cells expressing the SGLT1 transporter. Inhibition experiments were performed using phlorizin (100 μM) as a reference inhibitor to ensure maximum inhibition of SGLT1-mediated AMG transport.

[0273] β-glucan oligosaccharides were produced using isolated oat β-glucan as described in Example 8. Two different concentrations of β-glucan oligosaccharides (300 μM and 3000 μM) were tested in the assay. When β-glucan oligosaccharides were used, a dose-dependent SGLT1 inhibition was observed, with a relative transporter inhibition of 14% at 3000 μM ( Figure 1 These results support the use of β-glucan oligosaccharides as an agent for reducing glucose uptake and thereby minimizing postprandial glucose levels.

[0274] Example 2: Evaluation of α-glucosidase inhibition. α-glucosidase is located in the brush border of the small intestine and is responsible for the enzymatic degradation of starch, producing glucose as one of the main products. α-glucosidase is a target for regulating postprandial hyperglycemia, helping to lower postprandial blood glucose levels by preventing glucose absorption in the intestine. It has been reported that carbohydrate digestibility is associated with elevated postprandial blood glucose. One strategy to reduce postprandial hyperglycemia is to limit the activity of carbohydrate-digesting enzymes in the intestine. α-Amylase is an enzyme that degrades polymeric substrates into shorter oligomers by catalyzing the hydrolysis of α-1,4-glucan bonds in starch, maltodextrin, and other related carbohydrates (Truscheit et al., 2010).

[0275] The inhibitory effect of CLX115 on α-glucosidase was measured using BioVision's α-glucosidase inhibitor screening kit. This kit exploits the ability of active α-glucosidase to cleave synthetic substrates, thereby releasing a chromophore. In the presence of a specific α-glucosidase inhibitor, reduced enzyme activity is detected by a decrease in absorbance readings. Using acarbose as a positive inhibition control, multiple concentrations of nine carbohydrate sources were tested (Table L). In separate experiments testing only CLX115, reactions included 100 mg / mL CLX115 or 200 μM acarbose, 20 μM maltose, and 2 U / mL of α-glucosidase. Multiple test tubes were prepared for each treatment and incubated at 37°C. The progress of the reaction was monitored by placing the test tubes in a 96°C water bath for 10 minutes to inactivate the enzyme and stopping the reaction at 0, 10, 30, and 60 minutes. Glucose and maltose were measured by the method of Xu et al. (10.1039 / C7AN01530E). In brief, glucose and maltose were derivatized with PMP (3-methyl-1-phenyl-2-pyrazolidine-5-one) and analyzed on an Agilent UHPLC / QqQ mass spectrometer. Quantification was performed using a standard curve and peak area was normalized using an internal standard.

[0276] The results showed that CLX115, CLX112, and CLX115Cu inhibited α-glucosidase at the tested concentrations, whereas various other carbohydrate sources (including CLX115-PS) showed no inhibitory effect (Table L, Figure 2 and Figure 3 For a separate experiment testing only CLX115, when α-glucosidase was incubated with maltose alone, the results showed that the maltose level decreased as the reaction proceeded ( Figure 8A ), free glucose accumulation ( Figure 8B CLX115 and acarbose (positive control) both inhibited the release of glucose ( Figure 8B ) and inhibit the decomposition of maltose ( Figure 8A ). In fact, in the presence of 100 mg / mL CLX115, maltose and glucose levels remained almost unchanged until the end of the reaction. This indicates that CLX115 effectively inhibits α-glucosidase activity. This inhibitory effect suggests that CLX115 can be used as an agent to reduce the postprandial blood glucose increase. The inhibitory effect of CLX 115-PS and its oligosaccharide counterparts on α-glucosidase was evaluated. However, turbidity and viscosity of the polysaccharide solution are important factors that may serve as confounding variables. This inhibitory effect suggests that depolymerized cereal β-glucans from various sources can be used as agents to reduce glucose uptake. Table L. α-glucosidase inhibition results. NA = Not measured, X = Not inhibited

[0277]

[0278] Example 3: In vitro evaluation of short-chain fatty acid production by CLX115.

[0279] Short-chain fatty acids (especially acetate, propionate, and butyrate) are primarily produced by anaerobic fermentation of intestinal microorganisms. SCFAs have been shown to have beneficial physiological effects, such as stimulating G protein-coupled receptors. Studies have shown that SCFAs activate GPR41, which can stimulate the secretion of the intestinal hormone glucagon-like peptide-1 (GLP-1). GLP-1 indirectly regulates blood glucose levels by increasing insulin secretion and reducing glucagon secretion.

[0280] An existing intestinal model, the Human Intestinal Microbial Ecosystem Simulator (SHIME), was used to evaluate the effect of CLX115 on SCFA production. This system allows for the simulation of the physiology and microbiology of the gastrointestinal tract. A typical reactor setup for SHIME consists of three consecutive reactors; the first reactor simulates the different steps of food intake and digestion (representing the stomach and small intestine). The other two reactors simulate the large intestine (proximal and distal colon) and are inoculated with fecal samples from healthy donors. The experiment was divided into three different phases: a control phase (2 weeks, during which the reactors and fecal samples were stabilized to serve as a baseline for microbial community and activity), a treatment phase (3 weeks, during which CLX115 was added three times daily with food to simulate repeated ingestion and reveal the effects of oligomers), and a washout phase (2 weeks, to test the persistence of the effect after discontinuation of treatment). Samples were collected at different time points during the experiment and analyzed to determine the levels of SCFAs. SCFA (acetate, propionate, and butyrate) and BCFA (isobutyrate, isovalerate, and isocaproate) were determined by gas chromatography (GC2014-AOC 20i autosampler, Shimadzu Europa GmbH) as recently published (Ghyselinck, Verstrepen et al. 2020).

[0281] The results showed that CLX115 had a strong butyrate-producing effect during treatment ( Figure 4A (proximal colon) and Figure 4B (distal colon)) and strong propionic acid-enhancing effects ( Figure 5A (proximal colon) and Figure 5B No significant effects were observed on acetate and BCFA. After cessation of treatment, concentrations of these SCFAs generally returned to their pre-treatment levels. The ability of CLX115 to increase SCFA production suggests that this oligosaccharide may serve as a stimulator of GLP-1 production, thereby regulating blood glucose levels.

[0282] Gut microbial fermentative activity was determined by measuring gas production in the presence of CLX115. Triplicate short-term colonic simulations were performed using inocula collected from proximal colonic vessels at the end of the treatment phase during the SHIME experiment. We compared this with FOS treatment and quantified total gas production. A negative control, which received no treatment, was included. The negative control was inoculated with a sample from the control phase.

[0283] Gas formation was measured using a manometer with a needle attached (Handheld Pressure Indicator CPH6200; Wika, Echt, The Netherlands). The results showed that CLX115 produced less gas (% added) than FOS ( Figure 6 This data suggests that CLX115 in vitro total gas production in this system is greater than 100 kPa, which is associated with bloating. Therefore, CLX115 is a useful agent for glucose control that has a lower risk of bloating than the common prebiotic FOS.

[0284] Example 4: Optimization of hydrogen peroxide and copper (II) sulfate concentrations in copper-Fenton depolymerization of β-glucan.

[0285] The use of iron- and copper-based Fenton depolymerization has been demonstrated in several previous publications (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, copper-based Fenton depolymerization has never been fully optimized. Here, we demonstrate the effect of hydrogen peroxide concentration on copper-based Fenton depolymerization of β-glucans, specifically β-glucans from cereals.

[0286] 4 g of β-glucan was weighed out in three vials and dissolved to a concentration of 10% in 50 mM sodium acetate buffer, pH 5.5, containing 2%, 3%, or 4% hydrogen peroxide. The mixture was heated to 55°C in a shaking incubator. The reaction was initiated by adding copper (II) sulfate to a final concentration of 0.25 mM, 0.50 mM, or 1.00 mM and incubated at 55°C for 2 hours. After 2 hours, the mixture was removed from the incubator and cooled to below 15°C. Next, concentrated ammonium hydroxide was added to a final concentration of 0.39 M and incubated at 45°C in a shaking incubator for 2 hours. The reaction was stopped by freezing at -80°C and lyophilizing to dryness. The lyophilized material was rehydrated with a minimum of ultrapure H2O and then diluted with 200% food-grade ethanol to a 60% ethanol solution. The suspension was extracted by centrifugation at 4700 rpm for 15 minutes at -10°C, and the supernatant was decanted. The subsequent supernatant volume was reduced by rotary evaporation and lyophilized to yield a crystalline solid.

[0287] It was concluded that the reaction containing a 4% H₂O₂ concentration produced the optimal oligosaccharide molecular weight and the highest yield. This achieved increased yield while maintaining low free monosaccharide production and high purity. Furthermore, the reaction required a copper(II) sulfate concentration of 0.50 mM to 1.00 mM to provide the optimal oligosaccharide molecular weight and highest yield. Therefore, it can be concluded that hydrogen peroxide and copper(II) sulfate concentrations can be used to optimize the yield and / or molecular weight distribution of β-glucan oligosaccharides.

[0288] Example 5: Optimization of ammonium hydroxide concentration in copper-Fenton depolymerization of β-glucan.

[0289] The use of iron- and copper-based Fenton depolymerization has been demonstrated in several previous publications (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, copper-based Fenton depolymerization has never been fully optimized. Here, we demonstrate the effect of copper concentration on copper-based Fenton depolymerization of β-glucans, specifically β-glucans from cereals.

[0290] A beaker containing 50 mM ammonium acetate buffer at a pH of 5.50 was heated to 55°C. Next, the β-glucan was slowly stirred to a final concentration of 10%, and then hydrogen peroxide was added to a concentration of 4%. When the mixture reached 55°C, the reaction was started by adding copper (II) sulfate to a concentration of 0.75 mM. The reaction was stirred at 55°C for 2 hours and then immediately cooled to below 15°C. To begin the alkaline lysis step, concentrated ammonium hydroxide was added to a final concentration of 0.58 M or 0.76 M. The reaction was carried out at 45°C for 2 hours and then stopped and vacuum filtered using a GD120 filter and a Buchner funnel. The filtrate was then treated with MB10 resin (10% w / v) to a conductivity of approximately 90 μS / cm. The resin was removed by vacuum filtration through a glass frit funnel and then freezing and lyophilizing the filtrate to dryness. The lyophilized material was rehydrated with minimal ultrapure H2O and then diluted with 200°C food-grade ethanol to a 60% ethanol solution. The suspension was extracted by centrifugation at 4700 rpm for 15 minutes at -10°C, followed by decanting the supernatant. The subsequent supernatant volume was reduced by rotary evaporation and lyophilization to produce a white, fluffy, crystalline solid.

[0291] It was concluded that reactions containing ammonium hydroxide at concentrations between 0.58 M and 0.76 M produced near-optimal oligosaccharide molecular weights and the highest yields. This achieved increased yield while maintaining low free monosaccharide production and high purity. Therefore, it can be concluded that ammonium hydroxide concentration can be used to optimize the yield and / or molecular weight distribution of β-glucan oligosaccharides.

[0292] Example 6: Optimized conditions for copper-Fenton depolymerization of β-glucan.

[0293] The use of iron- and copper-based Fenton depolymerization has been demonstrated in several previous publications (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, copper-based Fenton depolymerization has never been fully optimized. Here, we present a set of optimized parameters and provide guidance for copper-based Fenton depolymerization of β-glucans, particularly those from cereals.

[0294] A solution containing 4% hydrogen peroxide and 43.4 mM ammonium acetate buffer at a pH of 5.5 is heated to 55°C. β-glucan (or other) polysaccharide is gradually stirred to a final concentration of 10%. To begin the first reaction step, copper (II) sulfate is added to a final concentration of 0.75 mM. The reaction is carried out at 55°C for 2 hours and then cooled to below 15°C. Next, to begin the second reaction step, concentrated ammonium hydroxide is added to a final concentration of 0.67 M. The reaction is stirred at 45°C for 2 hours. The reaction is vacuum filtered using a GD120 filter and a Buchner funnel, and then treated with MB10 resin (10% w / v) until the conductivity is below the threshold of 100 μS / cm. The resin is removed by vacuum filtration using a glass fritted funnel, the filtrate is frozen, and then lyophilized to dryness. The lyophilized product mixture is then dissolved in a minimum of ultrapure H2O, and a certain volume of 200-degree food-grade ethanol is added to produce a 60% ethanol solution. The solution was then separated by centrifugation (4700 RPM, 15 min, -10°C). The supernatant was carried on to the next step, while the precipitate was redissolved with minimal ultrapure H2O, followed by the addition of a volume of 200°C food-grade ethanol to produce a 60% ethanol solution. The solution was then separated by centrifugation (4700 RPM, 15 min, -10°C). The accumulated supernatant volume was reduced by rotary evaporation and then lyophilized to produce a fluffy white crystalline solid.

[0295] Upon analysis, the depolymerization product showed a composition in which, as measured by hydrolyzed monosaccharide composition analysis, approximately 87.5% by mass comprised glucose and approximately 4.5% by mass comprised arabinose. The glycosidic bond composition approximately comprised the amounts listed in Table B for CLX 115Cu. The oligosaccharide composition approximately comprised the values listed in Table J (CLX115Cu) as measured by oligosaccharide analysis. The molecular weight distribution of the composition approximately comprised the values listed in Table K (CLX115Cu) as measured by refractive index detection (RID) (see also Figure 7B ).

[0296] Therefore, it can be concluded that the optimized conditions can produce the desired oligosaccharide molecular weight distribution and yield.

[0297] Example 7: Optimized conditions for iron-based Fenton depolymerization of β-glucan.

[0298] The use of iron- and copper-based Fenton depolymerization has been demonstrated in several previous publications (WO2021097138A1, WO2018236917A1, WO2020247389A1). Here, we present a set of optimized parameters for iron-based Fenton depolymerization of β-glucans, particularly β-glucans from cereals.

[0299] A solution containing 7% hydrogen peroxide and 43.4 mM ammonium acetate buffer at a pH of 5.5 is heated to 55°C. β-glucan (or other) polysaccharide is gradually stirred to a final concentration of 5%. To begin the first reaction step, iron (II) sulfate is added to a final concentration of 1.15 mM. The reaction is carried out at 55°C for 2 hours and then cooled to below 15°C. Next, to begin the second reaction step, concentrated ammonium hydroxide is added to a final concentration of 0.39 M. The reaction is stirred at 45°C for 2 hours. The reaction is vacuum filtered using a GD120 filter and a Buchner funnel, and then treated with MB10 resin (10% w / v) until the conductivity is below the threshold of 100 μS / cm. The resin is removed by vacuum filtration using a glass fritted funnel, the filtrate is frozen, and then lyophilized to dryness. The lyophilized product mixture is then dissolved in a minimum of ultrapure H2O, and a certain volume of 200-degree food-grade ethanol is added to produce a 60% ethanol solution. The solution was then separated by centrifugation (4700 RPM, 15 min, -10°C). The supernatant was carried on to the next step, while the precipitate was redissolved with minimal ultrapure H2O, followed by the addition of a volume of 200°C food-grade ethanol to produce a 60% ethanol solution. The solution was then separated by centrifugation (4700 RPM, 15 min, -10°C). The accumulated supernatant volume was reduced by rotary evaporation and then lyophilized to produce a fluffy white crystalline solid.

[0300] Upon analysis, the depolymerization product showed a composition wherein approximately 95% by mass comprised glucose and approximately 2% by mass comprised arabinose, as measured by hydrolyzed monosaccharide composition analysis. The glycosidic bond composition approximately comprised the amounts listed in Table B for CLX 115. The composition approximately comprised the 1H-13C HSQC NMR correlations for CLX 115 listed in Table A. The oligosaccharide composition approximately comprised the values listed in Table G (CLX 115) as measured by oligosaccharide analysis. The molecular weight distribution of the composition approximately comprised the values listed in Table K (CLX 115) as measured by refractive index detection (RID) (see also Figure 7C The composition has a dynamic viscosity of about 1.382 mPa*s at 100 mg / mL at 25°C.

[0301] Therefore, it can be concluded that the optimized conditions can produce the desired oligosaccharide molecular weight distribution and yield.

[0302] Example 8. Comparison of the depolymerization of β-glucan by copper-based Fenton and iron-based Fenton.

[0303] The use of iron- and copper-based Fenton depolymerization has been demonstrated in several previous publications (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, copper-based Fenton depolymerization has never been fully optimized. Here, we demonstrate a comparison of copper- and iron-based Fenton depolymerization of β-glucans, particularly those from cereals.

[0304] Copper-based Fenton depolymerization was performed as described in Example 6. Iron-based Fenton depolymerization was performed as described in Example 7.

[0305] in conclusion Therefore, it can be concluded that both copper- and iron-Fenton depolymerization of β-glucan can produce similar oligosaccharide profiles. However, copper-Fenton depolymerization is considered to be more economically feasible than iron-Fenton depolymerization due to its higher substrate loading capacity, lower metal catalyst concentration, and lower hydrogen peroxide concentration.

[0306] Example 9: Capsule composition

[0307] Capsules were prepared by filling approximately 0.75 g of β-glucan oligosaccharide into 000 gelatin capsules using a filling machine. The capsules were then sealed. β-glucan oligosaccharide was in the form of a free-flowing powder.

[0308] Example 10: Nutritional composition

[0309] β-glucan oligosaccharide was added to a rotary blender at a mass ratio of 4:1. 0.25 wt% silicon dioxide was added to the blender and the mixture was blended for 10 minutes. The mixture was then agglomerated in a fluidized bed, filled into 5 g stick packs, and the packs were sealed.

[0310] Example 11: In vivo evaluation of the effect of β-glucan on blood glucose peaks during an oral maltose challenge.

[0311] Some long carbohydrate structures have been described to significantly promote blood glucose regulation in normal mice (Cao et al. 2016). In vivo assays were performed using mice to evaluate the effect of CLX115-Cu on glucose spikes after administration of maltose alone as a control or mixed with an equal amount of CLX115-Cu.

[0312] CLX115-Cu was produced using isolated β-glucan according to the method described in Example 8. Maltose or maltose plus CLX115-Cu was administered to two groups of healthy C57BL / 6 male mice (n = 15 mice / group). Mice received a single oral dose of 200 mg / mL of maltose alone or mixed with an equal amount of CLX115-Cu. Following administration, blood glucose levels were measured using a handheld glucometer before dosing (0 h), and 0.25 h, 0.5 h, 1 h, 1.5 h, and 2 h after dosing.

[0313] Blood glucose levels showed the expected trend during the experiment, with glucose peaking between 30 and 90 minutes, followed by a drop in blood glucose between 90 and 120 minutes ( Figure 9A ). CLX115-Cu significantly reduced peak blood glucose levels relative to control (maltose alone) (two-way ANOVA) ( Figure 9B This result suggests that CLX115-Cu may interact with α-glucosidase, thereby reducing the amount of glucose in the blood, as described in Example 2.

[0314] Example 12: Use of copper-based Fenton depolymerization using sodium hydroxide as an alternative base.

[0315] The use of optimized copper-based Fenton depolymerization was demonstrated in the previous examples of this application. Earlier applications demonstrated the use of sodium hydroxide as an alternative base (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, the optimized copper-based Fenton depolymerization described here has not been described with an alternative sodium hydroxide base. Here, we demonstrate a set of optimized parameters and provide guidance for copper-based Fenton depolymerization of β-glucan using an alternative sodium hydroxide base.

[0316] A solution containing 4% hydrogen peroxide and 43.4 mM ammonium acetate buffer at pH 5.5 is heated to 55°C. β-glucan (or other) polysaccharide is gradually stirred to a final concentration of 10%. To begin the first reaction step, copper (II) sulfate is added to a final concentration of 0.75 mM. The reaction is carried out at 55°C for 2 hours and then cooled to below 15°C. Next, to begin the second reaction step, 50% w / v sodium hydroxide solution is added to a final pH of 9; alternatively, the pH can be adjusted to any value in the range of 8 to 12, and varying degrees of depolymerization can still be achieved. The reaction is stirred at 45°C for 2 hours. The reaction is vacuum filtered using a GD120 filter and a Buchner funnel, and then treated with MB10 resin (10% w / v) until the conductivity is below the threshold of 100 μS / cm. The resin is removed by vacuum filtration using a glass fritted funnel, and the filtrate is frozen and then lyophilized to dryness. The lyophilized product mixture is then dissolved in a minimum of ultrapure H2O, followed by the addition of a volume of 200-degree food-grade ethanol to produce a 60% ethanolic solution. The solution is then separated by centrifugation (4700 RPM, 15 min, -10°C). The supernatant is continued to the next step while the precipitate is dissolved again with a minimum of ultrapure H2O, followed by the addition of a volume of 200-degree food-grade ethanol to produce a 60% ethanolic solution. The solution is then separated by centrifugation (4700 RPM, 15 min, -10°C). The accumulated supernatant volume is reduced by rotary evaporation and then lyophilized to produce a fluffy white crystalline solid.

[0317] It is expected that copper-based Fenton depolymerization paired with sodium hydroxide base will yield a depolymerization product that is the same or substantially similar to that described in Example 6.

[0318] Example 13: Use of iron-based Fenton depolymerization with sodium hydroxide as an alternative base.

[0319] The use of optimized iron-based Fenton depolymerization was demonstrated in the previous examples of this application. Earlier applications demonstrated the use of sodium hydroxide as an alternative base (WO2021097138A1, WO2018236917A1, WO2020247389A1). However, the optimized iron-based Fenton depolymerization described here has not been described with an alternative sodium hydroxide base. Here, we demonstrate a set of optimized parameters and provide guidance for iron-based Fenton depolymerization of β-glucan using an alternative sodium hydroxide base.

[0320] A solution containing 4% hydrogen peroxide and 43.4 mM ammonium acetate buffer at pH 5.5 is heated to 55°C. β-glucan (or other) polysaccharide is gradually stirred to a final concentration of 10%. To begin the first reaction step, iron (II) sulfate is added to a final concentration of 1.15 mM. The reaction is carried out at 55°C for 2 hours and then cooled to below 15°C. Next, to begin the second reaction step, 50% w / v sodium hydroxide solution is added to a final pH of 9; alternatively, the pH can be adjusted to any value in the range of 8 to 12, and varying degrees of depolymerization can still be achieved. The reaction is stirred at 45°C for 2 hours. The reaction is vacuum filtered using a GD120 filter and a Buchner funnel, and then treated with MB10 resin (10% w / v) until the conductivity is below the threshold of 100 μS / cm. The resin is removed by vacuum filtration using a glass fritted funnel, and the filtrate is frozen and then lyophilized to dryness. The lyophilized product mixture is then dissolved in a minimum of ultrapure H2O, followed by the addition of a volume of 200-degree food-grade ethanol to produce a 60% ethanolic solution. The solution is then separated by centrifugation (4700 RPM, 15 min, -10°C). The supernatant is continued to the next step while the precipitate is dissolved again with a minimum of ultrapure H2O, followed by the addition of a volume of 200-degree food-grade ethanol to produce a 60% ethanolic solution. The solution is then separated by centrifugation (4700 RPM, 15 min, -10°C). The accumulated supernatant volume is reduced by rotary evaporation and then lyophilized to produce a fluffy white crystalline solid.

[0321] It is expected that the iron-based Fenton depolymerization paired with sodium hydroxide base will give a depolymerization product that is the same or substantially similar to that described in Example 7.

[0322] References

[0323] All references listed below or elsewhere in this specification are incorporated herein by reference in their entirety for all purposes:

[0324] Cao, Y., Zoe, S., Xu, H., Li, M., Tong, Z., Xu, M., Xu,

[0325] Benalla, W., Bellahcen, S., Bnouham, M. Antidiabetic medicinal plants assource of alpha glucosidase inhibitors. Current Diabetes Rev. 2010, 6: 247-254

[0326] Everard.A., Cani, PDGut microbiota and GLP-1.Rev Endocr MEtab Disord, 2014, 15: 189-196

[0327] Aspects of the Invention

[0328] Multiple aspects are contemplated herein, several of which are listed in the following paragraphs. It is expressly contemplated that any aspect or portion thereof may be combined to form an aspect. Furthermore, it is expressly contemplated that any aspect (e.g., A13 aspect) that references an aspect (e.g., A1 aspect) with sub-aspects (e.g., A1a, A1b, A1c, etc.) having the same top-level number (e.g., A1a, A1b, A1c, etc.) necessarily includes reference to those A1a, A1b, A1c, etc. sub-aspects. In other words, if A13 aspect references A1 aspect, and there are A1a and A1b aspects, then A13 aspect also references A1a or A1b. Furthermore, although the aspects below are subdivided into A, B, C, D, etc. aspects, it is expressly contemplated that the aspects in each subdivision A, B, C, D, etc. may be combined in any manner. In addition, the term "any of the foregoing aspects" means any aspect that appears before the aspect containing the phrase (in other words, the sentence "Aspect B13: any of Aspects B1-B12 or the method of any of the foregoing aspects, ..." means that any aspect before Aspect B13 is referenced, including Aspects B1-B12 and all "A" aspects). For example, it is contemplated that optionally, any method or composition of any of the following aspects can be used or combined with any other aspect provided below. In addition, for example, it is contemplated that any embodiment described elsewhere herein (including above in this paragraph) can be optionally combined with any aspect listed below. In some examples in the aspects below or elsewhere herein, two open ranges are disclosed as being combinable into one range. For example, "at least X" is disclosed as being combinable with "less than Y" to form a range, where X and Y are numerical values. For the purpose of forming ranges herein, it is expressly contemplated that "at least X" is combined with "less than Y" to form an XY range that includes value X and value Y.

[0329] Aspect A1. A method for treating a glucose-related metabolic disorder in a subject, the method comprising enterally administering to the subject an effective amount of β-glucan oligosaccharide before and / or during ingestion of a glucose source by the subject.

[0330] Aspect A2: The method of Aspect A1, wherein the glucose-related metabolic disorder is diabetes mellitus.

[0331] Aspect A2a: The method of Aspect A1 or A2, wherein the glucose-related metabolic disorder is type 1 diabetes.

[0332] Aspect A2b: The method of any of Aspects A1-A2a, wherein the glucose-related metabolic disorder is type 2 diabetes.

[0333] Aspect A2c: The method of any of Aspects A1-A2b, wherein the glucose-related metabolic disorder is gestational diabetes.

[0334] Aspect A3: The method of any of Aspects A1-A2c, wherein the glucose-related metabolic disorder is metabolic syndrome.

[0335] Aspect B1: A method for attenuating a postprandial glucose response in a subject, the method comprising enterally administering an effective amount of β-glucan oligosaccharide to the subject before and / or during the subject's ingestion of a glucose source.

[0336] Aspect B2: The method according to Aspect B1 or any of the preceding aspects, wherein the subject is at risk for diabetes, is overweight and / or obese, is pregnant and / or suffers from diabetes.

[0337] Aspect C1: A method for reducing the risk of a prediabetic and / or obese subject developing type 2 diabetes, the method comprising attenuating the subject's postprandial glucose response by intraenterically administering an effective amount of β-glucan oligosaccharides to the subject before and / or during the subject's intake of a glucose source.

[0338] Aspect D1: A method for reducing the HbA1c level of a subject, the method comprising attenuating the subject's postprandial glucose response for at least 2 months by enterally administering an effective amount of β-glucan oligosaccharide to the subject before and / or during the subject's ingestion of a glucose source.

[0339] Aspect E1: The method of any one of Aspects A1-D1, wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.5g to about 20g, for example, from about 0.5g to about 20g, from about 0.5g to about 15g, from about 0.5g to about 10g, from about 0.5g to about 7.5g, from about 0.75g to about 20g, from about 0.75g to about 15g, from about 0.75g to about 15g, from about 0.75g to about 10g, from about 0.75g to about 7.5g, from about 1g to about 20g, from about 1g to about 15g, from about 1g to about 10g, or from about 1g to about 7.5g.

[0340] Aspect E2: The method of Aspects A1-D1 or any of the preceding aspects, wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.75g to about 7.5g, for example, from about 0.75g to about 20g, from about 0.75g to about 15g, from about 0.75g to about 15g, from about 0.75g to about 10g, from about 0.75g to about 7.5g, from about 1g to about 20g, from about 1g to about 15g, from about 1g to about 10g, or from about 1g to about 7.5g.

[0341] Aspect E3: The method of any of Aspects A1-E2, wherein the β-glucan oligosaccharide is administered at most 2 hours, at most 1.5 hours, at most 1 hour, or at most 30 minutes prior to ingestion of the glucose source by the subject.

[0342] Aspect E4: The method of any of Aspects A1-E3, wherein the β-glucan oligosaccharide is administered at most 30 minutes, at most 20 minutes, or at most 15 minutes prior to ingestion of the glucose source by the subject.

[0343] Aspect E5: The method of Aspects A1-D1 or any of the preceding aspects, wherein the β-glucan oligosaccharide is administered up to 30 minutes, up to 20 minutes, or up to 15 minutes prior to the subject's ingestion of the glucose source, and wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.5 g to about 20 g (e.g., about 0.5 g to about 20 g, about 0.5 g to about 15 g, about 0.5 g to about 10 g, about 0.5 g to about 7.5 g, about 0.75 g to about 20 g, about 0.75 g to about 15 ... to about 10 g, about 0.75 g to about 7.5 g, about 1 g to about 20 g, about 1 g to about 15 g, about 1 g to about 10 g, or about 1 g to about 7.5 g) or optionally in a range of about 0.75 g to about 7.5 g (e.g., about 0.75 g to about 7.5 g, for example, about 0.75 g to about 20 g, about 0.75 g to about 15 g, about 0.75 g to about 15 g, about 0.75 g to about 10 g, about 0.75 g to about 7.5 g, about 1 g to about 20 g, about 1 g to about 15 g, about 1 g to about 10 g, or about 1 g to about 7.5 g).

[0344] Aspect E6: The method of any of Aspects A1-E5, wherein the β-glucan oligosaccharide inhibits salivary amylase or pancreatic amylase.

[0345] Aspect E7: The method of any of Aspects A1-E6, wherein the β-glucan oligosaccharide inhibits the SGLT1 glucose transporter.

[0346] Aspect E8: The method of any one of Aspects A1-E7, wherein the β-glucan oligosaccharide inhibits α-glucosidase.

[0347] Aspect E9: The method of any one of Aspects A1-E8, wherein the β-glucan oligosaccharide comprises β-1,3 linked glucose residues and β-1,4 linked glucose residues.

[0348] Aspect E9a: The method of any one of Aspects A1-E9, wherein the β-glucan oligosaccharide comprises CLX101, CLX102, CLX112, CLX115, CLX115Cu, CLX123, CLX125, or a combination thereof.

[0349] Aspect E9b: The method of any one of Aspects A1-E9a, wherein the β-glucan oligosaccharide comprises CLX112, CLX115, CLX115Cu, or a combination thereof.

[0350] Aspect E10: The method of any one of Aspects A1-E9, wherein the β-glucan oligosaccharide comprises a ratio of β-1,3 linked glucose residues:β-1,4 linked glucose residues of 1:1 to 1:5 (e.g., 1:1 to 1:5, 1:2 to 1:5, 1:3 to 1:5, or 1:1 to 1:4).

[0351] Aspect E11: The method of any one of Aspects A1-E10, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 10,000 Da (eg, 100 Da to 10,000 Da, 500 Da to 10,000 Da, or 1,000 Da to 10,000 Da).

[0352] Aspect E12: The method of any of Aspects A1-E11, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 8,000 Da (eg, 100 Da to 8,000 Da, 500 Da to 8,000 Da, or 1,000 Da to 8,000 Da).

[0353] Aspect E13: The method of any of Aspects A1-E12, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 7,500 Da (eg, 100 Da to 7,500 Da, 500 Da to 7,500 Da, or 1,000 Da to 7,5000 Da).

[0354] Aspect E14: The method of any of Aspects A1-E13, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 5,000 Da (eg, 100 Da to 5,000 Da, 500 Da to 5,000 Da, or 1,000 Da to 5,000 Da).

[0355] Aspect E15: The method of any one of Aspects A1-E14, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits (e.g., 3 to 30 subunits, 3 to 25 subunits, 5 to 30 subunits, 5 to 25 subunits, 10 to 30 subunits, or 10 to 25 subunits), wherein at least 50% of the subunits, at least 75% of the subunits, or 100% of the subunits are β-1,3 glucose residues, β-1,4 glucose residues, or a combination thereof.

[0356] Aspect E15a: The method of Aspect E15 or any of the preceding aspects, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits (e.g., 3 to 30 subunits, 3 to 25 subunits, 5 to 30 subunits, 5 to 25 subunits, 10 to 30 subunits, or 10 to 25 subunits), wherein each subunit is independently a β-1,3 glucose residue or a β-1,4 glucose residue.

[0357] Aspect E16: The method of any one of Aspects A1-E15a, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits (e.g., 3 to 30 subunits, 3 to 25 subunits, 5 to 30 subunits, 5 to 25 subunits, 10 to 30 subunits, or 10 to 25 subunits), wherein the β-glucan oligosaccharide comprises both β-1,3 glucose residues and β-1,4 glucose residues, and wherein at least 50% of the subunits, at least 75% of the subunits, or 100% of the subunits are β-1,3 glucose residues or β-1,4 glucose residues.

[0358] Aspect E16a: The method of any one of Aspects A1-E16, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits (e.g., 3 to 30 subunits, 3 to 25 subunits, 5 to 30 subunits, 5 to 25 subunits, 10 to 30 subunits, or 10 to 25 subunits), wherein the β-glucan oligosaccharide comprises both β-1,3 glucose residues and β-1,4 glucose residues, and wherein each subunit is independently a β-1,3 glucose residue or a β-1,4 glucose residue.

[0359] Aspect E17: The method of any one of Aspects A1 to E16a, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C (e.g., about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C, about 1 mPa*s to about 7.5 mPa*s at 100 mg / mL at 25°C, or about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C).

[0360] Aspect E18: The method of any one of Aspects A1-E16a or any of the preceding aspects, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C (e.g., about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C, or about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C, or about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C, or about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C).

[0361] Aspect E19: The method of any one of Aspects A1-E16a or any of the preceding aspects, wherein the β-glucan oligosaccharide has a dynamic viscosity of from about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C (e.g., from about 1 mPa*s to about 3 mPa*s at 100 mg / mL at 25°C, or from about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C, or from about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C).

[0362] Aspect E20: The method of any one of Aspects A1-E16a or any of the preceding aspects, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C (e.g., about 1 mPa*s to about 1.5 mPa*s at 100 mg / mL at 25°C or about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C).

[0363] Aspect E21: The method of any one of Aspects A1-E16a or any preceding aspect, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1.3 mPa*s to about 1.4 mPa*s at 100 mg / mL at 25°C.

[0364] Aspect E22: The method of any one of Aspects A1 to E21, wherein at least 70% by mass (e.g., at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 100 kDa (e.g., less than 100 kDa, less than 75 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0365] Aspect E23: The method of any one of Aspects A1-E21 or any preceding aspect, wherein at least 60% by mass (e.g., at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 50 kDa (e.g., less than 50 kDa, less than 40 kDa, less than 30 kDa, less than 25 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0366] Aspect E24: The method of any one of Aspects A1-E21 or any preceding aspect, wherein at least 50% by mass (e.g., at least 50% by mass, at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 15 kDa (e.g., less than 15 kDa, less than 10 kDa, or less than 5 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0367] Aspect E25: The method of any one of Aspects A1-E21 or any preceding aspect, wherein at least 50% by mass (e.g., at least 50% by mass, at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 5 kDa (e.g., less than 5 kDa, less than 4 kDa, less than 2.5 kDa, or less than 1 kDa, optionally greater than 0.1 kDa).

[0368] Aspect E26: The method of any one of Aspects A1-E21 or any preceding aspect, wherein at least 25% by mass (e.g., at least 25% by mass, at least 35% by mass, at least 45% by mass, at least 50% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 1 kDa (e.g., less than 1 kDa or less than 0.75 kDa, optionally greater than 0.1 kDa).

[0369] Aspect E27: The method of any one of Aspects A1-E26, wherein the β-glucan oligosaccharide is produced by the following steps: reacting the polysaccharide in the reaction mixture with a Fenton reagent having a peroxide agent and a metal ion to provide a treated polysaccharide; and lysing the treated polysaccharide with an alkali to produce a mixture of polysaccharide lysate products and / or oligosaccharides having polysaccharide characteristics, wherein the mixture is a β-glucan oligosaccharide.

[0370] Aspect E28: The method of aspect E27 or any of the preceding aspects, wherein the Fenton reagent comprises hydrogen peroxide and one or more metal ions selected from the group consisting of transition metals Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II) and Co(II), alkaline earth metals Ca(II) and Mg(II), and lanthanide Ce(IV).

[0371] Aspect E29: The method of aspects E27 or E28 or any of the preceding aspects, wherein the base is one or more bases selected from the group consisting of ammonium hydroxide, ammonium bicarbonate, ammonia, urea, sodium amide, dimethylamine, trimethylamine, pyridine and N,N-diisopropylethylamine, sodium hydroxide, calcium hydroxide, potassium hydroxide, barium hydroxide, lithium hydroxide.

[0372] Aspect E30: The method of any one of Aspects E27-E29 or any preceding aspect, wherein the base is one or more bases selected from ammonium hydroxide and sodium hydroxide.

[0373] Aspect E31: The method of any one of Aspects E27-E29 or any preceding aspect, wherein the base is a nitrogen-based cleavage reagent.

[0374] Aspect E32: The method of Aspect E31 or any preceding aspect, wherein the nitrogen-based cleavage reagent is also a peroxide quenching reagent, and the initiation of polysaccharide cleavage is simultaneous or substantially simultaneous with the initiation of peroxide quenching.

[0375] Aspect E33: The method of Aspect E31 or any preceding aspect, wherein the nitrogen-based cleavage agent is not a peroxide quencher, and the method further comprises initiating peroxide quenching with an additional peroxide quencher.

[0376] Aspect E34: The method of any one of Aspects E27-E33 or any preceding aspect, wherein the metal ion is a copper ion.

[0377] Aspect E35: The method of any one of Aspects E27-E34 or any preceding aspect, wherein the copper ion is Cu(II).

[0378] Aspect E36: The method of Aspect E35 or any preceding aspect, wherein Cu(II) is used in the reaction mixture at a concentration of about 0.25 mM to about 1.00 mM (e.g., about 0.25 mM to about 1.00 mM, about 0.25 mM to about 0.8 mM, about 0.5 mM to about 1 mM, or about 0.5 mM to about 0.8 mM).

[0379] Aspect E37: The method of Aspect E35 or any preceding aspect, wherein Cu(II) is used in the reaction mixture at a concentration of about 0.7 mM to about 0.8 mM.

[0380] Aspect E38: The method of Aspect E35 or any preceding aspect, wherein Cu(II) is used in the reaction mixture at a concentration of about 0.75 mM.

[0381] Aspect E39: The method of any one of Aspects E27-E38 or any preceding aspect, wherein the Fenton reagent comprises copper sulfate.

[0382] Aspect E40: The method of Aspect E39 or any preceding aspect, wherein copper sulfate is used in the reaction mixture at a concentration of about 0.25 mM to about 1.00 mM (e.g., about 0.25 mM to about 1.00 mM, about 0.25 mM to about 0.8 mM, about 0.5 mM to about 1 mM, or about 0.5 mM to about 0.8 mM).

[0383] Aspect E41: The method of Aspect E39 or any preceding aspect, wherein copper sulfate is used in the reaction mixture at a concentration of about 0.7 mM to about 0.8 mM.

[0384] Aspect E42: The method of Aspect E39 or any preceding aspect, wherein copper sulfate is used in the reaction mixture at a concentration of about 0.75 mM.

[0385] Aspect E43: The method of any one of Aspects E27-E42 or any of the preceding aspects, wherein the hydrogen peroxide concentration is from about 1% (v / v) to about 7% (v / v) (e.g., from about 1% (v / v) to about 7% (v / v), from about 1% (v / v) to about 6% (v / v), from about 1% (v / v) to about 5% (v / v), from about 1% (v / v) to about 4.5% (v / v), from about 3.5% (v / v) to about 7% (v / v), or from about 3.5% (v / v) to 6% (v / v)).

[0386] Aspect E44: The method of any one of Aspects E27-E42 or any preceding aspect, wherein the hydrogen peroxide concentration is about 3.5% (v / v) to 4.5% (v / v).

[0387] Aspect E45: The method of any one of Aspects E27-E42 or any preceding aspect, wherein the hydrogen peroxide concentration is about 4.0% (v / v).

[0388] Aspect E46: The method of any one of Aspects E27-E45 or any preceding aspect, wherein the base concentration is from about 0.2M to about 1M (e.g., from about 0.2M to about 1.00M, from about 0.2M to about 0.8M, from about 0.3M to about 1M, or from about 0.3M to about 0.8M).

[0389] Aspect E47: The method of any one of Aspects E27-E45 or any preceding aspect, wherein the base concentration is from about 0.3M to about 0.5M.

[0390] Aspect E48: The method of any one of Aspects E27-E45 or any preceding aspect, wherein the base concentration is about 0.4M.

[0391] Aspect E49: The method of any one of Aspects E27-E48 or any preceding aspect, wherein the base adjusts the pH of the mixture of polysaccharide cleavage products and / or oligosaccharides to a final pH of about 8 to 12.

[0392] Aspect E49a: The method of any one of Aspects E27-E48 or any preceding aspect, wherein the base adjusts the pH of the mixture of polysaccharide cleavage products and / or oligosaccharides to a final pH of about 8.5 to about 11.

[0393] Aspect E50: The method of any one of Aspects E27-E48 or any preceding aspect, wherein the base adjusts the pH of the mixture of polysaccharide cleavage products and / or oligosaccharides to a final pH of about 9.5 to about 10.5.

[0394] Aspect E51: The method of any one of Aspects E27-E48 or any preceding aspect, wherein the base adjusts the pH of the mixture of polysaccharide cleavage products and / or oligosaccharides to a final pH of about 10.

[0395] Aspect E52: The method of any one of Aspects E27-E51 or any preceding aspect, wherein the concentration of the polysaccharide reacted in the reaction mixture is about 2% (w / v) to about 20% (w / v) (e.g., about 2% (w / v) to 20% (w / v), about 2% (w / v) to 15% (w / v), about 2% (w / v) to 12% (w / v), about 5% (w / v) to 20% (w / v), about 5% (w / v) to 12% (w / v), or about 8% (w / v) to 20% (w / v)).

[0396] Aspect E53: The method of any one of Aspects E27-E51 or any preceding aspect, wherein the concentration of said polysaccharide reacted in said reaction mixture is from about 8% (w / v) to about 12% (w / v).

[0397] Aspect E54: The method of any one of Aspects E27-E51 or any preceding aspect, wherein the concentration of said polysaccharide reacted in said reaction mixture is about 10% (w / v).

[0398] Aspect E55: The method of any one of Aspects E27-E54 or any preceding aspect, wherein the at least one polysaccharide is derived from a cereal.

[0399] Aspect E55a: The method of any one of Aspects E27-E54 or any preceding aspect, wherein said polysaccharide is derived from a cereal.

[0400] Aspect E56: The method of any one of Aspects E27-E54 or any preceding aspect, wherein at least one polysaccharide is derived from oats or barley.

[0401] Aspect E56a: The method of any one of Aspects E27-E54 or any preceding aspect, wherein said polysaccharide is derived from oats or barley.

[0402] Aspect E57: The method of any one of Aspects E27-E56 or any preceding aspect, wherein at least one polysaccharide is a β-glucan polysaccharide.

[0403] Aspect E57a: The method of any one of Aspects E27-E57 or any preceding aspect, wherein said polysaccharide is a β-glucan polysaccharide.

[0404] Aspect E57b: The method of any one of Aspects E27-E57a or any preceding aspect, wherein the β-glucan polysaccharide is CLX112-PS, CLX115-PS, or a combination thereof.

[0405] Aspect E58: The method of Aspect E57 or E57b or any of the preceding aspects, wherein the β-glucan polysaccharide has a weight average molecular weight of 500 kDa or greater, optionally less than 10,000 kDa or less than 5,000 kDa.

[0406] Aspect F1: A β-glucan oligosaccharide, wherein at least 70% by mass (e.g., at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 100 kDa (e.g., less than 100 kDa, less than 75 kDa, or less than 50 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0407] Aspect G1: A β-glucan oligosaccharide, wherein at least 60% by mass (e.g., at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 50 kDa (e.g., less than 50 kDa, less than 40 kDa, less than 30 kDa, less than 25 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0408] Aspect H1: A β-glucan oligosaccharide, wherein at least 50% by mass (e.g., at least 50% by mass, at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharides have a molecular mass of less than 15 kDa (e.g., less than 15 kDa, less than 10 kDa, or less than 5 kDa, optionally greater than 0.1 kDa or 0.5 kDa).

[0409] Aspect II: A β-glucan oligosaccharide, wherein at least 50% by mass (e.g., at least 50% by mass, at least 60% by mass, at least 70% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharide has a molecular mass of less than 5 kDa (e.g., less than 5 kDa, less than 4 kDa, less than 2.5 kDa, or less than 1 kDa, optionally greater than 0.1 kDa).

[0410] Aspect J1: A β-glucan oligosaccharide, wherein at least 25% by mass (e.g., at least 25% by mass, at least 35% by mass, at least 45% by mass, at least 50% by mass, at least 75% by mass, at least 85% by mass, at least 95% by mass, or optionally 100% by mass) of the β-glucan oligosaccharide has a molecular mass of less than 1 kDa (e.g., less than 1 kDa or less than 0.75 kDa, optionally greater than 0.1 kDa).

[0411] Aspect J1a: The β-glucan oligosaccharide of any of Aspects F1-J1 or any preceding aspect, wherein the β-glucan oligosaccharide comprises CLX101, CLX102, CLX112, CLX115, CLX115Cu, CLX123, CLX125, or a combination thereof.

[0412] Aspect J1b: The β-glucan oligosaccharide of any of Aspects F1-J1a, wherein the β-glucan oligosaccharide comprises CLX112, CLX115, CLX115Cu, or a combination thereof.

[0413] Aspect K1: The β-glucan oligosaccharide of any one of aspects F1-J1 or any of the preceding aspects, wherein the β-glucan oligosaccharide is produced by the following steps: reacting a polysaccharide in a reaction mixture with a Fenton reagent having a peroxide agent and a metal ion to provide a treated polysaccharide; and lysing the treated polysaccharide with an alkali to produce a mixture of polysaccharide lysates and / or oligosaccharides having polysaccharide characteristics, wherein the mixture is a β-glucan oligosaccharide.

[0414] Aspect K2: The β-glucan oligosaccharide of aspect K1 or any of the preceding aspects, wherein the Fenton reagent comprises hydrogen peroxide and one or more metals selected from the group consisting of transition metals Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II) and Co(II), alkaline earth metals Ca(II) and Mg(II), and lanthanide Ce(IV).

[0415] Aspect K3: The β-glucan oligosaccharide of Aspect K1 or K2 or any of the preceding aspects, wherein the base is one or more bases selected from the group consisting of ammonium hydroxide, ammonium bicarbonate, ammonia, urea, sodium amide, dimethylamine, trimethylamine, pyridine and N,N-diisopropylethylamine, sodium hydroxide, calcium hydroxide, potassium hydroxide, barium hydroxide, lithium hydroxide.

[0416] Aspect K4: The β-glucan oligosaccharide of any of Aspects K1-K3 or any preceding aspect, wherein the base is one or more bases selected from ammonium hydroxide and sodium hydroxide.

[0417] Aspect K5: The β-glucan oligosaccharide of any of Aspects K1-K4 or any preceding aspect, wherein the base is a nitrogen-based cleavage reagent.

[0418] Aspect K6: The β-glucan oligosaccharide of Aspect K5 or any preceding aspect, wherein the nitrogen-based cleavage reagent is also a peroxide quenching reagent, and the initiation of polysaccharide cleavage is simultaneous or substantially simultaneous with the initiation of peroxide quenching.

[0419] Aspect K7: The β-glucan oligosaccharide of Aspect K5 or any preceding aspect, wherein the nitrogen-based cleavage agent is not a peroxide quencher, and the method further comprises initiating peroxide quenching with an additional peroxide quencher.

[0420] Aspect K8: The β-glucan oligosaccharide of any of Aspects K1-K7 or any preceding aspect, wherein the one or more metals are copper ions.

[0421] Aspect K9: The β-glucan oligosaccharide of any of Aspects K1-K8 or any of the preceding aspects, wherein the one or more metals is Cu(II).

[0422] Aspect K10: The β-glucan oligosaccharide of Aspect K9 or any of the preceding aspects, wherein Cu(II) is used at a concentration of about 0.25 mM to about 1.00 mM (e.g., about 0.25 mM to about 1.00 mM, about 0.25 mM to about 0.8 mM, about 0.5 mM to about 1 mM, or about 0.5 mM to about 0.8 mM).

[0423] Aspect K11: The β-glucan oligosaccharide of Aspect K9 or any preceding aspect, wherein Cu(II) is used at a concentration of about 0.7 mM to about 0.8 mM.

[0424] Aspect K12: The β-glucan oligosaccharide of Aspect K9 or any preceding aspect, wherein Cu(II) is used at a concentration of about 0.75 mM.

[0425] Aspect K13: The β-glucan oligosaccharide of any of Aspects K1-K12 or any of the preceding aspects, wherein the hydrogen peroxide concentration is from about 1% (v / v) to about 7% (v / v) (e.g., from about 1% (v / v) to about 7% (v / v), from about 1% (v / v) to about 6% (v / v), from about 1% (v / v) to about 5% (v / v), from about 1% (v / v) to about 4.5% (v / v), from about 3.5% (v / v) to about 7% (v / v), or from about 3.5% (v / v) to 6% (v / v)).

[0426] Aspect K14: The β-glucan oligosaccharide of any of Aspects K1-K12 or any preceding aspect, wherein the hydrogen peroxide concentration is from about 3.5% (v / v) to about 4.5% (v / v).

[0427] Aspect K15: The β-glucan oligosaccharide of any of Aspects K1-K12 or any preceding aspect, wherein the hydrogen peroxide concentration is about 4.0% (v / v).

[0428] Aspect K16: The β-glucan oligosaccharide of any of Aspects K1-K15 or any preceding aspect, wherein the base concentration is from about 0.2M to about 1M (e.g., from about 0.2M to about 1.00M, from about 0.2M to about 0.8M, from about 0.3M to about 1M, or from about 0.3M to about 0.8M).

[0429] Aspect K17: The β-glucan oligosaccharide of any of Aspects K1-K15 or any preceding aspect, wherein the base concentration is from about 0.3M to about 0.5M.

[0430] Aspect K18: The β-glucan oligosaccharide of any of Aspects K1-K15 or any preceding aspect, wherein the base concentration is about 0.4M.

[0431] Aspect K19: The β-glucan oligosaccharide of any one of Aspects K1 to K18 or any preceding aspect, wherein the concentration of the polysaccharide reacted in the reaction mixture is about 2% (w / v) to about 20% (w / v) (e.g., about 2% (w / v) to 20% (w / v), about 2% (w / v) to 15% (w / v), about 2% (w / v) to 12% (w / v), about 5% (w / v) to 20% (w / v), about 5% (w / v) to 12% (w / v), or about 8% (w / v) to 20% (w / v)).

[0432] Aspect K20: The β-glucan oligosaccharide of any of Aspects K1-K18 or any preceding aspect, wherein the concentration of the polysaccharide reacted in the reaction mixture is about 8% (w / v) to about 12% (w / v).

[0433] Aspect K21: The β-glucan oligosaccharide of any of Aspects K1-K18 or any preceding aspect, wherein the concentration of the polysaccharide reacted in the reaction mixture is about 10% (w / v).

[0434] Aspect K22: The β-glucan oligosaccharide of any of Aspects K1-K21 or any preceding aspect, wherein at least one polysaccharide is derived from a cereal.

[0435] Aspect K22a: The β-glucan oligosaccharide of any one of Aspects K1-K21 or any preceding aspect, wherein the polysaccharide is derived from a cereal.

[0436] Aspect K23: The β-glucan oligosaccharide of any of Aspects K1-K21 or any preceding aspect, wherein at least one polysaccharide is derived from oats or barley.

[0437] Aspect K23a: The β-glucan oligosaccharide of any one of Aspects K1-K21 or any preceding aspect, wherein the polysaccharide is derived from oats or barley.

[0438] Aspect K24: The β-glucan oligosaccharide of any one of Aspects K1-K21 or any preceding aspect, wherein at least one polysaccharide is a β-glucan polysaccharide.

[0439] Aspect K24a: The β-glucan oligosaccharide of any of Aspects K1-K21 or any of the preceding aspects, wherein at least one polysaccharide is CLX112-PS or CLX115-PS.

[0440] Aspect K24b: The β-glucan oligosaccharide of any one of Aspects K1-K21 or any preceding aspect, wherein the polysaccharide is a β-glucan polysaccharide.

[0441] Aspect K24c: The β-glucan oligosaccharide of any of Aspects K1-K21 or any preceding aspect, wherein the polysaccharide is CLX112-PS, CLX115-PS, or a combination thereof.

[0442] Aspect K25: The β-glucan oligosaccharide of Aspect K24 or K24a or any of the preceding aspects, wherein the β-glucan polysaccharide has a weight average molecular weight of 500 kDa or greater, optionally less than 10,000 kDa or less than 5,000 kDa.

[0443] Aspect L1: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any preceding aspect for treating a glucose-based metabolic disorder.

[0444] Aspect L2: Use of the β-glucan oligosaccharide of any one of aspects F1 to K25 or any of the preceding aspects for treating diabetes or metabolic syndrome.

[0445] Aspect L2a: The use of Aspect L2 or any of the preceding aspects for the treatment of type 2 diabetes.

[0446] Aspect L2b: Use of Aspect L2 or L2a or any of the preceding aspects for the treatment of type 1 diabetes.

[0447] Aspect L3: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects for attenuating a postprandial glucose response in a subject.

[0448] Aspect L3a: The use of Aspect L3 or any of the preceding aspects, wherein the subject is at risk for diabetes, is overweight and / or obese, is pregnant, and / or has diabetes.

[0449] Aspect L4: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects for reducing the risk of progression to type 2 diabetes in a prediabetic and / or obese subject.

[0450] Aspect L5: Use of the β-glucan oligosaccharide of any of Aspects F1-K25 or any of the preceding aspects for lowering HbA1c levels in a subject.

[0451] Aspect L6: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects for the preparation of a medicament for attenuating a postprandial glucose response in a subject.

[0452] Aspect L6a: The use of Aspect L6 or any of the preceding aspects, wherein the subject is at risk for diabetes, is overweight and / or obese, is pregnant, and / or has diabetes.

[0453] Aspect L7: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects for the preparation of a medicament for the treatment of a glucose-based metabolic disorder.

[0454] Aspect L8: The use of Aspect L7 or any of the preceding aspects for treating diabetes.

[0455] Aspect L8a: The use of Aspect L8 or any of the preceding aspects for treating type 1 diabetes.

[0456] Aspect L8b: Use of Aspect L8 or L8a or any of the preceding aspects for the treatment of type 2 diabetes.

[0457] Aspect L9: The use of Aspect L7 or any of the preceding aspects for the treatment of metabolic syndrome.

[0458] Aspect L10: Use of the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects for the preparation of a medicament for reducing the risk of progression to type 2 diabetes in a prediabetic subject and / or an obese subject.

[0459] Aspect L11: Use of a β-glucan oligosaccharide according to any one of aspects F1 to K25 or any of the preceding aspects for the preparation of a medicament for lowering HbA1c levels in a subject.

[0460] Aspect M1: A pharmaceutical composition comprising the β-glucan oligosaccharide of any one of aspects F1-K25 or any of the preceding aspects and a pharmaceutically acceptable carrier.

[0461] Aspect N1: A dietary supplement comprising the β-glucan oligosaccharide of any one of aspects F1-K25 or any preceding aspect and a food-grade carrier.

[0462] Statement Regarding Incorporation by Reference and Modifications

[0463] All references throughout this application, such as patent documents, including published or granted patents or equivalents; patent application publications; and non-patent literature documents or other source materials, to the extent that each reference is at least partially not inconsistent with the disclosure in this application, are hereby incorporated by reference in their entirety, as if individually incorporated by reference (e.g., partially contradictory references are incorporated by reference in their entirety except for the partially contradictory portions). All patents and publications mentioned in this specification are indicative of the technical level of those skilled in the art to which the invention pertains. The entire contents of the references cited herein are incorporated by reference to indicate the state of the art, in some cases as of their filing date, and if necessary, it is intended that this information can be used herein to exclude (e.g., not claim) specific embodiments in the prior art. For example, when claiming a compound, it should be understood that compounds known in the prior art, including certain compounds disclosed in the references disclosed herein (particularly in the cited patent documents), are not intended to be included in the claims.

[0464] The terms and expressions used herein are used as descriptive terms only and not as limiting terms, and when such terms and expressions are used, there is no intention to exclude any equivalents of the features shown and described or parts thereof, but it should be recognized that various modifications can be made within the scope of the invention claimed. Therefore, it should be understood that although the present invention has been specifically disclosed through preferred embodiments, exemplary embodiments and optional features, those skilled in the art may make modifications and changes to the concepts disclosed herein, and such modifications and changes are considered to be within the scope of the invention as defined in the appended claims. The specific embodiments provided herein are examples of useful embodiments of the present invention, and it will be apparent to those skilled in the art that a large number of variations of the devices, device components, and method steps set forth in this specification can be used to implement the present invention. It will be apparent to those skilled in the art that the methods and devices used for this method can include a large number of optional components and processing elements and steps.

[0465] As used herein and in the appended claims, the indefinite article ("a", "an") and the definite article ("the") in the singular include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art. Likewise, the terms indefinite article ("a" or "an"), "one or more" and "at least one" are used interchangeably herein. It should also be noted that the terms "comprising", "including" and "having" are used interchangeably. The expression "any of claims XX-YY (where XX and YY refer to the claim numbers)" is intended to provide multiple dependent claims in an alternative form and is, in some embodiments, interchangeable with the expression "any of claims XX-YY".

[0466] When a group of substituents is disclosed herein, it should be understood that all individual members of the group and all subgroups (including any isomers, enantiomers, and diastereomers of the group members) are disclosed separately. When Markush groups or other groupings are used herein, all individual members of the group and all possible combinations and subcombinations of the group are intended to be included in the disclosure separately. When a compound is described herein without specifying the specific isomers, enantiomers, and diastereomers of the compound, for example, in a chemical formula or chemical name, the description is intended to include each isomer and enantiomer of the compound described separately or any combination thereof. In addition, unless otherwise stated, all isotopic variants of the compounds disclosed herein are intended to be included in the disclosure. For example, it should be understood that any one or more hydrogens in the disclosed molecules can be replaced by deuterium or tritium. Isotopic variants of molecules can generally be used as standards for the determination of molecules and for chemical and biological studies related to molecules or their uses. Methods for preparing such isotopic variants are known in the art. Since it is known that those of ordinary skill in the art can name the same compound in different ways, the specific names of the compounds are intended to be exemplary.

[0467] Certain molecules disclosed herein may contain one or more ionizable groups [groups from which protons can be removed (e.g., -COOH) or protons can be added (e.g., amines) or can be quaternized (e.g., amines)]. All possible ionic forms of these molecules and their salts are intended to be individually included in the disclosure herein. With respect to the salts of the compounds herein, one of ordinary skill in the art can select from a variety of available counterions those suitable for preparing salts of the present invention for a given application. In a particular application, the choice of a given anion or cation for preparing a salt may result in an increase or decrease in the solubility of the salt.

[0468] Unless otherwise indicated, each device, system, formulation, combination of components, or method described or illustrated herein can be used to practice the present invention.

[0469] Whenever a range is given in the specification, such as a temperature range, time range, or composition range or concentration range, all intermediate ranges and subranges, as well as all individual values included in the given range, are intended to be included in the disclosure. It should be understood that any subrange or individual value within a range or subrange included in the description herein may be excluded from the claims herein.

[0470] All patents and publications mentioned in the specification are indicative of the state of the art of the art to which the invention pertains. The entire contents of the references cited herein are hereby incorporated by reference to indicate the state of the art as of their publication date or filing date, and it is intended that this information may be used herein to exclude specific embodiments in the prior art, if desired. For example, when claiming a composition of matter, it is understood that compounds known and available in the art prior to the applicant's invention, including compounds for which effective disclosure is provided in the references cited herein, are not intended to be included in the composition of matter claimed herein.

[0471] As used herein, "comprising" is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of excludes any element, step, or ingredient not specified in the claim elements. As used herein, "consisting essentially of does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim. For example, when referring to CLX or pharmaceutical compositions described herein, "consisting essentially of does not exclude other compounds, such as surfactants, flavorings, fillers, dyes, binders, buffers, preservatives, excipients, and carriers. In each instance herein, any one of the terms "comprising," "consisting essentially of," and "consisting of" may be replaced by either of the other two terms. The invention illustratively described herein may suitably be practiced in the absence of any element or elements, any limitation or limitations not specifically disclosed herein.

[0472] Those skilled in the art will understand that starting materials, biological materials, reagents, synthetic methods, purification methods, analytical methods, assay methods and biological methods other than those specifically illustrated can be used in the practice of the present invention without excessive experimentation. The present invention is intended to include all functional equivalents known in the art of any such materials and methods. The terms and expressions used are used as descriptive terms rather than restrictive terms, and when using these terms and expressions, there is no intention to exclude any equivalents of the features shown and described or parts thereof, but it should be recognized that various modifications can be made within the scope of the claimed invention. Therefore, it should be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, those skilled in the art can modify and change the concepts disclosed herein, and these modifications and changes are considered to be within the scope of the present invention as defined in the appended claims.

Claims

1. A method for treating a glucose-related metabolic disorder in a subject, the method comprising administering an effective amount of β-glucan oligosaccharide to the subject enterally before and / or during the subject's intake of a glucose source.

2. The method of claim 1, wherein the glucose-related metabolic disorder comprises diabetes mellitus, type 1 diabetes mellitus, type 2 diabetes mellitus, gestational diabetes, metabolic syndrome, or any combination thereof.

3. A method for attenuating a postprandial glucose response in a subject, the method comprising enterally administering to the subject an effective amount of β-glucan oligosaccharide before and / or during ingestion of a glucose source by the subject.

4. The method of claim 4, wherein the subject is at risk for diabetes, is overweight, obese, is pregnant, has diabetes, or any combination thereof.

5. A method for reducing the risk of a prediabetic and / or obese subject developing type 2 diabetes, the method comprising attenuating the subject's postprandial glucose response by enterally administering to the subject an effective amount of β-glucan oligosaccharides before and / or during the subject's ingestion of a glucose source.

6. A method for reducing HbA1c levels in a subject, the method comprising attenuating the subject's postprandial glucose response for at least 2 months by enterally administering to the subject an effective amount of β-glucan oligosaccharide before and / or during ingestion of a glucose source by the subject.

7. The method of any one of claims 1-6, wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.5 g to about 20 g.

8. The method of any one of claims 1-7, wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.75 g to about 15 g.

9. The method of any one of claims 1 to 8, wherein the β-glucan oligosaccharide is administered up to 2 hours prior to ingestion of a glucose source by the subject.

10. The method of any one of claims 1-9, wherein the β-glucan oligosaccharide is administered up to 30 minutes prior to ingestion of a glucose source by the subject.

11. The method of any one of claims 1-10, wherein the β-glucan oligosaccharide is administered up to 30 minutes prior to the subject's ingestion of a glucose source, and wherein the effective amount of the β-glucan oligosaccharide ranges from about 0.5 g to about 20 g or optionally ranges from about 0.75 g to about 15 g.

12. The method of any one of claims 1-11, wherein the β-glucan oligosaccharide inhibits salivary amylase or pancreatic amylase.

13. The method of any one of claims 1-12, wherein the β-glucan oligosaccharide inhibits the SGLT1 glucose transporter.

14. The method of any one of claims 1 to 13, wherein the β-glucan oligosaccharide inhibits α-glucosidase.

15. The method of any one of claims 1 to 14, wherein the β-glucan oligosaccharide comprises β-1,3 linked glucose residues and β-1,4 linked glucose residues.

16. The method of any one of claims 1 to 15, wherein the β-glucan oligosaccharide comprises a ratio of β-1,3 linked glucose residues to β-1,4 linked glucose residues of 1:1 to 1:

5.

17. The method of any one of claims 1 to 16, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 10,000 Da.

18. The method of any one of claims 1 to 17, wherein the β-glucan oligosaccharide has a weight average molecular weight (Mw) of less than 5,000 Da.

19. The method of any one of claims 1-18, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits, and wherein at least 50% of the subunits are β-1,3 glucose residues, β-1,4 glucose residues, or a combination thereof.

20. The method of any one of claims 1-19, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits, wherein the β-glucan oligosaccharide comprises both β-1,3 glucose residues and β-1,4 glucose residues, and wherein each subunit is independently a β-1,3 glucose residue or a β-1,4 glucose residue.

21. The method of any one of claims 1 to 20, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 10 mPa*s at 100 mg / mL at 25°C.

22. The method of any one of claims 1 to 20, wherein the β-glucan oligosaccharide has a dynamic viscosity of about 1 mPa*s to about 5 mPa*s at 100 mg / mL at 25°C.

23. The method of any one of claims 1 to 22, wherein at least 60% by mass of the β-glucan oligosaccharides have a molecular mass of less than 50 kDa.

24. The method of any one of claims 1 to 23, wherein at least 50% by mass of the β-glucan oligosaccharides have a molecular mass of less than 5 kDa.

25. The method of any one of claims 1-24 or 46-50, wherein the β-glucan oligosaccharide is produced by reacting a polysaccharide in a reaction mixture with a Fenton reagent having a peroxide agent and a metal ion to provide a treated polysaccharide; and lysing the treated polysaccharide with an alkali to produce a mixture of polysaccharide lysates and / or oligosaccharides having polysaccharide characteristics, the mixture being β-glucan oligosaccharide.

26. The method of claim 25, wherein the Fenton reagent comprises hydrogen peroxide and one or more metal ions selected from the group consisting of the transition metals Fe(II), Fe(III), Cu(I), Cu(II), Mn(II), Zn(II), Ni(II), and Co(II), the alkaline earth metals Ca(II) and Mg(II), and the lanthanide Ce(IV).

27. The process of claim 25 or 26, wherein the base is one or more bases selected from the group consisting of ammonium hydroxide, ammonium bicarbonate, ammonia, urea, sodium amide, dimethylamine, trimethylamine, pyridine and N,N-diisopropylethylamine, sodium hydroxide, calcium hydroxide, potassium hydroxide, barium hydroxide, lithium hydroxide.

28. The process of any one of claims 25 to 27, wherein the base is one or more bases selected from ammonium hydroxide and sodium hydroxide.

29. The method of any one of claims 25-27, wherein the base is a nitrogen-based cleavage reagent.

30. The method of claim 29, wherein the nitrogen-based cleavage reagent is also a peroxide quenching reagent, and the initiation of polysaccharide cleavage is simultaneous or substantially simultaneous with the initiation of peroxide quenching.

31. The method of claim 41, wherein The nitrogen-based cleavage agent is not a peroxide quencher, and the method further comprises initiating peroxide quenching with an additional peroxide quencher.

32. The method of any one of claims 26-31, wherein the one or more metal ions comprise copper ions.

33. The method of claim 32, wherein the copper ion is Cu(II).

34. The method of claim 33, wherein Cu(II) is used in the reaction mixture at a concentration of about 0.25 mM to about 1.00 mM.

35. The method of claim 33, wherein Cu(II) is used in the reaction mixture at a concentration of about 0.7 mM to about 0.8 mM.

36. The method of any one of claims 25-35, wherein the Fenton reagent comprises copper sulfate.

37. The method of claim 36, wherein copper sulfate is used in the reaction mixture at a concentration of about 0.25 mM to about 1.00 mM.

38. The method of any one of claims 25-37, wherein the hydrogen peroxide concentration is from about 1% (v / v) to about 7% (v / v).

39. The method of any one of claims 25-38, wherein the base concentration is from about 0.2 M to about 1 M.

40. The method of any one of claims 25 to 39, wherein the base adjusts the pH of the mixture of polysaccharide cleavage products and / or oligosaccharides to a final pH of about 8 to 12.

41. The method of any one of claims 25-40, wherein the polysaccharide is reacted in the reaction mixture at a concentration of about 2% (w / v) to about 20% (w / v).

42. The method of any one of claims 25 to 41, wherein at least one polysaccharide reacted in the reaction mixture is derived from a cereal.

43. The method of any one of claims 25 to 42, wherein at least one polysaccharide reacted in the reaction mixture is derived from oats or barley.

44. The method of any one of claims 25-43, wherein at least one polysaccharide reacted in the reaction mixture is a beta-glucan polysaccharide.

45. The method of claim 44, wherein the β-glucan polysaccharide has a weight average molecular weight of 500 kDa or greater.

46. A β-glucan oligosaccharide, wherein at least 70% by mass of the β-glucan oligosaccharides have a molecular mass of less than 100 kDa.

47. A β-glucan oligosaccharide, wherein at least 50% by mass of the β-glucan oligosaccharides have a molecular mass of less than 15 kDa.

48. The β-glucan oligosaccharide of claim 46 or 47, wherein the β-glucan oligosaccharide is derived from cereal β-glucan.

49. The β-glucan oligosaccharide of claim 48, wherein the β-glucan oligosaccharide is derived from oats or barley.

50. The β-glucan oligosaccharide of any one of claims 46-49, wherein the β-glucan oligosaccharide comprises 3 to 30 subunits, and wherein at least 50% of the subunits are β-1,3 glucose residues, β-1,4 glucose residues, or a combination thereof.

51. A pharmaceutical composition comprising the β-glucan oligosaccharide of any one of claims 46 to 50 and a pharmaceutically acceptable carrier.

52. A dietary supplement comprising the β-glucan oligosaccharide of any one of claims 46-50 and a food grade carrier.

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