Compositions for use in the treatment and / or prevention of type 2 diabetes, prediabetes, and / or symptoms thereof

Abstract

The present invention provides methods of using a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for the treatment and / or prevention of type 2 diabetes, prediabetes, and / or symptoms thereof in a subject. Additional uses of the composition are also provided herein.

Description

[Technical Field]

[0001] The present invention provides methods of using a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for the treatment and / or prevention of type 2 diabetes, prediabetes, and / or symptoms thereof in a subject. Additional uses of the composition are also provided herein. [Background technology]

[0002] There is increasing evidence that the gut microbiome is involved in the development of human diseases, including but not limited to disorders of the immune system, energy metabolism, lipid metabolism, and glucose metabolism [1, 2]. The proper development of metabolic functionality in the human body is strongly driven by the symbiotic relationship between this gut microbiome and the host organism. The microbial load in a single human is approximately 3.8 × 10 13 It is estimated that the total mass of the gut microbiome accounts for 200 g to 1 kg of an individual's total body weight [3-5]. A wide range of biological processes, including energy balance homeostasis and nutrient digestion, are influenced by the gut microbiome. The trigger for this function is the production of short-chain fatty acids (butyrate, propionate, and acetate), which are considered signaling molecules in tissues within and outside the gut [6]. Other functions include participation in the transport and metabolism of carbohydrates and amino acids, production of fat-soluble and water-soluble vitamins [7], elimination of xenobiotics and drugs, and bile acid metabolism [8]. Disruption of the composition and subsequent functionality of host-microbiome interactions can lead to chronic systemic inflammation, triggered by bacterial fragments appearing in circulating fluids as a result of gut barrier dysfunction. A dysfunctional gut barrier therefore has a significant triggering effect on host adiposity and insulin resistance [9].

[0003] Recent evidence suggests that certain bacterial species may interact with host metabolism, for example, through metabolite-mediated stimulation of gut hormones. Modulating the gut microbiome through exogenous supplementation with bacteria and probiotics may offer interesting opportunities for complementary treatment of several diseases

[10] . By definition, probiotics are substances contained in certain foods that can alter the composition of the gut microbiome, thereby affecting short-chain fatty acid production, satiety, weight control, energy homeostasis, and suppressing pathogen growth and immunomodulatory effects [11, 12]. Indeed, probiotics have been shown to be effective in a variety of clinical conditions, ranging from infant diarrhea, necrotizing enterocolitis, antibiotic-associated diarrhea, inflammatory bowel disease leading to cancer, female urinary tract infections, surgical infections, insulin resistance, and oxidative stress [13-15].

[0004] Diabetes mellitus (DM) is a chronic systemic disease with complications affecting most body organs. It is widespread, affecting approximately 9% of the world's population.

[21] Alterations in pancreatic beta cell function and insulin resistance result in relative insulin deficiency and impaired cellular response to insulin. A key symptom of the disease is hyperglycemia, which can lead to several serious secondary complications. [22,23] Antidiabetic medications are commonly used according to national and international treatment guidelines. [24,25] The most commonly used drug, metformin, may have limited efficacy and may have adverse effects on the composition of the intestinal bacterial microfauna.

[0005] Thus, there is a need for novel compositions, supplements, and / or therapeutic agents for use in the treatment and / or prevention of type 2 diabetes and / or its symptoms. Summary of the Invention [Problem to be solved by the invention]

[0006] Recently, it has been found that the gut microbiome is altered in diabetic patients, for example, by increasing the ratio of bacteroids to Clostridium species in the gut and by increasing the number of various opportunistic pathogens [26-29]. Circulating Gram-positive gut bacteria have been detected in blood samples from diabetic patients

[28] . Alterations in the gut microbiome can lead to metabolic endotoxemia due to lipopolysaccharide release, thus stimulating inflammation and insulin resistance

[30] . Enteroendocrine L cells in the epithelium play a role in causing inflammation, and their number has been positively or negatively correlated with the abundance of 25 bacterial taxa in the gut [31,32].

[0007] In light of these findings, we investigated the effects of probiotic supplementation on diabetes and diabetes-related complications.

[0008] The present inventors have previously developed a (fermented) rice bran-based composition (e.g., a nutritional supplement) containing Bacillus subtilis and Bacillus coagulans, which provides prebiotic and probiotic properties. The present inventors have now surprisingly shown that the composition (referred to herein as AB001) can be advantageously used to safely improve the cardiometabolic status of subjects, particularly those with type 2 diabetes and / or prediabetes. For example, the composition can advantageously provide a valuable supplement to existing concomitant treatments in subjects with type 2 diabetes and / or prediabetes.

[0009] The inventors have surprisingly shown that AB001, compared with placebo, has beneficial effects on glycemic control in patients with type 2 diabetes, as well as on biomarkers of inflammation (e.g., chronic systemic inflammation), insulin resistance, and beta-cell dysfunction. The inventors have therefore advantageously demonstrated that the composition described herein (AB001) can be used to stabilize glucose levels and / or improve glycemic control, particularly in subjects with type 2 diabetes and / or pre-diabetes. Advantageously, the compositions described herein can also be used to reduce insulin resistance, beta-cell dysfunction, and / or inflammation (e.g., chronic systemic inflammation and / or intestinal inflammation) in subjects with type 2 diabetes and / or pre-diabetes. In some examples, the inflammation can be intestinal and / or pancreatic inflammation.

[0010] In addition, the inventors surprisingly observed that changes in lipid profiles and other metabolic syndrome parameters in patients with type 2 diabetes were more favorable with AB001 than with placebo. Advantageously, the compositions described herein may therefore be used for the treatment and / or prevention of symptoms of metabolic syndrome in subjects with type 2 diabetes and / or prediabetes.

[0011] AB001 contains specially selected bacterial strains that preferably metabolize glucose and other carbohydrates to have a potent effect on glucose levels and insulin resistance. Sugars, such as glucose, metabolized from food and beverages in the intestinal tract serve, in part, as nutrients for cells of the microbiome (e.g., cells of the gut microbiome) before being reabsorbed into the blood as an energy reserve. Without being bound by theory, it is suggested that AB001 has four glucose-controlling effects: 1) the Bacilli strains in AB001 and their secreted enzymes act as additional glucose digestion factors (i.e., they act in addition to the subject's microbiome (e.g., gut microbiome) that was present prior to AB001 administration), further reducing glucose reabsorption into the blood; 2) AB001 reduces low-grade pancreatic inflammation, causing pancreatic beta cells to produce more insulin and less proinsulin; 3) AB001 increases cellular insulin sensitivity, resulting in a more normalized glucose-insulin mechanism; and 4) AB001 shifts the ratio of low-density lipids (LDL) to high-density lipids (HDL) in favor of higher concentrations of HDL. The fourth effect means that excess glucose is more highly converted to blood fats, such as triglycerides, which are then transported by HDL from the blood to the liver for degradation. In contrast, LDL does the opposite, transporting fat from the liver into the blood. Thus, in some cases, the action of AB001 is driven by the secretion of bioactive substances such as enzymes that break down complex organic molecules into nutrients, reactants, and energy. In some cases, (increased) glucose levels are normalized by the presence of more insulin.

[0012] It is suggested that, upon resuscitation in the intestinal tract, Bacillus spp. (and their endospores) of the compositions used according to the present invention begin to secrete a unique set of bioactive substances that scan the biochemical conditions of their microenvironment and optimize conditions, e.g., pH, conductivity, and electrolytes, for their survival and growth. Nutrients and substrates are essential for survival and subsequent growth. When carbohydrates (e.g., glucose, in particular) are present in the microenvironment, enzymes that target carbohydrates (e.g., glucose) are secreted to degrade the substrate. [Means for solving the problem]

[0013] Advantageously, the compositions used according to the present invention can be formulated as acid-resistant tablets or capsules. Such formulations are known to resist gastric acid and dissolve only upon reaching the duodenum. The Bacillus spp. of the compositions can then be released and settle in the upper intestinal tract, where they can remain for approximately one day before being excreted from the body through feces. The microbial species of the compositions described herein were selected to favorably and efficiently metabolize carbohydrates (especially glucose). One consequence of AB001 supplementation has been suggested to be a decrease in glucose absorption through the intestinal tract. Similar effects and corresponding biochemical findings have been observed when drugs belonging to the SGLT-1 inhibitor class (α-glucosidase inhibitors, e.g., acarbose or voglibose) are used in routine diabetes treatment. SGLT-1 inhibitors actively retain glucose molecules in the intestinal tract, where they are ultimately metabolized by resident bacteria in the large intestine, with methane gas as the end product. Therefore, patients treated with SGLT-1 inhibitors complained of significant problems with flatulence and abdominal bloating [39,40]. It is suggested that supplementation with AB001 means that less glucose reaches the blood.

[0014] Provided herein are compositions comprising one or more Bacillus species and rice bran for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject.

[0015] Also provided herein are compositions comprising one or more Bacillus species, rice bran, L-cysteine, and high molecular weight, low osmolality carbohydrates for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject.

[0016] Also provided herein are methods for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, the methods comprising administering to the subject a composition comprising one or more Bacillus species and rice bran.

[0017] Also provided herein are methods for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, the methods comprising administering to the subject a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

[0018] Suitably, the composition may be for the treatment and / or prevention of type 2 diabetes and / or symptoms thereof.

[0019] Preferably, administering the composition to a subject comprises: (a) capable of reducing one or more levels of HbA1c, glucose, insulin, intact proinsulin, and / or hsCRP in the blood; (b) may reduce the HOMA-IR score; and / or (c) It can increase blood adiponectin levels.

[0020] Preferably, administration of the composition to a subject can control blood glucose.

[0021] Preferably, administration of the composition to a subject is capable of degrading glucose.

[0022] Preferably, administration of the composition to a subject can metabolize glucose in the subject's digestive tract, optionally, administration of the composition to a subject can metabolize glucose in the subject's intestine, and further optionally, administration of the composition to a subject can metabolize glucose in the subject's small intestine.

[0023] Preferably, administration of the composition to a subject may reduce glucose absorption into the subject's blood.

[0024] Preferably, administration of the composition to a subject can reduce blood glucose levels in the subject.

[0025] Provided herein is the use of a composition comprising one or more Bacillus species and rice bran to degrade glucose.

[0026] (a) to reduce one or more levels of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, and / or hsCRP in the blood of a subject; (b) to reduce the HOMA-IR score in a subject; and / or (c) increasing blood adiponectin levels in a subject; Also provided herein are uses of compositions comprising one or more Bacillus species and rice bran.

[0027] Provided herein is the use of a composition comprising one or more Bacillus species and rice bran for controlling blood glucose in a subject.

[0028] Also provided herein is the use of a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate to degrade glucose.

[0029] (a) to reduce one or more levels of HbA1c, glucose, insulin, intact proinsulin, and / or hsCRP in the blood of a subject; (b) to reduce the HOMA-IR score in a subject; and / or (c) increasing blood adiponectin levels in a subject; Also provided herein are uses of compositions comprising one or more Bacillus species, rice bran, L-cysteine, and high molecular weight, low osmolality carbohydrates.

[0030] Further provided herein is a use of a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for controlling blood glucose in a subject.

[0031] Preferably, the one or more species of Bacillus may be selected from B. subtilis and B. coagulans.

[0032] Suitably, the composition may comprise B. subtilis and B. coagulans.

[0033] Preferably, a) the B. subtilis species may be selected from the group consisting of Bacillus subtilis strain DFM 0326 (LMG P-32899) and Bacillus subtilis strain DFM 1015 (LMG P-32900), and / or c) The B. coagulans species can be Bacillus coagulans strain DFM 0705 (LMG P-32921).

[0034] Preferably, the high molecular weight, low osmolality carbohydrate may be a dextrin.

[0035] Preferably, the composition may further comprise one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus. Preferably, the composition may further comprise one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genus Saccharomycetes (e.g., Brettanomyces bruxellensis (formerly Dekkerabruxellensis) and / or Millerozoma farinosa (formerly Pichia farinosa)).

[0036] Suitably, the composition may comprise at least about 20% w / w L-cysteine.

[0037] Preferably, the composition contains at least about 1 x 10 5 The bacteria may include bacteria of the genus Bacillus.

[0038] Suitably, the composition may comprise at least about 73% w / w rice bran.

[0039] Suitably, the composition may comprise at least about 0.5% w / w of a high molecular weight, low osmolality carbohydrate.

[0040] Suitably, the composition may further comprise one or more of vitamin B12, a magnesium salt of a fatty acid, calcium phosphate, potassium phosphate, silicon dioxide, and cellulose, and optionally, the magnesium salt of a fatty acid is magnesium stearate.

[0041] Suitably, the compositions may be formulated as acid-resistant tablets or capsules.

[0042] Suitably, the acid-resistant tablet or capsule may comprise a film coating, which comprises hydroxypropylmethylcellulose (HPMC).

[0043] Preferably, the one or more Bacillus species may be genetically unmodified.

[0044] Suitably, the composition may be for daily administration.

[0045] Compositions for use in accordance with the present invention (e.g., compositions for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or compositions for use in methods for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) comprise one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate. Methods of using the compositions are also provided herein.

[0046] In each of the aspects and embodiments of the invention described herein, rice bran may be replaced with any suitable grain (e.g., cereal grain) unless the context indicates otherwise. In such aspects and embodiments, the suitable grain may be the bran of any suitable grain. Suitable grains and suitable grain brans are discussed elsewhere herein.

[0047] Throughout the description and claims of this specification, the words "comprise" and "contain," and variations thereof, mean "including, but not limited to," and are not intended to (and do not) exclude other moieties, adjuncts, components, integers, or steps.

[0048] Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification should be understood to contemplate the plural as well as the singular, unless the context otherwise requires.

[0049] It should be understood that any feature, integer, characteristic, compound, chemical moiety, or chemical group described in connection with a particular aspect, embodiment, or example of the invention is applicable to any other aspect, embodiment, or example described herein, unless incompatible.

[0050] Various aspects of the invention are described in further detail below.

[0051] Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure Several Bacillus strains described in this application have been deposited with the Belgian Coordinated Collections of Micro-organisms (BCCM) Laboratoriumvoor Microbiologie - Bacterienverzameling (LMG), an international depository located at Universiteit Gent, KL Ledeganckstraat 35, 9000 Gent, Belgium. These deposits were made under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

[0052] Embodiments of the present invention are described in further detail herein with reference to the accompanying drawings. [Brief explanation of the drawings]

[0053] [Figure 1] Figure 1 shows the change in Freestyle Libre analysis between the first two weeks (weeks 0-2) and the last two weeks (weeks 5-6) of the observation period of the study. [Figure 2]Figure 2 shows the change from baseline in measured biomarkers of glycemic control and underlying diabetic exacerbation after 6 weeks of treatment (n=40). DETAILED DESCRIPTION OF THE INVENTION

[0054] The patent, scientific, and technical literature referred to herein constitutes the knowledge available to those skilled in the art at the time of filing. The entire disclosures of issued patents, published and pending patent applications, and other publications cited herein are incorporated by reference herein as if each were specifically and individually indicated to be incorporated by reference. In the case of conflicts, the present disclosure shall control.

[0055] Various aspects of the invention are described in further detail below.

[0056] The present invention is based on the surprising finding that (fermented) rice bran-based compositions (e.g., nutritional supplements) comprising Bacillus subtilis and Bacillus coagulans can be advantageously used to safely improve the cardiometabolic status of subjects, particularly those with type 2 diabetes and / or pre-diabetes.

[0057] The inventors have surprisingly shown that AB001, compared with placebo, has beneficial effects on glycemic control in patients with type 2 diabetes, as well as on biomarkers of inflammation (e.g., chronic systemic inflammation), insulin resistance, and beta-cell dysfunction. The inventors have therefore advantageously demonstrated that the composition described herein (AB001) can be used to stabilize glucose levels and / or improve glycemic control, particularly in subjects with type 2 diabetes and / or prediabetes. Advantageously, the compositions described herein can also be used to reduce insulin resistance, beta-cell dysfunction, and / or inflammation (e.g., chronic systemic inflammation and / or intestinal inflammation), particularly in subjects with type 2 diabetes and / or prediabetes. In some examples, the inflammation can be intestinal and / or pancreatic inflammation. In addition, the inventors have surprisingly observed that changes in lipid profiles and other metabolic syndrome parameters in patients with type 2 diabetes are more favorable with AB001 than with placebo. Advantageously, the compositions described herein may therefore be used for the treatment and / or prevention of symptoms of metabolic syndrome in subjects with type 2 diabetes and / or pre-diabetes.

[0058] Thus, provided herein is a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject.

[0059] Also provided is a method for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, comprising administering to the subject a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

[0060] Compositions used in accordance with the present invention Compositions used in the treatment and / or prevention of type 2 diabetes, prediabetes, and / or symptoms thereof in a subject and / or compositions used in methods of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject may be referred to as "MYRKL-S," "MYRKLS," "SB001," and / or "AB001."

[0061] AB001 comprises specially selected bacterial strains that preferably break down (e.g., metabolize) glucose and other carbohydrates and also reduce inflammation (in the intestine and / or pancreas). Without being bound by theory, AB001 is suggested to have four glucose-controlling effects, as discussed elsewhere herein.

[0062] In the body, complex carbohydrates reaching the small intestine must be hydrolyzed into monosaccharides such as glucose or galactose for transport across the intestinal mucosa. The term carbohydrate, particularly in biochemistry, is synonymous with saccharide. Carbohydrates (saccharides) can be divided into four chemical classes: monosaccharides, disaccharides, oligosaccharides, and polysaccharides. Generally, relatively small (low molecular weight) carbohydrates, monosaccharides and disaccharides, are broadly referred to as sugars. Those skilled in the art can easily identify carbohydrates using standard methods in the art. A non-limiting example of a carbohydrate is glucose. The classical pathway of glucose absorption is across the intestinal brush border membrane (BBM) and is thought to be primarily mediated by the sodium-dependent glucose transporter SGLT1 present at the apical end of enterocytes. This cotransporter (SGLT1) utilizes the electrochemical gradient of two sodium ions to transport one glucose molecule. Intracellular glucose is released into the interstitial space near capillaries by a mechanism thought to occur by facilitated diffusion via the glucose transporter GLUT2 present at the basolateral membrane of enterocytes.

[0063] Glucose is a six-carbon molecule with the chemical formula C6H12O6. It is a ubiquitous source of energy for all living organisms worldwide and is an essential fuel for both aerobic and anaerobic cellular respiration.

[0064] In some instances, the composition used in accordance with the present invention may be a glucose degrading composition.

[0065] As will be apparent to one of ordinary skill in the art, a "glucolytic (degrading) composition," as referred to herein, is a composition that breaks down glucose. In other words, a "glucolytic (degrading) composition," as used herein, is a composition that breaks down (e.g., biochemically) glucose. In the context of the present invention, the degradation (e.g., breakdown) of a substance (such as a carbohydrate, e.g., glucose) includes converting the substance into one or more other distinct substances. Furthermore, as will be apparent to one of ordinary skill in the art, "glucose degrading," as used herein, refers to the breaking down of glucose, and "glucose degradation," as used herein, refers to the breakdown of glucose. As will be apparent to one of ordinary skill in the art, the compositions described herein can be used in vitro or in vivo.

[0066] Glucose can be degraded (e.g., broken down) in a number of different ways. Of particular relevance in the context of the present invention is biochemical degradation. Thus, in one example, glucose can be biochemically degraded (e.g., broken down). As will be appreciated by those skilled in the art, the biochemical breakdown of a substance can involve enzyme-catalyzed reactions. Thus, in one example, glucose can be enzymatically degraded.

[0067] Advantageously, the compositions described herein may promote the degradation (e.g., breakdown) of glucose in the digestive tract and reduce glucose absorption through the intestinal tract. Similar effects and corresponding biochemical findings are seen when drugs belonging to the SGLT-1 inhibitor class (α-glucosidase inhibitors, e.g., acarbose or voglibose) are used in routine diabetes treatment. SGLT-1 inhibitors actively retain glucose molecules in the intestinal tract, where they are ultimately metabolized by resident bacteria in the large intestine, with methane gas being the end product. Advantageously, AB001 is suggested to have four glucose-controlling actions, as discussed above. Supplementation with AB001 is suggested to mean a reduction in glucose reaching the blood.

[0068] "Metabolism," as used herein, is a term used to describe all biochemical reactions involved in maintaining cells and organisms alive. For example, metabolism includes all biochemical reactions involved in converting one molecule into another (essentially to maintain a cell or organism alive). Metabolism includes processes for cell growth, reproduction, response to the environment, survival mechanisms, life support, and maintaining cellular structure and integrity. The biochemical reactions involved in metabolism utilize various enzymes.

[0069] In one example, glucose can be metabolized (i.e., glucose can be broken down by metabolism). Thus, in one example, a glucose metabolism composition is provided for use in accordance with the present invention (e.g., for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or for use in a method of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject), comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate (e.g., dextrin). As will be appreciated by those skilled in the art, metabolism can be enzymatic, and thus, in some examples, glucose can be enzymatically metabolized.

[0070] Metabolism can be divided into two categories: catabolism and anabolism. Catabolism involves a series of degradative biochemical reactions that break down complex molecules into smaller units, usually releasing energy in the process. For example, catabolism can be used to refer to any biochemical or enzymatic reaction involved in the breakdown of organic or inorganic substances such as proteins, sugars, fatty acids, etc. Anabolism involves a series of biochemical reactions that build or synthesize molecules from smaller units, usually requiring the input of energy (ATP) in the process. Thus, catabolism refers to the destructive biochemical reactions that occur within an organism, while metabolism refers to the entire suite of biochemical reactions within an organism, whether constructive or destructive.

[0071] In some instances, the breakdown of glucose by metabolism can be considered catabolism. Thus, in some instances, glucose can be catabolized (i.e., glucose can be broken down by catabolism). In further instances, glucose can be catabolized by enzymes.

[0072] Typically, in the context of the present invention, glucose is degraded in the subject's body. As a non-limiting example, glucose can be enzymatically destroyed, metabolized, or catabolized in the subject's body. When glucose degradation occurs in the subject's body (e.g., particularly in the duodenum and / or small intestine), this can be referred to as digestion. In some examples, glucose can be degraded by biochemical digestion.

[0073] Thus, in one example, provided herein is a glucose-degrading composition for use in accordance with the present invention, comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate (e.g., dextrin).

[0074] In another example, provided herein is a glucose metabolism composition for use in accordance with the present invention, comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate (e.g., dextrin).

[0075] As discussed elsewhere herein, compositions used in accordance with the present invention (e.g., compositions used to treat and / or prevent type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject, and / or compositions used in methods of treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject) comprise one or more Bacillus species. For example, compositions used in accordance with the present invention may comprise two or more Bacillus species. In another example, compositions used in accordance with the present invention may comprise three or more Bacillus species. In a further example, compositions used in accordance with the present invention may comprise four or more Bacillus species. In another example, compositions used in accordance with the present invention may comprise five or more Bacillus species. In a further example, compositions used in accordance with the present invention may comprise six or more Bacillus species. In another example, compositions used in accordance with the present invention may comprise seven or more Bacillus species.

[0076] As is known in the art, a genus is composed of several species. Thus, the genus "Bacillus" includes all species within the genus "Bacillus" known to those skilled in the art, including, but not limited to, Bacillusalkalophilus, Bacillusamyloliquefaciens, Bacillusbrevis, Bacilluscirculans, Bacillusclausii, Bacilluscoagulans, Bacillusfirmus, Bacilluslautus, Bacilluslentus, Bacilluslicheniformis, Bacillusmegaterium, Bacilluspumilus, Bacillusstearothermophilus, Bacillussubtilis, Bacillusvelezensis, BacillusspMT03, Bacillusatrophaeus, and Bacillusthuringiensis. It is recognized that the genus Bacillus is continually undergoing taxonomic reorganization. Thus, the genus is intended to include reclassified species, including, but not limited to, organisms such as B. stearothermophilus (now named "Geobacillus stearothermophilus"). The production of endospores that are resistant in the presence of oxygen is considered a defining feature of the genus Bacillus, although this characteristic also applies to the recently named species Alicyclobacillus, Amphibacillus, Aneurinibacillus, Anoxybacillus, Brevibacillus, Filobacillus, Gracilibacillus, Halobacillus, Paenibacillus, Salibacillus, Thermobacillus, Ureibacillus, and Virgibacillus.

[0077] Bacillus species are gram-positive bacteria classified as members of the family Bacillaceae, order Bacillales, and class Bacilli. As used herein, "Bacillus species" ("Bacillus sp.") refers to species of the genus "Bacillus."

[0078] Bacillus species found to be particularly important in the context of the present invention include Bacillus subtilis (B. subtilis) and Bacillus coagulans (B. coagulans). As described in the examples below, these bacteria are abundant in compositions used in accordance with the present invention and are particularly effective at breaking down (e.g., metabolizing) glucose, especially in the gastrointestinal tract.

[0079] Thus, in one example, the one or more Bacillus species is selected from B. subtilis and B. coagulans.

[0080] In another example, a composition for use in accordance with the present invention comprises B. subtilis and B. coagulans.

[0081] Any suitable B. subtilis and / or B. coagulans strain may be used in accordance with the present invention. Those skilled in the art will be able to readily identify suitable strains. Bacillus subtilis strain DFM 0326 (LMG P-32899), Bacillus subtilis strain DFM 1015 (LMG P-32900), and Bacillus coagulans strain DFM 0705 (LMG P-32921) have been found to be particularly important in the context of the present invention.

[0082] The Bacillus subtilis strain deposited under LMG-P accession number 32899 may be referred to herein as "DFM 0326" or "strain DFM 0326." Bacillus subtilis strain DFM 0326 (deposited under LMG-P accession number 32899) was deposited under the Budapest Treaty of 1977 on November 22, 2022, with the Belgian Coordinated Collections of Micro-organisms (BCCM), Laboratoriumvoor Microbiologie - Bacterienverzameling (LMG), Universiteit Gent, KLLedeganckstraat 35, 9000 Gent, Belgium.

[0083] The Bacillus subtilis strain deposited under LMG-P accession number 32900 may be referred to herein as "DFM 1015" or "strain DFM 1015." Bacillus subtilis strain DFM 1015 (deposited under LMG-P accession number 32900) was deposited under the Budapest Treaty of 1977 on November 22, 2022, with the Belgian Coordinated Collections of Micro-organisms (BCCM), Laboratoriumvoor Microbiologie - Bacterienverzameling (LMG), Universiteit Gent, KLLedeganckstraat 35, 9000 Gent, Belgium.

[0084] The Bacillus coagulans strain deposited under LMG-P accession number 32921 may be referred to herein as "DFM 0705" or "strain DFM 0705." Bacillus coagulans strain DFM 0705 (deposited under LMG-P accession number 32921) was deposited under the Budapest Treaty of 1977 on December 14, 2022 with the Belgian Coordinated Collections of Micro-organisms (BCCM), Laboratoriumvoor Microbiologie - Bacterienverzameling (LMG), Universiteit Gent, KLLedeganckstraat 35, 9000 Gent, Belgium.

[0085] Thus, in some examples, the B. subtilis species may be selected from the group consisting of Bacillus subtilis strain DFM 0326 (LMG P-32899) and Bacillus subtilis strain DFM 1015 (LMG P-32900), and / or the B. coagulans species may be Bacillus coagulans strain DFM 0705 (LMG P-32921).

[0086] Thus, in an example where the composition comprises B. subtilis and B. coagulans, the B. subtilis species may be Bacillus subtilis strain DFM 0326 (LMG P-32899) and the B. coagulans species may be Bacillus coagulans strain DFM 0705 (LMG P-32921).

[0087] In another example where the composition comprises B. subtilis and B. coagulans, the B. subtilis species can be Bacillus subtilis strain DFM 1015 (LMG P-32900) and the B. coagulans species can be Bacillus coagulans strain DFM 0705 (LMG P-32921).

[0088] In some examples, compositions according to the invention may include B. subtilis strain DFM 0326 (LMG P-32899) in combination with Bacillus subtilis strain DFM 1015 (LMG P-32900).

[0089] In another example where a composition includes B. subtilis and B. coagulans, the composition may include B. subtilis strain DFM 0326 (LMG P-32899), and Bacillus subtilis strain DFM 1015 (LMG P-32900), and B. coagulans species strain DFM 0705 (LMG P-32921). Note that the composition used in the examples provided herein includes B. subtilis strain DFM 0326 (LMG P-32899), and Bacillus subtilis strain DFM 1015 (LMG P-32900), and B. coagulans strain DFM 0705 (LMG P-32921).

[0090] Thus, compositions for use in accordance with the present invention may include Bacillus subtilis strain DFM 0326 (LMG P-32899), Bacillus subtilis strain DFM 1015 (LMG P-32900), and Bacillus coagulans strain DFM 0705 (LMG P-32921).

[0091] The inventors have also identified several other bacterial species that are particularly relevant in the context of the present invention. Thus, in one example, a composition for use in accordance with the present invention may further comprise one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillusvelezensis, Bacillus sp MT 03, Bacillus lusatrophaeus, and Pediococcus pentosaceus. In one example, a composition for use in accordance with the present invention may further comprise one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillusvelezensis, Bacillus sp MT 03, Bacillus lusatrophaeus, Pediococcus pentosaceus, and the genus Saccharomycetes (e.g., Brettanomycesbruxellensis and / or Millerozoma farinosa). In another example, a composition for use in accordance with the present invention may further comprise two or more, or three or more, or four or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillusvelezensis, Bacillus sp MT 03, Bacillus lusatrophaeus, and Pediococcus pentosaceus. In another example, a composition for use in accordance with the present invention may further comprise two or more, or three or more, or four or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillusvelezensis, Bacillus sp MT 03, Bacillus lusatrophaeus, Pediococcus pentosaceus, and a member of the genus Saccharomycetes (e.g., Brettanomycesbruxellensis and / or Millerozoma farinosa).

[0092] In another example, a composition for use in accordance with the present invention may further comprise Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus. In another example, a composition for use in accordance with the present invention may further comprise Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and a genus of Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozoma farinosa).

[0093] The bacterial strains included in the compositions described herein have been identified to favorably and efficiently metabolize glucose. Advantageously, it has been suggested that one result of AB001 supplementation is a reduction in glucose absorption through the intestinal tract. Therefore, it has been suggested that AB001 supplementation means a reduction in glucose reaching the blood.

[0094] Compositions used in accordance with the present invention include a suitable amount or concentration of one or more Bacillus species. The amount or concentration of one or more Bacillus species may therefore be described by reference to the number of colony forming units (cfu) per gram of composition (cfu / g) or by the total number of colony forming units (cfu) per dose of the composition (in other words, cfu per effective dose). As will be apparent to those skilled in the art, and as described in more detail elsewhere herein, a dose may include one or more dosage units (e.g., two dosage units). In instances where multiple dosage units are used to provide an effective dose, cfu / dose corresponds to the total cfu of all the multiple dosage units.

[0095] In one example, the compositions described herein contain between about 10,000 cfu / g and about 1 x 10 Bacillus bacteria.8 Contains cfu / g of Bacillus bacteria.

[0096] In one example, the compositions described herein contain at least about 1×10 8 In another example, the compositions described herein contain at least about 1 x 10 cfu / g of Bacillus bacteria. 7 In another example, the compositions described herein contain at least about 1 x 10 cfu / g of Bacillus bacteria. 6 In a further example, the compositions described herein contain at least about 1 x 10 cfu / g of Bacillus bacteria. 5 Contains cfu / g of Bacillus bacteria.

[0097] In one example, the compositions described herein contain at least about 10,000 cfu / g (i.e., at least about 1.0 x 10 4 cfu / g) of Bacillus genus bacteria.

[0098] In another example, the compositions described herein may have an immunization rate of at least about 11,000 cfu / g (i.e., at least about 1.1 x 10 4 cfu / g). In a further example, the compositions described herein contain at least about 12,000 cfu / g (i.e., at least about 1.2 x 10 4 cfu / g) of Bacillus genus bacteria.

[0099] In another example, the compositions described herein have an immunization rate of at least about 13,000 cfu / g (i.e., at least about 1.3×10 4 cfu / g) of Bacillus genus bacteria.

[0100] In another example, the compositions described herein have an immunization rate of at least about 14,000 cfu / g (i.e., at least about 1.4×10 4 cfu / g). In another example, the compositions described herein contain at least about 15,000 cfu / g (i.e., at least about 1.5 x 104 cfu / g) of Bacillus genus bacteria.

[0101] In one example, the compositions described herein contain about 1×10 8 In another example, the compositions described herein contain about 1 x 10 cfu / g of Bacillus bacteria. 7 In another example, the compositions described herein contain about 1 x 10 cfu / g of Bacillus bacteria. 6 cfu / g of Bacillus bacteria. In a further example, the compositions described herein contain about 1 x 10 5 Contains cfu / g of Bacillus bacteria.

[0102] In one example, the compositions described herein contain about 10,000 cfu / g (i.e., about 1.0 x 10 4 cfu / g) of Bacillus genus bacteria.

[0103] In another example, the compositions described herein contain about 11,000 cfu / g (i.e., about 1.1 x 10 4 cfu / g). In a further example, the compositions described herein contain about 12,000 cfu / g (i.e., about 1.2 x 10 4 cfu / g) of Bacillus genus bacteria.

[0104] In another example, the compositions described herein contain about 13,000 cfu / g (i.e., about 1.3 x 10 4 cfu / g) of Bacillus genus bacteria.

[0105] In another example, the compositions described herein contain about 14,000 cfu / g (i.e., about 1.4×10 4 cfu / g). In a further example, the compositions described herein contain about 15,000 cfu / g (i.e., about 1.5 x 10 4 cfu / g) of Bacillus genus bacteria.

[0106] Those skilled in the art can readily determine the amount or concentration of bacteria present in a composition using routine methods known in the art. For example, the total viable count (TVC) of live Bacillus cells can be determined by established culture methods based on specific Bacillus media, such as ChromeSelect agar. Alternative methods are known in the art.

[0107] In one example, the compositions described herein contain between about 5,000 cfu of Bacillus bacteria / dose and about 1 x 10 8 Contains cfu of Bacillus bacteria / dose.

[0108] In one example, the compositions described herein contain at least about 1×10 8 In another example, the compositions described herein comprise at least about 1 x 10 cfu of Bacillus bacteria per dose. 7 In another example, the compositions described herein comprise at least about 1 x 10 cfu of Bacillus bacteria per dose. 6 Contains cfu of Bacillus bacteria / dose.

[0109] In a further example, the compositions described herein contain at least about 1×10 5 Contains cfu of Bacillus bacteria / dose.

[0110] In one example, the compositions described herein contain at least about 5,000 cfu (i.e., at least about 0.5×10 4 cfu) of Bacillus spp. / dose.

[0111] In another example, the compositions described herein contain at least about 10,000 cfu (i.e., at least about 1.0 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain at least about 11,000 cfu (i.e., at least about 1.1 x 10 4cfu) of Bacillus bacteria / dose. In a further example, the compositions described herein contain at least about 12,000 cfu (i.e., at least about 1.2 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain at least about 13,000 cfu (i.e., at least about 1.3 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain at least about 14,000 cfu (i.e., at least about 1.4 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain at least about 15,000 cfu (i.e., at least about 1.5×10 4 cfu) of Bacillus spp. / dose.

[0112] In one example, the compositions described herein contain about 1×10 8 In another example, the compositions described herein contain about 1 x 10 cfu of Bacillus bacteria per dose. 7 In another example, the compositions described herein contain about 1 x 10 cfu of Bacillus bacteria per dose. 6 Contains cfu of Bacillus bacteria / dose.

[0113] In a further example, the compositions described herein may contain about 1×10 5 Contains cfu of Bacillus bacteria / dose.

[0114] In one example, the compositions described herein contain about 5,000 cfu (i.e., about 0.5×10 4 cfu) of Bacillus bacteria / dose. In one example, the compositions described herein contain about 10,000 cfu (i.e., about 1.0 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain about 11,000 cfu (i.e., about 1.1 x 10 4cfu) of Bacillus bacteria / dose. In a further example, the compositions described herein contain about 12,000 cfu (i.e., about 1.2 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain about 13,000 cfu (i.e., about 1.3 x 10 4 cfu) of Bacillus bacteria / dose. In another example, the compositions described herein contain about 14,000 cfu (i.e., about 1.4×10 4 cfu) of Bacillus bacteria / dose. In a further example, the compositions described herein contain about 15,000 cfu (i.e., about 1.5×10 4 cfu) of Bacillus spp. / dose.

[0115] As will be apparent to those skilled in the art, the amount or concentration of one or more Bacillus species in a composition can be comprised of any one species of Bacillus or any combination of Bacillus species. For example, the amount or concentration of one or more Bacillus species in a composition can be comprised entirely of one Bacillus species (e.g., the concentration can be comprised entirely of B. subtilis or entirely of B. coagulans). Alternatively, the amount or concentration of one or more Bacillus species in a composition can be comprised of two or more, three or more, four or more, or five or more Bacillus species (e.g., the concentration can be comprised of B. subtilis and B. coagulans). Thus, in one example, the amount or concentration of one or more Bacillus species in a composition refers to the amount or concentration of the combination of Bacillus species present.

[0116] In some examples, the compositions provided herein include a suitable concentration of bacteria, where the concentration ratio is comprised of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans). In one example, the compositions described herein contain about 10,000 cfu / g (i.e., about 1.0 x 10 4 cfu / g) bacteria ~ approx. 1 x 108 cfu / g of bacteria. In one example, the compositions described herein contain at least about 1 x 10 8 In another example, the compositions described herein contain at least about 1 x 10 cfu / g of bacteria. 7 In another example, the compositions described herein contain at least about 1 x 10 cfu / g of bacteria. 6 In a further example, the compositions described herein contain at least about 1 x 10 cfu / g of bacteria. 5 cfu / g of bacteria. In one example, the compositions described herein contain at least about 10,000 cfu / g (i.e., at least about 1.0 x 10 4 In another example, the compositions described herein contain at least about 11,000 cfu / g (i.e., at least about 1.1 x 10 4 In a further example, the compositions described herein contain at least about 12,000 cfu / g (i.e., at least about 1.2 x 10 4 In another example, the compositions described herein contain at least about 13,000 cfu / g (i.e., at least about 1.3 x 10 4 In another example, the compositions described herein contain at least about 14,000 cfu / g (i.e., at least about 1.4 x 10 4 In another example, the compositions described herein contain at least about 15,000 cfu / g (i.e., at least about 1.5 x 10 4 cfu / g). In one example, the compositions described herein contain about 1 x 10 8 In another example, the compositions described herein contain about 1 x 10 cfu / g of bacteria. 7 In another example, the compositions described herein contain about 1 x 10 cfu / g of bacteria. 6 cfu / g of bacteria. In a further example, the compositions described herein contain about 1 x 10 5 cfu / g of bacteria. In one example, the compositions described herein contain about 10,000 cfu / g (i.e., about 1.0 x 10 4In another example, the compositions described herein contain about 11,000 cfu / g (i.e., about 1.1 x 10 4 cfu / g). In a further example, the compositions described herein contain about 12,000 cfu / g (i.e., about 1.2 x 10 4 In another example, the compositions described herein contain about 13,000 cfu / g (i.e., about 1.3 x 10 4 In another example, the compositions described herein contain about 14,000 cfu / g (i.e., about 1.4 x 10 4 cfu / g). In a further example, the compositions described herein contain about 15,000 cfu / g (i.e., about 1.5 x 10 4 cfu / g) of bacteria.

[0117] In some examples, the compositions described herein include a suitable amount of bacteria, the proportion of which comprises one or more Bacillus species (e.g., B. subtilis and / or B. coagulans). In one example, the compositions described herein contain between about 5,000 cfu of bacteria / dose and about 1 x 10 8 In one example, the compositions described herein contain at least about 1 x 10 cfu of bacteria / dose. 8 In another example, the compositions described herein comprise at least about 1 x 10 cfu of bacteria / dose. 7 In another example, the compositions described herein comprise at least about 1 x 10 cfu of bacteria / dose. 6 In a further example, the compositions described herein comprise at least about 1 x 10 cfu of bacteria / dose. 5 cfu of bacteria / dose. In one example, the compositions described herein contain at least about 5,000 cfu (i.e., at least about 0.5 x 10 4 In another example, the compositions described herein contain at least about 10,000 cfu (i.e., at least about 1.0 x 10 4In another example, the compositions described herein contain at least about 11,000 cfu (i.e., at least about 1.1 x 10 4 cfu) of bacteria / dose. In a further example, the compositions described herein contain at least about 12,000 cfu (i.e., at least about 1.2 x 10 4 In another example, the compositions described herein contain at least about 13,000 cfu (i.e., at least about 1.3 x 10 4 In another example, the compositions described herein contain at least about 14,000 cfu (i.e., at least about 1.4 x 10 4 In another example, the compositions described herein contain at least about 15,000 cfu (i.e., at least about 1.5 x 10 4 cfu) of bacteria / dose. In one example, the compositions described herein contain about 1 x 10 8 In another example, the compositions described herein contain about 1 x 10 cfu of bacteria / dose. 7 In another example, the compositions described herein contain about 1 x 10 cfu of bacteria / dose. 6 In a further example, the compositions described herein comprise about 1 x 10 cfu of bacteria / dose. 5 cfu of bacteria / dose. In one example, the compositions described herein contain about 5,000 cfu (i.e., about 0.5×10 4 cfu) of bacteria / dose. In one example, the compositions described herein contain about 10,000 cfu (i.e., about 1.0 x 10 4 In another example, the compositions described herein contain about 11,000 cfu (i.e., about 1.1 x 10 4 cfu) of bacteria / dose. In a further example, the compositions described herein contain about 12,000 cfu (i.e., about 1.2 x 10 4 In another example, the compositions described herein contain about 13,000 cfu (i.e., about 1.3 x 10 4cfu) of bacteria / dose. In another example, the compositions described herein contain about 14,000 cfu (i.e., about 1.4 x 10 4 cfu) of bacteria / dose. In a further example, the compositions described herein contain about 15,000 cfu (i.e., about 1.5 x 10 4 cfu) of bacteria / dose.

[0118] As will be apparent to one of skill in the art, the amount or concentration of bacteria (proportion comprised of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans)) may further include any suitable single species or any combination of species. For example, the amount or concentration of bacteria (proportion comprised of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans)) may further include a proportion of one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus. For example, the amount or concentration of bacteria (proportion comprised of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans)) may further include a proportion of one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus. 03, Bacillus atrophaeus, Pediococcus pentosaceus, and a proportion of one or more bacterial species selected from the group consisting of the genus Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozomafarinosa).

[0119] In one example, one or more Bacillus species are genetically modified. As used herein, "genetic modification" and "genetic engineering" refer to the direct manipulation (e.g., modification) of one or more genes, for example, using recombinant DNA technology. While humans have traditionally manipulated genomes indirectly by controlling reproduction and selecting offspring with desired traits, genetic engineering involves the direct manipulation (e.g., modification) of one or more genes. For example, a gene from another species may be added to the genome of an organism to confer a desired phenotype on that organism.

[0120] In other examples, the one or more Bacillus species are not genetically modified, i.e., the one or more Bacillus species may occur in nature.

[0121] One or more Bacillus species may be present in a microbial consortium. A "microbial consortium," as used herein, refers to a group of microorganisms (e.g., bacteria), where the group includes two or more distinct microorganisms (e.g., two or more bacteria that may be from the same species (e.g., two or more distinct strains) or distinct species (e.g., two or more distinct species).

[0122] The microbial consortium can be a naturally occurring microbial consortium (e.g., a consortium naturally produced during fermentation of a grain, such as rice bran). In other words, one or more Bacillus species can be part of the composition due to the presence of a grain, such as fermented rice bran (with a naturally produced microbial consortium) in the composition. Thus, one or more Bacillus species can be natural components of a grain, such as fermented rice bran.

[0123] The inventors have found that Bacillus subtilis strain DFM 0326 (LMG P-32899), Bacillus subtilis strain DFM 1015 (LMG P-32900), and Bacillus coagulans strain DFM 0705 (LMG P-32921) can be isolated from the compositions (specifically fermented rice bran) used in the examples provided herein. Thus, B. subtilis strain DFM 0326 (LMG P-32899) and Bacillus subtilis strain DFM 1015 (LMG P-32900), as well as B. coagulans strain DFM 0705 (LMG P-32921), can be part of the compositions described herein due to the presence of fermented grains (with naturally occurring microbial consortia) such as rice bran in the compositions.

[0124] In one example, one or more Bacillus species selected from B. subtilis and B. coagulans may be (part of) a naturally occurring microbial consortium (e.g., a consortium naturally produced during the fermentation of a grain, such as rice bran). In other words, one or more Bacillus species selected from B. subtilis and B. coagulans may be part of a composition due to the presence of a grain, such as fermented rice bran (with a naturally produced microbial consortium) in the composition. Thus, one or more Bacillus species selected from B. subtilis and B. coagulans may be a natural component of a grain, such as fermented rice bran.

[0125] In another example, one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus may be (part of) a naturally occurring microbial consortium (e.g., a consortium formed naturally during the fermentation of a grain such as rice bran). In other words, one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus may be part of the composition due to the presence of a grain such as fermented rice bran (with a naturally occurring microbial consortium) in the composition. Thus, one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillusvelezensis, Bacillus sp MT 03, acillusatrophaeus, and Pediococcus pentosaceus may be natural constituents of grains such as fermented rice bran.

[0126] In another example, one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genus Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozoma farinose) may be (part of) a naturally occurring microbial consortium (e.g., a consortium formed naturally during the fermentation of grains such as rice bran). In other words, one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genus Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozomafarinosa) may be part of the composition due to the presence of a grain such as fermented rice bran (with a naturally occurring microbial consortium) in the composition. Thus, Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genus Saccharomycetes (e.g., Brettanomyces One or more bacterial species selected from the group consisting of: Saccharomyces cerevisiae, ...

[0127] In another example, one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) and one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus may be (part of) a naturally occurring microbial consortium (e.g., a consortium formed naturally during the fermentation of grains such as rice bran). In other words, one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) and one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus may be part of the composition due to the presence of a grain such as fermented rice bran (with a naturally occurring microbial consortium) in the composition. Thus, one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) and one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus may be natural components of a grain such as fermented rice bran.

[0128] In another example, one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) and one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus satrophaeus, Pediococcus pentosaceus, and the genera Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozoma farinosa) can be (part of) a naturally occurring microbial consortium (e.g., a consortium formed naturally during the fermentation of grains such as rice bran). In other words, one or more species of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans), and one or more species of bacteria selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genera Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozoma farinosa) may be part of the composition due to the presence of a grain (with a naturally occurring microbial consortium) such as fermented rice bran in the composition. Thus, one or more species of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans), as well as one or more species of bacteria selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, Pediococcus pentosaceus, and the genera Saccharomycetes (e.g., Brettanomyces bruxellensis and / or Millerozoma farinose), may be natural constituents of grains such as fermented rice bran.

[0129] As will be apparent to those skilled in the art, the compositions used in accordance with the present invention are not limited to naturally occurring microbial consortia. Thus, the microbial consortia discussed above may be artificially created, for example, by combining one or more bacterial isolates with each other. In one example, therefore, one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) may be added individually to the composition.

[0130] Compositions used in accordance with the present invention (e.g., compositions used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject and / or compositions used in methods of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) may therefore also include grains such as rice bran (e.g., fermented rice bran), as discussed in more detail elsewhere herein. Fermented rice bran may, for example, be a natural source of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans) present in the composition.

[0131] As described elsewhere herein, compositions for use in accordance with the present invention (e.g., compositions for use in treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject, and / or compositions for use in methods for treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject) comprise L-cysteine in combination with one or more species of Bacillus bacteria (e.g., the bacteria are added to the composition as a bacterial supplement or as a natural product, for example, as part of fermented rice bran).

[0132] L-cysteine (L-cys) is a non-essential amino acid and therefore one of the building blocks required for protein synthesis. L-cysteine contains sulfur in the form of a thiol group (-SH) at the end of its side chain. This -SH group is responsible for the amino acid's high reactive capacity and therefore many of its biological functions in the human body. L-cysteine occupies a key position in the sulfur metabolism of all living organisms and is used in the synthesis of proteins, glutathione, biotin, lipoic acid, methionine, and other sulfur-containing metabolites. In addition, L-cysteine serves as a precursor for the biosynthesis of coenzyme A. L-cysteine biosynthesis has been extensively studied in bacteria, particularly enterobacteria. The amino acid L-cysteine is not only biologically important but also economically important. For example, L-cysteine is used as a food additive (especially in the bakery and confectionery industries), as a starting material for cosmetics, and as a starting material for the preparation of active ingredients (especially N-acetyl-cysteine and S-carboxymethyl-cysteine).

[0133] As will be apparent to those skilled in the art, references herein to L-cysteine refer to the amino acid L-cysteine in any suitable form, and thus the term "L-cysteine" encompasses the free form of L-cysteine as well as L-cysteine salts.

[0134] In the context of the present invention, in one example, L-cysteine can be in the free form, its salt, or a mixture thereof. Examples of salts include sulfate, hydrochloride, carbonate, ammonium salt, sodium salt, and potassium salt. In one example, L-cysteine is in the form of crystalline pieces larger than 0.1 mm.

[0135] L-cysteine is available from several sources and can be easily obtained by those skilled in the art. Furthermore, those skilled in the art can easily detect the presence of L-cysteine in a substance (e.g., the compositions described herein) using methods known in the art. For example, L-cysteine crystals can be macroscopically detected as white particles in the compositions described herein. HPLC, i.e., high performance liquid chromatography, using an Inertsil ODs-3 column is an established method for detecting L-cysteine.

[0136] L-cysteine can be obtained industrially by hydrolysis of animal materials such as poultry feathers or pig hair. In contrast, synthetic L-cysteine can be obtained by fermentation of genetically modified E. coli or Pseudomonas thiazolinophilum. Thus, in some instances, L-cysteine is of animal origin. In other instances, L-cysteine is of synthetic origin.

[0137] In one example, the L-cysteine is derived from a plant.

[0138] In some examples, the compositions described herein may include an L-cysteine derivative instead of, or in addition to, L-cysteine.

[0139] L-cysteine derivatives are well known to those skilled in the art.N-acetylcysteine (NAC) is the N-acetylated form of amino acid L-cysteine, and is therefore an example of L-cysteine derivatives.Therefore, in some examples, the compositions described herein can contain N-acetylcysteine (NAC).NAC is easily available in the art.

[0140] L-cysteine is present in the compositions described herein at any suitable concentration or amount. The amount or concentration of L-cysteine can therefore be described by reference to the % w / w of the composition or by the total weight (e.g., mg) per dose of the composition (in other words, the weight per effective dose). As will be apparent to those skilled in the art and as described in more detail elsewhere herein, a dose can include one or more dosage units (e.g., two dosage units). In instances where multiple dosage units are used to provide an effective dose, the weight per dose corresponds to the total weight of L-cysteine across all of the multiple dosage units.

[0141] In one example, the composition comprises about 10% w / w L-cysteine to about 40% w / w L-cysteine.

[0142] In another example, the composition comprises about 15% to about 35% w / w L-cysteine, hi another example, the composition comprises about 20% to about 30% w / w L-cysteine.

[0143] In one example, the composition comprises at least about 10% w / w L-cysteine.

[0144] In another example, the composition comprises at least about 15% w / w L-cysteine.

[0145] In a further example, the composition comprises at least about 20% w / w L-cysteine.

[0146] In another example, the composition comprises at least about 25% w / w L-cysteine. In another example, the composition comprises at least about 30% w / w L-cysteine. In a further example, the composition comprises at least about 35% w / w L-cysteine. In a further example, the composition comprises at least about 40% w / w L-cysteine.

[0147] In some examples, the composition comprises about 10% w / w L-cysteine.

[0148] In another example, the composition comprises about 15% w / w L-cysteine.

[0149] In another example, the composition comprises about 20% w / w L-cysteine.

[0150] In a further example, the composition comprises about 25% w / w L-cysteine. In another example, the composition comprises about 30% w / w L-cysteine. In another example, the composition comprises about 35% w / w L-cysteine. In a further example, the composition comprises about 40% w / w L-cysteine.

[0151] In one example, the composition contains about 38 mg of L-cysteine per dose to about 200 mg of L-cysteine per dose.

[0152] In one example, the composition comprises at least about 38 mg of L-cysteine per dose.

[0153] In one example, the composition contains at least about 50 mg of L-cysteine per dose. In one example, the composition contains at least about 75 mg of L-cysteine per dose. In one example, the composition contains at least about 100 mg of L-cysteine per dose.

[0154] In one example, the composition comprises at least about 150 mg of L-cysteine per dose.

[0155] In one example, the composition comprises at least about 160 mg of L-cysteine per dose. In one example, the composition comprises at least about 180 mg of L-cysteine per dose.

[0156] In one example, the composition contains about 38 mg of L-cysteine per dose.

[0157] In one example, the composition contains about 50 mg of L-cysteine per dose. In one example, the composition contains about 75 mg of L-cysteine per dose. In one example, the composition contains about 100 mg of L-cysteine per dose.

[0158] In one example, the composition contains about 150 mg of L-cysteine per dose.

[0159] In one example, the composition contains about 160 mg of L-cysteine per dose. In one example, the composition contains about 180 mg of L-cysteine per dose.

[0160] As described elsewhere herein, compositions for use in accordance with the present invention (e.g., compositions for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or compositions for use in methods for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) comprise L-cysteine in combination with one or more Bacillus species and a high molecular weight, low osmolality carbohydrate (e.g., dextrin).

[0161] As used herein, "high molecular weight low osmolality carbohydrate" refers to a carbohydrate having a molecular weight of about 500,000 g / mol to about 700,000 g / mol, wherein the carbohydrate has a low osmolality. A person skilled in the art would be readily able to identify suitable carbohydrates with low osmolality using routine tests known in the art. For the avoidance of doubt, as used herein, a low osmolality carbohydrate is a carbohydrate that has a glycogen recovery that is about 50% higher than maltodextrin.

[0162] The high molecular weight low osmolality carbohydrate is present in the compositions described herein at a suitable concentration or amount. The amount or concentration of the high molecular weight low osmolality carbohydrate can therefore be described by reference to the % w / w of the composition or by the total weight (e.g., mg) per dose of the composition (in other words, the weight per effective dose). As will be apparent to those skilled in the art and as described in more detail elsewhere herein, a dose can include one or more dosage units (e.g., two dosage units). In instances where multiple dosage units are used to provide an effective dose, the weight per dose corresponds to the total weight of the high molecular weight low osmolality carbohydrate of all multiple dosage units.

[0163] In one example, the composition comprises about 0.5% w / w to about 5% w / w of a high molecular weight, low osmolality carbohydrate.

[0164] In another example, the composition comprises about 0.5% w / w to about 3% w / w of a high molecular weight, low osmolality carbohydrate. In one example, the composition comprises about 0.5% w / w to about 2% w / w of a high molecular weight, low osmolality carbohydrate.

[0165] In one example, the composition comprises at least about 0.5% w / w of a high molecular weight, low osmolality carbohydrate.

[0166] In another example, the composition comprises at least about 1% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises at least about 1.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises at least about 2% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises at least about 2.5% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises at least about 3% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises at least about 3.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises at least about 4% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises at least about 4.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises at least about 5% w / w of high molecular weight, low osmolarity carbohydrates.

[0167] In one example, the composition comprises about 0.5% w / w of a high molecular weight, low osmolality carbohydrate.

[0168] In another example, the composition comprises about 1% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises about 1.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises about 2% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises about 2.5% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises about 3% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises about 3.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises about 4% w / w of high molecular weight, low osmolarity carbohydrates. In another example, the composition comprises about 4.5% w / w of high molecular weight, low osmolarity carbohydrates. In a further example, the composition comprises about 5% w / w of high molecular weight, low osmolarity carbohydrates.

[0169] In one example, the composition comprises about 2 mg to about 50 mg of high molecular weight, low osmolality carbohydrate per dose.

[0170] In one example, the composition comprises at least about 2 mg of high molecular weight, low osmolality carbohydrate per dose.

[0171] In one example, the composition comprises at least about 4 mg of high molecular weight, low osmolality carbohydrate per dose.

[0172] In one example, the composition comprises at least about 10 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 15 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 20 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 25 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 30 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 35 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 40 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition comprises at least about 45 mg of high molecular weight, low osmolarity carbohydrate per dose.

[0173] In one example, the composition comprises about 2 mg of high molecular weight, low osmolality carbohydrate per dose.

[0174] In one example, the composition comprises about 4 mg of high molecular weight, low osmolality carbohydrate per dose.

[0175] In one example, the composition contains about 10 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 15 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 20 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 25 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 30 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 35 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 40 mg of high molecular weight, low osmolarity carbohydrate per dose. In one example, the composition contains about 45 mg of high molecular weight, low osmolarity carbohydrate per dose.

[0176] In one example, the high molecular weight low osmolality carbohydrate is dextrin.

[0177] Dextrin is a generic term applied to various products obtained by heating starch in the presence of small amounts of water and acid. Dextrins are a group of low-molecular-weight carbohydrates resulting from the hydrolysis of starch or glycogen. "Dextrin" refers to a polymer of glucose resulting from the hydrolysis of starch (or glycogen) and contains glucose units linked together primarily by α-1,4 linkages. In addition to α-1,4 linkages, a proportion of α-1,6 linkages may be present in a particular dextrin, the amount depending on the starch starting material. Because the biodegradation rate of α-1,6 linkages is typically slower than that of α-1,4 linkages, for many applications, the proportion of α-1,6 linkages is preferably less than 10%, more preferably less than 5%. In some instances, dextrins are therefore mixtures of polymers of D-glucose units linked by α-(1→4) glycosidic or α-(1→6) glycosidic linkages.

[0178] Dextrins can be produced from starch using enzymes such as amylases, as occurs during human digestion, malting, and mashing, or by hot air drying (pyrolysis or roasting) under acidic conditions. The latter method is used in industry. Thermally produced dextrins are also known as pyrodextrins. Typically, dextrins are produced by hydrolysis of starch obtained from various natural sources, such as wheat, rice, corn, maize, and tapioca.

[0179] Dextrins are typically white, yellow, or brown powders that are partially or completely water-soluble and provide low-viscosity, optically active solutions. Dextrins are available from several sources and can be readily sourced by those skilled in the art. Those skilled in the art can readily identify suitable dextrins for use in the context of the present invention. Those skilled in the art can readily detect the presence of dextrin in a substance (e.g., a composition described herein) using methods known in the art. For example, most dextrins can be detected using iodine solution.

[0180] The term "dextrin" includes pyrodextrin, digestible dextrin, and hydrogenated products thereof, as well as derivatives thereof. The term "dextrin derivative" as used herein refers to a substance obtained by chemically or enzymatically treating dextrin, and includes, for example, the above-mentioned polydextrose, branched dextrin obtained by the action of glycosyltransferase on dextrin, and cyclodextrin obtained by the action of cyclodextrin-producing enzyme on starch.

[0181] In some instances, the dextrin is enzymatically treated.

[0182] In one example, the dextrin, and therefore the high molecular weight, low osmolality carbohydrate, is cluster dextrin (also known as highly branched cyclic dextrin). Cluster dextrin is a maltodextrin with a high molecular weight but a narrow molecular weight range, soluble, and near-zero osmolality. Typical high molecular weight carbohydrates are less soluble than cluster dextrin. Meanwhile, low molecular weight carbohydrates, like pure glucose solutions, exhibit higher osmolality. This slows their descent into the small intestine, similar to other digestible ingredients typically found in sports drinks. Cluster dextrin reaches the small intestine more quickly, resulting in faster sustained release. Solubility is central to cluster dextrin's function. High molecular weight cluster dextrin also degrades slowly, thereby balancing insulin secretion and lipid breakdown. Cluster dextrin is manufactured by Glico Nutrition.

[0183] In some instances, the dextrin, and thus the high molecular weight, low osmolality carbohydrate, is derived from wheat or corn (eg, in some instances, the dextrin is wheat dextrin or corn dextrin).

[0184] In some instances, the dextrin is enzyme-treated wheat or corn.

[0185] In one example, the dextrin is derived from wheat (e.g., wheat dextrin). For example, the dextrin can be wheat dextrin powder (such as Surbex Nutri-Fiber wheat dextrin powder, which is a soluble, non-viscous fiber).

[0186] In another example, the dextrin is derived from corn (eg, corn dextrin).

[0187] In a particular example, the dextrin is hydrolyzed corn dextrin (eg, Vitargo).

[0188] In some examples, dextrin is present in the compositions described herein at any suitable concentration or amount.

[0189] In one example, the composition comprises about 0.5% w / w to about 5% w / w dextrin.

[0190] In another example, the composition comprises from about 0.5% w / w to about 3% w / w of dextrin, hi another example, the composition comprises from about 0.5% w / w to about 2% w / w of dextrin.

[0191] In one example, the composition comprises at least about 0.5% w / w dextrin.

[0192] In another example, the composition comprises at least about 1% w / w dextrin. In another example, the composition comprises at least about 1.5% w / w dextrin. In a further example, the composition comprises at least about 2% w / w dextrin. In another example, the composition comprises at least about 2.5% w / w dextrin. In another example, the composition comprises at least about 3% w / w dextrin. In another example, the composition comprises at least about 3.5% w / w dextrin. In a further example, the composition comprises at least about 4% w / w dextrin. In another example, the composition comprises at least about 4.5% w / w dextrin. In a further example, the composition comprises at least about 5% w / w dextrin.

[0193] In one example, the composition comprises about 0.5% w / w dextrin.

[0194] In another example, the composition comprises about 1% w / w dextrin. In another example, the composition comprises about 1.5% w / w dextrin. In a further example, the composition comprises about 2% w / w dextrin. In another example, the composition comprises about 2.5% w / w dextrin. In another example, the composition comprises about 3% w / w dextrin. In another example, the composition comprises about 3.5% w / w dextrin. In a further example, the composition comprises about 4% w / w dextrin. In another example, the composition comprises about 4.5% w / w dextrin. In a further example, the composition comprises about 5% w / w dextrin.

[0195] In one example, the composition contains about 2 mg to about 50 mg of dextrin per dose.

[0196] In one example, the composition comprises at least about 2 mg of dextrin per dose.

[0197] In one example, the composition comprises at least about 4 mg of dextrin per dose.

[0198] In one example, the composition contains at least about 5 mg of dextrin per dose. In one example, the composition contains at least about 10 mg of dextrin per dose. In one example, the composition contains at least about 15 mg of dextrin per dose. In one example, the composition contains at least about 20 mg of dextrin per dose. In one example, the composition contains at least about 25 mg of dextrin per dose. In one example, the composition contains at least about 30 mg of dextrin per dose. In one example, the composition contains at least about 35 mg of dextrin per dose. In one example, the composition contains at least about 40 mg of dextrin per dose. In one example, the composition contains at least about 45 mg of dextrin per dose.

[0199] In one example, the composition contains about 2 mg of dextrin per dose.

[0200] In one example, the composition contains about 4 mg of dextrin per dose.

[0201] In one example, the composition contains about 10 mg of dextrin per dose. In one example, the composition contains about 15 mg of dextrin per dose. In one example, the composition contains about 20 mg of dextrin per dose. In one example, the composition contains about 25 mg of dextrin per dose. In one example, the composition contains about 30 mg of dextrin per dose. In one example, the composition contains about 35 mg of dextrin per dose. In one example, the composition contains about 40 mg of dextrin per dose. In one example, the composition contains about 45 mg of dextrin per dose.

[0202] In one example, a composition used in accordance with the invention described herein (e.g., a composition used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or a composition used in a method of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) comprises L-cysteine and dextrin. The L-cysteine and dextrin can be present in any suitable concentration in the compositions described herein. Suitable concentrations or amounts of L-cysteine and dextrin are described elsewhere herein and apply equally to compositions described herein that include both L-cysteine and dextrin. Illustrative examples of suitable concentrations or amounts are provided below.

[0203] In one example, the composition contains about 0.5% to 5% dextrin and about 10% to 40% L-cysteine. In each of these examples, the composition may also contain about 50% to 90% rice bran (e.g., fermented rice bran) and / or about 10,000 to 15,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0204] In one example, the compositions described herein contain at least about 0.5% w / w dextrin and at least about 10% w / w L-cysteine. For example, the compositions described herein may contain about 0.5% w / w dextrin and about 10% w / w L-cysteine. In each of these examples, the composition may also contain at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran) and / or at least 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 67% w / w rice bran (e.g., about 67% w / w fermented rice bran) and / or 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0205] In one example, the compositions described herein comprise at least about 0.5% w / w dextrin and at least about 20% w / w L-cysteine. For example, the compositions described herein may comprise about 0.5% w / w dextrin and about 20% w / w L-cysteine. In each of these examples, the composition may also comprise at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran) and / or at least 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may comprise about 67% w / w rice bran (e.g., about 67% w / w fermented rice bran) and / or 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0206] In one example, the compositions described herein comprise at least about 0.5% w / w dextrin and at least about 20% w / w L-cysteine. For example, the compositions described herein may comprise about 0.5% w / w dextrin and about 20% w / w L-cysteine. In each of these examples, the composition may also comprise at least about 73% w / w rice bran (e.g., at least about 73% w / w fermented rice bran) and / or at least 100,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may comprise about 73% w / w rice bran (e.g., about 73% w / w fermented rice bran) and / or 100,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0207] In one example, the compositions described herein comprise at least about 0.5% w / w dextrin and at least about 30% w / w L-cysteine. For example, the compositions described herein may comprise about 0.5% w / w dextrin and about 30% w / w L-cysteine. In each of these examples, the composition may also comprise at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran) and / or at least 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may comprise about 67% w / w rice bran (e.g., about 67% w / w fermented rice bran) and / or 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0208] In one example, the compositions described herein comprise at least about 5% w / w dextrin and at least about 20% w / w L-cysteine. For example, the compositions described herein may comprise about 5% w / w dextrin and about 20% w / w L-cysteine. In each of these examples, the composition may also comprise at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran) and / or at least 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may comprise about 67% w / w rice bran (e.g., about 67% w / w fermented rice bran) and / or 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0209] In one example, the compositions described herein comprise at least about 3% w / w dextrin and at least about 30% w / w L-cysteine. For example, the compositions described herein may comprise about 3% w / w dextrin and about 30% w / w L-cysteine. In each of these examples, the composition may also comprise at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran) and / or at least 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may comprise about 67% w / w rice bran (e.g., about 67% w / w fermented rice bran) and / or 10,000 cfu / g Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0210] In one example, the composition contains about 2 mg to about 50 mg of dextrin per dose and about 38 mg to about 200 mg of L-cysteine per dose. In each of these examples, the composition contains about 300 mg to about 600 mg of rice bran per dose (e.g., fermented rice bran per dose) and / or about 5,000 cfu of Bacillus bacteria per dose to about 1 x 108 It may also include cfu / dosage of Bacillus bacteria (eg, B. subtilis and / or B. coagulans).

[0211] In one example, the compositions described herein contain at least about 2 mg of dextrin per dose and at least about 38 mg of L-cysteine per dose. For example, the compositions described herein may contain about 2 mg of dextrin per dose and about 38 mg of L-cysteine per dose. In each of these examples, the composition may also contain at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose) and / or at least 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 300 mg of rice bran per dose (e.g., about 300 mg of fermented rice bran per dose) and / or 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0212] In one example, the compositions described herein contain at least about 2 mg of dextrin and at least about 76 mg of L-cysteine per dose. For example, the compositions described herein may contain about 2 mg of dextrin per dose and about 76 mg of L-cysteine per dose. In each of these examples, the composition may also contain at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose) and / or at least 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 300 mg of rice bran per dose (e.g., about 300 mg of fermented rice bran per dose) and / or 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0213] In one example, the compositions described herein contain at least about 2 mg of dextrin per dose and at least about 114 mg of L-cysteine per dose. For example, the compositions described herein may contain about 2 mg of dextrin per dose and about 114 mg of L-cysteine per dose. In each of these examples, the composition may also contain at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose) and / or at least 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 300 mg of rice bran per dose (e.g., about 300 mg of fermented rice bran per dose) and / or 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0214] In one example, the compositions described herein contain at least about 20 mg of dextrin per dose and at least about 76 mg of L-cysteine per dose. For example, the compositions described herein may contain about 20 mg of dextrin per dose and about 76 mg of L-cysteine per dose. In each of these examples, the composition may also contain at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose) and / or at least 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 300 mg of rice bran per dose (e.g., about 300 mg of fermented rice bran per dose) and / or 5,000 cfu of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0215] In one example, the compositions described herein contain at least about 4 mg of dextrin per dose and at least about 150 mg of L-cysteine per dose. For example, the compositions described herein may contain about 4 mg of dextrin per dose and about 150 mg of L-cysteine per dose. In each of these examples, the compositions contain at least about 552 mg of rice bran per dose (e.g., at least about 552 mg of fermented rice bran per dose) and / or at least 1×10 5 cfu of Bacillus bacteria / dose (e.g., B. subtilis and / or B. coagulans). For example, in each of these examples, the composition may contain about 552 mg of rice bran / dose (e.g., about 552 mg of fermented rice bran / dose) and / or 1 x 10 5 cfu / dose of Bacillus bacteria (eg, B. subtilis and / or B. coagulans).

[0216] As discussed elsewhere herein, it is suggested that AB001 has four actions that control glucose. In one example, without being bound by theory, it is suggested that when Bacillus cells and their endospores are resuscitated in the duodenum and small intestine, the cells scan the biochemical conditions of their microenvironment and begin secreting a unique set of bioactive substances that optimize conditions, e.g., pH, conductivity, and electrolytes, for their survival and growth. Nutrients and substrates are essential for survival and subsequent growth. As discussed elsewhere herein, when glucose and other carbohydrates are present in the microenvironment, carbohydrate-targeting enzymes (e.g., glucose) are secreted to advantageously degrade glucose.

[0217] As noted elsewhere herein, compositions used in accordance with the inventions described herein (e.g., compositions used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject and / or compositions used in methods of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) can include a grain (e.g., a grain grain component such as rice bran). A grain is any herb grown (cultivated) for the edible portion of its grain (a type of grain botanically called spikelets), which is composed of the endosperm, germ, and bran layer. The term grain can also refer to the resulting grain (specifically, the "grain") itself. Cereal grains are seeds from herbs such as wheat, millet, rice, barley, oats, rye, triticale, sorghum, and maize (corn).

[0218] In some examples, compositions used in accordance with the present invention may include a grain (e.g., cereal grain) selected from the group consisting of wheat, millet, rice, barley, oats, rye, triticale, sorghum, and maize (corn). Those of skill in the art will be able to readily identify grains (e.g., cereal grains) suitable for use in the compositions described herein.

[0219] Bran, also known as miller's bran, is the tough outer layer of cereal grains. Bran includes aleurone and the pericarp. Corn bran also includes the rachis (cap). Along with the germ, bran is an integral part of whole grains and is often produced as a by-product of flour milling in the production of refined grains. Bran is present in cereal grains, including rice, corn, wheat, oats, barley, rye, and millet. Thus, in some instances, compositions used in accordance with the present invention may include cereal bran. In some instances, compositions used in accordance with the present invention may include cereal bran selected from the group consisting of rice bran, corn bran, wheat bran, oat bran, barley bran, rye bran, and millet bran.

[0220] In some examples, compositions used in accordance with the present invention may include oat bran.

[0221] In some examples, compositions used in accordance with the present invention may include rice bran.

[0222] Rice bran is a by-product of the rice milling process. Generally, milling produces approximately 15% w / w broken grains, approximately 10% w / w rice bran, approximately 20% rice husk, and approximately 55% w / w whole grains. Rice bran's composition (weight percent) is approximately 11-13% water, 18-21% crude oil, 14-16% crude protein, 8-10% crude fiber, 9-12% ash, and 33-36% carbohydrates. Rice bran has a rancid odor and taste due to the presence of natural lipases that hydrolyze oil into glycerol and free fatty acids. As used herein, "rice bran" refers to the tough outer layer of rice, including the aleurone and pericarp. Along with the germ, rice bran is an integral part of the whole grain and, as noted above, is often produced as a by-product of rice milling in the production of refined rice.

[0223] Compositions used in accordance with the invention described herein may contain rice bran in any suitable form. Suitable forms include raw rice bran, freshly milled (unhydrolyzed) full-fat rice bran, reduced-fat rice bran, defatted rice bran, fermented rice bran, stabilized rice bran, and the like. Raw rice bran is the rice bran obtained after rice milling. Reduced-fat rice bran and defatted rice bran are obtained from full-fat rice bran, such as by solvent extraction. Full-fat rice bran has a fat content of about 14-18% by weight, while reduced-fat rice bran and defatted rice bran have about 3-14% and less than 3% fat, respectively, by weight.

[0224] In one example, the rice bran is formulated as raw rice bran, freshly milled (unhydrolyzed) full-fat rice bran, reduced-fat rice bran, defatted rice bran, fermented rice bran, and / or stabilized rice bran.

[0225] In another more preferred example, the rice bran is formulated as fermented rice bran and / or stabilized rice bran.

[0226] In a further example, the rice bran is fermented rice bran, which undergoes a natural / spontaneous fermentation process by naturally occurring microbial strains, typically soil strains, such as Bacillus and Pediococcus.

[0227] As used herein, "fermented rice bran" refers to rice bran that has undergone a fermentation process. Fermented rice bran contains probiotic microorganisms that stabilize the small intestine microbiome.

[0228] Generally, the terms "microbiota" and "microbiome" are used to describe the microbial flora.

[0229] As used herein, "stabilized rice bran" refers to rice bran that has been heated for a short period of time, for example, by passing it through a high-temperature, high-pressure extruder. This heat stabilizes the rice bran. In other words, "stabilized rice bran" is therefore heat-treated rice bran. For example, rice bran can be stabilized by heating it at 130°C for less than 10 seconds after milling. In some examples, stabilized rice bran is a dietary fiber. Typically, stabilized rice bran is a dietary fiber that can be catabolized in the colon. As a dietary fiber, stabilized rice bran is a prebiotic that interacts with the colonic microbiome.

[0230] The microbial components of fermented rice bran may have probiotic and stabilizing effects on the small intestinal microbiome, and the dietary fiber in rice bran may stabilize colonic conditions. Another mechanism of action may be the reduction of intestinal oxidative stress, which normalizes the barrier function of the intestinal mucosa.

[0231] Rice bran (e.g., fermented rice bran) is present in the compositions described herein in a suitable amount or concentration, as described below. The amount or concentration of rice bran may therefore be described by reference to the % w / w of the composition or by the total weight (e.g., mg) per dose of the composition (in other words, the weight per effective dose). As will be apparent to one of skill in the art, and as described in more detail elsewhere herein, a dose may include one or more dosage units (e.g., two dosage units). In instances where multiple dosage units are used to provide an effective dose, the weight per dose corresponds to the total weight of rice bran across all the multiple dosage units.

[0232] In one example, the composition comprises about 50% w / w to about 90% w / w rice bran.

[0233] In another example, the composition comprises about 60% w / w to about 80% w / w rice bran, hi another example, the composition comprises about 73% w / w to about 79% w / w rice bran.

[0234] In one example, the composition comprises at least about 50% w / w rice bran. In another example, the composition comprises at least about 55% w / w rice bran. In one example, the composition comprises at least about 60% w / w rice bran. In a further example, the composition comprises at least about 65% w / w rice bran. In one example, the composition comprises at least about 73% w / w rice bran. In one example, the composition comprises at least about 75% w / w rice bran. In one example, the composition comprises at least about 79% w / w rice bran. In a further example, the composition comprises at least about 85% w / w rice bran. In one example, the composition comprises at least about 90% w / w rice bran.

[0235] In one example, the composition comprises about 50% w / w rice bran. In another example, the composition comprises about 55% w / w rice bran. In one example, the composition comprises about 60% w / w rice bran. In a further example, the composition comprises about 65% w / w rice bran. In one example, the composition comprises about 73% w / w rice bran. In one example, the composition comprises about 75% w / w rice bran. In one example, the composition comprises about 79% w / w rice bran. In a further example, the composition comprises about 85% w / w rice bran. In one example, the composition comprises about 90% w / w rice bran.

[0236] In particular examples, the composition comprises at least about 67% w / w rice bran (e.g., fermented rice bran). For example, the composition may comprise about 67% w / w rice bran (e.g., fermented rice).

[0237] In particular examples, the composition comprises at least about 73% w / w rice bran (e.g., fermented rice). For example, the composition may comprise about 73% w / w rice bran (e.g., fermented rice).

[0238] In another example, the composition comprises at least about 79% w / w rice bran (e.g., fermented rice). For example, the composition may comprise about 79% w / w rice bran (e.g., fermented rice).

[0239] In another example, the composition comprises at least about 79.5% w / w rice bran (e.g., fermented rice). For example, the composition may comprise about 79.5% w / w rice bran (e.g., fermented rice).

[0240] In one example, the composition includes at least about 300 mg of rice bran (e.g., fermented rice bran) per dose. As described elsewhere herein, a dose may be formulated as two capsules.

[0241] In one example, the composition contains at least about 300 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains at least about 350 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains at least about 400 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains at least about 450 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains at least about 500 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains at least about 550 mg (e.g., at least about 552 mg) of rice bran (e.g., fermented rice bran) per dose.

[0242] In one example, the composition contains about 300 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains about 350 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains about 400 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains about 450 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains about 500 mg of rice bran (e.g., fermented rice bran) per dose. In one example, the composition contains about 550 mg (e.g., about 552 mg) of rice bran (e.g., fermented rice bran) per dose.

[0243] As discussed elsewhere herein, compositions comprising one or more Bacillus species (e.g., B. subtilis and / or B. coagulans), rice bran (e.g., fermented rice bran), L-cysteine, and a high molecular weight, low-osmolality carbohydrate (e.g., dextrin) are provided for use in accordance with the present invention (e.g., for the treatment and / or prevention of type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or in methods of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject). The one or more Bacillus species (e.g., B. subtilis and / or B. coagulans), rice bran (e.g., fermented rice bran), L-cysteine, and a high molecular weight, low-osmolality carbohydrate (e.g., dextrin) can be present in any suitable amount or concentration in a composition for use in accordance with the present invention. Suitable amounts and concentrations of one or more Bacillus species (e.g., B. subtilis and / or B. coagulans), rice bran (e.g., fermented rice bran), L-cysteine, and high molecular weight, low osmolality carbohydrates (e.g., dextrin) are described elsewhere herein. Illustrative examples of suitable concentrations are provided below.

[0244] In one example, the composition comprises about 0.5% w / w to 5% w / w dextrin, about 10% w / w to about 40% w / w L-cysteine, about 50% w / w to about 90% w / w rice bran (e.g., fermented rice bran), and about 10,000 cfu / g to about 15,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0245] In one example, a composition described herein comprises at least about 0.5% w / w dextrin, at least about 10% w / w L-cysteine, at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran), and at least 10,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0246] In one example, a composition described herein comprises at least about 0.5% w / w dextrin, at least about 20% w / w L-cysteine, at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran), and at least 10,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0247] In one example, a composition described herein comprises at least about 0.5% w / w dextrin, at least about 20% w / w L-cysteine, at least about 73% w / w rice bran (e.g., at least about 73% w / w fermented rice bran), and at least 100,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0248] In one example, a composition described herein comprises at least about 0.5% w / w dextrin, at least about 30% w / w L-cysteine, at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran), and at least 10,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0249] In one example, a composition described herein comprises at least about 5% w / w dextrin, at least about 20% w / w L-cysteine, at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran), and at least 10,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0250] In one example, a composition described herein comprises at least about 3% w / w dextrin, at least about 30% w / w L-cysteine, at least about 67% w / w rice bran (e.g., at least about 67% w / w fermented rice bran), and at least 10,000 cfu / g of Bacillus bacteria (e.g., B. subtilis and / or B. coagulans).

[0251] In one example, the composition comprises about 2 mg to about 50 mg of dextrin per dose, about 38 mg to about 200 mg of L-cysteine per dose, about 300 mg to about 600 mg of rice bran per dose (e.g., fermented rice bran per dose), and about 5,000 cfu of Bacillus bacteria per dose to about 1×10 8 Contains cfu of Bacillus bacteria / dose (e.g., B. subtilis and / or B. coagulans).

[0252] In one example, a composition described herein includes at least about 2 mg of dextrin per dose, at least about 38 mg of L-cysteine per dose, at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose), and at least 5,000 cfu of Bacillus bacteria per dose (e.g., B. subtilis and / or B. coagulans).

[0253] In one example, a composition described herein includes at least about 2 mg of dextrin, at least about 76 mg of L-cysteine per dose, at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose), and at least 5,000 cfu of Bacillus bacteria per dose (e.g., B. subtilis and / or B. coagulans).

[0254] In one example, a composition described herein includes at least about 2 mg of dextrin per dose, at least about 114 mg of L-cysteine per dose, at least about 300 mg of rice bran per dose (e.g., at least about 300 mg of fermented rice bran per dose), and at least 5,000 cfu of Bacillus bacteria per dose (e.g., B. subtilis and / or B. coagulans).

[0255] In one example, a composition described herein includes at least about 20 mg dextrin per dose, at least about 76 mg L-cysteine per dose, at least about 300 mg rice bran per dose (e.g., at least about 300 mg fermented rice bran per dose), and at least 5,000 cfu of Bacillus bacteria per dose (e.g., B. subtilis and / or B. coagulans).

[0256] In one example, the compositions described herein comprise at least about 4 mg dextrin / dose, at least about 150 mg L-cysteine / dose, at least about 552 mg rice bran / dose (e.g., at least about 552 mg fermented rice bran / dose), and at least 1×10 5 Contains cfu of Bacillus bacteria / dose (e.g., B. subtilis and / or B. coagulans).

[0257] In certain examples, the compositions described herein comprise at least about 0.5% w / w dextrin, at least about 20% w / w L-cysteine, and at least about 73% w / w rice bran (e.g., at least about 73% w / w fermented rice bran). For example, the compositions described herein may comprise about 0.5% w / w dextrin, about 20% w / w L-cysteine, and about 73% w / w rice bran (e.g., about 73% w / w fermented rice bran). In these examples, the fermented rice bran may comprise one or more Bacillus species.

[0258] In a particular example, the compositions described herein comprise about 79.5% w / w fermented rice bran (comprising one or more Bacillus species), about 0.5% w / w dextrin, and about 20.0% w / w L-cysteine.

[0259] Compositions used in accordance with the invention described herein (e.g., compositions used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or compositions used in methods of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) can include one or more additional components, e.g., one or more additional ingredients. For example, the compositions described herein can further include an emulsifier, a filler, an excipient, and / or an inactive ingredient. One of ordinary skill in the art will readily understand what is meant by "emulsifier," "filler," "excipient," and "inactive ingredient" in the context of the compositions used in accordance with the invention provided herein. Non-limiting examples of additional components include vitamin B12, magnesium salts of fatty acids (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose).

[0260] The additional components described herein may be in any suitable form, which will be readily identifiable by one skilled in the art.

[0261] Thus, in one example, the compositions described herein may further comprise one or more of vitamin B12, a fatty acid magnesium salt (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose). In another example, the compositions described herein may further comprise two or more of vitamin B12, a fatty acid magnesium salt (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose). In another example, the compositions described herein may further comprise three or more of vitamin B12, a fatty acid magnesium salt (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose). In another example, the compositions described herein may further comprise four or more of vitamin B12, a fatty acid magnesium salt (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose).

[0262] In one example, the described composition further comprises vitamin B12, a magnesium salt of a fatty acid (e.g., magnesium stearate), calcium phosphate, potassium phosphate, silicon dioxide, and cellulose (e.g., microcrystalline cellulose).

[0263] An example of a fatty acid magnesium salt is magnesium stearate. Thus, in one example, the composition described herein may further comprise magnesium stearate. Magnesium stearate is a magnesium salt of stearic acid, a fatty acid. Magnesium salts of fatty acids (e.g., magnesium stearate) can be used as excipients, inactive ingredients, and / or lubricants in machine manufacturing. Magnesium stearate is a GRAS listed ingredient.

[0264] In some instances, vitamin B12 is added for regulatory purposes.

[0265] In some instances, the magnesium salts (e.g., magnesium stearate), calcium salts, and / or potassium salts are inactive ingredients (e.g., they have no clinical effect). For example, the magnesium salts (e.g., magnesium stearate), calcium salts, and / or potassium salts may be added as filling aids (e.g., fillers) to help capsule filling machines operate efficiently.

[0266] In some instances, a chromium salt is added for conditioning purposes. For example, the chromium salt can be chromium chloride. Any suitable amount of chromium salt (e.g., chromium chloride) can be used.

[0267] In one example, the compositions described herein comprise at least about 20 mcg of chromium per dose. In one example, the compositions described herein comprise about 20 mcg of chromium per dose.

[0268] In one example, the compositions described herein contain about 80 mcg to about 100 mcg of CrCl3 (chromium chloride) per dose.

[0269] In one example, a composition used in accordance with the invention described herein (e.g., a composition used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or a composition used in a method of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject) further comprises microcrystalline cellulose. Microcrystalline cellulose may be used as an emulsifier, a filling aid, and / or an inactive ingredient. Microcrystalline cellulose (and maltodextrin as well) may be used as an excipient to form a solid mass during tablet compression and as a fluidity agent in making capsule formulations. In some examples, the compositions described herein further comprise maltodextrin.

[0270] In one example, the composition comprises about 5% w / w to about 50% w / w microcrystalline cellulose. In another example, the composition comprises about 5% w / w to about 35% w / w microcrystalline cellulose. In another example, the composition comprises about 5% w / w to about 20% w / w microcrystalline cellulose. In another example, the composition comprises about 5% w / w to about 15% w / w microcrystalline cellulose. In another example, the composition comprises about 5% w / w to about 8% w / w microcrystalline cellulose.

[0271] In one example, the composition comprises at least about 5% w / w microcrystalline cellulose.

[0272] In one example, the composition comprises at least about 8% w / w microcrystalline cellulose.

[0273] In another example, the composition comprises at least about 10% w / w microcrystalline cellulose. In another example, the composition comprises at least about 15% w / w microcrystalline cellulose. In a further example, the composition comprises at least about 20% w / w microcrystalline cellulose. In another example, the composition comprises at least about 25% w / w microcrystalline cellulose. In another example, the composition comprises at least about 30% w / w microcrystalline cellulose. In a further example, the composition comprises at least about 35% w / w microcrystalline cellulose. In another example, the composition comprises at least about 40% w / w microcrystalline cellulose. In another example, the composition comprises at least about 45% w / w microcrystalline cellulose. In another example, the composition comprises at least about 50% w / w microcrystalline cellulose.

[0274] In one example, the composition comprises about 5% w / w microcrystalline cellulose.

[0275] In one example, the composition comprises about 8% w / w microcrystalline cellulose.

[0276] In another example, the composition comprises about 10% w / w microcrystalline cellulose. In another example, the composition comprises about 15% w / w microcrystalline cellulose. In a further example, the composition comprises about 20% w / w microcrystalline cellulose. In another example, the composition comprises about 25% w / w microcrystalline cellulose. In a further example, the composition comprises about 30% w / w microcrystalline cellulose. In another example, the composition comprises about 35% w / w microcrystalline cellulose. In another example, the composition comprises about 40% w / w microcrystalline cellulose. In another example, the composition comprises about 45% w / w microcrystalline cellulose. In another example, the composition comprises about 50% w / w microcrystalline cellulose.

[0277] In one example, the compositions described herein contain at least about 30 mg of microcrystalline cellulose per dose. As described elsewhere herein, a dose may be formulated in two capsules.

[0278] In one example, the compositions described herein comprise at least about 40 mg of microcrystalline cellulose per dose.

[0279] In one example, the compositions described herein contain at least about 50 mg of microcrystalline cellulose per dose. In one example, the compositions described herein contain at least about 60 mg of microcrystalline cellulose per dose. In one example, the compositions described herein contain at least about 70 mg of microcrystalline cellulose per dose. In one example, the compositions described herein contain at least about 80 mg of microcrystalline cellulose per dose. In one example, the compositions described herein contain at least about 90 mg of microcrystalline cellulose per dose. In one example, the compositions described herein contain at least about 100 mg of microcrystalline cellulose per dose.

[0280] In one example, a composition used in accordance with the invention described herein (e.g., a composition used to treat and / or prevent type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject, and / or a composition in a method of treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof) further comprises magnesium stearate, which may be used as an emulsifier, a loading aid, and / or an inactive ingredient.

[0281] In one example, the composition comprises about 0.2% w / w to about 1.5% w / w magnesium stearate. In another example, the composition comprises about 0.5% w / w to about 1.4% w / w magnesium stearate. In another example, the composition comprises about 0.6% w / w to about 1.3% w / w magnesium stearate.

[0282] In one example, the composition comprises at least about 0.2% w / w magnesium stearate. In another example, the composition comprises at least about 0.3% w / w magnesium stearate. In another example, the composition comprises at least about 0.4% w / w magnesium stearate. In a further example, the composition comprises at least about 0.5% w / w magnesium stearate.

[0283] In another example, the composition comprises at least about 0.6% w / w magnesium stearate.

[0284] In another example, the composition comprises at least about 0.7% w / w magnesium stearate. In a further example, the composition comprises at least about 0.8% w / w magnesium stearate. In another example, the composition comprises at least about 0.9% w / w magnesium stearate.

[0285] In another example, the composition comprises at least about 1.0% w / w magnesium stearate.

[0286] In another example, the composition comprises at least about 1.1% w / w magnesium stearate. In another example, the composition comprises at least about 1.2% w / w magnesium stearate. In another example, the composition comprises at least about 1.3% w / w magnesium stearate. In another example, the composition comprises at least about 1.4% w / w magnesium stearate. In another example, the composition comprises at least about 1.5% w / w magnesium stearate.

[0287] In one example, the composition comprises about 0.2% w / w magnesium stearate. In another example, the composition comprises about 0.3% w / w magnesium stearate. In another example, the composition comprises about 0.4% w / w magnesium stearate. In a further example, the composition comprises about 0.5% w / w magnesium stearate.

[0288] In another example, the composition comprises about 0.6% w / w magnesium stearate.

[0289] In another example, the composition comprises about 0.7% w / w magnesium stearate. In a further example, the composition comprises about 0.8% w / w magnesium stearate. In another example, the composition comprises about 0.9% w / w magnesium stearate.

[0290] In another example, the composition comprises about 1.0% w / w magnesium stearate.

[0291] In another example, the composition comprises about 1.1% w / w magnesium stearate. In another example, the composition comprises about 1.2% w / w magnesium stearate. In another example, the composition comprises about 1.3% w / w magnesium stearate. In another example, the composition comprises about 1.4% w / w magnesium stearate. In another example, the composition comprises about 1.5% w / w magnesium stearate.

[0292] In one example, the compositions described herein contain at least about 4 mg of magnesium stearate per dose, which may be formulated as two capsules, as described elsewhere herein.

[0293] In one example, the compositions described herein contain at least about 5 mg of magnesium stearate per dose.

[0294] In one example, the compositions described herein contain at least about 6 mg of magnesium stearate per dose. In one example, the compositions described herein contain at least about 7 mg of magnesium stearate per dose. In one example, the compositions described herein contain at least about 8 mg of magnesium stearate per dose. In one example, the compositions described herein contain at least about 9 mg of magnesium stearate per dose. In one example, the compositions described herein contain at least about 10 mg of magnesium stearate per dose.

[0295] In one example, a composition used in accordance with the invention described herein (e.g., a composition used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or a composition in a method of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof) further comprises silicon dioxide, which may be used as an emulsifier, a loading aid, and / or an inactive ingredient.

[0296] In one example, the composition comprises about 0.5% to about 4% w / w silicon dioxide. In another example, the composition comprises about 0.7% to about 3% w / w silicon dioxide. In another example, the composition comprises about 0.9% to about 2% w / w silicon dioxide.

[0297] In one example, the composition comprises at least about 0.5% w / w silicon dioxide.

[0298] In another example, the composition comprises at least about 1% w / w silicon dioxide.

[0299] In another example, the composition comprises at least about 1.5% w / w silicon dioxide. In a further example, the composition comprises at least about 2% w / w silicon dioxide. In another example, the composition comprises at least about 2.5% w / w silicon dioxide. In another example, the composition comprises at least about 3% w / w silicon dioxide. In a further example, the composition comprises at least about 3.5% w / w silicon dioxide. In another example, the composition comprises at least about 4% w / w silicon dioxide.

[0300] In one example, the composition comprises about 0.5% w / w silicon dioxide.

[0301] In another example, the composition comprises about 1% w / w silicon dioxide.

[0302] In another example, the composition comprises about 1.5% w / w silicon dioxide. In a further example, the composition comprises about 2% w / w silicon dioxide. In another example, the composition comprises about 2.5% w / w silicon dioxide. In another example, the composition comprises about 3% w / w silicon dioxide. In a further example, the composition comprises about 3.5% w / w silicon dioxide. In another example, the composition comprises about 4% w / w silicon dioxide.

[0303] In one example, the compositions described herein contain at least about 4 mg of silicon dioxide per dose. As described elsewhere herein, one dose may be formulated in two capsules.

[0304] In one example, the compositions described herein comprise at least about 5 mg of silicon dioxide per dose.In one example, the compositions described herein comprise at least about 6 mg of silicon dioxide per dose.In one example, the compositions described herein comprise at least about 7 mg of silicon dioxide per dose.In one example, the compositions described herein comprise at least about 8 mg of silicon dioxide per dose.

[0305] In one example, a composition used in accordance with the invention described herein (e.g., a composition used to treat and / or prevent type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, and / or a composition in a method of treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof) further comprises vitamin B12. Vitamin B12 may be included for regulatory purposes in the compositions described herein. In one example, the composition comprises at least 15% of the Required Daily Intake (RDI), i.e., 0.38 mcg to a maximum of 2.4 mcg in the EU and 2.5 mcg in the US.

[0306] In one example, the compositions described herein contain at least about 0.76 mcg of vitamin B12 per dose. In one example, the compositions described herein contain at least about 0.9 mcg of vitamin B12 per dose.

[0307] The composition can be formulated in any suitable form. For example, the composition can be in the form of a tablet or capsule. In one example, the composition is formulated as an acid-resistant tablet or capsule. Generally, "capsule" refers to both an empty capsule and a filled capsule, while "shell" refers specifically to an empty capsule unless the context requires otherwise.

[0308] In some examples, the compositions described herein are contained within a capsule (e.g., within a shell). In some examples, the compositions described herein are contained within an acid-resistant capsule (e.g., within an acid-resistant shell). As will be apparent to one of skill in the art, references to the % w / w of an ingredient present in a composition do not take into account any weight that may be of the capsule (and therefore only take into account the % w / w of the composition within the capsule).

[0309] In one example, an acid-resistant tablet or capsule comprises a film coating, and the film coating comprises hydroxypropylmethylcellulose (HPMC). HPMC is a semi-synthetic, inert, viscoelastic polymer used in various applications. For example, HPMC can be used as an excipient in oral formulations of tablets and capsules, and some grades can function as a controlled-release agent to delay the release of pharmaceutical compounds in the gastrointestinal tract. In tablets, HPMC can also be used as a binder and / or as a component of tablet coating.

[0310] In one example, the composition used in accordance with the invention described herein is contained within a shell (of a capsule), and the shell comprises hydroxypropylmethylcellulose (HPMC).

[0311] Those of skill in the art will understand the meaning of "acid-resistant" in the context of the present invention, and particularly in the context of the ingestible compositions described herein. For example, the compositions described herein may be formulated as acid-resistant tablets or capsules or contained within an acid-resistant capsule (e.g., an acid-resistant shell) that dissolves upon reaching the duodenum, releasing microbial cells and spores and allowing them to settle in the upper intestinal tract.

[0312] In one example, the composition is present in a capsule, and the capsule contains about 80 mg to about 100 mg of HPMC. For example, the capsule may contain at least about 80 mg of HPMC. In one example, the capsule contains at least about 90 mg of HPMC. In one example, the capsule contains at least about 100 mg of HPMC. In one example, the capsule contains about 80 mg of HPMC. In one example, the capsule contains about 90 mg of HPMC. In another example, the capsule contains about 100 mg of HPMC.

[0313] Typically, HPMC is present as a film coating on the outer surface of the capsules described herein.

[0314] The compositions described herein can be in unit dosage form.When the compositions described herein are in unit dosage form, one tablet or capsule can be administered, which constitutes one dose, or two tablets or capsules can be administered, which constitutes one dose.Suitable dosage and administration schedule can be determined by those skilled in the art based on the following examples.Therefore, in one example, one dose of the composition according to the present invention comprises multiple (for example, two) small tablets or capsules.

[0315] The term "dosage form," as used herein, refers to an amount of medication taken at one time, optionally at regular intervals. Herein, this is also referred to as a "dose."

[0316] In one example, the present invention provides a solid unit dosage form for oral administration.

[0317] In a specific example, a dose of the composition described herein contains about 300 mg of fermented rice bran, about 38 mg of L-cysteine, about 30 mg of microcrystalline cellulose, about 4 mg of magnesium stearate, about 4 mg of silicon dioxide, about 2 mg of dextrin, and about 5,000 cfu of Bacillus bacteria. Optionally, a dose of the composition described herein may further contain about 0.76 μg (mcg) of vitamin B12 and / or any suitable amount of chromium chloride. In this specific example, the dose may be formulated as two tablets or capsules. In other words, the amount of a component may represent the total amount of the component present in the dose (i.e., in two tablets or capsules). Therefore, those skilled in the art will understand that at least each of these components, in at least these amounts, provides an effective dose of the present invention. Thus, in one example, a dose of the compositions described herein comprises at least about 300 mg of fermented rice bran, at least about 38 mg of L-cysteine, at least about 30 mg of microcrystalline cellulose, at least about 4 mg of magnesium stearate, at least about 4 mg of silicon dioxide, at least about 2 mg of dextrin, and at least about 5,000 cfu of Bacillus bacteria. Optionally, a dose of the compositions described herein may further comprise at least about 0.76 mcg of vitamin B12 and / or any suitable amount of chromium chloride.

[0318] In a specific example, a dose of the composition described herein contains about 552 mg of fermented rice bran, about 150 mg of L-cysteine, about 40 mg of microcrystalline cellulose, about 4.8 mg of magnesium stearate, about 4 mg of silicon dioxide, about 4 mg of dextrin, and about 100,000 cfu of Bacillus bacteria. Optionally, a dose of the composition described herein may further contain about 0.9 μg (mcg) of vitamin B12 and / or any suitable amount of chromium chloride. In this specific example, the dose may be formulated as two tablets or capsules. In other words, the amount of a component may represent the total amount of the component present in the dose (i.e., in two tablets or capsules). Therefore, those skilled in the art will understand that at least each of these components, in at least these amounts, provides an effective dose of the present invention. Thus, in one example, a dose of the compositions described herein comprises at least about 552 mg of fermented rice bran, at least about 150 mg of L-cysteine, at least about 40 mg of microcrystalline cellulose, at least about 4.8 mg of magnesium stearate, at least about 4 mg of silicon dioxide, at least about 4 mg of dextrin, and at least about 100,000 cfu of Bacillus bacteria. Optionally, a dose of the compositions described herein may further comprise at least about 0.9 mcg of vitamin B12 and / or any suitable amount of chromium chloride.

[0319] Typically, HPMC is present as a film coating on the outer surface of the capsules described herein.

[0320] The compositions described herein may be used as (or used as part of) a dietary supplement, nutraceutical, food composition, medical food, or pharmaceutical product.

[0321] The term "dietary supplement" or "food supplement," as used herein, refers to a composition taken in addition to or between meals.

[0322] The term "food composition," as used herein, refers to any type of composition that can be eaten and / or drunk without causing toxic symptoms in the subject eating or drinking the composition.

[0323] The use of the composition described herein for decomposing glucose is provided herein.Accordingly, in one example, the use of a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for decomposing glucose is provided herein.The term "decomposing glucose" is described elsewhere herein and is equally applicable here.

[0324] The composition can be used to degrade glucose in a subject (e.g., the use can be in vivo). The subject can be any suitable subject, for example, the subject can be a human. The subject can be a human who is about to ingest or has ingested carbohydrates (e.g., glucose).

[0325] In one example, the composition can be used to metabolize glucose in the digestive tract of a subject. The term "metabolizing glucose" is described elsewhere herein and applies equally here. In one example, the composition can be used to metabolize glucose in the intestine of a subject. In a particular example, the composition can be used to metabolize glucose in the small intestine of a subject. More specifically, the composition can be used to metabolize glucose in the duodenum of a subject.

[0326] In one example, the composition can be used to reduce glucose absorption into the blood of a subject. A person skilled in the art can use methods known in the art to determine the reduction in glucose absorption into the blood in the presence of the compositions provided herein (compared to when the composition is not used).

[0327] When the composition is used to decompose glucose in a subject, it can be used to reduce the blood glucose concentration in the subject.Those skilled in the art can use methods known in the art to determine the reduction in blood glucose concentration in the presence of the composition provided herein (compared to when the composition is not used).

[0328] Use of a composition described herein (e.g., a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate) for controlling blood glucose in a subject is also provided.

[0329] (a) to reduce one or more levels of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, and / or hsCRP in the blood of a subject; (b) to reduce the HOMA-IR score in a subject; and / or (c) increasing blood adiponectin levels in a subject; Uses of the compositions described herein (e.g., compositions comprising one or more Bacillus species, rice bran, L-cysteine, and high molecular weight, low osmolality carbohydrates) are also provided herein.

[0330] "HbA1c," "glucose," e.g., fasting glucose, "insulin," "intact proinsulin," "hsCRP," "HOMA-IR score," and "adiponectin" are described elsewhere herein and apply equally here.

[0331] The subject can be any suitable subject, for example, the subject can be a human. The subject can be a human who is about to ingest or has ingested carbohydrates (e.g., glucose).

[0332] In some examples, compositions used in accordance with the present invention are in the form of a pharmaceutical formulation comprising a composition of the present invention. As will be apparent to one of skill in the art, a pharmaceutical formulation is a formulation suitable for administration to a subject (e.g., to break down glucose). The pharmaceutical formulation may be for administration to a subject to prevent and / or treat a disease, condition, or illness (e.g., to treat and / or prevent type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject, and / or for administration in a method of treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject). As used herein, a pharmaceutical formulation comprises an effective amount of a composition of the present invention.

[0333] As used herein, a "pharmaceutical composition" refers to a composition and / or its finished dosage form or formulation having pharmacological activity or other direct effect in the alleviation, treatment, or prevention of disease, and is intended for human use. Pharmaceutical compositions or preparations are typically produced under good manufacturing practices (GMP) conditions. Pharmaceutical compositions or preparations can be sterile or non-sterile. If non-sterile, such pharmaceutical compositions or preparations typically meet the microbiological specifications and standards for non-sterile pharmaceutical products set forth in the United States Pharmacopoeia (USP) or the European Pharmacopoeia (EP). Thus, the compositions described herein can be formulated as pharmaceutical compositions. In some examples, the pharmaceutical composition is non-sterile.

[0334] Methods of Treatment and Use As described elsewhere herein, the compositions described herein can be used to degrade glucose in vitro or in vivo.

[0335] The present inventors have surprisingly discovered that a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate can be advantageously used to safely improve the cardiometabolic status of a subject, particularly a subject with type 2 diabetes and / or prediabetes (as illustrated in the examples below). The composition may therefore advantageously provide a valuable supplement to existing combination treatments in subjects with type 2 diabetes and / or prediabetes.

[0336] Thus, there is provided a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject.

[0337] Also provided is a method for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, comprising administering to the subject a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

[0338] Diabetes mellitus, commonly known as diabetes, is a metabolic disorder resulting in elevated blood glucose levels due to a relative or absolute deficiency of the pancreatic hormone insulin. Insulin is secreted into the bloodstream by the pancreas in response to blood glucose levels, and one of its primary functions is to direct blood glucose into body stores, thereby controlling blood glucose levels. Type 2 diabetes mellitus, formerly known as adult-onset or non-insulin-dependent diabetes, is a chronic disease characterized by derangements in both glucose and lipid metabolism. Type 2 diabetes is characterized by insulin resistance and beta cell dysfunction, including impaired first-phase insulin release, reduced beta cell pulse mass, and insulin deficiency. In other words, in patients with type 2 diabetes, the body is resistant to the effects of insulin and / or does not produce enough insulin to maintain normal glucose levels. People with type 2 diabetes have a combination of increased insulin resistance and reduced insulin secretion, which together result in hyperglycemia, as discussed below. Dyslipidemia is also a component of the metabolic derangements that characterize type 2 diabetes.

[0339] Insulin resistance refers to a physiological state in which the blood glucose-lowering effect of the natural hormone insulin is diminished, for example, due to a reduced ability of cells to respond to insulin's action (e.g., decreased sensitivity to insulin) to promote the transport of sugar glucose from the blood to muscle and other tissues. Insulin resistance ranges from normal (insulin sensitive) to insulin resistant (IR). In some instances, insulin resistance can mean that maximal doses of the hormone are unable to induce a normal anabolic response. An inadequate response to insulin results in decreased glucose absorption (primarily in muscle and adipose tissue) and increased hepatic gluconeogenesis, both of which increase circulating blood glucose concentrations. To maintain homeostasis and prevent hyperglycemia (excessive serum glucose levels), pancreatic β cells increase insulin secretion, resulting in hyperinsulinemia (high blood insulin levels). Early in the progression to diabetes, but before the onset of type 2 diabetes, the pancreas can overcome insulin resistance and maintain normoglycemia by increasing insulin production. However, later in the progression to diabetes, compensatory insulin secretion by the pancreas fails to overcome the body's insulin resistance, and normal blood (e.g., plasma) glucose concentrations can no longer be maintained, resulting in hyperglycemia. Symptoms of hyperglycemia include polyurea (excessive urination in a given time period) and polydipsia (excessive thirst). Chronic hyperglycemia adversely affects pancreatic beta-cell function through glucose desensitization (further reduction in the body's insulin sensitivity, e.g., increased insulin resistance) and beta-cell attrition and apoptosis, thereby impairing insulin secretion. This ultimately leads to overt type 2 diabetes, characterized by fasting venous whole-blood glucose concentrations typically exceeding 7.0 mmol / L (126 mg / dL).

[0340] The terms "type 2 diabetes," "T2D," or "TIID" are well defined in the art. Similarly, the term "insulin resistance" is well defined in the art.

[0341] Type 2 diabetes is an extremely costly disease, coinciding with a significant reduction in life expectancy and requiring prolonged medical care to stave off the onset of associated short- and long-term complications. These complications include hyperinsulinemia, hyperglycemia, hypoglycemia (serum glucose <50 mg / dL), ketoacidosis, increased risk of infection, microvascular complications (e.g., retinopathy, nephropathy), neuropathic complications, and macrovascular disease, including severe atherosclerotic cardiovascular disease (CVD). Diabetes-related morbidity and mortality are primarily attributable to these complications. For example, diabetes is a leading cause of blindness and a significant cause of lower-limb amputation and kidney disease.

[0342] Symptoms of type 2 diabetes are well known to those skilled in the art and include, but are not limited to, increased thirst, frequent urination, increased hunger, unintentional weight loss, fatigue, blurred vision, slow wound healing, frequent infections, numbness or tingling in the hands or feet, and darkening of the skin, usually in the armpits and neck.

[0343] As used herein, "symptom" refers to a symptom indicative of a disease, condition, or illness. In other words, a symptom refers to a phenotype associated with a disease, condition, or illness and can be used to aid in the diagnosis of those diseases, conditions, or illnesses. In some contexts, a combination of symptoms may be required to aid in the diagnosis of a disease, condition, or illness.

[0344] "Prediabetes" refers to one or more early diabetes-related conditions, including impaired glucose utilization, abnormal or poor fasting glucose levels, impaired glucose tolerance, impaired insulin sensitivity, and insulin resistance. Prediabetes is associated with the simultaneous presence of insulin resistance and beta-cell dysfunction, and these abnormalities begin before glucose changes are detectable. Prediabetes is typically characterized by blood glucose levels that are higher than normal but below the diabetic threshold, and indicates a high likelihood of developing diabetes.

[0345] Symptoms of prediabetes include, but are not limited to, increased thirst, frequent urination, increased hunger, unintentional weight loss, fatigue, blurred vision, slow wound healing, frequent infections, numbness or tingling in the hands or feet, itching or thrush in the genital area, and darkening of the skin, usually in the armpits and neck.

[0346] Symptoms of type 2 diabetes and / or prediabetes may also include insulin resistance, beta cell dysfunction, unregulated glycemic control, chronic systemic inflammation and / or intestinal inflammation, hyperinsulinemia, hyperglycemia, hypoglycemia (serum glucose <50 mg / dL), ketoacidosis, increased risk of infection, capillary complications (i.e., retinopathy, nephropathy), neuropathic complications, and macrovascular disease such as cardiovascular disease (CVD) due to severe arteriosclerosis.

[0347] Advantageously, the compositions used according to the present invention can be used to improve the symptoms of type 2 diabetes and prediabetes as described above. The inventors have specifically determined that the composition (AB001) described herein can be used to stabilize glucose levels and / or improve glycemic control, particularly in subjects with type 2 diabetes and / or prediabetes. Advantageously, the compositions described herein can also be used to reduce insulin resistance, beta cell dysfunction, and / or inflammation (e.g., chronic systemic inflammation and / or intestinal inflammation) in subjects with type 2 diabetes and / or prediabetes. In some examples, the inflammation can be intestinal and / or pancreatic inflammation.

[0348] Thus, administration of the composition to a subject may result in the following in a subject with type 2 diabetes and / or symptoms thereof: (a) reducing blood levels of one or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) may reduce the HOMA-IR score; and / or (c) It can increase blood adiponectin levels.

[0349] Thus, administration of the composition to a subject may result in the following in a subject with prediabetes and / or symptoms thereof: (a) reducing blood levels of one or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) may reduce the HOMA-IR score; and / or (c) It can increase blood adiponectin levels.

[0350] Those skilled in the art can easily diagnose type 2 diabetes and / or prediabetes using routine methods known in the art, for example, using reference values known in the art. For example, there are several test methods that can be used to determine whether a human subject has prediabetes and / or type 2 diabetes. Such test methods include, for example, A1C test, fasting plasma glucose test (FPG), urine glucose test, random blood glucose test, and oral glucose tolerance test (OGTT). Examples of reference values used in these methods are provided below. In some examples, several tests are used in combination to arrive at a diagnosis.

[0351] Type 2 diabetes is typically diagnosed using a glycosylated hemoglobin (A1C) test. The hemoglobin A1c (glycated hemoglobin, glycosylated hemoglobin, HbA1c, or A1c) test is used to assess an individual's level of glucose control. This test shows an average blood glucose level over the past 90 days, expressed as a percentage. For example, this blood test may show an individual's average blood glucose level over the past two to three months. Hemoglobin is a protein found only in red blood cells. Because red blood cells live an average of about three months, an A1c test reflects the red blood cells present in the bloodstream at the time of the test, and therefore, A1c is an average measure of blood glucose control. Hemoglobin's primary function is to transport oxygen from the lungs to cells throughout the body. Hemoglobin becomes glycosylated, or coated, by glucose in the bloodstream. Glucose present in the blood attaches to the hemoglobin protein, and increased glucose levels are reflected on the surface of the hemoglobin protein, resulting in elevated A1c levels. Typically, a blood sample, specifically a whole blood sample or a sample extracted from whole blood (e.g., serum and / or plasma), is used for the test (and diagnosis). Those skilled in the art will readily be able to identify the appropriate sample type depending on the parameter to be determined. Non-limiting examples of reference values that can be used in this test are as follows: less than 5.7% is normal, 5.7% to <6.5% (indicating impaired glucose regulation) is diagnostic of prediabetes, and 6.5% or greater in two separate tests indicates diabetes.

[0352] Type 2 diabetes and / or prediabetes can also be identified using a fasting blood glucose test. A fasting blood glucose test is a blood test that measures blood glucose levels after fasting. Fasting involves not eating or drinking anything (except water) for at least eight hours (e.g., until the test is completed). A blood sample is usually drawn after an overnight fast. Prolonged fasting triggers a hormone called glucagon, which is produced by the pancreas and causes the liver to release glucose (blood sugar) into the bloodstream. In the absence of diabetes, the body responds by producing insulin, which prevents hyperglycemia (high blood sugar). However, if the body cannot produce enough insulin or cannot respond appropriately to insulin, fasting blood glucose levels remain elevated. Typically, a blood sample, specifically a whole blood sample or a sample extracted from whole blood (e.g., serum and / or plasma), is used for testing (and diagnosis). Those skilled in the art can readily identify the appropriate sample type depending on the parameters to be determined. Non-limiting examples of how results may be interpreted are as follows: less than 100 mg / dL (5.6 mmol / L) is normal, 100 to <126 mg / dL (5.6 to 6.9 mmol / L) (indicating impaired fasting glucose / fasting hyperglycemia) is diagnostic of prediabetes, and 126 mg / dL (7 mmol / L) or greater on two separate tests is diagnostic of diabetes.

[0353] Type 2 diabetes and / or prediabetes can also be identified using an oral glucose tolerance test. A glucose tolerance test (GTT), also known as an oral glucose tolerance test (OGTT), is a method that can aid in diagnosing cases of diabetes mellitus or insulin resistance. A glucose tolerance test is used to determine an individual's ability to handle a glucose load, determining how quickly glucose is eliminated from the blood. This test can indicate whether an individual can metabolize a standardized amount of glucose. This test is not as widely used as other tests, except during pregnancy. Prior to the test, individuals are instructed not to eat or drink certain fluids for up to 8–12 hours before the test. They may also be instructed not to take certain medications prior to the test, but only if these medications may affect the test results. The test itself involves drawing blood to measure pre-test blood glucose levels. The next step involves consuming a very sweet-tasting glucose drink. Further blood samples are then drawn at regular intervals (30 or 60 minutes) or a single test is performed two hours later. Typically, a whole blood sample is used for testing (and diagnosis). Non-limiting examples of how the results can be interpreted are as follows: a fasting value (before the test) of less than 6 mmol / L and a two-hour value of less than 7.8 mmol / L is normal, a fasting value (before the test) of 6.0-7.0 mmol / L and a two-hour value of 7.8-11.1 mmol / L is diagnostic of pre-diabetes (indicating impaired glucose tolerance), and a fasting value (before the test) of greater than 7.0 mmol / L and a two-hour value of greater than 11.1 mmol / L is suggestive of diabetes.

[0354] Regarding normoglycemic (euglycemia) reference values, in some instances, the following cutoff values can be used when measurements are performed using the above-mentioned tests: A1C <5.7%, FPG <100 mg / dL, and OGTT 2-hour <7.8 mmol / L. Furthermore, normoglycemia (euglycemia) can be indicated by an interstitial glucose level of 70-180 mg / dL (where >180 mg / dL indicates hyperglycemia and <70 mg / dL indicates hypoglycemia).

[0355] As will be apparent to those skilled in the art, as used herein, "interstitial glucose" refers to glucose in interstitial fluid. Glucose is typically transported from the capillary endothelium into the interstitial fluid by simple diffusion down a concentration gradient, without the need for an active transporter. Blood flow to the site determines the amount of glucose delivered. Generally, interstitial glucose values (e.g., levels) are determined by the rate of glucose diffusion from plasma to the interstitial fluid and the rate of glucose absorption by cells in the subcutaneous tissue. Interstitial glucose levels can be measured using conventional methods known in the art. For example, interstitial glucose levels can be measured using a continuous glucose monitoring (CGM) device (such as that used in the Examples section below). The glucose sensors of these CGM devices are constructed as needle sensors that are inserted into subcutaneous adipose tissue to access interstitial fluid (ISF).

[0356] In some examples, the composition is for the treatment and / or prevention of type 2 diabetes and / or symptoms thereof.

[0357] Thus, in some examples, provided is a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for use in treating and / or preventing type 2 diabetes and / or symptoms thereof in a subject.

[0358] Also provided, in some examples, are methods for treating and / or preventing type 2 diabetes and / or symptoms thereof in a subject, comprising administering to the subject a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

[0359] As discussed elsewhere herein, the inventors have surprisingly shown that AB001, compared to placebo, has beneficial effects on glycemic control (measured by euglycemic hours, assessed by continuous glucose monitoring, and using biomarkers of glycemic control, i.e., HbA1c and fasting glucose) and on biomarkers of inflammation (e.g., chronic systemic inflammation, assessed using adiponectin or hsCRP as biomarkers), insulin resistance, and beta-cell dysfunction (assessed using blood insulin and intact proinsulin as biomarkers, and using HOMA-IR score) in patients with type 2 diabetes. The inventors have therefore advantageously demonstrated that the composition described herein (AB001) can be used to stabilize glucose levels and / or improve glycemic control in subjects with type 2 diabetes, prediabetes, and / or symptoms thereof. Advantageously, the compositions described herein may also be used to reduce insulin resistance, beta cell dysfunction, and / or inflammation (e.g., chronic systemic inflammation and / or intestinal inflammation) in subjects with type 2 diabetes, prediabetes, and / or symptoms thereof. In some examples, the inflammation may be intestinal and / or pancreatic inflammation. In addition, the inventors surprisingly observed that changes in lipid profiles (triglyceride levels, total cholesterol, and HDL cholesterol) and other metabolic syndrome parameters were more favorable with AB001 than with placebo in patients with type 2 diabetes. Advantageously, the compositions described herein may therefore be used for the treatment and / or prevention of symptoms of metabolic syndrome in subjects with type 2 diabetes and / or prediabetes. For further information, see Pfutzner et al, Fasting Intact Proinsulin Isa Highly Specific Predictor of Insulin Resistance in Type 2 Diabetes, DiabetesCare 27:682-687, 2004, which is incorporated herein by reference.

[0360] Symptoms of metabolic syndrome may be observed in patients with type 2 diabetes and / or prediabetes. As used herein, "metabolic syndrome" refers to a combination of insulin resistance, dyslipidemia, and hypertension. Symptoms of metabolic syndrome include, but are not limited to, hypertension, hypertriglyceridemia, hyperglycemia, hyperuricemia, and other metabolic disorders. The present invention may be used to improve symptoms of metabolic syndrome. As described elsewhere herein, in patients with type 2 diabetes, changes in lipid profiles (triglyceride levels, total cholesterol, and HDL cholesterol) and other metabolic syndrome parameters were more favorable with AB001 than with placebo. Advantageously, the compositions described herein may therefore be used for the treatment and / or prevention of symptoms of metabolic syndrome in subjects with type 2 diabetes and / or prediabetes.

[0361] Thus, in some examples, administering a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) comprises: (a) capable of reducing one or more levels of HbA1c or glucose; (b) may reduce fasting glucose levels; (c) may reduce insulin levels; (d) may reduce the HOMA-IR score for insulin resistance; (e) capable of lowering intact proinsulin; (f) may reduce blood hsCRP; and / or (g) It can increase blood adiponectin levels.

[0362] It has been suggested that one result of AB001 supplementation is a reduction in glucose absorption through the intestinal tract. Thus, in some examples, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces glucose absorption into the subject's blood. Those skilled in the art can determine the reduction in glucose absorption into the blood in the presence of the compositions provided herein (compared to when the composition is not used) using methods known in the art (e.g., by measuring blood glucose in the presence or absence of the compositions described herein using routine methods known in the art). A reduction in blood and interstitial fluid glucose by the composition would be considered by those skilled in the art to be an indicator of a significant improvement in macrovascular risk and overall health.

[0363] AB001 does not have an active hypoglycemic effect and does not affect normal glucose levels in healthy individuals or patients with diabetes. This has been observed in various clinical applications and in individual patient cases. In clinical trials, AB001 has not been associated with an increased risk of hypoglycemia, and the suggested mode of action is not consistent with these findings.

[0364] Thus, in some examples, administering a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) comprises: (a) reducing blood levels of one or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) reduce the HOMA-IR score, and / or (c) Increases blood adiponectin levels.

[0365] In some examples, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) comprises: (a) reducing blood levels of two or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) reduce the HOMA-IR score, and / or (c) Increases blood adiponectin levels.

[0366] In some examples, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) comprises: (a) reducing blood levels of three or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) reduce the HOMA-IR score, and / or (c) Increases blood adiponectin levels.

[0367] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood levels of one or more (e.g., two or more, or three or more) of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP. In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces HOMA-IR score.

[0368] Typically, the HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP levels in the subject's blood, and the HOMA-IR score are determined from a blood sample taken from the subject, particularly a whole blood sample or a sample extracted from whole blood (e.g., serum and / or plasma).Those skilled in the art can easily identify the type of sample that is suitable depending on the parameters to be determined.

[0369] The terms "reduce," "reduced," "lowering," "reduced," "decreased," or "downregulated," "lower" are all used generally herein to mean a decrease (e.g., by an amount (e.g., a statistically significant amount)). For the avoidance of doubt, in the context of the present invention, "reduced," "lowering," "decreased," "reduced," or "decreasing" means a decrease compared to a reference level / control.

[0370] The reference level / control in the context of the present invention may be the level before administration of the compositions described herein.

[0371] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood HbA1c levels. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood HbA1c levels compared to the blood HbA1c level before administration of the composition. HbA1c levels can be determined using routine methods known in the art, for example, an HbA1C test, as described in the Examples section below. HbA1c and HbA1C tests are discussed in more detail elsewhere herein. HbA1c levels can be used to identify impaired glucose regulation in a subject and, therefore, can be used as a biomarker for glycemic control, as described in the Examples section below.

[0372] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood glucose (e.g., fasting glucose) levels. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood glucose levels (e.g., after fasting) compared to blood glucose levels before administration of the composition (e.g., blood glucose levels after fasting before administration of the composition). Blood glucose can be determined using routine methods known in the art (as described in the Examples section below); for example, fasting blood glucose can be determined using an FPG test, which is discussed in more detail elsewhere herein. Fasting glucose levels can be used as a biomarker for glycemic control, as discussed in the Examples section below.

[0373] As used herein, "fasting blood glucose" refers to blood glucose levels after fasting. Typically, in the context of the present invention, fasting lasts 8-12 hours, e.g., 8-10 hours, during which no food or drink is consumed by the subject (although small amounts of water may be consumed in some cases). Thus, as used herein, "fasting blood glucose" typically refers to blood glucose levels after at least 8 hours of fasting, e.g., after 8-12 hours of fasting.

[0374] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood insulin levels. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood insulin levels compared to blood insulin levels before administration of the composition. Insulin is a peptide hormone secreted by beta cells in the islets of Langerhans in the pancreas and regulates blood glucose levels. Insulin acts by directly binding to its receptor on the cell membrane. These receptors are present on all cells, although their density varies depending on the cell type, with the highest density on hepatocytes and adipocytes. The insulin receptor is a heterotetrameric glycoprotein consisting of two subunits, an alpha subunit and a beta subunit. The extracellular alpha subunit contains the insulin-binding site. The transmembrane beta subunit has tyrosine kinase activity. When insulin binds to the alpha subunit, it activates the tyrosine kinase activity of the beta subunit, resulting in the translocation of a glucose transporter from the cytoplasm to the cell surface. These glucose transporters allow glucose to enter cells from the blood, thus lowering blood glucose levels.

[0375] Insulin has the following effects on cells: Hepatocytes: promotes glycogen production and inhibits gluconeogenesis, Adipocytes: promotes adipogenesis and inhibits lipolysis, Muscle cells: promotes glycogen production and protein synthesis and inhibits protein catabolism Pancreatic beta cells: inhibits glucagon release, · Brain cells: involved in appetite regulation.

[0376] Conditions such as diabetes tend to increase blood insulin levels. Those skilled in the art can easily determine blood insulin levels using conventional methods known in the art (as described in the Examples section below). As those skilled in the art will appreciate, blood insulin levels can be used as a biomarker for insulin resistance, as discussed in the Examples section below.

[0377] In some examples, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces the HOMA-IR score. In other words, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces the HOMA-IR score compared to the HOMA-IR score before administering the composition. "HOMA-IR" refers to insulin resistance assessed by homeostatic model assessment. This homeostatic model assessment insulin resistance (HOMA-IR), developed by Matthews et al., is widely used in research to evaluate insulin resistance. Compared to the "gold" standard for quantifying insulin resistance, quantification using HOMA-IR is more convenient. HOMA-IR is calculated by multiplying fasting plasma insulin (FPI) by fasting plasma glucose (FPG) and then dividing by the constant 22.5, i.e., HOMA-IR = (FPI x FPG) / 22.5. A low HOMA-IR value indicates high insulin sensitivity, while a high HOMA-IR value indicates low insulin sensitivity (insulin resistance).Those skilled in the art can easily determine fasting plasma insulin (FPI) level and fasting plasma glucose (FPG) level using conventional methods known in the art.Therefore, those skilled in the art can easily determine an individual's HOMA-IR score using conventional methods known in the art.

[0378] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces the level of intact proinsulin in the blood. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces the level of intact proinsulin in the blood compared to the level of intact proinsulin in the blood before administration of the composition. Proinsulin is produced in pancreatic beta cells and is normally further processed into insulin and C-peptide. Proinsulin is found in low concentrations in the blood (e.g., plasma) of healthy subjects. Increased insulin demand caused by insulin resistance in late-stage type 2 diabetes mellitus can result in increased proinsulin expression in the blood. Intact proinsulin is rapidly degraded. In clinical practice, first-morning fasting intact proinsulin can be used as a highly specific indicator of clinically relevant insulin resistance to select therapies for insulin resistance and to monitor the effectiveness of treatment of beta cell dysfunction. Those skilled in the art can readily determine the level of intact proinsulin in the blood using routine methods known in the art (as described in the Examples section below). As those skilled in the art will appreciate, the level of intact proinsulin can be used as a biomarker of beta cell dysfunction and / or insulin resistance.

[0379] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood hsCRP levels. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood hsCRP levels compared to blood hsCRP levels before administration of the composition. C-reactive protein (CRP) is a blood protein that increases in the presence of inflammation. CRP has long been used as an indicator of disease-related infection and inflammation. hs-CRP tests accurately measure low levels of CRP to identify low but persistent levels of inflammation. Therefore, as will be appreciated by those skilled in the art, hsCRP levels can be used as a biomarker of inflammation, particularly chronic systemic inflammation. Those skilled in the art can readily determine blood hs-CRP levels using routine methods known in the art (as described in the Examples section below).

[0380] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) increases blood adiponectin levels. In other words, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) increases blood adiponectin levels compared to blood adiponectin levels before administration of the composition. Adiponectin is the most abundant peptide hormone secreted by adipocytes. Adiponectin is a hormone released by adipose (fat) tissue and is useful for insulin sensitivity and inflammation. Low adiponectin levels are associated with several conditions, including type 2 diabetes and atherosclerosis. Adiponectin can be used as a biomarker of inflammation, particularly chronic systemic inflammation. Those skilled in the art can easily determine blood adiponectin levels using routine methods known in the art (as described in the Examples section below).

[0381] The terms "increased", "increase", or "upregulated", "higher" are all used generally herein to mean an increase (e.g., by an amount (e.g., a statistically significant amount). For the avoidance of doubt, in the context of the present invention, the term "increased" or "increase" means an increase compared to a reference level / control.

[0382] In the context of the present invention, the reference level / control may be the level prior to administration of the compositions described herein.

[0383] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) controls blood glucose (e.g., whole blood glucose, plasma glucose, and / or serum glucose) and / or interstitial glucose in the subject.

[0384] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) controls blood glucose (e.g., whole blood glucose, plasma glucose, and / or serum glucose) in the subject.

[0385] In some examples, administration of the composition to a subject (eg, a subject with type 2 diabetes, pre-diabetes, and / or symptoms thereof) controls interstitial glucose in the subject.

[0386] Thus, in certain examples, administration of a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) stabilizes blood glucose levels and / or interstitial glucose levels in the subject. In certain examples, administration of a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) stabilizes blood glucose levels in the subject. In other examples, administration of a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) stabilizes interstitial glucose levels in the subject.

[0387] As discussed elsewhere herein, it is suggested that AB001 has four glucose-controlling effects. For example, it is suggested that the Bacilli strains of AB001 and their secreted enzymes act as additional glucose digestion factors (in other words, the Bacilli strains of AB001 and their secreted enzymes act in addition to the subject's microbiome (e.g., gut microbiome) present prior to ingestion of AB001), further reducing glucose reabsorption into the blood. Thus, in some instances, administration of a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) degrades glucose. In some instances, administration of a composition to a subject degrades glucose within the subject's gut. In some instances, administration of a composition to a subject degrades glucose within the subject's intestine (particularly within the small intestine). More specifically, administration of a composition used in accordance with the present invention to a subject degrades glucose within the subject's duodenum. Glucose degradation and glucose-degrading compositions used in accordance with the present invention are described elsewhere herein. The descriptions of "glucolytic" and "glucolytic composition" provided elsewhere herein apply equally here.

[0388] As discussed elsewhere herein, glucose may be metabolized (i.e., glucose may be broken down by metabolism). Thus, in some examples, administration of a composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) metabolizes glucose. In some examples, administration of a composition to a subject metabolizes glucose in the subject's digestive tract. "Metabolism" is defined elsewhere herein and applies equally here. In some examples, administration of a composition used in accordance with the present invention to a subject metabolizes glucose in the subject's intestine. In one example, administration of a composition used in accordance with the present invention to a subject metabolizes glucose in the subject's small intestine. More specifically, administration of a composition used in accordance with the present invention to a subject metabolizes glucose in the subject's duodenum.

[0389] It has been suggested that one result of AB001 supplementation is a reduction in glucose absorption through the intestinal tract. Thus, in some examples, administering the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces the subject's glucose absorption into the blood. Those skilled in the art can use methods known in the art (e.g., by measuring blood glucose in the presence or absence of the compositions described herein using conventional methods known in the art) to determine the reduction in glucose absorption into the blood in the presence of the compositions provided herein (compared to when the composition is not used).

[0390] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces glucose absorption into the interstitial fluid of the subject. One of ordinary skill in the art would be able to determine the reduction in glucose absorption into interstitial fluid in the presence of a composition provided herein (compared to when the composition is not used) using methods known in the art (e.g., by measuring interstitial glucose in the presence or absence of a composition described herein using routine methods known in the art).

[0391] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood glucose levels and / or interstitial glucose levels in the subject. One of ordinary skill in the art would be able to determine the reduction in blood glucose levels and / or interstitial glucose levels in the presence of a composition provided herein (compared to when the composition is not used) using methods known in the art.

[0392] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces blood glucose levels in the subject. One of ordinary skill in the art would be able to determine the reduction in blood glucose levels in the presence of a composition provided herein (compared to when the composition is not used) using methods known in the art.

[0393] In some examples, administration of the composition to a subject (e.g., a subject with type 2 diabetes, prediabetes, and / or symptoms thereof) reduces interstitial glucose levels in the subject. One of ordinary skill in the art would be able to determine the reduction in interstitial glucose levels in the presence of a composition provided herein (compared to when the composition is not used) using methods known in the art.

[0394] The compositions provided herein are effective when administered to a subject for an appropriate length of time and at an appropriate frequency, which can be readily determined by one skilled in the art or a corresponding physician.

[0395] In some examples, a composition (e.g., an effective amount of a composition described herein) is ingested. In some examples, a composition (e.g., an effective amount of a composition described herein) is ingested daily for an appropriate length of time. In other words, a composition used in accordance with the present invention may be for daily administration.

[0396] In some examples, the composition (e.g., an effective amount of a composition described herein) is taken for at least one day, hi some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least two days, at least three days, at least four days, at least five days, at least six days, or at least seven days.

[0397] In some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least 1 week. In some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least 2 weeks. In some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least 4 weeks. In some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least 6 weeks. In some examples, the composition (e.g., an effective amount of a composition described herein) is taken daily for at least 6 months.

[0398] For example, one dose (e.g., two capsules or two tablets of a composition described herein) can be taken daily for at least two days, at least three days, at least four days, at least five days, at least six days, or at least seven days. For example, one dose (e.g., two capsules or two tablets of a composition described herein) can be taken daily for two, three, four, five, six, or seven days.

[0399] For example, one dose (e.g., two capsules or two tablets of a composition described herein) can be taken daily for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks. For example, one dose (e.g., two capsules or two tablets of a composition described herein) can be taken daily for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks.

[0400] "Ingestion," as used herein, refers to the uptake (e.g., consumption) of a substance by an organism (e.g., a subject, preferably a human). In the context of the present invention, ingestion is preferably oral, and the subject is a human.

[0401] As will be apparent to one of skill in the art, the compositions used in accordance with the invention described herein may be used in combination with one or more other therapies (e.g., drugs) used in the prevention and / or treatment of type 2 diabetes, prediabetes, and / or symptoms thereof. Thus, the compositions used in accordance with the invention described herein may be used to complement one or more other therapies (e.g., drugs) used in the prevention and / or treatment of type 2 diabetes, prediabetes, and / or symptoms thereof.

[0402] In one example, a composition used in accordance with the invention described herein can be administered to (e.g., ingested by) a subject undergoing treatment with one or more therapies (e.g., drugs) used to prevent and / or treat type 2 diabetes, prediabetes, and / or symptoms thereof. Therapies (e.g., drugs) used to prevent and / or treat type 2 diabetes, prediabetes, and / or symptoms thereof are well known in the art. Non-limiting examples of therapies (e.g., drugs) used to prevent and / or treat type 2 diabetes, prediabetes, and / or symptoms thereof are described in the Examples section below.

[0403] Compositions used in accordance with the present invention (e.g., compositions used to treat and / or prevent type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject, and / or compositions used in methods of treating and / or preventing type 2 diabetes, pre-diabetes, and / or symptoms thereof in a subject) may be taken orally in the form of an aqueous solution, tablet, capsule, or granule, etc.

[0404] The compositions described herein are for administration to a subject (preferably a human) in an effective amount (effective dose). An "effective amount" is an amount that, alone or in combination with further doses, produces the desired (therapeutic or non-therapeutic) response. The effective amount used will depend, for example, on the therapeutic (or non-therapeutic) purpose, the route of administration, and the condition of the subject. For example, the dosage of the compositions of the present invention appropriate for a given subject can be determined by a physician (or the person administering the composition) taking into account various factors known to alter the action of the compositions of the present invention, such as body weight, sex, diet, time and route of administration, other medications, and other relevant clinical factors. The dosage and schedule can vary depending on the specific condition, disorder, or overall symptom state of the subject. The effective dosage can be determined by in vitro or in vivo methods.

[0405] The compositions described herein are advantageously presented in dosage units. For example, the compositions may be presented in the form of capsules or tablets. Other suitable dosage units are described elsewhere herein.

[0406] An "effective amount" can include administration of one or more dosage units. For example, an effective amount can be achieved by administration of one or two capsules or tablets. When an effective amount includes multiple dosage units, the dosage units can be administered at once or taken at intervals throughout the day.

[0407] Alternate appropriate effective amounts and dosage forms will be readily ascertainable by those of ordinary skill in the art using routine experimentation based on the examples below.

[0408] As used herein, the terms "treat" and "treatment" refer to administering a composition to a subject (e.g., a symptomatic subject suffering from an adverse condition, disorder, illness, or disease (e.g., type 2 diabetes, prediabetes, and / or symptoms thereof)) to thereby result in a decrease in the severity and / or frequency of the symptoms, eliminate the symptoms and / or their underlying causes, and / or promote amelioration or repair of damage, and / or prevent the adverse condition, disorder, illness, or disease in a symptomatic subject susceptible to, or suspected of having or at risk of developing, a particular adverse condition, disorder, illness, or disease.

[0409] By way of example, as described elsewhere herein, treatment with the compositions described herein advantageously includes: (a) reducing blood levels of one or more of HbA1c, glucose (e.g., fasting glucose), insulin, intact proinsulin, or hsCRP; (b) may reduce the HOMA-IR score; and / or (c) It can increase blood adiponectin levels.

[0410] Treatment with the compositions described herein may also advantageously control blood glucose, reduce glucose absorption into the blood of a subject, and / or reduce blood glucose concentrations in a subject.

[0411] The term "prevention," as used herein, refers to the avoidance of the occurrence or recurrence of a symptom and / or its underlying cause, injury, adverse condition, disorder, illness, and / or disease. For example, "prevention" in the context of the present invention can be the avoidance of the occurrence of one or more of the symptoms associated with type 2 diabetes and / or prediabetes.

[0412] As discussed elsewhere herein, the subject can be any suitable subject, for example, the subject can be a human. Preferably, the subject is a human. The subject can be a human with type 2 diabetes, prediabetes, and / or symptoms thereof. A physician can readily diagnose type 2 diabetes, prediabetes, and / or symptoms thereof using routine methods known in the art, for example, using the tests and reference values discussed elsewhere herein. The subject can be a human who has been diagnosed with type 2 diabetes, prediabetes, and / or symptoms thereof, and who is about to or has taken glucose.

[0413] Unless otherwise defined herein, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. For example, Singleton and Sainsbury, Dictionary of Microbiology and Molecular Biology, 2nd Ed., John Wiley and Sons, NY (1994), and Hale and Marham, The Harper Collins Dictionary of Biology, Harper Perennial, NY (1991), provide those skilled in the art with a comprehensive dictionary of many of the terms used in this invention. In practicing the present invention, any methods and materials similar or equivalent to those described herein can be used; however, preferred methods and materials are described herein. Accordingly, the terms defined below are more fully described with reference to the specification as a whole. Also, as used herein, the singular terms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Unless otherwise indicated, nucleic acids are written left to right in 5' to 3' orientation, and amino acid sequences are written left to right in amino to carboxy orientation, respectively. It is understood that this invention is not limited to the particular methodology, protocols, and reagents described, as these may vary with the context in which they are used by those of skill in the art.

[0414] Aspects of the present invention are illustrated by the following non-limiting examples. [Example]

[0415] Development of the compositions described herein The inventors previously developed a food-grade fermented rice bran for evaluation purposes. A safety study was initiated using a case study design. Approximately 1,400 people took the product regularly over a long period of time. No side effects / adverse reactions were reported.

[0416] Initial indications of an effect on glucose were obtained from a case-study safety study of oral administration of fermented rice bran containing Bacilli strains (containing no L-cysteine and no dextrin). Patients with type 2 diabetes reported an extended period of euglycemia (self-testing) and, during routine health checkups every 3 to 6 months, increased HDL levels, decreased triglyceride levels, and decreased HbA1c levels. At the physician's discretion, some of these individuals discontinued one daily dose of a sensitizer, and some reduced or completely discontinued their statin dose. After 3 to 6 months, some patients with type 2 diabetes demonstrated HbA1c and other blood parameters typical of prediabetes. (Data not shown.) This safety study demonstrates that a composition containing fermented rice bran and Bacilli strains (containing no L-cysteine and no dextrin) is beneficial for patients with type 2 diabetes.

[0417] These findings led to the formulation of AB001, a fermented rice bran containing up to approximately 70% Bacilli strains, which was evaluated in a randomized, placebo-controlled clinical trial and demonstrated positive effects on euglycemia, LDL-HDL ratio, triglycerides, HbA1c, etc. The results of this clinical trial are shown in Example 1.

[0418] Prediabetes is a precursor to type 2 diabetes, and therefore, from a scientific and clinical perspective, the data discussed herein demonstrate that fermented rice bran / Bacilli strains may be a highly efficient and effective treatment for prediabetes (the composition may be used to avoid / prevent medical conditions that lead to type 2 diabetes).

[0419] The composition (AB001) described herein typically consists of naturally fermented rice bran, Bacillus subtilis and Bacillus coagulans, L-cysteine, and dextrin. The composition may also contain magnesium stearate, and calcium and potassium phosphates. This supplement is an acid-resistant HPMC capsule that dissolves upon reaching the duodenum. The culture is released and settles in the upper intestinal tract, where it remains for approximately one day before being excreted from the body through feces. The bacterial strain included in the composition was selected to favorably and efficiently metabolize glucose.

[0420] The bacterial composition of AB001 was investigated using Sanger DNA sequencing. The predominant bacterial strains found in the composition were identified as Bacillus subtilis spp. and Bacillus coagulans spp. Other genera / species identified were Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp. MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus.

[0421] Dextrins from three different suppliers were tested and found to work in the compositions of the present invention (data not shown). The preferred dextrin used herein was hydrolyzed corn dextrin (e.g., Vitargo). L-cysteine from three different suppliers was also tested and found to work (data not shown). The preferred L-cysteine used herein was of plant origin.

[0422] The composition used in Example 1 contained, per dose (2 capsules), fermented rice bran (552 mg), L-cysteine (150 mg), microcrystalline cellulose (40 mg), magnesium stearate (4.8 mg), dextrin (4 mg), silicon dioxide (4 mg), and at least 1×10 5The formulation contained 10 ...

[0423] Example 1 - Effect of a bacteria-based nutritional supplement (AB001) on glycemic control and metabolic syndrome biomarkers in patients with type 2 diabetes mellitus summary BACKGROUND:AB001 is a nutritional supplement based on a microbial consortium of commensal bacterial strains. The aim of this study was to investigate the potential beneficial effects of the supplement compared with placebo on glycemic control and biomarkers of inflammation, insulin resistance, and beta-cell dysfunction.

[0424] Methods: A total of 40 patients with type 2 diabetes (31 men, 9 women, age: 65.5 ± 8.0 years, BMI: 33.6 ± 5.5 kg / m) were included. 2 , HbA1c: 7.2±0.9%) were enrolled in this study. Patients were randomized to receive either AB001 or placebo once daily for six weeks. Specifically, participants in the treatment arm of this clinical trial took two capsules of the composition (e.g., one dose of the composition) per day. Observation parameters included euglycemic time assessed by continuous glucose monitoring, as well as biomarkers of glycemic control, beta cell function, insulin resistance, and chronic systemic inflammation.

[0425] Results: Time in range remained stable at weeks 0-2 vs. weeks 4-6 with AB001, but slightly worsened (-4%) with placebo. At endpoint, AB001, but not placebo, significantly improved biomarkers of glycemic control (HbA1c: -0.3% vs. 0.1%, fasting glucose: -14% vs. +8%, all p<0.05), insulin resistance (insulin: -17% vs. +14%, HOMA-IR: -26% vs. +21%, all p<0.05), beta-cell dysfunction (intact proinsulin: -40% vs. -2%, p<0.05), and chronic systemic inflammation (adiponectin: +8% vs. -8%, p<0.05, hsCRP: -31% vs. +27%, not significant). Additionally, changes observed in lipid profiles and other metabolic syndrome parameters in patients with TIID were more favorable with AB001 than with placebo. There were no differences between groups regarding the number and type of adverse events.

[0426] Conclusions: The results observed with AB001 indicate an overall improvement in cardiometabolic status. AB001 is a valuable supplement to any existing concomitant treatment in patients with type 2 diabetes.

[0427] Introduction As described elsewhere herein, de Faire Medical AB (Stockholm, Sweden, www.defairemedical.com) has developed AB001, a nutritional supplement based on fermented rice bran containing Bacillus subtilis and Bacillus coagulans, which contribute to the product's prebiotic and probiotic properties. The product's action is driven, in part, by the secretion of bioactive substances, such as enzymes, that break down complex organic molecules into nutrients, reactants, and energy. For example, bacterial strains, such as Bacillus subtilis and B. coagulans, contained in the product in combination with dextrin and L-cysteine favorably metabolize glucose and other more complex carbohydrate molecules. A more detailed mechanism of action is described elsewhere herein. AB001 does not have an active blood sugar-lowering effect and does not affect normal glucose levels in healthy individuals or patients with diabetes. The product complies with European regulatory frameworks as a nutritional supplement. In this double-blind study, patients with type 2 diabetes mellitus in the late stage of the disease received either AB001 or placebo as a nutritional supplement for six weeks in addition to standard of care intervention.

[0428] Patients and methods This prospective, double-blind, placebo-controlled, single-center, parallel-group study was conducted in accordance with international and local ethical and scientific standards. The protocol was approved by the competent ethical review committee (Landesarztekammer Rheinland-Pfalz, Mainz, Germany) and reported to the national competent authority (Bundesamt fur Verbraucherschutz und Lebenmittelsicherheit). The study was registered with the German Registry for Clinical Studies (DRKS number: DRKS00023745).

[0429] The primary objective was the effect of AB001 on glycemic control (time to normal glucose) compared with placebo and assessed by continuous glucose monitoring with the Freestyle Libre 2 device (Abbott Diagnostics, Wiesbaden-Delkenheim, Germany) after 2, 4, and 6 weeks of treatment. Secondary objectives were the effect of the supplement compared with placebo on HbA1c, lipids, uric acid, insulin sensitivity (assessed by the HOMA score

[16] ), beta-cell dysfunction (insulin and intact proinsulin [17,18]), chronic systemic inflammation, and cardiovascular risk (total adiponectin and hsCRP [19,20]), as well as the tolerability of the supplement (type and nature of (serious) adverse events).

[0430] To be eligible for the study, patients had to be adults and had a diagnosis of type 2 diabetes (HbA1c 6.0%-9.9% and treated with any type or combination of standard therapy). They could not have type 1 diabetes, any acute or chronic gastrointestinal disease, anemia, acute hyperthyroidism, a history of allergies to probiotic supplements, or any serious life-threatening illness.

[0431] Prior to enrollment, participants signed written informed consent. Blood samples were then collected for safety analysis and to identify potential exclusion criteria. At the baseline visit (V1), randomization into two study groups and blood samples were collected for efficacy parameter determination. The overall observation period was 6 weeks. A subcutaneous continuous glucose monitoring (CGM, FreeStyle Libre, Abbott) sensor was provided for the next 2 weeks. Three follow-up visits (V2, V3, and V4) were scheduled at 2-week intervals to replace the CGM sensor and collect blood samples for efficacy analysis. Each patient's participation in the study was terminated at the end of the fourth visit.

[0432] All safety and efficacy parameters were measured in a central clinical laboratory. Insulin, intact proinsulin, and total adiponectin were determined using immunoassays (TecoMedical, Sissach, CH). Lipids, hsCRP, glucose, and HbA1c were determined with standard laboratory analyzers (Cobas c513, Roche Diagnostics, Basel, CH). At each treatment visit, patients were asked if any (serious) adverse events had occurred.

[0433] Data were evaluated using standard exploratory and descriptive analytical methods to gain an understanding of the qualitative and quantitative nature of the collected data. For quantitative variables, arithmetic means, medians, standard deviations, and minimum and maximum values were determined. Appropriate parametric and nonparametric statistical tests were used to compare collected results. For normally distributed data sets, Student's t-tests were used to compare changes from baseline to endpoint, and between groups for changes from baseline. A p-value of <0.05 was considered statistically significant.

[0434] The composition used contained, per dose (2 capsules), fermented rice bran (552 mg), L-cysteine (150 mg), microcrystalline cellulose (40 mg), magnesium stearate (4.8 mg), dextrin (4 mg), silicon dioxide (4 mg), and at least 1×10 5 The formulation contained 10 ...

[0435] Two capsules of the composition (eg, one dose of the composition) were taken daily by participants in the treatment arm of the trial.

[0436] result A total of 46 patients were screened, of which 40 who met the inclusion / exclusion criteria were able to participate in this study (31 men, 9 women, mean age: 65.4 ± 8.0 years (range: 44 to 89 years), BMI: 33.3 ± 5.3 kg / m 2 (Range: 21.5 kg / m 2 ~42.8kg / m 2 ). HbA1c: 7.2 ± 0.9% (55 ± 10 mmol / mol, range: 6.0%-9.4% / 42-79 mmol / mol). All enrolled patients completed the study according to protocol. After randomization, there were slightly more women in the placebo group. The majority of patients were receiving concomitant antidiabetic medications, indicating advanced disease. Baseline patient characteristics in both treatment groups, as well as the nature and distribution of diabetes treatment, are provided in Table 1.

[0437] The primary measured variable was euglycemic time as determined by FreeStyle Libre analysis software between weeks 0-2 and 4-6. Note that data were not available before patients began taking the supplement in this study; therefore, there was no true baseline for comparison to understand the direct impact of AB001 supplementation on euglycemic time. A comparison of the initial and final sensor analyses in this trial is provided in Figure 1.

[0438] It can be seen that patients taking AB001 had stable glycemic control, while a slight, but non-significant, deterioration was observed in the placebo group.

[0439] The differences observed for the majority of clinical parameters (weight, blood pressure, waist circumference) were numerically favorable to AB001 but were not significant. Only the observed reduction in waist circumference was significantly greater with AB001 (121±11 cm to 116±11 cm, p<0.005) compared to placebo (116±11 cm vs. 116±10 cm, not significant).

[0440] A panel of biochemical parameters was measured at baseline and endpoint to explore the effect of AB001 on glycemic control and biomarkers of underlying diabetic exacerbation (insulin resistance, beta-cell dysfunction, and chronic systemic inflammation). The results are provided in Table 2, and the percent change from baseline for both groups is provided in Figure 2.

[0441] Both hemoglobin A1c and fasting glucose improved significantly with AB001, whereas placebo showed no change (HbA1c) or worsening (fasting glucose).

[0442] Intact proinsulin, insulin, and HOMA-IR scores were determined as indicators of insulin resistance and beta-cell dysfunction [16-18]. These all significantly improved with AB001, but worsened with placebo. All changes from baseline with AB001, as well as between-group differences, reached statistical significance. These findings demonstrate the significant positive impact of AB001 on insulin resistance in this trial.

[0443] The biomarkers of chronic systemic inflammation determined in this study were total adiponectin and hsCRP [19,20]. Adiponectin significantly increased with AB001 and decreased with placebo. These observations were also highly significant for the between-group changes (p<0.001). AB001 resulted in a >30% decrease in hsCRP levels, whereas placebo increased it by >20%. However, due to the large variability observed, the between-group changes did not reach statistical significance.

[0444] A similar picture was observed for other laboratory biomarkers of metabolic syndrome (triglycerides, cholesterol, and uric acid) (see Table 2). There was a statistically significant improvement in triglyceride levels with AB001, but not with placebo. There was a strong trend in favor of AB001 for total cholesterol and HDL cholesterol levels. The deterioration observed for LDL cholesterol and uric acid was less pronounced with AB001 than with placebo. These findings are consistent with the generally positive laboratory picture observed with AB001 compared with placebo.

[0445] The supplement was well tolerated, and no serious adverse events were reported in this study. A total of 33 adverse events were reported, of which 18 occurred in the placebo group and 15 in the AB001 group. Symptoms related to gastrointestinal problems were observed in six patients in the placebo group and only one in the AB001 group. Hypoglycemia was reported six times by two patients in the AB001 group and four times by four patients in the placebo group. Based on the adverse events reported, it can be concluded that the nutritional supplement did not cause any particular side effects in this study. In addition, no other safety biochemistry panel values measured before and after the study showed any clinically relevant changes during the study.

[0446] Consideration Diabetes mellitus (DM) is a chronic systemic disease with complications affecting most body organs. It is widespread, affecting approximately 9% of the world's population

[21] . Alterations in pancreatic β-cell function and insulin resistance result in relative insulin deficiency and impaired cellular response to insulin. A key symptom of the disease is hyperglycemia, which can lead to several serious secondary complications [22,23]. Antidiabetic medications are commonly used according to national and international treatment guidelines [24,25]. The most commonly used drug, metformin, may have limited efficacy and may have adverse effects on the composition of the intestinal bacterial microfauna. Recently, alterations in the gastrointestinal microbiome have been found in diabetic patients, including an increased ratio of bacteroids to clostridium species in the gastrointestinal tract and an increased number of various opportunistic pathogens [26-29]. Circulating gram-positive GI bacteria have been detected in blood samples from diabetic patients

[28] . Alterations in the gut microbiome can lead to metabolic endotoxemia through lipopolysaccharide release, thus stimulating inflammation and insulin resistance

[30] . Epithelial enteroendocrine L cells play a role in inducing inflammation, and their number has been positively or negatively correlated with the abundance of 25 bacterial taxa in the gut [31, 32].

[0447] Given these findings, the complementary effects of probiotics on diabetes and diabetes-related complications have become the focus of modern research. Animal studies have shown positive results, including reductions in blood glucose levels, hemoglobin A1c (HbA1c), and insulin resistance [33,34]. Early human trials have also investigated the clinical effects of probiotics on glycemic control [14,35,36]. In these placebo-controlled trials, probiotic fermented milk, probiotic yogurt, bread, and kefir, primarily containing different Lactobacillales strains, and brewer's yeast were administered for 6–12 weeks. A meta-analysis of these trials reported significant improvements in HbA1c, fasting glucose, and HOMA-IR scores, but nonsignificant improvements were seen in insulin and fasting proinsulin levels

[37] .

[0448] The development of AB001 differs from these interventions in that a specific bacterial strain was selected that preferably metabolizes glucose and other carbohydrates and should therefore have a more potent effect on glucose levels and insulin resistance. Within the limitations of the present study protocol, the hypothesis of this more pronounced effect is substantiated, particularly by employing a panel of laboratory biomarkers that describe the diabetic phenotype by indicating the severity of the underlying diabetic exacerbation.

[38]

[0449] One consequence of AB001 supplementation appears to be a reduction in glucose absorption through the intestinal tract. Similar effects and corresponding biochemical findings were observed when drugs belonging to the SGLT-1 inhibitor class (α-glucosidase inhibitors, e.g., acarbose or voglibose) were used in routine diabetes treatment. The difference between these two approaches is that SGLT-1 inhibitors actively retain glucose molecules in the intestinal tract, where they are ultimately metabolized by resident bacteria in the large intestine, with methane gas as the end product. Therefore, patients treated with SGLT-1 inhibitors consistently complained of significant problems with flatulence and abdominal bloating [39,40]. Many patients in both treatment groups were already using insulin on different treatment regimens, indicating some degree of disease progression. The positive effects of AB001 on biomarkers of underlying diabetic exacerbation must have been quite pronounced, with consistently positive and significant results observed in almost all of the measures examined.

[0450] conclusion The results observed with AB001 indicated an overall improvement in cardiometabolic status, whereas the opposite was observed for placebo across all parameters. Considering the intensive background diabetes treatment the participants were receiving, these results are a strong indication that AB001 may be a valuable supplement to any existing concomitant treatment in patients with type 2 diabetes.

[0451] References 1.Gentile CL, Weir TL. The gut microbiota at the intersection of diet and human health. Science. 2018; 362:776-780. 2.Lynch SV, Pedersen O. The HumanIntestinal Microbiome in Health and Disease. N Engl J Med. 2016; 375:2369-2379. 3.Postler, T.S.; Ghosh, S. Understandingthe Holobiont: How Microbial Metabolites Affect Human Health and Shape theImmune System. Cell Metab.2017, 26:110-130. 4.Bibbo, S.; Ianiro, G.; Giorgio, V.; Scaldaferri,F.; Masucci, L.; Gasbarrini, A.; Cammarota, G. The role of diet on gutmicrobiota composition.Eur. Rev. Med. Pharmacol. Sci.2016,20, 4742-4749. 5.Sender, R.; Fuchs, S.; Milo, R. RevisedEstimates for the Number of Human and Bacteria Cells in the Body. PLoSBiol.2016,14, e1002533. 6.Kahrstrom, C.T.; Pariente, N.; Weiss, U.Intestinal microbiota in health and disease. Nature 2016, 535:47 7.Macfarlane, S.; Macfarlane, G.T.Regulation of short-chain fatty acid production. Proc. Nutr. Soc.2003, 62:67-72 8.Rowland, I.; Gibson, G.R.; Heinken, A.;Scott, K.P.; Swann, J.; Thiele, I.; Tuohy, K.M. Gut microbiota functions:Metabolism of nutrients and other food components. Eur. J. Nutr.2018, 57:1-24 9.Patterson E, Ryan PM, Cryan JF, Dinan TG,Ross RP, Fitzgerald GF, Stanton C.Gut microbiota, obesity and diabetes.Postgrad Med J. 2016;92:286-300. 10.Ortega MA, Fraile-Martinez O, Naya I,Garcia-Honduvilla N, Alvarez-Mon M, Bujan J, Asunsolo A, de la Torre B. Type 2Diabetes Mellitus Associated with Obesity (Diabesity). The Central Role of GutMicrobiota and Its Translational Applications. Nutrients. 2020; 12:2749 11.Castro-Barquero S, Lamuela-Raventos RM,Domenech M, Estruch R. Relationship between Mediterranean Dietary PolyphenolIntake and Obesity.Nutrients 2018;10:1523. 12.Roberfroid M, Gibson GR, Hoyles L,McCartney AL, Rastall R, Rowland I, Wolvers D, Watzl B, Szajewska H, Stahl B,et al. Prebiotic effects: Metabolic and health benefits. Br. J.Nutr.2010;104:S1-S63 13.Abraham BP, Quigley EMM. Probiotics inInflammatory Bowel Disease. Gastroenterol Clin North Am. 2017; 46:769-782. 14.Asemi Z, Samimi M, Tabassi Z, et al.Effect of daily consumption of probiotic yoghurt on insulin resistance inpregnant women: a randomized controlled trial. Eur J Clin Nutr. 2013;67:71-74 15.Wilkins T, Sequoia J.Probiotics forGastrointestinal Conditions: A Summary of the Evidence. Am Fam Physician. 2017;96:170-178. 16.Matthews DR, Hosker JP, Rudenski AS,Naylor BA, Treacher DF, Turner RC. Homeostasis model assessment: insulinresistance and β-cell function from fasting plasma glucose and insulinconcentrations in man. Diabetologia 1985; 28:412-419 17.Pfutzner A., Kunt T., Mondok A., PahlerS., Konrad T., Luebben G., Forst T.: Fasting Intact Proinsulin is a HighlySpecific Predictor of Insulin Resistance in Type 2 Diabetes. Diabetes Care2004; 27:682-687 18.Pfutzner A., Kann P., Pfutzner A.H.,Kunt T., Larbig M., Weber M.M., Forst T.: Intact and total proinsulin: newaspects for diagnosis and treatment of type 2 diabetes. Clin. Lab. 2004;50:567-573 19.Schondorf T., Maiworm A., Emisson N.,Forst T., Pfutzner A.: Biological Background and Role of Adiponectin as Markerfor Insulin Resistance and Cardiovascular Risk. Clin Lab. 2005; 51:489-494 20.Pfutzner A., Forst T.: HsCRP ascardiovascular risk marker in patients with diabetes mellitus. Diab. Technol.Ther. 2006; 8:28-36 21.International Diabetes Federation. IDFDiabetes Atlas 2019 https: / / www.diabetesatlas.org 22.Cree-Green M, Triolo TM, Nadeau KJ.Etiology of insulin resistance in youth withtype 2 diabetes. Curr Diab Rep.2013;13:81-88. 23.Mizokami-Stout K, Cree-Green M, NadeauKJ. Insulin resistance in type 2 diabetic youth. Curr Opin Endocrinol DiabetesObes. 2012;19:255-262. 24.American Diabetes Association.Pharmacologic Approaches to Glycemic Treatment: standards of Medical Care inDiabetes-2021. Diabetes Care. 2021; 44(Suppl.1):S111-S124 25.Davies MJ, D'Alessio DA, Fradkin J,Kernan WN, Mathieu C, Mingrone G, Rossing P, Tsapas A, Wexler DJ, Buse JB.Management of Hyperglycemia in Type 2 Diabetes, 2018. A Consensus Report by theAmerican Diabetes Association (ADA) and the European Association for the Studyof Diabetes (EASD). Diabetes Care. 2018; 41:2669-2701 26.Qin J, Li Y, Cai Z, et al. Ametagenome-wide association study of gut microbiota in type 2 diabetes. Nature.2012;490:55-60. 27.Larsen N, Vogensen FK, van den Berg FW,et al. Gut microbiota in human adults with type 2 diabetes differs fromnon-diabetic adults. PLoS One. 2010;5:e9085. doi: 10.1371 / journal.pone.0009085. 28.Sato J, Kanazawa A, Ikeda F, et al. Gutdysbiosis and detection of “live gut bacteria” inblood of Japanese patients with type 2 diabetes. Diabetes Care.2014;37:2343-2350. 29.Soyucen E, Gulcan A, Aktuglu-Zeybek AC,et al. Differences in the gut microbiota of healthy children and those withtype 1 diabetes. Pediatr Int. 2014;56:336-343. 30.Bekkering P, Jafri I, van Overveld FJ,et al. The intricate association between gut microbiota and development of type1, type 2 and type 3 diabetes. Expert Rev Clin Immunol. 2013;9:1031-1041. 31.Everard A, Cani PD. Diabetes, obesityand gut microbiota. Best Pract Res Clin Gastroenterol. 2013;27:73-83. 32.Everard A, Lazarevic V, Derrien M, etal. Responses of gut microbiota and glucose and lipid metabolism to prebioticsin genetic obese and diet-induced leptin-resistant mice. Diabetes.2011;60:2775-2786. 33.Lin CH, Lin CC, Shibu MA, et al. OralLactobacillus reuteri GMN-32 treatment reduces blood glucose concentrations andpromotes cardiac function in rats with streptozotocin-induced diabetesmellitus. Br J Nutr. 2014;111:598-605. 34.Yadav H, Jain S, Sinha PR. Effect ofskim milk and dahi (yogurt) on blood glucose, insulin, and lipid profile inrats fed with high fructose diet. J Med Food. 2006;9:328-335. 35.Andreasen AS, Larsen N,Pedersen-Skovsgaard T, et al. Effects of Lactobacillus acidophilus NCFM oninsulin sensitivity and the systemic inflammatory response in human subjects.Br J Nutr. 2010;104:1831-1838. 36.Shakeri H, Hadaegh H, Abedi F, et al.Consumption of synbiotic bread decreases triacylglycerol and VLDL levels whileincreasing HDL levels in serum from patients with type-2 diabetes. Lipids.2014;49:695-701. 37. Akbari V, Hendijani F, Effects of probiotic supplementation in patients with type 2 diabetes: systematic review and meta-analysis. Nutrition reviews, 2016; 74:274-284 38. Pfutzner A., Weber M.M., Forst T.: A Biomarker Concept for Assessment of Insulin resistance, β-Cell Function and Chronic Systemic Inflammation in Type 2 Diabetes mellitus. Clin Lab 2008; 54:485-490 39. Hollander P. Safety profile of acarbose, an alpha-glucosidase inhibitor. Drugs. 1992;44 Suppl 3:47-53. 40. Sels JP, Verdonk HE, Wolffenbuttel BH. Effects of acarbose (Glucobay) in persons with type 1 diabetes: a multicentre study. Diabetes Res Clin Pract. 1998 Aug;41(2):139-45. 4l. Kayibanda JF, Girouard C, Gregoire JP, Demers E, Moisan. Adherence to the evidence-based heart failure drug treatment: Are there sex-specific differences among new users? J. Res Social Adm Pharm. 2018;14:915-920

[0452]

Table 1

[0453] [Table 2]

[0454] The reader is directed to all articles and documents related to this application, filed contemporaneously or prior to this application, and open to public inspection herewith, the contents of which are incorporated herein by reference in their entirety.

[0455] All features disclosed in this specification (including all accompanying claims, abstracts, and drawings), and / or all steps of any disclosed methods or processes, may be combined in any combination, except combinations in which at least some of such features and / or steps are incompatible.

[0456] Each feature disclosed in this specification (including all accompanying claims, abstracts, and drawings), unless otherwise stated, may be replaced by alternative features serving the same, equivalent, or similar purpose. Thus, unless otherwise stated, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0457] The invention is not limited to the details of any of the above embodiments, and extends to any novel one or any novel combination of features disclosed herein (including all accompanying claims, abstracts, and drawings), or to any novel one or any novel combination of steps of any method or process disclosed.

Claims

1. A composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for use in treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject.

2. A method for treating and / or preventing type 2 diabetes, prediabetes, and / or symptoms thereof in a subject, comprising administering to the subject a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

3. 3. The composition used according to claim 1 or the method according to claim 2, wherein the composition is for the treatment and / or prevention of type 2 diabetes and / or its symptoms.

4. administering the composition to the subject (a) reducing the levels of one or more of HbA1c, glucose, insulin, intact proinsulin, and / or hsCRP in the blood; (b) reducing the HOMA-IR score, and / or (c) increasing blood adiponectin levels; 10. A composition or method for use according to any one of the preceding claims.

5. 10. A composition or method for use according to any one of the preceding claims, wherein administration of said composition to said subject controls blood glucose.

6. 10. A composition or method for use according to any one of the preceding claims, wherein administration of the composition to the subject degrades glucose.

7. 10. The composition or method of use of any one of the preceding claims, wherein administering the composition to the subject metabolizes glucose in the subject's digestive tract, optionally administering the composition to the subject metabolizes glucose in the subject's intestine, and further optionally administering the composition to the subject metabolizes glucose in the subject's small intestine.

8. 10. A composition or method for use according to any one of the preceding claims, wherein administration of the composition to the subject reduces glucose absorption into the blood of the subject.

9. 10. A composition or method for use according to any one of the preceding claims, wherein administration of said composition to said subject reduces blood glucose levels in said subject.

10. 10. A composition or method for use according to any one of the preceding claims, wherein the one or more Bacillus species is selected from B. subtilis and B. coagulans.

11. 11. The composition or method of use of claim 10, wherein the composition comprises B. subtilis and B. coagulans.

12. a) the B. subtilis species is selected from the group consisting of Bacillus subtilis strain DFM 0326 (LMG P-32899) and Bacillus subtilis strain DFM 1015 (LMG P-32900), and / or c) the B. coagulans species is Bacillus coagulans strain DFM 0705 (LMG P-32921); 12. A composition or method for use according to claim 10 or 11.

13. 10. A composition or method for use according to any one of the preceding claims, wherein the high molecular weight, low osmolality carbohydrate is a dextrin.

14. 10. The composition or method of use according to any one of the preceding claims, wherein the composition further comprises one or more bacterial species selected from the group consisting of Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus sp MT 03, Bacillus atrophaeus, and Pediococcus pentosaceus.

15. 10. A composition or method for use according to any one of the preceding claims, wherein the composition comprises at least about 20% w / w L-cysteine.

16. The composition is at least about 1×10 ５ 10. The composition or method of use according to any one of the preceding claims, comprising a bacterium of the genus Bacillus.

17. 10. A composition or method for use according to any one of the preceding claims, wherein the composition comprises at least about 73% w / w rice bran.

18. 10. A composition or method for use according to any one of the preceding claims, wherein the composition comprises at least about 0.5% w / w of high molecular weight, low osmolality carbohydrates.

19. 10. The composition or method of use according to any one of the preceding claims, wherein the composition further comprises one or more of vitamin B12, a magnesium salt of a fatty acid, calcium phosphate, potassium phosphate, silicon dioxide, and cellulose, and optionally the magnesium salt of a fatty acid is magnesium stearate.

20. 10. A composition or method for use according to any one of the preceding claims, wherein the composition is formulated as an acid-resistant tablet or capsule.

21. 21. The composition or method of use of claim 20, wherein the acid-resistant tablet or capsule comprises a film coating, the film coating comprising hydroxypropyl methylcellulose (HPMC).

22. 10. A composition or method for use according to any one of the preceding claims, wherein the one or more Bacillus species are not genetically modified.

23. 10. A composition or method for use according to any one of the preceding claims, wherein the composition is for daily administration.

24. Use of a composition comprising one or more species of Bacillus bacteria, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for degrading glucose.

25. (a) to reduce the levels of one or more of HbA1c, glucose, insulin, intact proinsulin, and / or hsCRP in the blood of a subject; (b) to reduce the HOMA-IR score in a subject; and / or (c) increasing blood adiponectin levels in a subject; Use of a composition comprising one or more species of Bacillus bacteria, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate.

26. Use of a composition comprising one or more Bacillus species, rice bran, L-cysteine, and a high molecular weight, low osmolality carbohydrate for controlling blood glucose in a subject.

27. a) the B. subtilis species is selected from the group consisting of Bacillus subtilis strain DFM 0326 (LMG P-32899) and Bacillus subtilis strain DFM 1015 (LMG P-32900), and / or c) the B. coagulans species is Bacillus coagulans strain DFM 0705 (LMG P-32921); Use according to any one of claims 24 to 26.

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