Methods and compositions for improving insulin production and secretion - Patents.com

Abstract

A method for improving insulin production in a subject in need thereof comprises administering to the subject a nutritional composition comprising lysine, arginine and β-hydroxy-β-methylbutyrate (HMB). A method for improving insulin secretion in a subject in need thereof comprises administering to the subject a nutritional composition comprising lysine, arginine and HMB. The nutritional composition comprises about 0.01 to about 15% by weight of HMB, about 0.03 to about 40% by weight of lysine, and about 0.02 to about 30% by weight of arginine.

Description

[Technical field]

[0001] The present invention relates to methods of improving insulin production in a subject, methods of improving insulin secretion in a subject, and nutritional compositions using lysine, arginine, and β-hydroxy-β-methylbutyrate (HMB). [Background technology]

[0002] Pancreatic β-cells produce and secrete insulin, a hormone that regulates blood glucose concentration. β-cells need to produce, store, and secrete insulin in sufficient concentrations to maintain normal glucose concentrations in the blood (i.e., euglycemia). Physiologically, euglycemia is governed by a balance between peripheral insulin sensitivity (how easily somatic cells in peripheral tissues such as muscle and fat can absorb glucose) and insulin secretion; when insulin sensitivity decreases, insulin secretion increases. Because β-cells are highly sensitive to blood glucose concentrations, changes in the body's ability to maintain blood glucose concentration equilibrium have a major impact on their function. With elevated blood glucose concentrations and prolonged insulin production, the pancreas loses the ability to adapt β-cell mass to insulin demand over time, thereby reducing functional β-cell mass. Dysregulation of functional β-cell mass is an important mechanistic factor associated with the development and progression of diabetes.

[0003] Although progress in diabetes treatment by pharmacotherapy continues, addressing diabetes progression is an important issue, since the etiology and mechanisms of diabetes development have yet to be fully elucidated. Two major problems exist: how to cure diabetes and how to reduce the prevalence of diabetes through primary and secondary prevention.

[0004] Insulin resistance and beta cell dysfunction play an important role in the development and progression of diabetes. Insulin resistance, which is often present many years before diabetes is diagnosed, reflects a reduced response to insulin in key target tissues such as muscle, liver and adipose tissue and has been shown to predict the development of the disease. Beta cell function is already reduced in patients with impaired glucose tolerance, such as those diagnosed with prediabetes, and is further reduced in those with type 2 diabetes. UKPDS studies have shown that beta cell function in subjects with type 2 diabetes may decline by 50% at the time of diagnosis and by 5% each year thereafter, suggesting that deterioration begins several years before onset. Lancet, 352 (1998): pp. 837-853 and pp. 854-865; Br Med J, 317 (1998), pp. 703-713 and pp. 713-720.

[0005] Because deficiency in functional β-cell mass is a necessary and early condition for the development of type 2 diabetes, methods that provide β-cell recovery and / or β-cell regeneration and maintain functional islet integrity are desirable for preventing, slowing the progression of, treating, ameliorating, and potentially curing type 2 diabetes.

[0006] Currently, the use of antidiabetic drugs is widespread. The available pharmacotherapies to treat diabetes have been developed as "symptomatic treatments" since they act mainly to reduce hyperglycemia levels. However, monotherapy with antidiabetic drugs (e.g., metformin, rosiglitazone, and glyburide) has been reported to fail over time, although at different rates of decline. Moreover, current treatments do not completely halt the progressive loss of beta-cell function, and their use is also associated with hypoglycemia and weight gain. To be able to prevent the onset and progression of type 2 diabetes, treatment should also halt beta-cell dysfunction and promote the restoration of fully functional beta-cell mass, independent of lowering hyperglycemia. Management of type 2 diabetes should ideally include early and simultaneous treatment of insulin resistance and beta-cell dysfunction.

[0007] As mentioned above, in addition to pharmacotherapy, significant efforts have also been made in understanding effective preventive management of both prediabetes and diabetes through exercise and nutritional interventions. Preventive strategies for management of diabetes progression, for example, the progression from prediabetes to diabetes, or from gestational diabetes to type 2 diabetes after pregnancy, should focus on improving insulin secretion and action, minimizing factors related to diabetes, especially insulin resistance, preventing hyperglycemia, and restoring muscle structure and function.

[0008] Therefore, methods for improving insulin secretion and production are desirable, especially for patients suffering from diabetes or prediabetes.It is also desirable to provide methods for improving insulin secretion and production that can help slow the progression of type 2 diabetes or prediabetes, prevent or reverse type 2 diabetes or type 2 diabetes.Nutrition interventions that can help address the above limitations associated with existing diabetes treatments are also desirable. Summary of the Invention

[0009] In one embodiment, the present invention relates to a method of improving insulin production in a subject in need thereof, comprising administering to the subject a nutritional composition comprising lysine, arginine and β-hydroxy-β-methylbutyrate (HMB).

[0010] In an additional embodiment, the present invention relates to a method of improving insulin secretion in a subject in need thereof, comprising administering to the subject a nutritional composition comprising lysine, arginine and HMB.

[0011] In yet an additional embodiment, the present invention relates to a nutritional composition comprising about 0.01 to about 15% by weight HMB, about 0.03 to about 40% by weight lysine, and about 0.02 to about 30% by weight arginine.

[0012] The method for improving insulin production and improving insulin secretion, as well as the nutritional composition according to the present invention, are advantageous in that they improve insulin secretion and insulin production, and thus may contribute to reducing β-cell degeneration and / or improving β-cell functionality. Thus, the method and nutritional composition of the present invention may be useful in delaying or preventing the onset of diabetes, delaying or preventing the progression of diabetes, and / or promoting remission. These and further objects and advantages of the present invention will become more fully apparent in view of the following detailed description.

[0013] The embodiments set forth in the drawings illustrate certain aspects of the invention and are exemplary in nature and are not intended to limit the invention as defined by the claims. [Brief description of the drawings]

[0014] [Figure 1] 1 shows the effect of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine and HMB (MIX) on glucose uptake in INS-1 cells, as described in Example 1. [Diagram 2] FIG. 1 shows the effects of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine, and HMB (MIX) on the expression levels of GLUT2, the main glucose transporter in pancreatic cells, in INS-1 cells, as described in Example 1. [Diagram 3] FIG. 1 shows the effects of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine, and HMB (MIX) on the acute response of insulin secretion to extracellular glucose concentration in INS-1 cells, as described in Example 2. [Figure 4] 1 shows the effect of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine and HMB (MIX) on insulin production in INS-1 cells, as described in Example 2. [Diagram 5]1 shows the effect of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine, and HMB (MIX) on intracellular insulin concentrations in INS-1 cells, as described in Example 2. [Figure 6] 1 shows the effects of arginine (ARG), lysine (LYS), HMB, and a combination of arginine, lysine and HMB (MIX) on beta cell proliferation and viability, as described in Example 3. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Specific embodiments of the present invention are described herein. However, the present invention may be embodied in different forms and should not be construed as being limited to the embodiments described herein. Rather, these embodiments are provided to illustrate more specific features of certain aspects of the present invention to those skilled in the art.

[0016] The terminology described herein is for the purpose of describing the embodiments only and should not be construed as limiting the entire disclosure. All references to singular features or limitations in this disclosure include the corresponding plural features or limitations, and vice versa, unless otherwise specified or clearly suggested to the contrary by the referenced context. Unless otherwise specified, "a", "an", "the", and "at least one" are used interchangeably. Furthermore, as used in this specification and the appended claims, the singular forms "a", "an", and "the" include their plural forms unless the context clearly indicates otherwise.

[0017] The term "includes" or "including", as used in the present specification or claims, is intended to encompass additional elements or steps in a similar manner to the term "comprise", as the term "comprising" is interpreted when used as a transitional term in a claim. Furthermore, to the extent the term "or" is used (e.g., A or B), it is intended to mean "A or B or both". When "A or B but not both" is intended, the term "A or B but not both" is used. Thus, the use of the term "or" herein is inclusive and not exclusive. When the terms "and" and "or" are used together, such as "A and / or B", this refers not only to A or B, but also to A and B.

[0018] The methods and compositions described in this disclosure can comprise, consist of, or consist essentially of any of the elements and steps described herein.

[0019] All ranges and parameters, including but not limited to percentages, portions, and ratios disclosed herein, are understood to encompass all subranges subsumed therein, and all numbers between the endpoints. For example, a range stated as "1 to 10" should be considered to include all subranges beginning with a minimum value of 1 or more and ending with a maximum value of 10 or less (e.g., 1 to 6.1, or 2.3 to 9.4), and each integer subsumed within that range (1, 2, 3, 4, 5, 6, 7, 8, 9, and 10).

[0020] Any combination of method or process steps used herein can be performed in any order unless otherwise specified or unless the contrary is clearly suggested by the context in which the combination is recited.

[0021] Unless otherwise indicated, all percentages are percentages by weight.

[0022] As used herein, the term "HMB" refers to β-hydroxy-β-methylbutyrate (also known as β-hydroxy-3-methylbutyric acid, β-hydroxyisovaleric acid) and sources thereof, unless otherwise specified. All weights, percentages and concentrations used herein to characterize HMB are based on the weight of HMB, regardless of source, unless otherwise specified.

[0023] As used herein, the term "calcium HMB" refers to the calcium salt of β-hydroxy-β-methylbutyrate (also known as β-hydroxy-3-methylbutyric acid, β-hydroxyisovaleric acid, or HMB), most commonly in the monohydrate form, unless otherwise specified. All weights, percentages, and concentrations used herein to characterize calcium HMB are based on the weight of calcium HMB monohydrate, unless otherwise specified.

[0024] The term "diabetes" as used herein refers to type 1 diabetes, type 2 diabetes, and gestational diabetes unless otherwise specified. Type 1 diabetes (also called juvenile diabetes or insulin-dependent diabetes) is a chronic disease in which the pancreas produces little or no insulin. It is considered an autoimmune condition because the immune system mistakenly attacks and destroys the insulin-producing beta cells in the pancreas. Type 2 diabetes is a chronic disease that affects the process by which the body metabolizes sugar. In type 2 diabetes patients, the body either resists the action of insulin, causing a decrease in glucose uptake by cells, or does not produce enough insulin to maintain normal glucose levels. Type 2 diabetes often begins as insulin resistance, which means the body cannot use insulin efficiently. Gestational diabetes occurs only during pregnancy and is the result of insulin-inhibiting hormones produced during pregnancy. Patients diagnosed with gestational diabetes are at high risk of developing type 2 diabetes after pregnancy.

[0025] The term "nutritional powder" as used herein, unless otherwise specified, refers to a nutritional powder that is generally a free-flowing particle, reconstitutable with an aqueous liquid, and suitable for oral administration to humans.

[0026] The term "nutritional liquid" as used herein, unless otherwise specified, refers to a nutritional product in ready-to-drink liquid form and refers to a nutritional liquid made by reconstituting the nutritional powder described herein prior to use.

[0027] The terms "nutritional product" and "nutritional composition" as used herein, unless otherwise specified, refer to nutritional liquids and nutritional powders, which may be reconstituted to form nutritional liquids, which are suitable for oral consumption by humans.

[0028] β-hydroxy-β-methylbutyrate (HMB) is a naturally occurring amino acid metabolite known for use in various nutritional products and supplements.HMB is a metabolite of the essential amino acid leucine, and has been shown to regulate protein turnover and inhibit protein degradation.Calcium HMB is the commonly used form of HMB when formulated into oral nutritional products, including tablets, capsules, reconstitutable powders, and nutritional liquids and emulsions.

[0029] While HMB is commonly used in nutritional products to help build or maintain healthy muscle in select individuals, the present inventors have surprisingly discovered that HMB, in combination with arginine and lysine, improves insulin secretion and production in pancreatic beta cells.

[0030] In one embodiment, a method for improving insulin production in a subject is provided. The method comprises administering to the subject a nutritional composition comprising lysine, arginine and HMB. In another embodiment of the present invention, a method for improving insulin secretion in a subject is provided. The method comprises administering to the subject a nutritional composition comprising lysine, arginine and HMB. In certain embodiments of these methods, the subject suffers from diabetes or prediabetes. In further embodiments of the methods of the present invention, the method further comprises administering to the subject a sugar alcohol. Suitable sugar alcohols include, but are not limited to, mannitol, sorbitol, xylitol, lactitol, isomalt, maltitol and myo-inositol. In a more specific embodiment, the sugar alcohol is myo-inositol.

[0031] In a further embodiment of the present invention, a nutritional composition is provided comprising HMB, lysine and arginine. In one embodiment, the nutritional composition comprises about 0.01 to about 15% by weight or about 0.01 to about 10% by weight of HMB, about 0.03 to about 40% by weight or about 0.03 to about 30% by weight of lysine, and about 0.02 to about 30% by weight or about 0.02 to about 20% by weight of arginine. In a specific embodiment of the nutritional composition of the present invention, the nutritional composition further comprises a sugar alcohol as described above. In a more specific embodiment, the sugar alcohol is myo-inositol.

[0032] In another embodiment of the nutritional composition of the present invention, the molar ratio of lysine to arginine in the nutritional composition is about 10:1 to about 1:1, or about 5:1 to about 1:1, or about 3:1 to about 1:1. In yet another particular embodiment, the molar ratio of the combination of lysine and arginine to HMB in the nutritional composition is about 15:1 to about 1:1, or about 10:1 to about 1:1, or about 5:1 to about 1:1, or about 3:1 to about 1:1.

[0033] Any source of HMB is suitable for use in the method and nutritional composition of the present invention.Examples include HMB as free acid, salt, including anhydrous salt or hydrated salt, ester, lactone, or other product form, which otherwise provides bioavailable form of HMB.In certain embodiments of the method and composition of the present invention, the HMB source is selected from the group consisting of alkali metal HMB, alkaline earth metal HMB, HMB free acid, HMB lactone, and combinations thereof.In more specific embodiments, the HMB source is selected from the group consisting of sodium HMB, potassium HMB, magnesium HMB, chromium HMB, calcium HMB, and combinations thereof, or HMB is calcium HMB monohydrate.

[0034] According to a specific embodiment of the method, the lysine, arginine and HMB are administered orally. In a more specific embodiment, the lysine, arginine and HMB are provided in a nutritional composition and administered orally.

[0035] The methods and nutritional compositions described herein improve insulin production and / or improve insulin secretion, and more specifically, use lysine, arginine and HMB in amounts effective to achieve one or more of these effects to a greater extent than is achieved by lysine, arginine or HMB alone.

[0036] In a particular embodiment, the nutritional composition is in the form of a powder. In another particular embodiment, the nutritional composition is in the form of a liquid.

[0037] In certain embodiments of the method, the subject is administered about 1 to about 10 g, or about 2 to about 5 g, of HMB per day.

[0038] In other specific embodiments of the method, the subject is administered about 0.1 g to about 30 g, or about 1 g to about 10 g, or about 3 g to about 6 g of lysine per day.

[0039] In further specific embodiments of the method, the subject is administered about 0.1 to about 20 g, or about 1 to about 10 g, or about 1 to about 5 g of arginine per day.

[0040] Lysine and arginine can be added to the nutritional composition either in their native form or in supplemented form. Native amino acids are those provided by food proteins, while supplemental amino acids are free amino acids in the L- or D-configuration. In a particular embodiment of the invention, the nutritional composition uses supplemental lysine and / or arginine. In a more specific embodiment of the invention, the nutritional composition uses lysine and / or arginine in the L-form.

[0041] In further specific embodiments of the present invention, the nutritional composition comprises about 0.01 to about 10 wt%, about 0.01 to about 8 wt%, about 0.01 to about 5.0 wt%, 0.1 to about 10 wt%, about 0.1 to about 8 wt%, about 0.1 to about 5.0 wt%, about 0.2 to about 5.0 wt%, about 0.3 to about 3 wt%, about 0.3 to about 2 wt%, about 0.3 to about 1.5 wt%, about 0.3 to about 1.0 wt%, about 0.3 to 0.6 wt%, or about 0.4 to about 1.5 wt% HMB based on the weight of the nutritional composition.

[0042] In another specific embodiment, the nutritional composition comprises about 0.03 to about 40% by weight, or about 0.03 to about 30% by weight, or about 0.03 to about 20% by weight of lysine based on the weight of the nutritional composition. In a more specific embodiment, the nutritional composition comprises about 0.1 to about 10% by weight of lysine based on the weight of the nutritional composition.

[0043] In another specific embodiment of the present invention, the nutritional composition comprises about 0.02 to about 30% by weight, or about 0.02 to about 20% by weight, or about 0.02 to about 10% by weight of arginine based on the weight of the nutritional composition. In a more specific embodiment, the nutritional composition comprises about 0.05 to about 5% by weight of arginine based on the weight of the nutritional composition.

[0044] In another specific embodiment of the present invention, the nutritional composition further comprises protein, carbohydrate, and / or fat in any amount as required.A wide variety of sources and types of protein, carbohydrate, and fat can be used in the nutritional composition embodiments described herein.In a specific embodiment, the nutritional composition comprises protein, carbohydrate, and fat.

[0045] In further particular embodiments, the protein of the nutritional composition is selected from the group consisting of whey protein concentrate, whey protein isolate, whey protein hydrolysate, milk protein concentrate, milk protein isolate, milk protein hydrolysate, milk protein concentrate, organic milk protein concentrate, soy protein concentrate, soy protein isolate, soy protein hydrolysate, pea protein concentrate, pea protein isolate, pea protein hydrolysate, acid casein, sodium caseinate, calcium caseinate, potassium caseinate, casein hydrolysate, skim milk powder, concentrated skim milk powder, cola The active ingredient in the composition of the present invention is omega-3 fatty acids, omega-3 fatty acids, glycerin, lecithin, lecithin, lecithin, lecithin fatty acids ...

[0046] In certain embodiments, the nutritional composition may comprise protein in an amount of about 1% to about 30% by weight of the nutritional composition. More specifically, the protein may be present in an amount of about 1% to about 25% by weight of the nutritional composition, including about 1% to about 20% by weight, about 2% to about 20% by weight, about 1% to about 15% by weight, about 1% to about 10% by weight, about 5% to about 10% by weight, about 10% to about 25% by weight, or about 10% to about 20% by weight. Even more specifically, the protein comprises about 1% to about 5% by weight of the nutritional composition, or about 20% to about 30% by weight of the nutritional composition.

[0047] In other particular embodiments, the carbohydrate of the nutritional composition is selected from the group consisting of fiber, human milk oligosaccharides (HMO), maltodextrin, corn maltodextrin, corn syrup, organic corn maltodextrin, corn syrup, corn syrup solids, sucralose, cellulose gel, cellulose gum, gellan gum, carrageenan, fructooligosaccharides (FOS), inositol, hydrolyzed starch, glucose polymers, carbohydrates derived from rice, sucrose, glucose, lactose, honey, sugar alcohols, isomaltulose, sucromalt, pullulan, potato denaturant, and the like. The carbohydrates in the nutritional composition include: bread, galactooligosaccharides, oat fiber, soybean fiber, corn fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, acacia gum, chitosan, arabinogalactin, glucomannan, xanthan gum, alginate, pectin, low methoxy pectin, high methoxy pectin, cereal beta glucan, psyllium, inulin, and combinations of two or more thereof. In further specific embodiments, the carbohydrates in the nutritional composition include combinations of two or more carbohydrates, and the carbohydrates have different absorption rates. In certain embodiments, the carbohydrates that can be used in the nutritional composition of the present invention include isomaltulose, sucromalt, resistant maltodextrin (e.g., fibersol or nutriose), FOS, inulin, oat fiber, soybean fiber, or combinations of two or more thereof.

[0048] In certain embodiments, the nutritional composition may contain carbohydrates in an amount of about 0.5% to about 75% by weight of the nutritional composition. More specifically, carbohydrates may be present in an amount of about 1% to about 70% by weight of the nutritional composition, including about 5% to about 70% by weight, about 5% to about 65% by weight, about 5% to about 50% by weight, about 5% to about 40% by weight, about 5% to about 30% by weight, about 5% to about 25% by weight, about 10% to about 65% by weight, about 20% to about 65% by weight, about 30% to about 65% by weight, about 40% to about 65% by weight, about 40% to about 70% by weight, or about 15% to about 25% by weight of the nutritional composition.

[0049] As used herein, the terms "fat" and "oil" are used interchangeably to refer to lipid materials derived or processed from plants or animals, unless otherwise specified. These terms also include synthetic lipid materials, so long as such synthetic materials are suitable for oral administration to humans.

[0050] In further specific embodiments, the fat comprises one or more fatty acids such as coconut oil, fractionated coconut oil, soybean oil, soybean lecithin, corn oil, safflower oil, sunflower oil, palm olein, canola oil, monoglycerides, lecithin, canola oil, medium chain triglycerides, linoleic acid, alpha-linoleic acid, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, medium chain triglyceride oil (MCT oil), high gamma linoleic acid (GLA), safflower oil, sunflower oil, palm oil, palm kernel oil, marine oil, fish oil, algae oil, borage oil, cottonseed oil, fungal oil, interesterified oil, transesterified oil, structured lipids, omega-3 fatty acids, or a combination of two or more thereof. In certain embodiments, the omega-3 fatty acid is selected from the group consisting of eicosapentaenoic acid, docosahexaenoic acid, arachidonic acid, and alpha-linoleic acid, and combinations of two or more thereof.

[0051] In certain embodiments, the nutritional composition may include fat in an amount of about 0.5% to about 30% by weight of the nutritional composition. More specifically, fat may be present in an amount of about 0.5% to about 10% by weight, about 1% to about 30% by weight of the nutritional composition, including about 1% to about 20% by weight, about 1% to about 15% by weight, about 1% to about 10% by weight, about 1% to about 5% by weight, about 3% to about 30% by weight, about 5% to about 30% by weight, about 5% to about 25% by weight, about 5% to about 20% by weight, about 5% to about 10% by weight, or about 10% to about 20% by weight of the nutritional composition.

[0052] The concentrations and relative amounts of protein, carbohydrate, and fat sources in the exemplary nutritional compositions may vary considerably, depending, for example, on the particular dietary needs of the intended user. In a particular embodiment, the nutritional composition comprises a protein source in an amount of about 2% to about 20% by weight, a carbohydrate source in an amount of about 5% to about 30% by weight, and a fat source in an amount of about 0.5% to about 10% by weight, based on the weight of the nutritional composition, more particularly, such compositions are in liquid form. In another particular embodiment, the nutritional composition comprises a protein source in an amount of about 10% to about 25% by weight, a carbohydrate source in an amount of about 40% to about 70% by weight, and a fat source in an amount of about 5% to about 20% by weight, based on the weight of the nutritional composition, more particularly, such compositions are in powder form.

[0053] In one embodiment, the nutritional composition is a liquid nutritional composition and comprises, based on the weight of the nutritional composition, about 1 to about 15% by weight protein, about 0.5 to about 10% by weight fat, and about 5 to about 30% by weight carbohydrate.

[0054] In another embodiment, the nutritional composition is a nutritional powder composition and comprises, based on the weight of the nutritional composition, about 10 to about 30% by weight protein, about 5 to about 15% by weight fat, and about 30 to about 65% by weight carbohydrate.

[0055] In certain embodiments, the nutritional composition comprises at least one protein comprising milk protein concentrate and / or soy protein isolate, at least one fat comprising canola oil, corn oil, coconut oil and / or fish oil, and at least one carbohydrate comprising maltodextrin, resistant maltodextrin, sucrose and / or short chain fructooligosaccharides.

[0056] The nutritional composition may also include one or more ingredients to modify the physical, chemical, aesthetic or processing properties of the nutritional composition, or to function as additional nutritional ingredients. Non-limiting examples of additional ingredients include preservatives, emulsifiers (e.g., lecithin), buffers, sweeteners, including artificial sweeteners (e.g., saccharin, aspartame, acesulfame K, sucralose), colorants, flavorings, thickeners, stabilizers, etc.

[0057] In a particular embodiment, the nutritional composition has a neutral pH, i.e., a pH of about 6 to 8, more particularly about 6 to 7.5, In a more particular embodiment, the nutritional composition has a pH of about 6.5 to 7.2, or more particularly about 6.8 to 7.1.

[0058] Nutritional composition can be formed by any technique known in the art.In one embodiment, nutritional composition can be formed by: (a) preparing an aqueous solution containing protein and carbohydrate; (b) preparing an oil blend containing fat and oil-soluble components; and (c) mixing the aqueous solution and the oil blend to form an emulsified liquid nutritional composition.HMB, lysine and arginine can be added at any time during the formation of nutritional composition.

[0059] As noted above, the nutritional composition may be administered in powder form or in liquid form.

[0060] When the nutritional composition is a powder, for example, the dosage is about 40 g to about 60 g, e.g., 45 g, or 48.6 g, or 50 g, and is administered as a powder or reconstituted in about 1 ml to about 500 ml of liquid.

[0061] When the nutritional composition is in liquid form, for example reconstituted from a powder or manufactured as a ready-to-drink product, the serving size ranges from about 1 ml to about 500 ml, including about 110 ml to about 500 ml, about 110 ml to about 417 ml, about 120 ml to about 500 ml, about 120 ml to about 417 ml, about 177 ml to about 417 ml, about 207 ml to about 296 ml, about 230 ml to about 245 ml, about 110 ml to about 237 ml, about 120 ml to about 245 ml, about 110 ml to about 150 ml, and about 120 ml to about 150 ml. In certain embodiments, the serving size is about 1 ml, or about 100 ml, or about 225 ml, or about 237 ml, or about 500 ml.

[0062] In certain embodiments, the nutritional composition comprising HMB, lysine and arginine is administered to the subject one or more times per day, or one or more times per week. In certain embodiments, the nutritional composition is administered to the subject about once to about six times per day or week, or about once to about five times per day or week, or about once to about four times per day or week, or about once to about three times per day or week. In certain embodiments, the nutritional composition is administered once or twice per day for at least one week, at least two weeks, at least three weeks, or at least four weeks.

[0063] In another embodiment of the present invention, the nutritional composition comprises one or more nutrients selected from the group consisting of proteins, carbohydrates, fats, and vitamins and minerals. Particular embodiments of the nutritional composition may include vitamins and / or related nutrients, non-limiting examples of which include vitamin A, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, niacin, folic acid, pantothenic acid, biotin, choline, inositol, and / or salts and derivatives thereof, and combinations thereof.

[0064] Certain embodiments of the nutritional composition include minerals, non-limiting examples of which include calcium, phosphorus, magnesium, zinc, manganese, sodium, potassium, molybdenum, chromium, iron, copper, and / or chloride, and combinations thereof.

[0065] The following examples demonstrate aspects of the method of the present invention and are provided for illustrative purposes only. The examples should not be construed as limiting the general inventive concept, many variations of which are possible without departing from the spirit and scope of the general inventive concept. EXAMPLES

[0066] Example 1: Effects of HMB, lysine and arginine on glucose uptake in a rat insulinoma cell line (INS-1) This example describes the use of a rat insulinoma cell line (INS-1) capable of releasing insulin in response to glucose stimulation to detect changes in glucose metabolism and insulin secretion, as well as the expression levels of the pancreatic glucose transporter GLUT2.

[0067] INS-1 cells were maintained in RPMI 1640 medium supplemented with 11.1 mmol / L D-glucose, 10% fetal bovine serum, 100 units / mL penicillin, 100 μg / mL streptomycin, 10 mmol / L HEPES, 2 mmol / L L-glutamine, 1 mmol / L sodium pyruvate, and 50 μmol / L β-mercaptoethanol in a humidified atmosphere at 37°C / 5% CO2.

[0068] The cells were incubated overnight with the different effectors. The following cell populations were used: (i) Control: INS-1 cells without components. (ii) ARG: INS-1 cells incubated in the presence of 5 mM arginine. (iii) LYS: INS-1 cells incubated in the presence of 10 mM lysine. (iv) HMB: INS-1 cells incubated in the presence of 25 μM HMB free acid. (v) MIX: INS-1 cells incubated in the presence of a combination of 5 mM arginine, 10 mM lysine and 25 μM HMB.

[0069] Uptake of 2-deoxy-[3H]d-glucose (2-DG) (10 μmol / L) was measured over 10 min under conditions where uptake was proportional. 2-DG uptake was terminated after 10 min by rapid aspiration of the radioactive incubation medium and washing three times in ice-cold phosphate-buffered saline. Cell-associated radioactivity was determined by cell lysis in 0.5 N NaOH with neutralization by addition of 0.5 N HCl followed by liquid scintillation. Protein concentration was measured using an aliquot from each well using the bicinchoninic acid (BCA) protein assay.

[0070] As shown in Figure 1, INS-1 cells were incubated with the individual amino acids, lysine and arginine, resulting in increased glucose uptake. Lysine (10 mM) increased glucose uptake by approximately 100% compared to the control, arginine (5 mM) increased glucose uptake by approximately 52% compared to the control, and HMB increased glucose uptake by approximately 31% compared to the control. However, when cells were incubated with a combination of lysine (10 mM), arginine (5 mM) and HMB (25 μM), glucose uptake was significantly increased. As shown in Figure 1, cells incubated with a combination of both amino acids and HMB showed an increase in glucose uptake of approximately 162% compared to the control and an increase in glucose uptake of approximately 31% compared to lysine alone. These results demonstrate that the combination of lysine, arginine and HMB exhibits a surprising synergistic effect on glucose uptake in β cells.

[0071] FIG. 2 shows the effect of lysine (10 mM), arginine (5 mM), HMB (25 μM) and their combination on the expression level of the main glucose transporter, GLUT2, in pancreatic cells. Incubation with lysine (10 mM) alone increased the relative intensity of the expression level of GLUT2 by about 90% compared to the control, and incubation with arginine (5 mM) increased the relative intensity of the expression level of GLUT2 by about 30% compared to the control. However, incubation with HMB (25 μM) alone decreased the relative intensity of the expression level of GLUT2 by about 23% compared to the control, while the combination of lysine (10 mM), arginine (5 mM) and HMB (25 μM) showed a surprising synergistic effect on glucose uptake. Incubation of cells with a combination of lysine (10 mM), arginine (5 mM) and HMB (25 μM) increased the relative intensity of the expression level of GLUT2 by about 220% compared to the control.

[0072] These results indicate that the combination of arginine, lysine and HMB exhibits a synergistic effect on glucose uptake in INS-1 cells.

[0073] Example 2: Effects of HMB, lysine and arginine on insulin secretion and insulin production in a rat insulinoma cell line (INS-1) This example describes the use of a rat insulinoma cell line to detect acute insulin secretion and insulin production.

[0074] Acute insulin secretion was measured by incubating INS-1 cells for 2 hours in the presence of the following effectors: glucose (10 mM), and the combination of the effectors arginine (5 mM), lysine (10 mM), HMB (25 μM), and arginine (5 mM), lysine (10 mM) and HMB (25 μM). INS-1 cells without effectors were used as controls. Insulin was detected using a Rat Insulin Enzyme Immunoassay (RIEI) kit (Mercodia, Sweden) according to the manufacturer's instructions. As shown in Figure 3, the acute response of insulin secretion to extracellular glucose concentration in the individual presence of arginine, lysine and HMB showed no significant difference with respect to the control. However, the combination of arginine, lysine and HMB significantly increased insulin secretion by about 79% compared to the control. This result demonstrates that the combination of arginine, lysine and HMB shows a surprising synergistic effect on insulin secretion in β-cells.

[0075] Insulin production by β-cells was assessed by incubating INS-1 cells in the presence of the following effectors: arginine (5 mM), lysine (10 mM), HMB (25 μM), and a combination of arginine (5 mM), lysine (10 mM) and HMB (25 μM). INS-1 cells without effectors were again used as controls. Cells were incubated with each of the above effectors for 24 hours. Following the initial 24-hour incubation period with the effectors, the same INS-1 cells were incubated with glucose (10 mM) for 2 hours. Insulin was again detected using a RIEI kit according to the manufacturer's instructions.

[0076] As shown in Figure 4, insulin production in cells incubated with the individual effectors arginine (5 mM), lysine (10 mM) and HMB (25 μM) was slightly increased compared to the control, but surprisingly, insulin production was 207% in INS-1 cells incubated with the combination of arginine (5 mM), lysine (10 mM) and HMB (25 μM). These results indicate that the combination of arginine (5 mM), lysine (10 mM) and HMB (25 μM) exhibits a surprising synergistic effect on promoting insulin production in pancreatic β cells.

[0077] The above-mentioned effects of increasing insulin secretion and production in INS-1 cells were confirmed by incubating INS-1 cells with a combination of arginine (5 mM), lysine (10 mM) and HMB (25 μM) for 24 hours with each of the above-mentioned effectors, followed by cell lysis. Protein concentration was measured using a BCA assay. As shown in Figure 5, intracellular insulin levels also demonstrated the synergistic effect observed with the combination of arginine, lysine and HMB by significantly increasing insulin production compared to both the control and the individual effectors.

[0078] Example 3: Effect of HMB, lysine and arginine on preserving β-cell mass in a rat insulinoma cell line (INS-1) This example describes the use of a rat insulinoma cell line (INS-1) to detect the effect of a combination of lysine, arginine and HMB in the presence of free fatty acids (e.g. palmitic acid) on the functional properties of β-cells using a standardized protocol for viability. In type 2 diabetes patients, chronic increase in plasma free fatty acid concentration leads to lipid metabolism dysregulation, which contributes to reduced β-cell function and viability (lipotoxicity). INS-1 cells were maintained in RPMI1640 medium supplemented with 11.1 mmol / L D-glucose, 10% fetal bovine serum, 100 units / mL penicillin, 100 μg / mL streptomycin, 10 mmol / L HEPES, 2 mmol / L L-glutamine, 1 mmol / L sodium pyruvate and 50 μmol / L β-mercaptoethanol at 37°C / 5% CO2 in a humidified atmosphere.

[0079] Preaggregated pancreatic cells were left untreated (control) or treated with palmitate at a concentration of 250 μM for 24 h in complete medium (palmitate). Four different effectors were added 3 h prior to incubation with palmitate: arginine (5 mM), lysine (10 mM), HMB (25 μM), and a combination of arginine (5 mM), lysine (10 mM) and HMB (25 μM).

[0080] The cell populations used in this experiment were as follows: (i) Control: INS-1 cells without components. (ii) Palmitate: INS-1 cells incubated in the presence of 250 μM palmitate. (iii) Palmitate+ARG: INS-1 cells incubated in the presence of 250 μM palmitate and 5 mM arginine. (iv) Palmitate+LYS: INS-1 cells incubated in the presence of 250 μM palmitate and 10 mM lysine. (v) Palmitate + HMB: INS-1 cells incubated in the presence of 250 μM palmitate and 25 μM HMB free acid. (vi) Palmitate+MIX: INS-1 cells incubated in the presence of 250 μM palmitate and a combination of 5 mM arginine, 10 mM lysine and 25 μM HMB.

[0081] Cell viability assays were performed by indirect measurement of cell metabolic activity using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). After 24 h of incubation with the above effectors, MTT solution (0.5 mg / ml) was added for 30 min. The supernatant was removed, 100 μl of MTT solvent was added to each well, and the culture plate was shaken for 10 min, after which the optical density (OD) values ​​were recorded at 570 nm.

[0082] As shown in Figure 6, cell viability at 24 hours was negatively affected by palmitate incubation. Data are presented as mean ± SEM. A p-value <0.05 versus control (*) or Mix (#) was considered significant. Incubation with individual amino acids and HMB did not show a favorable effect on the exposure of cells to palmitate. In fact, incubation with arginine, lysine and HMB individually reduced cell viability compared to treatment with palmitate alone. However, the combination of lysine, arginine and HMB (i.e. Mix) prevented the adverse effect of palmitate on cell viability and showed significantly higher cell proliferation compared to the remaining groups treated with palmitate. This increase was approximately 20% higher than cells incubated with palmitate alone. These results indicate that the combination of lysine, arginine and HMB shows a synergistic effect on improving beta cell mass and therefore functionality.

[0083] The results of these examples demonstrate the surprising synergistic effect of the combination of lysine, arginine and HMB to enhance insulin production and secretion in pancreatic β-cells. These results indicate that administration of the combination of lysine, arginine and HMB improves β-cell function, which is particularly relevant for the treatment of diabetes and prediabetes, and for the prevention of the progression of diabetes and its related diseases.

[0084] Although the present invention has been illustrated by the description of its embodiments and the embodiments have been described in considerable detail, such description is not intended to restrict or in any way limit the scope of the appended claims to such details. Additional advantages and modifications will be readily apparent to those skilled in the art. Thus, the present invention in its broader aspects is not limited to the specific details, representative compositions and methods, or illustrative examples shown and described. Accordingly, departures may be made from such details without departing from the spirit or scope of the general inventive concept.

Claims

1. A nutritional composition for improving insulin production in a subject in need thereof, comprising lysine, arginine and beta-hydroxy-beta-methylbutyrate (HMB).

2. A nutrient composition for improving insulin secretion in a subject in need thereof comprising lysine, arginine and beta-hydroxy-beta-methylbutyrate (HMB).

3. The nutritional composition of claim 1 or 2, wherein the subject is suffering from diabetes or prediabetes.

4. The nutritional composition according to any one of claims 1 to 3, wherein the composition comprises a sugar alcohol or myo-inositol.

5. 5. The nutritional composition of any one of claims 1 to 4, wherein the HMB is selected from the group consisting of alkali metal HMB, alkaline earth metal HMB, HMB free acid, HMB lactone, and combinations thereof, or the HMB is selected from the group consisting of sodium HMB, potassium HMB, magnesium HMB, chromium HMB, calcium HMB, and combinations thereof, or the HMB is calcium HMB monohydrate.

6. 6. The nutritional composition of any one of claims 1 to 5, wherein the molar ratio of lysine to arginine in the nutritional composition is from about 10:1 to about 1:1, or from about 5:1 to about 1:1, or from about 3:1 to about 1:1, and the molar ratio of a combination of lysine and arginine to HMB in the nutritional composition is from about 15:1 to about 1:1, or from about 10:1 to about 1:1, or from about 5:1 to about 1:

1.

7. The nutritional composition according to any one of claims 1 to 6, wherein the nutritional composition is administered orally to the subject.

8. The nutritional composition according to any one of claims 1 to 7, wherein the nutritional composition is in the form of a powder.

9. The nutritional composition according to any one of claims 1 to 7, wherein the nutritional composition is in liquid form.

10. providing the subject with about 1 to about 10 g, or about 2 to about 5 g, of HMB per day; and / or about 0.1 to about 30 g, about 1 to about 10 g, or about 4 to about 5 g of lysine per day; and / or 10. The nutritional composition of any one of claims 1 to 9, wherein the composition provides from about 0.1 to about 20 g, from about 1 to about 10 g, or from about 2 to about 3 g of arginine per day.

11. The nutritional composition according to any one of claims 1 to 10, further comprising protein, carbohydrate and / or fat.

12. The protein may be whey protein concentrate, whey protein isolate, whey protein hydrolysate, milk protein concentrate, milk protein isolate, milk protein hydrolysate, milk protein concentrate, organic milk protein concentrate, soy protein concentrate, soy protein isolate, soy protein hydrolysate, pea protein concentrate, pea protein isolate, pea protein hydrolysate, acid casein, sodium caseinate, calcium caseinate, potassium caseinate, casein hydrolysate, skim milk powder, concentrated skim milk powder, collagen protein, collagen tan.

12. The nutritional composition of claim 11 comprising: protein isolate, L-carnitine, taurine, lutein, rice protein concentrate, rice protein isolate, rice protein hydrolysate, faba bean protein concentrate, faba bean protein isolate, faba bean protein hydrolysate, meat protein, potato protein, chickpea protein, canola protein, mung protein, keana protein, amaranth protein, chia protein, hemp protein, flaxseed protein, earthworm protein, insect protein, or a combination of two or more thereof.

13. The carbohydrates include fiber, human milk oligosaccharides (HMO), maltodextrin, resistant maltodextrin, corn maltodextrin, corn syrup, organic corn maltodextrin, corn syrup, corn syrup solids, sucralose, cellulose gel, cellulose gum, gellan gum, carrageenan, fructooligosaccharides (FOS), inositol, hydrolyzed starch, glucose polymers, carbohydrates derived from rice, sucrose, glucose, lactose, honey, isomaltulose, sucromalt, pullulan, potato starch, galactooligosaccharides, oat fiber.

13. The nutritional composition of claim 11 or claim 12, comprising an ingredient selected from the group consisting of fiber, soy fiber, corn fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, acacia gum, chitosan, arabinogalactin, glucomannan, xanthan gum, alginates, pectin, low methoxy pectin, high methoxy pectin, cereal beta glucan, psyllium, inulin, and combinations of two or more thereof.

14. 14. The nutritional composition of claim 13, wherein the carbohydrate comprises isomaltulose, sucromalt, maltodextrin, resistant maltodextrin, FOS, inulin, oat fiber, soy fiber, or a combination of two or more thereof.

15. The fats include coconut oil, fractionated coconut oil, soybean oil, soybean lecithin, corn oil, safflower oil, sunflower oil, palm olein, canola oil, monoglycerides, lecithin, canola oil, medium chain triglycerides, linoleic acid, alpha-linoleic acid, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, medium chain triglyceride oil (MCT oil), high gamma linoleic acid (GLA), safflower oil, sunflower oil, palm oil, palm kernel oil, marine oil, fish oil, algae oil, borage oil, cottonseed oil, fungal oil, interesterified oil, transesterified oil, and the like.

15. The nutritional composition of any one of claims 11 to 14, comprising a lipid component selected from the group consisting of fatty acids, omega-3 fatty acids, structured lipids, and the like.

16. The nutritional composition according to any one of claims 11 to 15, wherein the nutritional composition comprises one or more nutrients selected from the group consisting of proteins, carbohydrates, fats, and vitamins and minerals.

17. 1. A nutritional composition comprising: From about 0.01 to about 15% by weight HMB; From about 0.03 to about 40% by weight of lysine; and The nutritional composition comprising from about 0.02 to about 30% arginine by weight, all based on the weight of the nutritional composition.

18. The nutritional composition comprises: about 0.01 to about 10%, about 0.01 to about 8%, about 0.01 to about 5.0%, about 0.1 to about 10%, about 0.1 to about 8%, about 0.1 to about 5.0%, about 0.2 to about 5.0%, about 0.3 to about 3%, about 0.3 to about 2%, about 0.3 to about 1.5%, about 0.3 to about 1.0%, about 0.3 to 0.6%, or about 0.4 to about 1.5% by weight HMB; about 0.03 to about 30%, about 0.03 to about 20%, or about 0.1 to about 10% by weight of lysine; and 18. The nutritional composition of claim 17, comprising from about 0.02 to about 20%, from about 0.02 to about 10%, or from about 0.05 to about 5% arginine by weight, all based on the weight of the nutritional composition.

19. 19. The nutritional composition of claim 17 or 18, wherein the nutritional composition further comprises a sugar alcohol or further comprises myo-inositol.

20. The nutritional composition according to any one of claims 17 to 19, further comprising protein, carbohydrate and / or fat.

21. The protein may be whey protein concentrate, whey protein isolate, whey protein hydrolysate, milk protein concentrate, milk protein isolate, milk protein hydrolysate, milk protein concentrate, organic milk protein concentrate, soy protein concentrate, soy protein isolate, soy protein hydrolysate, pea protein concentrate, pea protein isolate, pea protein hydrolysate, acid casein, sodium caseinate, calcium caseinate, potassium caseinate, casein hydrolysate, skim milk powder, concentrated skim milk powder, collagen protein, collagen protein isolate, L-carnitine, L 21. The nutritional composition of claim 20 comprising lysine, taurine, lutein, rice protein concentrate, rice protein isolate, rice protein hydrolysate, faba bean protein concentrate, faba bean protein isolate, faba bean protein hydrolysate, collagen protein, collagen protein isolate, meat protein, potato protein, chickpea protein, canola protein, mung protein, kean protein, amaranth protein, chia protein, hemp protein, flaxseed protein, earthworm protein, insect protein, or a combination of two or more thereof.

22. The carbohydrates include fiber, human milk oligosaccharides (HMO), maltodextrin, resistant maltodextrin, corn maltodextrin, corn syrup, organic corn maltodextrin, corn syrup, corn syrup solids, sucralose, cellulose gel, cellulose gum, gellan gum, carrageenan, fructooligosaccharides, inositol, hydrolyzed starch, glucose polymers, carbohydrates derived from rice, sucrose, glucose, lactose, honey, isomaltulose, sucromalt, pullulan, potato starch, galactooligosaccharides, oat fiber, 22. The nutritional composition of claim 20 or claim 21 comprising soy fiber, corn fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, acacia gum, chitosan, arabinogalactin, glucomannan, xanthan gum, alginates, pectin, low methoxy pectin, high methoxy pectin, cereal beta glucan, psyllium, inulin, and combinations of two or more thereof.

23. 23. The nutritional composition of claim 22, wherein the carbohydrate comprises isomaltulose, sucromalt, maltodextrin, resistant maltodextrin, FOS, inulin, oat fiber, soy fiber, or a combination of two or more thereof.

24. The fat may be one or more fatty acids such as coconut oil, fractionated coconut oil, soybean oil, soybean lecithin, corn oil, safflower oil, sunflower oil, palm olein, canola oil, monoglycerides, lecithin, canola oil, medium chain triglycerides, linoleic acid, alpha-linoleic acid, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, medium chain triglyceride oil (MCT oil), high gamma linoleic acid (GLA), safflower oil, sunflower oil, palm oil, palm kernel oil, marine oil, fish oil, algae oil, borage oil, cottonseed oil, fungal oil, interesterified oil, transesterified oil, or the like.

24. The nutritional composition of any one of claims 17 to 23, comprising a lipid component selected from the group consisting of fatty acids, omega-3 fatty acids, structured lipids, and the like.

25. The nutritional composition according to any one of claims 17 to 24, wherein the nutritional composition comprises one or more nutrients selected from the group consisting of proteins, carbohydrates, fats, and vitamins and minerals.

26. 26. The nutritional composition of any one of claims 17 to 25, wherein the nutritional composition comprises about 1 to about 15% by weight protein, about 0.5 to about 10% by weight fat, and about 1 to about 20% by weight carbohydrate, based on the weight of the nutritional composition.

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