Agglomerates of soluble whey protein aggregates and medical uses thereof

Abstract

The present invention relates to agglomerates of soluble whey protein aggregates for use in the treatment and / or prevention of one or more conditions associated with GLP-1 blood levels in a human subject.

Description

Field of the Invention

[0001] The present invention relates to agglomerates of soluble whey protein aggregates for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in human subjects. Background of the Invention

[0003] GLP-1 is known to affect a large number of biological processes in the human body.

[0004] Over the past few decades, the prevalence of overweight and obesity has increased significantly worldwide, a situation that imposes enormous costs on individuals and society in terms of both economics and health. Obesity develops when energy intake is greater than energy expenditure, and the excess energy is stored primarily as fat in adipose tissue. During the development of obesity, lipids also accumulate in non-adipose tissues, leading to further metabolic deterioration, such as inflammation and decreased insulin sensitivity.

[0005] Weight loss and prevention of weight gain can be achieved by reducing energy intake or bioavailability, increasing energy expenditure and / or reducing fat storage. There is a continuous need in the food industry to find better nutritional solutions to combat overweight and obesity.

[0006] Intestinal GLP-1 is an endogenous satiety signal; GLP-1 is secreted in response to food intake; intestinal GLP-1 secretion occurs within the time frame of meals; peripheral injection of physiological doses of GLP-1 reduces meal size; antagonizing endogenous GLP-1 increases meal size (Krieger, 2020).

[0007] Intestinal GLP-1 most likely reduces food intake via the GLP-1R (R = receptor) present on vagal afferents (Krieger, 2020). However, low circulating concentrations of GLP-1 after a meal do not automatically exclude the possibility of an endocrine effect of intestinal GLP-1. This is supported by the finding that GLP-1 induces its incretin effect in an endocrine manner primarily via GLP-1R activation in the pancreas. In relation to the effect of GLP-1 on feeding, concentrations of GLP-1 may be sufficient to activate GLP-1R in brain areas located outside the blood-brain barrier. The findings also indicate that endogenous GLP-1 of intestinal origin induces satiety via the vagus nerve rather than the central GLP-1R (Krieger, 2020).

[0008] Treatment with GLP-1RA (RA=receptor agonist) promotes weight loss by reducing appetite / food cravings and enhancing satiety, delaying gastric emptying, and through mechanisms that modulate gut-to-brain communication, in (Patel Chavez et al., 2022).

[0009] The global prevalence of type 2 diabetes has similarly increased significantly in both adults and children. Some of the increase may be due to lifestyle factors, similar to the rise in obesity rates due to changes in dietary patterns. There is a continuing need in the food industry to further improve nutritional solutions provided to diabetic subjects or subjects at risk of developing type 2 diabetes. In particular, to improve the insulin and glucagon profiles in diabetic or prediabetic subjects.

[0010] It is well known that incretins, glucagon-like peptide-1 (GLP-1), and glucose-dependent insulinotropic polypeptide (GIP) have pleiotropic effects on a variety of tissues including pancreatic beta cells, liver, arteries, heart, neurons, and adipose tissue. Although there are multiple GLP-1 target tissues, GLP-1 plays a vital role in pancreatic beta cells; GLP-1 binds to the GLP-1 receptor (GLP-1R) in the beta cell membrane, thereby enhancing insulin secretion, inhibiting apoptotic cell death, and increasing beta cell proliferation. In addition, GLP-1 inhibits glucagon secretion (from pancreatic alpha cells). In summary, this leads to improved blood sugar control (Kaneto et al., 2021).

[0011] In response to food intake, GLP-1 and GIP are secreted from the gastrointestinal tract and stimulate the secretion of insulin from pancreatic beta cells. Both incretin hormones bind to their respective receptors in the beta cell membrane, which leads to enhanced insulin secretion, reduced beta cell apoptosis and promoted beta cell proliferation. However, this effect of incretin hormones is significantly reduced in diabetic conditions in humans and rodents such as mice and rats. It is reported that in diabetic conditions, the expression level of incretin receptors is reduced, which may be related to the development of impaired incretin action and beta cell failure found in T2DM (Kaneto et al., 2021).

[0012] Insulin secretion is regulated by various intracellular signals in β-cells. Kaneto et al., 2021 reported that circulating physiological concentrations of GLP-1 directly stimulate insulin secretion from pancreatic β-cells. GLP-1 stimulates insulin secretion in a PKC-dependent or PKA-dependent manner depending on its concentration.

[0013] In addition to its effects on body weight and glucose, GLP-1 can also regulate blood cholesterol and triglycerides in many ways. Acute and long-term treatment with GLP-1 or its stable analogs reduces fasting as well as postprandial lipids in healthy and T2DM patients (Patel et al., 2014).

[0014] Another increasingly common global health issue is the development of non-alcoholic fatty liver disease, which leads to chronic liver disease through the accumulation of fat in the liver. There is a constant need within the food industry to find better nutritional solutions to combat the occurrence of this condition.

[0015] In a recent RCT including patients with NAFLD, the GLP-1RA semaglutide significantly reduced hepatic steatosis compared with placebo, which, together with improvements in liver enzymes and metabolic parameters, indicated a positive effect on disease activity and metabolic profile. Their results showed that semaglutide reduced fat content in hepatocytes, suggesting the potential to reverse steatosis and prevent the progression of fibrosis (Flint et al., 2021). In short, it is likely that in the near future, GLP-1RA will play a greater role in clinicians' treatment of patients with NAFLD (Patel Chavez et al., 2022).

[0016] Another global health-related issue is the occurrence of cardiovascular events, which is one of the leading causes of death worldwide. Heart attacks and strokes are usually acute events and are mainly caused by blockages that prevent blood flow to the heart or brain. There is a continuous need in the food industry to find better nutritional solutions to combat the occurrence of cardiovascular events.

[0017] GLP-1 receptors are expressed in endothelial cells and smooth muscle cells. In endothelial cells, incretin signaling improves vascular relaxation responses through eNOS expression and activity and slows the development of atherosclerosis. Activation of GLP-1 signaling in arteries leads to a reduction in inflammatory cytokines. In arterial cells, GLP-1 signaling improves wall disorders induced by various factors including hyperglycemia and inflammatory cytokines. From a clinical perspective, a series of large-scale clinical trials have shown that GLP-1RA has a beneficial effect on combating the onset of cardiovascular events (Kaneto et al., 2021; Zhao et al., 2015).

[0018] Another global health-related problem is the loss of muscle mass and muscle strength, which significantly reduces the quality of life of patients with this condition, such as patients suffering from muscle atrophy due to sarcopenia or cachexia. In addition, muscle atrophy deteriorates the metabolic condition, because healthy muscle mass is beneficial to metabolic regulation. There is a continuous need in the food industry to find better nutritional solutions for patients suffering from such muscle mass or muscle strength loss. In particular, for subjects who cannot perform exercise regimens, a nutritional solution that is effective in itself will be needed. This can also effectively improve the anabolic response to nutritional intake. In addition, dietary supplements that optimize the response of muscle to exercise / muscle contraction are very desirable.

[0019] Studies in rodents have shown that GLP-1RA improves muscle atrophy (Gurjar et al., 2020; Hong et al., 2019; Nguyen et al., 2020). GLP-1 infusion has been further shown to overcome anabolic resistance to feeding in elderly human muscle by enhancing postprandial microvascular perfusion (Abdulla et al., 2020). Therefore, enhanced muscle microvascular perfusion may be a mediator of the ameliorative effects of GLP-1 on muscle atrophy.

[0020] Regarding muscle microvascular perfusion, GLP-1 can increase cardiac and skeletal muscle microvascular blood volume, improve vascular function of conduit vessels, and enhance the metabolic effects of insulin ( Love et al., 2020 ; Subaran et al., 2014 ).

[0021] Muscle microvascular surface area determines substrate and hormone exchange between plasma and muscle interstitium. GLP-1 regulates glucose-dependent insulin secretion and has many extrapancreatic actions, including beneficial vascular effects (Subaran et al., 2014). In a human trial, it was concluded that acute GLP-1 infusion significantly recruited skeletal muscle microvessels and myocardial microvessels in addition to relaxation of conduit arteries in healthy humans. This may contribute to increased tissue oxygen, nutrient and insulin delivery and exchange, and thus better human mealtime glycemic control and tissue function (Subaran et al., 2014).

[0022] It is an object of the present invention to improve upon the prior art and to provide a nutritional solution that addresses the needs expressed above. As disclosed herein, this is facilitated by increasing GLP-1 concentrations in a human subject in need thereof. SUMMARY OF THE INVENTION

[0024] In a first aspect, the present invention provides an agglomerate of soluble whey protein aggregates (AWA) or a dosage form containing an effective amount of AWA for use as a medicament.

[0025] In a further aspect, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject.

[0026] In a further aspect, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the AWA promotes one or more of the following in a human subject:

[0027] i. Increased satiety and / or reduced hunger

[0028] ii. Weight loss

[0029] iii. Improve blood sugar control

[0030] iv. Decreased glucagon secretion

[0031] v. Enhanced insulin secretion

[0032] vi. Decreased body and / or liver fat

[0033] vii. Improve muscle atrophy

[0034] viii. Reduce the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

[0035] In a further aspect, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in treating and / or preventing a condition selected from one or more of the following:

[0036] a. Type II diabetes

[0037] b. Prediabetes

[0038] c. Hyperglycemia

[0039] d. Non-alcoholic fatty liver disease (NAFLD)

[0040] e. Nonalcoholic steatohepatitis (NASH)

[0041] f. Overweight

[0042] g. Obesity

[0043] h. Obesity-related diseases

[0044] i. Muscle atrophy

[0045] j. Cardiovascular disease

[0046] k. Arteriosclerosis

[0047] 1. Hypertension

[0048] m.Dyslipidemia.

[0049] A second aspect of the invention provides a dosage form comprising an effective amount of AWA. The dosage form may be in the form of a nutritional powder comprising at least 30% w / w AWA relative to the amount of total protein, preferably wherein the nutritional powder further comprises at least one non-dairy component, for example in the form of a non-dairy protein or other component not present in milk or whey.

[0050] A third aspect of the present invention provides a method for producing a composition comprising AWA, the method comprising the following steps:

[0051] a. Providing a whey protein solution, the whey protein solution:

[0052] - has a pH in the range of 6-9,

[0053] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0054] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0055] - Contains at least 30% BLG relative to total protein

[0056] b. subjecting the whey protein solution to a heat treatment to obtain a suspension containing sWPA, the heat treatment involving heating the whey protein solution to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of BLG, and

[0057] c. subjecting the suspension or a protein concentrate thereof to a process resulting in agglomeration of the sWPA, preferably freeze drying, and preferably subsequently reducing the particle size of the freeze dried product, thereby obtaining a composition comprising agglomerates of soluble whey protein aggregates.

[0058] A fourth aspect of the present invention provides AWA or a dosage form containing an effective amount of AWA, or a nutritional powder comprising AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA is obtainable by, and for example, by:

[0059] a. Providing a whey protein solution, the whey protein solution:

[0060] -Having a pH in the range of 6-9

[0061] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0062] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0063] - Contains at least 30% BLG relative to total protein

[0064] b. subjecting the whey protein solution to a heat treatment to obtain a suspension containing sWPA, the heat treatment involving heating the whey protein solution to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG, and

[0065] c. subjecting the suspension or a protein concentrate thereof to a process which results in agglomeration of the sWPA, thereby obtaining agglomerates of soluble whey protein aggregates.

[0066] Further aspects of the invention will be apparent from the description of the invention.

[0067] Description of the invention Description of the drawings:

[0069] Figure 1A : Incretin GLP-1 concentrations in young healthy individuals in the post-absorptive state and 6 hours after bolus ingestion of four protein ingredients: MCI, SPI, WPH, and AWA. MCI = Micellar casein isolate; SPI = Serum protein isolate (whey part from milk fractionation); WPH = hydrolyzed SPI; AWA = Agglomerates of soluble whey protein aggregates. Statistical differences (P<0.05) between ingredients within the young group were: AWA was significantly different from ■MCI and ▲WPH, and trended strongly towards a difference relative to ●SPI.

[0070] Figure 1B : Incretin GLP-1 concentrations in healthy elderly individuals in the postabsorptive state and 6 hours after bolus ingestion of four protein ingredients: MCI, SPI, WPH, and AWA. MCI = micellar casein isolate; SPI = whey protein isolate (whey fraction from milk fractionation); WPH = hydrolyzed SPI; AWA = agglomerates of soluble whey protein aggregates. Statistical differences (P < 0.05) between ingredients within the elderly group were: AWA was significantly different from ■MCI, ●SPI, and ▲WPH.

[0071] Figure 2 : Protein (%) of total protein in the supernatant after centrifugation at 3000 x g for 5 min at pH 7.1, 4.2, and 2.9 for sWPA, AWA-f, and AWA-C5b samples.

[0072] Abbreviations, Terms and Definitions:

[0073] In the context of the present invention, the term "soluble whey protein aggregates" (sWPA) refers to submicron-sized aggregates of denatured whey proteins, which typically have a linear, worm-like, branched or chain-like shape. Soluble whey protein aggregates can form strong gels (much stronger than native whey proteins or particulate whey proteins) when present in sufficient concentrations during acidification to pH 4.6. Soluble whey protein aggregates are well known to those skilled in the art and are described, for example, in WO2007 / 110421A2, where they are referred to as linear aggregates, in WO06034856, where they are referred to as activated globular protein preparations, in WO2018 / 011392A1, where they are referred to as acid-gelled whey protein aggregates, in WO2021136785A1, or in US2008305235A1. The amount of soluble whey protein aggregates was quantified according to analysis 3 of WO 2021 / 136785A1. Aqueous solutions of soluble whey protein aggregates are generally transparent, and therefore soluble whey protein aggregates are very suitable for clear beverages.

[0074] In the context of the present invention, the term "agglomerates of soluble whey protein aggregates" or "AWA" relates to sWPA that has been treated to induce secondary aggregation between sWPA and thereby induce agglomeration of sWPA. The inventors have found that aggregation is preferably induced by a treatment that brings the sWPA closer to each other to promote their interaction, for example by removing the solvent (e.g. by freeze drying or concentration of sWPA solutions) and / or by reducing electrostatic repulsion between sWPA.

[0075] The formation of the AWA preferably involves one or more of the following:

[0076] - freeze drying the suspension containing sWPA,

[0077] - incubating the suspension containing sWPA at high ionic strength, for example by increasing the level of divalent metal cations of the sWPA solution having low ionic strength and / or increasing the level of monovalent metal cations,

[0078] -Lowering the pH of the solution containing sWPA

[0079] - concentrating the solution containing sWPA, preferably by one or more of reverse osmosis, nanofiltration, ultrafiltration, microfiltration, evaporation and combinations thereof.

[0080] An example of the production of AWA is found, for example, in US5217741, which is incorporated herein by reference for all purposes.

[0081] In the context of the present invention, the term "β-lactoglobulin" or "BLG" relates to β-lactoglobulin of mammalian species, i.e. in the native, unfolded and optionally also glycosylated form, and includes naturally occurring genetic variants. The term also covers BLG prepared by fermentation. The term does not include denatured BLG. The content of BLG is determined according to assay 6 of WO 2021 / 136785A1.

[0082] In the context of the present invention, the term "α-lactalbumin" or "ALA" relates to α-lactalbumin of mammalian species, i.e. in the native, unfolded and optionally also glycosylated form, and includes naturally occurring genetic variants. The term also covers ALA prepared by fermentation. The term does not include denatured, aggregated ALA. The content of ALA is determined according to assay 6 of WO 2021 / 136785A1.

[0083] In the context of the present invention, the term "type 2 diabetes" or "T2D", formerly known as adult-onset diabetes, is a form of diabetes characterized by hyperglycemia, insulin resistance, and a relative lack of insulin. Long-term complications from hyperglycemia include heart disease, stroke, diabetic retinopathy that may lead to blindness, renal failure, and poor blood flow in limbs that may lead to amputation. Type 2 diabetes occurs primarily due to obesity and lack of exercise. Two cardinal features of T2DM are pancreatic β cell dysfunction and insulin resistance in various insulin target tissues such as liver, skeletal muscle, and adipose tissue. In healthy non-diabetic subjects, β cells can compensate for insulin resistance by increasing insulin secretion or β cell mass, but insufficient compensation leads to the onset of T2DM. After this, once hyperglycemia becomes apparent, β cell function gradually deteriorates, and insulin resistance worsens.

[0084] In the context of the present invention, the term "prediabetes" is a component of the metabolic syndrome and is characterized by elevated blood sugar levels falling below the threshold for diagnosing diabetes. It does not usually cause symptoms, but people with prediabetes often suffer from obesity (especially abdominal or visceral obesity), dyslipidemia with high triglycerides and / or low HDL cholesterol, and hypertension. It is also associated with an increased risk of cardiovascular disease (CVD). More specifically, prediabetes is considered to be an early stage of diabetes because health complications associated with type 2 diabetes often occur before the diagnosis of diabetes.

[0085] In the context of the present invention, the term "hyperglycemia" is the medical term for high blood sugar (glucose) levels. It is a common problem for people with diabetes. It can affect people with type 1 diabetes and type 2 diabetes, as well as pregnant women with gestational diabetes. It may occasionally affect people who do not have diabetes, but usually only affects seriously ill people, such as those who have recently had a stroke or heart attack or have a serious infection. Hyperglycemia should not be confused with hypoglycemia, which is a condition in which a person's blood sugar level drops too low.

[0086] In the context of this invention, the term "non-alcoholic fatty liver disease" or "NAFLD" defines a range of conditions caused by the accumulation of fat in the liver. It is often seen in people who are overweight or obese. Early NAFLD usually does not cause any harm, but if it worsens, it can lead to serious liver damage, including cirrhosis. Having high levels of fat in your liver is also associated with an increased risk of serious health problems, such as diabetes, high blood pressure, and kidney disease. If you already have diabetes, NAFLD increases your risk of developing heart problems. NAFLD is the hepatic manifestation of metabolic syndrome and is characterized by excessive liver fat accumulation associated with insulin resistance.

[0087] In the context of this invention, the term "non-alcoholic steatohepatitis" or "NASH" is liver inflammation and damage caused by a buildup of fat in the liver. It is part of a group of conditions called non-alcoholic fatty liver disease. Many people have a buildup of fat in the liver, and for most people, it causes no symptoms or problems. But in some people, the fat causes inflammation and damages cells in the liver. Because of the damage, the liver does not work as well as it should. NASH can worsen and cause scarring of the liver, which leads to cirrhosis.

[0088] In the context of the present invention, the term "obesity" is defined as a condition in which excess body fat has accumulated to an extent that may have a negative impact on health. Obesity ... 2 People are classified as obese when 2 -30 kg / m 2 range is defined as overweight.

[0089] Obesity is a major cause of disability and is associated with a variety of diseases and conditions. In the context of the present invention, the term "obesity-related disorders" refers to such associated diseases and disorders, such as cardiovascular disease, type 2 diabetes, obstructive sleep apnea, certain types of cancer and osteoarthritis.

[0090] In the context of the present invention, the term "muscle atrophy" refers to the loss of skeletal muscle mass. It may be caused by immobility, aging, malnutrition, medications, or extensive injuries or diseases affecting the musculoskeletal or nervous systems. Muscle atrophy also covers the terms sarcopenia and cachexia. Muscle atrophy results from an imbalance between protein synthesis and protein degradation. Muscle atrophy leads to muscle weakness and causes disability.

[0091] In the context of the present invention, the term "cardiovascular disease" or "CVD" is used as a general term for conditions that affect the heart or blood vessels. This is often associated with a buildup of fatty deposits within the arteries (atherosclerosis) and an increased risk of blood clots. It may also be associated with damage to arteries in organs such as the brain, heart, kidneys and eyes.

[0092] In the context of this invention, the term "atherosclerosis" is used in reference to a condition that occurs when the blood vessels (arteries) that carry oxygen and nutrients from the heart to the rest of the body become thick and stiff - sometimes restricting blood flow to organs and tissues. Healthy arteries are flexible and elastic, but over time, the walls in the arteries can become stiff, a condition often referred to as hardening of the arteries. Atherosclerosis is a specific type of arteriosclerosis. Atherosclerosis is the buildup of fat, cholesterol, and other substances in and on the artery walls. This buildup is called plaque. Plaque can cause the arteries to narrow, blocking blood flow. Plaques can also rupture, causing a blood clot.

[0093] In the context of this invention, the term "hypertension" refers to high blood pressure. If not treated, it increases the risk of serious problems such as heart attack and stroke. Blood pressure is recorded as two numbers. Systolic pressure (the higher number) is the force of the heart pumping blood throughout your body. Diastolic pressure (the lower number) is the resistance to blood flow in the blood vessels. They are both measured in millimeters of mercury (mmHg). As a general guide: High blood pressure is considered to be 140 / 90mmHg or higher (or 150 / 90mmHg or higher if you are over 80 years old).

[0094] In the context of the present invention, the term "dyslipidemia" refers to the abnormal amount of lipids (e.g., triglycerides, cholesterol, and / or adipose phospholipids) in the blood. Dyslipidemia is a risk factor for the development of atherosclerotic cardiovascular disease (ASCVD), such as coronary artery disease, cerebrovascular disease, and peripheral arterial disease. In developed countries, most dyslipidemia is hyperlipidemia; that is, an increase in lipids in the blood. This is usually due to a poor diet and lifestyle. Long-term increases in insulin resistance can also lead to dyslipidemia.

[0095] In the context of the present invention, the term "regular meals" refers to standard meals consumed by subjects as part of their regular daily diet, such as breakfast, lunch and dinner.

[0096] In the context of the present invention, the term "pre-meal" refers to a meal consumed before a regular meal, such as between 1 minute and 180 minutes before consumption of a regular meal.

[0097] The term "lean body mass" (LBM) or "lean body weight" is a body component defined as the difference between total body weight and body fat weight. This means that it counts the mass of all organs, including bones, muscles, blood, skin and everything else, excluding body fat. Lean body mass is determined by dual-energy X-ray absorptiometry (DXA), which is a widely available instrument. Detailed description of the invention:

[0099] The present invention relates to agglomerates of soluble whey protein aggregates (AWA) for use in treating and / or preventing conditions associated with GLP-1 blood levels in a patient.

[0100] I. Agglomerates of soluble whey protein aggregates (AWA)

[0101] Agglomerates of soluble whey protein aggregates (AWA) are a special type of denatured whey protein, which are prepared by denaturing native whey proteins, and in particular β-lactoglobulin, under at least partially demineralized conditions and a suitable pH to form soluble whey protein aggregates (sWPA), and subsequently agglomerating the sWPA to form AWA.

[0102] II. Method for preparing AWA

[0103] In a preferred embodiment of the present invention, the method for producing a composition comprising AWA comprises the following steps:

[0104] a) providing a whey protein solution, said whey protein solution:

[0105] -Having a pH in the range of 6-9

[0106] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0107] - containing 1%-20% w / w BLG relative to the weight of the whey protein solution

[0108] - Contains at least 30% BLG relative to total protein

[0109] b) subjecting the whey protein solution to a heat treatment to obtain a suspension containing sWPA, said heat treatment involving heating the whey protein solution to a temperature in the range of 68°C to 180°C for a period of time sufficient to denature at least 25% w / w of the BLG, and

[0110] c) subjecting the suspension or its protein concentrate to a process resulting in agglomeration of the sWPA, preferably freeze drying, and preferably subsequently reducing the particle size of the freeze dried product,

[0111] Thereby a composition comprising agglomerates of soluble whey protein aggregates is obtained.

[0112] II.i step a)

[0113] The whey protein solution of step a) preferably has a pH in the range of 6-9, more preferably 6.0-9.0, even more preferably 6.3-8.7, and most preferably 6.6-8.5.

[0114] The whey protein solution of step a) preferably has a weight ratio between total protein and the total content of calcium and magnesium of at least 100, more preferably at least 150, even more preferably at least 180, and most preferably at least 190.

[0115] In a preferred embodiment of the present invention, the whey protein solution of step a) has a weight ratio between total protein and the total content of calcium and magnesium in the range of 100-1000, more preferably in the range of 150-1000, even more preferably in the range of 180-1000, and most preferably in the range of 190-1000.

[0116] The weight ratio between total protein and the total content of calcium and magnesium was calculated as the weight of total protein divided by the sum of the weights of calcium and magnesium.

[0117] The whey protein solution of step a) preferably comprises at least 1%-10% w / w BLG relative to the weight of the whey protein solution, more preferably at least 2%-8% w / w relative to the weight of the whey protein solution, even more preferably 2%-6% w / w relative to the weight of the whey protein solution, and most preferably 2%-5% w / w BLG relative to the weight of the whey protein solution.

[0118] The whey protein solution of step a) preferably comprises at least 30% w / w BLG relative to total protein, more preferably at least 40% w / w relative to total protein, even more preferably at least 45% w / w relative to total protein, most preferably at least 50% w / w BLG relative to total protein.

[0119] The whey protein solution is preferably an aqueous solution, and water generally constitutes the majority of the portion of the whey protein solution not made up of solids.

[0120] However, the whey protein solution may contain carbohydrates, preferably limited amounts of carbohydrates. Preferably, the whey protein solution comprises at most 15% w / w carbohydrates relative to the weight of the whey protein solution, more preferably at most 10% w / w relative to the weight of the whey protein solution, even more preferably at most 5% w / w relative to the weight of the whey protein solution, and most preferably at most 1% w / w carbohydrates relative to the weight of the whey protein solution.

[0121] Additionally, the whey protein solution may contain lipids, but preferably only limited amounts of lipids. Preferably, the whey protein solution comprises at most 6% w / w lipids relative to the weight of the whey protein solution, more preferably at most 2% w / w relative to the weight of the whey protein solution, even more preferably at most 0.5% w / w relative to the weight of the whey protein solution, and most preferably at most 0.1% w / w lipids relative to the weight of the whey protein solution.

[0122] The whey protein solution typically has a total solids content in the range of 1%-40% w / w relative to the weight of the whey protein solution, more preferably 2%-30% w / w relative to the weight of the whey protein solution, even more preferably 3%-20% w / w relative to the weight of the whey protein solution and most preferably 3%-10% w / w relative to the weight of the whey protein solution.

[0123] The whey protein of the whey protein solution is preferably provided by one or more of WPC, WPI, SPC and / or SPI.

[0124] In the context of the present invention, the terms "whey protein concentrate" (WPC) and "whey protein concentrate" (SPC) relate to a dry or aqueous composition containing a total amount of protein of 20%-89% w / w relative to total solids.

[0125] The WPC or SPC preferably comprises:

[0126] 20%-89% w / w protein relative to total solids,

[0127] Relative to 15%-70% w / w of total protein in BLG,

[0128] 8%-50% w / w ALA relative to total protein, and

[0129] 0% - 40% w / w casein macropeptide (CMP) relative to protein.

[0130] Optionally, but preferably, the WPC or SPC may contain:

[0131] 20%-89% w / w protein relative to total solids,

[0132] Relative to 15%-90% w / w of total protein in BLG,

[0133] 4%-50% w / w ALA relative to total protein, and

[0134] 0% - 40% w / w CMP relative to protein.

[0135] Preferably, the WPC or SPC comprises:

[0136] 20%-89% w / w protein relative to total solids,

[0137] Relative to 15%-80% w / w of total protein in BLG,

[0138] 4%-50% w / w ALA relative to total protein, and

[0139] 0% - 40% w / w CMP relative to protein.

[0140] More preferably, the WPC or SPC comprises:

[0141] 70%-89% w / w protein relative to total solids,

[0142] Relative to 30%-90% w / w of total protein in BLG,

[0143] 4%-35% w / w ALA relative to total protein, and

[0144] 0% - 25% w / w CMP relative to protein.

[0145] SPC usually contains no CMP or only trace amounts of CMP.

[0146] The terms "whey protein isolate" (WPI) and "whey protein isolate" (SPI) relate to dry or aqueous compositions containing a total amount of protein of 90%-100% w / w relative to total solids.

[0147] The WPI or SPI preferably comprises:

[0148] 90%-100% w / w protein relative to total solids,

[0149] Relative to 15%-70% w / w of total protein in BLG,

[0150] 8%-50% w / w ALA relative to total protein, and

[0151] 0% - 40% w / w CMP relative to total protein.

[0152] Optionally, but preferably, the WPI or SPI may include:

[0153] 90%-100% w / w protein relative to total solids,

[0154] Relative to 30%-95% w / w of total protein in BLG,

[0155] 4%-35% w / w ALA relative to total protein, and

[0156] 0% - 25% w / w CMP relative to total protein.

[0157] More preferably, the WPI or SPI may comprise:

[0158] 90%-100% w / w protein relative to total solids,

[0159] Relative to 30%-90% w / w of total protein in BLG,

[0160] 4%-35% w / w ALA relative to total protein, and

[0161] 0% - 25% w / w CMP relative to total protein.

[0162] SPI usually contains no CMP or only trace amounts of CMP.

[0163] The whey protein used in the whey protein solution is preferably whey protein from mammalian milk, such as, for example, milk from cows, goats, mares, sheep, camels and / or buffaloes. Bovine whey protein is particularly preferred.

[0164] II.ii Step b)

[0165] In step b), a suspension comprising sWPA is obtained by heat treating a demineralized whey protein solution to a temperature of at least 68°C for a period of up to 2 hours.

[0166] Preferably, the demineralized whey protein solution is heated to a temperature in the range of 68-160°C, more preferably 70-145°C, even more preferably 72-120°C, even more preferably 73-100°C, and most preferably 75-95°C.

[0167] In some preferred embodiments of the present invention, the duration of the heat treatment in step b) is sufficient to denature at least 30% w / w of the BLG of the whey protein solution, more preferably at least 50% w / w, even more preferably at least 70% w / w and more preferably at least 90% w / w of the BLG of the whey protein solution.

[0168] The duration of the heat treatment of step b) is preferably in the range of 0.1 s-120 min, more preferably 1 s-60 min, even more preferably 10 s-40 min, even more preferably 0.5 min-40 min, and most preferably 2 min-30 min.

[0169] It is further preferred that step b) comprises a cooling step which reduces the temperature of the obtained sWPA suspension to at most 60°C, more preferably at most 50°C, even more preferably at most 20°C and most preferably at most 15°C.

[0170] The inventors have found that it is advantageous for step b) to provide sWPA in an amount of at most 10% w / w, more preferably at most 8% w / w and most preferably at most 6% w / w relative to the weight of the suspension.

[0171] Preferably, step b) provides sWPA in an amount of 0.5-10% w / w, more preferably 2-8% w / w, and most preferably 3-6% w / w relative to the weight of the suspension.

[0172] In embodiments where the whey protein solution of step a) has a content of BLG and ALA equal to or below the desired content of sWPA, it is generally preferred to denature as much BLG as possible.

[0173] However, in embodiments where the content of BLG and ALA of the whey protein solution of step a) exceeds the desired content of sWPA, it is generally preferred to control the level of denaturation and therefore the level of sWPA formation. This is preferably accomplished by controlling the duration of the heat treatment of step c). The appropriate duration is readily determined by subjecting a sample of the whey protein solution to heat treatment at a given temperature but at three different durations, quantifying the content of sWPA and plotting the resulting concentration of sWPA formed as a function of duration. The duration required for the desired concentration of sWPA can then be determined from the graph.

[0174] Preferably, the heat treatment of step b) has a duration sufficient to provide an amount of sWPA of 0.5-10% w / w, more preferably 2-8% w / w and most preferably 3-6% w / w relative to the weight of the suspension.

[0175] II.iii Step c)

[0176] Step c) of the process involves subjecting the sWPA obtained from step b) to conditions that result in agglomeration of more than one sWPA.

[0177] While this can be achieved in several ways, a currently preferred method involves subjecting a suspension of sWPA to freeze drying, and converting the resulting freeze dried product to a powder, preferably by grinding, milling or other kind of mechanical shearing.

[0178] The sWPA-containing suspension subjected to freeze drying preferably has a pH in the range of 6-9, more preferably 6.0-9.0, even more preferably 6.3-8.7, and most preferably 6.6-8.5.

[0179] Freeze drying preferably involves freezing the suspension containing sWPA to a temperature of at most -5°C, more preferably at most -10°C, and most preferably at most -15°C.

[0180] Freeze drying preferably involves subjecting the frozen suspension to a vacuum to accelerate sublimation of water molecules, preferably at a pressure of at most 500 mBar, more preferably at most 200 mBar, even more preferably at most 50 mBar and more preferably at most 5 mBar.

[0181] The freeze-drying process is preferably stopped when the freeze-dried product contains at most 10% w / w water, more preferably at most 8% water, even more preferably at most 6% water, and most preferably at most 5% water.

[0182] Additionally, it is generally preferred to convert the product obtained from freeze drying directly into a powder by grinding, milling or other kind of mechanical shearing.

[0183] Alternatively, agglomeration can be obtained by concentrating the sWPA obtained from step b) and storing the concentrated suspension for a duration sufficient to form agglomerates. The addition of divalent metal ions, in particular calcium or magnesium, to the suspension or its protein concentrate can initiate the formation of AWA. The AWA-containing suspension can be used as such or undergo further processing, such as heat treatment, optionally further concentration and spray drying.

[0184] In some preferred embodiments of the present invention, the processing leading to agglomeration of the sWPA of step c) involves the addition of divalent metal ions, preferably calcium and / or magnesium.

[0185] Although several sources of divalent metal ions may be used, it is generally preferred that the added divalent metal ions be provided by water-soluble salts of calcium and / or magnesium, preferably CaCl2, MgCl2, CaSO4, MgSO4, CaCO3, MgCO3, calcium phosphate and / or magnesium phosphate.

[0186] The divalent metal ions may be added as solid salts or in dissolved form, usually in water.

[0187] In some preferred embodiments of the invention, the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of at least 1:1, more preferably at least 2:1, even more preferably at least 3:1, and most preferably at least 4:1.

[0188] The total amount of divalent metal ions refers to the total amount in the divalent metal ion-rich liquid after the addition of the metal ions.

[0189] In other preferred embodiments of the present invention, the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of 1:1-100:1, more preferably 2:1-60:1, even more preferably 3:1-40:1, and more preferably 3:1-30:1.

[0190] In a further preferred embodiment of the present invention, the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of 1:1-20:1, more preferably 2:1-16:1, even more preferably 3:1-12:1, and more preferably 3:1-10:1.

[0191] Calcium ions are particularly preferred. In some preferred embodiments of the present invention, the divalent metal ion-rich liquid has a molar percentage of calcium ions of at least 50%, more preferably at least 70%, even more preferably at least 80%, and most preferably at least 90%, relative to the total molar content of divalent metal ions.

[0192] The divalent metal ion-rich liquid is a liquid obtained by adding divalent metal ions and optionally also by moderate pH adjustment.

[0193] Magnesium ions are typically the second most predominant type of divalent metal ion in a divalent metal ion-rich liquid.

[0194] The compositional characteristics and preferences described in the context of the whey protein solution of step a) apply equally to the liquid rich in divalent metal ions, except for the characteristics related to the content of divalent metal ions, and with the exception that a significant portion of the proteins of the whey protein solution is present in the liquid rich in divalent metal ions in denatured form.

[0195] Preferably, the divalent metal ion-rich liquid has a pH in the range of 6-9, more preferably 6.0-9.0, even more preferably 6.3-8.7, and most preferably 6.6-8.5.

[0196] The divalent metal ion-rich liquid is preferably mixed after the addition of the source of divalent metal ions to obtain a uniform distribution of the divalent metal ions.

[0197] The divalent metal ion-rich liquid is then subjected to an incubation step during which the sWPA agglomerates to form AWA.

[0198] In some preferred embodiments of the present invention, the liquid rich in divalent metal ions has a temperature in the range of 10-100°C, more preferably 20-90°C, even more preferably 35-85°C, and most preferably 40-80°C during incubation.

[0199] For example, it may be preferred that the liquid rich in divalent metal ions has a temperature in the range of 10-90°C, more preferably 15-85°C, even more preferably 20-85°C, and most preferably 20-80°C during incubation.

[0200] In some preferred embodiments of the present invention, the divalent metal ion-rich liquid is incubated for at least 1 minute, more preferably at least 2 minutes, and most preferably at least 3 minutes.

[0201] In other preferred embodiments of the present invention, the liquid rich in divalent metal ions is incubated for at least 0.5 hours, more preferably at least 1 hour, and most preferably at least 2 hours.

[0202] In some preferred embodiments of the present invention, the divalent metal ion-rich liquid is incubated for 1 minute to 48 hours, more preferably 2 minutes to 36 hours, and most preferably 3 minutes to 24 hours.

[0203] It is particularly preferred that the liquid rich in divalent metal ions is incubated for 0.5 hours to 48 hours, more preferably for 1 hour to 36 hours, and most preferably for 2 hours to 24 hours.

[0204] It is generally preferred that the liquid rich in divalent metal ions is agitated during the incubation, for example by stirring.

[0205] In some preferred embodiments of the present invention, step c) further involves subjecting the incubated divalent metal ion-rich liquid to one or more of the following:

[0206] o a concentration step, preferably one or more of evaporation, reverse osmosis, nanofiltration, ultrafiltration and / or microfiltration, and

[0207] o A drying step, preferably involving spray drying.

[0208] The drying step involves preheating the liquid to be dried to a temperature in the range of 50°C-80°C before the actual spraying which converts the liquid to be dried into droplets from which the water evaporates.

[0209] In some preferred embodiments of the present invention, the liquid to be dried comprises a warmed liquid rich in divalent metal ions. Preferably, the warmed liquid rich in divalent metal ions accounts for at least 50% w / w of the liquid to be dried, more preferably at least 70% w / w, even more preferably at least 80% w / w and most preferably at least 90% w / w.

[0210] It is generally preferred that the liquid to be dried comprises an incubated divalent metal ion-rich liquid.

[0211] In some preferred embodiments of the invention, the liquid to be dried has a weight percentage of AWA of 2%-10% w / w, more preferably 3%-9% w / w, and most preferably 4%-8% w / w relative to the weight of the liquid.

[0212] It is generally preferred that the liquid to be dried has a weight percentage of AWA relative to total protein of at least 40% w / w, more preferably at least 50% w / w, even more preferably at least 60% w / w and most preferably at least 70% w / w.

[0213] It is particularly preferred that the composition comprising AWA is prepared by a process comprising the steps of

[0214] a) providing a whey protein solution, said whey protein solution:

[0215] -Having a pH in the range of 6.6-8.5

[0216] - has a weight ratio between total protein and total calcium and magnesium content of at least 180

[0217] - comprising at least 2%-5% w / w BLG relative to the weight of the whey protein solution

[0218] - comprises at least 40% w / w BLG relative to total protein,

[0219] b) subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 75°C to 95°C for a period of time sufficient to denature at least 50% of the BLG to obtain a suspension containing sWPA, and

[0220] c) subjecting the suspension or its protein concentrate to freeze drying to prepare a freeze-dried composition, and then subjecting the freeze-dried product to mechanical shearing,

[0221] Thereby a composition comprising agglomerates of soluble whey protein aggregates is obtained.

[0222] It is therefore particularly preferred that the method of the present invention comprises the following steps

[0223] a) providing a whey protein solution, said whey protein solution:

[0224] -Having a pH in the range of 6.6-8.5

[0225] - has a weight ratio between total protein and total calcium and magnesium content of at least 180

[0226] - comprising at least 2%-5% w / w BLG relative to the weight of the whey protein solution

[0227] - comprises at least 40% w / w BLG relative to total protein,

[0228] b) subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 75°C to 95°C for a period of time sufficient to denature at least 50% of the BLG to obtain a suspension containing sWPA, and

[0229] c) subjecting the suspension or its protein concentrate to freeze drying to prepare a freeze-dried composition, and then subjecting the freeze-dried product to mechanical shearing,

[0230] Thereby a composition comprising agglomerates of soluble whey protein aggregates is obtained.

[0231] In other particularly preferred embodiments of the present invention, the method comprises the following steps

[0232] a) providing a whey protein solution, said whey protein solution:

[0233] -Having a pH in the range of 6.6-8.5

[0234] - has a weight ratio between total protein and total calcium and magnesium content of at least 180

[0235] - comprising at least 2%-5% w / w BLG relative to the weight of the whey protein solution

[0236] - comprises at least 40% w / w BLG relative to total protein,

[0237] b) subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 75°C to 95°C for a period of time sufficient to denature at least 50% of the BLG to obtain a suspension containing sWPA, and

[0238] c) subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, said process involving:

[0239] - adding calcium ions and / or magnesium ions in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of 1:1-20:1, more preferably 2:1-16:1, even more preferably 3:1-12:1, and more preferably 3:1-10:1, so that the obtained liquid enriched in divalent metal ions has a molar percentage of calcium ions of at least 50% relative to the total molar content of divalent metal ions,

[0240] - incubating the liquid rich in divalent metal ions at a temperature of 20°C to 80°C for a period of 0.5 hours to 24 hours,

[0241] - optionally concentrating and / or drying the composition obtained from the incubation,

[0242] Thereby a composition comprising AWA is obtained.

[0243] The inventors have found that the content of AWA provided by step c) can be calculated as the reduction in the content of sWPA caused by step c), for example by measuring the content of sWPA before step c) and after step c). If step b) involves drying a suspension of sWPA, quantification requires reconstitution and hydration of the dried product (e.g. at room temperature for 2 hours) before measuring the content of sWPA.

[0244] For example, in Example 1, about 80% of the total protein of the final freeze-dried SPI product was in the form of AWA.

[0245] The composition comprising AWA obtainable by the present process is preferably the same composition as the whey protein solution of step a) with regard to macronutrients, based on total solids. However, the composition comprising AWA obtainable by the present process preferably contains AWA in an amount of at least 20% w / w, more preferably at least 40% w / w, even more preferably at least 60% w / w and more preferably at least 80% w / w relative to total protein.

[0246] The inventors have discovered that AWA prepared by the methods as described herein produces interesting plasma GLP-1 responses when ingested by human subjects.

[0247] Thus, one aspect of the present invention relates to a composition comprising AWA obtainable according to a process as defined herein, preferably in liquid or powder form.

[0248] III. AWA for use in therapeutic methods

[0249] As demonstrated in the Examples section, AWA provides a surprising GLP-1 response in both young and old human subjects compared to administration of a placebo, and compared to administration of other whey protein compositions such as non-heat treated whey fraction (SPI).

[0250] The inventors have surprisingly found that ingestion of AWA induces an increased and prolonged GLP-1 response compared to ingestion of the protein in its native form as well as other protein fractions - see Example 2. Thus, the AWA structure produces improved health benefits compared to other whey proteins. To the best of the inventors' knowledge, this has never been reported.

[0251] In a preferred aspect, the present invention provides an agglomerate of soluble whey protein aggregates (AWA) or a dosage form containing an effective amount of AWA for use as a medicament.

[0252] In a preferred embodiment, the AWA or a dosage form containing an effective amount of the AWA increases and / or prolongs the blood level of GLP-1 in a human subject. In a preferred embodiment, the AWA or a dosage form containing an effective amount of the AWA provides a delayed secretion profile of GLP-1 in a human subject. In a further preferred embodiment, the AWA or a dosage form containing an effective amount of the AWA (i) provides a delayed secretion profile of GLP-1, and (ii) increases and / or prolongs the blood level of GLP-1 in a human subject.

[0253] In a preferred embodiment, the AWA or a dosage form containing an effective amount of the AWA provides a biphasic secretion profile of GLP-1 in a human subject, wherein a relatively small initial secretion occurs, followed by a later increase and prolonged secretion of GLP-1, as measured by blood levels of GLP-1 in the human subject. In a further preferred embodiment, the AWA or a dosage form containing an effective amount of the AWA (i) provides a biphasic secretion profile of GLP-1, and (ii) increases and / or prolongs blood levels of GLP-1 in a human subject.

[0254] Thus, the inventors have discovered a protein composition that induces a delayed but high GLP-1 response in a subject. While not wishing to be bound by theory, it is speculated that AWA appears to induce delayed gastric emptying or slower digestion than native whey protein. In a preferred embodiment, the protein composition provides a delayed but high GLP-1 response in a subject when consumed as a stand-alone meal. In a preferred embodiment, the protein composition provides a delayed but high GLP-1 response in a subject when consumed before, during or after a regular meal.

[0255] In a preferred aspect, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject.

[0256] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in treating and / or preventing one or more conditions associated with decreased blood levels of GLP-1 in a human subject compared to a healthy human subject.

[0257] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in the treatment and / or prevention of one or more conditions in which increased endogenous GLP-1 secretion may be beneficial.

[0258] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the GLP-1 blood level is reduced compared to a healthy human subject, and wherein the human subject would benefit from increased GLP-1 blood levels. III.i Conditions associated with GLP-1 blood levels

[0259] As demonstrated here, AWA affects the level of GLP-1 in the blood of human subjects. The prior art describes further evidence for the many effects of GLP-1 in humans - as disclosed in the Background section.

[0260] The inventors herein have discovered a nutritional solution for increasing and / or prolonging the GLP-1 response, which is most beneficial in increasing satiety and / or reducing hunger in human subjects, which can lead to reduced food intake and / or weight loss in human subjects and thereby provide treatment / reducing the risk of developing overweight and obesity.

[0261] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA increases satiety in the human subject.

[0262] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA reduces hunger in the human subject.

[0263] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA reduces body weight in the human subject.

[0264] The inventors herein have discovered a nutritional solution for increasing and / or prolonging the GLP-1 response, which is further most beneficial in improving glycemic control and / or enhancing insulin secretion and / or reducing glucagon secretion in human subjects, which can lead to a reduced risk of metabolic disease and / or a reduced development of prediabetes and / or type 2 diabetes and related complications.

[0265] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA improves glycemic control in the human subject.

[0266] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA reduces glucagon secretion in the human subject.

[0267] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA enhances insulin secretion in the human subject.

[0268] The inventors herein have discovered a nutritional solution for increasing and / or prolonging the GLP-1 response, which further optimally reduces body and / or liver fat in human subjects, which can lead to reduced risk of developing and / or improving non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0269] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA reduces body and / or liver fat in the human subject.

[0270] The inventors herein have discovered a nutritional solution for increasing and / or prolonging the GLP-1 response, which further optimally increases muscle microvascular perfusion in human subjects, thereby improving tissue oxygen, nutrient and insulin delivery and exchange, better mealtime glycemic control and tissue function and / or overcoming anabolic resistance to feeding and / or ameliorating muscle atrophy.

[0271] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA increases muscle microvascular perfusion in the human subject.

[0272] In a preferred embodiment, the present invention provides an AWA or a dosage form containing an effective amount of an AWA for treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA improves muscle wasting in the human subject.

[0273] The inventors herein have discovered a nutritional solution for increasing and / or prolonging the GLP-1 response, which further optimally facilitates the prevention / reduction of atherosclerosis, hypertension, dyslipidemia and / or cardiovascular events in human subjects.

[0274] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the AWA prevents / reduces arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events in the human subject.

[0275] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject, wherein the one or more conditions associated with GLP-1 blood levels are preferably selected from one or more of the following:

[0276] - Type II diabetes

[0277] - Prediabetes

[0278] - Hyperglycemia

[0279] - Non-alcoholic fatty liver disease (NAFLD)

[0280] - Non-alcoholic steatohepatitis (NASH)

[0281] -overweight

[0282] - Obesity

[0283] -Obesity-related conditions

[0284] -Muscle atrophy

[0285] - Cardiovascular disease

[0286] -arteriosclerosis

[0287] -hypertension

[0288] -Dyslipidemia.

[0289] In a preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for use in treating and / or preventing overweight, obesity and / or obesity-related disorders.

[0290] In another preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing type II diabetes, prediabetes and / or hyperglycemia.

[0291] In another preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH).

[0292] In another preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing muscle atrophy.

[0293] In another preferred embodiment, the present invention provides AWA or a dosage form containing an effective amount of AWA for treating and / or preventing arteriosclerosis, hypertension, dyslipidemia and cardiovascular events.

[0294] III.ii Patient Group

[0295] The AWA or a dosage form containing an effective amount of the AWA is preferably intended as a diet or dietary supplement for a human subject. The human subject who will consume the AWA or the dosage form containing an effective amount of the AWA is preferably an adult. In a preferred embodiment, the human subject is a young adult. In another preferred embodiment, the human subject is a senior. In a preferred embodiment, the human subject is an elderly adult. The term "adult" is defined as a person who is at least 18 years old. The term "young" adult is defined as an adult between 18 and 25 years old. The term "senior" is defined as an adult who is at least 50 years old. The term "senior" adult is defined as a person who is at least 70 years old.

[0296] In a preferred embodiment, the human subjects or patient groups for the treatment and / or prevention of one or more conditions associated with GLP-1 blood levels are at least 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 93, 94, 95, 96, 97, 98, 99, or 100 years of age, or any age in between.

[0297] The human subject can be female or male. In a preferred embodiment, the human subject is male. In another preferred embodiment, the human subject is female.

[0298] In a preferred aspect, the human subjects or patient groups for prevention of one or more conditions associated with GLP-1 blood levels are healthy individuals, i.e., individuals who do not exhibit typical symptoms of conditions associated with GLP-1 blood levels (such as those listed in Section III.i) and / or have not been diagnosed as having these conditions, as would be recognized by a medical professional.

[0299] In another preferred aspect, human subjects or patient groups for treatment of one or more conditions associated with GLP-1 blood levels are individuals who exhibit typical symptoms of those conditions associated with GLP-1 blood levels (such as those listed in Section III.i), who may or may not have been diagnosed by a medical professional.

[0300] In one such preferred embodiment, the human subject is overweight or obese."Overweight" is defined as having a body mass index of at least 25.0. "Obese" is defined as having a body mass index of at least 30.0.

[0301] In one such preferred embodiment, the human subject has high blood glucose levels, such as characterized by fasting and 2 hour postprandial blood glucose levels greater than 125 mg / dl and 180 mg / dl, respectively.

[0302] In one such preferred embodiment, the human subject has fat accumulation in the liver, such as characterized by steatosis in >5% of hepatocytes or >5% triglycerides in wet liver tissue weight.

[0303] In one such preferred embodiment, the human subject has abnormal amounts of lipids in the blood (dyslipidemia), such as characterized by total cholesterol levels >5mmol / l, HDL (high-density lipoprotein) levels <1mmol / l, triglyceride levels >2mmol / l or LDL (low-density lipoprotein) levels >3mmol / l.

[0304] In one such preferred embodiment, the human subject has an accumulation of fatty deposits in the arteries, such as characterized by hypertension, wherein the systolic blood pressure (the higher number) and diastolic blood pressure (the lower number) are considered to be 140 / 90 mmHg or higher; or 150 / 90 mmHg or higher if the subject is over 80 years of age.

[0305] In one such preferred embodiment, the human subject suffers from loss of skeletal muscle mass / muscle atrophy caused by sarcopenia or cachexia or disuse, such as characterized by low muscle strength, low muscle quantity or quality, or low physical performance, such as specified in Table 3 in Cruz-Jentoft et al. 2019, which provides suggestions for cutoff points for different parameters to increase coordination of sarcopenia studies.

[0306] III.iii Administration and Dosage Regimen

[0307] In a preferred embodiment, the AWAs of the present invention are intended for oral administration to human subjects.

[0308] In a preferred embodiment, an AWA or a dosage form comprising an AWA for use according to the invention is administered to a human subject in an amount sufficient to provide a dose of AWA of at least 0.01 g / kg, 0.02 g / kg, 0.03 g / kg, 0.04 g / kg, 0.05 g / kg, 0.06 g / kg, 0.07 g / kg, 0.08 g / kg, 0.09 g / kg, 0.1 g / kg, 0.2 g / kg, 0.3 g / kg, 0.4 g / kg, 0.5 g / kg, 0.6 g / kg, 0.7 g / kg, 0.8 g / kg, 0.9 g / kg, 1.0 g / kg, 1.1 g / kg, 1.2 g / kg, 1.3 g / kg, 1.4 g / kg, 1.5 g / kg, 1.6 g / kg, 1.7 g / kg, 1.8 g / kg, 1.9 g / kg or 2.0 g / kg of lean body weight per dose.

[0309] In a preferred embodiment, the AWA or dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose.

[0310] In a preferred embodiment, AWA or a dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-4 g / kg lean body weight per dose, preferably 0.05 g / kg-2 g / kg lean body weight per dose, preferably 0.1 g / kg-1 g / kg lean body weight per dose, more preferably 0.2 g / kg-0.5 g / kg lean body weight per dose.

[0311] AWA can be applied once a day or several times a day, such as in the morning, at noon and / or in the evening. In a preferred embodiment, AWA is applied between 1-5 times a day, such as once, twice, three times, four times or five times a day.

[0312] In a preferred embodiment, the AWA for use according to the invention or a dosage form comprising the AWA is in an amount sufficient to provide at least 0.01 g / kg, 0.02 g / kg, 0.03 g / kg, 0.04 g / kg, 0.05 g / kg, 0.06 g / kg, 0.07 g / kg, 0.08 g / kg, 0.09 g / kg, 0.1 g / kg, 0.2 g / kg, 0.3 g / kg, 0.4 g / kg, 0.5 g / kg, 0.6 g / kg, 0.7 g / kg, 0.8 g / kg, 0.9 g / kg, 1.0 g / kg, 1.1 g / kg, 1.2 g / kg, 1.3 g / kg, 1.4 g / kg, 1.5 g / kg, 1.6 g / kg, 1.7 g / kg, 1.8 g / kg, 1.9 g / kg, 2.10 g / kg, 2.2 g / kg, 2.3 g / kg, 2.4 g / kg, 2.5 g / kg, 2.6 g / kg, 2.7 g / kg, or 4.5 g / kg body weight per day of AWA.

[0313] In a preferred embodiment, the AWA or dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dosage of AWA of at least 0.05 g / kg body weight per day, preferably at least 0.1 g / kg body weight per day, more preferably at least 0.3 g / kg body weight per day, even more preferably at least 0.5 g / kg body weight per day.

[0314] In a preferred embodiment, the AWA or dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-4 g / kg body weight per day, preferably 0.05 g / kg-2 g / kg body weight per day, preferably 0.1 g / kg-1 g / kg body weight per day, more preferably 0.2 g / kg-0.5 g / kg body weight per day.

[0315] In preferred embodiments, the AWA or dosage form comprising the AWA for use according to the invention is sufficient to provide at least 0.01 g / kg, 0.02 g / kg, 0.03 g / kg, 0.04 g / kg, 0.05 g / kg, 0.06 g / kg, 0.07 g / kg, 0.08 g / kg, 0.09 g / kg, 0.1 g / kg, 0.2 g / kg, 0.3 g / kg, 0.4 g / kg, 0.5 g / kg, 0.6 g / kg, 0.7 g / kg, 0.8 g / kg, 0.9 g / kg, 1.0 g / kg, 1.1 g / kg, 1.2 g / kg, 1.3 g / kg, 1.4 g / kg, 1.5 g / kg, 1.6 g / kg, 1.7 g / kg, 1.8 g / kg, 1.9 g / kg, 2.1 g / kg, 2.2 g / kg, 2.3 g / kg, 2.4 g / kg, 2.5 g / kg, 2.6 g / kg, 2.7 g / kg, 2.8 g / kg or 4.5 g / kg of AWA per day.

[0316] In a preferred embodiment, the AWA or dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dosage of AWA of at least 0.05 g / kg lean body weight per day, preferably at least 0.1 g / kg lean body weight per day, more preferably at least 0.3 g / kg lean body weight per day, even more preferably at least 0.5 g / kg lean body weight per day.

[0317] In a preferred embodiment, AWA or a dosage form comprising AWA for use according to the present invention is administered to a human subject in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-2 g / kg lean body weight per day, preferably 0.05 g / kg-1 g / kg lean body weight per day, preferably 0.1 g / kg-0.5 g / kg lean body weight per day, more preferably 0.2 g / kg-0.5 g / kg lean body weight per day.

[0318] The AWA or a dosage form containing an effective amount of the AWA is intended for use as a diet or dietary supplement for human subjects.

[0319] In a preferred embodiment, the AWA of the present invention is administered as a separate meal.Using AWA as such a meal may be particularly advantageous in the treatment and / or prevention of overweight, obesity and / or metabolic disorders.

[0320] In a preferred embodiment, the AWA of the present invention is intended to be administered as a pre-meal meal.

[0321] In a preferred embodiment of the invention, the AWA for use according to the invention is administered to a human subject as a pre-meal meal, such as between 5 minutes and 180 minutes before a regular meal, between 10 minutes and 180 minutes before a regular meal, between 15 minutes and 180 minutes before a regular meal, between 20 minutes and 180 minutes before a regular meal, between 25 minutes and 180 minutes before a regular meal, between 30 minutes and 180 minutes before a regular meal, between 35 minutes and 180 minutes before a regular meal, between 40 minutes and 180 minutes before a regular meal, between 45 minutes and 180 minutes before a regular meal, between 50 minutes and 180 minutes before a regular meal, between 55 minutes and 180 minutes before a regular meal, between 60 minutes and 180 minutes before a regular meal, between 75 minutes and 180 minutes before a regular meal, between 80 minutes and 180 minutes before a regular meal, between 85 minutes and 180 minutes before a regular meal, between 90 minutes and 180 minutes before a regular meal. between 95 minutes and 180 minutes before regular meals, between 100 minutes and 180 minutes before regular meals, between 105 minutes and 180 minutes before regular meals, between 110 minutes and 180 minutes before regular meals, between 115 minutes and 180 minutes before regular meals, between 120 minutes and 180 minutes before regular meals, between 125 minutes and 180 minutes before regular meals, between 130 minutes and 180 minutes before regular meals, between 135 minutes and 150 minutes before regular meals The invention may be administered between 120 minutes and 180 minutes before a regular meal, between 140 minutes and 180 minutes before a regular meal, between 145 minutes and 180 minutes before a regular meal, between 150 minutes and 180 minutes before a regular meal, between 155 minutes and 180 minutes before a regular meal, between 160 minutes and 180 minutes before a regular meal, between 165 minutes and 180 minutes before a regular meal, between 170 minutes and 180 minutes before a regular meal, or between 175 minutes and 180 minutes before a regular meal.

[0322] In a preferred embodiment of the invention, the AWA for use according to the invention is administered to a human subject as a pre-meal meal, such as between 5 minutes and 30 minutes before a regular meal, preferably between 30 minutes and 60 minutes before a regular meal, more preferably between 60 minutes and 90 minutes, preferably between 90 minutes and 120 minutes, preferably between 120 minutes and 150 minutes, preferably between 150 minutes and 180 minutes before a regular meal.

[0323] Using AWA as such a pre-meal meal may be particularly advantageous in connection with the treatment and / or prevention of obesity / overweight to reduce hunger resulting in reduced food intake.

[0324] In a preferred embodiment of the invention, the AWA for use according to the invention is administered to a human subject as a pre-meal meal, such as between 1 minute-180 minutes before a regular meal, between 1 minute-175 minutes before a regular meal, between 1 minute-170 minutes before a regular meal, between 1 minute-165 minutes before a regular meal, between 1 minute-160 minutes before a regular meal, between 1 minute-155 minutes before a regular meal, between 1 minute-150 minutes before a regular meal, between 1 minute-145 minutes before a regular meal, between 1 minute-140 minutes before a regular meal, between 1 minute-135 minutes before a regular meal, between 1 minute-130 minutes before a regular meal, between 1 minute-125 minutes before a regular meal, between 1 minute-120 minutes before a regular meal, between 1 minute-115 minutes before a regular meal, between 1 minute-110 minutes before a regular meal, between 1 minute-10 ... 1 minute-100 minutes before a regular meal, between 1 minute-95 minutes before a regular meal, between 1 minute-90 minutes before a regular meal, between 1 minute-85 minutes before a regular meal, between 1 minute-80 minutes before a regular meal, between 1 minute-75 minutes before a regular meal, between 1 minute-70 minutes before a regular meal, between 1 minute-65 minutes before a regular meal, between 1 minute-60 minutes before a regular meal, between 1 minute-55 minutes before a regular meal, between 1 minute-50 minutes before a regular meal, between 1 minute-45 minutes before a regular meal, between 1 minute-40 minutes before a regular meal, between 1 minute-35 minutes before a regular meal, between 1 minute-30 minutes before a regular meal, between 1 minute-25 minutes before a regular meal, between 1 minute-20 minutes before a regular meal, between 1 minute-15 minutes before a regular meal, between 1 minute-10 minutes before a regular meal, or between 1 minute-5 minutes before a regular meal.

[0325] In a preferred embodiment of the present invention, the AWA for use according to the present invention is administered to a human subject as a pre-meal meal, such as between 150 minutes to 180 minutes before a regular meal, between 120 minutes to 150 minutes before a regular meal, between 90 minutes to 120 minutes before a regular meal, between 60 minutes to 90 minutes before a regular meal, preferably between 30 minutes to 60 minutes before a regular meal, more preferably between 15 minutes to 30 minutes, more preferably between 1 minute to 15 minutes.

[0326] The use of AWA as such a pre-meal supplement may be particularly advantageous in the treatment and / or prevention of obesity / overweight to increase satiety leading to reduced food intake.

[0327] In a preferred embodiment, the AWA of the present invention is intended to be administered in combination with a meal, such as ingested simultaneously with a meal.

[0328] In a preferred embodiment of the invention, the AWA for use according to the invention is administered to a human subject in combination with a meal. Thus, the GLP-1 response produced by the meal is additive to the GLP-1 peak produced by the AWA, which peak is delayed, prolonged and increased (relative to the first peak). Thus, the resulting overall GLP-1 response is extended and prolonged in time.

[0329] In another preferred embodiment, the AWA for use according to the invention, such as in particular with regard to the treatment and / or prevention of muscle atrophy, can be administered to a human subject before and / or after surgery and / or hospitalization, so that advantageously the subject has improved his muscle mass before undergoing surgery or a longer bedridden hospitalization, and continues to maintain such elevated GLP-1 concentrations during the recovery period to continue to improve muscle mass. This provides an optimal nutritional state to minimize the loss of muscle mass during the hospital intervention, and also prepares him for a faster recovery and accumulation of lost muscle tissue thereafter.

[0330] The treatment or prevention provided by the present invention benefits from an additional exercise regimen, and the effectiveness of the present treatment or prevention does not require an additional exercise regimen. Thus, in one embodiment, the present invention provides a treatment and / or prevention of one or more GLP-1 related disorders that does not include an additional exercise regimen.

[0331] IV. Dosage Forms Containing AWA

[0332] Those skilled in the art will understand that they can freely combine all features of the invention disclosed herein. In particular, features described for therapeutic use can be used and combined with features of the dosage form of the AWA product, and vice versa.

[0333] Therefore, a further preferred aspect of the present invention relates to a dosage form comprising an effective amount of AWA as defined herein.

[0334] IV.i Types of Dosage Forms

[0335] While all types of dosage forms may be used, it is particularly preferred that the dosage form is selected from the group consisting of liquids, powders, gels, capsules and / or edible bars.

[0336] A particularly preferred dosage form is a "ready-to-drink beverage" (RTD), ie a liquid formulated for drinking as is, preferably pasteurized or even aseptic, and packaged in a sealed container which allows the RTD beverage to be stored at ambient temperature or under refrigeration.

[0337] Another particularly preferred dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0338] When the term "effective amount of an AWA" is referred to herein with respect to a dosage form without reference to a specific disease or condition, it is preferred that the AWA is present in the dosage form in an amount sufficient to provide treatment or prevention of at least one of the diseases and conditions mentioned herein.

[0339] The dosage form preferably has a pH in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5. In another preferred embodiment, the pH of the dosage form is acidic, preferably having a pH in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0340] The pH value mentioned herein refers to the pH value when measured according to assay 13 of WO 2021 / 136785.

[0341] IV.ii Composition of the dosage form

[0342] In some preferred embodiments of the present invention, the dosage form comprises AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w relative to total protein, even more preferably at least 60% w / w relative to total protein, and more preferably at least 70% w / w relative to total protein.

[0343] In some preferred embodiments of the present invention, the dosage form comprises an amount of AWA of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose. As previously disclosed, the AWA can be administered once a day or several times a day. The amount of AWA in the dosage form can be affected by the choice of the number of dose administrations.

[0344] In some preferred embodiments of the present invention, liquid dosage forms such as, for example, RTDs are particularly preferred, the dosage forms comprising AWA in an amount of preferably at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0345] In some preferred embodiments of the present invention, liquid dosage forms are particularly preferred, such as, for example, RTD, the dosage form comprising AWA in an amount of 1 g / 100 mL-20 g / 100 mL, more preferably 2 g / 100 mL-18 g / 100 mL, even more preferably at least 3 g / 100 mL-14 g / 100 mL, and most preferably 5 g / 100 mL-10 g / 100 mL.

[0346] In some preferred embodiments of the present invention, liquid dosage forms such as, for example, RTDs are particularly preferred, the dosage forms comprising total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0347] In some preferred embodiments of the present invention, liquid dosage forms are particularly preferred, such as, for example, RTD, the dosage form comprising total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL, and most preferably 4 g / 100 mL-14 g / 100 mL.

[0348] In some preferred embodiments of the present invention, particularly preferred for powders intended to be dissolved in a liquid prior to ingestion, such as milkshake powders, the dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g, and most preferably at least 50 g / 100 g.

[0349] In some preferred embodiments of the present invention, particularly preferred for powders intended to be dissolved in a liquid prior to ingestion, such as milkshake powders, the dosage form comprises AWA in an amount of at least 50 g / 100 g, more preferably at least 60 g / 100 g, more preferably at least 70 g / 100 g, even more preferably at least 80 g / 100 g, and most preferably at least 90 g / 100 g.

[0350] In some preferred embodiments of the present invention, particularly preferably for powders intended to be dissolved in a liquid prior to ingestion, such as milkshake powders, the dosage form comprises AWA in an amount of 10 g / 100 g - 90 g / 100 g, more preferably 15 g / 100 g - 85 g / 100 g, even more preferably at least 20 g / 100 g - 80 g / 100 g, and most preferably 30 g / 100 g - 75 g / 100 g.

[0351] The dosage form may also contain non-protein macronutrients and micronutrients, preferably as described on pages 62-65 of WO 2021 / 136785, which is incorporated herein by reference.

[0352] In a preferred embodiment, the present invention provides a nutritional powder comprising

[0353] - AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein, and

[0354] - At least one non-dairy ingredient.

[0355] The dosage form may contain other macronutrients besides protein, such as, for example, carbohydrates and / or lipids.

[0356] In some embodiments of the invention, the dosage form further comprises carbohydrates. The total carbohydrate content in the dosage form depends on the intended use of the dosage form.

[0357] The carbohydrates of the dosage form are preferably provided by one or more sources of carbohydrate sources.

[0358] Useful carbohydrate sources can be selected from the group consisting of: sucrose, maltose, dextrose, galactose, maltodextrin, corn syrup solids, sucromalt, glucose polymers, corn syrup, modified starch, resistant starch, carbohydrates derived from rice, isomaltulose, white sugar, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols, fructooligosaccharides, soy fiber, corn fiber, guar gum, konjac flour, polydextrose, fibersol and combinations thereof. In some embodiments of the present invention, the dosage form comprises non-digestible sugars such as fructans, and the fructans comprise inulin or fructooligosaccharides.

[0359] In some preferred embodiments of the invention, the dosage form comprises carbohydrates in a range between 0% and 95% of the total energy content of the dosage form, more preferably in a range between 10% and 85% of the total energy content of the dosage form, even more preferably in a range between 20% and 75% of the total energy content of the dosage form, and most preferably in a range between 30% and 60% of the total energy content of the dosage form.

[0360] Even lower carbohydrate contents are generally preferred, thus in some preferred embodiments of the invention the dosage form comprises carbohydrates in an amount in the range between 0% and 30% of the total energy content of the dosage form, more preferably in the range between 0% and 20% of the total energy content of the dosage form, and even more preferably in the range between 0% and 10% of the total energy content of the dosage form.

[0361] In some preferred embodiments of the invention, the dosage form comprises carbohydrates in a total amount of at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the dosage form.

[0362] In some preferred embodiments of the invention, the dosage form is particularly useful as a nutritional supplement for nutritionally incomplete individuals and comprises a total amount of carbohydrates in the range of between 70%-95%, preferably between 80E%-90E% of the total energy content (E) of the dosage form.

[0363] In some preferred embodiments of the invention, the dosage form comprises carbohydrates in a total amount in the range of between 30%-60% of the total energy content of the dosage form, and most preferably in the range of between 35E%-50E%. Such dosage forms are particularly useful for nutritionally complete dosage forms.

[0364] In some embodiments of the invention, the dosage form further comprises at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins, and combinations thereof.

[0365] Additional ingredients may be used to tailor the nutritional contribution as well as the taste and flavor characteristics of the dosage form.

[0366] In one embodiment of the present invention, the dosage form comprises at least one high intensity sweetener (HIS). At least one HIS is preferably selected from the group consisting of aspartame, cyclamate, sucralose, acesulfame potassium salt, neotame, saccharin, stevia extract, steviol glycosides such as, for example, steviol bioside A, or a combination thereof.

[0367] In some embodiments of the invention, it is particularly preferred that the sweetener comprises or even consists of one or more high intensity sweeteners.

[0368] HIS is present in both natural and artificial sweeteners and generally has a sweetness intensity at least 10 times that of sucrose.

[0369] If used, the total amount of HIS of the dosage form is generally in the range of 0.001%-2% w / w. Preferably, the total amount of HIS is in the range of 0.005%-1% w / w. Most preferably, the total amount of HIS is in the range of 0.01%-0.5% w / w.

[0370] The choice of sweetener may depend on the dosage form to be produced, for example, high intensity sweeteners (e.g., aspartame, acesulfame-K, or sucralose) may be used in dosage forms where the energy contribution from the sweetener is not desired, while for dosage forms with a natural profile, natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used.

[0371] In addition, it may be preferred that sweetener comprises one or more polyol sweeteners or even consists of one or more polyol sweeteners.The limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol or its combination.If used, the total amount of the polyol sweetener of dosage form is usually in the scope of 1%-20%w / w.More preferably, the total amount of the polyol sweetener of dosage form is in the scope of 2%-15%w / w.Even more preferably, the total amount of polyol sweetener can be in the scope of 4%-10%w / w.

[0372] In some preferred embodiments of the present invention, the dosage form comprises:

[0373] - a total amount of at most 1% w / w, more preferably at most 0.5% w / w, and most preferably at most 0.1% w / w of carbohydrates, and

[0374] - A total amount of HIS in the range of 0.001%-2% w / w, more preferably in the range of 0.005%-1% w / w, and most preferably in the range of 0.01%-0.5% w / w.

[0375] In some embodiments of the invention, the dosage form further comprises lipids. The total lipid content in the dosage form of the invention depends on the intended use of the dosage form.

[0376] In some preferred embodiments of the present invention, the dosage form has a lipid content between 0% and 50% of the total energy content of the dosage form, more preferably in the range between 0% and 40% of the total energy content of the dosage form, even more preferably in the range between 0% and 30% of the total energy content of the dosage form, even more preferably in the range between 0% and 20% of the total energy content of the dosage form, even more preferably in the range between 0% and 10% of the total energy content of the dosage form, and most preferably in the range between 0% and 5% of the total energy content of the dosage form.

[0377] In some preferred embodiments of the invention, the dosage form is particularly useful as a sports dosage form, ie a dosage form which can advantageously be taken before, during or after physical exercise, and comprises a total amount of lipids of, for example, at most 10E%, preferably at most 1E%.

[0378] In some preferred embodiments of the invention, the dosage form is particularly useful as a nutritional supplement for nutritionally incomplete persons and comprises a total amount of lipids of at most 10%, preferably at most 1E%, of the total energy content of the dosage form.

[0379] In some preferred embodiments of the present invention, the dosage form, for example in the form of a sports dosage form, comprises:

[0380] - a total amount of proteins in the range of 1%-30% w / w relative to the weight of the dosage form, more preferably 4%-24% w / w relative to the weight of the dosage form, even more preferably 8%-20% w / w relative to the weight of the dosage form, and most preferably 10%-20% w / w relative to the weight of the dosage form,

[0381] - carbohydrates in a total amount of at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the dosage form, and

[0382] - a total amount of at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% lipids.

[0383] In other preferred embodiments of the invention, the dosage form, for example in the form of a low carbohydrate sports dosage form, comprises:

[0384] - a total amount of proteins in the range of 1%-30% w / w relative to the weight of the dosage form, more preferably 4%-24% w / w relative to the weight of the dosage form, even more preferably 8%-20% w / w relative to the weight of the dosage form, and most preferably 10%-20% w / w relative to the weight of the dosage form,

[0385] - a total amount of at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of carbohydrates,

[0386] - a total amount of at most 5E%, more preferably at most 4E%, even more preferably at most 3E%, and most preferably at most 1E% lipids, and

[0387] - A total amount of HIS in the range of 0.001%-2% w / w, more preferably in the range of 0.005%-1% w / w, and most preferably in the range of 0.01%-0.5% w / w.

[0388] In other preferred embodiments of the present invention, the dosage form, for example in the form of a nutritionally complete dosage form, comprises:

[0389] - a total amount of proteins in the range of 1%-30% w / w relative to the weight of the dosage form, more preferably 4%-24% w / w relative to the weight of the dosage form, even more preferably 8%-20% w / w relative to the weight of the dosage form, and most preferably 10%-20% w / w relative to the weight of the dosage form,

[0390] - a total amount of carbohydrates in the range between 30%-60% of the total energy content of the dosage form, and most preferably in the range between 35E%-50E%, and

[0391] - A total amount of lipids in the range of 20%-50%, more preferably in the range of 25E%-45E%, and most preferably 30E%-40E% of the total energy content.

[0392] In some preferred embodiments of the present invention, the AWA used to prepare the dosage form is provided by an AWA-containing composition obtainable by the methods described herein.

[0393] Preferably, the preparation of the dosage form involves mixing the AWA-containing composition obtainable by the process described herein with one or more additional ingredients required for the dosage form.

[0394] The dosage form of the invention can advantageously be used as a pre-meal meal.

[0395] V. Treatment Methods

[0396] In a preferred aspect, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0397] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, and wherein increasing GPL-1 levels is beneficial in treating / preventing the condition.

[0398] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, wherein the AWA increases satiety / reduces hunger in the human subject.

[0399] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering an AWA or a dosage form containing an effective amount of an AWA to a human subject, wherein the AWA enhances insulin secretion, reduces glucagon secretion and / or improves glycemic control in the human subject.

[0400] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, wherein the AWA reduces the body weight of the human subject.

[0401] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, wherein the AWA reduces body and / or liver fat in the human subject.

[0402] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, wherein the AWA improves muscle atrophy in the human subject.

[0403] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject, wherein the AWA reduces arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

[0404] In a preferred embodiment, the present invention provides a method for treating and / or preventing one or more conditions selected from the following list: type II diabetes, prediabetes, hyperglycemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), overweight, obesity, obesity-related conditions, muscle atrophy, cardiovascular disease, arteriosclerosis, hypertension and dyslipidemia; wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0405] In a preferred embodiment, the present invention provides a method for treating and / or preventing overweight, obesity and / or obesity-related disorders; wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0406] In a preferred embodiment, the present invention provides a method for treating and / or preventing type II diabetes, prediabetes and / or hyperglycemia; wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0407] In a preferred embodiment, the present invention provides a method for treating and / or preventing non-alcoholic fatty liver disease (NAFLD) and / or non-alcoholic steatohepatitis (NASH); wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0408] In a preferred embodiment, the present invention provides a method for treating and / or preventing muscle atrophy; wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0409] In a preferred embodiment, the present invention provides a method for treating and / or preventing arteriosclerosis, hypertension, dyslipidemia and cardiovascular events; wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0410] VI. Non-therapeutic treatments

[0411] In a preferred aspect, the present invention provides a method for increasing and / or prolonging glucagon-like peptide 1 (GLP-1) secretion in a human subject, preferably a non-therapeutic method, wherein the method comprises administering to the human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0412] In a preferred embodiment, the present invention provides a method, preferably a non-therapeutic method, for reducing hunger and / or increasing satiety in a human subject, wherein the method comprises administering to the human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0413] In a preferred embodiment, the present invention provides a method, preferably a non-therapeutic method, for reducing body weight in a human subject, wherein the method comprises administering to the human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0414] In a preferred embodiment, the present invention provides a method, preferably a non-therapeutic method, for reducing body and / or liver fat in a human subject, wherein the method comprises administering to the human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0415] In a preferred embodiment, the present invention provides a method, preferably a non-therapeutic method, for enhancing insulin secretion, reducing glucagon secretion and / or improving glycemic control in a human subject, wherein the method comprises administering to the human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0416] In a preferred embodiment, the present invention provides a method for improving muscle atrophy, preferably a non-therapeutic method, wherein the method comprises administering an effective amount of AWA or a dosage form containing an effective amount of AWA to a human subject.

[0417] In a preferred embodiment, the present invention provides a method, preferably a non-therapeutic method, for reducing arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events, wherein the method comprises administering to a human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0418] VII. Uses of AWA

[0419] In a preferred aspect, the present invention relates to the use of an effective amount of AWA or a dosage form containing an effective amount of AWA for increasing and / or prolonging the secretion of glucagon-like peptide 1 (GLP-1) in a human subject.

[0420] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for reducing hunger and / or increasing satiety in a human subject.

[0421] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for reducing body and / or liver fat in a human subject.

[0422] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for reducing body weight in a human subject.

[0423] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for enhancing insulin secretion, reducing glucagon secretion and / or improving glycemic control in a human subject.

[0424] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for improving muscle atrophy in a human subject.

[0425] In a preferred embodiment, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA for reducing arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

[0426] VIII. Use of AWA in the preparation of medicines

[0427] In a preferred aspect, the present invention relates to the use of AWA or a dosage form containing an effective amount of AWA in the preparation of a medicament for treating and / or preventing one or more of the following:

[0428] - Type II diabetes

[0429] - Prediabetes

[0430] - Hyperglycemia

[0431] - Non-alcoholic fatty liver disease (NAFLD)

[0432] - Non-alcoholic steatohepatitis (NASH)

[0433] -overweight

[0434] - Obesity

[0435] -Obesity-related conditions

[0436] -Muscle atrophy

[0437] - Cardiovascular disease

[0438] -arteriosclerosis

[0439] -hypertension

[0440] -Dyslipidemia.

[0441] Preferred numbering implementation scheme

[0442] In the following, preferred numbered embodiments of the present invention are described.

[0443] Preferred numbered embodiments 1. Agglomerates of soluble whey protein aggregates (AWA) or dosage forms containing an effective amount of AWA for use as a medicament.

[0444] Preferred numbered embodiment 2. The AWA according to any of the preceding preferred numbered embodiments or a dosage form containing an effective amount of the AWA for use in treating and / or preventing one or more disorders associated with GLP-1 blood levels in a human subject.

[0445] Preferred numbered embodiment 3. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA increases and / or prolongs the blood level of GLP-1 in a human subject.

[0446] Preferred numbered embodiment 4. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA provides a biphasic secretion profile of GLP-1 in a human subject.

[0447] Preferred numbered embodiment 5. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA provides a delayed secretion profile of GLP-1 in a human subject.

[0448] Preferred numbered embodiment 6. The AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA increases satiety in a human subject.

[0449] Preferred numbered embodiment 7. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA reduces hunger in a human subject.

[0450] Preferred numbered embodiment 8. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA reduces body weight in a human subject.

[0451] Preferred numbered embodiment 9. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA improves glycemic control in a human subject.

[0452] Preferred numbered embodiment 10. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA enhances insulin secretion in a human subject.

[0453] Preferred numbered embodiment 11. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA reduces glucagon secretion in a human subject.

[0454] Preferred numbered embodiment 12. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA reduces body and / or liver fat in a human subject.

[0455] Preferred numbered embodiment 13. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA improves muscle atrophy in a human subject.

[0456] Preferred numbered embodiment 14. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA reduces the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events in a human subject.

[0457] Preferred numbered embodiment 15. AWA for treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the disorder associated with GLP-1 blood levels is selected from one or more of the following:

[0458] a. Type II diabetes

[0459] b. Prediabetes

[0460] c. Hyperglycemia

[0461] d. Non-alcoholic fatty liver disease (NAFLD)

[0462] e. Non-alcoholic steatohepatitis (NASH)

[0463] f. Overweight

[0464] g. Obesity

[0465] h. Obesity-related diseases

[0466] i. Muscle atrophy

[0467] j. Cardiovascular disease

[0468] k. Arteriosclerosis

[0469] 1. Hypertension

[0470] m.Dyslipidemia.

[0471] Preferred numbered embodiment 16. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the human subject is an adult, and more preferably an elderly person.

[0472] Preferred numbered embodiment 17. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the human subject is a young adult.

[0473] Preferred numbered embodiment 18. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any one of the preceding preferred numbered embodiments, wherein the human subject is a healthy individual.

[0474] Preferred numbered embodiment 19. AWA or a dosage form containing an effective amount of AWA for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the human subject is overweight and more preferably obese.

[0475] Preferred numbered embodiment 20. An AWA for use in treatment and / or prevention or a dosage form containing an effective amount of an AWA according to any one of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is for oral administration to a human subject.

[0476] Preferred numbered embodiment 21. An AWA for treatment and / or prevention or a dosage form containing an effective amount of an AWA according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in combination with a meal, preferably as a pre-meal meal, between 5 minutes and 30 minutes before a regular meal, preferably between 30 minutes and 60 minutes before a regular meal, more preferably between 60 minutes and 90 minutes, preferably between 90 minutes and 120 minutes, preferably between 120 minutes and 150 minutes, preferably between 150 minutes and 180 minutes before a regular meal.

[0477] Preferred numbered embodiment 22. An AWA or a dosage form containing an effective amount of an AWA for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or dosage form is administered as a pre-meal meal, such as between 1 minute and 180 minutes before a regular meal.

[0478] Preferred numbered embodiment 23. AWA or a dosage form containing an effective amount of AWA for treatment and / or prevention according to any of the aforementioned preferred numbered embodiments, wherein the AWA or the dosage form containing an effective amount of AWA is administered in an amount sufficient to provide an AWA dose of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose.

[0479] Preferred numbered embodiment 24. AWA or a dosage form containing an effective amount of AWA for treatment and / or prevention according to any of the aforementioned preferred numbered embodiments, wherein the AWA or the dosage form containing an effective amount of AWA is administered in an amount sufficient to provide a dosage of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose, wherein the dose is administered between 1-5 times a day, such as 1, 2, 3, 4 or 5 times a day.

[0480] Preferred numbered embodiment 25. An AWA or dosage form for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in an amount sufficient to provide an AWA dosage of 0.05 g / kg-4 g / kg body weight per day, preferably 0.05 g / kg-2 g / kg body weight per day, preferably 0.1 g / kg-1 g / kg body weight per day, more preferably 0.2 g / kg-0.5 g / kg body weight per day.

[0481] Preferred numbered embodiment 26. A dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule and an edible strip.

[0482] Preferred numbered embodiment 27. A dosage form for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form is a liquid, wherein the liquid is a "ready-to-drink beverage".

[0483] Preferred numbered embodiment 28. A dosage form for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0484] Preferred numbered embodiment 29. The dosage form for treatment and / or prevention according to any of the aforementioned preferred numbered embodiments, comprising AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

[0485] Preferred numbered embodiment 30. A dosage form for treatment and / or prevention according to any of the aforementioned preferred numbered embodiments, comprising AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

[0486] Preferred numbered embodiment 31. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising AWA in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0487] Preferred numbered embodiment 32. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising AWA in an amount of 1 g / 100 mL-20 g / 100 mL, more preferably 2 g / 100 mL-18 g / 100 mL, even more preferably at least 3 g / 100 mL-14 g / 100 mL and most preferably 5 g / 100 mL-10 g / 100 mL.

[0488] Preferred numbered embodiment 33. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0489] Preferred numbered embodiment 34. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL and most preferably 4 g / 100 mL-14 g / 100 mL.

[0490] Preferred numbered embodiment 35. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g and most preferably at least 50 g / 100 g.

[0491] Preferred numbered embodiment 36. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising AWA in an amount of at least 50 g / 100 g, more preferably at least 60 g / 100 g, more preferably at least 70 g / 100 g, even more preferably at least 80 g / 100 g and most preferably at least 90 g / 100 g.

[0492] Preferred numbered embodiment 37. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, comprising AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g and most preferably 30g / 100g-75g / 100g.

[0493] Preferred numbered embodiment 38. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, having a pH in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5.

[0494] Preferred numbered embodiment 39. The dosage form for treatment and / or prevention according to any of the preceding preferred numbered embodiments, having an acidic pH, preferably having a pH in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0495] Preferred numbered embodiments 40. A dosage form for use in treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form additionally comprises at least one non-dairy component.

[0496] Preferred numbered embodiments 41. A nutritional powder comprising

[0497] - AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein, and

[0498] - At least one non-dairy ingredient.

[0499] Preferred numbered embodiment 42. AWA for treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, or a nutritional powder comprising AWA according to any of the preceding preferred numbered embodiments, wherein the protein of the AWA comprises beta-lactoglobulin (BLG) in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and more preferably at least 90% w / w.

[0500] Preferred numbered embodiments 43. AWA for use in treatment and / or prevention or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, or a nutritional powder comprising AWA according to any of the preceding preferred numbered embodiments, wherein the AWA is obtainable by, and for example by:

[0501] a. Providing a whey protein solution, the whey protein solution:

[0502] -Having a pH in the range of 6-9

[0503] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0504] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0505] - Contains at least 30% BLG relative to total protein

[0506] b. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0507] c. subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, preferably freeze drying, followed by particle size reduction, thereby obtaining agglomerates of soluble whey protein aggregates.

[0508] Preferred numbered embodiment 44. A method of producing a composition comprising AWA, comprising the steps of

[0509] a. Providing a whey protein solution, the whey protein solution:

[0510] -Having a pH in the range of 6-9

[0511] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0512] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0513] - Contains at least 30% BLG relative to total protein

[0514] b. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0515] c. subjecting the suspension or a protein concentrate thereof to a process resulting in agglomeration of the sWPA, preferably freeze drying, and preferably subsequently reducing the particle size of the freeze dried product, thereby obtaining a composition comprising agglomerates of soluble whey protein aggregates.

[0516] Preferred numbered embodiments 45. A method for treating and / or preventing one or more conditions associated with GLP-1 blood levels, wherein the method comprises administering AWA or a dosage form containing an effective amount of AWA to a human subject.

[0517] Preferred numbered embodiment 46. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the condition is selected from one or more of the following:

[0518] a. Type II diabetes

[0519] b. Prediabetes

[0520] c. Hyperglycemia

[0521] d. Non-alcoholic fatty liver disease (NAFLD)

[0522] e. Nonalcoholic steatohepatitis (NASH)

[0523] f. Overweight

[0524] g. Obesity

[0525] h. Obesity-related diseases

[0526] i. Muscle atrophy

[0527] j. Cardiovascular disease

[0528] k. Arteriosclerosis

[0529] 1. Hypertension

[0530] m.Dyslipidemia.

[0531] Preferred numbered embodiments 47. A method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA is expressed in a human subject.

[0532] a. Increase satiety

[0533] b. Reduce hunger

[0534] c. Lose weight

[0535] d. Improve blood sugar control

[0536] e. Reduce glucagon secretion

[0537] f. Enhance insulin secretion

[0538] g. Reduce body and / or liver fat

[0539] h. Improve muscle atrophy, and / or

[0540] i. Reduce the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

[0541] Preferred numbered embodiment 48. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the human subject is an adult, and more preferably an elderly person.

[0542] Preferred numbered embodiment 49. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the human subject is a young adult.

[0543] Preferred numbered embodiment 50. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the human subject is overweight and more preferably obese.

[0544] Preferred numbered embodiments 51. The method for treatment and / or prevention according to any one of the preceding preferred numbered embodiments, wherein said AWA or said dosage form is administered by oral administration.

[0545] Preferred numbered embodiment 52. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in combination with a meal, preferably as a pre-meal meal, between 5 minutes and 30 minutes before a regular meal, preferably between 30 minutes and 60 minutes before a regular meal, more preferably between 60 minutes and 90 minutes, preferably between 90 minutes and 120 minutes, preferably between 120 minutes and 150 minutes, preferably between 150 minutes and 180 minutes before a regular meal.

[0546] Preferred numbered embodiment 53. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or dosage form is administered as a pre-meal meal, such as between 1 minute and 180 minutes before a regular meal.

[0547] Preferred numbered embodiment 54. A method for treatment and / or prevention according to any of the aforementioned preferred numbered embodiments, wherein the AWA or a dosage form containing an effective amount of the AWA is administered in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose.

[0548] Preferred numbered embodiment 55. A method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or a dosage form containing an effective amount of the AWA is administered in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose, wherein the dose is administered between 1-5 times per day, such as 1, 2, 3, 4 or 5 times per day.

[0549] Preferred numbered embodiment 56. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-4 g / kg body weight per day, preferably 0.05 g / kg-2 g / kg body weight per day, preferably 0.1 g / kg-1 g / kg body weight per day, more preferably 0.2 g / kg-0.5 g / kg body weight per day.

[0550] Preferred numbered embodiment 57. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule, and an edible bar.

[0551] Preferred numbered embodiment 58. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form is a liquid, wherein the liquid is a "ready-to-drink beverage".

[0552] Preferred numbered embodiment 59. A method for treatment and / or prevention according to any one of the preceding preferred numbered embodiments, wherein the dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0553] Preferred numbered embodiment 60. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

[0554] Preferred numbered embodiment 61. A method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

[0555] Preferred numbered embodiment 62. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0556] Preferred numbered embodiment 63. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 1 g / 100 mL - 20 g / 100 mL, more preferably 2 g / 100 mL - 18 g / 100 mL, even more preferably at least 3 g / 100 mL - 14 g / 100 mL and most preferably 5 g / 100 mL - 10 g / 100 mL.

[0557] Preferred numbered embodiment 64. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0558] Preferred numbered embodiment 65. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL and most preferably 4 g / 100 mL-14 g / 100 mL.

[0559] Preferred numbered embodiment 66. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g and most preferably at least 50 g / 100 g.

[0560] Preferred numbered embodiment 67. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 50 g / 100 g, more preferably at least 60 g / 100 g, more preferably at least 70 g / 100 g, even more preferably at least 80 g / 100 g and most preferably at least 90 g / 100 g.

[0561] Preferred numbered embodiment 68. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g and most preferably 30g / 100g-75g / 100g.

[0562] Preferred numbered embodiment 69. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5.

[0563] Preferred numbered embodiment 70. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is acidic, preferably in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0564] Preferred numbered embodiments 71. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the dosage form additionally comprises at least one non-dairy component.

[0565] Preferred numbered embodiment 72. The method for treatment and / or prevention according to any of the preceding preferred numbered embodiments, wherein the protein of AWA comprises beta-lactoglobulin (BLG) in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and more preferably at least 90% w / w.

[0566] Preferred numbered embodiment 73. A method for treatment and / or prevention according to any one of the preceding preferred numbered embodiments, wherein AWA is obtainable by, and for example by:

[0567] a. Providing a whey protein solution, the whey protein solution:

[0568] -Having a pH in the range of 6-9

[0569] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0570] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0571] - Contains at least 30% BLG relative to total protein

[0572] b. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0573] c. subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, preferably freeze drying, followed by particle size reduction, thereby obtaining agglomerates of soluble whey protein aggregates.

[0574] Preferred Numbered Embodiments 74. A non-therapeutic method for treating a human subject with

[0575] a. Increase and / or prolong the secretion of glucagon-like peptide 1 (GLP-1)

[0576] b. Provide a biphasic secretion profile of GLP-1

[0577] c. Providing a delayed secretion profile of GLP-1

[0578] d. Increase satiety

[0579] e. Reduce hunger

[0580] f. Lose weight

[0581] g. Improve blood sugar control

[0582] h. Reduce glucagon secretion

[0583] i. Enhance insulin secretion

[0584] j. Reduce body and / or liver fat

[0585] k. Improve muscle atrophy, and / or

[0586] l. Reduce the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events,

[0587] The method comprises administering to a human subject an effective amount of AWA or a dosage form containing an effective amount of AWA.

[0588] Preferred numbered embodiment 75. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the human subject is an adult, and more preferably an elderly.

[0589] Preferred numbered embodiment 76. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the human subject is a young adult.

[0590] Preferred numbered embodiment 77. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the human subject is overweight and more preferably obese.

[0591] Preferred numbered embodiment 78. The non-therapeutic method according to any one of the preceding preferred numbered embodiments, wherein said AWA or said dosage form is administered by oral administration.

[0592] Preferred numbered embodiment 79. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in combination with a meal, preferably as a pre-meal meal, between 5 minutes and 30 minutes before a regular meal, preferably between 30 minutes and 60 minutes before a regular meal, more preferably between 60 minutes and 90 minutes, preferably between 90 minutes and 120 minutes, preferably between 120 minutes and 150 minutes, preferably between 150 minutes and 180 minutes before a regular meal.

[0593] Preferred numbered embodiment 80. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the AWA or dosage form is administered as a pre-meal meal, such as between 1 minute and 180 minutes before a regular meal.

[0594] Preferred numbered embodiment 81. A non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the AWA or a dosage form containing an effective amount of the AWA is administered in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose.

[0595] Preferred numbered embodiment 82. A non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the AWA or a dosage form containing an effective amount of the AWA is administered in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose, wherein the dose is administered between 1-5 times per day, such as 1, 2, 3, 4 or 5 times per day.

[0596] Preferred numbered embodiment 83. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-4 g / kg body weight per day, preferably 0.05 g / kg-2 g / kg body weight per day, preferably 0.1 g / kg-1 g / kg body weight per day, more preferably 0.2 g / kg-0.5 g / kg body weight per day.

[0597] Preferred numbered embodiments 84. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule, and an edible bar.

[0598] Preferred numbered embodiments 85. The non-therapeutic method according to any one of the preceding preferred numbered embodiments, wherein the dosage form is a liquid, wherein the liquid is a "ready-to-drink beverage".

[0599] Preferred numbered embodiment 86. The non-therapeutic method according to any one of the preceding preferred numbered embodiments, wherein the dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0600] Preferred numbered embodiment 87. A non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

[0601] Preferred numbered embodiment 88. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

[0602] Preferred numbered embodiment 89. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0603] Preferred numbered embodiment 90. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 1 g / 100 mL - 20 g / 100 mL, more preferably 2 g / 100 mL - 18 g / 100 mL, even more preferably at least 3 g / 100 mL - 14 g / 100 mL and most preferably 5 g / 100 mL - 10 g / 100 mL.

[0604] Preferred numbered embodiments 91. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0605] Preferred numbered embodiment 92. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL and most preferably 4 g / 100 mL-14 g / 100 mL.

[0606] Preferred numbered embodiment 93. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g and most preferably at least 50 g / 100 g.

[0607] Preferred numbered embodiment 94. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 50 g / 100 g, more preferably at least 60 g / 100 g, more preferably at least 70 g / 100 g, even more preferably at least 80 g / 100 g and most preferably at least 90 g / 100 g.

[0608] Preferred numbered embodiment 95. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g and most preferably 30g / 100g-75g / 100g.

[0609] Preferred numbered embodiment 96. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5.

[0610] Preferred numbered embodiments 97. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is acidic, preferably in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0611] Preferred numbered embodiments 98. The non-therapeutic method according to any one of the preceding preferred numbered embodiments, wherein the dosage form additionally comprises at least one non-dairy component.

[0612] Preferred numbered embodiment 99. The non-therapeutic method according to any of the preceding preferred numbered embodiments, wherein the protein of AWA comprises beta-lactoglobulin (BLG) in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and more preferably at least 90% w / w.

[0613] Preferred numbered embodiment 100. A non-therapeutic method according to any one of the preceding preferred numbered embodiments, wherein AWA is obtainable by, and for example by:

[0614] d. providing a whey protein solution, wherein the whey protein solution:

[0615] -Having a pH in the range of 6-9

[0616] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0617] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0618] - Contains at least 30% BLG relative to total protein

[0619] e. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C to 180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0620] f. subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, preferably freeze drying, followed by particle size reduction, thereby obtaining agglomerates of soluble whey protein aggregates.

[0621] Preferred numbered embodiments 101. An effective amount of AWA or a dosage form containing an effective amount of AWA for use in a human subject:

[0622] a. Increase and / or prolong the secretion of glucagon-like peptide 1 (GLP-1)

[0623] b. Provide a biphasic secretion profile of GLP-1

[0624] c. Providing a delayed secretion profile of GLP-1

[0625] d. Increase satiety

[0626] e. Reduce hunger

[0627] f. Lose weight

[0628] g. Improve blood sugar control

[0629] h. Reduce glucagon secretion

[0630] i. Enhance insulin secretion

[0631] j. Reduce body and / or liver fat

[0632] k. Improve muscle atrophy, and / or

[0633] l. Reduce the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

[0634] Preferred numbered embodiment 102. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the human subject is an adult, and more preferably an elderly person.

[0635] Preferred numbered embodiment 103. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the human subject is a young adult.

[0636] Preferred numbered embodiment 104. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the human subject is overweight and more preferably obese.

[0637] Preferred numbered embodiment 105. Use of an effective amount of an AWA or a dosage form containing an effective amount of an AWA according to any of the preceding preferred numbered embodiments, wherein said AWA or said dosage form is administered to a human subject by oral administration.

[0638] Preferred numbered embodiment 106. Use of an effective amount of an AWA or a dosage form containing an effective amount of an AWA according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered to a human subject in combination with a meal, preferably as a pre-meal meal, between 5 minutes and 30 minutes before a regular meal, preferably between 30 minutes and 60 minutes before a regular meal, more preferably between 60 minutes and 90 minutes, preferably between 90 minutes and 120 minutes, preferably between 120 minutes and 150 minutes, preferably between 150 minutes and 180 minutes before a regular meal.

[0639] Preferred numbered embodiment 107. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA or dosage form is administered to a human subject as a pre-meal meal, such as between 1 minute and 180 minutes before a regular meal.

[0640] Preferred numbered embodiment 108. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form containing an effective amount of AWA is administered to a human subject in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose.

[0641] Preferred numbered embodiment 109. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein AWA or a dosage form containing an effective amount of AWA is administered to a human subject in an amount sufficient to provide a dose of AWA of at least 0.05 g / kg lean body weight per dose, preferably at least 0.1 g / kg lean body weight per dose, more preferably at least 0.3 g / kg lean body weight per dose, even more preferably at least 0.5 g / kg lean body weight per dose, wherein the dose is administered between 1-5 times per day, such as 1, 2, 3, 4 or 5 times per day.

[0642] Preferred numbered embodiment 110. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA or the dosage form is administered to a human subject in an amount sufficient to provide a dosage of AWA of 0.05 g / kg-4 g / kg body weight per day, preferably 0.05 g / kg-2 g / kg body weight per day, preferably 0.1 g / kg-1 g / kg body weight per day, more preferably 0.2 g / kg-0.5 g / kg body weight per day.

[0643] Preferred numbered embodiments 111. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule, and an edible bar.

[0644] Preferred numbered embodiments 112. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form is a liquid, wherein the liquid is a "ready-to-drink beverage".

[0645] Preferred numbered embodiments 113. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0646] Preferred numbered embodiment 114. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

[0647] Preferred numbered embodiment 115. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

[0648] Preferred numbered embodiment 116. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0649] Preferred numbered embodiment 117. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 1 g / 100 mL-20 g / 100 mL, more preferably 2 g / 100 mL-18 g / 100 mL, even more preferably at least 3 g / 100 mL-14 g / 100 mL and most preferably 5 g / 100 mL-10 g / 100 mL.

[0650] Preferred numbered embodiment 118. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0651] Preferred numbered embodiment 119. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL and most preferably 4 g / 100 mL-14 g / 100 mL.

[0652] Preferred numbered embodiment 120. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g and most preferably at least 50 g / 100 g.

[0653] Preferred numbered embodiment 121. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 50g / 100g, more preferably at least 60g / 100g, more preferably at least 70g / 100g, even more preferably at least 80g / 100g and most preferably at least 90g / 100g.

[0654] Preferred numbered embodiment 122. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g and most preferably 30g / 100g-75g / 100g.

[0655] Preferred numbered embodiment 123. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5.

[0656] Preferred numbered embodiment 124. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is acidic, preferably in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0657] Preferred numbered embodiment 125. Use of a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the dosage form additionally comprises at least one non-dairy component.

[0658] Preferred numbered embodiment 126. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the protein of AWA comprises beta-lactoglobulin (BLG) in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and more preferably at least 90% w / w.

[0659] Preferred numbered embodiment 127. Use of an effective amount of AWA or a dosage form containing an effective amount of AWA according to any of the preceding preferred numbered embodiments, wherein the AWA is obtainable by, and for example by:

[0660] a. Providing a whey protein solution, the whey protein solution:

[0661] -Having a pH in the range of 6-9

[0662] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0663] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0664] - Contains at least 30% BLG relative to total protein

[0665] b. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0666] c. subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, preferably freeze drying, followed by particle size reduction, thereby obtaining agglomerates of soluble whey protein aggregates.

[0667] Preferred numbered embodiments 128. Use of AWA or a dosage form containing an effective amount of AWA in the preparation of a medicament for the treatment and / or prevention of one or more of the following

[0668] a. Type II diabetes

[0669] b. Prediabetes

[0670] c. Hyperglycemia

[0671] d. Non-alcoholic fatty liver disease (NAFLD)

[0672] e. Nonalcoholic steatohepatitis (NASH)

[0673] f. Overweight

[0674] g. Obesity

[0675] h. Obesity-related diseases

[0676] i. Muscle atrophy

[0677] j. Cardiovascular disease

[0678] k. Arteriosclerosis

[0679] 1. Hypertension

[0680] m.Dyslipidemia.

[0681] Preferred numbered embodiment 129. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule, and an edible bar.

[0682] Preferred numbered embodiment 130. Use of a dosage form comprising an effective amount of AWA for the preparation of a medicament according to any one of the preceding preferred numbered embodiments, wherein the dosage form is a liquid, wherein the liquid is a "ready-to-drink beverage".

[0683] Preferred numbered embodiments 131. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any one of the preceding preferred numbered embodiments, wherein the dosage form is a powder intended to be dissolved in a liquid prior to ingestion.

[0684] Preferred numbered embodiment 132. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

[0685] Preferred numbered embodiment 133. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

[0686] Preferred numbered embodiment 134. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0687] Preferred numbered embodiment 135. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 1 g / 100 mL-20 g / 100 mL, more preferably 2 g / 100 mL-18 g / 100 mL, even more preferably at least 3 g / 100 mL-14 g / 100 mL and most preferably 5 g / 100 mL-10 g / 100 mL.

[0688] Preferred numbered embodiment 136. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of at least 1 g / 100 mL, preferably at least 2 g / 100 mL, preferably at least 4 g / 100 mL, preferably at least 6 g / 100 mL, preferably at least 8 g / 100 mL, preferably at least 10 g / 100 mL, preferably at least 12 g / 100 mL, preferably at least 14 g / 100 mL, preferably at least 16 g / 100 mL, preferably at least 18 g / 100 mL, preferably at least 20 g / 100 mL, preferably at least 22 g / 100 mL, preferably at least 24 g / 100 mL, preferably at least 26 g / 100 mL, preferably at least 28 g / 100 mL, preferably at least 30 g / 100 mL.

[0689] Preferred numbered embodiment 137. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL and most preferably 4 g / 100 mL-14 g / 100 mL.

[0690] Preferred numbered embodiment 138. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g and most preferably at least 50 g / 100 g.

[0691] Preferred numbered embodiment 139. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of at least 50 g / 100 g, more preferably at least 60 g / 100 g, more preferably at least 70 g / 100 g, even more preferably at least 80 g / 100 g and most preferably at least 90 g / 100 g.

[0692] Preferred numbered embodiment 140. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the dosage form comprises AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g and most preferably 30g / 100g-75g / 100g.

[0693] Preferred numbered embodiment 141. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.5.

[0694] Preferred numbered embodiment 142. Use of a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the pH of the dosage form is acidic, preferably in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.7.

[0695] Preferred numbered embodiment 143. Use of a dosage form containing an effective amount of AWA for the preparation of a medicament according to any one of the preceding preferred numbered embodiments, wherein the dosage form additionally comprises at least one non-dairy component.

[0696] Preferred numbered embodiment 144. Use of AWA or a dosage form containing an effective amount of AWA in the preparation of a medicament according to any of the preceding preferred numbered embodiments, wherein the protein of AWA comprises beta-lactoglobulin (BLG) in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and more preferably at least 90% w / w.

[0697] Preferred numbered embodiment 145. Use of AWA or a dosage form containing an effective amount of AWA in the preparation of a medicament according to any one of the preceding preferred numbered embodiments, wherein AWA is obtainable by, and for example, by:

[0698] a. Providing a whey protein solution, the whey protein solution:

[0699] -Having a pH in the range of 6-9

[0700] - has a weight ratio between total protein and total calcium and magnesium content of at least 100

[0701] - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution

[0702] - Contains at least 30% BLG relative to total protein

[0703] b. subjecting the whey protein solution to a heat treatment involving heating it to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG to obtain a suspension containing sWPA, and

[0704] c. subjecting the suspension or its protein concentrate to a process leading to agglomeration of the sWPA, preferably freeze drying, followed by particle size reduction, thereby obtaining agglomerates of soluble whey protein aggregates. Example

[0705] Example 1: Agglomerates of soluble whey protein aggregates

[0706] 1.1 Preparation of AWA by freeze drying

[0707] Agglomerates of soluble whey protein aggregates (AWA) were prepared by freeze drying a suspension of soluble whey protein aggregates prepared from whey protein isolate powder (SPI) obtained from natural milk whey proteins. The chemical composition of SPI is summarized in Table 1: Composition of whey protein isolate (SPI) obtained from natural milk whey proteins used to prepare agglomerates of soluble whey protein aggregates.

[0708]

[0709]

[0710] Prepare a suspension of soluble whey protein aggregates by the following procedure:

[0711] - 37.8 kg of demineralized water were transferred to the Scanima mixer,

[0712] - Heat the softened water to 45℃-50℃,

[0713] - 2.2 kg of SPI powder prepared from milk whey was added to demineralized water and dissolved by slow stirring for 1 hour,

[0714] - Adjust the pH of the solution to pH 7.4 by slowly adding 6% NaOH in small doses,

[0715] - the pH adjusted solution was then heated to 80°C and maintained at this temperature for 20 minutes to produce soluble whey protein aggregates,

[0716] - After a holding time of 20 minutes, the temperature is lowered to about 6 degrees by indirect cooling,

[0717] - Place the cooled suspension in a plastic tray stored at -20°C to -25°C,

[0718] - The frozen suspension is then freeze-dried under vacuum for 24 hours (about 1 mBar, starting at -40°C and gradually adding heating), which causes the frozen water to sublime into steam and produces a dry product,

[0719] - The dried product is finally milled to form a powder containing agglomerates of soluble whey protein aggregates.

[0720] The content of agglomerated soluble whey protein aggregates present after the freeze-drying process was estimated to be 80% w / w relative to the total protein.

[0721] Table 1: Composition of whey protein isolate (SPI) obtained from native milk whey protein for the preparation of agglomerates of soluble whey protein aggregates

[0722]

[0723]

[0724] *) Below detection limit

[0725] The gelling properties of the agglomerates (AWA-f) prepared in Example 1.1 were compared with soluble whey protein aggregates (sWPA) by acidification with glucono-delta-lactone (GDL).

[0726] The samples for acidification were prepared by the following procedure: 20 g of AWA-f powder was mixed with demineralized water to a total of 500 g to obtain 4% protein. The solution was placed in a refrigerator to hydrate overnight.

[0727] A 4% sWPA solution was prepared by diluting 5% sWPA in demineralized water (prepared on the same day).

[0728] Acidification and simultaneous gel strength measurements were performed as follows: 200 g of sample was placed in a water bath equilibrated at 42°C. 5 g of GDL was added to give 2.5% GDL while stirring and pH was recorded: The solution was first stirred for 1 minute to ensure mixing into the sample. 19.6 mL was then transferred to the rheometer (Anton Paar with CC27 setup) and the measurement started. Rheometer settings: 1 Hz, 0.05%, 1 minute per point, 1 hour at 42°C (first two points not measured)

[0729] The rest of the sample was incubated in a water bath (without stirring) and the pH was measured every 1 minute using a pH recorder. After the measurement, the pH and gel strength (G', storage modulus) were correlated.

[0730] The sWPA samples showed a rapid increase in gel strength at the beginning of acidification, but then the gel dissolved at lower pH as shown by a decrease in gel strength.

[0731] The AWA-f sample showed a delayed increase in gel strength, ending with high gel strength at lower pH values, eventually exceeding the gel strength of sWPA at a pH less than about 4.

[0732] 1.2 Calcium-induced AWA formation

[0733] Six samples of calcium-induced AWA (AWA-C) were prepared from a suspension of soluble whey protein aggregates (sWPA) prepared from whey protein isolate powder (SPI) obtained from native milk whey proteins as disclosed in Example 1.1.

[0734] Six samples were prepared to have calcium:protein molar ratios of 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, and 5:1, and the samples were named AWA-C2.5, AWA-C3, AWA-C3.5, AWA-C4, AWA-C4.5, and AWA-C5, respectively.

[0735] More specifically, AWA-C samples were prepared by taking samples of 40 mL of sWPA suspension and equilibrating them at 75° C. in a water bath. 10 mL of CaCl2 solution (each sWPA sample was mixed with calcium solution to give the final mixture the desired molar ratio between calcium and protein) was then added, while stirring, ending with a final protein concentration of 4% w / w. The mixture was incubated at 75° C. for 3 minutes with stirring. Subsequently, the mixture was cooled to 23° C. and allowed to incubate at this temperature for about 3 hours to provide samples AWA-C2.5 to AWA-C5, which were immediately analyzed.

[0736] Samples AWA-C2.5 to AWA-C5 were all liquid compositions with slight development of opacity, and AWA-C3.5 to AWA-C5 also contained visible agglomerates.

[0737] A larger sample of AWA-C5 (AWA-C5b) was further prepared in the same manner by equilibrating 160 mL of sWPA suspension at 75°C and adding 40 mL of CaCl2 solution under stirring to give a final molar stoichiometry of 5:1 calcium:protein.

[0738] The gelling properties and pH stability of agglomerates of AWA-C5b prepared by acidification with glucono-delta-lactone (GDL) were compared with sWPA suspension and with AWA-f suspension prepared from Example 1.1.

[0739] Samples were treated with two different GDL concentrations (1% w / w and 5% w / w) to enable analysis of samples at different final pH after incubation for 1 hour at 37°C.

[0740] Acidification was performed as follows: 3 40 g samples were taken from each of the sWPA suspension, AWA-f and AWA-C5b and equilibrated at 37°C (i.e., 9 samples in total). One of the three samples of each protein sample type was left untreated, and the other two samples were mixed with 0.4 g GDL (about 1% w / w) and 2 g GDL (about 5% w / w), respectively. The samples were stirred for 1 minute to evenly distribute the GDL. The samples were then incubated at 37°C for 1 hour without stirring. Subsequently, 20 g from each sample was centrifuged at 3000 g for 5 minutes at room temperature, after which the total protein content of the supernatant was determined according to analysis 7 of WO2021136785A1. In addition, the final pH was measured and found to be:

[0741] For samples to which GDL was not added, pH 7.1 ± 0.3

[0742] For samples to which 1% w / w GDL was added, pH 4.2 ± 0.1, and

[0743] For samples to which 5% w / w GDL was added, pH 2.9 ± 0.1.

[0744] The results of Example 1.2 have been plotted on Figure 2 As can be seen from the figure, the sWPA sample had all the protein in the supernatant after centrifugation at pH 7.1, but when it was acidified to pH 4.2, it had formed a gel and most of the protein was in the precipitate. However, when the pH reached 2.9, all the gel had dissolved and the protein was found in the supernatant again. This behavior is expected from the gel strength measurements described in Example 1.1.

[0745] Figure 2 It is also shown that for both AWA-f and AWA-C5b, at pH 7.1, the protein concentration in the supernatant is low because the AWA has precipitated during the centrifugation process. During the acidification process, only a small amount of protein is released to the supernatant. At pH 4.2, about 35% of the protein of AWA-f and AWA-C5b is in the supernatant, which means that the remaining 65% is in the centrifugal precipitate (and is therefore in the form of agglomerates). Even at pH 2.9, where all the protein of the sWPA sample is returned to the supernatant, the AWA-f and AWA-C5b samples still have about 40% in the form of agglomerates (and only about 60% in the supernatant). This shows that AWA prepared by both freeze drying (AWA-f) and by Ca-induced sWPA agglomeration (AWA-C5b) has better structural stability and is able to provide and maintain a higher gel strength than a comparable amount of sWPA under acidic conditions found in the human stomach.

[0746] High gel strength of gastric contents has previously been associated with increased satiety and fullness, and it is therefore apparent that effective amounts of AWA can be used to increase satiety, and thus can be used to treat and prevent, for example, obesity and obesity-related diseases and conditions. The GLP-1 boosting effect of AWA described in Example 2 further contributes to its medical usefulness.

[0747] Example 2: Physiological responses to AWA ingestion in human subjects

[0748] 2.1 Protein composition

[0749] The following protein ingredients (in powder form) were tested in vivo:

[0750] - The milled AWA-containing product obtained from Example 1.1 (at Figure 1A and Figure 1B AWA) is compared with

[0751] - The original SPI having the composition shown in Table 1 (in Figure 1A and Figure 1B SPI

[0752] - Protein hydrolysate of SPI prepared by enzymatic digestion to a degree of hydrolysis of about 23 (in Figure 1A and Figure 1B WPH

[0753] - Micellar casein isolate containing 86.5% total protein, 95% total casein of total protein, 5% whey protein of total protein, 1.5% fat, 0.4% lactose, 8.2% ash (in Figure 1A and Figure 1B MCI

[0754] 2.2 Subject characteristics

[0755] Ten healthy volunteers, five females and five males, were included in both the young group (young) and the old group (old). Exclusion criteria were chronic diseases, smoking, alcohol abuse, lactose intolerance, irregular menstrual cycle, and intake of drugs known to affect protein metabolism. Before the start of the clinical trial, the subjects underwent DEXA scanning to determine body composition. For the young and old groups, age, weight, height, BMI, and fat-free mass were 22.7±0.8 years and 76.3±1.5 years; 72±4 kg and 74±5 kg; 23.3±0.9 kg / m 2 and 25.8±0.9kg / m 2 ; 171±2cm and 169±4cm; 50±3kg and 47±3kg. Age range in the elderly group: 70.4-82.4 years old, young group: 18.2-25.0 years old.

[0756] Table 2: Subjects in the study (mean ± SEM)

[0757] female young male female elderly male Age (years) 23.6±0.6 22.7±0.8 21.8±1.4 77.2±2.0 76.3±1.5 74.2±2.1 Weight(kg) 65±4 72±4 79±4 64±3 74±5 88±5 BMI 22.2±1.5 23.3±0.9 24.5±0.9 24.0±0.8 25.8±0.9 28.1±1.0 height 171±2 175±2 180±3 163±2 169±4 177±5 FFM 42±1 50±3 57±4 40±2 47±3 58±1

[0758] 2.3 Experimental Day

[0759] Subjects were instructed to avoid vigorous physical activity two days before the experimental day. On the experimental day, subjects arrived on an empty stomach overnight, and were only allowed to drink water at will during the experimental day. When they arrived, they were placed on a hospital bed, and an intravenous catheter was placed in the antecubital vein; one in each arm was used for blood sampling and tracer infusion, respectively. Background blood samples were collected, and then the initiation of continuous infusion of different stable isotope labeled amino acid tracers purchased from Cambridge Isotope Laboratories (Andover, USA) was started. Blood samples were obtained 100 minutes and 115 minutes after the tracer infusion began. After obtaining 115 minutes of baseline samples, protein components (0.6g / kg fat-free mass) dissolved / dispersed in water were provided, and subjects were encouraged to take liquid solutions in about 5 minutes, and then the beaker glass was rinsed with 100ml water to remove the protein connected to the wall, and the subject was required to also take the 100ml water. Blood samples were collected at 15, 30, 45, 60, 90, 120, 180, 210, 240, 270, 300, and 360 minutes after the subjects began to consume the protein drink. Blood samples were collected into cooled EDTA tubes and centrifuged for 10 minutes. Plasma was transferred to Eppendorf tubes and stored at -80°C until analysis.

[0760] 2.4 Analysis

[0761] Plasma samples were analyzed at the Clinical Metabolomics Core Facility, (Rigshospitalet, Denmark) according to et al. (8) determined the concentration and enrichment of circulating amino acids. Hormones were determined by ELISA (Mercodia, Uppsala, Sweden) and total glucagon-like peptide 1 (GLP-1) was measured by luminescence.

[0762] Statistical analysis

[0763] Data are expressed as mean ± SEM. A two-way repeated measures ANOVA with time and protein as factors was used to compare the effects of milk protein composition dynamics over time. When significant interactions between factors were determined, Tukey's test was used to find time points where the effects of the components were significantly different. Statistical analysis was performed in OriginPro 2020. The statistical significance level was set at P < 0.05.

[0764] 2.6 Results

[0765] After ingestion of AWA, the GLP-1 plasma response was biphasic and exhibited a first rapid small peak at 30 minutes followed by a delayed / prolonged larger increase / peak. Notably, in young subjects, plasma GLP-1 was higher than other protein sources from 180 minutes to 360 minutes. In elderly subjects, this significantly higher GLP-1 after AWA was evident after 120 minutes.

[0766] For specific effects of AWA, the comparison with the native form (SPI) is most relevant.

[0767] In young subjects, the peak GLP-1 concentration (i.e., Cmax) after the AWA meal was about the same as after administration of WPH, but much higher than any peak for MCI and SPI. In elderly subjects, the peak GLP-1 concentration for AWA was about the same as any of WPH, MCI, or SPI. At the same time, the overall GLP-1 response (area under the curve) was increased and prolonged in both young and elderly subjects compared to any of WPH, MCI, and SPI.

[0768] As documented in Example 1.2, AWA prepared by, for example, Ca-induced agglomeration of sWPA has the same type of structural stability as AWA prepared by freeze drying (e.g., according to Example 1.1). Thus, it is apparent that the ability of AWA to increase and prolong the GLP-1 response in humans when orally administered is independent of how the AWA is produced.

[0769] References

[0770] Abdulla, H., Phillips, BE, Wilkinson, DJ, Limb, M., Jandova, T., Bass, JJ, Rankin, D., Cegielski, J., Sayda, M., Crossland, H., Williams, JP, Smith, K., Idris, I., & Atherton, PJ (2020). Glucagon-like peptide 1infusions overcome anabolicresistance to feeding in older human muscle.Aging Cell,19(9),1-12.doi.org / 10.1111 / acel.13202

[0771] A, van Hall G. Quantitative amino acid profiling and stable isotopically labeled amino acid tracer enrichment used for in vivo human systemic and tissue kinetics measurements. J Chromatogr B Analyt Technol Biomed Life Sci. 2014;951 - 952:69 - 77.

[0772] Cruz - Jentoft et al. 2019. Sarcopenia: revised European consensus on definition and diagnosis. Age and Ageing 2019;48:16 - 31. doi:10.1093 / ageing / afy169

[0773] Flint, A., Andersen, G., Hockings, P., Johansson, L., Morsing, A., Sundby Palle, M., Vogl, T., Loomba, R., & Plum - , L. (2021). Randomised clinical trial: semaglutide versus placebo reduced liver steatosis but not liver stiffness in subjects with non - alcoholic fatty liver disease assessed by magnetic resonance imaging. Alimentary Pharmacology and Therapeutics, 54(9), 1150 - 1161. doi.org / 10.1111 / apt.16608

[0774] Gurjar,A.A.,Kushwaha,S.,Chattopadhyay,S.,Das,N.,Pal,S.,China,S.P.,Kumar,H.,Trivedi,A.K.,Guha,R.,Chattopadhyay,N.,&Sanyal,S.(2020).Long actingGLP-1analog liraglutide ameliorates skeletal muscle atrophy inrodents.Metabolism:Clinical and Experimental,103,154044.doi.org / 10.1016 / j.metabol.2019.154044

[0775] Hong,Y.,Lee,J.H.,Jeong,K.W.,Choi,C.S.,&Jun,H.S.(2019).Amelioration ofmuscle wasting by glucagon-like peptide-1receptor agonist in muscleatrophy.Journal of Cachexia,Sarcopenia and Muscle,10(4),903-918.doi.org / 10.1002 / jcsm.12434

[0776] Kaneto,H.,Kimura,T.,Shimoda,M.,Obata,A.,Sanada,J.,Fushimi,Y.,Nakanishi,S.,Mune,T.,&Kaku,K.(2021).Favorable effects of GLP-1receptoragonist against pancreaticβ-cell glucose toxicity and the development ofarteriosclerosis:“the earlier,the better”in therapy with incretin-basedmedicine.International Journal of Molecular Sciences,22(15),1-11.doi.org / 10.3390 / ijms22157917

[0777] Krieger,J.P.(2020).Intestinal glucagon-like peptide-1effects on foodintake:Physiological relevance and emerging mechanisms.Peptides,131(June),170342.doi.org / 10.1016 / j.peptides.2020.170342

[0778] Love,K.M.,Liu,J.,Regensteiner,J.G.,Reusch,J.E.B.,&Liu,Z.(2020).GLP-1andinsulin regulation of skeletal and cardiac muscle microvascular perfusionin type 2 diabetes.Journal of Diabetes,12(7),488-498.doi.org / 10.1111 / 1753-0407.13045

[0779] Nguyen,T.T.N.,Choi,H.,&Jun,H.S.(2020).Preventive Effects ofDulaglutide on DisuseMuscle Atrophy Through Inhibition of Inflammation andApoptosis by Induction of Hsp72Expression.Frontiers in Pharmacology,11(February),1-11.doi.org / 10.3389 / fphar.2020.00090

[0780] Patel Chavez,C.,Cusi,K.,&Kadiyala,S.(2022).The Emerging Role ofGlucagon-likePeptide-1 Receptor Agonists for the Management of NAFLD.TheJournal of ClinicalEndocrinology and Metabolism,107(1),29-38.doi.org / 10.1210 / clinem / dgab578

[0781] Patel,V.J.,Joharapurkar,A.A.,Shah,G.B.&Jain,M.R.(2014)Effect of GLP-1basedtherapies on diabetic dyslipidemia.Curr Diabetes Rev 10(4):238-50.doi:10.2174 / 1573399810666140707092506.

[0782] Subaran,S.C.,Sauder,M.A.,Chai,W.,Jahn,L.A.,Fowler,D.E.,Aylor,K.W.,Basu,A.,&Liu,Z.(2014).GLP-1 at physiological concentrations recruits skeletaland cardiac musclemicrovasculature in healthy humans.Clinical Science,127(3),163-170.doi.org / 10.1042 / CS20130708

[0783] Zhao,Y.,Vanhoutte,P.M.,&Leung,S.W.S.(2015).Vascular nitric oxide:Beyond eNOS.Journal of Pharmacological Sciences,129(2),83-94.doi.org / 10.1016 / j.jphs.2015.09.002。

Claims

1. An agglomerate of soluble whey protein aggregates (AWA) or a dosage form containing an effective amount of AWA for use as a medicament.

2. AWA according to claim 1 or said dosage form containing an effective amount of AWA, for use in treating and / or preventing one or more conditions associated with GLP-1 blood levels in a human subject.

3. The AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the AWA promotes one or more of the following in a human subject: i. Increased satiety and / or reduced hunger ii. Weight loss iii. Improve blood sugar control iv. Decreased glucagon secretion v. Enhanced insulin secretion vi. Decreased body and / or liver fat vii. Improve muscle atrophy viii. Reduce the risk of arteriosclerosis, hypertension, dyslipidemia and / or cardiovascular events.

4. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2 or 3, wherein the disorder associated with the blood level of GLP-1 is selected from one or more of the following: a. Overweight b. Obesity c. Obesity-related diseases d. Type II diabetes e. Prediabetes f. Hyperglycemia g. Non-alcoholic fatty liver disease (NAFLD) h. Nonalcoholic steatohepatitis (NASH) i. Muscle atrophy j. Cardiovascular disease k. Arteriosclerosis 1. Hypertension m.Dyslipidemia.

5. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the one or more disorders are selected from one or more of overweight, obesity and obesity-related disorders.

6. The AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 5, wherein the AWA promotes increased satiety and / or decreased hunger in human subjects.

7. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the one or more conditions are selected from one or more of type II diabetes, prediabetes and hyperglycemia.

8. The AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 7, wherein the AWA promotes improved blood sugar control, reduced glucagon secretion and / or enhanced insulin secretion in a human subject.

9. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the one or more disorders are selected from one or more of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

10. The AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 9, wherein the AWA promotes the reduction of body and / or liver fat in human subjects.

11. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the one or more disorders is muscle atrophy.

12. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 11, wherein the AWA promotes increased muscle microvascular perfusion.

13. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 2, wherein the one or more conditions are selected from one or more of cardiovascular disease, arteriosclerosis, hypertension and dyslipidemia.

14. The AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to any one of claims 1 to 13, wherein the AWA or the dosage form containing an effective amount of AWA is for oral administration to the human subject.

15. The AWA or the dosage form containing an effective amount of AWA for use in treatment and / or prevention according to any one of claims 1 to 14, wherein the AWA or the dosage form containing an effective amount of AWA is administered as a pre-meal meal, such as between 1 minute and 180 minutes before a regular meal.

16. AWA for use in treatment and / or prevention or the dosage form containing an effective amount of AWA according to any one of claims 1 to 15, wherein the AWA or the dosage form containing an effective amount of AWA is administered in an amount sufficient to provide an AWA dose of at least 0.05 g / kg lean body weight per dose.

17. The AWA for treatment and / or prevention or the dosage form containing an effective amount of AWA according to claim 16, wherein the dose is administered between 1 and 5 times a day.

18. The dosage form for use containing an effective amount of AWA according to any one of claims 1-17, wherein the dosage form is selected from the group consisting of: a liquid, a powder, a gel, a capsule, and an edible bar.

19. The dosage form for use containing an effective amount of AWA according to any one of claims 1 to 18, comprising AWA in an amount of at least 30% w / w relative to total protein.

20. The dosage form containing an effective amount of AWA for use according to any one of claims 1 to 19, comprising AWA in an amount of at least 4 g per intended dose.

21. A dosage form comprising an effective amount of AWA, and preferably comprising AWA in an amount of at least 30% w / w relative to total protein.

22. The dosage form of claim 21, wherein the dosage form is selected from the group consisting of a liquid, a powder, a gel, a capsule, and an edible bar.

23. The dosage form according to claim 21 or 22, having a pH in the range of 5.5-9.0, more preferably 6.0-8.0, even more preferably 6.2-7.7, and most preferably 6.5-7.

5.

24. The dosage form according to claim 21 or 22, having a pH in the range of 2-5.4, more preferably 2.5-5.0, and most preferably 3.0-4.

7.

25. The dosage form according to any one of claims 21-24, comprising AWA in an amount of at least 50% w / w, even more preferably at least 60% w / w, and more preferably at least 70% w / w relative to total protein.

26. The dosage form according to any one of claims 21-25, comprising AWA in an amount of at least 4 g per intended dose, more preferably at least 6 g per intended dose, even more preferably at least 8 g per intended dose, and most preferably at least 10 g per intended dose.

27. The dosage form according to any one of claims 21 to 26, which is a liquid dosage form, preferably a ready-to-drink liquid.

28. The dosage form according to claim 27, which is a pasteurized or sterile liquid.

29. The dosage form according to claim 27 or 28, wherein the liquid dosage form comprises AWA in an amount of 1 g / 100 mL - 20 g / 100 mL, more preferably 2 g / 100 mL - 18 g / 100 mL, even more preferably at least 3 g / 100 mL - 14 g / 100 mL, and most preferably 5 g / 100 mL - 10 g / 100 mL.

30. The dosage form according to any one of claims 27-29, wherein the liquid dosage form comprises total protein in an amount of 1 g / 100 mL-25 g / 100 mL, more preferably 2 g / 100 mL-20 g / 100 mL, even more preferably at least 3 g / 100 mL-16 g / 100 mL, and most preferably 4 g / 100 mL-14 g / 100 mL.

31. The dosage form according to any one of claims 21 to 26, which is a powder, preferably a powder intended to be dissolved in a liquid prior to ingestion.

32. The dosage form according to claim 31, wherein the powder dosage form comprises AWA in an amount of at least 10 g / 100 g, more preferably at least 15 g / 100 g, more preferably at least 20 g / 100 g, even more preferably at least 30 g / 100 g, and most preferably at least 50 g / 100 g.

33. The dosage form according to claim 31 or 32, wherein the powder dosage form comprises AWA in an amount of at least 50g / 100g, more preferably at least 60g / 100g, more preferably at least 70g / 100g, even more preferably at least 80g / 100g, and most preferably at least 90g / 100g.

34. The dosage form according to any one of claims 31-33, wherein the powder dosage form comprises AWA in an amount of 10g / 100g-90g / 100g, more preferably 15g / 100g-85g / 100g, even more preferably at least 20g / 100g-80g / 100g, and most preferably 30g / 100g-75g / 100g.

35. The dosage form of any one of claims 31-34, wherein the powder dosage form is a nutritional powder comprising: - AWA in an amount of at least 30% w / w relative to total protein, more preferably at least 50% w / w relative to total protein, even more preferably at least 60% w / w relative to total protein, and more preferably at least 70% w / w relative to total protein, and - At least one non-dairy ingredient.

36. The dosage form according to any one of claims 31-35, wherein the powder dosage form is a nutritional powder comprising AWA and at least one non-dairy protein in an amount of at least 30% w / w relative to total protein.

37. The dosage form according to any one of claims 21-36, wherein the preparation of the dosage form involves mixing an AWA-containing composition obtainable by a process according to one or more of claims 41-61.

38. The dosage form according to any one of claims 21-37, wherein the AWA used for preparing the dosage form is provided by an AWA-containing composition obtainable by a process according to one or more of claims 41-61.

39. The AWA according to any one of claims 1-20 or the dosage form containing an effective amount of AWA, or the dosage form according to any one of claims 21-36, wherein the AWA is obtainable by, and for example by: a. Providing a whey protein solution, the protein solution: -Having a pH in the range of 6-9 - has a weight ratio between total protein and total calcium and magnesium content of at least 100 - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution - Contains at least 30% BLG relative to total protein b. subjecting the whey protein solution to a heat treatment to obtain a suspension containing sWPA, the heat treatment involving heating the whey protein solution to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG, and c. subjecting said suspension or a protein concentrate thereof to a process which causes agglomeration of said sWPA, thereby obtaining agglomerates of soluble whey protein aggregates.

40. The AWA for use or the dosage form containing an effective amount of AWA according to claim 39, wherein the processing in step c is freeze drying followed by particle size reduction of the freeze dried product to obtain agglomerates of soluble whey protein aggregates.

41. A method for producing a composition comprising AWA, comprising the steps of a. Providing a whey protein solution, the whey protein solution: -Having a pH in the range of 6-9 - has a weight ratio between total protein and total calcium and magnesium content of at least 100 - comprising at least 1%-10% w / w BLG relative to the weight of the whey protein solution - Contains at least 30% BLG relative to total protein b. subjecting the whey protein solution to a heat treatment to obtain a suspension containing sWPA, the heat treatment involving heating the whey protein solution to a temperature in the range of 68°C-180°C for a period of time sufficient to denature at least 25% w / w of the BLG, and c. subjecting said suspension or a protein concentrate thereof to a process resulting in agglomeration of said sWPA, thereby obtaining a composition comprising agglomerates of soluble whey protein aggregates.

42. The method of claim 41, wherein the processing of step c involves one or more of: - Freeze drying, - increasing the level of divalent metal cations and / or increasing the level of monovalent metal cations, - lowering the pH, - Concentrating, preferably by one or more of reverse osmosis, nanofiltration, ultrafiltration, microfiltration, evaporation and combinations thereof.

43. A method according to claim 41 or 42, wherein the processing of step c resulting in agglomeration of the sWPA involves freeze drying.

44. The method according to any one of claims 41 to 43, wherein the agglomeration step is followed by particle size reduction of the freeze-dried product.

45. A method according to any one of claims 41 to 44, wherein the processing of step c resulting in agglomeration of the sWPA involves the addition of divalent metal ions, preferably calcium and / or magnesium.

46. ​​The method according to any one of claims 41 to 45, wherein the added divalent metal ions are provided by water-soluble salts of calcium and / or magnesium, preferably CaCl2, MgCl2, CaSO4, MgSO4, CaCO3, MgCO3, calcium phosphate and / or magnesium phosphate.

47. The method of any one of claims 41-46, wherein the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of at least 1:1, more preferably at least 2:1, even more preferably at least 3:1, and most preferably at least 4:

1.

48. The method according to any one of claims 41-47, wherein the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of 1:1-100:1, more preferably 2:1-60:1, even more preferably 3:1-40:1, and more preferably 3:1-30:

1.

49. The method according to any one of claims 41-48, wherein the divalent metal ions are added in an amount sufficient to obtain a molar ratio between the total amount of divalent metal ions and the amount of total protein of 1:1-20:1, more preferably 2:1-16:1, even more preferably 3:1-12:1, and more preferably 3:1-10:

1.

50. The method according to any one of claims 41 to 49, wherein the divalent metal ion-rich liquid has a molar percentage of calcium ions relative to the total molar content of divalent metal ions of at least 50%, more preferably at least 70%, even more preferably at least 80%, and most preferably at least 90%.

51. The method according to any one of claims 41-50, wherein the liquid rich in divalent metal ions has a temperature during the incubation in the range of 10°C-100°C, more preferably 20°C-90°C, even more preferably 35°C-85°C, and most preferably 40°C-80°C.

52. The method according to any one of claims 41-50, wherein the liquid rich in divalent metal ions has a temperature in the range of 10°C-90°C, more preferably 15°C-85°C, even more preferably 20°C-85°C, and most preferably 20°C-80°C during the incubation.

53. The method according to any one of claims 41-52, wherein the liquid rich in divalent metal ions is incubated for at least 1 minute, more preferably at least 2 minutes, and most preferably at least 3 minutes.

54. The method according to any one of claims 41-53, wherein the liquid rich in divalent metal ions is incubated for at least 0.5 hours, more preferably at least 1 hour, and most preferably at least 2 hours.

55. The method according to any one of claims 41-54, wherein the liquid rich in divalent metal ions is incubated for 1 minute to 48 hours, more preferably 2 minutes to 36 hours, and most preferably 3 minutes to 24 hours.

56. The method according to any one of claims 41-55, wherein the liquid rich in divalent metal ions is incubated for 0.5 hours to 48 hours, more preferably 1 hour to 36 hours, and most preferably 2 hours to 24 hours.

57. The method according to any one of claims 41 to 56, wherein the liquid rich in divalent metal ions is agitated during the incubation, such as by stirring.

58. The method of any one of claims 41-57, wherein step c further involves subjecting the incubated divalent metal ion-rich liquid to one or more of the following: o a concentration step, preferably one or more of evaporation, reverse osmosis, nanofiltration, ultrafiltration and / or microfiltration, and o A drying step, preferably involving spray drying.

59. The method according to claim 58, wherein the drying step involves preheating the liquid to be dried to a temperature in the range of 50-80°C before the actual spraying which converts the liquid to be dried into droplets from which water evaporates.

60. The method according to claim 58 or 59, wherein the liquid to be dried has a weight percentage of AWA of 2%-10% w / w, more preferably 3%-9% w / w, and most preferably 4%-8% w / w relative to the weight of the liquid.

61. The method according to any one of claims 58 to 60, wherein the liquid to be dried has a weight percentage of AWA relative to total protein of at least 40% w / w, more preferably at least 50% w / w, even more preferably at least 60% w / w, and most preferably at least 70% w / w.

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