WO2021064107A1

WO2021064107A1 - Composition comprising soy and use thereof in the prevention and/or treatment of microvascular or lipoprotein related diseases, microcardiovascular diseases, atherosclerosis, hypertension, and in patients with such cardiovascular disorders suffering from alzheimer and dementia, overweight and obesity, type 2 diabetes, asthma and other disorders

Info

Publication number: WO2021064107A1
Application number: PCT/EP2020/077533
Authority: WO
Inventors: Lars HØIE
Original assignee: Norvia Nutrition Ltd.
Priority date: 2019-10-02
Filing date: 2020-10-01
Publication date: 2021-04-08

Abstract

The invention concerns soy protein, phytoestrogens, phospholipids, and dietary fibers and compositions thereof suitable for inter alia preventing, treating and/or alleviating microvascular or lipoprotein related diseases of the heart, vascular and central nervous system in patients with hypertension, obesity/overweight, type 2 diabetes, Alzheimer disease, Dementia and other disorders.

Description

COMPOSITION COMPRISING SOY AND USE THEREOF IN THE PREVENTION AND/OR TREATMENT OF MICROVASCULAR OR LIPOPROTEIN RELATED DISEASES, MICROCARDIOVASCULAR DISEASES, ATHEROSCLEROSIS, HYPERTENSION, AND IN PATIENTS WITH SUCH CARDIOVASCULAR DISORDERS SUFFERING FROM ALZHEIMER AND DEMENTIA, OVERWEIGHT AND OBESITY, TYPE 2 DIABETES, ASTHMA AND OTHER DISORDERS

FIELD OF THE INVENTION

The present invention relates to soy protein and phytoestrogens in the form of soy isoflavones and compositions thereof for use in preventing, treating and/or alleviating; microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system; menopausal symptoms or premenstrual syndrome; high blood pressure, elevated blood lipids and blood sugar levels; hormonal prostate, breast or colon cancers; inflammation of e.g. joints and tendons; pulmonary diseases such as airway inflammation, asthma, bronchitis and small airways diseases.

The present invention also pertains to the use of such compositions for improving the physical performance and/or athletic performance as measured in watts of a human or animal subject, or for improving vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles, of a human subject.

Use of a composition according to the present invention is particularly useful in preventing and/or reducing the influx of lipoproteins and/or cholesterol and/or triglycerides through the endothelium into the arterial wall and/or causing dilation of blood vessels and/or reducing restriction of blood vessels and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and inflammation in subjects at high risk of developing microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system or subjects already suffering from microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system such as subjects suffering from arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, hypertension, Alzheimer disease or Dementia, or diabetic or obese/overweight subjects. A composition according to the present invention is also useful in treating e.g. chronic sclerosis, hypertension, Alzheimer disease or Dementia, or diabetic or obese/overweight subjects. Furthermore, the present invention also relates to the use of these compositions as a medicament and/or in the manufacture of a medicament for treating a subject suffering from; menopausal symptoms or premenstrual syndrome; high blood pressure, elevated blood lipid and blood sugar levels; hormonal prostate, breast or colon cancers; inflammation of joints and tendons; pulmonary diseases, more particularly chronic obstructive pulmonary disease (COPD), inflammation of the airways, asthma, bronchoconstriction, bronchitis, and/or small airways disease.

The present invention also concerns use of a composition according to the present invention in the prevention and/or treatment of said diseases and disorders or in order to achieve such improvements. In addition, the present invention also provides methods for preventing, treating, prophylactically treating and/or alleviating by therapy said diseases and disorders or in order to achieve such improvements.

BACKGROUND OF THE INVENTION

Lipid metabolism involves biosynthesis and degradation of e.g. fatty acids, triglycerides, phospholipids, i.e. phosphoglycerides, and cholesterol. Ingested triglycerides are hydrolyzed in the small intestine and hydrolysis products are absorbed by the intestinal mucosa. Due to the relative insolubility of dietary lipids in water, lipid digestion and absorption is facilitated by the action of detergent substances such as bile acids secreted from the gallbladder. Bile acids are essential for lipid digestion and absorption through the intestinal mucosa.

Triglycerides and cholesterol synthesized in the liver are transported in the bloodstream to peripheral tissues by transport proteins called lipoproteins.

Lipoproteins are tiny vesicles coated by apoproteins, phospholipids and free cholesterol and with an interior consisting of the more hydrophobic lipids, cholesteryl esters and triglycerides. Apoproteins and lipoproteins are primarily synthesized in the liver. The lipoproteins are capable of performing an apoprotein mediated binding to a receptor on the surface of a cell into which the entire lipoprotein particle is taken up and further metabolized.

Several different families of lipoproteins have been characterized and are traditionally classified by their density as determined by centrifugation. A standard lipoprotein classification scheme may include in increasing order of density, very low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL).

VLDL contains approximately 60 to 65 percent triglycerides and 5 to 10 percent cholesterol, lecithin and protein. They are relatively large and function in the transport of triglycerides from the liver to tissue. LDL contains approximately 40 to 50 percent cholesterol and 10 to 15 percent triglycerides, lecithin and protein. They are somewhat smaller than VLDL and also function in the transport of cholesterol from the liver to tissue. HDL contains roughly 75 percent lecithin and protein, while the rest is made up of cholesterol and a small amount of triglycerides. They function in the transport of cholesterol from tissue to the liver and, as such, have the opposite function of LDL. Cholesterol esters cannot readily traverse cellular membranes and are taken up by cells in a receptor-mediated process. Once bound to the LDL receptor, the LDL- particle is internalized by means of endocytosis, and cholesterol and fatty acids are released and further metabolized.

Investigations of the LDL receptor mediated internalization of cholesterol have generated a better understanding of the relationship between dietary cholesterol, plasma cholesterol levels, and the condition of artherosclerosis. It is believed that the white blood cells that accumulate cholesterol at sites of arterial injury contain a receptor termed a scavenger receptor. Like the LDL receptor, this scavenger receptor acts by the mechanism of endocytosis and mediates internalization of various extracellular materials. However, the scavenger receptor is indiscriminatory and takes up many different types of extracellular materials including oxidized LDL particles containing cholesterol. In contrast to the LDL receptor, the scavenger receptor is not down-regulated by a high concentration of cholesterol in the cell.

In addition to the above-mentioned lipoproteins, the organism also contains a type of lipoproteins called chylomicrons. Chylomicrons contain 90 to 95 percent triglycerides and only a small amount of cholesterol, lecithin and protein, and they function in the transport of triglycerides from the small intestine to e.g. muscles, liver and heart.

Phosphoglycerides, i.e. phospholipids derived from glycerol, consists of a glycerol backbone, two fatty acid side chains, and a phosphorylated alcohol. The fatty acid chains in phosphoglycerides usually contain an even number of carbon atoms, typically between 14 and 24, with the 16- and 18-carbon fatty acids being the most common. In animals, including humans, the hydrocarbon chain in fatty acids is unbranched and may be saturated or unsaturated.

In the simplest phosphoglyceride, phosphatidate, the hydroxyl groups at C-1 and C-2 of glycerol are esterified to the carboxyl groups of two fatty acid chains, and the C-3 hydroxyl group of the glycerol backbone is esterified to phosphoric acid. Only small amounts of phosphatidate exist in the organism. However, it is a key intermediate in the biosynthesis of other phosphoglycerides and the major phosphoglycerides are derivatives of phosphatidate. In this respect the phosphate group of phosphatidate becomes esterified to the hydroxyl group of one of several alcohols. The common alcohol moieties of phosphoglycerides are serine, ethanolamine, choline, glycerol and inositol, and thus the principal phosphoglycerides include phosphatidyl serine, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl glycerol and phosphatidyl inositol.

The metabolism of cholesterol in the human organism is closely linked to the synthesis, transport and degradation of triglycerides. As well cholesterol as phosphoglycerides are essential lipid component in all mammalian cells. They are used to regulate the fluidity of cellular membranes and serve as precursors for certain hormones, signal molecules, vitamins and bile acids. Cholesterol is synthesized in the liver and is transported with the blood to peripheral tissues by lipoproteins. The liver has a dual function in the metabolism of cholesterol since it is capable of both synthesizing cholesterol and converting surplus cholesterol into bile acids. It is also capable of excreting cholesterol into the bile. Phospholipids have been speculated to be involved in the transport of triglycerides through the liver, especially during mobilization from adipose (fatty) tissue. Because of their high concentration in the cell membranes, it is most likely they are involved in the transport of hydrophobic constituents into and out of cells.

Bile acids have ampholytic characteristics and contain both hydrophobic and hydrophilic surfaces. This ampholytic character facilitates a bile acid mediated emulsification of lipids into micelles. The formation of micelles allows digestive attacks by water-soluble enzymes and facilitates lipid absorption through the mucosal cells of the intestine. Bile acids are secreted from the liver and stored in the gallbladder before being passed through the bile duct and into the intestine. Biosynthesis of bile acids represents a major metabolic fate of cholesterol and accounts for more than half of the approximately 800 g cholesterol that is normally metabolized per day in a normal adult. Even though bile acids in an amount of 400 mg are synthesized each day, significantly more than this amount is secreted into the intestine. Most of the bile acids that are secreted into the upper small intestine are absorbed in the lower small intestine and are recycled to the liver. The process of enterohepatic circulation may amount to as much as 20 to 30 g of bile acids per day. In contrast, daily elimination of bile acids in the feces amounts to just 0.5 g or less.

Cholesterol acts on three different levels of regulation of its own synthesis. Firstly, it suppresses endogenous cholesterol synthesis by inhibiting HMG-CoA reductase.

Secondly, it activates acyl-CoA:cholesterol acyltransferase (ACAT) which is involved in the synthesis of cholesterol esters from cholesterol and fatty acids bound to acyl-CoA. Thirdly, cholesterol regulates synthesis of the LDL receptor. Accordingly, a decreased synthesis of LDL receptors will ensure that a cell in which a sufficient amount of cholesterol is already present does not take up cholesterol. This may explain why excessive dietary cholesterol generates a rapid elevation of cholesterol levels in the blood.

The presence of increased amounts of cholesterol in the blood is known to be positively correlated to microvascular diseases of the heart, vascular and/or central nervous system, conditions commonly attributed to the deposition on the inner lining of an arterial wall of plaque in the form of cholesterol and fats. Arteriosclerosis is a common term for a group of conditions related to the arterial system and leading to an increased arterial wall thickness and a subsequent loss of elasticity. Three main groups of arteriosclerosis frequently referred to are atherosclerosis, Monckeberg’s mediasclerosis and arteriolosclerosis. Atherosclerosis is most frequently observed in the aorta and in the main arteries connected thereto, in the coronary arteries and in the arteries of the brain. Monckeberg’s mediasclerosis leads to a narrowing of the media of the arteries of the extremities, and arteriolosclerosis is related to a narrowing of the small arteries and arterioles causing hypertension. Other arteriosclerotic manifestations include hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperglycemia, hypertension, and hyperinsulinemia

One commonly occurring arterial condition is that of atherosclerotic cardiovascular disease. The condition may eventually progress through several stages. A normal structure of an artery is characterized by discrete focal numbers of adhering monocytes, some intimal foam cells, and some intimal smooth muscle cells, or intimal cell masses at bifurcations. A fatty streak may occur non-symptomatically and involve a layer of foam cells. As microvascular diseases of the heart, vascular and/or central nervous system progresses, the cells making up the inner wall of an artery will gradually start to harden due to the deposition of lipid and calcium and proliferation of smooth muscle cells, and the cells may eventually become degenerated. As the wall of an artery thickens, hardens, and lose its elasticity during arteriosclerosis, the blood vessels may develop twists and turns and become narrowed so that the heart must work harder to pump the usual amount of blood through the arteries. The condition may regress or it may evolve into a formation of e.g. fibrous plaque. Fibrous plaque is a slowly reversible condition that may develop further into a complicated lesion.

The cellular degeneration is likely to result in a fracture of the arterial wall which in turn leads to the formation of a deposit of calcium, platelet formation and a gradual development of scar tissue further contributing to both cellular degeneration and a substantially reduced elasticity of the arterial wall. Atherosclerosis characterized by a restricted flow of blood through a coronary artery may lead to the development of coronary heart disease. A complicated lesion of an artery is often symptomatic, hardly reversible and may, in severe cases such as thrombosis, be lethal. A decreased flow of blood through an artery may lead to the formation of blood clots and this may eventually lead to thrombosis. If a blood clot forms in a coronary artery, the interruption of the blood flow may result in the death of part of the heart muscle and cause the extremely painful chest pains associated with a heart attack.

Arteriosclerotic symptoms largely depend on the arteries and tissue affected. When arteriosclerosis occurs in the arteries leading to the brain, the decrease in blood flow and oxygen can cause mental confusion and personality changes. A stroke may occur, if an artery in the brain that has been weakened by a rupture or a blood clot prevents blood from flowing to the brain. This may possibly result in e.g. partial paralysis, loss of speech, and sometimes even death. A decrease in the flow of blood through the coronary arteries results in a shortage of oxygen to the heart muscle and causes chest pains and a painful condition called angina pectoris. Angina pectoris is usually caused by a narrowing or an obstruction of a coronary artery. An attack of angina pectoris may be caused by stress or result from physical activities that require an increased supply of blood to the heart. Although it is well established that cholesterol, lipids and lipoproteins all contribute to the progression of various arteriosclerotic conditions in diabetic and non-diabetic subjects alike, little is known about the causes of microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system. Hereditary conditions clearly play a role in some cases and several socio-economical and life style related factors such as smoking, hypertension, dietary habits and continual stress also contribute to the development of microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system.

There is at present no simple cure or medical treatment for microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system and doctors usually advise patients to follow a low fat diet, to stop smoking and to exercise regularly. Patients suffering from hypercholesterolemia may be classified into four risk groups: (i) manifest coronary artery disease, (ii) other forms of atherosclerotic vascular disease, (iii) other risk factors for coronary artery disease in the absence of established atherosclerotic cardiovascular disease, and (iv) isolated hypercholesterolemia in the absence of other risk factors. The recommended treatment regimen for risk group (iv) is to give general advice together with the laboratory results to those patients having a total cholesterol level of 5.0 - 6.4 mmol/l, without any further follow-up. To patients with cholesterol levels in the range of 6.5 - 7.9 mmol/l and LDL levels > 5.0 mmol/l or an LDL/HDL ratio > 5.0, only non- pharmacological treatment is offered.

Drug treatment of cardiovascular diseases may include the use of calcium channel blockers to expand the arteries so that blood can flow more freely, and anticoagulants to prevent blood clots from forming in diseased arteries. Some studies indicate that compounds such as acetylsalicylic acid and sulphinpyrazone, which may reduce and/or inhibit clotting by reducing platelet reactivity, may also prevent formation of a thrombus. In advanced cases, surgery to replace diseased blood vessels with grafts of healthy arteries may be necessary. Furthermore, an ether phospholipid, i.e. the 1- alkyl-2-acetyl analog of phosphatidyl choline, has been advocated to play an important role in various arteriosclerotic conditions in diabetic and non-diabetic subjects alike.

This compound, known as platelet-activating factor, apart from promoting the aggregation of platelets also causes the dilation of blood vessels even in very low concentration (0.1 nM). Also, various lipid-lowering drugs have been advocated, as some studies have shown or indicated that even for otherwise healthy patients suffering from mild or moderate hypercholesterolemia, coronary morbidity and mortality is reduced when they are treated with such lipid-lowering drugs. The most widely used lipid-lowering drugs in recent years have been statins, such as HMG-CoA-reductase-inhibitors, bile acid resins, fibrates, nicotinic acid derivatives and various fish oil concentrates with a high content of w-3-fatty acids.

Furthermore, also treatments using niacin alone or in combination with a statin has been advocated to have an effect on cardiovascular events, progression/regression of coronary lesions and hypercholesterolemia. However, the dosages currently applicable are well above the recommended daily intake values, i.e. 18-20 mg/day, and adverse side effects have been reported to arise, when using the amounts of niacin presently shown to have an effect, i.e. 500-2000 mg/day.

Various forms of cerebrovascular insufficiency such as reduced blood supply to the brain or disrupted microvascular integrity in cortical regions may occupy an initiating or intermediate position in the chain of events ending with a setback leading to neuronal damage and ultimately the development of incapacitating neurological diseases like dementia and Alzheimer’s disease. In addition, evidence is fast accumulating which indicates that Alzheimer's disease is indeed primarily a vascular disorder with neurodegenerative consequences rather than a neurodegenerative disorder with vascular consequences.

Thus, impaired functionality of the small vessels of the cerebrovascular system i.e. the small arteries, arterioles, venules, and capillaries of the brain, can lead to so-called small vessel disease (CSVD), the clinical manifestations of which include e.g. cognitive decline, dementia, Alzheimer’s disease or the like. In fact, CSVD is thought to be among the main causes of vascular cognitive impairment and is thought to account for about 45% of dementia cases.

Apart from an increase in severity of arteriolosclerosis with aging, risk factors for CSVD include hypertension, cigarette smoking, diabetes, atrial fibrillation, hyperhomocysteinemia, chronic kidney disease, high circulatory phosphate level and obesity. Thus, controlling or removing these known risk factors is important for the prevention of CSVD. Also, improving vasomotor reserve capacity and cerebral endothelial function by inhibiting cerebral vascular superoxide production, enhancing endothelial-derived Nitric Oxide bioavailability and decreasing oxidative stress, could protect against cerebral ischemia. Many cardiovascular risk factors are abnormally high in individuals suffering from type 2 diabetes and these individuals are consequently at an increased risk of developing various arteriosclerotic vascular diseases. At the time of diagnosis of type 2 diabetes, the existence of arteriosclerotic manifestations is already pronounced in many individuals and may include hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, hyperglycemia, hypertension, and hyperinsulinemia.

In type 2 diabetes, an impaired insulin secretion as well as decreased insulin sensitivity is present. As a result of this, glucose is present in excessive amounts in the bloodstream in association with either low, normal or even high insulin levels.

Serum levels of glucose vary quite significantly depending on the nutritional status of a subject. Following a dietary intake rich in glucose containing carbohydrates, several homeostatic mechanisms are capable of promoting glucose uptake into cells as well as facilitating the metabolism of glucose leading e.g. to the synthesis of glycogen in the liver and muscles. When glucose levels subsequently fall some time after a meal, other regulatory mechanisms initiate the release of glucose from glycogen and initiate gluconeogenesis in order to maintain the blood glucose levels within the required limits.

Some homeostatic mechanisms are dependent on the action of hormones, and the most important hormone promoting glucose uptake and metabolism is insulin. In contrast, other hormones such as e.g. glucagon and epinephrine act antagonistically and facilitate increased blood glucose levels. Insulin is synthesized in pancreatic b cells and secreted in response to e.g. increased levels of blood glucose.

Once secreted into the bloodstream, insulin acts in several processes to promote i) uptake of metabolizable substrates into certain cells, ii) storage of lipids and glycogen, and iii) biosynthesis of macromolecules such as nucleic acids and proteins. More specifically, the action of insulin results in i) an increased uptake of glucose in muscles and adipose tissues, ii) activation of the glycolytic pathway in the liver, iii) an increased synthesis of fatty acids and triglycerides in the liver and adipose tissues, iv) inhibition of lipolysis, v) inhibition of gluconeogenesis in liver, vi) an increased glycogen synthesis in liver and muscle tissue, vii) stimulation of amino acid uptake, viii) an increased protein synthesis in muscles, and ix) inhibition of proteolysis.

The blood glucose elevation occurring shortly after an intake of a meal rich in carbohydrates stimulates the secretion of insulin and concomitantly suppresses the secretion of glucagon. The combined effect thereof is a promotion of uptake of glucose into the liver, stimulation of glycogen synthesis and suppression of glycogen breakdown. When blood glucose levels subsequently begin to fall, the above events are reversed. A decreased secretion of insulin and an increased secretion of glucagon lead to the breakdown of glycogen in the liver and triglycerides in adipocytes.

Triglycerides are converted into fatty acids that are used by hepatic and muscle tissues. At the same time, the decreased insulin levels serve to reduce glucose utilization by hepatic, muscle and adipose tissues.

Type 2 diabetes accounts for approximately 80-90% of all diabetes cases and is arguably the fastest growing global threat to public health. The number of people with diabetes rose from approximately 108 million in 1980 to approximately 425 million in 2017 and the global prevalence of diabetes among adults over 18 years of age rose from approximately 4.7% in 1980 to approximately 8.8% in 2017.

Left unchecked, the current trend has been estimated by the International Diabetes Federation (IDF) to result in approximately 625 million sufferers from diabetes world wide by the year 2045. Disturbingly, some 50% of all individuals with diabetes are undiagnosed, especially in developing countries.

Various clinical studies have implicated obesity as a risk factor for type 2 diabetes although the underlying mechanism for its role in the pathogenesis of the disease is still unclear. Obesity amongst people, who subsequently develop type 2 diabetes, as well as those with existing type 2 diabetes, is associated with an increased hepatic output and reduced glucose utilization by peripheral tissues, such as e.g. muscles.

Fatty acid metabolism is increased in both obesity and in type 2 diabetes, and this may affect glucose utilization by interfering with the actions of insulin.

The development of type 2 diabetes is progressive and likely to be a culmination of pathophysiological changes occurring over many years. In most cases, the subject is unaware of the disease process, particularly in the early stages. The first stage of the disease is thought to be initiated due to a resistance to insulin. Insulin resistance is strongly associated with, and probably a major contributor to, the disease eventually entering the diabetic state. The insulin resistance stage is characterized by reduced sensitivity to insulin, as the cells normally stimulated by insulin are less sensitive to the hormone. The next stage of the disease is that of impaired glucose tolerance (IGT). IGT follows from a continued increase in insulin resistance, i.e. a continued decrease in insulin sensitivity. Impaired glucose tolerance is formally defined as a fasting venous plasma glucose concentration < 7.0 mmol/ (126 mg/dl) and a two-hour venous plasma value after 75 gram oral glucose intake > 7.8 (> 140) and < 11 ,1 mmol/l (< 200 mg/dl). When glucose concentration > 11.1 mmol/l, i.e. a level indicative of type 2 diabetes, the risk of developing specific diabetic complications is greatly enhanced. However, IGT and type 2 diabetes are both associated with a 2-4 fold increase in the burden of cardiovascular diseases. The World Health Organization (WHO) glycaemic criteria have been applied in a number of studies of IGT. These studies have determined the rate of progression to type 2 diabetes - 5 to 10 years after detection of IGT - at between 19% and 61%.

The final phase of type 2 diabetes development is characterized by insulin secretory failure (ISF). In this stage of type 2 diabetes, the insulin secretory response is inadequate. It is believed that this results from impairment of the pancreatic b-cell functions and/or the inability of b-cells to secrete sufficient amounts of insulin to compensate for the increased insulin resistance. It is to be understood that all of the above-mentioned phases in the development of type 2 diabetes leading to overt diabetes will be considered as being comprised by the term type 2 diabetes as used herein. Accordingly, a diagnosis of impaired glucose tolerance and/or reduced insulin sensitivity will be understood to relate to an individual also diagnosed as suffering from type 2 diabetes, or a condition, including any precondition, leading to type 2 diabetes.

The progression from normal glucose tolerance (NGT) to impaired glucose tolerance (IGT) is characterized by i) an increasing insulin resistance or a decreasing insulin sensitivity and ii) gradual increases in both fasting and glucose-stimulated plasma insulin levels. As IGT gradually progresses to a mild fasting hyperglycemia, there may be a further small increase in insulin resistance in both fasting and glucose-stimulated insulin release. However, in this situation, further increases in the rate of insulin secretion are no longer sustained and overt fasting hyperglycemia and increased post load glucose intolerance start to emerge. Blood glucose may be monitored directly or by measuring e.g.

HbAi_c/glycohaemoglobin in blood. The measurement of HbAi_c/glycohaemoglobin in blood has become the gold standard for the long-term control of the Glycaemic state of diabetic patients as presented in the DCCT (The Diabetes Control and Complications Trial) and UKPDS (Association of glycaemia with macrovascular and microvascular complications of type 2 diabetes) studies (Jeppsson JO, Kobold U, Barr J, Finke A, Hoelzel W, Hoshino T, Miedema K, Mosca A, Mauri P, Paroni R, Thienpont L, Umemoto M, Weykamp C; Approved IFCC reference method for the measurement of HbAi_c in human blood. Clin Chem Lab Med 2002; 40 (1):78-89; Gaborit B, Nicolay A, Valero R, Begu A, Badens C, Bellanne-Chantelot C, Portugal H, Vialettes B, Comparison of performances of various HbA1c methods in Haemoglobin Camperdown variant detection: consequences in diabetes management, Clin Chim Acta. 2009 May; 403( 1 -2) : 262-3) .

Red blood cells contain haemoglobin. Adult haemoglobin (HbA) is a protein, containing two a and two b chains. The glycosylation takes place at the N-terminal group of the b-chain at the free amino group of the amino acid valin. Glycosylated haemoglobin is called HbAi, the unglycosylated fraction which is the main part with about 90% of HbA, is called HbAo. Binding of glucose (in the following it is referred to as sugar) to haemoglobin produces 'glycosylated haemoglobin', called haemoglobin A1C or HbAic. The more sugar is in the blood, the more haemoglobin A1C or HbAic will be present in the blood.

The HbAi_c test takes advantage of the lifecycle of red blood cells. Although constantly replaced, individual red cells live for 8 -12 weeks before they are replaced. Measuring the HbAic can tell you how high your blood sugar has been on average over the last 8- 12 weeks. A normal non-diabetic HbAic is 3.5-5.5% (this varies between hospitals). In diabetes 4-6% is acceptable, whereas if the HbAic level is consistently (i.e. in several tests) ³6.5% it is possible to relate this to a diagnosis of diabetes (International Expert Committee report on the role of the A1C assay in the diagnosis of diabetes. Diabetes Care. 2009;32:1327-1334 and Use of Glycated Haemoglobin (HbAic) in the Diagnosis of Diabetes Mellitus: Abbreviated Report of a WHO Consultation, 2011).

The hemoglobin A1c test (also called HbAi_c) is a simple lab test that shows the average amount of sugar that has been in the blood of a person over the last 3 months. The hemoglobin A1c test shows whether a person's blood sugar is close to normal or is too high. It is the best test for a health care provider to tell if a person's blood sugar is under control.

Sugar in the bloodstream can become attached to the hemoglobin (the part of the cell that carries oxygen) in red blood cells. This process is called glycosylation. Once the sugar is attached, it stays there for the life of the red blood cell, which is about 120 days. The higher the level of blood sugar, the more sugar attaches to red blood cells.

The hemoglobin A1c test measures the amount of sugar sticking to the hemoglobin in the red blood cells. Results are given in per cent. Because of its “memory effect” this measure is more reliable than even repeated individual measurements of blood glucose.

The findings of a major diabetes study, the Diabetes Control and Complications Trial (DCCT) and the related Epidemiology of Diabetes Interventions and Complications (EDIC) observational follow-up (David M. Nathan and for the DCCT/EDIC Research Group, The Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications Study at 30 Years: Overview, Diabetes Care. 2014 Jan; 37(1): 9-16), have shown just how important the hemoglobin A1c test is. The study showed that lowering the hemoglobin A1c can delay or prevent the development of serious eye, kidney, and nerve disease in people with diabetes. The study also showed that lowering hemoglobin A1c levels by any amount improves a person's chances of staying healthy.

The hemoglobin A1c goal for people with diabetes is less than 7 per cent. The DCCT/EDIC findings showed that people with diabetes who keep their hemoglobin A1c levels close to 7 per cent have a much better chance of delaying or preventing diabetes problems that affect the eyes, kidneys, and nerves than people with hemoglobin A1c levels of 8 per cent or higher. A change in treatment is needed if the hemoglobin A1c level is over 8 per cent. However, if people with diabetes can lower their hemoglobin A1c by any degree, they will improve their chance of improving their conditions.

People with high daily blood sugar readings most of the time will usually have a high hemoglobin A1c test result. To maintain a hemoglobin A1c level less than 7 per cent means that the blood sugar should rarely go above 150 mg/dl on any self-monitoring blood glucose test performed before meals during the previous 3 months. As already mentioned, many cardiovascular risk factors are abnormally high in individuals suffering from type 2 diabetes and these individuals are consequently at an increased risk of developing various arteriosclerotic vascular diseases. There is at present no simple cure or medical treatment for microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system and no effective preventive therapy for treating diabetics suffering from the symptoms of cardiovascular diseases exists.

The effect of the development of microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system in diabetes is very clear. The proportion of total deaths from coronary heart disease (CHD) in diabetes has progressively increased and is now reported to cause almost 75 percent of all deaths among type 2 diabetics. However, even though the association between diabetes, insulin resistance, blood lipids, hypertension and premature arteriosclerosis have long been recognized, the complex mechanisms responsible for this association are still as vague and evasive as they were more than 50 years ago.

For type 2 diabetes there has actually not occurred a medical breakthrough since i) insulin was discovered in 1922 and since ii) sulphonylureas, biguanides, and a-glucosidase inhibitors were developed in 1954, 1957 and 1986, respectively.

Obesity and insufficient physical exercise have been suggested to be major contributors to type 2 diabetes, and coronary heart diseases and heart infarction are among the most common cardiovascular conditions diagnosed in diabetic subjects. It has been estimated that two out of every three diabetics contract a cardiovascular disease.

An increased serum level of triglycerides is now regarded as a prominent risk factor for the development of a cardiovascular diseases, also in diabetic subjects.

Pulmonary diseases are diseases generally affecting the lungs. The airways and the lungs are subject to many disease-causing and/or disease-stimulating factors such as e.g. inhaled pathogens like bacteria and viruses, allergens and toxic substances such as cigarette smoke or air pollutants. Such factors generate disorders with symptoms like e.g. difficulty in breathing, chest pains, coughing, and wheezing. The airways of the human and animal body consist of a series of tubes and passages that include the throat, the larynx and the trachea. In the chest cavity the trachea divides into the right and left bronchi, or bronchial tubes, that enter the lungs. The branches of the bronchi subsequently become more narrow and form tubes, the bronchioles, that divide into even more narrow tubes, the alveolar ducts. The end of each alveolar duct forms a cluster of thinly walled sacs termed the alveoli.

Several terms have been used to describe a group of conditions now generally recognized as leading to a limitation or obstruction of the flow of air in the airways and in the lungs. Obstructive pulmonary disease (OPD) and chronic obstructive pulmonary disease (COPD) are clinical terms describing diseases characterized by an obstruction or limitation of airflow during expiration. For COPD the obstruction or limitation is persistent. The terms represent a clinical rather than a pathological diagnosis and relate to diseases such as e.g. inflammation of the airways, asthma, bronchitis, and small airways diseases. However, the nomenclature in the field of obstructive pulmonary diseases is complex and sometimes confusing in spite of many attempts to define conditions such as asthma and bronchitis.

It is widely recognized that COPD is not a disease entity, but rather a complex of conditions characterized by airflow limitation or obstruction. The limitation or obstruction may be variable over short periods of time and reversible, even though an underlying irreversible trait may persist. Unless treated, the disease is likely to progress and lead to a seriously reduced airflow limitation. This reduction is usually, but not always, persistent and typically shows a more rapid progressive deterioration with age than normal. Clinical studies of acute exacerbations of obstructive pulmonary diseases are difficult because of i) the heterogeneous nature of COPD, ii) diffuse symptoms that can vary spontaneously, and iii) difficulties in defining a clinical response both in the short term and in the long run. Also, the role of e.g. bacterial infections and the subsequent use of antibiotics in connection with pulmonary diseases is controversial, and much evidence shows that although bacterial infections have a significant role in acute exacerbation, the role of said infections in the progression of obstructive pulmonary diseases is less certain.

Accordingly, any of the above-mentioned conditions - whether transient or chronic - may result in an airflow limitation or obstruction and may therefore be potentially associated with obstructive pulmonary diseases. The conditions may, however, also be present anatomically without generating an impairment of pulmonary function that is sufficient to qualify for the definition OPD or COPD.

An obstruction of the airways is measured by FEVi as forced expiratory volume in the first second of expiration. Lung function measured as the FEVi increases into young adulthood and then it starts to decrease. In normal non-smokers, the rate of decline in FEVi is about 20 ml per year, i.e. about 1 liter over a 50-year period. A much more rapid decline is observed in smokers. On average, the decline is twice that of normal non-smokers. However, in about 15% of all smokers, lung function declines at a rate much more rapid than the decline observed in the average smoker. Consequently, airways diseases are strongly influenced by individual rates of decline in FEVi.

Asthma has traditionally been regarded as a respiratory disease of acute airway obstruction, and research as well as therapeutic attention has focused principally on the mechanisms leading to acute bronchospasm. One of the conventional therapies has consisted of bronchodilators to regulate airway smooth muscle contraction.

However, current state of the art asthma therapy does have side effects, mostly due to undesirable effects from the inhalation steroids used.

A wide range of pharmaceuticals have been developed by the pharmaceutical industry and evaluated in clinical trials. Although being capable of inhibiting mast cell-mediated acute allergic bronchoconstriction, none of these pharmaceuticals are suitable for use in a prophylactical treatment or maintenance treatment of asthma. Medicaments such as P2 agonists have been introduced in order to treat airways diseases and in particular asthma. P2 agonists inhibit the release of histamine into the circulation of asthmatics undergoing an allergen provocation. This pharmacological property may contribute to the well-recognized ability of P2 agonists to inhibit allergen-induced bronchoconstriction. However, while P2 agonists are exceptional mast cell stabilizing agents, sole therapy with these agents may actually enhance hyperresponsiveness of airways to exogenous stimuli such as inhaled histamine, most likely due to a minimal effect on airway inflammation.

Widespread use of P2 agonists have led to a criticism based on a hypothesis involving the so-called "asthma paradox". According to the hypothesis, P2 agonists have undesirable effects on the normal role of mast cell degranulation as an endogenous anti-inflammatory mechanism to prevent antigens from entering the lower airways and limit the extent of the subsequent repair process.

Both Rietjens et al (Ivonne M C M Rietjens, Jochem Louisse and Karsten Beekmann,

The potential health effects of dietary phytoestrogens, Br J Pharmacol. 2017 Jun;

174(11): 1263-1280) and Adlercreutz (Finnish Medical Society, Ann. Med. 29, 95-120 (1997)) have reviewed the phytoestrogen classes of lignans and isoflavones and has described their influences on a range of cellular activities and metabolic events. Soy intake is reported to influence the development of menopausal symptoms, cardiovascular disease and brain function disorders but no exact mechanisms have been established. Both authors emphasize that phytoestrogens, particularly in association with soy intake, seem to have a great potential for preventing cardiovascular diseases, but that an established beneficial effect of soy and isoflavonoids will have to await further studies. Isoflavones may form a group of substances with a great potential in preventive medicine. Also the authors make no definite recommendations as to the amounts needed for disease prevention.

Xiao (Chao Wu Xiao, Health Effects of Soy Protein and Isoflavones in Humans, The Journal of Nutrition, Volume 138, Issue 6, June 2008, Pages 1244S-1249S) and Messina (Soy and Health Update: Evaluation of the Clinical and Epidemiologic Literature, Nutrients. 2016 Dec; 8(12): 754) analysed a number of clinical trials and concluded that the consumption of soy protein significantly decreases serum levels of total cholesterol, LDL-cholesterol and triglycerides in hyperlipidemic subjects and reported that isoflavones may be of great potential benefit to human health maintenance and that isoflavones may be health-protective in amounts potentially available from a human diet containing daily soy foods. It is emphasized, however, that the relation between concentration of isoflavones in human plasma and the biological effects of the isoflavones remains controversial. Long-term human feeding trials of purified isoflavones and foods containing isoflavones to examine health-related outcomes may be warranted.

WO 97/31546 discloses data from total replacement programs (for 6 weeks) in weight reduction studies conducted at Karolinska Hospital in Sweden. It is shown that products comprising isolated soy protein and soybean cotyledon fibers lower serum triglyceride levels by a maximum of 44 percent and cholesterol levels by a maximum of 27 percent for a patient population with a mean initial cholesterol content of 5.6 mmol/l. A mean value of 6.25 mmol/l was determined for all patients having serum cholesterol levels above 6 mmol/l, and for this group of patients a reduction in serum cholesterol levels of 33 percent was observed. Since the reported data were part of a weight reduction program, a dietary effect and/or an effect related to a weight loss would have contributed to the observed reductions in cholesterol and/or triglycerides. No reference is made to a treatment of diabetes by using a composition comprising a combination of soy protein, a high content of a phytoestrogen compound, and a phospholipid source comprising high fixed levels of phosphatidyl choline. No reference is made to a treatment of a pulmonary disease by using a composition comprising a combination of soy protein, a high content of a phytoestrogen compound, and a phospholipid source comprising high fixed levels of phosphatidyl choline. No reference is made to a composition comprising a combination of soy protein, a high content of a phytoestrogen compound, and soy lecithin comprising high fixed levels of phosphatidyl choline or a composition comprising a combination of soy protein, a high content of a phytoestrogen compound, lecithin comprising high fixed levels of phosphatidyl choline and dietary fibers.

W00030663 ( PCT /I B99/01992) , W00030664 (PCT/IB99/01997) and W00030665 (PCT/IB99/01998) all relate to compositions comprising (a) soy protein, (b) a phytoestrogen compound, and (c) dietary fibres. The soy protein (a) is present in an amount of at least 45 weight percent of the total protein content of the composition, said total protein content providing at least 15 percent of the total energy content of the composition. The phytoestrogen compound (b) is preferably a naturally occurring isoflavone and is present in an amount of more than 0.10 weight percent of the soy protein, and the dietary fibres (c) are preferably soybean fibres and are present in an amount of more than 4 weight percent of the total weight of the nutritional composition on a dry basis. The composition of W00030663 is claimed to be useful for treating type 2 diabetes and cardiovascular diseases in a diabetic subject. The composition of W00030664 is claimed to be useful for treating pulmonary diseases. The composition W00030665 is claimed to be useful in lowering serum cholesterol and LDL-cholesterol and serum triglyceride levels and for increasing the HDL/LDL-cholesterol ratio in serum of a subject suffering from arteriosclerosis and related cardiovascular diseases. Compositions further comprising an additional fat source, i.e. soy lecithin, are described. One major source of dietary phospholipids is lecithin, and the major part of the phospholipids of lecithin are phosphoglycerides. Lecithin was first isolated from egg yoke in 1850 by Maurice Bobley. Since that time, it has been shown to be present in many foods. Soybeans and other legumes, grains, wheat germ, brewers yeast, and fish, as well as egg yokes are all good sources of lecithin.

In 1958 L. M. Morrison was the first to report the finding that lecithin could be used to lower cholesterol levels. L. M. Morrison speculated that instead of "blocking" absorption of cholesterol in the digestive tract as other cholesterol reducing agents did, lecithin enhanced the metabolism of cholesterol in the digestive system and aided in its transport through the circulatory system. Researchers have since demonstrated that atherosclerosis (blockage of the arteries) can be induced in the laboratory by either increasing the cholesterol introduced into the body or by decreasing lecithin intake.

Mourad et al (Amouni Mohamed Mourad, Eder de Carvalho Pincinato, Priscila Gava Mazzola, Maricene Sabha, and Patricia Moriel, Influence of Soy Lecithin Administration on Hypercholesterolemia, Cholesterol Volume 2010, Article ID 824813,

4 pages) have found that soy lecithin-rich diets can be used as an adjunct in the treatment of hypercholesterolemia, and by now it is generally accepted that lecithin from a vegetable source (soybeans) is more effective than lecithin from an animal source (eggs) in accelerating re-absorption of cholesterol back into the blood stream that has adhered to the walls of blood vessels and caused blockage. This difference might be attributed to the fact that lecithin from animal sources contains high amounts of saturated fatty acids, while lecithin from vegetable sources are about 80% unsaturated fatty acids.

Besides being an important factor in generally controlling cholesterol levels and aiding coronary health, lecithin, and especially the phosphoglyceride phosphatidyl choline, has been speculated to be involved in numerous metabolic processes associated with e.g. arterioschlerosis, type 2 diabetes, obesity, etc. Accordingly, versatile effects of phosphatidyl choline consumption, i.e. modulation of lipid exchange between cell membranes and the plasma compartment, modulation of gallbladder bile content, inhibition of intestinal absorption of cholesterol, protection against LDL oxidation, decrease in LDL cholesterol, decrease in serum cholesterol, increase in HDL- cholesterol and plasma apolipoprotein A-l, decrease in LDL cholesterol/HDL cholesterol ratio. However, up till now several authors have questioned whether any independent effects on serum lipoprotein could be attributed to phosphatidyl choline.

W003004039 relates to soy protein, phytoestrogens, phospholipids, and dietary fibers and compositions thereof suitable for preventing, treating and/or alleviating cardiovascular diseases such as hypercholesterolemia, hypertriglyceridemia, hyperlipidemia, arteriosclerosis, hypertension and related cardiovascular diseases, for preventing and/or treating type 2 diabetes and/or the metabolic syndrome, and for preventing, treating and/or alleviating pulmonary diseases. The compositions described comprise (a) soy protein making up at least 45 weight percent of the total protein content of the composition, said total protein content providing at least 15 percent of the total energy content of the composition, (b) phytoestrogens in an amount of more than 0.10 weight percent of the soy protein content of the composition, and (c) a phospholipid source providing at least 15 percent of the total energy content of the composition, and (d) phosphatidyl choline in an amount of more than 10 weight percent of the phospholipid source of the composition.

CN 101 248 881 describes a food supplement composition comprising isolated soy protein (6.25-12.50 weight percent) and soy isoflavones (1.25-3-12 weight percent).

The composition is used for treating menopausal symptoms and isoflavones are described as providing anti-tumor activity and an effect on cardiovascular protection.

Cvejic et al (Jelena Cvejic, Dorde Malencic, Vesna Tepavcevic, Mihalj Posa and Jegor Miladinovic, Determination of Phytoestrogen Composition in Soybean Cultivars in Serbia, Natural Product Communications vol. 4, no. 8, August 2009, p 1069-1074) describes the total phytoestrogen concentration in different soy bean varieties from Serbia as being between 2.24 and 3.79 mg/g dry bean weight. The phytoestrogens were isolated from the beans by grounding in a coffee mill, followed by defatting with n-hexane and extraction with methanol-water (8:2 v/v) and measured as daidzein and its derivatives, genistein and its derivatives, and glycitein and its derivatives, respectively. It is set out that in the analyzed soybean varieties, the isoflavones were mostly present in the form of glycosides, and that the average content of isoflavones in the free aglycon forms was only 0.92% of the total isoflavone content. It is, furthermore, set out that the phytoestrogen contents measured in relation to the Serbian varieties in question are considerably higher than similar values previously measured for other (US) soybean varieties, and that the phytoestrogen content may, thus, vary considerably between different varieties. No information as to the average protein content of the soybean varieties analyzed is provided.

None of the above-mentioned documents discloses compositions having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising (a) a soy protein source, which provides for at least 12.5 g of soy protein and at least 50 weight percent of the total protein content of the composition, and (b) a source of naturally occurring phytoestrogens in the form of a soy protein source with a high concentration of naturally occurring isoflavones and/or a soy germ extract providing naturally occurring phytoestrogen compounds in an amount of minimum 50 mg and sufficient to ensure that the total content of naturally occurring isoflavones of the composition amounts to at least 0.34 weight percent but less than 2.50 weight percent of the soy protein content of the composition, for use in preventing, treating and/or alleviating; microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system; menopausal symptoms or premenstrual syndrome; high blood pressure, elevated blood lipid and blood sugar levels; hormonal prostate, breast or colon cancers; inflammation of joints and tendons; pulmonary diseases such as e.g. airway inflammation, asthma, bronchitis and small airways diseases; or for use in improving physical performance and/or athletic performance or vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles.

SUMMARY OF THE INVENTION

The present invention provides a nutritional composition comprising a soy protein source, and a soy germ source, providing a high, fixed amount of a phytoestrogen compounds in the form of naturally occurring isoflavones. More particularly the present invention provides a nutritional composition of soybean extractable ingredients having a high, fixed amount of naturally occurring isoflavones.

The present invention provides the use of composition comprising a) soy protein, preferably isolated soy protein, b) a high content of naturally occurring isoflavones such as found in soy germ, and optionally (c) a phospholipid source, more preferably lecithin, and even more preferably soy lecithin and preferably having a high fixed level of phosphatidyl choline, and optionally (d) dietary fibers, preferably soybean fibers, more preferably soybean fibers manufactured from the cotyledon of soybeans hereinafter referred to as soy cotyledon fibers. The compositions of the present invention are useful in preventing, treating and/or alleviating including prophylactically treating microvascular or lipoprotein related diseases of the heart; vascular and/or central nervous system, menopausal symptoms or premenstrual syndrome; high blood pressure, elevated blood lipid and blood sugar levels; hormonal prostate, breast or colon cancers; inflammation of e.g. joints and tendons; pulmonary diseases such as e.g. airway inflammation, asthma, bronchitis and small airways diseases. In addition, the compositions of the present invention are useful in improving physical performance and/or athletic performance or vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles.

It is one objective of the present invention to significantly lower levels of total serum cholesterol and LDL-cholesterol and triglycerides in a mildly hypercholesterolemic subject. It is another objective of the present invention to significantly lower serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides in a subject suffering from hypercholesterolemia and/or hyperlipidemia. It is another objective of the present invention to render the arterial wall more resistant to the accumulation of lipoproteins. It is a further objective of the present invention to provide a composition effective in preventing, treating, prophylactically treating and/or alleviating an arteriosclerotic condition by reducing the influx of cholesterol and/or triglycerides through the endothelium into the arterial wall, and/or by causing the dilation of blood vessels. Yet another objective of the present invention is to reduce lipid plaque formation and inflammation in blood vessels.

The present invention is also useful in the prevention and/or treatment of type 2 diabetes and/or a cardiovascular disease in diabetic subjects. Accordingly, it is an objective of the present invention to effectively lower serum levels of both glucose and cholesterol and/or triglycerides. It is to be understood that diabetic subjects according to the present invention have a fasting plasma glucose > 7.0 mmol/l.

A composition according to the present invention represents a new approach to treatment of type 2 diabetes and is believed to be capable of preventing, treating and/or alleviating an arteriosclerotic condition by reducing the influx of cholesterol and/or triglycerides through the endothelium into the arterial wall of a diabetic subject suffering from a cardiovascular disease, and/or by causing the dilation of blood vessels. No other known compositions are effective in reducing the influx of lipids such as e.g. cholesterol and/or triglycerides into the arterial wall, and to dilate blood vessels.

The present invention is also useful in the prevention and/or effective treatment of pulmonary diseases such as e.g. airway inflammation, asthma, bronchitis and small airways diseases, in particular asthma including chronic asthma such as e.g. asthma characterized by a chronic inflammatory condition. The present invention is believed to be capable of increasing FEVi of a subject, measured by forced expiratory volume in the first second of expiration, as well as being capable of treating, alleviating and/or eliminating in particular i) inflammation of the airways, ii) mucus hypersecretion, and iii) bronchoconstriction.

In addition, the present invention is useful in improving physical performance and/or athletic performance through vasodilation, and/or vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles.

A composition according to the present invention may be comprised of a micronutrient as defined herein below.

Phytoestrogen compounds are naturally occurring plant hormones showing a structural similarity to 17p-estradiol. Phytoestrogens consist of a number of classes including isoflavones, coumestans, lignans and resorcylic acid lactones. The class of isoflavones consists of among others genistein, daidzein, equol, glycitein, biochanin A, formononetin, and O-desmethylangolesin. The isoflavones genistein and daidzein are found almost uniquely in soybeans. When present in the plant the isoflavones are mainly in a glucoside form, i.e. attached to a sugar molecule. Isoflavones in this glucoside form can be deconjugated to yield isoflavones in a so-called aglycone form, which is the biologically more active form of isoflavones and which is absorbed faster and to a greater extent in the human gut than isoflavones in the glucoside form. In vitro studies have examined the relative estrogenic effect exerted by various phytoestrogens including isoflavones. The resulting potencies as compared to estradiol (having a relative potency of 100), have been reported by Knight (Maturitas 22, 167-175 (1995)) for among others genistein (0.084) and daidzein (0.013).

However, the results also showed that the estrogen receptor complexes formed by estradiol and isoflavones such as genistein and daidzein are functionally equivalent.

The comparative dissociation constant of genistein for the estrogen receptor, as determined in competitive binding assays, was found to be from 100 to 10.000 times higher than that of estradiol.

The term "naturally occurring" substance as used in the present specification refers to a substance originally isolated from a natural source, such as a plant, for example a soy plant, or modified forms of such a substance. The naturally occurring substance for use in a composition according to the present invention may be included in a composition according to the present invention as part of the natural source or in any type of extract, isolate or the like thereof.

Soy proteins are involved in a reduction of cholesterol and triglyceride levels, dilation of blood vessels by releasing nitric oxide (NO), they are easily digestible, and they represent an efficient sole protein source for maintaining the nitrogen balance. Soy isoflavones in high intakes further enhances these effects and provide anti inflammatory effects. Phospholipids, such as soy lecithins, especially soy phosphatidyl choline have been shown to lower total serum cholesterol levels and/or to increase serum HLDL-cholesterol levels. Dietary fibers, such as soybean fibers, especially soy cotyledon fibers have been shown to lower total serum cholesterol levels, to improve glucose tolerance, to increase insulin sensitivity, to normalize the gastrointestinal function, and to exert no influence on the absorption of essential minerals.

Accordingly, in one aspect the present invention provides a composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising

(a) a soy protein source, selected from isolated soy protein, soy protein concentrate, or soy flour, which provides for at least 12.5 g of soy protein and at least 50 weight percent of the total protein content of the composition and

(b) at least one naturally occurring phytoestrogen compound in the form of naturally occurring isoflavones in an amount of minimum 50 mg and more than 0.34 weight percent of the soy protein content of the composition

In a more preferred aspect the present invention provides a composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising (a) a soy protein source, selected from isolated soy protein, soy protein concentrate, or soy flour, which provides for at least 12.5 g soy protein and at least 50 weight percent of the total protein content of the composition, and

(b) at least one naturally occurring phytoestrogen compound in the form of naturally occurring isoflavones in an amount of minimum 50 mg and more than 0.34 weight percent of the soy protein content of the composition, and

(c) a phospholipid source in an amount sufficient to ensure that the total content of soy phospholipids of the composition amounts to at least 15 weight percent of the soy protein content of the composition

In a presently most preferred aspect the present invention provides a composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising

(a) a soy protein source, selected from isolated soy protein, soy protein concentrate, or soy flour, which provides for at least 12.5 g soy protein and at least 50 weight percent of the total protein content of the composition, and

(b) at least one naturally occurring phytoestrogen compound in the form of naturally occurring isoflavones in an amount of minimum 50 mg and more than 0.34 weight percent of the soy protein content of the composition,

(c) a phospholipid source in an amount sufficient to ensure that the total content of soy phospholipids of the composition amounts to at least 15 weight percent of the soy protein content of the composition, and

(d) a fibre source, such as a soy fibre source, such as a soy cotyledon fibre source in an amount sufficient to ensure that the total content of fibres, such as soy fibres, such as soy cotyledon fibres of the composition amounts to at least 25 weight percent of the soy protein content of the composition,

In further preferred aspects of the present invention the above-mentioned composition may comprise a phospholipid source in an amount sufficient to ensure that the total content of soy phospholipids of the composition amounts to at least 10, 11, 12, 13 or 14 weight percent of the soy protein content of the composition, and a fibre source, such as a soy fibre source, such as a soy cotyledon fibre source in an amount sufficient to ensure that the total content of fibres, such as soy fibres, such as soy cotyledon fibres of the composition amounts to at least 20, 21, 22, 23 or 24 weight percent of the soy protein content of the composition.

Phytoestrogen compounds according to the present invention are defined as naturally occurring plant substances, which are either structurally or functionally similar to 17b- estradiol or generate estrogenic effects. Phytoestrogens consist of a number of classes including isoflavones, coumestans, lignans and resorcylic acid lactones.

Examples of isoflavones according to the present invention are genistein, daidzein, equol, glycitein, biochanin A, formononetin, and O-desmethylangolesin. The phytoestrogen compounds of a composition according to the present invention are preferably isoflavones, more preferably genistein, daidzein and/or glycitein, yet more preferably genistein and/or daidzein and even more preferably genistein. Genistein and daidzein are found almost uniquely in soybeans. A preferred composition according to the present invention may accordingly comprise a single isoflavone, such as genistein, daidzein or glycitein, or it may comprise at least one isoflavone selected from the group comprising at least genistein, daidzein and glycitein. Furthermore, a preferred composition according to the present invention comprise isoflavones being naturally part of the soy protein and/or soy germ sources employed.

Phospholipid sources according to the present invention are defined as fat substances comprising at least about 5% phosphatidyl choline. However, phospholipid sources according to the present invention may contain as much as 100% phosphatidyl choline. Furthermore, phospholipid sources according to the present invention will preferably comprise polyunsaturated fatty acids and monounsaturated fatty acids and optionally also saturated fatty acids. The phospholipid sources will preferably comprise polyunsaturated fatty acids and monounsaturated fatty acids and optionally also saturated fatty acids. The amount of polyunsaturated fatty acids and monounsaturated fatty acids, including the essential fatty acids, may range from 35 to 50, preferably 38 to 44, weight percent of the total amount of the phospholipid source source. The essential fatty acids are also called omega-6 and omega-3 fatty acids and include linolic acid and/or linolenic acid (a-linolenic acid). The amount of saturated fatty acids may be from 20 to 30 weight percent, preferably 22 to 26 weight percent, of the total amount of the phospholipid source. Lecithins and soy lecithins having a high content of a-linolenic acid are particularly preferred phospholipid sources according to the present invention.

A composition according to the present invention is capable of preventing, treating, prophylactically treating and/or alleviating an arteriosclerotic condition by reducing the accumulation of cholesterol in the arterial wall and/or causing the dilation of blood vessels and by providing anti-inflammatory effects. These inhibitory effects may be mediated by the binding of naturally occurring isoflavones and/or soy peptides to an estrogen receptor or estrogen-like receptor present in the endothelium of an artery and/or through the action of the 1-alkyl-2-acetyl analog of phosphatidyl choline. The soy peptides are preferably provided by partial hydrolysis of soy protein.

Plasma cholesterol and triglyceride levels are usually increased in individuals treated for a cardiovascular disease and plasma triglyceride and lipoprotein levels are usually increased in individuals treated for type 2 diabetes and/or the metabolic syndrome. These increased levels, unless reduced by treatment, are likely to promote atherosclerosis and/or coronary heart disease (CHD). Beta-2-adrenergic receptors are present on many different types of cells including i) cells of the arterial wall ii) cells of the airways and iii) fat cells. Beta-2-adrenergic receptors are involved in the regulation of triglyceride synthesis in fat cells and according to one presently preferred hypothesis, binding of soy peptides and/or a phytoestrogen compound such as e.g. a naturally occurring isoflavone to a beta-2-adrenergic receptor present on a fat cell or in an arterial wall is effective in reducing e.g. the synthesis of triglycerides in fat cells and/or the release of triglycerides into the blood stream and/or reducing the influx of cholesterol and/or triglycerides into the arterial wall. The soy peptides are e.g. obtainable by partial hydrolysis of soy protein.

Several mechanisms for the association between elevated homocysteine and vascular diseases have been proposed, including effect on endothelial function, vascular smooth muscle cells, LDL-C action, coagulation pathways, and oxidative status. In addition to the effect of homocysteine on CVD, some data indicate elevated homocysteine levels are related to poor cognitive function and cancer. Three nutrients- -folate, vitamin B6 and vitamin B12-have been shown to influence homocysteine levels. However, an additional compound, betaine, is required for the conversion of homocysteine to methionine. Betaine, which acts as a methyl donor, is synthesized endogenously from choline through the action of choline dehydrogenase and betaine aldehyde dehydrogenase. In children with cystathione beta-synthase deficiency, which results in the accumulation of homocysteine due to the inability to convert homocysteine to cystathionine, betaine administration has been shown to lower homocysteine levels significantly. In other cases, homocysteine lowering by folate requires the addition of choline or betaine. Thus soy phospholipids, e.g. phosphatidyl choline may act as a direct controlling factor in lowering homocysteine levels and thereby reduce and/or eliminate one of the risk factors associated with cardiovascular diseases.

As an integral part of cell membranes, phospholipids are directly involved in cell signaling. As a constituent of cell membranes, phospholipids consist of a variety of molecular species, including ester-, ether-, and vinyl-linked forms. In response to cell stimuli, phospholipids are broken down by specific phospholipases, resulting in a number of hydrolysis products that have the potential to act as second messengers and thus markedly influence cellular processes, including cholesterol metabolism.

According to a preferred hypothesis, a composition according to the present invention will reduce and/or eliminate one or more of the risk factors for cardiovascular diseases. Accordingly, a composition according to the present invention may be effective in preventing, treating, prophylactically treating and/or alleviating conditions such as e.g. microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system, menopausal symptoms or premenstrual syndrome, high blood pressure, elevated blood lipid and blood sugar levels, hormonal prostate, breast or colon cancers, inflammation of e.g. joints and tendons, pulmonary diseases such as e.g. airway inflammation, asthma, bronchitis and small airways diseases. A composition according to the present invention may also be capable of reducing, preventing and/or eliminating fatty streak formation and/or fibrous plaque development and/or effective in mediating a regression of one or both of said arteriosclerotic conditions. A composition according to the present invention may also be capable of improving the physical performance and/or athletic performance as measured in watts of a human or animal subject, or of improving vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles..

A composition according to the present invention may be effective in preventing and/or treating type 2 diabetes and/or the metabolic syndrome and/or reducing and/or eliminating one or more of the risk factors for cardiovascular diseases associated with diabetes and/or the metabolic syndrome. A composition according to the present invention may also be capable of reducing, preventing and/or eliminating fatty streak formation and/or fibrous plaque development and/or effective in mediating a regression of one or both of said arteriosclerotic conditions in diabetic individuals.

According to a preferred hypothesis, a composition according to the present invention will be effective in lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or in increasing the serum HDL/LDL-cholesterol ratio and/or increasing serum levels of high-density lipoproteins (HDL) and/or in generating a decrease in serum levels of low-density lipoproteins (LDL). It is desirable to achieve an elevated serum HDL/LDL-cholesterol ratio since this may result in an increased reverse cholesterol transport and a subsequent excretion.

Also, it is believed that a composition according to the present invention will affect ApoB lipoprotein metabolism including the metabolism of a recently discovered class of ApoB comprising lipoprotein particles called small, dense LDL particles. The LDL class of lipoproteins is in fact composed of several components with distinct properties. The basis for this heterogeneity and the consequences for disease are at present not thoroughly understood. An increased level of small, dense LDL particles is one of the most common dyslipoproteinemias associated with coronary artery disease, and serum levels of ApoB are often disproportionately elevated compared with LDL- cholesterol in dyslipoproteinemic patients.

Heterogeneity within lipoprotein classes may be the result of a differing lipid content, a different apoprotein composition, an altered protein conformation or an as yet unidentified structural variation. Subjects with a preponderance of small, dense LDL have an increased risk of suffering a myocardial infarction independent of the total concentration of serum LDL. Accordingly, a composition according to the present invention may be effective in lowering elevated levels of small, dense LDL.

Hypertriglyceridemia in non-diabetic and diabetic subjects alike is associated with an increase in the clotting activities of thrombogenic factors such as e.g. factor VII and/or factor X and/or factor XII and an increase in the level of the inhibitor of tissue plasminogen activator, PAI-1. The increased inhibitor concentration results in a decreased level of plasminogen synthesis and thus a decreased level of plasminogen stimulated clot lysis. These changes in clotting activities no doubt contribute to the procoagulant state, which is also observed in diabetes. Accordingly, the present invention provides a composition, which may be effective in normalizing levels of homocysteine and/or the clotting activities of at least one thrombogenic factor such as e.g. factor VII and/or factor X and/or factor XII by e.g. decreasing the increased activity thereof, which is also observed in a subject diagnosed as having type 2 diabetes or diagnosed as having an impaired glucose tolerance or a decreased insulin sensitivity.

Also, a composition according to the present invention may be effective in promoting a decrease in the level of the inhibitor of tissue plasminogen activator, PAI-1, which in turn leads to an increased plasminogen stimulated clot lysis. A composition according to the present invention may also be effective in reducing an increased platelet aggregatability and/or mediating directly or indirectly a reduction in the increased level of lipoprotein (a) associated with a procoagulant state in an arteriosclerotic condition and/or a diabetic condition.

Accordingly, in one embodiment the present invention provides a composition effective in reducing and/or eliminating risk factors for coronary heart disease (CHD) in obese subjects and in obese subjects suffering from a diabetic condition and/or the metabolic syndrome.

A composition according to the present invention may also be effective in treating dyslipidemia such as e.g. hypertriglyceridemia and/or hypercholesterolemia in connection with increased serum levels of VLDL, decreased and altered serum levels of HDL and increased serum levels of small dense LDL, and hypertension, all of which are risk factors for atherosclerosis. Accordingly, in one embodiment, a composition according to the present invention may be capable of effectively lowering and/or eliminating increased serum levels of VLDL, and/or effectively increasing decreased serum levels of HDL, and/or effectively lowering serum LDL levels including serum levels of small dense LDL. A composition according to the present invention may be capable of preventing, treating, prophylactically treating and/or alleviating hypertension.

A composition according to the present invention may also be effective in suppressing any effect that would otherwise generate an increased turnover of arterial smooth muscle cells, i.e. an enhanced arterial smooth muscle cell proliferation, and/or lead to an increased cholesterol ester accumulation in the arterial wall. In hypercholesterolemia characterized by increased levels of intracellular cholesterol resulting from e.g. increased delivery of LDL-cholesterol via the LDL receptor, a composition according to the present invention may be effective in reducing the increased activity of the LDL receptor. It is also possible that insulin and other growth factors have the potential to promote the accumulation of cholesterol intracellularly. This may in fact well occur in a diabetic subject and more generally under conditions when cells are stimulated, but cannot proliferate normally. Accordingly, a composition according to the present invention may also be capable of treating, alleviating and/or eliminating any decrease, including any insulin mediated decrease, in the HDL receptor-mediated cholesterol efflux. Accordingly, a composition according to the present invention may be capable of reducing and/or eliminating any enhanced retention of intracellular cholesterol caused by a decreasing HDL receptor-mediated cholesterol efflux.

A composition according to the present invention may be effective in reducing insulin resistance by stimulating cells or receptors located thereon that are normally stimulated by insulin, but less sensitive to the hormone in a subject diagnosed with type 2 diabetes and/or the metabolic syndrome. A composition according to the present invention may also be effective in stimulating cells comprising a beta-2- adrenergic receptor or a receptor belonging to the class of beta-2-adrenergic receptors. The final phase of type 2 diabetes development is characterized by insulin secretory failure (ISF), and in one presently preferred hypothesis, this failure is at least preventable by a composition according to the present invention effective in stimulating insulin secretion.

Hypertriglyceridemia in diabetes has been associated with a variety of changes in circulating lipoproteins, and a composition according to the present invention may be capable of preventing, treating, alleviating and/or eliminating cardiovascular risk factors such as e.g. chylomicronemia, an increased level of VLDL, an increased level of remnants (VLDL and chylomicrons), and LDL and HDL containing increased levels of triglycerides.

Lipoprotein fractions obtained from type 2 diabetic subjects tend to lose their typical sharp LDL peak and instead have a broad diffuse LDL band termed polydisperse LDL. Dissection of polydisperse LDL reveals that diabetics have an increased serum level of intermediate-density-lipoprotein (IDL), an abnormal LDL peak, and an increase in the amount of small dense LDL. While small dense LDL particles have been associated with CHD in the general population, a similar association in diabetes remains to be established. Accordingly, a composition according to the present invention may be effective in promoting a decreased serum level of intermediate density lipoprotein (IDL), a normal, sharp LDL peak, and a decreased amount of small dense LDL.

Accordingly, diabetic dyslipidemia of type 2 diabetes is generally associated with abnormalities of apolipoprotein and lipoprotein particle distributions and results in increased plasma VLDL and remnant levels, an increase in the apoE concentration in VLDL and remnants, an increase in the amount of small dense LDL, and an altered HDL particle distribution.

According to one presently preferred hypothesis, a composition according to the present invention will alleviate abnormalities associated with apolipoprotein and lipoprotein particle distribution and promote a decreased plasma VLDL and remnant level, a decrease in the apoE concentration in VLDL and remnants, a decrease in the amount of small dense LDL, and a HDL particle distribution similar to that of a comparable non-diabetic, healthy individual.

Hyperinsulinemia is also considered a risk factor for coronary heart disease (CHD) in diabetic subjects due to the association of high insulin levels with increased incidence and mortality rates of CHD. A composition according to the present invention may be effective in lowering serum insulin levels in subjects diagnosed with type 2 diabetes. Diabetic patients having increased endogenous insulin levels, i.e. subjects diagnosed with type 2 diabetes, or having increased peripheral circulating insulin levels as a result of intermittent injections of large amounts of exogenous insulin are particularly prone to hyperinsulinemia.

Hyperinsulinemia in both normal persons, persons with the metabolic syndrome and those with type 2 diabetes appears to be related to obesity. Insulin levels are very often increased in both the fasted state and after intake of a diet rich in carbohydrates in obese individuals, irrespective of whether they suffer from a diabetic condition or not. Furthermore, hyperinsulinemia appears to be directly correlated to the degree of obesity. Accordingly, hyperinsulinemia is one of the many risk factors for CHD associated with obesity, and insulin may modulate many other obesity-related risk factors. Accordingly, a composition according to the present invention may be effective in lowering insulin levels in obese subjects with diabetes or the metabolic syndrome.

In obese subjects diagnosed as diabetic, LDL particle size is independently correlated with factors such as e.g. serum triglyceride and serum insulin levels. Consequently, it is possible that the extent of adiposity and concomitant insulin resistance in hyperinsulinemic individuals is associated with the occurrence small dense LDL, independently of hypertriglyceridemia, which is another diabetic condition also putatively associated with small dense LDL formation. Accordingly, both insulin resistance and hyperinsulinemia appear to play a central role in the pathogenesis of atherosclerosis in diabetes. A composition according to the present invention may be effective in treating and/or alleviating insulin resistance and/or hyperinsulinemia.

It is very possible that type 2 diabetes is also associated with insulin resistance and hyperinsulinemia independently of an increase in abdominal lipids. Hyperinsulinemia in turn is associated with dyslipidemia, i.e. increased VLDL, decreased and altered HDL and increased small dense LDL, and with hypertension, all of which are risk factors for atherosclerosis. This array of abnormalities and disorders, or a part of thereof, is generally termed the insulin resistance syndrome, or syndrome X, or metabolic syndrome.

In one embodiment, a composition according to the present invention may be capable of effectively decreasing and/or eliminating increased serum levels of VLDL and/or LDL, and/or increasing decreased serum levels of HDL, and of decreasing and/or eliminating serum LDL levels including serum levels of small, dense LDL. A composition according to the present invention may also be capable of reducing an elevated level of small, dense LDL particles and/or reducing an elevated ratio of LDL- apoB to LDL-cholesterol and/or preventing, treating or alleviating hypertension.

Even though there is no internationally agreed definition for the metabolic syndrome, the term as used herein shall be understood to relate to the occurrence in a subject of at least two of the following: i) impaired glucose tolerance, ii) elevated blood pressure, iii) hypertriglyceridemia and low HDL-cholesterol, iv) insulin resistance, and v) obesity.

The occurrence of a condition characterized by one or more of impaired glucose tolerance, elevated blood pressure, hypertriglyceridemia and low HDL-cholesterol, insulin resistance, and obesity will depend on variables such as sex, age, body weight, physical condition and the like, and general WHO guidelines will generally be adhered to when evaluating the occurrence of any one of the above-mentioned conditions.

Hyperinsulinemia in itself may well be capable of affecting the arterial wall either directly or indirectly by promoting or facilitating the promotion of changes similar to those leading to severe atherogenesis. Insulin may well promote both arterial smooth muscle cell proliferation and cholesterol ester accumulation in the arterial wall. A composition according to the present invention may in one embodiment be effective in preventing, treating, alleviating and/or eliminating fatty streak formation, fibrous plaque development, complicated lesion formation, thrombosis, platelet aggregation and/or myocardial infarction.

Since insulin can be expected to be capable, either in combination with other compounds such as additional growth factors, or on its own, of increasing the levels of intracellular cholesterol, by e.g. increasing a delivery of LDL-cholesterol via the LDL receptor, and concomitantly therewith increase an endogenous biosynthesis of cholesterol that makes yet more cholesterol available for new membrane synthesis in the cell proliferation process, it is an object of the present invention to counteract any increased activity including any insulin stimulated increased activity of the LDL receptor.

Modifications of lipoproteins constitute another risk factor for cardiovascular disease, including cardiovascular disease in diabetes. The modification characterized by protein glycosylation is associated with e.g. arteriosclerosis and diabetes, and glycosylated lipoproteins such as e.g. LDL, IDL, VLDL and HDL can be expected to be functionally abnormal. Accordingly, the accumulation of glycosylated LDL in the plasma, including the plasma of a diabetic subject, can be perceived to enhance cholesterol ester accumulation. Also, glycosylation of HDL can be expected to impair the ability of HDL binding to the HDL receptor. This impaired binding is likely to reduce the level of intracellular cholesterol efflux. Accordingly, glycosylated HDL may well be another factor potentially contributing to the accumulation of cholesterol in the arterial cell wall.

A composition according to the present invention may be effective in preventing, treating, alleviating, reducing and/or eliminating lipoprotein glycosylation in the serum of subjects, including diabetic subjects. In addition, a composition according to the present invention may also be effective in preventing lipoprotein modification caused e.g. by oxidation, chemical modification such as chemical cross-linking, or modifications caused by an alteration in the lipid composition of the lipoprotein, such as any increase or decrease in the content of triglycerides, cholesterol esters, free cholesterol, and apolipoproteins.

Glycosylated lipoproteins have been suggested to be the subject of further processing leading to the formation of hyperglycosylated compounds. Glycosylation and hyperglycosylation of proteins including lipoproteins in both plasma and the arterial wall can also be expected to be a risk factor for cardiovascular disease including arteriosclerosis, also in diabetic subjects. Accordingly, a composition according to the present invention may be capable of preventing, treating, alleviating, reducing and/or eliminating the accumulation of hyperglycosylated proteins in both serum and cells of the arterial wall. By doing so, the composition is acting to decrease the amount of LDL becoming ’’trapped” in the arterial wall due to the high degree of glycosylation of arterial wall proteins. A composition according to the present invention may also be effective in preventing and/or alleviating any change to the endothelial cell wall that increase LDL ’’trapping”, and it may be effective in restoring the formation of cells with normal permeability and adhesion parameters.

Lipoprotein glycosylation, hyperglycosylation, oxidation and/or auto-oxidative glycosylation, are risk factors for cardiovascular disease such as arteriosclerosis, including arteriosclerosis in diabetes. Accordingly, a composition according to the present invention may be effective in preventing, treating, alleviating, eliminating and/or reducing any incidence of lipoprotein glycosylation, hyperglycosylation, oxidation and/or auto-oxidative glycosylation. According to one presently preferred hypothesis, the phytoestrogen compound of a composition according to the present invention is capable of counteracting such incidences. The phytoestrogen compound may also be capable of preventing, reducing and/or eliminating the formation of e.g. free radicals that are likely to be involved in such processes, and a composition according to the present invention may be effective in being, promoting, and/or facilitating the formation of an effective antioxidant defense system for counteracting glycosylation, hyperglycosylation, oxidation and/or auto-oxidative glycosylation of serum proteins and proteins including lipoproteins of the arterial cell wall.

Since oxidative stress is a characteristic of diabetes and possibly a contributory factor to among others lipoprotein oxidation and/or glycosylation, and since no efficient antioxidant protection exists due to e.g. significantly decreased levels in diabetic subjects of antioxidants such as e.g. ascorbic acid, a composition according to the present invention may be effectively acting as an antioxidant in preventing lipoprotein oxidation and/or glycosylation.

Generally, a composition according to the present invention may be effectively acting as an antioxidant in preventing lipoprotein oxidation and/or glycosylation. By the term auto-oxidative glycosylation, or glycoxidation, is understood a reaction catalyzed e.g. by reducing sugars that leads to an oxidative modification and/or cross-linking of proteins. The rate of such a process can be expected to be increased in the presence of high glucose concentrations since the oxidizing potential is significantly increased under such circumstances. An increased production of free radicals and lipid peroxidation may also contribute to the formation of auto-oxidative glycosylated lipoproteins and this contribution may also be effectively prevented and/or eliminated by a composition according to the present invention.

According to another presently preferred hypothesis, the binding of phytoestrogen compounds such as e.g. isoflavones, optionally in combination with soy peptides e.g. obtainable by partial hydrolysis of soy protein, to a receptor in the arterial wall, such as e.g. the estrogen receptor, or an estrogen-like receptor, and optionally influenced by the level of specific phospholipids, such as phosphatidyl choline, is involved in or effective in controlling accumulation of lipoproteins and uptake of cholesterol and/or triglycerides in the arterial wall, possibly by regulating the permeability of said wall and/or the mechanism of lipoprotein and/or cholesterol and/or triglyceride transport across cellular membranes. Consequently, the binding of isoflavones such as e.g. genistein and/or daidzein to a receptor in the arterial wall may reduce the flux of lipoproteins into the arterial wall and/or prevent cholesterol and/or triglycerides from entering the arterial wall, or reduce and/or substantially eliminate the amount of cholesterol and/or triglycerides that enters the arterial wall. Receptor binding of isoflavones in the arterial wall is particularly effective in controlling, preventing and/or eliminating fatty streak formation and/or fibrous plaque development and/or effective in mediating a regression of one or both of said arteriosclerotic conditions.

Both isoflavones and the amino acid arginine which is abundant in soy protein, release nitric oxide (NO), which relaxes muscle cells in arteries and other smaller blood vessels, leading to vasodilation and thereby increased blood and oxygen flow.

Isoflavones also provide strong anti-inflammatory effects. According to a particularly preferred hypothesis, binding of an isoflavones such as e.g. genistein and/or daidzein to a receptor in the arterial wall, preferably an estrogen receptor or an estrogen-like receptor, results in an increased nitric oxide synthesis in the endothelial cells of the arterial wall. Nitric oxide is known to exert anti- arteriosclerotic effects including inhibition of platelet adhesion and aggregation, and inhibition of smooth muscle cell proliferation. Soy peptides and the amino acid arginine obtainable by hydrolysis of soy protein may participate in the binding of isoflavones to an estrogen receptor or an estrogen-like receptor or the soy peptides may themselves bind to said receptor and exert an action leading to an increased nitric oxide synthesis. Furthermore, specific phospholipids, e.g. phosphatidyl choline, may in response to cell stimuli, be broken down by specific phospholipases, thereby resulting in a number of hydrolysis products having the potential to act as second messengers and thus markedly influence the effect of phytoestrogens.

In another presently preferred hypothesis, the establishment of an oxidative potential that promotes lipoprotein oxidation and/or lipoprotein auto-oxidative glycosylation occurs concomitantly with, and is very likely caused by, a decrease in cellular antioxidative defense systems. This hypothesis is supported by the fact that e.g. ascorbic acid concentrations are decreased in many diabetic individuals. Accordingly, a composition according to the present invention may be effective in acting as an antioxidant. This action reduces and/or eliminates LDL, VLDL, IDL and/or HDL susceptibility to oxidation. Concomitantly with a direct anti-oxidative effect, a composition according to the present invention may also lower the increased serum glucose levels and by doing so, a composition according to the present invention may be effective in reducing the oxidizing potential causing and/or contributing to oxidative stress.

Furthermore, a composition according to the present invention may also be effective in reducing an enhanced susceptibility to endothelial injury and/or for alleviating and/or restoring and/or improving an inefficient endothelial cell repair mechanism leading to endothelial dysfunction. One effect of such an action exerted by a composition according to the present invention is to direct macrophage development away from foam cell formation and to increase the potential of generating arterial smooth muscle cells. The development of incapacitating neurological diseases like dementia and Alzheimer’s disease have generally been associated with a wide range of histological and pathophysiological changes, of which the possible contribution of cerebrovascular deficiencies has also been suggested. Thus, it has been hypothesised that various forms of cerebrovascular insufficiency such as reduced blood supply to the brain or disrupted microvascular integrity in cortical regions may occupy an initiating or intermediate position in the chain of events ending with a setback leading to neuronal damage and a concomitant suboptimal cognitive capacity.

The small vessels of the cerebrovascular system can broadly speaking be said to comprise two components. First, the leptomeninges vasoganglion, and second, the perforating arteries derived from anterior, middle, posterior cerebral arteries that supply the subcortical parenchyma. All cerebral small vessels are crucial to maintenance of adequate blood flow to the sub-surface brain structure and include small arteries, arterioles, venules, and capillaries.

Clinically what is known as cerebral small vessel disease (CSVD), and which affects the small arteries, arterioles, venules, and capillaries of the brain, refers to several cerebrovascular deficiencies. Thus, small vessel disease is believed to account for up to 25% of all ischemic strokes and is a leading cause of functional loss, disability and cognitive decline in the elderly. The main clinical manifestations of CSVD include stroke, cognitive decline, dementia, psychiatric disorders, abnormal gait, and urinary incontinence.

Among the pathologic changes involved in CSVD, the two most common are arteriolosclerosis and cerebral small vascular atherosclerosis. Cerebral small vascular atherosclerosis is inter alia characterized by proliferation of fibroblasts, deposits of fibro-hyaline material and collagens, thickening of the vessel wall, formation of microatheroma, and a narrowing of the lumen of the small arteries, arterioles, venules, and capillaries of the brain. In addition, vessel wall damage causes distension of its outer portions, due to fibrosis, and stenosis or obstruction of the proximal lumen.

Critical stenosis and hypoperfusion involving multiple small arterioles, mainly in deep white matter, lead to incomplete ischemia which are visualized as White Matter Hyperintensities on neuroimaging. Conventional risk factors such as hypertension diabetes, smoking, high homocysteine concentrations, obesity, and dyslipidemia have been considered to lead to arteriolosclerosis White Matter Hyperintensities, the presence of which has, furthermore, been correlated with the presence of unstable carotid plaques.

The clinical manifestations of CSVD vary depending on the specific cause of the disease, as well as the brain regions affected. Thus, individuals may either present sudden onset stroke symptoms or symptoms of progressive cognitive deterioration, dementia, Alzheimers disease or the like. In fact CSVD is thought to be among the main causes of vascular cognitive impairment, and is thought to account for about 45% of dementia cases.

In accordance with the fact that arteriolosclerosis, is known to be age-related, and is the most common small vessel alteration in aged brains, the severity of arteriolosclerosis increases with aging. Risk factors for CSVD include hypertension, cigarette smoking, diabetes, atrial fibrillation, hyperhomocysteinemia, chronic kidney disease, high circulatory phosphate level and obesity. Thus, controlling or removing these known risk factors is important for the prevention of CSVD. Hypertension is the most prevalent and important risk factor and many studies have shown that lowering blood pressure reduces stroke and dementia or cognitive decline in patients who have had a stroke and in those without cerebrovascular disease. Also, improving vasomotor reserve capacity and cerebral endothelial function by inhibiting cerebral vascular superoxide production, enhancing endothelial-derived Nitric Oxide bioavailability and decreasing oxidative stress, could protect against cerebral ischemia.

In addition, evidence is fast accumulating which indicates that Alzheimer's disease is indeed primarily a vascular disorder with neurodegenerative consequences rather than a neurodegenerative disorder with vascular consequences.

Thus, in essence primarily two factors need to be present for Dementia or Alzheimer’s disease to develop: (1) advanced ageing, and (2) presence of a condition that lowers cerebral perfusion, i.e. a vascular-risk factor. The first factor (advanced ageing) introduces a normal but potentially insidious process that lowers cerebral blood flow in inverse relation to increased ageing; the second factor (a vascular risk factor) adds a crucial burden which further lowers brain perfusion. Convergence of these two factors will culminate in a critically attained threshold of cerebral hypoperfusion, which will destabilize neurons, synapses, neurotransmission and cognitive function, creating in its wake a neurodegenerative state characterized by the formation of White Matter Hyperintensities/plaques, neurofibrillary tangles, amyloid angiopathy and in some cases, Lewy bodies. At the same time any development of Alzheimer’s disease or dementia will be augmented by distortion of the regional brain capillary structure and the impairment of nitric oxide (NO) release which will again influence the vascular and neural systems. Thus, the clinical convergence of advanced ageing in the presence of a chronic, pre-morbid vascular risk factor, will, with time, contribute to an endotheliopathy involving basal NO deficit leading to a progressive neurodegeneration, which is characteristic of dementia and Alzheimer's disease. Also, recent research, in the form of a retrospective cohort study including participants of European ancestry aged at least 60 years without dementia at baseline, has shown that a healthy lifestyle is associated with lower risk of dementia regardless of the genetic risk. Thus, among older adults without cognitive impairment or dementia, both an unfavorable lifestyle and a high genetic risk was in this study shown to be significantly associated with higher dementia risk, whereas a favorable lifestyle was shown to be associated with a lower dementia risk even in individuals with high genetic risk (Association of Lifestyle and Genetic Risk With Incidence of Dementia, llianna Lourida; Eilis Hannon; Thomas J. Littlejohns; et al, JAMA. 2019;322(5):430- 437).

Consequently, a composition according to the present invention may be used in preventing, treating and/or alleviating Alzheimer disease and/or Dementia. The unique dyslipidemia associated with type 2 diabetes is a major risk factor for cardiovascular disease, and prevention, alleviation, reduction and/or elimination of dyslipidemia in diabetic subjects is a prime objective of administration of a composition according to the present invention to a diabetic individual. Another important objective of such an administration is the development in a diabetic subject of a gradually reduced insulin resistance and/or a gradually improved glucose tolerance. Since increasing insulin resistance and impaired glucose tolerance are key elements in the progression of type 2 diabetes, the same factors most also be a natural focus of any preventive treatment.

In another presently preferred hypothesis, a composition according to the present invention will promote and/or mediate a reduction in arterial wall thickness and lead to a reduction in the amount of LDL entering the wall. It is believed that an increased thickness of the arterial wall is positively associated with an increased uptake of LDL particles that are likely to either aggregate or oxidize within the cells of the arterial wall. Also, a composition according to the present invention may be capable of reducing, eliminating and/or preventing the formation of increased serum levels of lipoprotein (a) in a subject, including a diabetic subject. Lipoprotein (a) levels may primarily be genetically determined, and no current cardiovascular medications are thought effective in lowering serum levels of lipoprotein (a).

Obstructive pulmonary disease (OPD) including chronic obstructive pulmonary disease (COPD) as used herein is defined as a condition comprising subjects with airways limitations or obstructions or subjects with a mucus hypersecretory condition including chronic mucus hypersecretion, i.e. subjects with asthma including chronic asthma and subjects with bronchitis including chronic bronchitis. However, a clear distinction between e.g. bronchial asthma and chronic bronchitis can be difficult and sometimes impossible to make, and a sharp distinction between COPD and OPD is therefore not always possible.

Mucus hypersecretion and a limited or obstructed airflow are two major characteristics of COPD. According to one presently preferred theory, mucus hypersecretion is an initial mechanism that leads to recurrent respiratory infections, that in turn generates a destruction of the airways and promotes a development of pulmonary parenchyma and airflow obstruction. At least two separate conditions, i) mucus hypersecretion and ii) dyspnea, are identifiable due to an obstructive or limited lung function. Chronic mucus hypersecretion and obstructive airflow are not necessarily related, since an individual may have a hypersecretory disorder only, or an obstructive disorder only, or both a hypersecretory and an obstructive disorder. Chronic mucus hypersecretion is associated with an impaired mucociliary clearance and may therefore predispose to lung cancer by causing a prolonged contact between potential carcinogens with the bronchial epithelium. Accordingly, a composition according to the present invention may be effective in treating and/or alleviating mucus hypersecretion and dyspnea in a subject.

Asthma as used herein is defined as a respiratory disease in which spasm and constriction of the bronchial passages and swelling of their mucous lining cause obstruction of breathing, often, but not exclusively, due to allergy. One mechanism for expiratory airflow limitation in asthma is a smooth muscle contraction leading to a narrowing of the airway lumen. Asthma is frequently divided clinically into extrinsic and intrinsic asthma, separating asthma triggered by environmental allergens from that in which atopy does not appear to play a major role. Consequently, a composition according to the present invention may be effective in preventing, treating and/or alleviating smooth muscle contraction.

In asthma the airways are occluded by tenacious plugs of exudate and mucus, and there occurs a fragility of airway surface epithelium, thickening of the reticular layer beneath the epithelial basal lamina, bronchial vessel congestion and edema. An increased inflammatory infiltrate comprising "activated" lymphocytes and eosinophils, and an enlargement of bronchial smooth muscle, particularly in medium-sized bronchi, is also observed. Asthma comprises at least extrinsic (atopic or allergic) and intrinsic (non-atopic) divisions, each of which present clinically in a variety of ways. A composition according to the present invention may be effective in preventing and/or alleviating the formation of tenacious plugs of exudate and mucus, effective in preventing, treating and/or alleviating a fragility of airway surface epithelium subsequently generated by mucus secretion, effective in preventing, reducing and/or eliminating any thickening of the reticular layer beneath the epithelial basal lamina, and effective in preventing, treating and/or alleviating bronchial vessel congestion and/or edema.

Asthma may in some cases be regarded as a chronic inflammatory disease. Since the term chronic asthmatic bronchitis has no clearly defined pathologic equivalent, patients having a chronic productive cough normally associated with chronic bronchitis, as well as bronchospasms, at the same time as having an airflow obstruction, will be regarded as suffering from both chronic bronchitis as well as small airways disease (chronic obstructive bronchitis) and asthma, since the pathology presumably would be that of those conditions.

A composition according to the present invention may be effective in preventing, alleviating and/or curing inflammation of the airways, whether transient or chronic.

Airway inflammation is thought to be an important contributor to asthma, and airway inflammation may well be present even in the absence of severe symptoms of asthma.

In one particularly preferred aspect the present invention provides a treatment and/or alleviation of an inflammation of the airways by means of an anti-oxidative effect exerted by a composition according to the present invention. The anti-oxidative effect may in particular be exerted by the naturally occurring isoflavones forming part of a composition according to the present invention. A composition according to the present invention may be effective in increasing FEVi, as measured by forced expiratory volume in the first second of expiration, said effect being exerted by the binding of a component of the composition, particularly naturally occurring isoflavones, to beta-2 receptors or receptors belonging to the class of beta-2 receptors. Beta-2 receptors are present on many different types of cells including cells in airways and vessels. A composition according to the present invention may also be effective in generating a dilatation of the airways in a subject, preferably a subject suffering from a pulmonary disease.

The occurrence of bronchial inflammation in asthma is, according to one presently preferred hypothesis, thought to arise at least in part from an airway response to an antigen in an allergic subject. The response includes immediate pulmonary mast-cell activation and initiation of an inflammatory response that develops over hours and is important in the later and more persistent development of bronchial obstruction. A composition according to the present invention may be effective in treating, alleviating and/or eliminating several of the causes of airway obstruction that - alone or in combination - contributes to bronchial hyperresponsiveness, i.e. the fundamental defect in asthma. Importantly, airway inflammation is believed to be a crucial component for i) the chronicity of asthma, ii) the intensity of airways hyperresponsiveness, and iii) the absence of a complete therapeutic control, when bronchodilator therapy is used alone. Consequently, a composition according to the present invention may be effective in controlling, reducing and/or eliminating edema, mucus secretion, and inflammation of the airways resulting at least in part from a response to an allergen.

Bronchitis as used herein is defined as an acute or chronic inflammation of any part of the bronchi and bronchial tubes. The bronchi are large delicate tubes in the lungs that are attached to the trachea and carry air to smaller tubes in the lungs. In bronchitis, including chronic bronchitis, there is mucous hypersecretion, an enlargement of tracheobronchial submucosal glands, and a disproportionate increase of mucous acini. Acute bronchitis is often characterized by fever, chest pain, severe coughing, and secretion of mucous material coughed up from the respiratory tract. Acute bronchitis affects the branches of the bronchi and may develop into bronchial or lobular pneumonia. Chronic bronchitis may result from repeated attacks of acute bronchitis. Consequently, a composition according to the present invention may be effective in controlling mucous hypersecretion, preventing, treating and/or alleviating an enlargement of tracheobronchial submucosal glands, and reduce and/or eliminate a disproportionate increase of mucous acini. The pathologic equivalent to chronic bronchitis is a non-specific series of changes in the bronchial wall generally characterized by an increase in the size and number of mucous glands and an increased number of goblet cells in the epithelium. When progressing into a chronic condition, bronchitis is a serious and incurable disorder. Consequently, a composition according to the present invention may be effective in controlling a series of changes in the bronchial wall generally characterized by an increase in the size and number of mucous glands and an increase in the number of goblet cells. A composition according to the present invention may also be capable of reducing and/or eliminating any mucos production including an increased mucos production.

Bronchial infections usually remain confined to the mucosa, and some resolve spontaneously without the need for treatment. Chronic bronchitis affects both the large and small airways. In the large airways, hypertrophy and hyperplasia of glandular structures and goblet cell metaplasia are prominent features of the condition. In the small airways, peribronchiolar fibrosis and airway narrowing may be prominent features. In chronic bronchitis hypertrophy of glandular structures and goblet cell metaplasia in the proximal airways likely contribute to an increased mucus production, the expectoration of which is one defining characteristic of chronic bronchitis. Consequently, a composition according to the present invention may be effective in preventing an airflow limitation in a subject prone to contracting bronchitis and/or to alleviate any airflow limitation or obstruction already present in said subject.

Particularly, a composition according to the present invention may be effective in controlling hypertrophy and hyperplasia of glandular structures and goblet cell metaplasia, as well as peribronchiolar fibrosis and a narrowing of the small airways.

Bronchitis may be caused by a number of factors including viral and/or bacterial infection, environmental pollutants including cigarette smoke, and allergy. These factors may occur together or separately. A viral infection may e.g. predispose an individual to a subsequent bacterial infection. Bronchial infections occur in patients with abnormal airways who have reduced host defenses. The three major bacterial pathogens isolated during bronchial infections are non-typable Haemophlus influenzae, moraxella catarrhalis, and Streptococcus pneumoniae. A composition according to the present invention may especially be effective in preventing viral and/or bacterial infection in a subject by e.g. increasing the host defenses of said subject.

The term small airways as used herein is defined as small bronchi and bronchioles that contain no cartilage, glands or alveoli in their walls and measure 2 mm or less in internal diameter. The term small airways disease is used for a group of non-specific histological changes of peripheral airways found in individuals with a limited or obstructed airflow, including individuals having features such as mucus plugging, chronic inflammation, and muscular enlargement of small airway walls. Small airways disease is present in some patients with the clinical picture of chronic bronchitis. Consequently, a composition according to the present invention may be effective in preventing, treating, prophylactically treating and/or alleviating a limited or obstructed flow of air through the small airways.

In small or peripheral airways disease, there is inflammation of bronchioli and mucous metaplasia and hyperplasia, increased intraluminal mucus and increased wall muscle. Consequently, a composition according to the present invention may be effective in controlling inflammation of the bronchioli and mucous metaplasia and hyperplasia, and effective in reducing and/or eliminating any increased intraluminal mucus formation and/or any increased wall muscle development.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows changes in LDL cholesterol (LDL-C) from baseline after 8 weeks, as reported in Example 6.

Figure 2 shows changes in total cholesterol (Total-C) from baseline after 8 weeks, as reported in Example 6.

Figure 3 shows changes in apolipoprotein B from baseline after 8 weeks, as reported in Example 6.

Figure 4 shows changes in triglycerides from baseline after 8 weeks, as reported in Example 6.

Figure 5 shows lipid lowering effects of the composition according to the present invention and isolated soy protein (ISP), as reported in Example 6.

Figure 6 shows superior lipid lowering effects of the composition according to the present invention compared to isolated soy protein (ISP), as reported in Example 6.

Figure 7 shows the changes in aortic wall cholesterol nmol/mg from baseline after 75 days as reported in Example 5. DETAILED DESCRIPTION OF THE INVENTION

A composition according to the present invention comprises a novel combination of soy protein, preferably isolated soy protein, a phytoestrogen compound in the form of naturally occurring isoflavones, and optionally a phospholipid source, preferably having high fixed levels of phosphatidyl choline, more preferably soy lecithin and optionally dietary fibers, preferably soybean fibers, more preferably soy cotyledon fibers.

The soy protein can be provided by isolated soy protein, soy protein concentrate, soy flour or the like or any combination thereof. Isolated soy protein is preferred.

Processed Isolated soy protein is particularly preferred.

Isolated soy protein is the major proteinacious fraction of soybeans. It is prepared from high quality, dehulled, defatted soybeans by removing a preponderance of the non protein components resulting in an isolated soy protein fraction which in the present context shall contain at least 90 percent protein (N x 6.25) on a moisture free basis. The preparation takes place through a series of steps in which the soybean protein portion is separated from the rest of the soybean. The removal of carbohydrate results in a product, which is essentially bland in flavor and therefore particularly useful in a nutritional composition for humans.

Soy protein concentrates are made by removing most of the oil and water-soluble non protein constituents from defatted and dehulled soybeans. In the present context a soy protein concentrate shall preferably contain at least 65 percent protein on a moisture- free basis.

The soy protein can also be provided by soy flour, which can be full-fat or defatted soy flour. Full-fat soy flour comes from whole, dehulled soybeans that have been ground into a fine powder and, as the name implies, still contains the fat naturally found in soybeans. Defatted soy flour comes from whole, dehulled, defatted soybeans that have been ground into a fine powder. Soy flour is approximately 50 percent soy protein on a dry weight basis in the present context.

The soy protein used in a composition according to the present invention should preferably supply all the essential amino acids in the amounts required for humans. Preferably, the soy protein should also meet or exceed the essential amino acid requirement pattern for children and adults as established by the Food and Agricultural Organization, World Health Organization and United Nations University (FAO/WHO, UNU). Furthermore, the preferred soy protein should be comparable in digestibility to milk, meat, fish, and egg protein. Finally, the preferred soy protein shall be effective in maintaining nitrogen balance when consumed at the recommended protein intake level.

Researchers have shown that specific amino acids may to some extent effect serum lipid levels and potentially alleviate cardiovascular diseases. Animal studies have indicated that the amino acid lysine increases serum cholesterol levels, while arginine counteracts this effect (Kurowska et al., J. Nutr. 124, 364-370 (1994) and Sanchez et al., Med. Hypotheses 35, 324-329 (1991). This observation appears to be in correspondence with the well established influence of NO on vasodilation, since arginine may potentially be converted to citrullin and NO by NO-synthetase. Thus, the high level of arginine in soy protein, as well as natural soy isoflavones, may contribute to effectively increasing the levels of NO (nitric oxide), which would significantly increase blood flow through vasodilatation. This effect on increased blood flow is also found in capillaries (see e.g., “L-Arginine Induces Nitric Oxide-Dependent Vasodilation in Patients With Critical Limb Ischemia, A Randomized, Controlled Study”, originally published Jan 1996 https://doi.Org/10.1161/01.CIR.93.1.85, Circulation 1996;93:85- 90). Thus, according to a presently preferred hyphothesis soy protein having a high arginine to lysine ratio effects serum lipid levels and alleviates symptoms of cardiovascular diseases to a greater extent than soy protein having a lower or normal arginine to lysine ratio, just like it induces NO-dependent peripheral vasodilation (see e.g. “A Meal Enriched with Soy Isoflavones Increases Nitric Oxide-Mediated Vasodilation in Healthy Postmenopausal Women”, Wendy L. Hall, Nichola L. Formanuik, Duangporn Harnpanich, Monique Cheung, Duncan Talbot, Philip J. Chowienczyk, Thomas A. B. Sanders, The Journal of Nutrition, Volume 138, Issue 7, July 2008, Pages 1288-1292, https://doi.Org/10.1093/jn/138.7.1288). Consequently, soy protein having a high arginine to lysine ratio is a particularly preferred soy protein source in a composition according to the present invention. Preferably the soy protein of the soy protein source in a composition according to the present invention should have an arginine to lysine ratio of at least about 1.0, such as at least about 1.1, for example at least about 1.2, such as at least about 1.3, for example at least about 1.4, such as at least about 1.5, for example at least about 1.6, such as at least about 1.7, for example at least about 1.8, such as at least about 1.9, for example more than about 2, such as at least about 2.1 , for example at least about 2.2, such as at least about 2.5, for example at least about 2.75, such as at least about 3, for example more than about 3.3, such as at least about 3.6, for example at least about 4, such as at least about 4.5, for example at least about 5, such as at least about 6, for example at least about 7, such as at least about 8, for example at least about 9, such as at least about 10, for example at least about 11, such as at least about 12, for example at least about 13, such as at least about 14.

The soy protein is preferably the main or sole protein source in a nutritional composition according to the present invention. However, parts of the protein source may be provided by other proteins such as e.g. skimmed milk, preferably as a powder, and other vegetable or animal proteins including dairy proteins. Preferably, at least 50 weight percent, for example at least 60 weight percent, such as at least 70 weight percent, for example at least 75 weight percent, such as at least 80 weight percent, for example at least 85 weight percent, such as at least 90 weight percent, for example at least 95 weight percent, such as at least 98 weight percent of the total protein content of the composition is soy protein, and more preferably substantially all of the protein is soy protein.

In a preferred embodiment of the invention the soy protein is provided by isolated soy protein. In this embodiment, preferably at least 50 weight percent, for example at least 60 weight percent, such as at least 70 weight percent, for example at least 75 weight percent, such as at least 80 weight percent, for example at least 85 weight percent, such as at least 90 weight percent, for example at least 95 weight percent, such as at least 98 weight percent of the total protein content of the composition is isolated soy protein, and more preferably substantially all of the protein is provided by isolated soy protein.

The total protein content of a composition according to the present invention provides at least 15 percent of the total energy content of the composition, for example 18 percent, such as at least 20 percent, for example at least 22 percent, such as at least 25 percent, for example at least 28 percent, such as at least 30 percent, for example at least 32 percent, such as at least 35 percent, for example at least 38 percent, such as at least 40 percent, for example at least 42 percent, such as at least 45 percent, for example at least 48 percent, such as at least 50 percent of the total energy content of the composition, and preferably less than 90 percent of the total energy content of the composition.

Phytoestrogen compounds according to the present invention are naturally occurring plant substances, said substances being either structurally or functionally similar to 17p-estradiol or generating estrogenic effects. Phytoestrogens consist of a number of classes including isoflavones, coumestans, lignans and resorcylic acid lactones. Examples of isoflavones according to the present invention are genistein, daidzein, equol, glycitein, biochanin A, formononetin, and O-desmethylangolesin. The phytoestrogen compounds of a composition according to the present invention are naturally occurring isoflavones, more preferably genistein, daidzein and/or glycitein, yet more preferably genistein and/or daidzein, and even more preferably genistein. A preferred composition according to the present invention may accordingly comprise a single isoflavone, such as genistein, daidzein or glycitein, or it may comprise at least one isoflavone selected from the group comprising at least genistein, daidzein and glycitein. When present in the plant the isoflavones are mainly in a glucoside form, i.e. attached to a sugar molecule. This glucoside form can be deconjugated to yield a so- called aglycone form, which is the biologically active compound. A composition according to the present invention may comprise isoflavones in glucoside and/or aglycone forms regardless of whether the deconjugation to the aglycone form has taken place biologically, in vitro or by any other means whereby the isoflavones are included in a composition according to the present invention or if the aglycone forms are the native form of the isoflavones.

A number of studies have shown that a decrease in bone mineral content (BMC) may be a result of weight loss, menopause or exercise. The reduction in BMC may be due to a reduced mineral intake, but can also partially be attributed to the hormonal changes associated with e.g. strict dieting and weight loss or menopause, as the decrease of estrogen in both males and females is associated with osteoporotic changes. The decrease in BMC may be counteracted, to a certain degree, by the intake of calcium supplements, but in the literature several lines of evidence suggest that also soy isoflavones and related compounds act as estrogen agonists and have beneficial related skeletal effects. A beneficial effect of ingestion isoflavones have been demonstrated by a 2-year study, performed in Denmark at Institute for Optimum Nutrition, in which it could be shown that isoflavone-rich soymilk (100 mg/d) prevents bone loss in the lumbar spine of postmenopausal woman (Eva Lydeking-Olsen et al.). This study is of specific interest because there are no previously published studies on the long term effect of soy intake on bone mass, only short term human studies have shown a bone-sparing effect of soy protein with isoflavones in the range of 80-90 mg/d. Thus, without being bound by theory, a composition according to the present invention preferably contains an amount of isoflavones capable of providing a beneficial effect on the BMC.

The naturally occurring phytoestrogen compound in the form of soy isoflavones is preferably present in an amount of at least about 0.34 weight percent of the soy protein content. More preferably the phytoestrogen compound is present in an amount of at least 0.34 weight percent, such as at least about 0.35 weight percent, for example at least about 0.40 weight percent, such as at least about 0.45 weight percent, for example at least about 0.50 weight percent, such as at least about 0.55 weight percent, for example at least about 0.60 weight percent, such as at least about 0.65 weight percent, for example at least about 0.70 weight percent, such as at least about 0.75 weight percent, for example at least about 0.80 weight percent, such as at least about 0.85 weight percent, for example at least about 0.90 weight percent, such as at least about 1.0 weight percent of the soy protein content, and preferably less than 2.50 weight percent of the soy protein content.

In the past, the downstream processing techniques used in the preparation of soy proteins have included steps that removed and/or destroyed isoflavones. Methods are available today, which provide soy protein products with high, fixed levels of naturally occurring isoflavones. The isoflavones according to the present invention in glucoside and/or aglycone forms can be included in a composition according to the present invention as part of such soy protein products and/or by themselves and/or as part of any other composition comprising isoflavones.

A substantial part of the naturally occurring isoflavones of a composition according to the present invention are provided in the form of a soy protein source with a high concentration of naturally occurring isoflavones and/or soy germ, and not in the form of isoflavones in an isolated extracted form. Without wishing to be bound by theory it is currently a preferred hypothesis that the published conflicting evidence regarding the clinical effects of isoflavones in extracted form may be explained by the fact that isoflavones are negatively affected or compromised by the harsh extracting techniques used when isolating these compounds from the original plant material using e.g. chemical-, ethanol or UHT extraction, such as e.g. the methods described in e.g. US 5 858449. Thus, it is contemplated that a detrimental effect of processing soy protein, including also a loss of the otherwise well-documented cholesterol lowering effect (even reversed to a negative effect on total and LDL cholesterol), is an inherent result of using e.g. UHT processed or otherwise processed soy protein to arrive at isoflavone-enriched soy protein products, such as the products described in e.g. US 5 858449. This hypothesis would also appear to be in line with the fact that soy germ rich in isoflavones seems to give rise to a significant reduction of cardiovascular risk factors and improved endothelial function in patients with Type 2 Diabetes (see e.g., https://www.ncbi.nlm.nih.gov/pubmed/21788625/ Diabetes Care. 2011 Sep;34(9): 1946-8. doi: 10.2337/dc11-0495. Epub 2011 Jul 25. “Novel soy germ pasta improves endothelial function, blood pressure, and oxidative stress in patients with type 2 diabetes”). Thus, a soy germ based pasta gave rise to a significantly improved arterial stiffness (P = 0.005) and reduced systolic (P = 0.026) and diastolic (P = 0.017) blood pressures. Plasma TAC increased (P = 0.0002), oxidized LDL cholesterol decreased (P = 0.009), 8-iso-PGF2a decreased (P = 0.001), GSH levels increased (P = 0.0003), and homocysteine decreased (P = 0.009) consistent with a reduction in oxidative stress, whereas no significant changes were observed with conventional pasta.

The optional phospholipid source according to the present invention will preferably comprise polyunsaturated fatty acids and monounsaturated fatty acids and optionally also saturated fatty acids. Soy lecithins and a-linolenic acid are particularly preferred.

The optional phospholipid source will preferably comprise at least about 5% phosphatidyl choline, such as at least 10% phosphatidyl choline. The optional phospholipid source will more preferably comprise at least about 20% phosphatidyl choline, such as at least about 30% phosphatidyl choline, for example at least about 35% phosphatidyl choline, such as at least about 40% phosphatidyl choline, for example at least about 45% phosphatidyl choline, such as at least about 50% phosphatidyl choline, for example more than about 55% phosphatidyl choline phosphatidyl choline by weight, such as at least 60% phosphatidyl choline, for example at least about 65% phosphatidyl choline, such as at least about 70% phosphatidyl choline, for example at least about 71% phosphatidyl choline, such as at least about 72% phosphatidyl choline, for example at least about 73% phosphatidyl choline, such as at least about 74% phosphatidyl choline, for example more than about 75% phosphatidyl choline, such as at least about 76% phosphatidyl choline, for example at least about 77% phosphatidyl choline, such as at least about 78% phosphatidyl choline, for example at least about 79% phosphatidyl choline, for example more than about 80% phosphatidyl choline, such as at least about 85% phosphatidyl choline, for example at least about 90% phosphatidyl choline, such as at least about 98% phosphatidyl choline, for example 100% phosphatidyl choline by weight.

The optional phospholipid source will preferably comprise polyunsaturated fatty acids and monounsaturated fatty acids and optionally also saturated fatty acids. The amount of polyunsaturated fatty acids and monounsaturated fatty acids, including the essential fatty acids, may range from 35 to 50, preferably 38 to 44, weight percent of the total amount of the fat source. The essential fatty acids are also called omega-6 and omega-3 fatty acids and include linolic acid and/or linolenic acid (a-linolenic acid). The amount of saturated fatty acids may be from 20 to 30 weight percent, preferably 22 to 26 weight percent, of the total amount of the phospholipid source. In a composition according to the present invention, the optional phospholipid source usually provides from 5 to 70 percent, preferably 10 to 60 percent, such as from 15 to 50 percent, for example from 20 to 40 percent, such as from 25 to 35 percent of the total energy content of the composition.

The optional phospholipid source preferably provides at least about 5 percent of the total energy content of the composition, such as at least about 10 percent, for example at least about 15 percent, such as at least about 20 percent, for example at least about 21 percent, such as at least about 22 percent, for example at least about 23 percent, such as at least about 24 percent, for example more than about 25 percent, such as at least about 26 percent, for example at least about 27 percent, such as at least about 28 percent, for example at least about 29 percent, such as at least about 30 percent, for example more than about 31 percent, such as at least about 32 percent, for example at least about 33 percent, such as at least about 34 percent, for example at least about 35 percent, such as at least about 36 percent, for example at least about 37 percent, such as at least about 38 percent, for example at least about 39 percent, such as at least about 40 percent, for example at least about 45 percent, such as at least about 50 percent, for example at least about 55 percent, such as at least about 60 percent, for example at least about 65 percent of the total energy content of the composition, and preferably less than 70 percent of the total energy content of the composition. The optional dietary fibers used in a presently preferred embodiment of the present invention should preferably comprise a mixture of insoluble fibers and water-soluble fibers also referred to as soluble fibers. Soluble fibers have a lowering effect on blood cholesterol levels. Examples of dietary fibers comprising soluble fibers are fibers from apples, bananas, oranges, carrots, oats, and soybeans. The dietary fibers preferably comprise soluble fibers in an amount of about 5 weight percent, such as about 10 weight percent, for example about 15 weight percent, such as about 20 weight percent, for example about 25 weight percent, such as about 30 weight percent, for example about 35 weight percent, such as about 40 weight percent, for example about 45 weight percent, such as about 50 weight percent, for example about 55 weight percent, such as about 60 weight percent, for example about 65 weight percent, such as about 70 weight percent, for example about 75 weight percent, such as about 80 weight percent, for example about 85 weight percent, such as about 90 weight percent, for example about 95 weight percent. The dietary fibers optionally used in the present invention are preferably soybean fibers, more preferably soy cotyledon fibers. Such fibers are derived from dehulled and defatted soybean cotyledon and are comprised of a mixture of soluble and insoluble fibers. Soy cotyledon fibers are distinctly different from soybean fibers derived from soy hulls as well as other fiber sources. Soy cotyledon fibers are bland tasting, contain no cholesterol, are low in fat and sodium, and they have good water-binding properties and low caloric content.

Soy cotyledon fibers supplied in a fat-modified and low-cholesterol diet are known to further reduce serum cholesterol levels in a subject suffering from mild to severe hypercholesterolemia. The effect is a lowering of the serum levels of total cholesterol including a lowering of the serum levels of LDL-cholesterol. However, HDL-cholesterol and total triglycerides are not significantly affected by soy cotyledon fibers. Soybean fibers, in particular soy cotyledon fibers, are believed to provide a synergistic effect in combination with soy protein and/or with a phytoestrogen compound, such as naturally occurring isoflavones, or to exert a potentiating effect on the soy protein and/or the phytoestrogen compound, said synergistic or potentiating effect being effective in lowering serum levels of lipid and cholesterol in subjects having normal as well as elevated serum levels of total cholesterol and total triglycerides.

Without wishing to be bound by any specific theory it is presently believed that both soluble dietary fibers (working as nutrients) and insoluble dietary fibers (working as bulking agents), in particular from soybean fibers, more particularly soy cotyledon fibers, provide favorable growth conditions for the microflora in the human gut, which makes the microflora more effective in deconjugating isoflavones in the glucoside form to the aglycone form. Isoflavones in the aglycone form are absorbed faster and to a greater extent in the human gut than isoflavones in the glucoside form, and isoflavones in the aglycone form are the biologically more active species in the present context. In view hereof it can be understood that administration of a combination of soy proteins, a high, fixed level of isoflavones and a combination of soluble and insoluble fibers may be effective in providing an increased uptake of isoflavones.

Furthermore, again without wishing to be bound by any specific theory, it is presently believed that both soluble dietary fibers (working as nutrients) and insoluble dietary fibers (working as bulking agents), in particular from soybean fibers, more particularly soy cotyledon fibers, provide favorable growth conditions for the microflora in the human gut, which makes the microflora more effective in converting phosphatidyl serine and phosphatidyl ethanolamine into phosphatidyl choline. This capability to decarboxylate phosphatidyl serine into phosphatidyl ethanolamine by the action of pyridoxal phosphate enzymes and further methylate phosphatidyl ethanolamine into phosphatidyl choline has presently only been proven for bacteria. Phosphatidyl choline are absorbed faster and to a greater extent in the human gut than phosphatidyl serine and phosphatidyl ethanolamine, and phosphatidyl choline is the biologically more active species in the present context. In view hereof it can be understood that administration of a combination of soy proteins, a phospholipid source having a high fixed level of phosphoglycerides and a combination of soluble and insoluble fibers may be effective in providing increased levels of phosphatidyl choline from a given phospholipid source and hence provide an increased uptake of phosphatidyl choline from a given phospholipid source.

If present the amount of dietary fibers of the total weight of a composition according to the present invention on a dry basis is preferably more than 4 weight percent, for example at least 5 weight percent, such as at least 6 weight percent, for example at least 7 weight percent, such as at least 8 weight percent, for example at least 9 weight percent, such as at least 10 weight percent, for example at least 11 weight percent, such as at least 12 weight percent, for example at least 13 weight percent, such as at least 14 weight percent, for example at least 15 weight percent, such as at least 16 weight percent, for example at least 17 weight percent, such as at least 18 weight percent, for example at least 19 weight percent, such as at least 20 weight percent, and preferably less than 50 weight percent.

Preferred amounts of dietary fibers as a weight percent of the content of soy protein, shall be in the range of from about 10 to 100 weight percent, and preferred amounts are in the range of from 15 to 90 weight percent, such as from 20 to 80 weight percent, for example 25 weight percent, such as 30 weight percent, for example 33 weight percent, such as 35 weight percent, for example 40 weight percent, such as 50 weight percent, for example 60 weight percent, such as 70 weight percent, for example 75 weight percent.

Accordingly, the weight ratio of soy protein to dietary fibers is from about 1.0 to about 10.0, preferably more than about 1.0, for example about 1.25, such as at least about 1.5, for example at least about 1.75, such as at least about 2.0, for example at least about 2.25, such as at least about 2.5, for example at least about 2.75, such as at least about 3.0, for example at least about 3.25, such as at least about 3.5, for example at least about 3.75, such as at least about 4.0, for example at least about 4.25, such as at least about 4.5, for example at least about 4.75, such as at least about 5.0, for example at least about 5.5, such as at least about 6.0, for example at least about 7.5.

The preferred daily dosage of soybean fibers is from at least 1 g to about 100 g soybean fibers, for example from at least 2 to about 75 g soybean fibers, such as from at least 3 g to about 50 g, for example from at least 4 g to about 40 g, such as from at least 5 to about 30 g, such as from at least 10 g to about 20 g soybean fibers.

In another preferred embodiment, the present invention provides the use of a composition wherein no dietary fibers are present. This composition comprises soy protein, preferably isolated soy protein in an amount of at least 50 weight percent of the total protein content of the composition, said total protein content providing at least 15 percent of the total energy content of the composition, and at least one naturally occurring isoflavone in an amount of more than 0.34 weight percent of the soy protein content of the composition. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament effective in preventing, treating, prophylactically treating, alleviating and/or eliminating a cardiovascular disease in a subject. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament effective in preventing, treating, prophylactically treating, alleviating and/or eliminating arteriosclerosis or a related microvascular or lipoprotein related disease in a subject. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament for treating diabetic subjects, said treatment being effective in lowering serum levels of glucose and/or insulin and/or lipids. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament effective in treating and/or alleviating type 2 diabetes, the metabolic syndrome as defined herein and/or microvascular or lipoprotein related diseases associated therewith in a subject. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament effective in treating subjects diagnosed as having a pulmonary disease, said treatment being effective at least in increasing FEVi of a subject, as measured by forced expiratory volume in the first second of expiration. The present invention also provides the use of such a composition as a medicament and/or in the manufacture of a medicament effective in treating and/or alleviating pulmonary diseases in a subject. The present invention also provides the use of such a composition and/or such a composition for use in treating microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system in a subject. The present invention also provides the use of such a composition and/or such a composition for use in the treatment of diabetic subjects, said treatment being particularly effective in lowering serum levels of glucose and lipids in a subject. The present invention also provides the use of such a composition and/or such a composition for use in the treatment and/or alleviation of a pulmonary disease in a subject, said treatment and/or alleviation resulting in an increased FEVi of a subject, as measured by forced expiratory volume in the first second of expiration. The present invention also provides the use of such a composition and/or such a composition for use in improving physical performance and/or athletic performance or vascularization and conditioning of the skin measured as diminishing and/or smoothening wrinkles.

A composition according to the present invention may optionally comprise a carbohydrate source, flavoring agents, vitamins, minerals, electrolytes, trace elements and other conventional additives. The nutritional composition according to the present invention may in one embodiment also comprise one or more flavoring agents such as cocoa, vanilla, lime, strawberry or soup flavors, such as mushroom, tomato or bouillon and/or sweeteners such as stevia as well as other additives such as xanthan gum. Preferably, the composition contains only natural and plant based substances.

When a carbohydrate source is present in a composition according to the present invention, it is preferably present in an amount of less than 30 weight percent such as less than 25 weight percent of the composition. Preferably, the amount of carbohydrate amounts to at least 5 weight percent, more preferred at least 10 weight percent, and most preferred at least 15 weight percent, of the composition. The preferred carbohydrates for use in a composition according to the present invention are dextrose, fructose and/or maltodextrin, or glucose. Skimmed milk and lecithinated fat reduced cacao are other possible carbohydrate sources. When a composition according to the present invention is for use in the prevention and/or treatment of type 2 diabetes, the metabolic syndrome and associated cardiovascular diseases, lecithinated fat reduced cacao is particularly preferred. Other preferred carbohydrates for use in a composition according to the present invention for use in the prevention and/or treatment of type 2 diabetes, the metabolic syndrome and associated cardiovascular diseases are polydextrose or saccharose, but these should be limited, preferablyusing natural sweeteners like e.g. stevia

Vitamins and minerals may optionally be added to a composition according to the present invention in accordance with the limits laid down by health authorities. A composition according to the present invention may comprise all recommended vitamins and minerals. The vitamins will typically include A, B1, B2, B12, folic acid, niacin, panthotenic acid, biotin, C, D, E and K. The minerals will typically include iron, zinc, iodine, copper, manganese, chromium and selenium. Electrolytes, such as sodium, potassium and chlorides, trace elements and other conventional additives may also be added in recommended amounts.

A presently preferred composition comprises:

Product Composition g/serve

Isolated soy protein 17.2

Soy cotyledon fibre 5.7

Plant Phospholipids incl. Soy Lecithin 3.8

Soy Germ Extract 2.3 The 17.2 g/serve Isolated soy protein is reduced to provide an overall 15 g/serve of soy protein after allowance of soy germ powder just as non-bioactive ingredients are used to flavour, sweeten and improve the mouthfeel of the finished products.

The soy germ extract used in the product enables a higher level of soy isoflavones in the consumed product than what is found in soy protein. Furthermore, soy germ extract provides higher levels of isoflavones in their natural and intact form, as the use of harsh extraction methods through which soy isoflavones may be damaged are entirely avoided. The ratios of individual isoflavones may differ but typical isoflavone content as proportion of total isoflavones (as glycosides) is as follows:

Isoflavone % whole soybean % within soy germ % product

Genistein 50 15 33

Daidzein 40 50 47

Glycitein 10 35 20

Daidzein is the precursor to equol, produced by intestinal bacterial metabolism in around 40-50% of the population and thought to be responsible for several of the cardiovascular benefits of soy. As daidzein is provided at the same or higher levels in the soy germ extract, these cardiovascular benefits are preserved in the product.

The above 15 g of soy protein and 2,3 q of sov germ per serve should comprise a minimum of 3.4mg isoflavones/g of soy protein, thereby guaranteeing a minimum of 50mg isoflavones per serving.

A composition according to the present invention may be used for special dietary use, preferably for lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides in subjects such as hyperlipidemic patients or normocholesterolemic patients suffering from a cardiovascular disease, and/or for lowering serum levels of glucose and/or insulin and/or total cholesterol and/or LDL-cholesterol and/or triglycerides and/or for increasing glucose tolerance and/or insulin sensitivity and/or for preventing, treating and/or alleviating impaired glucose tolerance and/or insulin secretory failure in diabetic subjects and/or for preventing, treating and/or alleviating an arteriosclerotic condition by reducing the influx of lipoproteins and/or cholesterol and/or triglycerides into the endothelium of the arterial wall of a diabetic subject suffering from a cardiovascular disease. For example, from two to three daily meals of ordinary food can be supplemented or replaced by a composition according to the present invention. Hereby, significant reductions in serum levels of cholesterol and/or LDL-cholesterol and/or triglycerides can be obtained, as well as an improvement of serum HDL/LDL-cholesterol ratio and/or an increase in serum HDL-cholesterol levels.

The composition may provide from about 50 to about 250 kcal per serving.

The daily dose of a composition according to the present invention should provide more than 12 g soy protein, such as at least 12.5 g soy protein, for example 13 g soy protein, such as at least 13.5 g soy protein, such as at least 14 g soy protein, for example 15 g soy protein, such as at least 16 g soy protein, for example 17 g soy protein, such as at least 18 g soy protein, for example 19 g soy protein, such as at least 20 g soy protein, for example 21 g soy protein, such as at least 22 g soy protein, for example 23 g soy protein, such as at least 24 g soy protein, for example 25 g soy protein, such as at least 26 g soy protein, for example 27 g soy protein, such as at least 28 g soy protein, for example 29 g soy protein, and preferably at least 30 g soy protein.

The daily dose of a composition according to the present invention should optionally provide more than 5.7 g soy fiber, such as at least 5.8 g soy fibre, for example 5.9 g soy fibre, such as at least 6.0 g soy fibre, for example 6.1 g soy fibre, such as at least 6.2 g soy fibre, for example 6.3 g soy fibre, such as at least 6.4 g soy fibre, for example 6.5 g soy fibre, such as at least 7 g soy fibre, for example 7.5 g soy fibre, such as at least 8 g soy fibre, for example 8.5 g soy fibre, such as at least 9 g soy fibre, for example 9.5 g soy fibre, such as at least 10 g soy fibre, for example 10.5 g soy fibre, such as at least 11 g soy fibre, for example 11.2 g soy fibre, and preferably at least 11.5 g soy fibre.

The daily dose of a composition according to the present invention should optionally provide more than 3.8 g plant phospholipids incl. soy phospholipids, such as at least 3.9 g plant phospholipids incl. soy phospholipids, for example 4.0 g plant phospholipids incl. soy phospholipids, such as at least 4.1 g plant phospholipids incl. soy phospholipids, for example 4.2 g plant phospholipids incl. soy phospholipids, such as at least 4.3 g plant phospholipids incl. soy phospholipids, for example 4.4 g plant phospholipids incl. soy phospholipids, such as at least 4.5 g plant phospholipids incl. soy phospholipids, for example 4.6 g plant phospholipids incl. soy phospholipids, such as at least 4.7 g plant phospholipids incl. soy phospholipids, for example 4.8 g plant phospholipids incl. soy phospholipids, such as at least 4.9 g plant phospholipids incl. soy phospholipids, for example 5.0 g plant phospholipids incl. soy phospholipids, such as at least 5.5 g plant phospholipids incl. soy phospholipids, for example 6.0 g plant phospholipids incl. soy phospholipids, such as at least 6.5 g plant phospholipids incl. soy phospholipids, for example 7.0 g plant phospholipids incl. soy phospholipids, such as at least 7.2 g plant phospholipids incl. soy phospholipids, for example 7.3 g plant phospholipids incl. soy phospholipids, and preferably at least 7.5 g plant phospholipids incl. soy phospholipids.

In a particularly preferred embodiment the daily dose of a composition according to the present invention should provide 2.3 g soy germ extract or more, such as at least 2.4 g soy germ extract, for example 2.5 soy germ extract, such as at least 2.6 g soy germ extract, for example 2.7 soy germ extract, such as at least 2.8 g soy germ extract, for example 2.9 soy germ extract, such as at least 3.0 g soy germ extract, for example 3.1 soy germ extract, such as at least 3.2 g soy germ extract, for example 3.3 soy germ extract, such as at least 3.4 g soy germ extract, for example 3.5 soy germ extract, such as at least 3.6 g soy germ extract, for example 3.7 soy germ extract, such as at least 3.8 g soy germ extract, for example 3.9 soy germ extract, such as at least 4.0 g soy germ extract, for example 4.1 soy germ extract, such as at least 4.2 g soy germ extract, for example 4.25 soy germ extract, such as at least 4.3 g soy germ extract, for example 4.35 soy germ extract, such as at least 4.4 g soy germ extract, for example 4.45 soy germ extract, such as at least 4.5 g soy germ extract, for example 4.55 soy germ extract, and preferably at least 4.6 g soy germ extract.

Whether in the form of a soy protein source with a high concentration of isoflavones and/or in the form of soy germ the daily dose of a composition according to the present invention should provide more than 50 mg isoflavones (genistein, daidzein, glycitein), such as at least 55 mg isoflavones, for example 60 mg isoflavones, such as at least 70 mg isoflavones, for example 75 mg isoflavones, such as at least 80 mg isoflavones, for example 85 mg isoflavones, such as at least 90 mg isoflavones, for example 91 mg isoflavones, such as at least 92 mg isoflavones, for example 93 mg isoflavones, such as at least 94 mg isoflavones, for example 95 mg isoflavones, such as at least 96 mg isoflavones, for example 97 mg isoflavones, such as at least 98 mg isoflavones, for example 99 mg isoflavones, and preferably at least 100 mg isoflavones. Preferably the daily dose of a composition according to the present invention should provide at least 30 g soy protein, 11 ,5 g soy fiber, 7,5 g plant phospholipids incl. soy phospholipids, and a minimum of 100 mg isoflavones (genistein, daidzein, glycitein), e.g. in the form of 4.6 g soy germ extract,.

The daily dose of a composition according to the present invention may comprise an energy content of from 400 to 800, in particular from 450 to 800 kcal/day, which is considered to be a very low calorie diet (VLCD), or it may comprise an energy content of from 800 to 1200 kcal/day, which is considered to be a low-calorie diet (LCD). In another medical embodiment of the present invention, the energy content may correspond to the energy requirement of a normal person.

The present invention also provides a composition according to the invention in the form of a micronutrient. In this connection a micronutrient is a nutritional supplement and/or a pharmacological composition and/or a medicament comprising i) a synthetic phytoestrogen-like compound capable of binding to an estrogen receptor or an estrogen-like receptor, and/or ii) a naturally occurring, plant-extractable compound in an amount, on a weight per weight basis, in excess of the amount of said compound, when it is present in a natural host such as a plant cell from which the compound can be extracted or isolated, iii) soy peptides obtainable from a partial hydrolysis of soy protein and iv) soy lecithin.

The naturally occurring, plant-extractable compound is preferably but not limited to compounds capable of binding to an estrogen receptor, an estrogen-like receptor, a beta-2-adrenergic receptor or a receptor belonging to the class of beta-2-adrenergic receptors. When the naturally occurring compounds are isolated from plants such as soybeans, they may be selected from the group at least containing phytoestrogens such as soybean phytoestrogens such as soybean isoflavones, soy protein or fragments thereof, e.g. peptides or amino acid sequences, soybean fibers, lecithin, linolenic acid, an antioxidant, a saponin, a lignan, a protease inhibitor, a trypsin inhibitor, and a tyrosine kinase inhibitor. Additional constituents of the micronutrient may preferably be selected among a DNA topoisomerase inhibitor, a ribosome kinase inhibitor, a growth control factor such as e.g. epidermal growth factor, transforming growth factor alpha, platelet derived growth factor, and preferably any growth control factor controllable by a tyrosine kinase activity. The micronutrient may also comprise ormeloxifene and/or levormeloxifene as described by among others Holm et al. (1997) in Arteriosclerosis, Thrombosis, and Vascular Biology 17 (10), 2264 - 2272, and in Clinical Investigation, 100 (4), 821 - 828. When the naturally occurring compound is an isoflavone, the isoflavone may have been deconjugated to the aglycone form either biologically or in vitro prior to the incorporation in the micronutrient.

In one particularly preferred embodiment the present invention provides a composition or a micronutrient according to the present invention in combination with a functional food ingredient comprising a sterol, preferably an ingredient selected from the group comprising a stand ester, a tocotrienol, a mevinolin, and a phytosterol compound such as e.g. campesterol, sitosterol or stigmasterol, or a combination thereof.

According to one preferred embodiment, a composition or a micronutrient according to the present invention is for use as a functional food ingredient. A composition or a micronutrient according to the present invention may also be administered as a probe or by intravenous administration, or in tablet or capsule form. The present invention also provides a pharmaceutical preparation comprising the a composition or a micronutrient according to the present invention, use of the a composition or a micronutrient according to the present invention in therapy and/or a diagnostic method performed on the human or animal body, use of a composition or a micronutrient according to the present invention in the manufacture of a medicament, use of a composition or a micronutrient according to the present invention in the manufacture of a medicament for treating a subject suffering from cardiovascular diseases, use of a composition or a micronutrient according to the present invention in the manufacture of a medicament for treating a subject suffering from type 2 diabetes, the metabolic syndrome and/or cardiovascular diseases associated therewith in a diabetic subject and use of a composition or a micronutrient according to the present invention in the manufacture of a medicament for treating a subject suffering from pulmonary diseases.

The micronutrient is particularly useful in preventing, treating, prophylactically treating and/or alleviating hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis and/or related cardiovascular diseases and in preventing and/or treating type 2 diabetes, the metabolic syndrome and/or cardiovascular diseases associated therewith in a diabetic subject and in preventing, treating, prophylactically treating and/or alleviating a pulmonary disease such as e.g. a disease selected from the group comprising inflammation of the airways, bronchoconstriction, bronchitis, asthma, and small airways diseases.

The micronutrient is also useful for improving the physical performance and/or athletic performance as measured in watts of a human or animal subject, or for improving vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles, of a human or animal subject.

In one embodiment the present invention provides a composition according to the present invention for use as a medicament or as a dietary preparation. A composition according to the present invention for use as a medicament or as a dietary preparation may preferably be used in preventing, treating, prophylactically treating and/or alleviating cardiovascular diseases such as e.g. a disease selected from the group comprising hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, myocardial infarction, and hypertension, in a subject, preferably for use in preventing, treating, prophylactically treating and/or alleviating arteriosclerosis and/or atherosclerosis in a subject . A composition according to the present invention for use as a medicament or as a dietary preparation may also preferably be used in preventing, treating, alleviating and/or eliminating type 2 diabetes. A composition according to the present invention for use as a medicament or as a dietary preparation may also preferably be used in preventing, treating, alleviating and/or eliminating a cardiovascular disease, such as e.g. hypercholesterolemia, hypertriglyceridemia, hypertension, hyperglycemia, hyperinsulinemia, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, hypertension, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, and myocardial infarction, in a diabetic subject. A composition according to the present invention for use as a medicament or as a dietary preparation may also preferably be used for preventing and/or treating pulmonary diseases, such as preferably a disease selected from the group comprising inflammation of the airways, bronchoconstriction, bronchitis, asthma, and small airways diseases, in a subject. A composition according to the present invention for use as a medicament or as a dietary preparation may also preferably be used for improving the physical performance and/or athletic performance as measured in watts of a human or animal subject, or for improving vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles, of a human or animal subject.

The present invention also provides the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing, treating, prophylactically treating and/or alleviating cardiovascular diseases such as e.g. a disease selected from the group comprising hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, myocardial infarction, and hypertension, particularly a disease selected from the group comprising arteriosclerosis and atherosclerosis, in a subject. The present invention also provides the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing, treating and/or alleviating type 2 diabetes and/or the metabolic syndrome in a subject and/or a cardiovascular disease in a diabetic subject. The present invention also provides the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing, treating, prophylactically treating and/or alleviating pulmonary diseases such as e.g. a disease selected from the group comprising inflammation of the airways, bronchoconstriction, bronchitis, asthma, and small airways diseases, in a subject.

The composition according to the present invention is effective in normalizing face color and capillary blood flow in subjects, otherwise having a gusty and pale face complexion, within 4 weeks, such as within 3 weeks, for example within 1-2 weeks.

The composition according to the present invention is effective in reducing otherwise increased blood pressure (caused by restricted peripheral blood flow) within 4 weeks, such as within 3 weeks, for example within 1-2 weeks.

The composition according to the present invention is effective in reducing the need for anti-hypertensive medication to half the original dosing within 6 months, and one third the original dosing within 9 months, such as to half the original dosing within 5 months, and one third the original dosing within 8 months, for example to half the original dosing within 4 months, and one third the original dosing within 7 months, such as to half the original dosing within 3 months, and one third the original dosing within 6 months.

The composition according to the present invention is effective in reducing lipid levels, and signs of atherosclerosis and arterial fatty streak formation, in individuals otherwise having a family history of atherosclerosis, and elevated LDL and total cholesterol levels.

The composition according to the present invention is effective in preserving muscle mass and improving physical performance during metabolic stress situations, just like it is effective in enabling a faster recovery after heavy exercise and gives rise to a reduced tenderness in muscles.

The composition according to the present invention is effective in significantly improving physical performance in the form of more than 5% increased physical output (measured as Watts in bicycling tests for 45 min at 130 pulse rate per minute), for example more than 10% increased physical output (measured as Watts in bicycling tests for 45 min at 130 pulse rate per minute), such as more than 15% increased physical output (measured as Watts in bicycling tests for 45 min at 130 pulse rate per minute), for example more than 20% increased physical output (measured as Watts in bicycling tests for 45 min at 130 pulse rate per minute).

The composition according to the present invention is effective in lowering levels of cholesterol in normocholesterolemic patients by at least 2%, for example at least 5%, such as at least 8%, for example at least 10%, such as at least 12%, for example at least 14%, such as at least 16%, for example at least 18%, such as at least 20%, for example at least 25%, such as at least 30%. The composition according to the present invention is effective in lowering levels of triglycerides in normocholesterolemic patients by at least 10%, such as at least 12%, for example at least 14%, such as at least 16%, for example at least 18%, such as at least 20%, for example at least 25%, such as at least 30%.

The composition according to the present invention is effective in lowering levels of total- and LDL-cholesterol in mildly hypercholesterolemic patients by at least 3%, for example at least 5%, such as at least 8%, for example at least 10%, such as at least 12%, for example at least 15%, such as at least 20%, for example at least 25%, such as at least 30%, for example at least 35%, such as at least 40%, for example at least 45%. The composition according to the present invention is effective in lowering levels of triglycerides in mildly hypercholesterolemic patients by at least 15%, such as at least 20%, for example at least 25%, such as at least 30%, for example at least 35%, such as at least 40%, for example at least 45%.

The composition according to the present invention is effective in lowering levels of total- and LDL-cholesterol in severely hypercholesterolemic patients by at least 3%, for example at least 5%, such as at least 8%, for example at least 10%, such as at least 12%, for example at least 15%, such as at least 20%, for example at least 25%, such as at least 30%, for example at least 35%, such as at least 40%, for example at least 45%, such as at least 50%, for example at least 55%, such as at least 60%. The composition according to the present invention is effective in lowering levels of triglycerides in severely hypercholesterolemic patients by at least 20%, for example at least 25%, such as at least 30%, for example at least 35%, such as at least 40%, for example at least 45%, such as at least 50%, for example at least 55%, such as at least 60%.

A composition according to the present invention for use as a medicament and/or the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing, treating, prophylactically treating and/or alleviating cardiovascular diseases in a subject may be effective in reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or causing dilation of blood vessels and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or lowering serum levels of total cholesterol and/or LDL-cholesterol and/or homocysteine and/or triglycerides and/or increasing the serum HDL/LDL-cholesterol ratio and/or increasing serum levels of HDL-cholesterol in a subject and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or reducing or eliminating the risk of a subject contracting angina pectoris and/or reducing or eliminating the risk of a subject contracting a myocardial infarction.

A composition according to the present invention for use as a medicament and/or the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing and/or treating diabetes and/or the metabolic syndrome and/or a cardiovascular disease associated therewith in a subject may be effective in i) lowering serum glucose levels and/or ii) reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or causing dilation of blood vessels and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or iii) lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or homocysteine and/or increasing the serum HDL/LDL-cholesterol ratio and/or serum HDL-cholesterol levels and/or iv) increasing glucose tolerance and/or insulin sensitivity and/or v) alleviating impaired glucose tolerance and/or insulin secretory failure and/or improving insulin secretion and/or vi) preventing, treating, alleviating, and/or eliminating cardiovascular diseases, such as e.g. hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, myocardial infarction, hypertension, hyperglycemia, and hyperinsulinemia, in a diabetic subject and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or reducing or eliminating the risk of a diabetic subject contracting angina pectoris and/or reducing or eliminating the risk of a diabetic subject contracting a myocardial infarction and/or in treating a procoagulant state and/or an increased activity of clotting factors, insulin resistance, glycosidation and/or oxidation and/or chemical modification of lipoproteins, as well as impaired glucose tolerance.

A composition according to the present invention for use as a medicament and/or the use of a composition according to the present invention as a medicament and/or in the manufacture of a medicament for preventing, treating, prophylactically treating and/or alleviating pulmonary diseases may be effective in i) preventing, treating, prophylactically treating and/or alleviating asthma and/or ii) reducing and/or eliminating mucus hypersecretion and/or dyspnea in a subject suffering from asthma and/or iii) increasing FEVi of a subject as measured by forced expiratory volume in the first second of expiration and/or iv) preventing, treating, prophylactically treating, alleviating and/or reducing inflammation of the airways and/or v) preventing, treating, prophylactically treating and/or alleviating bronchoconstriction.

In another embodiment the present invention provides the use of a composition according to the present invention in the treatment of cardiovascular diseases in the human or animal body in an amount effective in lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or homocysteine and/or increasing the serum HDL/LDL-cholesterol ratio and/or serum HDL-cholesterol levels and/or reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or reducing or eliminating the risk of a subject contracting angina pectoris and/or reducing or eliminating the risk of a subject contracting a myocardial infarction, and/or alleviating the clinical condition of patients contracting a myocardial infection. The cardiovascular disease is preferably a cardiovascular disease selected from the group comprising hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, myocardial infarction, and hypertension and more preferred selected from arteriosclerosis and atherosclerosis.

In another embodiment the present invention provides the use of a composition according to the present invention in the treatment of type 2 diabetes and/or the metabolic syndrome in an amount effective in lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or glucose and/or increasing serum levels of H LDL-cholesterol and/or homocysteine and/or reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or improving glucose tolerance and/or increasing insulin sensitivity and/or alleviating impaired glucose tolerance and/or improving insulin secretion and/or reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or preventing, reducing or eliminating the risk of a subject contracting angina pectoris and/or preventing, reducing or eliminating the risk of a subject contracting a myocardial infarction and/or preventing, treating, prophylactically treating, alleviating and/or eliminating hypertension and/or hyperglycemia and/or hyperinsulinemia and/or hypercholesterolemia and/or hypertriglyceridemia and/or arteriosclerosis and/or atherosclerosis and/or arteriolosclerosis in a diabetic subject. In another embodiment the present invention provides the use of a composition according to the present invention in the treatment of a pulmonary disease in a human or animal body, preferably a disease selected from the group comprising inflammation of the airways, bronchoconstriction, bronchitis, asthma, and small airways diseases, in an amount effective in preventing, treating, prophylactically treating and/or alleviating inflammation of the airways and/or bronchoconstriction and/or bronchitis and/or small airways diseases and/or asthma and/or reducing and/or eliminating mucus hypersecretion and/or dyspnea in a subject suffering from asthma and/or increasing FEVi of a subject as measured by forced expiratory volume in the first second of expiration.

The present invention also provides a method of preventing, treating, prophylactically treating and/or alleviating by therapy a cardiovascular disease in the human or animal body such as an arteriosclerotic condition of a human or animal body, said method comprising administration of a composition according to the present invention in an amount effective in lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or homocysteine and/or increasing the serum HDL/LDL- cholesterol ratio and/or serum HDL-cholesterol levels and/or reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or reducing or eliminating the risk of a subject contracting angina pectoris and/or reducing or eliminating the risk of a subject contracting a myocardial infarction, and/or alleviating the clinical condition of patients contracting a myocardial infection. The cardiovascular disease is preferably a cardiovascular disease selected from the group comprising hypercholesterolemia, hypertriglyceridemia, other hyperlipidemias, arteriosclerosis, atherosclerosis, arteriolosclerosis, capillary sclerosis, obesity/overweight, Alzheimer, Dementia, coronary heart disease, angina pectoris, thrombosis, myocardial infarction, and hypertension and more preferred selected from arteriosclerosis and atherosclerosis.

The present invention also provides a method of preventing and/or treating by therapy type 2 diabetes and/or the metabolic syndrome in a human or animal body, said method comprising administration to said human or animal body of a composition according to the present invention in an amount effective in lowering serum levels of total cholesterol and/or LDL-cholesterol and/or triglycerides and/or glucose and/or increasing serum levels of H LDL-cholesterol and/or homocysteine and/or reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or improving glucose tolerance and/or increasing insulin sensitivity and/or alleviating impaired glucose tolerance and/or improving insulin secretion and/or reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation and/or preventing, reducing or eliminating complicated lesion formation and/or preventing, reducing or eliminating the risk of a subject contracting angina pectoris and/or preventing, reducing or eliminating the risk of a subject contracting a myocardial infarction and/or preventing, treating, prophylactically treating, alleviating and/or eliminating hypertension and/or hyperglycemia and/or hyperinsulinemia and/or hypercholesterolemia and/or hypertriglyceridemia and/or arteriosclerosis and/or atherosclerosis and/or arteriolosclerosis in a diabetic subject.

The present invention also provides a method of preventing, treating, prophylactically treating and/or alleviating by therapy a pulmonary disease in a human or animal body, preferably a disease selected from the group comprising inflammation of the airways, bronchoconstriction, bronchitis, asthma, and small airways diseases, said method comprising administration to said human or animal body of a composition according to the present invention in an amount effective in preventing, treating, prophylactically treating and/or alleviating inflammation of the airways and/or bronchoconstriction and/or bronchitis and/or asthma and/or small airways diseases and/or reducing and/or eliminating mucus hypersecretion and/or dyspnea in a subject suffering from asthma and/or increasing FEVi of a subject as measured by forced expiratory volume in the first second of expiration.

The period of treatment is preferably in the range of from 1 to 12 months or more, such as from 2 weeks to 9 months, for example from 3 weeks to 6 months, such as from 4 weeks to 4 months, such as from 6 weeks to 3 months. However, the period of treatment shall not be limited to these periods and may e.g. be longer than 12 months, such as e.g. a lifelong treatment in order to prevent cardiovascular diseases or in order to prevent and/or alleviate type 2 diabetes and/or cardiovascular diseases such as microvascular or lipoprotein related diseases in connection therewith or in order to prevent pulmonary diseases. In one embodiment the present invention provides a pharmaceutical preparation comprising a composition according to the present invention. The pharmaceutical preparation can be prepared in any way known to the skilled person.

In another embodiment the present invention provides the use of a composition according to the present invention as a nutritional preparation and/or in the manufacture of a nutritional preparation for lowering serum levels of glucose and/or total cholesterol and/or LDL-cholesterol and/or triglycerides and/or homocysteine and/or increasing the serum HDL/LDL-cholesterol ratio and/or serum levels of HDL- cholesterol in a subject, including a diabetic subject, and/or for alleviating a pulmonary condition such as e.g. asthma. The nutritional preparation may take any form, which is suitable for human or animal consumption. In one preferred embodiment, the composition is a powdery mixture, which is suspendable, dispersible or emulsifiable in a liquid for human or animal consumption. The liquid is preferably a water-containing liquid such as e.g. water, coffee, tea or juice, including fruit juice. For such a purpose, the composition may be packed in a package intended for covering part of or the total nutritional requirement for a defined period of time, such as two or more compositions a day over a period of e.g. three days or a week. The present invention also provides the nutritional preparation in the form of a dietary supplement.

The nutritional preparation in one embodiment of the present invention is preferably a nutraceutical product, functional food or drink, i.e. a readily obtainable edible or drinkable substance that is supplemented with a composition according to the present invention to provide a medical or pharmaceutical effect. Accordingly, the present invention provides a composition according to the present invention for use as a functional food ingredient.

In another embodiment, a composition according to the present invention is a liquid nutritional preparation in a water-containing liquid, in which the solid ingredients are suspended, dispersed or emulgated in an amount from 10 to 25 weight percent. When the liquid nutritional preparation is intended for drinking, it will usually comprise a flavoring agent as discussed above. However, the liquid nutritional preparation may also be used for probe administration.

In another embodiment, the present invention relates to the use of a composition according to the present invention as a partial or total diet for an overweight subject, an overweight subject suffering from an arteriosclerotic condition or an overweight subject suffering from a diabetic condition. Obesity is believed to be one of the major causes of diabetes including type 2 diabetes. Overweight subjects, including overweight diabetic subjects, often have increased serum cholesterol levels and increased triglyceride levels and are therefore more likely to develop cardiovascular diseases. However, the present invention is not limited to treating subjects with an increased risk of contracting a cardiovascular disease, i.e. subjects likely to have increased serum levels of cholesterol and/or triglycerides, or to treating obese diabetic subjects with an increased risk of contracting a cardiovascular disease, i.e. obese diabetic subjects likely to have increased serum levels of cholesterol and/or triglycerides. A composition according to the present invention also has substantial serum cholesterol, serum LDL-cholesterol and serum triglyceride lowering effects in subjects having a more normal lipid profile and in diabetic subjects that do not also suffer from overweight. The medical use of a composition according to the present invention is not limited to overweight or obese subjects, including diabetic subjects, but may be used for normal weight subjects having increased serum levels of cholesterol and/or LDL-cholesterol and/or triglycerides or for subjects with a cardiovascular condition such as e.g. microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system or a related condition who have normal serum levels of cholesterol and/or LDL-cholesterol and/or triglycerides. Such increased serum levels of cholesterol and/or LDL-cholesterol and/or triglycerides may be caused by intake of a diet rich in fats or it may be genetically related.

For the purpose of the present invention, subjects having an initial total serum cholesterol level of 5.7 mmol/l or below are considered to have a normal or hypocholesterolemic level, whereas subjects having a total serum cholesterol level above 5.7 mmol/l are considered to be hypercholesterolemic. Accordingly, by treating normocholesterolemic subjects, it is possible to prevent the development of cardiovascular diseases arising from serum cholesterol levels below a concentration of 5.7 mmol/l in subjects, including diabetic subjects, particularly sensitive to developing e.g. microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system, or prevent further development of cardiovascular diseases in patients, including diabetic patients, with previous cardiovascular events.

By treating hypercholesterolemic subjects, it is possible to prevent the development of cardiovascular diseases arising from serum cholesterol levels above a concentration of 5.7 mmol/l in subjects sensitive to developing e.g. microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system under such conditions.

More particularly, subjects having a total serum cholesterol level of from 5.7 mmol/l to 7.9 mmol/l are considered to be mildly hypercholesterolemic. Accordingly, by treating these hypercholesterolemic subjects, it is possible to prevent the development of cardiovascular diseases arising from serum cholesterol levels of from 5.7 to 7.9 mmol/l. Subjects having a total serum cholesterol level of more than 7.9 mmol/l are considered to be severely hypercholesterolemic. Accordingly, by treating these hypercholesterolemic subjects, it is possible to prevent the development of cardiovascular diseases arising from serum cholesterol levels of more than 7.9 mmol/l.

It has also been shown that a composition according to this invention has a potentiat ing effect to the effect of medications such as e.g. statins and/or niacin. By combining a composition according to the present invention with e.g. statins, such as HMG-CoA- reductase-inhibitors, niacin, bile acid resins, fibrates, nicotinic acid derivatives, oat products, such as oat meal, rye products, such as rye meal and various fish oil con centrates with a high content of w-3-fatty acids, it is possible to achieve a further 5 to 15% reduction in serum levels of total cholesterol and/or LDL-cholesterol and/or tri glycerides. The present invention also provides a composition according to the present invention in combination with a statin, preferably an HMG-CoA-reductase-inhibitor, niacin, bile acid resins, fibrates, oat products, rye products, nicotinic acid derivatives and various fish oil concentrates with a high content of w-3-fatty acids.

EXAMPLES

EXAMPLE 1

The product used in this example comprises 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein).

The soy germ extract used in the product enables a higher level of soy isoflavones in the consumed product than what is found in soy protein alone. Furthermore, soy germ extract provides higher levels of isoflavones in their natural and intact form (as the use of harsh extraction methods through which soy isoflavones may be damaged are entirely avoided). The ratios of individual isoflavones may differ but typical isoflavone content as proportion of total isoflavones (as glycosides) is as follows:

Isoflavone % whole soybean % within soy germ % product Genistein 50 15 33

Daidzein 40 50 47

Glycitein 10 35 20

The above 15g of isolated soy protein per serve comprised a minimum of 3.4mg isoflavones/g of soy protein, thereby guaranteeing a minimum of 50 mg isoflavones per serving.

In heart patients otherwise having a gusty and pale face complexion, intake of this product twice daily lead to a normalized face color and capillary blood flow within 1-2 weeks. Furthermore, otherwise increased blood pressures (caused by restricted peripheral blood flow) were significantly reduced:

Example A: Male, 61 years old, with high blood pressure and pale face complexion, diagnosed with atherosclerosis and with stents in coronary and kidney arteries. Intake of a product providing 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones per serving (genistein, daidzein, glycitein), taken twice daily, lead to normalized reddish face color, as well as normalized blood pressure and substantially lower intake of anti hypertensive medications, which was reduced to half intake after 3 months, and one third after 6 months.

Example B: Male, 55 years old, family history of atherosclerosis, and substantially elevated LDL and total cholesterol levels. Intake of product providing 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein) twice daily for > 20 years, substantially reduced lipid levels, and MRI using contrast agent revealed no sign of atherosclerosis or arterial fatty streak formation, which normally starts in mid 20s.

Example C: Male 48 years. Intake of product providing 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein) 4-5 times daily lead to significant reduction of blood pressure from 230/130 to 133/82 in just 3 days.

Example D: Male 60 years, diagnosed with severe heart failure and hypertension, diabetic for past 10 years with nerve damages to legs. Intake of product providing 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein) 2 times daily lead to significant reduction of blood pressure within few days, and normalized heart function within 2 weeks.

Example E: Male, 75 years of age, diagnosed with atherosclerosis in coronary arteries, of which two areas in the coronary arteries were so blocked that they gave rise to serious angina pectoris (chest pains). The one blocked area of the coronary artery was treated with a stent, and the person put on a waiting list for a coronary bypass operation for the second blocked coronary artery. Intake of product providing 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein) twice daily resulted in the disappearance of angina pectoris according to regular controls with health personal, just like the person’s initial white face complexion quickly disappeared, and was replaced by a normal red colour of the face and. When coronary arteries were checked during hospitalization after 12 months of intake of the above- mentioned product, there was no longer a need for a coronary bypass operation since the atherosclerosis and blockage previously seen had almost disappeared.

EXAMPLE 2

The product used in this example comprises 15 g soy protein, 5.7 g soy fiber, 3.8 g plant phospholipids incl. soy phospholipids, 2.3 g soy germ extract, and a minimum of 50 mg isoflavones (genistein, daidzein, glycitein) per serving, taken twice daily.

The soy germ extract used in the product enables a higher level of soy isoflavones in the consumed product than what is found in soy protein. Furthermore, soy germ extract provides higher levels of isoflavones in their natural and intact form (as the use of harsh extraction methods through which soy isoflavones may be damaged are entirely avoided). The ratios of individual isoflavones may differ but typical isoflavone content as proportion of total isoflavones (as glycosides) is a s follows:

Isoflavone % whole soybean % within soy germ % product

Genistein 50 15 33

Daidzein 40 50 47

Glycitein 10 35 20

The above product was consumed twice daily by a group of athletes during the 9 month-long Volvo Ocean Race, one of the world^'s most enduring sports competition

Controlled measurements enabled the conclusion that the intake twice daily of the product preserved muscle mass and improved physical performance during metabolic stress situations in top athletes, just like it enabled faster recovery after heavy exercise and reduced tenderness in muscles.

Tests showed a significantly improved physical performance by subjects consuming the product twice daily, in the form of a 20-30% increased physical output (measured as Watts in bicycling tests for 45 min at 130 pulse rate per minute).

EXAMPLE 3

A patient hospitalized with life threatening high blood pressure experienced a weight loss of approx. 25 kg after three months, following administration of a composition according to the present invention, i.e. approx reduction of 175.000 kcal, blood pressure reduced from 230/130 on strong anti-hypertensive medications, to 133/82 without medications.

This highly significant blood pressure reduction happened within the first 3 days administration of a composition according to the present invention, i.e. much faster than the weight loss (just like the rapid disappearance of angina pectoris in heart patients), and is speculated to have been caused by the known vasodilation mechanisms of arginine and isoflavones, respectively. The normalization of blood pressure was continued to be observed also following 3 months of administration of a composition according to the present invention, and after anti-hypertensive medications had been seponated. In a similar manner Blood sugar levels normalized in just a few days, which allowed for the seponation of type 2 diabetes medications.

EXAMPLE 4

A patient experienced a 22 kg weight loss after 11 weeks of administration of a composition according to the present invention 5 times per day, approx every 3 hours. Blood pressure and blood sugar levels were normalised, allowing for seponation of anti-hypertensive and type 2 diabetes medicines.

This highly significant blood pressure reduction happened within the first 3 days of administration of a composition according to the present invention and is speculated to have been caused by the known vasodilation mechanisms of arginine and isoflavones, respectively.

EXAMPLE 5

A soy based dietary supplement composition according to the present invention was compared with casein in relation to its ability to attenuate aortic cholesterol accumulation in rabbits clamped at the same hypercholesterolemic level.

A composition according to the present invention in the form of a dietary supplement containing isolated soy protein (supro soy) with standardized high levels of isoflavones, cotyledon soy fibre and soy phospholipids had in previous experiments in comparison with casein shown a strong dose dependent plasma cholesterol lowering effect in rabbits fed cholesterol enriched diet. The present study was aimed at investigating whether a composition according to the present invention would in addition have a direct effect on cholesterol accumulation in the arterial wall not mediated by plasma cholesterol lowering.

Four groups each with 20 male rabbits were fed a diet enriched with 80% of the composition according to the present invention, 40% of the composition according to the present invention, 40% casein and 40% casein. The latter group received estrogen injections intramuscularly biweekly and was included as a positive control, since estrogen is known to have a direct antiatherogenic effect on the arterial wall in cholesterol fed rabbits. All rabbits were maintained at the same elevated plasma cholesterol level of about 20 mmol/l for 75 days by individualized cholesterol feeding procedure.

The rabbits receiving 80% of the composition according to the present invention and the rabbits receiving estrogen required in average twice and three times as much dietary cholesterol respectively as the rabbits receiving only casein. As shown in figure 7, the aortic cholesterol concentrations in nmol/mg (SE) tissue were 6.5 (0.7), 7.8 (0.9), 10.2 (1.0) and 7.5 (0.6) in the 80% of the composition according to the present invention, 40% of the composition according to the present invention, the casein group and in the estrogen group, respectively with a significant difference (p<0.01) between the rabbits receiving 80% of the composition according to the present invention and casein + estrogen compared with the rabbits receiving only casein.

The composition according to the present invention thus reduces plasma cholesterol and in addition it attenuates cholesterol accumulation in the arterial wall beyond that which can be ascribed to its plasma cholesterol lowering effect. The plasma cholesterol clamped rabbit may be useful for identification of the ingredient(s) in soy products responsible for the direct effect on the arterial wall. The present findings also suggest that investigation of soy products in humans should include its effect on endothelial function.

EXAMPLE 6

This example concerns a randomized, placebo-controlled, double-blind study of a duration of an 8-week treatment period. Initial serum cholesterol was 5.8 - 7.9 mmol/l. The test material was the composition according to the present invention, compared to isolated soy protein and caseinate (placebo).

The trial subject information was as follows:

Subjects included: 121

Subjects not evaluated: 5

Subjects evaluated: 116* (62 female, 54 male)

Premature terminations: 2

Completed the study: 114 *Distributed as: Verum 1 = 39

Verum 2 = 39 Placebo = 38

Changes in LDL cholesterol (LDL-C) from baseline after 8 weeks are shown in Figure 1.

Changes in total cholesterol (Total-C) from baseline after 8 weeks are shown in Figure 2.

Changes in apolipoprotein B from baseline after 8 weeks are shown in Figure 3. Changes in triglycerides from baseline after 8 weeks are shown in Figure 4.

Lipid lowering effects of the composition according to the present invention and isolated soy protein (ISP) are shown in Figure 5.

Superior lipid lowering effects of the composition according to the present invention compared to isolated soy protein (ISP) are shown in Figure 6.

The conclusion was that the placebo-controlled clinical study, performed under Good Clinical Practice, demonstrates that the composition according to the present invention is more than twice as effective in lowering cholesterol compared to isolated soy protein alone.

Claims

1. A composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising:

(a) a soy protein source, which provides for at least 12.5 g of soy protein and at least 50 weight percent of the total protein content of the composition,

(b) a source of naturally occurring soy isoflavones, in the form of a soy protein source with a high concentration of naturally occurring isoflavones and/or a soy germ extract, said source of naturally occurring isoflavones providing naturally occurring isoflavones in an amount of minimum 50 mg and sufficient to ensure that the total content of isoflavones of the composition amounts to at least 0.34 weight percent but less than 2.50 weight percent of the soy protein content of the composition, for use in preventing, treating and/or alleviating any of the following in a human subject: microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system, such as, arteriolosclerosis atherosclerosis, Monckeberg’s mediasclerosis arteriolosclerosis, capillary sclerosis, atherosclerotic cardiovascular disease and/or cerebrovascular small vessel disease (CSVD), menopausal symptoms or premenstrual syndrome, high blood pressure, elevated blood lipids and blood sugar levels, hormonal prostate, breast or colon cancers, inflammation of e.g. joints and tendons, pulmonary diseases such as airway inflammation, asthma, bronchitis and small airways diseases.

2. A composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising:

(a) a soy protein source, which provides for at least 12.5 g of soy protein and at least 50 weight percent of the total protein content of the composition, (b) a source of naturally occurring soy isoflavones, in the form of a soy protein source with a high concentration of naturally occurring isoflavones and/or a soy germ extract, said source of naturally occurring isoflavones providing naturally occurring isoflavones in an amount of minimum 50 mg and sufficient to ensure that the total content of isoflavones of the composition amounts to at least 0.34 weight percent but less than 2.50 weight percent of the soy protein content of the composition, for use in improving in a human or animal subject: physical performance and/or athletic performance by a human or animal by at least 5% as measured in watts in bicycling tests for 45 min at 130 pulse rate per minute, or vascularization and conditioning of the skin measured as a diminishing and/or smoothening of wrinkles.

3. A composition according to claims 1 or 2 further comprising:

(c) a phospholipid source, such as a soy phospholipid source, in an amount sufficient to ensure that the total content of plant phospholipids of the composition amounts to at least 15 weight percent of the soy protein content of the composition, and

(d) a fibre source, such as a soy fibre source, such as a soy cotyledon fibre source in an amount sufficient to ensure that the total content of fibres, such as soy fibres, such as soy cotyledon fibres of the composition amounts to at least 25 weight percent of the soy protein content of the composition.

4. A composition according to any of claims 1 to 3 wherein said naturally occurring isoflavones are selected from the group comprising genistein, daidzein, and glycitein.

5. A composition according to claim 4 wherein some or all of the naturally occurring isoflavones are present in the aglycone form.

6. A composition according to any of claims 3 to 5 wherein said soy protein source is isolated soy protein or soy protein concentrate and wherein said phospholipid source comprises soy lecithin.

7. A composition according to any of claims 1 to 6 wherein the ratio between the amount of arginine in the soy protein source of the composition and the amount of lysine in the soy protein source of the composition is at least 2.

8. A composition according to any of claims 1 or 3 to 7 for use in preventing, treating and/or alleviating in a human or animal subject microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system by reducing the influx of lipoproteins such as LDL cholesterol and/or triglycerides through the endothelium into the arterial wall and/or causing dilation of blood vessels and/or reducing restriction of blood vessels and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation in blood vessels.

9. A composition according to claim 8 where said microvascular or lipoprotein related diseases are selected from the group comprising arteriosclerosis, atherosclerosis, arteriolosclerosis and capillary sclerosis.

10. A composition according to any of claims 1 or 3 to 9 for use in preventing, treating and/or alleviating microvascular or lipoprotein related diseases in patients with hypertension, obesity/overweight, type 2 diabetes, Alzheimer disease and/or Dementia.

11. A composition according to any of claims 1 to 10 for use as a partial or total diet in an amount resulting in a consumption of at least 100 mg of isoflavones on a daily basis.

12. A composition according to any of claims 1 to 11 for use as a partial or total diet for

- a diabetic human or animal subject, or

- an overweight human or animal subject.

13. A method of preventing, treating, prophylactically treating and/or alleviating by therapy any of the following in a human or animal subject: microvascular or lipoprotein related diseases of the heart, vascular and/or central nervous system, such as, arteriolosclerosis atherosclerosis,

Monckeberg’s mediasclerosis arteriolosclerosis, capillary sclerosis, atherosclerotic cardiovascular disease and/or cerebrovascular small vessel disease (CSVD) menopausal symptoms or premenstrual syndrome high blood pressure, elevated blood lipids and/or elevated blood sugar levels, hormonal prostate, breast or colon cancers, inflammation of e.g. joints and tendons, pulmonary diseases such as airway inflammation, asthma, bronchitis and small airways diseases, said method comprising administering to said human or animal a composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising:

(b) a source of naturally occurring soy isoflavones, in the form of a soy protein source with a high concentration of naturally occurring isoflavones and/or a soy germ extract, said source of naturally occurring isoflavones providing naturally occurring isoflavones in an amount of minimum 50 mg and sufficient to ensure that the total content of isoflavones of the composition amounts to at least 0.34 weight percent but less than 2.50 weight percent of the soy protein content of the composition, thereby reducing the influx of cholesterol and/or triglycerides into the arterial wall and/or causing dilation of blood vessels and/or reducing restriction of blood vessels and/or reducing the amount of oxidized LDL-cholesterol present in the arterial wall and/or preventing, reducing or eliminating fatty streak formation and/or preventing, reducing or eliminating fibrous plaque formation.

14. A method of improving by therapy any of the following in a human or animal subject: physical performance and/or athletic performance by a human or animal by at least 5% as measured in watts in bicycling tests for 45 min at 130 pulse rate per minute, or vascularization and conditioning of the skin measured as diminishing and/or smoothening of wrinkles, said method comprising administering to said human or animal a composition having a total protein content providing at least 15 percent of the total energy content of the composition, and comprising: (a) a soy protein source, which provides for at least 12.5 g of soy protein and at least 50 weight percent of the total protein content of the composition,

15. A method according to claim 13 or 14, wherein said composition further comprises:

16. A method according to any of claims 13-15, wherein said method results in the administration of at least 100 mg of said naturally occurring isoflavones to said human or animal subject on a daily basis.

17. A method according to any of claims 13, 15 or 16 wherein the micro cardiovascular disease is an arteriosclerotic condition of the human or animal body.

18. A method according to claim 17 wherein the micro cardiovascular disease is selected from the group comprising arteriosclerosis, atherosclerosis, arteriolosclerosis and capillary sclerosis.