RU2820163C2 - Compositions and methods using at least one glycine or derivative thereof, at least one n-acetylcysteine or derivative thereof and at least one nicotinamide riboside or nad+ precursor - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to methods and uses of compositions for treating, reducing the incidence or severity of metabolic syndrome, age-related decrease in metabolic regulation and muscle atrophy, for delaying the onset of harmful effects of aging and osteoporosis in healthy older adults. Composition contains an effective amount of a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside. Composition can be administered orally, for example, in the form of one or more of a food product, a food product for special medical purposes (FSMP), a food additive, a ready-to-eat mixture, a milk beverage, liquid additive of small volume, powder forms for dilution with liquid or food substitute beverage.

EFFECT: invention allows treating or preventing oxidative stress and conditions associated with oxidative stress.

21 cl, 1 dwg, 2 ex

Description

Prerequisites for creating the invention

The present description essentially relates to compositions and methods capable of treating or preventing oxidative stress, a condition associated with oxidative stress, reduced glutathione levels, or a condition associated with reduced glutathione levels. Alternatively or additionally, the compositions and methods may improve exercise capacity and physical function.

Population aging is a noticeable demographic phenomenon. As the growth of the elderly population has exceeded the overall population growth due to increasing life expectancy, the proportion of elderly people relative to the rest of the population has increased significantly. For example, in 1950, one in twelve people was at least 60 years old, and by the end of 2000, one in ten were aged 60 or older. By the end of 2050, one in five people in the world is projected to be 60 years of age or older.

Elderly or aging individuals often suffer from some degree of cognitive impairment, including cognitive decline that progresses with age, and age-related changes in brain morphology and cerebrovascular function are common. With age, there is a consistent decline in cognitive abilities across a range of cognitive functions, including processing speed, attention, episodic memory, spatial awareness, and goal-directed activity. Brain imaging studies have shown that these normal age-related cognitive declines are associated with a decrease in the volume of both gray and white matter in the brain, with the frontostriatal system most often affected by aging. This decrease in cortical volume can be explained by a number of destructive cellular processes associated with normal aging, such as the accumulation of free radical damage over time leading to oxidative damage, chronic low-grade inflammation, accumulation of homocysteine (increased levels of which are a risk factor for cognitive decline and dementia) and decreased mitochondrial efficiency. Along with direct cellular damage, the brain is also indirectly damaged due to effects on microvascular structures. It is obvious that the pathology of aging, as well as dementia, is a complex of these interacting factors related to each other. For example, mitochondrial dysfunction leads to increased oxidative stress, and oxidative stress can cause inflammation and vascular damage.

Additionally, cognitive decline is an early prognostic factor or pathology of Alzheimer's disease, and it begins before the onset of dementia. In this context, a comprehensive cognitive assessment provides a reliable means of assessing cognitive decline that precedes dementia. Reliable evidence suggests that maintaining brain health and preventing cognitive decline with age can prevent or slow the progression of dementia caused by Alzheimer's disease and other age-related neuropathologies.

In biology and psychology, the term "stress" refers to the consequences of a human or other animal's failure to respond appropriately to physiological, emotional, or physical threats, whether actual or imagined. The psychobiological features of stress can manifest as signs of oxidative stress, that is, an imbalance between the production and expression of reactive oxygen species and the ability of the biological system to quickly eliminate the toxic effects of reactive intermediates or repair the resulting damage. Disturbances in the normal redox status of tissues can lead to toxic effects through the production of peroxides and free radicals that damage all cellular components, including proteins, lipids, and DNA. Some active oxidizing agents can even act as messengers through a phenomenon called “redox signaling.”

In humans, oxidative stress is involved in many diseases. Examples include atherosclerosis, Parkinson's disease, heart failure, myocardial infarction, Alzheimer's disease, schizophrenia, bipolar disorder, fragile X syndrome, and chronic fatigue syndrome.

One source of active oxygen under normal conditions in humans is the leakage of activated oxygen from mitochondria through oxidative phosphorylation. Other enzymes capable of producing superoxide (O2-) are xanthine oxidase, NADPH oxidases, and cytochrome P450s. Hydrogen peroxide, another strong oxidizing agent, is generated by a wide range of enzymes, including several oxidases. Reactive oxygen species play an important role in cellular signaling called redox signaling. Thus, to maintain proper cellular homeostasis, it is necessary to maintain a balance between the production and consumption of active oxygen.

Oxidative stress contributes to tissue damage after irradiation and hyperoxia. It is thought to play an important role in neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and Huntington's disease.

Oxidative stress is also thought to be associated with certain cardiovascular diseases because oxidation of low-density lipoprotein (LDL) in the vascular endothelium is a precursor to plaque formation. Oxidative stress also plays a role in the ischemic cascade due to oxygen reperfusion injury following hypoxia. This cascade includes both strokes and heart attacks. Oxidative stress also contributes to chronic fatigue syndrome.

Moreover, the free radical theory of aging suggests that the biological process of aging leads to increased oxidative stress in older adults. The ability of cells to resist the damaging potential of oxidative stress is determined by the vital balance between the production of oxidative free radicals and the range of protective antioxidants available to the cell. There are many antioxidant defense systems, and glutathione (GSH) is the most abundant intracellular component of all antioxidant defenses. GSH, a tripeptide, is synthesized from the precursor amino acids glutamate, cysteine, and glycine in two steps catalyzed by glutamate cysteine ligase (GCL, also known as gamma-glutamylcysteine synthetase, EC 6.3.2.2) and gamma-L-glutamyl-L-cysteine: glycyligase (also known as glutathione synthetase, EC 6.3.2.3), and GSH synthesis occurs de novo in cells.

Nicotinamide riboside can stimulate increased intracellular levels of nicotinamide adenine dinucleotide (NAD ⁺ ) in cells and tissues to improve cell and tissue survival. Moreover, the combination of glycine and N-acetylcysteine (NAC) can be used for therapeutic and/or prophylactic indications in mammals in need. For example, a combination of glycine and N-acetylcysteine (NAC) may provide beneficial effects in the event of a disorder of glutathione metabolism, and/or an increase in oxidative stress, and/or oxidative damage in a mammal, including such a disorder of glutathione metabolism, and/or an increase in oxidative stress, and /or oxidative damage due to aging or diabetes, and can be combined with nicotinamide riboside.

Additionally, with regard to NAD ⁺ , which can be increased within cells using the findings herein (eg, by combining glycine and NAC), all living cells use NAD pathways, and NAD ⁺ is the oxidized form of NAD. In metabolism, NAD is involved in redox reactions and transfers electrons from one reaction to another. NAD ⁺ accepts electrons from other molecules and is reduced to form NADN, which can then be used as a reducing agent to donate electrons. These electron transfer reactions are the main function of NAD, but it is also used in other cellular processes, particularly as a substrate for enzymes that add or remove chemical groups from proteins during post-translational modifications.

Summary of the invention

Various combinations of compounds described herein reduce oxidative stress and improve mitochondrial function (both of these processes are related in their mechanisms). Mitochondrial dysfunction contributes to cellular damage, in part through reactive oxygen species (ROS) and metabolic dysfunction (inability to metabolize nutrients), which in turn leads to metabolic and degenerative diseases. Without being limited by any theory, the present inventors believe that various combinations of the compounds described herein, by reducing oxidative stress and improving mitochondrial function, can treat, reduce the incidence of, or reduce the severity of such metabolic and degenerative diseases, at least having a combined effect, and possibly reinforcing each other or providing synergy.

Accordingly, in a general embodiment, provided herein is a method of treating or preventing at least one physical condition selected from the group consisting of oxidative stress, a condition associated with oxidative stress, a reduced glutathione level, and a condition associated with a reduced glutathione level. The method includes administering to an individual in need thereof an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In one embodiment, the method includes administering to an individual in need thereof an effective amount of a combination of at least one glycine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor.

In one embodiment, the method includes administering to an individual in need thereof an effective amount of a combination of at least one N-acetylcysteine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor.

In one embodiment, the at least one physical condition is selected from the group consisting of the deleterious effects of aging, muscle wasting, prediabetes, gestational diabetes, type I diabetes, type II diabetes, complications of diabetes, insulin resistance, metabolic syndrome, dyslipidemia, overweight , obesity, elevated cholesterol levels, elevated triglyceride levels, elevated fatty acid levels, fatty liver, kidney disease, cardiovascular disease, neurodegenerative disease, cognitive impairment, myopathy such as statin-induced myopathy, non-alcoholic steatohepatitis, tinnitus, dizziness, hangover, hearing impairment, osteoporosis, hypertension, atherosclerosis/coronary artery disease, myocardial damage after stress, traumatic brain injury, cystic fibrosis, inflammation, cancer and HIV infection.

In one embodiment, at least one glycine or a functional derivative thereof is selected from the group consisting of L-glycine, L-glycine ethyl ester, D-allylglycine; N-[bis(methylthio)methylene]glycine methyl ester; Boc-allyl-Gly-OH (dicyclohexylammonium) salts; Boc-D-Chg-OH; Boc-Chg-OH; (R)-N-Boc-(2'-chlorophenyl)glycine; Boc-L-cyclopropylglycine; Boc-L-cyclopropylglycine; (R)-N-Boc-4-fluorophenylglycine; Boc-D-propargylglycine; Boc-(S)-3-thienylglycine; Boc-(R)-3-thienylglycine; D-a-cyclohexylglycine; L-a-cyclopropylglycine; N-(2-fluorophenyl)-N-(methylsulfonyl)glycine; N-(4-fluorophenyl)-N-(methylsulfonyl)glycine; Fmoc-N-(2,4-dimethoxybenzyl)-Gly-OH; N-(2-furoyl)glycine; L-a-neopentylglycine; D-propargylglycine; sarcosine; Z-a-phosphonoglycine trimethyl ester and mixtures thereof.

In one embodiment, the combination is administered orally.

In one embodiment, the combination is administered in a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-use formula, a milk drink, a small volume liquid supplement, powder forms for reconstitution with liquid, meal replacement drink and combinations thereof.

In an embodiment, the combination is administered in a composition that contains a dipeptide that provides at least a portion of at least one glycine or a functional derivative thereof and at least one N-acetylcysteine or a functional derivative thereof.

In one embodiment, at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor are administered in the same composition.

In one embodiment, one or more of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor is administered in a different composition from the rest of the combination.

In another embodiment, provided herein is a method for slowing the onset of metabolic abnormalities, maintaining muscle mass, reducing oxidative stress, maintaining immune function, and/or maintaining cognitive function in a healthy older adult. The method includes administering to a healthy older adult an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. A healthy older adult may be elderly.

In another embodiment, provided herein is a method of enhancing reactive oxygen species metabolism, improving glucose control, and/or improving muscle function in an individual with at least one of obesity, prediabetes, or diabetes. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In another embodiment, provided herein is a method of improving mitochondrial function in an individual suffering from sarcopenia. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. An individual with sarcopenia may be otherwise healthy.

In another embodiment, provided herein is a method of improving one or more of fetal metabolic programming to prevent the subsequent development of obesity, prediabetes and/or diabetes, maternal and fetal health in gestational diabetes, exercise capacity and physical function, quality of life, life expectancy , memory, cognitive function, post-traumatic recovery and survival or recovery from injury and surgery. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. The individual may have at least one of cognitive impairment or cognitive impairment, and the composition may contain an amount of the combination effective to improve cognitive function. The individual may not have cognitive impairment or cognitive impairment, and the composition may contain an amount of the combination effective to improve cognitive function.

In another embodiment, provided herein is a method for improving at least one of muscle performance or muscle recovery after exercise. The method includes administering to an exercise-trained individual in need thereof an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor in at least one of the time periods selected from the group consisting of time before physical activity, time during physical activity and time after physical activity.

In another embodiment, provided herein is a composition comprising a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. The composition may also contain a combination of at least one glycine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor. The composition may also contain a combination of at least one N-acetylcysteine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor.

The composition contains a combination in an amount effective for at least one of (i) treating or preventing at least one physical condition selected from the group consisting of oxidative stress, a condition associated with oxidative stress, decreased glutathione levels, a condition associated with reduced glutathione levels, or (ii) improving one or more of fetal metabolic programming to prevent the subsequent development of obesity, prediabetes and/or diabetes, maternal and fetal health in gestational diabetes, exercise capacity and physical function, quality of life, life expectancy, memory, cognitive function, post-traumatic recovery and survival or recovery from trauma and surgery.

In one embodiment, the amount of the combination is effective for treating or preventing at least one physical condition selected from the group consisting of the harmful effects of aging, muscle wasting, prediabetes, gestational diabetes, type I diabetes, type II diabetes, complications of diabetes, resistance to insulin, metabolic syndrome, dyslipidemia, overweight, obesity, elevated cholesterol levels, elevated triglyceride levels, elevated fatty acid levels, fatty liver, kidney disease, cardiovascular disease, neurodegenerative disease, cognitive impairment, myopathy such as that caused by statins myopathy, non-alcoholic steatohepatitis, tinnitus, dizziness, hangover syndrome, hearing impairment, osteoporosis, hypertension, atherosclerosis/coronary artery disease, myocardial damage after stress, traumatic brain injury, cystic fibrosis, inflammation, cancer and HIV infection.

In one embodiment, the composition is selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-drink formula, a milk beverage, a small volume liquid supplement, powdered forms for liquid reconstitution, a meal replacement beverage, and their combinations.

In another embodiment, provided herein is a method of achieving at least one result selected from the group consisting of (i) reducing the severity and/or incidence of the effects of aging, (ii) maintaining or improving cellular function and/or general health, (iii) maintaining at least one of normal mitochondrial function, cell protection or energy metabolism, (iv) increasing daily energy levels, (v) reducing fatigue, (vi) maintaining or improving physical energy and/or cognitive performance, (vii) promoting healthy aging by promoting healthy or normal cellular function, (viii) maintaining healthy skin, (ix) treating heart failure and/or reducing the severity or incidence of heart failure, (x) treating, reducing the incidence or severity of oxidative stress and/ or decreased glutathione (GSH) levels occurring during a period including an intensive care unit (ICU) stay, (xi) treatment, decreased incidence, or decreased severity of another condition associated with oxidative stress and/or decreased GSH levels, ( xii) promoting recovery from injury, illness or surgery, (xiii) modulating NAD ⁺ levels in a patient with cancer or cancer in remission, (xiv) treating, reducing the incidence or reducing the severity of bariatric surgery symptoms, (xv) treating, reducing the incidence or reduction of the severity of non-alcoholic fatty liver disease (NAFLD), (xvi) treating, reducing the incidence or reduction of the severity of infection caused by the human immunodeficiency virus (HIV), and (xvii) combinations thereof, the method comprising administering to the individual an effective amount of the combination at at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In another embodiment, provided herein is a composition comprising a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor, wherein the composition contains the combination in an amount effective for at least one of (i) reducing the severity and/or incidence of the effects of aging, (ii) maintaining or improving cellular function and/or general health, (iii) maintaining at least one of normal mitochondrial function, protection cells or energy metabolism, (iv) increasing daily energy levels, (v) reducing fatigue, (vi) maintaining or improving physical energy and/or cognitive performance, (vii) promoting healthy aging by promoting healthy or normal cellular function, (viii) maintaining skin health, (ix) treating heart failure and/or reducing the severity or incidence of heart failure, (x) treating, reducing the incidence or reducing the severity of oxidative stress and/or reducing glutathione (GSH) levels occurring during a period including stay in an intensive care unit (ICU), (xi) treating, reducing the incidence or severity of another condition associated with oxidative stress and/or reduced GSH, (xii) promoting recovery from injury, illness or surgery, (xiii) ) modulating NAD ⁺ levels in a patient with cancer or cancer in remission, (xiv) treating, reducing the incidence or reducing the severity of symptoms of bariatric surgery, (xv) treating, reducing the incidence or reducing the severity of non-alcoholic fatty liver disease (NAFLD), ( xvi) treating, reducing the incidence or reducing the severity of infection caused by the human immunodeficiency virus (HIV).

One or more embodiments presented herein have the advantage of promoting beneficial effects on oxidative metabolism and preventing DNA damage.

Another benefit of one or more embodiments presented herein is the restoration of NAD ⁺ pools that are depleted with age.

Another benefit of one or more embodiments presented herein is to help slow the metabolic changes associated with aging.

Another benefit of one or more embodiments presented herein is to promote increased fatty acid metabolism.

One or more embodiments presented herein have the benefit of helping to maintain heart health, treat heart failure, and/or reduce the severity or incidence of heart failure.

Another benefit of one or more embodiments presented herein is to help maintain healthy levels of LDL cholesterol and fatty acids in the blood.

Another benefit of one or more embodiments presented herein is the replenishment of essential amino acids, sufficient quantities of which become less available to cells during the aging process.

One or more embodiments presented herein have the advantage of providing amino acids that are precursors to glutathione production, which is important for cell function.

Another benefit of one or more embodiments presented herein is to help increase glutathione levels in cells.

Another advantage of one or more embodiments proposed in the present invention is to increase the concentration of glutathione, which decreases with age.

One or more embodiments presented herein have the advantage of helping to maintain healthy muscle mass.

Another benefit of one or more embodiments presented herein is to help reduce oxidative stress in the body.

Another benefit of one or more embodiments provided herein is the support of the normal immune system by modulating glutathione levels.

Additional features and benefits are described herein and will be apparent upon viewing the following figures and reading the detailed description.

Brief description of graphic materials

Fig. 1 is a graph of data from an experimental example described herein.

Fig. 1. Effect of a combination of NAC, glycine and nicotinamide riboside (NR) on mitochondrial respiration. High-resolution respirometry of mitochondria isolated from adult zebrafish muscle (N = 5–6/condition). Acute oxidative stress was induced by treatment with menadione solution (Men) for 2 hours. Respiratory recovery was assessed by treatment for 2 h with low dose NAC + glycine (glyNAC; NAC 100 μM, glycine 100 μM) and low dose NR (50 μM). Data from respiration measurements are shown in 1 A. The theoretical additive effect (ΔglyNAC + ΔNR) was calculated as the sum of the average of glyNAC and NR and compared with the synergistic effect (ΔglyNAC + NR) (1B). Men = control with menadione; NAC = N-acetyl-cysteine; gly = glycine, NR = nicotinamide riboside. * p < 0.05, ** p < 0.01, **** p < 0.0001

Detailed description

Definitions

Below are some definitions. However, definitions may appear below in the “Embodiments” section, and the above heading “Definitions” does not mean that the above descriptions in the “Embodiments” section are not definitions.

Unless otherwise indicated, all percentages herein are expressed by weight of the total weight of the composition. As used herein, the terms "about", "approximately" and "substantially" refer to numbers in a numerical range, such as -10% to +10% of the number, preferably -5% to +5% of said number, more preferably -1% to +1% of said number, and most preferably -0.1% to +0.1% of said number. It should be understood that all numeric ranges herein include all integers and fractions included in the range. Moreover, these numerical ranges should be considered to support the claim referring to any number or subset of numbers within that range. For example, the description "from 1 to 10" should be considered to include the range "from 1 to 8", "from 3 to 7", "from 1 to 9", "from 3.6 to 4.6", "from 3, 5 to 9.9", etc.

As used in this specification and the accompanying claims, the singular forms include plural references unless the context clearly requires otherwise. Thus, for example, the reference to “one component” or “this component” implies the presence of two or more components.

The words “contain,” “contains,” and “comprising” should be interpreted as inclusive and not exclusive. Likewise, the terms “include,” “including,” and “or” should be interpreted in an inclusive sense unless such interpretation is clearly inconsistent with the context. However, the compositions described herein may not contain any element not specifically described herein. Thus, a description of an embodiment using the term “comprising” includes a description of embodiments “consisting essentially of” and “consisting of” the specified components. A composition "consisting essentially of" contains at least 50% by weight of said components, preferably at least 75% by weight of said components, more preferably at least 85% by weight of said components, most preferably at least 95 wt.% of these components.

The term "and/or" when used in the context of "X and/or Y" should be interpreted as "X" or "Y" or "X and Y". Likewise, “at least one of X or Y” should be interpreted as “X” or “Y” or “X and Y”. For example, “at least one glycine or a functional derivative thereof” should be interpreted as “glycine” or “a functional derivative of glycine” or “both glycine and a functional derivative of glycine.”

As used herein, the terms “for example” and “such as,” particularly when followed by a listing of terms, are used for exemplary and illustrative purposes only and should not be considered exclusive or exhaustive. As used herein, a condition “associated with” or “caused by” another condition means that the conditions occur simultaneously, preferably means that the conditions are caused by the same underlying condition, and most preferably means that one of the specified conditions is caused by another the specified condition.

The terms "food product", "food product" and "food composition" mean a food product or composition that is intended for ingestion by an individual, such as a human, and provides that individual with at least one nutrient. The food product typically includes at least one of a protein, a lipid, a carbohydrate, and optionally includes one or more of vitamins and minerals. The compositions of the present invention, including many embodiments described herein, may contain, consist of or consist essentially of the elements described herein, as well as any additional or optional ingredients, components or elements described herein or otherwise manner suitable for use in the diet.

As used herein, the term “isolated” means separated from one or more other compounds or components with which the compound may otherwise be found, such as found in nature. For example, “isolated” preferably means that the compound is separated from at least a portion of the cellular material with which it is typically found in nature. In one embodiment, the isolated compound is pure, i.e. does not contain any other compound.

“Prevention” includes reducing the risk and/or severity of a condition or disorder. The terms “treat,” “cure,” and “alleviate” include both prophylactic or preemptive treatment (which prevents and/or slows the progression of the target condition or disorder) and curative, therapeutic, or disease-modifying treatment, including therapeutic measures that cure, slow, reduce symptoms and/or stop the progression of a diagnosed pathological condition or disorder; and treating patients at risk of contracting a disease or suspected of contracting a disease, as well as patients who have or have been identified as suffering from a disease or medical condition. The term does not necessarily imply that the subject receives treatment until complete recovery. The terms “treating” and “treating” also refer to maintaining and/or promoting health in an individual who does not suffer from a disease, but who may be susceptible to developing a disease condition. The terms “treating,” “treating,” and “alleviating” should also include potentiation or other enhancement of one or more primary preventive or therapeutic measures. The terms “treating,” “treating,” and “alleviating” are further intended to include treating a disease or condition through diet and diet, or preventing or preventing a disease or condition through diet and diet. Treatment can be viewed from the patient's or physician's perspective.

As used herein, the term “unit dosage form” refers to physically discrete units suitable for use as unitary dosages for human and animal subjects, each unit containing a predetermined amount of a composition described herein in an amount sufficient to achieve the desired effect , in combination with a pharmaceutically acceptable diluent, carrier or carrier medium. Specifications for a unit dosage form depend on the specific compounds used, the effect achieved, and the pharmacodynamics of each compound in the host.

The "subject" or "individual" is a mammal, preferably a human. The term “elderly” in the context of a person means at least 60 years of age from birth, preferably over 63 years of age, more preferably over 65 years of age, and most preferably over 70 years of age. The term "older adult" in a human context means at least 45 years of age from birth, preferably greater than 50 years, more preferably greater than 55 years, and includes older individuals.

As used herein, the term “effective amount” means an amount that effectively prevents a deficiency, treats a disease or medical condition in an individual, or more generally reduces symptoms, controls the progression of a disease, or provides a nutritional, physiological, or medical benefit to an individual. Relative terms “improved”, “increased”, “strengthened”, etc. refer to the effects of a composition described herein, namely a composition containing at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor, compared to a composition lacking at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor, but otherwise identical. As used herein, the term “stimulation” refers to an increase or induction relative to the level prior to administration of the composition described herein.

The term sarcopenia is defined as age-related loss of muscle mass and muscle functionality (including muscle strength and walking speed). As used herein, frailty is defined as a clinically recognizable state of increased vulnerability resulting from aging-related decline in the reserves and functions of multiple physiological systems, resulting in an impaired ability to cope with daily or acute stressors. In the absence of an established quantitative standard, frailty is functionally defined by Fried et al. as meeting three of five phenotypic criteria indicating energy impairment: (1) weakness (grip strength initially in the bottom 20% of the population, adjusted for sex and body mass index), (2) poor endurance and energy reserve (self-reported wasting associated with V̇O ₂ max), (3) slowness (baseline in the bottom 20% of the population based on time required to walk 15 feet adjusted for sex and standing height), (4) low physical activity (weighted estimate of kilocalories burned per week at baseline, lowest physical activity quintile identified for each sex; for example, less than 383 kcal/week for men and less than 270 kcal/week for women), and/or unintentional weight loss (10 pounds in the past year) . Fried LP, Tangen CM, Walston J, et al., “Frailty in older adults: evidence for a phenotype.” J. Gerontol. A. Biol. Sci. Med. Sci. 56(3):M146–M156 (2001). A pre-frailty stage in which one or two of these criteria is present identifies a high risk of progression to frailty.

“Cachexia” is a severe condition of body wasting characterized by severe weight loss, anorexia, asthenia and anemia. Cachexia is a common feature of a number of diseases, such as cancer, sepsis, chronic heart failure, rheumatoid arthritis, and acquired immunodeficiency syndrome (AIDS).

A person is defined as “overweight” if their body mass index (BMI) is between 25 and 30 kg/ ^m2 . "Obesity" in a person is defined as a BMI of at least 30 kg/m ² , for example 30-39.9 kg/m ² . “Weight loss” is a decrease in total body weight. Weight loss may, for example, refer to the loss of total body weight for the purpose of improving one or more of health, fitness or appearance.

The term diabetes covers both type I and type II forms of the disease. Non-limiting examples of risk factors for diabetes include: waist circumference greater than 40 inches in men or 35 inches in women, blood pressure 130/85 mm Hg. Art. or higher, triglycerides more than 150 mg/dL, fasting blood glucose more than 100 mg/dL, or high-density lipoprotein less than 40 mg/dL in men or 50 mg/dL in women.

As used herein, the term “metabolic syndrome” refers to a combination of medical disorders that, when co-occurring, increase the risk of developing cardiovascular disease and diabetes. It affects one in five people in the United States, and the prevalence increases with age. Some studies have found the prevalence in the United States to be 25% of the population. According to the international consensus definition of the International Diabetes Foundation (2006), metabolic syndrome is the central type of obesity plus any two of the following factors.

Elevated triglycerides: > 150 mg/dL (1.7 mmol/L) or specific treatment for such lipid abnormality;

reduced HDL cholesterol: <40 mg/dL (1.03 mmol/L) in men, <50 mg/dL (1.29 mmol/L) in women, or specific treatment for this lipid abnormality;

high blood pressure: systolic blood pressure > 130 or diastolic blood pressure > 85 mm Hg. Art. or treatment of previously diagnosed hypertension; And

elevated fasting plasma glucose (FPG) > 100 mg/dL (5.6 mmol/L) or previously diagnosed type 2 diabetes.

As used herein, the term “neurodegenerative disease” or “neurodegenerative disorder” refers to any condition associated with the progressive loss of functional neurons in the central nervous system. In one embodiment, the neurodegenerative disease is associated with age-related cell death. Non-limiting examples of neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (also known as ALS and Lou Gehrig's disease), AIDS dementia complex, adrenoleukodystrophy, Alexander disease, Alper disease, ataxia telangiectasia, Batten disease , bovine spongiform encephalopathy (BSE), Canavan disease, corticobasal degeneration, Creutzfeldt-Jakob disease, dementia with Lewy bodies, fatal familial insomnia, frontotemporal lobar degeneration, Kennedy disease, Krabbe disease, Lyme disease, Machado Joseph disease, multiple sclerosis, multiple system atrophy, neuroacanthocytosis, Niemann-Pick disease, Pick's disease, primary lateral sclerosis, progressive supranuclear gaze palsy, Refsum's disease, Sandhoff's disease, diffuse myelinoclastic sclerosis, spinocerebellar ataxia, subacute combined degeneration of the spinal cord, tabes dorsalis, disease Tay-Sachs, toxic encephalopathy, transmissible spongiform encephalopathy and staggering hedgehog syndrome.

As used herein, the term “cognitive performance” refers to how well a subject performs one or more cognitive functions. As used herein, the term "cognitive function" refers to the mental process by which an individual learns, perceives, or makes sense of ideas. It includes all aspects of perception, thinking, reasoning and remembering and includes, for example, perception, memory, attention, speech comprehension, speech generation, reading comprehension, imagery, learning and logic. Typically it will relate to at least memory.

Methods for measuring cognitive function are well known and may include, for example, single tests or panels of tests for any aspect of cognitive function. One such test is the Prado Test of Cognitive Function, developed by Margallo-Lana et al. (2003) J. Intelligence. Disability Res. 47:488-492. Another such test is the Mini-Mental State Examination (MMSE), which is designed to assess orientation in time and space, writing, attention and computation, memory, language use and comprehension, repetition and execution of complex commands. Folstein et al. (1975) J. Psych. Res. 12:189-198. Other tests that can be used to measure cognitive function include the Alzheimer's Disease Assessment Scale (ADAS-Cog) (Rosen et al. (1984) Am. J. Psychiatry. 141(11): 1356-64) and the Cambridge Automated Neuropsychological Battery tests (CANTAB) (Robbins et al. (1994) Dementia. 5(5):266-81). Such tests can be used to objectively assess cognitive function so that changes in cognitive function can be measured and compared, for example, in response to treatment in accordance with the methods described herein.

As used herein, the term “cognitive disorder” refers to any condition that impairs cognitive function. Non-limiting examples of cognitive impairment include delirium, dementia, learning disorder, attention deficit disorder (ADD), and attention deficit hyperactivity disorder (ADHD).

Embodiments

Provided herein are compositions containing a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. As used herein, the term “nicotinamide riboside” includes L-valine and L-phenylalanine esters of nicotinamide riboside. Non-limiting examples of suitable NAD ⁺ precursors include tryptophan, nicotinic acid, nicotinamide, reduced form of nicotinamide riboside (NRH), nicotinamide mononucleotide (NMN), trinelline, nicotinic acid mononucleotide, nicotinic acid riboside, and mixtures thereof. The present invention is not limited to a particular embodiment of the NAD ⁺ precursor, and the NAD ⁺ precursor can be any NAD ⁺ stimulating compound.

Each of the compounds can be administered simultaneously with the other compounds (ie, as a single dosage unit) or separated by time (ie, in separate dosage units). The present specification further provides a kit containing at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor for mixing to form one or more compositions, described herein, and/or for use in one or more of the methods described herein, for example, in separate containers as two or more liquid solutions or dried powders. In some embodiments, one or more of these compounds may be isolated compounds.

The combination of at least one glycine or a functional derivative thereof and at least one N-acetylcysteine or a functional derivative thereof can be prepared as any of the compositions described in US Pat. Nos. 8,362,080, 8,802,730 and 9,084,760, each of which is entitled "Increasing glutathione levels" for therapy" and in WO2016/191468 entitled "Benefits of Supplementation with N-Acetylcysteine and Glycine to Improve Glutathione Levels", each of which is incorporated herein by reference in its entirety.

Nicotinamide riboside may be provided in any of the compositions described in U.S. Patent Nos. 8,383,086 and 8,197,807, each entitled "Nicotinamide riboside kinase compositions and methods for using the same", and U.S. Patent No. 8,106,184, entitled "Nicotinoyl riboside compositions and methods of use ", each of which is incorporated herein by reference in its entirety.

Accordingly, an aspect of the present disclosure is a composition containing at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor in an amount effective to treat or prevent at least a condition selected from the group consisting of the harmful effects of aging, muscle wasting (from any cause, including at least sarcopenia, HIV infection, aging, cachexia, harmful effects of weight loss), prediabetes, gestational diabetes, diabetes mellitus (I type or type II), complications of diabetes (eg, diabetic dyslipidemia and/or diabetic microvascular complications such as nephropathy, retinopathy and/or neuropathy), insulin resistance, metabolic syndrome, dyslipidemia, overweight, obesity, elevated cholesterol levels, elevated triglyceride levels, elevated fatty acid levels, fatty liver (eg, non-alcoholic fatty liver, including with or without inflammation), kidney disease, cardiovascular disease (eg, heart failure and/or cardiac dysfunction, e.g. treating heart failure and/or reducing the severity or incidence of heart failure), neurodegenerative disease (eg, as a result of aging), cognitive impairment, myopathy such as statin-induced myopathy, non-alcoholic steatohepatitis, tinnitus, dizziness, hangover syndrome, impairment hearing, osteoporosis, hypertension, atherosclerosis/coronary artery disease, myocardial damage after stress (eg, caused by burns or trauma), traumatic brain injury (including concussions), cystic fibrosis, inflammation, cancer and HIV infection. In addition, it should be noted that aging may be accompanied by a decrease in NAD ⁺ and glutathione levels. Additionally, with regard to cardiovascular disease (CVD), it should be noted that increased homocysteine levels are a well-established risk factor for CVD.

Another aspect of the present disclosure is a method of treating at least one of these conditions, comprising administering to an individual a composition comprising a therapeutically effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least at least one nicotinamide riboside or NAD ⁺ precursor. Another aspect of the present disclosure is a method of preventing at least one of these conditions, comprising administering to an individual at risk of developing at least one condition a composition comprising a prophylactically effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof and at least one nicotinamide riboside or NAD ⁺ precursor.

The composition may treat or prevent sarcopenia, sarcopenic obesity or cachexia, for example, due to an underlying medical condition such as chronic disease, HIV, cancer, chronic obstructive pulmonary disease (COPD) and/or aging in otherwise healthy individuals. In this regard, aging may be accompanied by a decrease in NAD ⁺ and glutathione (GSH) levels.

The composition may treat or prevent eye disease caused, directly or indirectly, by low levels of GSH in the lens of the eye, which is known to be rich in glutathione. Non-limiting examples of such conditions include cataracts and/or glaucoma, presbyopia (age-related near vision loss requiring glasses) and presbycusis (age-related hearing loss requiring hearing aids).

In one embodiment, the composition improves at least one of muscle performance or muscle recovery, for example, after muscle stress, including exercise-related muscle stress. Physical activity can be of any type, including, for example, aerobic (“cardiac”) exercise and/or strength training. The composition can be administered at at least one of the time periods selected from the group consisting of time before exercise (for example, less than an hour), time during exercise, and time after exercise (for example, less than an hour after physical activity).

Another aspect of the present disclosure is a method for slowing the onset of metabolic abnormalities, maintaining muscle mass, reducing oxidative stress, maintaining immune function, and/or maintaining cognitive function in a healthy older adult. The method includes administering to a healthy older adult an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

Another aspect of the present disclosure is a method of improving mitochondrial function in an individual suffering from sarcopenia. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

Another aspect of the present disclosure is a method of enhancing the metabolism of reactive oxygen species, improving glucose control and/or improving muscle function in an individual with at least one of obesity, prediabetes or diabetes. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

Another aspect of the present disclosure is a method of improving mitochondrial function (preferably to achieve a beneficial effect on at least one of metabolism or strength) in an individual suffering from sarcopenia. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In another aspect of the present disclosure, a method for improving cognitive function is provided. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor. The cognitive function may be selected from the group consisting of perception, memory, attention, speech comprehension, speech generation, reading comprehension, imagery, learning, logic, and combinations thereof. In one embodiment, the individual does not have a cognitive disorder; in an alternative embodiment, the individual has a cognitive disorder. The individual may be elderly and/or may have cognitive decline associated with aging.

Another aspect of the present disclosure is a method of improving one or more of fetal metabolic programming to prevent the subsequent development of obesity, prediabetes and/or diabetes, maternal and fetal health in gestational diabetes, exercise capacity and physical function, quality of life, life expectancy, memory, cognitive function, post-traumatic recovery and survival (eg, after surgery, after sepsis, after blunt or penetrating trauma caused by accidental or assault), or recovery from trauma and surgery. The method includes administering to an individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In another aspect, provided herein is a method of achieving at least one result selected from the group consisting of (i) reducing the severity and/or incidence of the effects of aging, (ii) maintaining or improving cellular function and/or general health, ( iii) maintaining at least one of normal mitochondrial function, cellular protection or energy metabolism, (iv) increasing daily energy levels, (v) reducing fatigue, (vi) maintaining or improving physical energy and/or cognitive performance, (vii) providing healthy aging by promoting healthy or normal cellular function, (viii) maintaining healthy skin, (ix) treating heart failure and/or reducing the severity or incidence of heart failure, (x) treating, reducing the incidence or severity of oxidative stress and/or reduction in glutathione (GSH) levels occurring during a period including an intensive care unit (ICU) stay, (xi) treatment, reduction in the incidence or severity of another condition associated with oxidative stress and/or reduced GSH, (xii ) promoting recovery from injury, illness or surgery, for example rehabilitation specific to neurological conditions such as traumatic brain injury or stroke, (xiii) modulating NAD ⁺ levels in a patient with cancer or cancer in remission, (xiv) treatment, reducing the incidence or reducing the severity of symptoms of bariatric surgery, (xv) treating, reducing the incidence or reducing the severity of non-alcoholic fatty liver disease (NAFLD), (xvi) treating, reducing the incidence or reducing the severity of human immunodeficiency virus (HIV) infection, and (xvii) combinations thereof, the method comprising administering to the individual an effective amount of a combination of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

A particularly preferred embodiment is a method of treating, reducing the incidence of, or reducing the severity of at least one condition selected from the group consisting of metabolic syndrome, age-related metabolic decline, and muscle performance, the method comprising administering to a needy or at-risk individual an effective amount combinations of at least one glycine or a functional derivative thereof, at least one N-acetylcysteine or a functional derivative thereof, and at least one nicotinamide riboside or NAD ⁺ precursor.

In each of the compositions and methods described herein, the composition is preferably a food product, including dietary supplements, food ingredients, functional foods, dietary supplements, health foods, nutraceuticals, or dietary supplements. For example, the composition may be selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-feed formula, a milk drink, a small volume liquid supplement (e.g., about 50 ml or less, e.g., about 30 ml or less), liquid powders, meal replacement drinks, and combinations thereof.

As used herein, a "functional derivative" of glycine is a functional derivative of glycine that is effective in combination with N-acetylcysteine or a functional derivative thereof in increasing intracellular GSH levels in an individual. A “functional derivative” of N-acetylcysteine is a functional derivative of N-acetylcysteine that is effective in an individual, alone or in combination with glycine (or a functional derivative thereof) to increase intracellular GSH levels.

The glycine is preferably L-glycine and/or L-glycine ethyl ester. Non-limiting examples of suitable functional glycine derivatives include D-allylglycine; N-[bis(methylthio)methylene]glycine methyl ester; Boc-allyl-Gly-OH (dicyclohexylammonium) salt; Boc-D-Chg-OH; Boc-Chg-OH; (R)-N-Boc-(2'-chlorophenyl)glycine; Boc-L-cyclopropylglycine; Boc-L-cyclopropylglycine; (R)-N-Boc-4-fluorophenylglycine; Boc-D-propargylglycine; Boc-(S)-3-thienylglycine; Boc-(R)-3-thienylglycine; D-a-cyclohexylglycine; L-a-cyclopropylglycine; N-(2-fluorophenyl)-N-(methylsulfonyl)glycine; N-(4-fluorophenyl)-N-(methylsulfonyl)glycine; Fmoc-N-(2,4-dimethoxybenzyl)-Gly-OH; N-(2-furoyl)glycine; L-a-neopentylglycine; D-propargylglycine; sarcosine; Z-a-phosphonoglycine trimethyl ester and mixtures thereof. In an embodiment, both glycine and N-acetylcysteine may be provided in a dipeptide such as N-acetylcysteinylglycine or cysteinylglycine.

The composition can be administered at least one day per week, preferably at least two days per week, more preferably at least three or four days per week (for example, every other day), most preferably at least five days per week, six days a week or seven days a week. The administration period may be at least one week, preferably at least one month, more preferably at least two months, most preferably at least three months, such as at least four months. In one embodiment, administration is performed at least daily; for example, the subject may receive one or more doses daily. In some embodiments, administration continues for the remainder of the individual's life. In other embodiments, administration occurs until no noticeable symptoms of the medical condition remain. In certain embodiments, administration occurs until there is detectable improvement in at least one symptom, and in other cases while relief is maintained.

Glycine and N-acetylcysteine can be prepared in a specific ratio. In some embodiments, the formulation may contain these components in the following approximate ratios: 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1: 65, 1: 70, 1: 75, 1: 80, 1: 85, 1: 90, 1: 95, 1: 100, 1: 150, 1: 200, 1: 300, 1: 400, 1: 500, 1:600, 1:750, 1:1000, and 1:10,000. In specific embodiments, the formulation may contain these components in the following percentages by weight (either the same for both glycine and N-acetylcysteine, or different percentages by weight shares for each of them): for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15%, 20%, 25% , 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97% or 99%.

N-acetylcysteine or a functional derivative thereof can be administered in an amount of about 0.1-100 milligrams (mg) of N-acetylcysteine (NAC) or a functional derivative thereof per kilogram (kg) of the subject's body weight. Glycine (GLY) or a functional derivative thereof can be administered in an amount of about 0.1-100 milligrams (mg) of glycine or a functional derivative thereof per kilogram (kg) of body weight of the subject. In some embodiments, these amounts are provided at least in part by a dipeptide containing both N-acetylcysteine or a functional derivative thereof and glycine or a functional derivative thereof.

In a specific non-limiting example, daily dosages for a subject weighing 60 kg may be as follows:

NAC or its derivative: 6 to 6000 mg/day

GLY or its derivative: 6 to 6000 mg/day

Nicotinamide riboside: 0.001 to 1000 mg/day

The nicotinamide riboside or NAD ⁺ precursor may be administered in an amount of from about 0.001 mg/day to about 2000 mg/day, preferably from about 0.001 mg/day to about 1000 mg/day, more preferably from about 0.001 mg/day to about 750 mg/day , even more preferably from about 0.001 mg/day to about 500 mg/day, most preferably from about 0.001 mg/day to about 250 mg/day, for example from about 0.001 mg/day to about 100 mg/day, from about 0.001 mg /day to about 75 mg/day, from about 0.001 mg/day to about 50 mg/day, from about 0.001 mg/day to about 25 mg/day, from about 0.001 mg/day to about 10 mg/day or from about 0.001 mg/day to about 1 mg/day. Of course, the daily dose can be administered in portions at different times of the day. However, in any particular case, the amount of compound administered will depend on factors such as the solubility of the active ingredient, the formulation used, the condition of the subject (eg, body weight) and/or the route of administration. For example, the daily dosages of nicotinamide riboside described above are not limiting and may be different in some embodiments; in particular, the compositions described herein can be used as a critical medical special purpose product (FSMP) and may contain up to about 2.0 mg of nicotinamide riboside per day.

The compositions described herein can be administered to a subject orally or parenterally. Non-limiting examples of parenteral administration include intravenous, intramuscular, intraperitoneal, subcutaneous, intraarticular, intrasynovial, intraocular, intrathecal, topical, and inhalation. Thus, non-limiting examples of composition forms include natural products, processed foods, natural juices, concentrates and extracts, injectable solutions, microcapsules, nanocapsules, liposomes, patches, inhalations, nasal sprays, nasal drops, eye drops, sublingual tablets and sustained release formulations.

The compositions described herein can employ any of a variety of formulations for therapeutic administration. More specifically, pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers or diluents, and may be formulated in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalers , gels, microspheres and aerosols. Thus, administration of the composition can be achieved by various routes, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, transdermal and intratracheal administration. The active agent may, after administration, be in the systemic circulation or may be localized through the use of regional administration, intramural administration, or the use of an implant that functions to maintain the active dose at the site of implantation.

In pharmaceutical dosage forms, the compounds can be administered as their pharmaceutically acceptable salts. They may also be used in suitable association with other pharmaceutically active compounds. The following methods and excipients are provided by way of example only and are not intended to be limiting in any way.

In the case of oral preparations, the compounds can be used alone or in combination with appropriate additives to form tablets, powders, granules or capsules, for example with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, functional cellulose derivatives, gum arabic, corn starch or gelatins; with disintegrants such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants such as talc or magnesium stearate; and, if necessary, with diluents, buffering agents, wetting agents, preservatives and flavoring agents.

The compounds can be prepared as injectable preparations by dissolving, suspending or emulsifying them in an aqueous or non-aqueous solvent such as vegetable or other similar oils, synthetic aliphatic acid glycerides, higher aliphatic acid esters or propylene glycol; and, if necessary, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives.

The compounds can be used in an aerosol formulation for administration by inhalation. For example, the compounds can be prepared in pressurized propellants such as dichlorodifluoromethane, propane, nitrogen, and the like.

In addition, the compounds can be prepared as suppositories by mixing with various bases, such as emulsifying bases or water-soluble bases. The compounds can be administered rectally using a suppository. The suppository may include a carrier such as cocoa butter, carbowaxes and polyethylene glycols, which melt at body temperature but solidify at room temperature.

Unit dosage forms for oral or rectal administration may be provided, such as syrups, elixirs and suspensions, each dosage unit, for example a teaspoon, tablespoon, tablet or suppository, containing a predetermined amount of the composition. Likewise, unit dosage forms for injection or intravenous administration may contain the compounds in the composition as a solution in sterile water, normal saline, or other pharmaceutically acceptable carrier, each dosage unit, such as ml or L, containing a predetermined amount of the composition containing one or more connections.

Example

The following non-limiting example of possible use discusses experimental data that have been and/or will be studied to further validate the compounds, compositions and methods described herein.

Chronic oxidative stress induces changes in cellular respiration rates

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor) _ Seahorse measurement (respiration rate)

Synergistic reduction of oxidative stress and oxidative stress-related effects via glycine, NAC and NR

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Cell growth (protein concentration)

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ GSH production

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Metabolic profile

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Seahorse measurement (respiration rate)

Menadione-induced oxidative stress in HepG2 cells treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ NAPD-NAD oxidation-reduction status

Menadione-induced oxidative stress in zebrafish treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Measurement of oxidative stress

Menadione-induced oxidative stress in zebrafish treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ GSH products

Menadione-induced oxidative stress in zebrafish treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Metabolic profile

Menadione-induced oxidative stress in zebrafish treated with BSO (GSH synthesis inhibitor), NAC +/- glycine +/- NR _ Mitochondrial function (Oroboros respiration rate measurement, complex activity, etc.).

Example. Low dose combination of GlyNAC-NR induces restoration of mitochondrial respiration after acute oxidative stress.

Materials and methods

Danio rerio

Adult AB zebrafish were grown at 28°C under standard rearing conditions. Fish aged 4 months in water were exposed to acute oxidative stress by incubation with 3 μM menadione (Sigma-Aldrich) for 2 h at 28°C. Oxidative stress controls were then incubated in pure water for 2 h at 28°C. Fish were treated by incubating in water containing the compound of interest for 2 h at 28°C. The following compounds were used: N-acetylcysteine (NAC) (Sigma-Aldrich); glycine (Sigma-Aldrich); nicotinamide riboside (NR) (Chromadex).

Mitochondrial isolation and high-resolution respirometry

Crude mitochondrial extracts were prepared from fish trunk muscles according to a previously described method with minor modifications (Frezza et al., 2007). After quantification of BCA protein concentration, 150 μg of crude extract was used for quantification by high-resolution respirometry. An Oxygraph-2k device (O2k, OROBOROS Instruments) was used for respiration measurements. Up to two O2k devices (four chambers) were used in parallel. Experiments were performed at 28°C in modified MiR05 medium (110 mM sucrose, 0.5 mM EGTA, 3 mM _MgCl2 , 20 mM taurine, 10 mM _{KH2PO4 ,} 20 mM HEPES and 0.1% BSA, essentially free of fatty acids). Respiration of isolated mitochondria was determined using substrate-uncoupler-inhibitor titration (SUIT) protocols (Pesta and Gnaiger, 2012) with modifications. Pyruvate, glutamate, and malate (5 mM, 10 mM, 2 mM, respectively) were used as substrates to induce respiration in complex I (CI) in the presence of ADP (1 mM). The addition of succinate (10 mM) in the presence of ADP was used to induce respiration in complex II (CII). CI respiration was calculated as the difference between total respiration (CI + CII) and respiration with the addition of the CI inhibitor rotenone (0.5 μM); CII respiration was calculated as the difference between respiration with CI inhibition and respiration with the addition of the CII inhibitor malonic acid (5 mM). Total respiration (CI + CII, total respiration) was defined as the difference between respiration in the presence of all substrates and complete inhibition of CI and CII.

Statistics

All numerical data are expressed as mean ± std. osh. average and are shown as histograms. For statistical analysis, 2-factor analysis of variance (ANOVA) with Tukey's post hoc test was used, and differences were considered statistically significant at p < 0.05.

results

In fig. Figure 1 shows the effect of the combination of GlyNAC and NR on high-resolution respirometry. Fish (N = 5–6/group) were treated for 2 h with low doses of NAC (100 μM), glycine (100 μM), and NR (50 μM), and the individual effects of GlyNAC and NR were compared with a combination of the three compounds. The selected concentrations do not have any significant effect when used alone. Thus, in FIG. 1A shows that the low-dose combination significantly improved CI respiration and total respiration compared with menadione control and the compounds alone. In fig. 1B shows the synergistic effect calculated as the difference between the values for each group and the mean value of the menadione control. The theoretical additive effect (ΔGlyNAC + ΔNR) was calculated as the sum of the means for the ΔGlyNAC and ΔNR conditions. The effect of the combined effect (ΔGlyNAC + NR) is significantly higher compared to the individual compounds (ΔGlyNAC and ΔNR), and there is a trend compared to the additive effect (ΔGlyNAC + ΔNR), but not significant.

It should be understood that various changes and modifications to the currently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present subject matter and without diminishing its intended advantages. Accordingly, the appended claims are intended to cover such changes and modifications.

Claims (21)

1. A method of reducing oxidative stress and improving mitochondrial function to treat, reduce the incidence or reduce the severity of metabolic syndrome, age-related metabolic decline and muscle wasting, the method comprising administering to a needy or at-risk individual an effective amount of a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside. 2. The method according to claim 1, wherein at least one glycine is L-glycine. 3. The method according to claim 1, wherein the combination is administered orally. 4. The method of claim 1, wherein the combination is administered in a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-use formula, a milk drink, a small volume liquid supplement, powdered forms for dilution with liquid, food replacement drinks and combinations thereof. 5. The method of claim 1, wherein the combination is administered in a composition that contains a dipeptide that provides at least a portion of at least one glycine and at least one N-acetylcysteine. 6. The method of claim 1, wherein at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside are administered in the same composition. 7. The method of claim 1, wherein one or more of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside is administered in a different composition from the rest of the combination. 8. The use of a composition for treating, reducing the incidence or reducing the severity of metabolic syndrome and age-related metabolic decline, where the composition contains a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside, wherein the composition contains a combination of an amount effective to treat, reduce the incidence of, or reduce the severity of at least one condition selected from the group consisting of metabolic syndrome, age-related metabolic decline, and muscle wasting. 9. Use according to claim 8, wherein the composition is selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-use formula, a milk drink, a small volume liquid supplement, powder forms for reconstitution with liquid , food replacement drinks and their combinations. 10. A method of reducing oxidative stress and improving mitochondrial function to treat or prevent at least one physical condition selected from the group consisting of the harmful effects of aging and osteoporosis, the method comprising administering to an individual in need thereof an effective amount of a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside. 11. The method of claim 10, wherein the at least one glycine is L-glycine. 12. The method of claim 10, wherein the combination is administered orally. 13. The method of claim 10, wherein the combination is administered in a composition selected from the group consisting of a food product, a food for special medical purposes (FSMP), a dietary supplement, a ready-to-use formula, a milk drink, a small volume liquid supplement, powdered forms for dilution with liquid, food replacement drinks and combinations thereof. 14. The method of claim 10, wherein the combination is administered in a composition that contains a dipeptide that provides at least a portion of at least one glycine and at least one N-acetylcysteine. 15. The method of claim 10, wherein at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside are administered in the same composition. 16. The method of claim 10, wherein one or more of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside is administered in a different composition from the rest of the combination. 17. A method of reducing oxidative stress and improving mitochondrial function to delay the onset of the harmful effects of aging, muscle wasting, metabolic syndrome or osteoporosis in a healthy older adult, the method comprising administering to the healthy older adult an effective amount of a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside. 18. The method of claim 17, wherein the healthy older adult is elderly. 19. The use of a composition for treating, reducing the incidence or reducing the severity of metabolic syndrome and age-related decline in metabolic regulation, where the composition contains a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside, wherein the composition contains a combination of an amount effective for at least one of treating or preventing at least one physical condition selected from the group consisting of the harmful effects of aging, muscle wasting, metabolic syndrome, osteoporosis. 20. A method of reducing oxidative stress and improving mitochondrial function to achieve at least a reduction in the severity and/or incidence of the harmful effects of aging, the method comprising administering to an individual an effective amount of a combination of at least one glycine, at least one N-acetylcysteine, and at least at least one nicotinamide riboside. 21. The use of a composition for treating, reducing the incidence or reducing the severity of metabolic syndrome and age-related decline in metabolic regulation, where the composition contains a combination of at least one glycine, at least one N-acetylcysteine and at least one nicotinamide riboside, wherein the composition contains a combination of an amount effective to at least reduce the severity and/or incidence of the harmful effects of aging.