Composition and method for enhancing the effects of one or more anabolic amino acids on the musculoskeletal system using one or more autophagy-inducing amino acids.

Abstract

A method for enhancing the effects of one or more anabolic amino acids on the musculoskeletal system in an individual in need thereof can comprise administering to the individual in need thereof a composition comprising one or more anabolic amino acids, wherein the composition also comprises one or more amino acids that induce autophagy in a total amount effective to render the composition at least neutral with respect to autophagy. In another embodiment, a method for overcoming one or more negative effects of one or more anabolic amino acids by preventing degenerative processes associated with decreased autophagy comprises administering to an individual in need thereof a composition comprising one or more anabolic amino acids, wherein the composition further comprises one or more amino acids that induce autophagy in a total amount effective to render the composition at least neutral with respect to autophagy.

Description

Background Art

[0001]

[0001] The present disclosure generally relates to compositions and methods that use one or more amino acids that induce autophagy, such as glycine, cysteine, proline, glutamic acid, valine, tyrosine, or precursors thereof, to enhance the effect of one or more anabolic amino acids such as leucine, isoleucine, arginine, glutamine, or citrulline. More specifically, the present disclosure relates to administering a composition comprising one or more anabolic amino acids, wherein the composition further comprises one or more autophagy-inducing amino acids or precursors thereof in an amount effective to render the composition at least neutral with respect to autophagy, and preferably positive with respect to autophagy, even if the one or more anabolic amino acids have a negative effect on autophagy. The composition can simultaneously promote protein synthesis and the removal of damaged cellular material. Recipients of administration can be, for example, critically ill patients such as patients in an intensive care unit (ICU); elderly patients, for example aged individuals; patients with sarcopenia or frailty; or individuals with chronic kidney disease (e.g., amino acid loss due to dialysis) and / or individuals with acute kidney injury.

[0002]

[0002] Great advances in intensive care have allowed critically ill patients to often survive acute illnesses that were previously fatal. However, for patients who survive this initial phase but enter a severe chronic phase, mortality remains high. Death is often due to refractory multiple organ failure, acute kidney injury and acute renal failure, severe myopathy, or less severe muscle weakness. Therapies such as nutrition, growth hormone, or androgens have been introduced to improve muscular myopathy and weakness, but these interventions have failed because they unexpectedly increase the risk of organ failure and death. Furthermore, nutritional supplementation for trauma and surgical patients can actually have harmful effects.

[0003]

[0003] No effective means for providing appropriate treatment and adequate nutrition to critically ill patients currently exists.

[0004]

[0004] Furthermore, while age-related decline in muscle mass and muscle function is unavoidable for all individuals, its progression varies greatly depending on genetic factors as well as environmental factors such as physical activity and nutritional intake. Sarcopenia is defined as a condition in which age-related decline in muscle mass and muscle function worsens to the point of affecting quality of life. In contrast, frailty is another classification of age-related decline in physical function, characterized by a decline in muscle strength and function rather than muscle mass. Sarcopenia is clinically defined by a decline in muscle mass and muscle function using divisions that stratify the aging population for individuals with pathological mobility. Future physical disability and mortality are predicted from sarcopenia, and sarcopenia was assigned an official ICD-10 disease code in 2016 (Anker et al., 2016). [Overview of the project] [Problems that the invention aims to solve]

[0005]

[0005] The degradation of cytoplasmic proteins is mediated by a cellular process called macroautophagy (also simply called autophagy). The autophagy process is also involved in inflammatory responses and promotes bacterial disruption of the immune system. Autophagy constitutes a major lysosomal degradation pathway that reuses damaged and sometimes harmful cellular materials, such as damaged mitochondria. In particular, autophagy mitigates the effects of cell death and extends lifespan in various aging models.

[0006]

[0006] As detailed in the experimental data later described herein, the inventors have found that several amino acids known to be involved in musculoskeletal anabolism via the mTOR pathway, such as arginine, glutamine, and leucine, and / or anabolic branched-chain amino acids, such as leucine and isoleucine, reduce the basal level of autophagy. However, the inventors have found that several amino acids, such as glycine, cysteine, proline, glutamic acid, valine, tyrosine, or their precursors, such as serine (as a precursor of glycine), N-acetylcysteine, and methionine (as a precursor of cysteine), can potently induce autophagy and thus counteract the negative effects that anabolic amino acids have on autophagy, such as muscle and neuromuscular degeneration, as well as decreased muscle mass and muscle function. [Means for solving the problem]

[0007]

[0007] Accordingly, in general embodiments, the present disclosure provides a method for enhancing the effects of one or more anabolic amino acids on the musculoskeletal system in an individual that requires enhanced effects on the musculoskeletal system. The method comprises administering to an individual a composition comprising one or more anabolic amino acids, the composition further comprising one or more autophagy-inducing amino acids or their precursors in a total amount effective such that the composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy.

[0008]

[0008] In one embodiment, one or more anabolic amino acids are selected from the group consisting of leucine, isoleucine, arginine, glutamine, citrulline, and mixtures thereof. One or more anabolic amino acids may contain at least one of leucine, glutamine, or arginine in an amount effective to activate mTOR in an individual.

[0009]

[0009] In one embodiment, one or more amino acids that induce autophagy are selected from the group consisting of glycine, cysteine, proline, glutamic acid, valine, tyrosine, or precursors thereof, and mixtures thereof.

[0010]

[0010] In one embodiment, the composition induces autophagy in skeletal muscle.

[0011]

[0011] In one embodiment, the individual is an elderly individual.

[0012]

[0012] In one embodiment, the individual has sarcopenia or frailty, or is at risk of developing sarcopenia or frailty.

[0013]

[0013] In one embodiment, the individual is severely ill.

[0014]

[0014] In one embodiment, the individual has severe myopathy or is at risk of developing severe myopathy.

[0015]

[0015] In one embodiment, the individual is in a severe condition accompanied by acute renal failure, or is at risk of developing acute renal failure.

[0016]

[0016] In one embodiment, the individual has a chronic kidney disease which may or may not be accompanied by a decrease in associated muscle mass or muscle function.

[0017]

[0017] In one embodiment, the individual has cachexia or muscle wasting secondary to a chronic disease such as cancer, chronic obstructive pulmonary disease (COPD), chronic heart failure (CHF), acute kidney disease, or chronic kidney disease (CKD).

[0018]

[0018] In one embodiment, the composition comprises a protein that provides at least a portion of one or more anabolic amino acids and / or at least a portion of one or more amino acids or their precursors that induce autophagy, wherein the protein is selected from the group consisting of (i) proteins derived from animal materials, (ii) proteins derived from plant materials, and (iii) mixtures thereof, and is preferably one or more of (a) milk protein, (b) whey protein, (c) casein salt, (d) casein micelle, (e) pea protein, (f) soy protein, and (g) mixtures thereof. In certain non-limiting examples, at least a portion of the protein is collagen, i.e., unhydrolyzed collagen and / or hydrolyzed collagen.

[0019]

[0019] The protein may be selected from the group consisting of (i) free amino acids, (ii) unhydrolyzed proteins, (iii) partially hydrolyzed proteins, (iv) extensively hydrolyzed proteins, and (v) mixtures thereof. The protein may contain peptides having a length of 2 to 10 amino acids. Optionally, at least a portion of the protein is 5 to 95% hydrolyzed. Optionally, the protein may have a formulation selected from the group consisting of (i) formulations in which at least 50% of the protein has a molecular weight of 1 to 5 kDa, (ii) formulations in which at least 50% of the protein has a molecular weight of 5 to 10 kDa, and (iii) formulations in which at least 50% of the protein has a molecular weight of 10 to 20 kDa.

[0020]

[0020] In one embodiment, the composition comprises a carbohydrate source and / or a fat source.

[0021]

[0021] In one embodiment, the administration step uses at least one route selected from the group of oral, enteral, parenteral, and intravenous injection.

[0022]

[0022] In one embodiment, the total amount of one or more anabolic amino acids is approximately equal to or greater than the total amount of one or more anabolic amino acids in the composition.

[0023]

[0023] In another embodiment, the Disclosure provides a composition comprising one or more anabolic amino acids, the composition further comprising one or more amino acids or their precursors that induce autophagy, in a total amount effective in such a way that the composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy. The composition may be selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional preparations that are reconstituted with water or milk before ingestion, food additives, pharmaceuticals, drinks, and combinations thereof.

[0024]

[0024] In another embodiment, the present disclosure provides a method for preparing a therapeutic composition, the method comprising the step of adding one or more autophagy-inducing amino acids or their precursors to a base composition comprising one or more anabolic amino acids to form a therapeutic composition, wherein the one or more autophagy-inducing amino acids or their precursors are added to the base composition in an amount effective such that the therapeutic composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy. The base composition and / or therapeutic composition may be formulated for administration by at least one route selected from the group of oral, enteral, parenteral, and intravenous injection. The base composition may be negative with respect to autophagy induction (i.e., the composition is negative overall), and / or the base composition may be neutral or positive with respect to autophagy induction, but may contain one or more anabolic amino acids in an amount that is negative with respect to autophagy induction (i.e., one or more anabolic amino acids form a negative portion with respect to autophagy induction).

[0025]

[0025] In one embodiment, the method comprises: quantifying the total amount of one or more anabolic amino acids in a base composition; and determining, using the total amount of one or more anabolic amino acids in the base composition, the total amount of one or more autophagy-inducing amino acids to be added to the base composition. The step of determining the total amount of one or more autophagy-inducing amino acids to be added to the base composition using the total amount of one or more anabolic amino acids in the base composition preferably comprises determining the expected effect on autophagy from the total amount of one or more anabolic amino acids. The step of determining the total amount of one or more autophagy-inducing amino acids to be added to the base composition using the total amount of one or more anabolic amino acids in the base composition is preferably carried out such that the expected effect on autophagy from the total amount of one or more autophagy-inducing amino acids added to the base composition is substantially equal to or greater than the expected effect on autophagy from the total amount of one or more anabolic amino acids, and further comprises determining the total amount of one or more autophagy-inducing amino acids to be added to the base composition.

[0026]

[0026] In another embodiment, the present disclosure provides a method comprising the step of administering to an individual in need thereof a composition that simultaneously promotes protein synthesis and removal of damaged cellular material, wherein the composition comprises one or more anabolic amino acids, and the composition further comprises one or more autophagy-inducing amino acids or precursors thereof in a total amount effective to allow the composition to be at least neutral with respect to autophagy, preferably positive with respect to autophagy.

[0027]

[0027] In another embodiment, the present disclosure provides a method of overcoming one or more negative effects of one or more anabolic amino acids by preventing degenerative processes associated with decreased autophagy. The method comprises the step of administering to an individual a composition comprising one or more anabolic amino acids, wherein the composition further comprises one or more amino acids that induce autophagy or precursors thereof, in an amount effective such that the composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy, for example in muscle.

[0028]

[0028] An advantage of one or more embodiments provided by the present disclosure is improving the condition of severely ill animals, severely ill humans, aged animals or aged humans.

[0029]

[0029] Another advantage of one or more embodiments provided by the present disclosure is preventing or treating excessive catabolism, for example in critically ill patients or aged individuals.

[0030]

[0030] Yet another advantage of one or more embodiments provided by the present disclosure is reducing or preventing the risk of morbidity or mortality due to excessive catabolism, for example in critically ill patients or aged individuals.

[0031]

[0031] An additional advantage of the present disclosure is recovering from, treating or curing multiple organ dysfunction syndrome in critically ill patients.

[0032]

[0032] An additional advantage of one or more embodiments provided by the present disclosure is protecting aged individuals from neurological diseases such as mild cognitive impairment, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, multiple sclerosis, Huntington's disease, dementia, and related rare neurological diseases.

[0033]

[0033] An additional advantage of one or more embodiments provided by the present disclosure is to protect aging individuals from muscle dysfunction such as sarcopenia, frailty, inclusion body myositis, drug-induced myopathy / muscle lysis such as corticosteroids or statins, and muscle wasting induced by immobility or hospitalization.

[0034]

[0034] An additional benefit of one or more embodiments provided by the present disclosure is the protection of patients with genetic disorders, including but not limited to muscular dystrophy such as Duchenne muscular dystrophy or type VI collagenous muscular dystrophy, mitochondrial encephalomyopathy, mitochondrial myopathy, glycogen storage disease, lysosomal storage disease, and Pompe disease.

[0035]

[0035] Another advantage of one or more embodiments provided by the present disclosure is that they are compositions that can be administered parenterally or enterally to critically ill patients, for example, as aqueous liquid compositions, in order to induce autophagy for the treatment of multi-organ dysfunction or burns.

[0036]

[0036] Yet another advantage of one or more embodiments provided by the present disclosure is to shorten the time that critically ill patients are on a ventilator or to shorten the time until they are removed from a ventilator.

[0037]

[0037] Another advantage of one or more embodiments provided by the present disclosure is to protect critically ill patients receiving parenteral nutrition from multiple organ failure or muscle weakness caused by parenteral nutrition delivery, particularly by unharmonious nutrition or relative overnutrition.

[0038]

[0038] An additional advantage of one or more embodiments provided by this disclosure is to protect aging individuals from muscle weakness.

[0039]

[0039] Yet another advantage of one or more embodiments provided by this disclosure is to improve the survival of critically ill patients or elderly individuals.

[0040]

[0040] An additional advantage of one or more embodiments provided by this disclosure is to accelerate the recovery of mobility or shorten the period of bed rest after discharge from the intensive care unit.

[0041]

[0041] Yet another advantage of one or more embodiments provided by this disclosure is that the effects are beneficial even when the critically ill patient has already progressed to a stage where his condition is life-threatening.

[0042]

[0042] Additional features and advantages will be described in the following detailed description of the invention and in the drawings, and will become apparent therefrom. [Brief explanation of the drawing]

[0043]

[0043] [Figure 1] This specification shows the activation of autophagy by individual amino acids in experimental examples disclosed herein. [Figure 2] This specification shows the activation of autophagy by individual amino acids in experimental examples disclosed herein. [Figure 3] This specification shows the activation of autophagy by individual amino acids in experimental examples disclosed herein. [Figure 4] This study describes the effect of a specific amino acid mixture (AA) on the force frequency response in aged rats and compares it to that of adult mice fed a standard diet. 20-month-old rats were fed either a control diet or the same diet supplemented with the specific amino acid mixture for 3 months. 6-month-old adult mice were fed a control diet for 3 months. Results are expressed as mean ± standard error. Adult: 6-month-old mice fed the control diet. Aged: 20-month-old mice fed the control diet. Aged+AA: 20-month-old mice fed the control diet supplemented with the specific amino acid mixture. [Modes for carrying out the invention]

[0044]

[0045] definition

[0046] The following are some definitions. However, definitions may also be found in the "Embodiments" section below, and the above heading "Definitions" does not mean that such disclosures in the "Embodiments" section are not definitions.

[0045]

[0047] All percentages are given by weight relative to the total weight of the composition unless otherwise specified. Similarly, all quantities and ratios are given by weight unless otherwise specified. Where a reference to pH is given, the value corresponds to the pH measured at 25°C using a standard apparatus. As used herein, “about,” “approximately,” and “substantially” are understood to refer to numbers within a numerical range, for example, within -10% to +10% of the reference figure, preferably within -5% to +5%, more preferably within -1% to +1%, and most preferably within -0.1% to +0.1% of the reference figure.

[0046]

[0048] Furthermore, all numerical ranges in this specification should be understood to include all integers, whole numbers, or fractions within that range. Moreover, these numerical ranges should be interpreted as supporting claims that cover any number or subset of a number within that range. For example, a disclosure of 1 to 10 should be interpreted as supporting ranges such as 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, and 3.5 to 9.9.

[0047]

[0049] Where used herein and in the appended claims, singular words include plural forms unless the context clearly indicates otherwise. Thus, references to “one,” “a,” and “the” (“a,” “an,” and “the”) generally include the plural forms of the respective terms. For example, a reference to “a certain amino acid” or “the amino acid” includes multiple such “amino acids.” The term “and / or” used in the context of “X and / or Y” should be interpreted as “X” or “Y” or “X and Y.” Similarly, “at least one of X or Y” should be interpreted as “X” or “Y” or “both X and Y.”

[0048]

[0050] Similarly, the terms “comprise,” “comprises,” and “comprising” should be interpreted as non-exclusive and potentially encompassing others. Likewise, the terms “include,” “including,” and “or” should all be interpreted as potentially encompassing others unless such interpretation is clearly prevented by the context. However, embodiments provided herein may not include any elements not specifically disclosed herein. Therefore, disclosures of embodiments defined using the term “comprising” are also disclosures of embodiments that “essentially consist of” and “consist of” the disclosed components.

[0049]

[0051] As used herein, the term “example,” especially when followed by a list of terms, is merely illustrative and descriptive, and should not be considered exclusive or comprehensive. All embodiments disclosed herein can be combined with any other embodiments disclosed herein unless otherwise expressly indicated.

[0050]

[0052] The term "animal" includes, but is not limited to, rodents, aquatic mammals, domestic animals such as dogs and cats, livestock such as sheep, pigs, cattle and horses, and mammals including, but not limited to, humans. Where "animal," "mammal," or their plural forms are used, these terms also apply to any animal capable of the effect indicated or intended to be indicated by the context of the text, such as an animal capable of autophagy. As used herein, the term "patient" is understood to include an animal receiving or scheduled to receive the treatment defined herein, such as a mammal, preferably a human. As used herein, the terms "individual" and "patient" are often used to refer to a human, but this disclosure is not limited to humans.

[0051]

[0053] Accordingly, the terms “individual” and “patient” refer to any animal, mammal, or human in which the methods and compositions disclosed herein may be beneficial. In fact, non-human animals experience long-term, serious illnesses that closely resemble those of humans. These severely ill animals experience the same metabolic disorders, immune disorders, and endocrine gland disorders, as well as the development of organ failure and muscle wasting, as corresponding to those in humans. Furthermore, animals also experience the effects of aging.

[0052]

[0054] The term "old age," in relation to humans, means an age of at least 55 years, preferably over 63 years, more preferably over 65 years, and most preferably over 70 years. The term "older adult" or "aging individual" in a human context means an age of at least 45 years, preferably over 50 years, and more preferably over 55 years, and includes elderly individuals.

[0053]

[0055] For other animals, “elderly adult” or “senior individual” means that the individual is more than 50% of the average lifespan of its species and / or genus within that species. An animal is considered “senior” when it is more than 66% of its average expected lifespan, preferably more than 75%, and more preferably more than 80%. An elderly cat or dog is at least about 5 years old. A senior cat or dog is at least about 7 years old.

[0054]

[0056] Sarcopenia is defined as a decline in muscle mass and muscle function (including muscle strength and walking speed) associated with aging. Sarcopenia can be assessed by one or more of the following: decreased muscle mass, decreased muscle strength, and decreased physical performance.

[0055]

[0057] Muscle mass loss can generally be measured by a decrease in the ALM index normalized to the square of height, specifically an ALM index of less than 7.00 kg / m2 for men and less than 5.40 kg / m2 for women. Physical performance decline can generally be measured by walking speed, specifically a walking speed of less than 0.8 m / sec. Muscle strength loss can generally be measured by grip strength, specifically a grip strength of less than 26 kg for men and less than 18 kg for women.

[0056]

[0058] Additionally or alternatively, sarcopenia can be diagnosed in subjects based on the definition of EWGSOP (European Working Group for Sarcopenia in Older People), as described, for example, by Crutz-Jentoft et al. (2010). Muscle mass loss can generally be judged by a decrease in the ALM index normalized to the square of height, specifically an ALM index of less than 7.23 kg / m2 for men and less than 5.67 kg / m2 for women. Physical performance loss can generally be judged by walking speed, specifically a walking speed of less than 0.8 m / sec. Muscle weakness can generally be judged by a decrease in grip strength, specifically a grip strength of less than 30 kg for men and less than 20 kg for women.

[0057]

[0059] Additionally or alternatively, sarcopenia can be diagnosed in subjects based on the Foundation for the National Institutes of Health (FNIH) definition, as described, for example, by Studenski et al. (2014). Muscle loss can generally be judged by a decrease in lean limb mass (ALM) normalized to body mass index (BMI; kg / m2), specifically by an ALM of less than 0.789 for men and less than 0.512 for women. Reduced physical performance can generally be judged by a decrease in walking speed, specifically a walking speed of less than 0.8 m / sec. Muscle weakness can generally be judged by a decrease in grip strength, specifically by a grip strength of less than 26 kg for men and less than 16 kg for women. Muscle weakness can also generally be judged by a decrease in grip strength-to-BMI ratio, specifically by a grip strength-to-BMI ratio of less than 1.00 for men and less than 0.56 for women.

[0058]

[0060] As used herein, “frailty” is defined as a clinically recognizable condition in which increased vulnerability results from a decline in reserve capacity and function across multiple physiological systems that occurs with aging, impairing the ability to cope with everyday or acute stressors. Where there is no established quantitative standard, frailty is functionally defined by Fried et al. as meeting three of the following five phenotypic criteria indicating impaired energy characteristics: (1) muscle weakness (baseline grip strength within the bottom 20% of the population, adjusted for sex and body mass index), (2) insufficient endurance and energy deficiency (self-reported fatigue related to maximal VO2), (3) bradykinesia (baseline being in the bottom 20% of the population based on 15-foot walking time, adjusted for sex and height), (4) reduced physical activity (weighted score of kilocalories expended per week at baseline, lowest quintile of physical activity identified for each sex; e.g., less than 383 kcal / week for men, 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). Pre-frailty stages where one or two of these criteria are present are identified as having a high risk of progressing to frailty.

[0059]

[0061] Cachexia is a complex metabolic syndrome associated with an underlying disease, characterized by muscle loss, sometimes accompanied by fat loss. Prominent clinical features of cachexia include (adjusted for fluid retention) weight loss in adults or growth retardation in children (excluding endocrine disorders).

[0060]

[0062] Cachexia is often seen in patients with diseases such as cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, cirrhosis, anorexia nervosa, chronic pancreatitis, and / or metabolic acidosis and neurodegenerative diseases.

[0061]

[0063] Cachexia is particularly common in certain types of cancer, such as pancreatic cancer, esophageal cancer, stomach cancer, intestinal cancer, lung cancer, and / or liver cancer.

[0062]

[0064] The internationally recognized diagnostic criteria for cachexia are weight loss exceeding 5% over a limited period, for example, 6 months, or current weight and height (Body Mass Index [BMI] of 20 kg / m²). 2 This refers to weight loss of more than 2% in individuals that are already showing wasting (less than 2%) or skeletal muscle mass (measured by DXA, MRI, CT, or bioelectrical impedance). Cachexia can progress gradually through various stages, namely from pre-cachexia to cachexia and then to refractory cachexia. Severity can be classified in combination with ongoing weight loss, depending on the degree of energy storage and body protein (BMI) depletion.

[0063]

[0065] In particular, cancer cachexia is characterized by a weight loss of more than 5% in the past six months (without simple starvation); or a BMI of less than 20 and a weight loss of more than 2%; or a lean body mass of the limbs that is not inconsistent with muscle loss (7.26 kg / m² for men). 2 Less than 5.45 kg / m² for women. 2 It is defined as being less than 2% in weight loss, and any weight loss being greater than 2% (Fearon et al., 2011).

[0064]

[0066] "Pre-cachexia" can be defined as a weight loss of 5% or less, in conjunction with anorexia nervosa and metabolic changes. Currently, there are no robust biomarkers to identify patients with pre-cachexia that are likely to progress further, or the rate at which pre-cachexia progresses. Refractory cachexia is defined essentially based on the patient's clinical characteristics and circumstances.

[0065]

[0067] The terms “treatment” and “treating” include any effect that results in improvement of a condition or disease (disorder), such as alleviating, reducing, controlling, or eliminating the condition or disease. This term does not necessarily mean that the subject is treated until a complete cure is achieved. Non-limiting examples of “treating” a condition or disease or “treating it” include (1) inhibiting the condition or disease, i.e., preventing the onset of the condition or disease or its clinical symptoms, and (2) alleviating the condition or disease, i.e., causing the temporary or permanent disappearance of the condition or disease or its clinical symptoms. Treatment may be related to the patient or to the physician.

[0066]

[0068] The term “prevention” or “prevention” means preventing the development of the clinical symptoms of the condition or disorder in an individual who may be exposed to or prone to a condition or disorder, but who has not yet exhibited or shown symptoms of the condition or disorder. The terms “condition” and “disease / disorder” mean any disease, condition, symptom, or sign.

[0067]

[0069] Relative terms such as "improved," "increased," and "enhanced" refer to the effect of a composition containing both one or more anabolic amino acids and one or more amino acids that induce autophagy, compared to a composition that is identical to the original composition except for the absence of one or more autophagy-inducing amino acids or the absence of one or more autophagy-inducing amino acids.

[0068]

[0070] The terms “food,” “food product,” and “food composition” mean products or compositions intended for consumption by an individual, such as a human, and providing to such individual at least one nutrient. The compositions of this disclosure, including many embodiments described herein, may include, be composed of, or essentially consist of, any additional or optional components, constituents, or restrictions described herein, or that are useful for dietary therapy, in addition to the essential elements and restrictions described herein.

[0069]

[0071] As used herein, “complete nutrition” contains sufficient variety and levels of major nutrients (proteins, lipids, and carbohydrates) and micronutrients to serve as the sole source of nutrition for the animal to which the composition is administered. An individual can obtain 100% of its nutritional needs from such a complete nutrition composition.

[0070]

[0072] As used herein, the term “critically ill patient” refers to an individual who is experiencing an acute, life-threatening condition, or who has been diagnosed as being at imminent risk of developing such a condition. Critically ill patients are medically unstable and, if left untreated, are likely to die (e.g., a mortality rate of >50%).

[0071]

[0073] Non-limiting examples of critically ill patients include patients with persistent or at risk of persistent life-threatening acute single or multiple organ failure due to disease or injury, patients who have undergone surgery and have complications, and patients who have undergone surgery on vital organs or major surgery in the past week.

[0072]

[0074] More specific, non-limiting examples of critically ill patients include patients with persistent or at risk of persistent life-threatening acute single or multiple organ failure due to disease or injury, and patients who have undergone surgery and have complications. Further specific, non-limiting examples of critically ill patients include patients requiring one or more of the following: cardiac surgery, cerebral surgery, thoracic surgery, abdominal surgery, vascular surgery, or transplantation, and patients suffering from one or more of the following: neurological disorders, traumatic brain injury, respiratory failure, peritonitis, multiple trauma, severe burns, or severe polyneuropathy.

[0073]

[0075] The term "Intensive Care Unit (ICU)" refers to a portion of a hospital that treats critically ill patients. The term "Intensive Care Unit" also includes nursing homes and clinics (e.g., private clinics) if the treatment activities performed there are the same as or similar to those of an ICU. "ICU patients" are encompassed within the term "critically ill patients."

[0074]

[0076] The term "multiple organ dysfunction" refers to a condition resulting from infection, reduced blood flow, increased metabolism, or injury such as an accident or surgery. "Multiple organ failure," which results in death in critically ill patients, is considered a descriptive clinical syndrome defined as dysfunction or failure of at least two vital organ systems. The most uniformly and specifically affected vital organ systems are the liver, kidneys, lungs, as well as the cardiovascular, nervous, and hematological systems. Non-limiting examples of multiple organ dysfunction include acute respiratory distress syndrome, heart failure, liver failure, kidney failure, respiratory failure, intensive care, shock, extensive burns, sepsis (e.g., systemic inflammatory response syndrome), and stroke.

[0075]

[0077] The term "enteral administration" includes oral administration (including oral forced nutrition) and rectal administration, but oral administration is preferred. The term "parenteral administration" refers to the delivery of a substance by a route other than the gastrointestinal tract, and includes intravenous, intra-arterial, intramuscular, intraventricular, intraosseous, intradermal, subarachnoid, and also intraperitoneal administration, intravesical instillation, and intracavernosal injection.

[0076]

[0078] The preferred parenteral administration is intravenous administration. A specific form of parenteral administration is the delivery of nutrients by intravenous administration. Parenteral nutrition is "total parenteral nutrition" when food is not administered by other routes. "Pareral nutrition" is preferably an isotonic or hypertonic aqueous solution (or a solid composition to be dissolved, or a liquid concentrate to be diluted to obtain an isotonic or hypertonic solution) containing sugars such as glucose, and further containing one or more of lipids, amino acids, and vitamins.

[0077]

[0079] Embodiment

[0080] One aspect of the present disclosure is a method for enhancing the effects of one or more anabolic amino acids on the musculoskeletal system in an individual requiring enhanced effects on the musculoskeletal system. The method comprises administering to an individual a composition comprising one or more anabolic amino acids, the composition further comprising one or more autophagy-inducing amino acids or their precursors in an amount effective in such a way that the composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy, for example, in muscle. The composition can be administered parenterally, enterally, or intravenously. The one or more anabolic amino acids are compounds different from the one or more autophagy-inducing amino acids.

[0078]

[0081] As used herein, “enhancing the effect on the musculoskeletal system” means that (i) the positive (anabolic) effect of one or more anabolic amino acids is greater than that of a composition identically formulated, except that one or more autophagy-inducing amino acids are absent or present in smaller amounts, and / or (ii) the positive (anabolic) effect of one or more anabolic amino acids lasts longer than that of a composition identically formulated, except that one or more autophagy-inducing amino acids are absent or present in smaller amounts.

[0079]

[0082] Non-limiting examples of suitable anabolic amino acids include leucine, isoleucine, arginine, glutamine, citrulline, and mixtures thereof. The composition may contain one or more of leucine, isoleucine, or arginine in free form and / or in the form of peptides and / or proteins (such as milk proteins, animal proteins, or plant proteins). Preferably, any leucine or arginine is present in the composition in an amount effective to activate mTOR. The daily dose of the composition may contain 0.175 to 142.85 mg / kg bw of leucine, preferably 0.35 to 71.425 mg / kg bw of leucine and 0.175 to 71.425 mg / kg bw of isoleucine; 20 to 300 mg / kg bw of arginine, preferably 50 to 200 mg / kg bw of arginine, and / or 20 to 300 mg / kg bw of citrulline, preferably 100 to 200 mg / kg bw of citrulline. The daily dose of one or more anabolic amino acids can be provided by taking one or more doses of the composition per day.

[0080]

[0083] Non-limiting examples of suitable amino acids for inducing autophagy include glycine, cysteine, proline, glutamic acid, valine, tyrosine, or their precursors, such as serine (as a precursor of glycine), N-acetylcysteine, methionine (as a precursor of cysteine), and mixtures thereof. The composition contains one or more autophagy-inducing amino acids or their precursors in an amount effective enough to make the composition at least neutral with respect to autophagy, and preferably positive with respect to autophagy, even if one or more anabolic amino acids have a negative effect on autophagy.

[0081]

[0084] Whey protein is rich in BCAAs such as leucine and isoleucine. Therefore, some embodiments of the composition include whey protein, which provides at least a portion of the anabolic amino acids in the composition. Furthermore, hydrolyzed collagen is a glycine and proline-rich material, and therefore, some embodiments of the composition include autophagy-inducing amino acids, which provide at least a portion of the anabolic amino acids in the composition.

[0082]

[0085] This composition can induce autophagy in muscles, such as skeletal muscle. Non-limiting examples of such muscles include one or more of the following: vastus lateralis, gastrocnemius, tibialis, soleus, extensor digitorum longus (EDL), biceps femoris, semitendinosus, semimembranosus, gluteus maximus, extraocular muscles, facial muscles, or diaphragm.

[0083]

[0086] Individuals requiring autophagy induction may be aged individuals such as aged animals or aged humans. In some embodiments, individuals requiring autophagy induction are aged animals or aged humans.

[0084]

[0087] Individuals requiring autophagy induction may be critically ill patients. In various embodiments, this method can treat or prevent multi-organ dysfunction in critically ill patients, for example, when homeostasis is impaired or disrupted due to parenteral nutrition; protect critically ill patients from multi-organ dysfunction; treat or prevent the development of lactic acidosis, for example, lactic acidosis induced by parenteral nutrition; treat or prevent muscle weakness in critically ill patients; reduce or prevent morbidity or death due to nutritional deterioration caused by parenteral nutrition; and / or prevent the collapse of bodily systems.

[0085]

[0088] In some embodiments, critically ill patients have at least one life-threatening condition selected from the group consisting of lactic acidosis, muscle weakness, hyperglycemia, multiple organ failure, homeostasis failure, and homeostasis disturbance. In one embodiment, critically ill patients have a non-infectious disease. In one embodiment, critically ill patients have multiple organ dysfunction that is not caused by or related to sepsis. Multiple organ dysfunction and muscle weakness are common in emergency settings and can be caused or aggravated by uncoordinated parenteral nutrition delivery or relative or absolute overnutrition delivered parenterally.

[0086]

[0089] In some embodiments, critically ill patients have at least one disease selected from the group consisting of severe trauma, multiple trauma, high-risk surgery, extensive surgery, cerebral trauma, cerebral hemorrhage, respiratory failure, peritonitis, acute kidney injury, acute liver injury, severe burns, severe polyneuropathy, severe myopathy, and ICU-acquired muscle weakness.

[0087]

[0090] In some embodiments, critically ill patients receive enteral or parenteral nutrition. In some embodiments, the composition treats or prevents mitochondrial dysfunction, for example, mitochondrial dysfunction induced by inadequate or uncoordinated parenteral nutrition to critically ill patients.

[0088]

[0091] In another aspect of the present disclosure, a method overcomes one or more negative effects of one or more anabolic amino acids by preventing denaturation processes associated with a decrease in autophagy. The method comprises administering a composition comprising one or more anabolic amino acids to an individual, wherein the composition further comprises one or more autophagy-inducing amino acids or their precursors in an amount effective such that the composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy, for example in muscle.

[0089]

[0092] As used herein, the term “protein” includes free amino acids, molecules of 2 to 20 amino acids (referred to herein as “peptides”), and also longer amino acid chains. Smaller peptides, i.e., chains of 2 to 10 amino acids, are suitable for the composition alone or in combination with other proteins. The “free form” of an amino acid is the monomeric form of the amino acid. Suitable amino acids include both natural and non-natural amino acids. The composition may contain one or more proteins, for example, one or more (i) peptides, (ii) longer amino acid chains, or (iii) mixtures of free amino acids; the mixture is preferably formulated to obtain a desired amino acid profile / content.

[0090]

[0093] The composition may contain a protein that provides at least a portion of one or more anabolic amino acids and / or at least a portion of one or more amino acids that induce autophagy, and at least a portion of the protein may be of animal or plant origin, and may be derived from dairy products such as milk protein, e.g., milk protein concentrate or milk protein isolate; casein salt or casein, e.g., casein micelle concentrate or casein micelle isolate; or whey protein, e.g., whey protein concentrate or whey protein isolate, one or more of which may be. Additionally or alternatively, at least a portion of the protein may be a plant protein such as one or more of soy protein or pea protein.

[0091]

[0094] Mixtures of these proteins, for example, a mixture in which casein is the majority but not the entirety of the protein, a mixture in which whey protein is the majority but not the entirety of the protein, a mixture in which pea protein is the majority but not the entirety of the protein, and a mixture in which soy protein is the majority but not the entirety of the protein, are also suitable. In one embodiment, at least 10% by weight, preferably at least 20% by weight, and more preferably at least 30% by weight of the protein is whey protein. In one embodiment, at least 10% by weight, preferably at least 20% by weight, and more preferably at least 30% by weight of the protein is casein. In one embodiment, at least 10% by weight, preferably at least 20% by weight, and more preferably at least 30% by weight of the protein is plant protein.

[0092]

[0095] Whey protein can be any whey protein selected from the group consisting of, for example, whey protein concentrate, whey protein isolate, whey protein micelle, whey protein hydrolysate, acid whey, sweet whey, denatured sweet whey (sweet whey from which caseinoglyco macropeptides have been removed), whey protein fractions, and any combination thereof.

[0093]

[0096] Casein can be obtained from any mammal, but preferably from milk, and preferably as casein micelles.

[0094]

[0097] The protein may not be hydrolyzed, may be partially hydrolyzed (i.e., peptides with a molecular weight of 3 kDa to 10 kDa and an average molecular weight of less than 5 kDa), or may be significantly hydrolyzed (i.e., 90% of peptides having a molecular weight of less than 3 kDa), for example, 5% to 95% hydrolyzed. In some embodiments, the peptide profile of the hydrolyzed protein may be within different molecular weight ranges. For example, the majority of the peptide (>50 mol% or >50 wt%) may have a molecular weight of 1 to 5 kDa, 5 to 10 kDa, or 10 to 20 kDa.

[0095]

[0098] In one embodiment, the composition includes a carbohydrate source. Any suitable carbohydrate may be used in the composition, including, but not limited to, starch (e.g., modified starch, amylose starch, tapioca starch, corn starch), sucrose, lactose, glucose, fructose, solid corn syrup, maltodextrin, xylitol, sorbitol, or a combination thereof.

[0096]

[0099] The carbohydrate source is preferably 50% or less of the energy of the composition, more preferably 36% or less of the energy of the composition, and most preferably 30% or less of the energy of the composition.

[0097]

[0100] In one embodiment, the composition includes a fat source. The fat source may include any suitable fat or fat mixture. Non-limiting examples of suitable fat sources include vegetable fats (e.g., olive oil, corn oil, sunflower oil, high-oleic sunflower oil, rapeseed oil, canola oil, hazelnut oil, soybean oil, palm oil, coconut oil, blackcurrant seed oil, borage oil, lecithin, etc.), animal fats (e.g., milk fat); or combinations thereof.

[0098]

[0101] The composition can be administered to individuals such as humans, for example, elderly individuals or critically ill individuals, in doses effective for treatment. The effective dose can be determined by those skilled in the art and may vary depending on several factors known to those skilled in the art, such as the severity of the condition and the individual's weight and overall condition.

[0099]

[0102] The composition is preferably administered to an individual at least two days a week, more preferably at least three days a week, most preferably seven days a week; for at least one week, at least one month, at least two months, at least three months, at least six months, or for a longer period. In some embodiments, the composition is administered to an individual continuously over several days, for example, until at least a therapeutic effect is achieved. In some embodiments, the composition can be administered to an individual daily for at least 30, 60, or 90 consecutive days.

[0100]

[0103] The above administration examples do not require uninterrupted daily administration. Rather, several short interruptions in administration, such as 2-4 day breaks during the administration period, may be permitted. The ideal duration of administration of this composition can be determined by those skilled in the art.

[0101]

[0104] In preferred embodiments, the composition is administered orally or enterally to an individual (e.g., via tube feeding). For example, the composition can be administered to an individual as a beverage, capsule, tablet, powder, or suspension.

[0102]

[0105] The composition may be any type of composition suitable for ingestion by humans and / or animals. For example, the composition may be selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional preparations that are reconstituted with water or milk before ingestion, food additives, pharmaceuticals, beverages and drinks. In one embodiment, the composition is an oral nutritional supplement (ONS), a complete nutritional formulation, a pharmaceutical, drug, or food product. In a preferred embodiment, the composition is administered to an individual as a beverage. The composition may be stored as a powder in a sachet and suspended in a liquid such as water for subsequent use.

[0103]

[0106] In some cases where oral or enteral administration is not possible or not recommended, this composition may also be administered parenterally.

[0104]

[0107] In some embodiments, the composition is administered to an individual in a single-dose dosage form, i.e., all compounds are present in a single formulation given to the individual with a meal. In other embodiments, the composition is administered simultaneously in separate dosage forms, for example, at least one component and one or more of the other components of the composition are administered simultaneously as separate components.

[0105]

[0108] In another embodiment, the Disclosure provides a method for preparing a therapeutic composition, comprising the step of adding one or more autophagy-inducing amino acids or their precursors to a base composition comprising one or more anabolic amino acids to form a therapeutic composition, wherein the one or more autophagy-inducing amino acids or their precursors are added to the base composition in an amount effective such that the therapeutic composition is at least neutral with respect to autophagy, preferably positive with respect to autophagy. The base composition and / or therapeutic composition may be formulated for administration by at least one route selected from the group of oral, enteral, parenteral, and intravenous injection. The base composition may be negative with respect to autophagy induction (i.e., the composition as a whole is negative), and / or the base composition may be neutral or positive with respect to autophagy induction, but may contain one or more anabolic amino acids in an amount that is negative with respect to autophagy induction (i.e., one or more anabolic amino acids form a portion that is negative with respect to autophagy induction).

[0106]

[0109] In one embodiment, the method includes the steps of: quantifying the total amount of one or more anabolic amino acids in the base composition; and determining the total amount of one or more amino acids added to the base composition to induce autophagy using the total amount of one or more anabolic amino acids in the base composition. The step of determining the total amount of one or more amino acids added to the base composition to induce autophagy using the total amount of one or more anabolic amino acids in the base composition preferably includes determining the effect on autophagy expected from the total amount of one or more anabolic amino acids. The step of determining the total amount of one or more amino acids added to the base composition to induce autophagy using the total amount of one or more anabolic amino acids in the base composition preferably further includes determining the total amount of one or more amino acids added to the base composition to induce autophagy such that the effect on autophagy expected from the total amount of one or more amino acids added to the base composition to induce autophagy is approximately equal to, or greater than, the effect on autophagy expected from the total amount of one or more anabolic amino acids. [Examples]

[0107]

[0110] Examples

[0111] The following non-limiting examples present scientific data that develop and support the concept of inducing autophagy in individuals requiring the composition by administering a composition containing one or more anabolic amino acids, wherein the composition further contains one or more autophagy-inducing amino acids in an amount effective enough to make the composition at least neutral with respect to autophagy, and preferably positive with respect to autophagy, even if one or more anabolic amino acids have a negative effect on autophagy.

[0108]

[0112] Example 1

[0113] Experimental protocol

[0114] The zebrafish autophagy reporter was generated by the stable expression of LC3 protein fused to ZsGreen under the control of a skeletal muscle-specific promoter. Larvae of outcrossed transgenic zebrafish were cultured at 28°C under standard laboratory conditions and treated in 96-well plates with the indicated amino acid at concentrations ranging from 0.25 to 10 mM for 48 hours post-fertilization (n=24). After 16 hours of treatment, the larvae were anesthetized with 0.016% tricaine and imaged at 20x magnification using an ImageXpress confocal system (Molecular Devices). Z-stack images were captured for each larva, and the maximum projection image was constructed. The number of LC3 granules was calculated using MetaXpress software (Molecular Devices) and normalized by the fish area.

[0115] Figures 1-3 show the experimental results.

[0109]

[0116] Example 2

[0117] Materials and methods

[0118] Aged mice are a good model for evaluating the effects of nutritional therapy on age-related decline. Using this model, age-related muscle function decline can be observed, similar to that in humans. In this model, 20-month-old mice were fed either a normal diet or the same diet supplemented with a specific amino acid mixture (AA) for 3 months (see Table 1). The dose of the AA mixture was 1.65 mg / kg BW / day.

[0119]

[0110] [Table 1]

[0111]

[0120] To evaluate the effects of aging, adult mice (6 months old) were also fed a normal diet for 3 months. The age-related decline in in vivo force-contraction frequency response was measured using a 305C muscle liver system. Figure 4 shows the effect of a specific amino acid mixture (AA) on the force-contraction frequency response in aged rats and a comparison with adult mice fed a normal diet. The force-contraction frequency response was significantly improved with AA supplementation, suggesting improved muscle function.

[0112]

[0121] It should be understood that various changes and modifications to the current 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 subject matter of the invention and without impairing the intended advantages. Therefore, such changes and modifications are intended to be covered by the appended claims.

Claims

1. A composition for use in a method to enhance the effects of one or more anabolic amino acids on the musculoskeletal system in individuals where it is necessary to enhance the effects on the musculoskeletal system, The method includes the step of administering the composition to the individual, The composition comprises lysine, the anabolic amino acid, and an amino acid that induces autophagy. The autophagy-inducing amino acids consist of proline, glycine, cysteine, and valine. A composition wherein the assimilated amino acids consist of leucine and isoleucine.

2. The composition according to claim 1, wherein the composition induces autophagy in skeletal muscle.

3. The composition according to claim 1, wherein the individual is an elderly individual.

4. The composition according to claim 1, wherein the individual has sarcopenia or frailty, or is at risk of developing sarcopenia or frailty.

5. The composition according to claim 1, wherein the individual has at least one condition selected from the group consisting of (i) severe, (ii) acute kidney injury, (iii) chronic kidney injury, (iv) decreased muscle mass due to chronic kidney disease, and (v) decreased muscle function due to chronic kidney disease.

6. The composition according to claim 1, wherein the individual has severe myopathy or is at risk of developing severe myopathy.

7. The composition according to claim 1, wherein the individual has cachexia or muscle wasting secondary to cancer, chronic obstructive pulmonary disease (COPD), chronic heart failure (CHF), acute kidney disease, or chronic kidney disease (CKD).

8. The composition according to claim 1, wherein the composition comprises a protein that provides at least a portion of the assimilated amino acids and / or at least a portion of the autophagy-inducing amino acids, wherein at least a portion of the protein is selected from the group consisting of (i) proteins derived from animal materials, (ii) proteins derived from plant materials, and (iii) mixtures thereof.

9. The composition according to claim 8, wherein the protein is selected from the group consisting of (i) milk protein, (ii) whey protein, (iii) casein salt, (iv) casein micelle, (v) pea protein, (vi) soy protein, and (vii) mixtures thereof.

10. The composition according to claim 8, wherein the protein is selected from the group consisting of (i) free amino acids, (ii) unhydrolyzed proteins, (iii) partially hydrolyzed proteins, (iv) extensively hydrolyzed proteins, and (v) mixtures thereof.

11. The composition according to claim 8, wherein at least a portion of the protein is hydrolyzed by 5 to 95%.

12. The composition according to claim 8, wherein the protein comprises a peptide having a length of 2 to 10 amino acids.

13. The composition according to claim 8, wherein the protein comprises a formulation selected from the group consisting of (i) a formulation in which at least 50% of the protein has a molecular weight of 1 to 5 kDa, (ii) a formulation in which at least 50% of the protein has a molecular weight of 5 to 10 kDa, and (iii) a formulation in which at least 50% of the protein has a molecular weight of 10 to 20 kDa.

14. The composition according to claim 1, wherein the composition comprises a carbohydrate source and / or a fat source.

15. The composition according to claim 1, wherein the administration step uses at least one route selected from the group consisting of oral, enteral, parenteral, and intravenous injection.

16. The composition according to claim 1, wherein the composition is selected from the group consisting of food compositions, dietary supplements, nutritional compositions, nutraceuticals, powdered nutritional preparations that are reconstituted with water or milk before ingestion, food additives, pharmaceuticals, drinks, and combinations thereof.

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