Compositions for promoting muscle growth and muscle function and methods for using the same

Abstract

To provide compositions for use in increasing muscle mass, preventing muscle atrophy, promoting muscle growth, and treating various other conditions and diseases, and methods for the treatment.SOLUTION: A composition for increasing muscle mass comprises: a. dileucine; b. leucine; and c. pharmaceutically acceptable carriers thereof. Disclosed compositions comprise dileucine and leucine and pharmaceutically acceptable salts thereof. The compositions are administered to subjects in need thereof to increase muscle mass, prevent muscle atrophy, and / or promote muscle growth.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - Reference to Related Applications

[0001] This application claims priority to U.S. Provisional Application No. 62 / 659,474, filed on April 18, 2018, and titled "COMPOSITIONS AND METHODS OF USE THEREOF TO PROMOTE MUSCLE GROWTH", which is hereby incorporated by reference in its entirety under 35 U.S.C. §119(e). AND METHODS OF USE THEREOF TO PROMOTE MUSCLE GROWTH” and is hereby incorporated by reference in its entirety under 35 U.S.C. §119(e).

Background Art

[0002]

[0002] An increase in muscle function is an important goal for professional athletes to fitness enthusiasts. Further, the maintenance of muscle function is highly important for healthy aging. The present disclosure relates to compositions and methods of using such compositions for increasing muscle mass in a subject or preventing muscle atrophy in a subject. The present disclosure further relates to compositions and methods of using such compositions for enhancing protein concentration or muscle mass in a mammal, and methods for enhancing protein concentration or muscle mass in a mammal. Further, the present disclosure relates to compositions and methods of using such compositions for increasing muscle mass in a subject or preventing muscle atrophy in a subject. The muscle atrophy - preventing compositions are significantly effective in preventing muscle mass decline and increasing muscle mass, and thus are useful in preventing and treating various muscle diseases, such as sarcopenia and disuse atrophy.

Summary of the Invention

[0003]

[0003] Described herein are compositions for increasing muscle mass, muscle function, and / or preventing muscle atrophy. In certain aspects, the compositions comprise dileucine, leucine, and their pharmaceutically acceptable carriers. In further aspects, dileucine is present in about 10% to 90% (w / w). In further aspects, dileucine is present in about 20% to 80% (w / w). In further aspects, dileucine is present in about 30% to 70% (w / w). In further aspects, dileucine is present in about 40% to 60% (w / w). In even further aspects, dileucine is present from about 50% (w / w).

[0004]

[0004] Various embodiments of compositions and methods for increasing muscle mass or strength and / or reducing or treating muscle atrophy are described herein. In particular: Disclosed is a method for increasing muscle mass and / or muscle strength in a subject by administering to a subject an effective amount of a composition comprising dileucine, trileucine, and Leu-Leu-R, where R is an amino acid or amino acid derivative, and at least one amino acid or peptide selected from pharmaceutically acceptable salts thereof. In particular, the compound is dileucine. In particular, the at least one amino acid or peptide comprises leucine and dileucine. In further aspects, dileucine is present in about 10% to about 90% (w / w). In further aspects, dileucine is present in about 30% to about 70% (w / w). In even further aspects, dileucine is present in about 50% (w / w).

[0005]

[0005] In a further aspect, the composition is a dileucine acetate salt. In a specific aspect, the composition is Leu-Leu-R, where R is a branched-chain amino acid. In a further aspect, R is an essential amino acid. In a further aspect, R is a conditionally essential amino acid selected from the group consisting of arginine, cysteine, glutamine, glycine, proline, and tyrosine. In an exemplary embodiment, R is tyrosine. In an alternative embodiment, R is a non-essential amino acid selected from the group consisting of alanine, aspartic acid, asparagine, glutamic acid, serine, selenocysteine, and pyrrolelysine. In a further alternative embodiment, R is an amino acid derivative selected from the list consisting of creatine, carnitine, creatinol, beta-alanine, taurine, and beta-hydroxy-beta-methylbutyrate.

[0006]

[0006] In various embodiments, administration of the composition to a subject synergistically increases plasma leucine levels compared to administration of a composition containing leucine but not dileucine. In certain aspects, administration of the composition to a subject synergistically increases muscle mass and / or muscle strength compared to administration of a composition containing leucine but not dileucine.

[0007]

[0007] Further disclosed herein is a method for preventing or treating muscle atrophy in a subject, comprising the step of administering to a subject an effective amount of a composition comprising dileucine, trileucine, and at least one more selected amino acid or peptide, Leu-Leu-R, where R is an amino acid or amino acid derivative. In a particular aspect, the composition comprises leucine and dileucine. In a particular aspect, the method is used to treat or prevent muscle atrophy resulting from sarcopenia. In a further aspect, muscle atrophy is a result of cachexia. In a still further aspect, muscle atrophy is a result of muscle fixation.

[0008]

[0008] In some aspects, dileucine is present at approximately 10% to 90% (w / w). In further aspects, dileucine is present at approximately 30% to 70% (w / w). In further aspects, dileucine is present at approximately 50% (w / w).

[0009]

[0009] In various aspects, administration of the composition to a subject synergistically increases plasma leucine levels compared to administration of a composition containing leucine but not dileucine. In specific aspects, the composition is administered in a therapeutically effective dose.

[0010]

[0010] A method for improving cognition or preventing age-related memory / cognitive loss in a subject is further disclosed herein, comprising the step of administering to the subject a therapeutically or prophylactically effective amount of a composition comprising a compound selected from the list comprising dileucine, trileucine, and Leu-Leu-R, where R is an amino acid or amino acid derivative. In certain aspects, administration of the composition increases BDNF levels. In further aspects, administration of the composition increases NGF. In even further aspects, the composition is administered in a neuroprotective amount.

[0011]

[0011] Compositions for the treatment of conditions comprising dileucine, leucine, and their pharmaceutically acceptable carriers are further disclosed herein. In particular aspects, conditions are at least one of obesity, immune system dysfunction, insulin secretion-related disorders, diabetes, pathogenicity-related conditions, cardiovascular disorders, heart disorders, degenerative diseases, sarcopenia, eye diseases, fibrous diseases, age-related disorders, improved skin hydration and collagen synthesis, liver diseases, and Crohn's disease.

[0012]

[0012] Numerous embodiments are disclosed, but illustrative embodiments of the disclosed apparatus, systems and methods are shown, and further embodiments of the disclosure will become apparent to those skilled in the art from the following detailed description. As will be understood, all disclosed apparatus, systems and methods are modifiable in various obvious aspects without departing from the spirit and scope of this disclosure. Accordingly, the figures and detailed description should be considered illustrative and non-restrictive. [Brief explanation of the drawing]

[0013] [Figure 1]

[0013] Figure 1 is a schematic diagram of the leucine processing pathway according to a specific embodiment. [Figure 2]

[0014] Figure 2 shows the plasma leucine concentration after administration of the disclosed composition according to a specific embodiment. [Figure 3]

[0015] Figure 3 shows the plasma isoleucine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 4]

[0016] Figure 4 shows the plasma valine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 5]

[0017] Figure 5 shows the total plasma BCAA concentration after administration of the disclosed composition according to a particular embodiment. [Figure 6]

[0018] Figure 6 shows the plasma threonine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 7]

[0019] Figure 7 shows the plasma methionine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 8]

[0020] Figure 8 shows the plasma tryptophan concentration after administration of the disclosed composition according to a particular embodiment. [Figure 9]

[0021] Figure 9 shows the plasma phenylalanine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 10]

[0022] Figure 10 shows the plasma lysine concentration after administration of the disclosed composition according to a particular embodiment. [Figure 11]

[0023] Figure 11 shows the total plasma essential amino acids (EAA) after administration of the disclosed composition according to a particular embodiment. [Figure 12]

[0024] Figure 12 shows a schematic diagram of an experimental protocol for one example. [Figure 13]

[0025] Figure 13 shows the leucine Tmax after administration of the disclosed composition according to a particular embodiment. [Figure 14]

[0026] Figure 14 shows the leucine AUC after administration of the disclosed composition according to a particular embodiment. [Figure 15]

[0027] Figure 15 shows a schematic diagram of an experimental protocol for one example. [Figure 16]

[0028] Figure 16 shows exemplary plasma muscle protein synthesis data from a muscle biopsy after administration of the disclosed composition according to a particular embodiment. **DETAILED DESCRIPTION OF THE INVENTION**

[0014]

[0029] Prior to the disclosure and description of the present compounds, compositions, articles, systems, devices, and / or methods, it is to be understood that these are not limited to a particular synthesis method or, unless otherwise specified, to particular reagents, as such can of course vary. It is also to be understood that the nomenclature used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, exemplary methods and materials are described herein.

[0015]

[0030] As used herein, the term "subject" refers to the target of administration, e.g., a subject. Thus, the subject of the methods disclosed herein can be a vertebrate, such as a mammal, fish, bird, reptile, or amphibian. Alternatively, the subject of the methods disclosed herein can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, it is intended to include both male and female subjects, as well as adult and neonatal subjects, and fetuses. In one aspect, the subject is a mammal. A patient refers to a subject afflicted with a disease or disorder. The term "patient" includes human and veterinary subjects. In some aspects of the disclosed methods, the subject has been diagnosed as having a need for treatment of one or more muscle disorders prior to the administration step. Some of the disclosed methods In some aspects of the disclosed method, the subject has been diagnosed as needing to increase muscle mass before the administration step.

[0016]

[0031] In this specification, the term “treatment” refers to the medical management of a patient with the intention of curing, alleviating, stabilizing, or preventing a disease, pathological condition, or disorder. This term includes active treatment, i.e., treatment specifically directed toward improvement of a disease, pathological condition, or disorder, and also causal treatment, i.e., treatment directed toward the elimination of the cause of the associated disease, pathological condition, or disorder. Furthermore, this term includes palliative treatment, i.e., treatment designed to alleviate symptoms rather than cure the disease, pathological condition, or disorder; preventive treatment, i.e., treatment directed toward minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, i.e., treatment used to support another specific therapy directed toward improvement of the associated disease, pathological condition, or disorder. In various aspects, the term encompasses any treatment of a subject, including mammals (e.g., humans), which includes: (i) preventing the onset of a disease in a subject who may be predisposed to the disease but has not yet been diagnosed with the disease; (ii) inhibiting the disease, i.e., suppressing its development; or (iii) alleviating the disease, i.e., causing a regression of the disease. In one aspect, the subject is a mammal, e.g., a primate, and in another aspect, the subject is a human. The term “subject” also includes domesticated animals (e.g., cats, dogs, etc.), livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mice, rabbits, rats, guinea pigs, fruit flies, etc.).

[0017]

[0032] In this specification, the terms “prevent” or “prevent” mean making something impossible, avoiding, preventing, escaping, stopping, or hindering something from happening, particularly through prior action. Wherever “reduce,” “hinder,” or “prevent” is used in this specification, unless otherwise specified, the use of the other two words is also understood to be explicitly disclosed.

[0018]

[0033] In this specification, the term “diagnosed” means that a person is subjected to a physical examination by a person skilled in the art, for example, a physician, and is found to have a condition that can be diagnosed or treated by the compounds, compositions, or methods disclosed herein. For example, “diagnosed with muscular atrophy” means that a person is subjected to a physical examination by a person skilled in the art, for example, a physician, and is found to have a condition that can be diagnosed or treated by a compound or composition that can increase muscle mass. As a further example, “diagnosed to need muscle mass increase” means that a person is subjected to a physical examination by a person skilled in the art, for example, a physician, and is found to have a condition characterized by muscular atrophy or another disease in which muscle mass increase would be beneficial to the subject. Such diagnoses may relate to disorders such as muscular atrophy, as discussed herein.

[0019]

[0034] In this specification, the phrase "identified as needing treatment for a disorder," etc., refers to the selection of a subject based on the need for treatment for a disorder. For example, a subject may be identified as needing treatment for a disorder (e.g., a disorder related to muscle atrophy) based on a prior diagnosis by a person skilled in the art, and then be subjected to treatment for the disorder. In one aspect, the identification is intended to be performed by a person different from the person who made the diagnosis. In a further aspect, the administration is also intended to be performed by a person who subsequently administers the treatment.

[0020]

[0035] In this specification, the terms “administering” and “administering” refer to any method of providing a pharmaceutical preparation to a subject. Such methods are well known to those skilled in the art, and This includes, but is not limited to, oral administration, transdermal administration, inhalation administration, nasal administration, topical administration, vaginal administration, ocular administration, ear administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injectable methods such as intravenous, intra-arterial, intramuscular, and subcutaneous administration. Administration may be continuous or intermittent. In various aspects, the preparation may be administered therapeutically; that is, to treat an existing disease or condition. In even more various aspects, the preparation may be administered prophylactically; that is, to prevent a disease or condition.

[0021]

[0036] While it is undesirable to be bound by any particular theory of mechanism of action, leucine levels within an organism are indicators of its physiological state, including how much food is available, how much insulin is about to be needed, and whether new muscle mass can be created. For example, leucine reduces proteolysis in humans, suggesting that leucine is a regulator of protein metabolism in humans. Furthermore, leucine stimulates Akt and protein synthesis, resulting in increased ATP demand. These increases in energy demand stimulate the activity and cellular expression levels of metabolic regulators, including AMPK, PPARβ / δ, and PGC-1α, leading to a simultaneous increase in oxidative metabolism, mitochondrial biosynthesis, and GLUT4 content. The increase in GLUT4 content then promotes increased glucose uptake, assisting the increased energy need. Increased energy uptake simultaneously promotes substrate oxidation and storage (partly through increased PPARγ), leading to an increase in cellular lipid content.

[0022]

[0037] As shown in Figure 1, sestrin 2 is central to leucine-mediated energy sensing and metabolic regulation processes. Sestrin 2 links cellular and systemic concentrations of leucine to the regulation of biological metabolism and growth. Therefore, when leucine binds to sestrin 2, this causes sestrin to be released from a complex containing the mTORC1 regulator GATOR2, and upon release, this molecule activates the mTORC1 complex. Thus, it is well recognized that regulating the activity of this leucine and mTORC-mediated process can provide novel strategies for the treatment of a wide range of conditions and diseases.

[0023]

[0038] The compositions disclosed herein are capable of influencing leucine concentration in living organisms, and the resulting mTORC activity is expected to have a wide range of therapeutic benefits. In various embodiments, compositions comprising leucine and dipeptides, including dileucine, are disclosed herein, which can substantially modify the control of leucine concentration in vivo and have a wide range of therapeutic benefits. In various aspects, the benefits of the compounds described herein may include the ability to increase muscle growth. In various other aspects, the disclosed compositions may be used to support muscle homeostasis, prevent and / or treat sarcopenia, and affect satiety.

[0024]

[0039] Compositions and methods for promoting muscle growth and / or preventing or treating muscle atrophy are disclosed herein. In certain aspects, the disclosed methods include the step of administering a composition to a subject, wherein the composition comprises the compound dileucine. Dileucine refers to a dipeptide composed of two L-leucine molecules. In certain aspects, dileucine has the structure:

[0025] [ka]

[0026] It may have the following characteristics. Dileucine may also be called L-leucyl-L-leucine or Leu-Leu, and has the CAS number 3303-31-9.

[0040] In certain aspects, the disclosure is a composition for increasing muscle mass, comprising dileucine, leucine, and their pharmaceutically acceptable carriers. In various aspects, dileucine is present at approximately 10% to 90% (w / w). In further aspects, dileucine is present at approximately 20% to 80% (w / w). In further aspects, dileucine is present at approximately 30% to 70% (w / w). In further aspects, dileucine is present at approximately 40% to 60% (w / w). In even further aspects, dileucine is present at approximately 50% (w / w).

[0027]

[0041] In an alternative configuration, dileucine is present in approximately 10% to 90% (w / w) proportions, and leucine in approximately 90% to 10% (w / w) proportions. In a further configuration, dileucine is present in approximately 20% to 80% (w / w) proportions, and leucine in approximately 80% to 20% (w / w) proportions. In a further configuration, dileucine is present in approximately 30% to 70% (w / w) proportions, and leucine in approximately 70% to 30% (w / w) proportions. In a further configuration, dileucine is present in approximately 40% to 60% (w / w) proportions, and leucine in approximately 60% to 40% (w / w) proportions. In a further configuration, dileucine is present in approximately 50% (w / w) proportions, and leucine in approximately 50% (w / w) proportions.

[0028]

[0042] In certain aspects, the disclosed method includes the step of administering a composition comprising a dileucine salt. In an exemplary embodiment, the composition has the following structure:

[0029] [ka]

[0030] It is a dileucine acetate salt that may have [a specific characteristic].

[0043] In certain aspects, the disclosed method comprises the step of administering a composition to a subject, wherein the composition comprises trileucine. Trileucine means a tripeptide comprising three L-leucine molecules. In certain aspects, trileucine has the structure:

[0031] [ka]

[0032] It holds.

[0044] Trileucine may also be called trileucine:LEU-LEU-LEU;H-LEU-LEU-LEU-OH;L-leucylleucylleucine;leucyl-leucyl-leucine;Leu-leu-leucrystalline;L-leucyl-L-leucyl-L-leucine;L-leucyl-L-leucyl-L-leucine;Leu-Leu-Leu-OH≧S)-2((S)-2-((S)-2-amino-4-methylpentanamide)-4-methylpentanamide)-4-methylpentanoic acid.

[0033]

[0045] In a further aspect, the disclosed method comprises administering a composition to a subject, wherein the composition comprises a tripeptide comprising two L-leucine units and one amino acid or amino acid derivative. In a particular embodiment, the amino acid is selected from the branched-chain amino acid (BCAA) group, including, but not limited to, isoleucine, leucine, and valine. In a further embodiment, the amino acid is selected from the essential amino acid group, including, but not limited to, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. In a still further embodiment, the amino acid is selected from the conditionally essential amino acid group, including, but not limited to, arginine, cysteine, glutamine, glycine, proline, and tyrosine. In a particular embodiment, the conditionally essential amino acid is tyrosine. In an exemplary embodiment, the composition has the structure:

[0034] [ka]

[0035] It contains dileucine and tyrosine.

[0046] In a further embodiment, the amino acid is selected from the group of non-essential amino acids, including, but not limited to, alanine, aspartic acid, asparagine, glutamic acid, serine, selenocysteine, and pyrrolsine. In a further embodiment, the amino acid derivative is creatine, carnitine, creatinine The following are selected from the group: ru, beta-alanine, taurine, and beta-hydroxy-beta-methylbutyrate.

[0036]

[0047] In certain aspects, the disclosure is a method for increasing muscle mass and / or muscle strength in a subject, comprising the step of administering to the subject an effective amount of a composition comprising at least one amino acid or peptide selected from dileucine, trileucine, and Leu-Leu-R, where R- is an amino acid or amino acid derivative, and a pharmaceutically acceptable salt thereof. In various aspects, the at least one amino acid or peptide comprises leucine and dileucine and / or a pharmaceutically acceptable salt thereof.

[0037]

[0048] In certain aspects, dileucine is present at approximately 10% (w / w) to 90% (w / w). In further aspects, dileucine is present at approximately 30% to 70% (w / w). In yet another aspect, dileucine is present at approximately 50% (w / w).

[0038]

[0049] In various alternative forms, dileucine is present in approximately 10% to 90% (w / w) proportions, and leucine in approximately 90% to 10% (w / w) proportions. In further forms, dileucine is present in approximately 30% to 70% (w / w) proportions, and leucine in approximately 70% to 30% (w / w) proportions. In yet another form, dileucine is present in approximately 50% (w / w) proportions, and leucine in approximately 50% (w / w) proportions.

[0039]

[0050] In these and other embodiments, administration of the composition to a subject synergistically increases plasma leucine levels compared to administration of a composition containing leucine but not dileucine. Furthermore, administration of the composition to a subject synergistically increases muscle mass and / or muscle strength compared to administration of a composition containing leucine but not dileucine.

[0040]

[0051] In various aspects, the composition contains at least about 95% dileucine; and between about 0.1% and 5% trileucine, and pharmaceutically acceptable salts thereof. In further aspects, trileucine is present in amounts between about 0.1% and 3%, and the composition further contains about 0.1% and 2% tetraleucine. In yet another aspect, trileucine is present in amounts of about 0.4%, and tetraleucine is present in amounts of about 0.2%.

[0041]

[0052] According to certain alternative embodiments, the composition comprises at least about 95% dileucine; and between about 0.1% and 5% tetraleucine, and pharmaceutically acceptable salts thereof.

[0042]

[0053] In certain aspects of the embodiments described above, the composition is substantially leucine-free.

[0054] Disclosed, in accordance with certain further aspects, a method for increasing muscle mass and / or muscle strength in a subject, comprising the step of administering to the subject an effective amount of a composition comprising at least about 95% dileucine; and between about 0.1% and 5% leucine, and a pharmaceutically acceptable salt thereof, in accordance with exemplary aspects of these embodiments.

[0043]

[0055] In certain aspects, the compositions administered according to the disclosed methods are produced through bacterial fermentation. In these embodiments, fermentation techniques are used, utilizing di / tri / tetrapeptide-forming enzymes that directly link amino acids, followed by an extraction process.

[0044]

[0056] This specification discloses a method for promoting muscle growth through the administration of an effective amount of one or more compositions disclosed herein, in a particular aspect. Therefore, administration of an effective dose of the disclosed composition results in higher levels of plasma leucine through improved delivery and / or transport. In a specific further aspect, administration of an effective dose of the disclosed composition results in higher levels of muscle protein synthesis per gram in the subject. In a further further aspect, administration of an effective dose of the disclosed composition results in faster transport to plasma and faster and more optimal MPS (muscle protein synthesis) in the subject. In a further further aspect, administration of an effective dose of the disclosed composition results in improved muscle adhesion in the subject.

[0045]

[0057] In certain embodiments, the compositions disclosed herein may be administered in combination with an intensity training regimen. As will be recognized by those skilled in the art, administration of an effective amount of the disclosed compositions results in improved intensity and improved exercise performance / ergogenesis in the subject.

[0046]

[0058] In one aspect, the disclosed compound inhibits muscle atrophy. In another aspect, the disclosed compound increases muscle mass. In yet another aspect, the disclosed compound induces muscle hypertrophy. In yet another aspect, the disclosed compound inhibits muscle atrophy and increases muscle mass. In yet another aspect, the disclosed compound inhibits muscle atrophy and induces muscle hypertrophy. In yet another aspect, the inhibition of muscle atrophy is observed in the subject. In yet another aspect, the increase in muscle mass is observed in the subject. In yet another aspect, the subject is a mammal. In yet another aspect, the mammal is a human.

[0047]

[0059] In certain aspects, administration of the disclosed composition is effective in preventing or treating age-related muscle atrophy or sarcopenia. In further aspects, administration of the disclosed composition is effective in preventing or treating muscle atrophy associated with muscle immobilization, such as that which frequently occurs with cast immobilization of fractured bones. In further aspects, administration of the disclosed composition is effective in preventing or treating muscle atrophy associated with disease, such as cancer, also known as cachexia.

[0048]

[0060] Disclosed herein is a method for preventing or treating muscle atrophy in a subject, comprising the step of administering an effective amount of the disclosed composition to the subject. In certain aspects, the composition comprises leucine and dileucine.

[0049]

[0061] In various forms, dileucine is present at approximately 10% (w / w) to 90% (w / w). In further forms, dileucine is present at approximately 30% to 70% (w / w). In yet another form, dileucine is present at approximately 50% (w / w).

[0050]

[0062] In various alternative forms, dileucine is present in approximately 10% to 90% (w / w) proportions, and leucine in approximately 90% to 10% (w / w) proportions. In further forms, dileucine is present in approximately 30% to 70% (w / w) proportions, and leucine in approximately 70% to 30% (w / w) proportions. In yet another form, dileucine is present in approximately 50% (w / w) proportions, and leucine in approximately 50% (w / w) proportions.

[0051]

[0063] The composition is administered to subjects with sarcopenia according to specific aspects. The composition is administered in therapeutically effective doses in various aspects. The composition is administered in prophylactically effective doses in further aspects (e.g., to subjects at risk of developing sarcopenia, cachexia, or fixation-induced atrophy).

[0052]

[0064] In certain embodiments, administration of the disclosed compositions is effective in improving cognition and / or preventing or treating age-related memory loss or cognitive decline. In certain respects, administration of the disclosed composition increases the levels of brain-derived neurotrophic factor (BDNF) and neuroproliferative factor (NGF), or is otherwise neuroprotective.

[0053]

[0065] Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 200 mg per day. Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 300 mg per day. Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 400 mg per day. Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 500 mg per day. Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 750 mg per day. Further details indicate that the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 1000 mg per day. Further, the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 2000 mg per day. Further, the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 3000 mg per day. Further, the disclosed compound increases muscle mass in humans when administered orally at doses greater than approximately 5000 mg per day. [Examples]

[0054]

[0066] The following examples are provided to those skilled in the art to provide a complete disclosure and explanation of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated, and are intended purely as examples of the invention and not to limit the scope of what the inventors consider to be their invention. However, those skilled in the art will recognize, in view of this disclosure, that many variations can be made in particular embodiments and that similar or analogous results can still be obtained without departing from the spirit and scope of the invention.

[0055]

[0067] A case study was conducted on a healthy 24-year-old subject weighing 180 pounds. The subject visited the laboratory in a fasted state prior to the test day. The subject was a resistance-trained individual who performed weight training four days a week. On day 1, the subject was randomized to consume one of two conditions. Following the consumption of a fluid mixture (8 ounces, water), plasma amino acids were measured at 30, 60, 90, and 120 minutes. After a 72-hour rinse, the second condition was performed following the same procedure.

[0056]

[0068] The dosage used in this study was either 2 grams of dileucine (condition Ce) or 2 grams of leucine (condition C4). Leucine

[0069] Table 1. Plasma leucine concentrations (nmOL / L) after ingestion under C4 and CE conditions.

[0057] [Table 1]

[0058]

[0070] Results: As shown in Figure 2, both conditions showed an increase from baseline levels at all time points examined. The relative increase from pre-intake to 30 minutes post-intake was 2.2x higher in condition Ce than in condition C4 (238.07 vs. 106.09 nmol / ml). The relative increase from pre-intake to 60 minutes post-intake was 1.7x higher in condition Ce than in condition C4 (92.81 vs. 54.45 nmol / ml). Condition C4 showed a higher increase in plasma leucine concentration than condition Ce at 90 and 120 minutes compared to baseline levels; the increases were 2.6x and 1.5x higher, respectively. The highest leucine concentration was observed in condition Ce at 30 minutes post-intake (392.03 nmol / l), which was the largest relative increase from the pre-intake level (238.07 nmol / l).

[0059] Isoleucine

[0071] Table 2. Plasma isoleucine concentrations (nmOL / L) after ingestion under C4 and CE conditions.

[0060] [Table 2]

[0061]

[0072] Results: As shown in Figure 3, both conditions showed lower plasma isoleucine levels at 30, 60, 90, and 120 minutes after ingestion (-37.7% to -11.8%).

[0062] Valine

[0073] Table 3. Plasma valine concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0063] [Table 3]

[0064]

[0074] Results: As shown in Figure 4, both conditions showed a decrease in plasma valine concentration over time (-21.3% to -2.3%). After 120 minutes, condition C4 showed a slight increase compared to the pre-intake value (1.9%, 4.35 nmol / l).

[0065] Total BCAA

[0075] Table 4. Total plasma BCAA concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0066] [Table 4]

[0067]

[0076] Results: Figure 5 shows the total plasma BCAA concentrations after ingestion under conditions C4 and Ce. Both conditions showed higher total plasma BCAA concentrations 30 and 60 minutes after ingestion. The peak increase in plasma BCAAs was observed in condition Ce at 30 minutes (35.7%, 213.45 nmol / l); this increase was 2.9 times higher than that of condition C4. The aforementioned results reflect the response observed in plasma leucine concentration. Only condition C4 maintained higher plasma BCAA concentrations compared to pre-ingestion levels at 90 and 120 minutes.

[0068] threonine

[0077] Table 5. Plasma threonine concentrations (nmOL / L) after ingestion under C4 and CE conditions.

[0069] [Table 5]

[0070]

[0078] Results: Figure 6 shows the plasma threonine concentrations after ingestion under conditions C4 and CE. Opposite responses were observed between the conditions regarding plasma threonine concentration. Condition Ce showed an increase in plasma threonine concentration 30 and 60 minutes after ingestion (+6.6% and +5.7%, respectively), while condition C4 had a decrease in levels (-12.5% ​​and -13.2%). 90 and 120 minutes after ingestion, condition C4 had higher plasma threonine levels compared to pre-ingestion values ​​(4.2% and 4.1%, respectively), while condition Ce had lower levels (-3.2% and -1.9%, respectively).

[0071] Methionine

[0079] Table 6. Plasma methionine concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0072] [Table 6]

[0073]

[0080] Results: Figure 7 shows plasma methionine concentrations after ingestion under conditions C4 and CE. Regarding plasma threonine concentration, opposite responses were observed between the conditions. Condition Ce showed an increase in plasma threonine concentration 30 and 60 minutes after ingestion (10.1% and 4.4%, respectively), while condition C4 had a decrease in levels (-11.5% and -15.1%). 90 and 120 minutes after ingestion, condition C4 had higher plasma threonine levels compared to pre-ingestion values ​​(2.8% and 2.3%, respectively), while condition Ce had lower levels (-7.5% and -7.3%, respectively).

[0074] Tryptophan

[0081] Table 7. Plasma tryptophan concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0075] [Table 7]

[0076]

[0082] Results: Figure 8 shows plasma tryptophan concentrations after ingestion under conditions C4 and CE. Both conditions showed a decrease in plasma tryptophan concentration over time (-20.5% to -0.4%). After 90 minutes, condition C4 showed a slight increase from pre-ingestion levels (3.5%, 1.58 nmol / l).

[0077] Phenylalanine

[0083] Table 8. Plasma phenylalanine concentrations (nmOL / L) after ingestion under C4 and CE conditions.

[0078] [Table 8]

[0079]

[0084] Results: Figure 9 shows plasma phenylalanine concentrations after ingestion under conditions C4 and CE. Condition C4 showed lower plasma phenylalanine levels at all time points examined compared to pre-ingestion levels. Alanine concentrations were observed (-16.0% to -3.5%). However, conditional Ce levels were higher at 30 and 60 minutes compared to pre-ingestion levels (6.0% and 3.4%, respectively).

[0080] lysine

[0085] Table 9. Plasma lysine concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0081] [Table 9]

[0082]

[0086] Results: Figure 10 shows plasma lysine concentrations after ingestion under conditions C4 and CE. Both conditions showed an increase in plasma lysine concentration (6.0%–41.3%) 30, 60, and 120 minutes after ingestion. The increases observed at 30 and 60 minutes were higher than 2.8x and 5.7x, respectively, in condition Ce. The increase at 120 minutes was 19x higher in C4 than in condition Ce. Condition Ce experienced a decrease of -18.8% in plasma lysine concentration compared to pre-ingestion levels.

[0083] Total EAA

[0087] Table 10. Total plasma EAA concentrations (nmol / l) after ingestion under C4 and CE conditions.

[0084] [Table 10]

[0085]

[0088] Results: Figure 11 shows the total plasma EAA concentrations after ingestion for conditions C4 and CE. Condition C4 showed increases in total EAA concentrations at 30, 60, and 120 minutes after ingestion compared to baseline (9.4%–16.3%); however, a very small decrease was observed at 60 minutes (-0.6%). Condition Ce showed increases at 30 and 60 minutes (7.2%–26.9%). The greatest increase in total EAA concentration over the time study occurred in condition Ce at 30 minutes (26.9%, 265.57 nmol / l); this increase was 4x higher than in condition C4 at 30 minutes. Finally, condition Ce experienced decreases at 90 and 120 minutes (-11.6% and -6.0%, respectively).

[0086]

[0089] Another exemplary study was conducted on male subjects. These subjects visited the laboratory after an 8-hour fast. Following the 8-hour fast, subjects were provided with a standardized diet plan for two days prior to the study. Resting heart rate, blood pressure, weight, height, and anthropometric measurements using DEXA were performed on the subjects. Furthermore, baseline assessments of clinical safety, amino acid retention, and hydration status were conducted. Therefore, urine samples were collected. In a double-blind manner, venous blood samples (10 mL) were collected from the forearm vein before administering a single dose of either leucine (2 g), dileucine (2 g), or leucine (1 g) + dileucine (1 g) to the subjects. Subsequently, venous blood samples were collected 30, 60, 90, 120, and 240 minutes after ingestion of the assigned composition. After collecting the blood samples, the subjects were given 250 mL of cold water to ingest. A second urine sample was collected 240 minutes after ingestion of the composition. Figure 12 schematically shows the methodology.

[0087]

[0090] All blood samples were processed to enable the determination of amino acid concentrations. Blood samples collected at 0 and 240 minutes were also processed for the determination of a comprehensive metabolic panel and complete blood count (CBC), including platelet differences. Both urine samples were processed to enable the determination of amino acid retention, clinical urine analysis, and basic safety parameters (e.g., creatinine). During processing, all blood and urine samples were stored at -80°C. Subjects returned approximately 3–7 days after the completion of the previous study visit and completed the same study sessions described above while receiving other treatments.

[0088]

[0091] Plasma essential amino acids (threonine, methionine, lysine, histidine, valine, tryptophan, leucine, phenylalanine, isoleucine) were analyzed by LC / MS / MS, and plasma dileucine was analyzed by GC / MS.

[0089]

[0092] Table 11. Leucine-leucine concentration: nmol / mL. Amino acid concentration: μmol / L. BLOD: Leu-Leu concentration in the sample is below detection level.

[0090] [Table 11]

[0091]

[0093] Table 11 shows the individual amino acid concentrations for all three treatments at various time points. The two treatments containing dileucine (2g dileucine and 1g leucine + 1g dileucine) resulted in a measurable increase in plasma dileucine levels. In contrast to leucine (2g) alone, the addition of dileucine (1g) to leucine (1g) delayed the time to reach the maximum leucine concentration. Figures 13 and 14 show the difference between leucine (2g) or leucine. This shows a comparison of leucine Tmax and AUC after administration of 1g of leucine and 1g of dileucine.

[0092]

[0094] In another exemplary study, muscle protein synthesis was measured. Two subjects visited the laboratory after a 7-hour overnight fast. Baseline blood samples were taken. Then, priming (2.0 μmol·kg⁻¹) of L-[cyclic-13C6]-phenylalanine and L-[15N]-phenylalanine, followed by continuous infusion (0.05 μmol·kg⁻¹·min⁻¹) in one catheter was initiated.

[0093]

[0095] The isotope (Cambridge Isotopes Inc., Andover, Massachusetts) was dissolved in 0.9% physiological saline, filtered through a 0.2 μm filter, and injected using a calibrated syringe pump (Harvard Apparatus, Holliston, Massachusetts). After achieving steady-state concentration, L-[15N]-phenylalanine tracer injection was stopped at t=0.

[0094]

[0096] Repeated muscle biopsy samples were taken before and after ingestion of 2 g leucine or 2 g dileucine during an infusion study to determine muscle protein synthesis (based on L-[cyclic-13C6]-phenylalanine uptake). Under anesthesia (lidocaine HCl 1%), biopsies (~100 mg wet weight) were obtained from the central region of the vastus lateralis muscle using a 5 mm Bergstroem needle custom-modified for manual aspiration. A schematic diagram of the experimental protocol can be seen in Figure 15.

[0095]

[0097] Figure 16 shows muscle biopsy-derived muscle protein synthesis in subjects administered either leucine or dileucine. Subjects administered dileucine showed a significantly increased muscle fiber FSR 180 minutes after exposure compared to subjects administered leucine.

[0096]

[0098] While this disclosure has been described in terms of preferred embodiments, those skilled in the art will recognize that variations may be made in form and detail without departing from the spirit and scope of the disclosed apparatus, systems and methods.

Claims

1. A composition for increasing muscle protein synthesis: a. Dileucine; b. Leucine; and c. The pharmaceutically acceptable carrier It contains dileucine in an amount of 30% to 70% (w / w) of the total amount of the composition. The aforementioned composition.

2. The composition according to claim 1, wherein dileucine is present in an amount of 40% to 60% (w / w).

3. The composition according to claim 2, wherein dileucine is present at 50% (w / w).

4. A composition comprising leucine and dileucine for use in a method for treating muscle atrophy in a subject or for preventing muscle atrophy in a subject at risk of muscle atrophy, wherein dileucine is present in an amount of 30% to 70% (w / w) of the total amount of the composition.

5. The composition according to claim 4, wherein dileucine is present in an amount of 40% to 60% (w / w).

6. The composition according to claim 5, wherein dileucine is present at 50% (w / w).

7. The composition according to claim 4, wherein administration of the composition to a subject synergistically increases the plasma level of leucine compared to administration of a composition containing leucine but not containing dileucine.

8. The composition according to claim 4, wherein the subject has sarcopenia.

9. The composition according to claim 8, wherein the composition is administered in a therapeutically effective amount.

10. A composition comprising dileucine, leucine, trileucine, and Leu-Leu-R for use in a method for increasing muscle protein synthesis in a subject, R is an amino acid or amino acid derivative, or a pharmaceutically acceptable salt thereof. Dileucine is present in an amount of 30% to 70% (w / w) of the total amount of the composition, and leucine is present in an amount of 70% to 30% (w / w) of the total amount of the composition. The aforementioned composition.

11. The composition of claim 10, wherein dileucine is present in an amount of 40% to 60% (w / w).

12. The composition of claim 11, wherein dileucine is present at 50% (w / w).

13. The composition according to claim 10, wherein administration of the composition to a subject synergistically increases the plasma level of leucine compared to administration of a composition containing leucine but not containing dileucine.

14. The composition according to claim 10, wherein administration of the composition to a subject synergistically increases muscle mass and / or muscle strength compared to administration of a composition containing leucine but not containing dileucine.

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