Compositions and methods for treating aging-related diseases

Abstract

The present invention relates to compositions and methods for treating aging-related diseases. The present invention provides a composition comprising at least three substances selected from the group consisting of alpha-ketoglutaric acid, malic acid, pterostilbene, micro-dose lithium, glycine, ginger, Rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin, L-theanine, oxaloacetic acid, fumaric acid, succinic acid, hyaluronic acid, butyric acid, anthocyanins, piper longum amide, quercetin, curcumin, and a pharmaceutically acceptable salt thereof. And caffeine, trehalose and spermidine. The present invention also provides related pharmaceutical compositions, methods for improving senescence profiles, and articles of manufacture.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of February 1, 2021, application number 202180011905.7, and invention name “Compositions and methods for treating aging-related diseases”.

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 969,041, filed February 1, 2020, the contents of which are incorporated herein by reference.

[0003] Various publications are cited throughout this application. The disclosures of these publications are incorporated into this application by reference in order to more fully describe the state of the art to which this invention belongs. Technical Field

[0004] The present invention relates to compositions and methods for ameliorating a range of age-related diseases. Background Art

[0005] Aging can be considered as a continuous decline in physical health and an intrinsic loss of organismal function, which results in an increased risk of aging-related diseases, disability, and death.

[0006] Currently, many nutritional and supplement formulations are based on outdated views of aging, such as Denham Harman's Free Radical Theory of Aging (1). This theory posits that aging is primarily caused by free radicals that disrupt and impair cellular homeostasis. According to this outdated theory, antioxidants (e.g., vitamin E, vitamin A, and coenzyme Q10) have the ability to neutralize free radicals, thereby slowing the aging process. However, studies have shown that most antioxidants do not slow aging and may even shorten lifespan or counteract the beneficial effects of exercise (2-6). For these and many other reasons, there is an unmet need for quality oral formulations to improve the hallmarks of aging. Summary of the invention

[0007] The present invention provides a composition comprising at least three substances selected from the following: alpha-ketoglutarate, malate, pterostilbene, micro-dosed lithium, glycine, ginger (preferably ginger extract), Rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin, L-theanine, oxaloacetate, fumarate, succinate, hyaluronic acid, butyrate, anthocyanin, piperlongumine, quercetin, curcuminoids (preferably curcumin), caffeine, trehalose and spermidine.

[0008] The present invention also provides a composition comprising (i) the composition of the present invention and (ii) a pharmaceutically acceptable carrier.

[0009] The present invention also provides a method for improving at least one characteristic of aging in a subject, comprising administering to the subject a composition of the present invention, wherein the characteristic of aging is selected from genomic instability, telomere attrition, epigenetic alteration, loss of proteostasis, deregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

[0010] Finally, the present invention provides an article of manufacture comprising (i) a composition of the present invention and (ii) a nicotinamide-based compound, preferably nicotinamide mononucleotide, in separate compartments. DETAILED DESCRIPTION

[0011] The present invention provides compositions and methods for ameliorating a range of age-related diseases.

[0012] definition

[0013] In this application, certain terms used will have the following meanings.

[0014] In terms of agents, "administering" as used herein means delivering the agent to the subject's body by any known method. Specific modes of administration include, but are not limited to, intravenous, intramuscular, oral, sublingual, transdermal, subcutaneous, intraperitoneal, and intrathecal administration. Preferably, the composition of the present invention is administered orally.

[0015] In addition, in the present invention, one or more conventionally used pharmaceutically acceptable carriers can be used to prepare various compositions. Such carriers are well known to those skilled in the art. For example, oral delivery systems include, for example, tablets, capsules, and powders (e.g., powders that can be added to water to form drinkable preparations). These can include excipients, such as binders (e.g., hydroxypropyl methylcellulose, polyvinyl pyrrolidone, other cellulose materials, and starch), diluents (e.g., lactose and other sugars, starch, dicalcium phosphate, and cellulose materials), disintegrants (e.g., starch polymers and cellulose materials), and lubricants (e.g., stearates and talc). In one embodiment, the oral delivery system is a powder and includes one or more of xylitol, malic acid, flavoring agents, silicon dioxide, calcium silicate, and rebaudioside A.

[0016] As used herein, the term "signatures of aging" (used synonymously with "age-related signatures" and "signatures associated with aging") includes, but is not limited to, any of the following: genomic instability, telomere attrition, epigenetic alterations, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. These signatures are known and have been extensively studied by C. López Otín, et al. (7).

[0017] As used herein, the term "ameliorating" when used in reference to a characteristic of aging includes, but is not limited to (i) slowing, halting, reversing, or preventing the progression of the characteristic, (ii) slowing, halting, reversing, or preventing the progression of a symptom of the characteristic, (iii) preventing or reducing the likelihood of recurrence of the characteristic, and / or (iv) preventing or reducing the likelihood of recurrence of a symptom of the characteristic. In one embodiment, treating a subject having a characteristic of aging means (i) reversing the progression of the characteristic, ideally to the point of eliminating the characteristic, and / or (ii) reversing the progression of a symptom of the characteristic, ideally to the point of eliminating the symptom.

[0018] The term "subject" as used herein includes, but is not limited to, mammals, such as humans, non-human primates, dogs, cats, horses, sheep, goats, cattle, rabbits, pigs, rats, and mice. In the case where the subject is a human, the subject can be of any age. For example, the subject can be 60 years old or older, 65 years old or older, 70 years old or older, 75 years old or older, 80 years old or older, 85 years old or older, or can be 90 years old or older. Alternatively, the subject can be 50 years old or younger, 45 years old or younger, 40 years old or younger, 35 years old or younger, 30 years old or younger, 25 years old or younger, or can be 20 years old or younger.

[0019] Embodiments of the invention

[0020] The present invention provides compositions and methods for improving a range of age-related diseases. These compositions contain certain combinations of substances that act synergistically to address aging characteristics such as genomic instability, telomere attrition, epigenetic changes, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. The present invention also provides related products.

[0021] In particular, the present invention provides a composition comprising at least three substances selected from the following: alpha-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin, L-theanine, oxaloacetic acid, fumaric acid, succinic acid, hyaluronic acid, butyric acid, anthocyanins, piperamide, quercetin, curcuminoids (preferably curcumin), caffeine, trehalose and spermidine. In one embodiment, the composition comprises at least three substances selected from the following: alpha-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger extract, rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin and L-theanine.

[0022] Each of these substances is commercially available in oral dosage forms. The following are some non-limiting examples. Alpha-ketoglutarate, also known as alpha-ketoglutaric acid, is commercially available, for example, in 1,000 mg capsules. Alpha-ketoglutaric acid is available as a calcium salt, but all other salts of alpha-ketoglutaric acid are contemplated in the present invention. Malic acid, also known as malic acid, is commercially available, for example, as magnesium malate in 500 mg capsules. Also contemplated as alternatives to malic acid for use in the present invention are fumaric acid and oxaloacetic acid. Pterostilbene is commercially available, for example, in 100 mg capsules. Microdose lithium, also known as trace lithium, is commercially available, for example, in 1,000 mcg capsules. Glycine is commercially available, for example, in 1,000 mg capsules. Ginger as ginger root and ginger extract (preferably organic ginger extract, standardized to 2% gingerols, such as PurGinger TM (Applied Food Sciences) is commercially available. Rhodiola rosea, preferably with a minimum of 3% rosavin and 2% salidroside, is commercially available, for example, in 300 mg capsules. Acetyl glucosamine, also known as N-acetylglucosamine, is commercially available, for example, in 600 mg capsules. Glucosamine can be used alone in its salt form (e.g., glucosamine sulfate, HCl glucosamine, 2KCl glucosamine) and in its acylated form (i.e., acetylglucosamine). Acetyl glucosamine is also a component of hyaluronic acid, an acetylglucosamine-containing polysaccharide. Therefore, acetylglucosamine can also be delivered to a subject in the form of hyaluronic acid. Vitamin C, also known as ascorbic acid and ascorbate, is commercially available, for example, in 1,000 mg tablets. Glucosamine is commercially available, for example, as glucosamine sulfate, in capsules containing 400 mg glucosamine sulfate. Fisetin is commercially available, for example, in 100 mg capsules. L-Theanine, also known as L-γ-ethylglutamine and N 5 -ethyl-L-glutamine is commercially available, for example, in 100 mg capsules. Spermidine is commercially available, for example, in 400 mg capsules of wheat germ extract containing 0.5 mg spermidine. Finally, trehalose is commercially available in powder form.

[0023] Various combinations of substances are contemplated for the compositions of the present invention. In one embodiment, the compositions of the present invention comprise alpha-ketoglutarate, microdose lithium, and fisetin. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, fisetin, pterostilbene, and glycine. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, microdose lithium, and pterostilbene. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, microdose lithium, and glycine. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, pterostilbene, microdose lithium, and fisetin. In another embodiment, the compositions of the present invention comprise pterostilbene, microdose lithium, and fisetin. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, microdose lithium, fisetin, glycine, and pterostilbene. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, microdose lithium, fisetin, glycine, and vitamin C. In another embodiment, the compositions of the present invention comprise alpha-ketoglutarate, pterostilbene, microdose lithium, glycine, glucosamine, and fisetin. Additional embodiments are set forth in the Examples section below.

[0024] In another embodiment, the composition of the present invention comprises alpha-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger (preferably ginger extract), rhodiola rosea, hyaluronic acid, vitamin C, glucosamine, fisetin and L-theanine. Preferably, the composition of the present invention comprises 1,100 mg calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin and 150 mg L-theanine. In another embodiment, the composition of the present invention comprises 1,100 mg calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin, 200 mg L-theanine, xylitol, malic acid, flavoring agents, silicon dioxide, calcium silicate, and rebaudioside A. Optionally, the composition of the present invention comprises one or more sweeteners, such as, but not limited to, erythritol, trehalose, maltitol, mannitol, sorbitol, xylitol, isomalt, lactitol, tagatose, monk fruit, inuline, and stevia-derived substances. Preferably, the compositions of the invention (eg, the embodiments described in this paragraph) are in the form of a powder that, when mixed with a liquid such as water, can be ingested orally (preferably once daily) by a subject as a beverage.

[0025] Preferably, one or more substances in the composition of the present invention are formulated for delayed release and / or extended release. In one embodiment of the composition of the present invention, alpha-ketoglutarate, microdose lithium, fisetin, butyric acid, pterostilbene and glycine are formulated for delayed release and / or extended release. In another embodiment of the composition of the present invention, each substance is formulated for delayed release and / or extended release. In another embodiment, the composition of the present invention comprises 1,100 mg delayed release and / or extended release calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg delayed release and / or extended release pterostilbene, 1 mg delayed release and / or extended release microdose lithium, 2,000 mg delayed release and / or extended release glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg delayed release and / or extended release fisetin, 150 mg L-theanine, xylitol, malic acid, flavoring agents, silicon dioxide, calcium silicate, and rebaudioside A. Optionally, the composition of the present invention comprises one or more sweeteners, such as, but not limited to, erythritol, trehalose, maltitol, mannitol, sorbitol, xylitol, isomalt, lactitol, tagatose, monk fruit, inulin, or a stevia-derived substance.

[0026] The present invention also provides a composition comprising (i) the composition of the present invention and (ii) a pharmaceutically acceptable carrier.

[0027] The present invention also provides a method for improving at least one aging feature in a subject, comprising administering a composition of the present invention to a subject (preferably once a day), wherein the aging feature is selected from genomic instability, telomere loss, epigenetic changes, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. In one embodiment, the method of the present invention improves genomic instability. In another embodiment, the method of the present invention improves telomere loss. In another embodiment, the method of the present invention improves one or more epigenetic changes. In another embodiment, the method of the present invention improves loss of protein homeostasis. In another embodiment, the method of the present invention improves dysregulated nutrient sensing. In another embodiment, the method of the present invention improves mitochondrial dysfunction. In another embodiment, the method of the present invention improves cellular senescence. In another embodiment, the method of the present invention improves stem cell exhaustion. In another embodiment, the method of the present invention improves altered intercellular communication.

[0028] In another embodiment of the methods of the present invention, the methods improve at least two characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least three characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least four characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least five characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least six characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least seven characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least eight characteristics of aging in a subject. In another embodiment, the methods of the present invention improve at least nine characteristics of aging in a subject.

[0029] For example, in one embodiment, the method of the present invention improves at least the following aging characteristics in a subject: genomic instability, epigenetic changes, and loss of protein homeostasis. In another embodiment, the method of the present invention improves at least the following aging characteristics in a subject: epigenetic changes, loss of protein homeostasis, and mitochondrial dysfunction. In another embodiment, the method of the present invention improves at least the following aging characteristics in a subject: epigenetic changes, loss of protein homeostasis, and dysregulated nutrient sensing. In another embodiment, the method of the present invention improves cellular senescence or reduces the amount of senescent cells. In another embodiment, the method of the present invention improves cellular senescence and stem cell exhaustion. In another embodiment, the method of the present invention improves mitochondrial dysfunction and loss of protein homeostasis. In another embodiment, the method of the present invention improves epigenetic changes and loss of protein homeostasis. In another embodiment, the method of the present invention improves intercellular communication changes and dysregulated nutrient sensing. In another embodiment, the method of the present invention improves intercellular communication changes and stem cell exhaustion. In another embodiment, the method of the present invention improves stem cell exhaustion and epigenetic changes. In another embodiment, the method of the present invention improves epigenetic changes and mitochondrial dysfunction. In another embodiment, the method of the present invention improves epigenetic changes, loss of protein homeostasis, and mitochondrial dysfunction. In another embodiment, the methods of the present invention improve stem cell exhaustion, cell senescence, and altered intercellular communication. In another embodiment, the methods of the present invention improve epigenetic changes, cell senescence, and mitochondrial dysfunction. In another embodiment, the methods of the present invention improve nutrient sensing disorders, mitochondrial dysfunction, and loss of protein homeostasis. In another embodiment, the methods of the present invention improve epigenetic changes, loss of protein homeostasis, mitochondrial dysfunction, and cell senescence.

[0030] Preferably, the method of the present invention further comprises administering to the subject a nicotinamide-based compound (preferably nicotinamide mononucleotide) simultaneously. In the present invention, "simultaneous administration" of a nicotinamide-based compound and the composition of the present invention includes, for example, (i) administering the nicotinamide-based compound at the same time as the composition of the present invention, or (ii) administering the nicotinamide-based compound within one hour, three hours, six hours, twelve hours, or twenty-four hours of the composition of the present invention. Nicotinamide-based compounds include, but are not limited to, nicotinamide adenine dinucleotide (NAD), nicotinamide mononucleotide, nicotinic acid (i.e., nicotinic acid), nicotinamide (NAM), nicotinamide riboside (NR), nicotinic acid riboside (NaR), and nicotinic acid mononucleotide (NaNM). In a preferred embodiment, the method of the present invention further comprises administering to the subject 125 mg of nicotinamide mononucleotide in capsule form simultaneously. Nicotinamide mononucleotide (also known as NMN, nicotinamide ribonucleoside 5'-phosphate, nicotinamide D-ribonucleotide, β-nicotinamide ribose monophosphate, and nicotinamide nucleotide) is commercially available in capsule form at various doses (eg, 100 mg and 300 mg).

[0031] In the present invention, the methods of the invention are expected to ameliorate genomic instability, telomere attrition, one or more epigenetic changes, loss of proteostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

[0032] Preferably, in the methods of the present invention, the subject is a human. In another embodiment, the subject is a cat, dog or horse.

[0033] Finally, the present invention provides such an article, which comprises (i) a composition of the present invention and (ii) a nicotinamide-based compound (preferably nicotinamide mononucleotide) in separate compartments. In one embodiment, the article of the present invention comprises (i) a composition comprising alpha-ketoglutarate, malic acid, pterostilbene, microdose lithium, glycine, ginger (preferably ginger extract), rhodiola rosea, hyaluronic acid, vitamin C, glucosamine, fisetin and L-theanine, and (ii) a nicotinamide-based compound (preferably nicotinamide mononucleotide) in separate compartments.

[0034] Preferably, the article of manufacture of the present invention comprises in separate compartments (i) a composition containing 1,100 mg calcium α-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin and 150 mg L-theanine (and optionally also containing xylitol, malic acid, flavoring agents, silicon dioxide, calcium silicate and rebaudioside A) (preferably in powder form), and (ii) 125 mg nicotinamide mononucleotide (preferably in capsule form).

[0035] In the methods of the present invention, it is contemplated that the subject is orally administered a composition of the present invention daily, preferably simultaneously with a nicotinamide-based compound (preferably nicotinamide mononucleotide). The following doses are contemplated, but are not limited thereto: (i) 1,100 mg calcium α-ketoglutarate (or optionally, any of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 1,200 mg, 1,300 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,700 mg, 1,800 mg, 1,900 mg, or 2,000 mg); (ii) 2,000 mg magnesium malate (or optionally, any of 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg , 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg, 2,200 mg, 2,400 mg, 2,600 mg, 2,800 mg, 3,000 mg, 3,200 mg, 3,400 mg, 3,600 mg, 3,800 mg, or 4,000 mg); (iii) 50 mg pterostilbene (or optionally, any of 10 mg, 20 mg, 30 mg, 40 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg); (iv) 1 mg microdose lithium (or optionally, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg , 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg or 2.0 mg); (v) 2,000 mg glycine (or optionally, 200 mg, 400 mg, 600 mg, 800 mg, 1,000 mg, 1,200 mg, 1,400 mg, 1,600 mg, 1,800 mg, 2,200 mg, 2,400 mg, 2,600 mg, 2,800 mg, 3,000 mg, 3,200 mg, 3 , 400 mg, 3,600 mg, 3,800 mg or 4,000 mg); (vi) 80 mg ginger extract (or optionally, any of 10 mg, 20 mg, 30 mg, 40 mg, 60 mg, 70 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg or 1,000 mg);(vii) 300 mg of Rhodiola rosea (or optionally, any of 100 mg, 150 mg, 200 mg, 250 mg, 350 mg, 400 mg, 450 mg or 500 mg); (viii) 100 mg of hyaluronic acid (or optionally, any of 10 mg, 20 mg, 30 mg, 40 mg, 60 mg, 70 mg, 80 mg, 90 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg); g, 180 mg, 190 mg, or 200 mg); (ix) 100 mg vitamin C (or optionally, any of 10 mg, 20 mg, 30 mg, 40 mg, 60 mg, 70 mg, 80 mg, 90 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, or 200 mg); (x) 1,000 mg glucosamine sulfate (or optionally, any of 200 (xi) 100 mg of fisetin (or optionally, 10 mg, 20 mg, 30 mg, 40 mg, 60 mg, 70 mg, 80 mg, 90 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 1 90 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 1,100 mg, 1,200 mg, 1,300 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,700 mg, 1,800 mg, 1,900 mg or 2,000 mg); (xii) 150 mg L-theanine (or optionally, any one of 20 mg, 40 mg, 60 mg, 80 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240 mg, 260 mg, 280 mg or 300 mg); (xiii) 3 mg spermidine (or optionally, any one of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg or 12 mg); (xiv) 15 g trehalose (or optionally, any one of 1 g, 2 g, 5 g, 10 g, 20 g, 30 g, 40 g or 50 g);and (xv) 125 mg nicotinamide mononucleotide (or optionally, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, any one of 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1,000 mg, 1,100 mg, 1,200 mg, 1,300 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,700 mg, 1,800 mg, 1,900 mg or 2,000 mg). ;

[0036] The following three dosage regimens (i) are also intended for daily oral administration (in delayed release and / or extended release form, as well as in non-delayed release, non-extended release form) and (ii) are intended for use in the compositions and articles of manufacture of the present invention. They include, but are not limited to, the following: (i) 275 mg calcium α-ketoglutarate, 500 mg magnesium malate, 12.5 mg pterostilbene, 0.25 mg microdose lithium, 500 mg glycine, 20 mg ginger extract, 75 mg rhodiola rosea, 25 mg hyaluronic acid, 25 mg vitamin C, 250 mg glucosamine sulfate, 25 mg fisetin, and 37.5 mg L-theanine; (ii) 550 mg calcium α-ketoglutarate, 1,000 mg magnesium malate, 25 mg pterostilbene, 0.5 mg microdose lithium, 1,000 mg glycine, 40 mg ginger extract, 150 mg rhodiola rosea, 50 mg hyaluronic acid, 50 mg vitamin C, 500 mg glucosamine sulfate, 50 mg fisetin, and 75 mg L-theanine; and (iii) 825 mg calcium alpha-ketoglutarate, 1,500 mg magnesium malate, 37.5 mg pterostilbene, 0.75 mg microdose lithium, 1,500 mg glycine, 60 mg ginger extract, 225 mg rhodiola rosea, 75 mg hyaluronic acid, 75 mg vitamin C, 750 mg glucosamine sulfate, 75 mg fisetin, and 112.5 mg L-theanine.

[0037] The present invention will be better understood by reference to the following examples, however, those skilled in the art will readily appreciate that the specific examples detailed are merely illustrative of the invention as more fully described in the claims which follow.

[0038] Example

[0039] Example 1 - Other embodiments of the compositions of the present invention

[0040] For the compositions of the present invention, combinations of the following additional substances are contemplated but not limited.

[0041] The first additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin and one or more additional substances selected from the group consisting of malic acid, pterostilbene, glycine, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine and L-theanine.

[0042] The second additional combination comprises alpha-ketoglutarate, microdose lithium, pterostilbene and one or more additional substances selected from the group consisting of malic acid, glycine, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin and L-theanine.

[0043] A third additional combination comprises alpha-ketoglutarate, microdose lithium, glycine and one or more additional substances selected from the group consisting of malic acid, pterostilbene, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin and L-theanine.

[0044] A fourth additional combination comprises alpha-ketoglutaric acid, pterostilbene, fisetin and one or more additional substances selected from the group consisting of malic acid, microdose lithium, glycine, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine and L-theanine.

[0045] A fifth additional combination comprises pterostilbene, microdose lithium, fisetin, and one or more additional substances selected from the group consisting of alpha-ketoglutaric acid, malic acid, glycine, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, and L-theanine.

[0046] A sixth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, glycine, vitamin C, and one or more additional substances selected from the group consisting of malic acid, pterostilbene, ginger (preferably ginger extract), rhodiola rosea, acetylglucosamine, glucosamine, and L-theanine.

[0047] A seventh additional combination comprises alpha-ketoglutaric acid, pterostilbene, microdose lithium, glycine, glucosamine, fisetin, and one or more additional substances selected from the group consisting of malic acid, ginger (preferably ginger extract), rhodiola rosea, vitamin C, glucosamine, and L-theanine.

[0048] An eighth additional combination comprises glucosamine, malic acid, glycine, lithium, pterostilbene, fisetin, hyaluronic acid, alpha-ketoglutaric acid, ginger (preferably ginger extract), anthocyanin, rhodiola, spermidine, vitamin C, or a combination thereof.

[0049] A ninth additional combination comprises glucosamine, malic acid, glycine, lithium, pterostilbene, fisetin, alpha-ketoglutaric acid, ginger (preferably ginger extract), anthocyanin, rhodiola rosea, spermidine, vitamin C, or a combination thereof.

[0050] A tenth additional combination comprises glucosamine, malic acid, glycine, lithium, pterostilbene, fisetin, alpha-ketoglutaric acid, ginger (preferably ginger extract), anthocyanin, spermidine, vitamin C, or a combination thereof.

[0051] An eleventh additional combination comprises glucosamine, malic acid, glycine, lithium, pterostilbene, fisetin, alpha-ketoglutarate, anthocyanin, spermidine, vitamin C, or a combination thereof.

[0052] A twelfth additional combination comprises glucosamine, malic acid, glycine, lithium, pterostilbene, fisetin, alpha-ketoglutaric acid, anthocyanin, vitamin C, or a combination thereof.

[0053] A thirteenth additional combination comprises lithium, malic acid, and alpha-ketoglutarate, with or without vitamin C.

[0054] A fourteenth additional combination comprises lithium, glycine, malic acid, and alpha-ketoglutarate, with or without vitamin C.

[0055] A fifteenth additional combination comprises lithium, glycine, malic acid, fisetin, and alpha-ketoglutarate, with or without vitamin C.

[0056] A sixteenth additional combination comprises alpha-ketoglutarate, microdose lithium, and glucosamine.

[0057] A seventeenth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and pterostilbene.

[0058] An eighteenth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and glucosamine.

[0059] A nineteenth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and glycine.

[0060] A twenty additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, pterostilbene, and glucosamine.

[0061] A twenty-first additional combination comprises alpha-ketoglutarate, fisetin, and spermidine.

[0062] A twenty-second additional combination comprises alpha-ketoglutarate, fisetin, spermidine, and trehalose.

[0063] A twenty-third additional combination comprises alpha-ketoglutarate, fisetin, spermidine, and pterostilbene.

[0064] A twenty-fourth additional combination comprises alpha-ketoglutarate, fisetin, and trehalose.

[0065] A twenty-fifth additional combination comprises alpha-ketoglutarate, fisetin, trehalose, and pterostilbene.

[0066] A twenty-sixth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and spermidine.

[0067] A twenty-seventh additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and trehalose.

[0068] A twenty-eighth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, spermidine, and trehalose.

[0069] A twenty-ninth additional combination comprises alpha-ketoglutarate, fisetin, and butyric acid.

[0070] A thirty additional combination comprises alpha-ketoglutarate, fisetin, pterostilbene, and butyric acid.

[0071] A thirty-first additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and butyric acid.

[0072] A thirty-second additional combination comprises malic acid, glycine, lithium, pterostilbene, fisetin, alpha-ketoglutaric acid, anthocyanin, vitamin C, or a combination thereof.

[0073] A thirty-third additional combination comprises fisetin, pterostilbene, and glucosamine.

[0074] A thirty-fourth additional combination comprises fisetin, glycine, and pterostilbene.

[0075] A thirty-fifth additional combination comprises fisetin, glucosamine, and glycine.

[0076] A thirty-sixth additional combination comprises pterostilbene, glycine, and glucosamine.

[0077] A thirty-seventh additional combination comprises fisetin, pterostilbene, glucosamine, and glycine.

[0078] A thirty-eighth additional combination comprises fisetin, pterostilbene, glucosamine, glycine, and malic acid.

[0079] A thirty-ninth additional combination comprises fisetin, pterostilbene, and acetylglucosamine.

[0080] A forty additional combination comprises fisetin, pterostilbene, glucosamine, glycine and acetylglucosamine.

[0081] A forty-first additional combination comprises fisetin, pterostilbene, and theanine.

[0082] A forty-second additional combination comprises alpha-ketoglutarate, fisetin, and theanine.

[0083] A forty-third additional combination comprises alpha-ketoglutarate, microdose lithium, and theanine.

[0084] A forty-fourth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, and theanine.

[0085] A forty-fifth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, pterostilbene, and theanine.

[0086] A forty-sixth additional combination comprises alpha-ketoglutarate, microdose lithium, fisetin, pterostilbene, glucosamine, and theanine.

[0087] For each of the above additional combinations, it is contemplated that a nicotinamide-based compound (preferably NMN) will be simultaneously administered to the subject.

[0088] For the compositions of the present invention, the following combinations of substances comprising NMN are also contemplated but not limited to: (i) a composition comprising nicotinamide mononucleotide, α-ketoglutarate and fisetin; (ii) a composition comprising nicotinamide mononucleotide, α-ketoglutarate, microdose lithium and fisetin; (iii) a composition comprising nicotinamide mononucleotide, α-ketoglutarate, pterostilbene and fisetin; (iv) a composition comprising nicotinamide mononucleotide, pterostilbene and fisetin; (v) a composition comprising nicotinamide mononucleotide, pterostilbene, fisetin and microdose lithium; and (vi) a composition comprising nicotinamide mononucleotide, pterostilbene, fisetin and glucosamine.

[0089] Example 2 - Measuring epigenetic dysregulation

[0090] The success of the compositions of the invention in ameliorating epigenetic dysregulation in a subject (e.g., a human subject) can be measured, for example, by making any of the following determinations: (i) the subject's epigenetic methylation, histone modification and / or chromatin pattern more closely resembles that of a younger person (e.g., at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% similar); (ii) the epigenetic clock is increased in magnitude, or more closely resembles that of a younger person; (iii) an epigenetic DNA methylation clock (e.g., a Horvath clock or a Levine clock) that measures methylation of DNA in different regions of a subject's DNA that more closely resembles the DNA methylation pattern of a younger subject (e.g., at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% similar); (iv) an epigenetic DNA methylation clock (e.g., a Horvath clock or a Levine clock) that shows that an adult subject has been rejuvenated by at least one year, at least two years, or at least three years following administration of a composition of the invention (or wherein the epigenetic age or biological age has been reduced by at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70% compared to the subject's chronological age); , at least 80%, at least 90%, or at least 100% similar); (iv) the subject's chromatin profile more closely resembles the chromatin profile of a younger subject (e.g., at least 0.2%, at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% similar); (v) the subject's transcriptome more closely resembles the transcriptome of a younger subject. (vi) the expression profile of ribosome-associated proteins and / or ribosome-associated RNA in the subject more closely resembles the expression profile of a younger organism (e.g., at least 0.2%, at least 0.5%, at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% similar).or (vii) the subject's sirtuin levels have been increased (e.g., increased by at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0091] The success of the compositions of the invention in ameliorating epigenetic disorders in a subject (e.g., a human subject) can also be measured, for example, by performing any of the following determinations: (i) an increase in the level of H3K9me3 in a subject (e.g., an increase of at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); or (ii) an increase in the level of a heterochromatin protein (Hp1y) in a subject (e.g., an increase of at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0092] Example 3 - Measuring Cellular Senescence

[0093] The success of the compositions of the invention in ameliorating cellular senescence in a subject (e.g., a human subject) can be measured, for example, by performing any of the following determinations: (i) a delay in the appearance of senescent cells or a delay in the transition of normal cells to senescent cells in vitro or in vivo (e.g., a delay of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); and / or (ii) a reduction in the number of senescent cells in vitro or in vivo (e.g., a reduction of at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). Senescence or a senescence-like state of a cell can be measured, for example, by one or more of the following: (i) an epigenetic profile, such as measuring epigenetic markers of senescence (e.g., changes in histone H3 lysine 9 and 27 trimethylation (altered levels of H3K9me3 and H3K27me3), altered levels of heterochromatin protein 1 (HP1) family proteins, increased levels of the histone variant macroH2A, and / or levels of the chromatin remodeling enzyme ATRX); (ii) changes in cell morphology that are similar to the morphology of senescent cells (e.g., increased size, flatter shape, accumulation of polyploid nuclei or DNA damage foci); (iii) changes in the levels of nuclear lamina propria-associated proteins (e.g., lamins) (e.g., decreased lamin B1); (iv) changes in heterochromatin that are more similar to the heterochromatin state of senescent cells (e.g., increased senescence-associated heterochromatic foci (SAHF)); (v) senescence-associated distention of (vi) changes in the secretory phenotype corresponding to senescent cells (e.g., the presence of a greater number of senescence-associated secretomes, which include but are not limited to cytokines, interleukins (e.g., IL-1, IL-2, IL-6, IL-8, and TNF-α), matrix metalloproteinases, pro-angiogenic factors, pro-inflammatory substances, and growth factors (e.g., vascular endothelial growth factor, insulin-like growth factor, and GM-CSF)).

[0094] The success of the compositions of the invention in improving cellular senescence in a subject (e.g., a human subject) can also be measured, for example, by performing any of the following determinations: (i) an increase in the level of H3K9me3 in a subject (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (ii) an increase in the level of H3K27me3 in a subject (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (iii) a decrease in the level of P16INK4A in a subject (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); or (iv) a decrease in the level of one or more of p16, p53, p21, p14, p15, and β-galactosidase activity in a subject (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0095] Example 4 - Measuring changes in intercellular communication

[0096] The success of the compositions of the invention in improving alterations in intercellular communication in a subject (e.g., a human subject) can be measured, for example, by determining a decrease (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%) in the level of one or more of IL-1, IL-2, IL-6, IL-8, IL-17a, and TNF-α in the subject.

[0097] Example 5 - Measuring Autophagy

[0098] The success of the compositions of the invention in improving reduced autophagy in a subject (e.g., a human subject) can be measured, for example, by determining any of the following: (i) an increase in the level of one or more of LC3, p62, Ulk1, Parkin, PINK1, LAMP2A, Atg, FIP200, Vps15, Beclin, Lamp-1, Lamp-2, Hsp70, Hsp90, and SQSTM1 in the subject (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (ii) an increase in any of phagophore formation, autophagosome formation, lysosome number, lysosomal activity, LC3 spots, lysosomal content (as determined, for example, using immunochemistry, electron microscopy, Western blotting, or flow cytometry) (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). (iii) an increase in lysosomal acidity (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (iv) upregulation of the ubiquitin-proteasome system or (v) inhibition of mTOR or the mTOR pathway (e.g., inhibition of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0099] Example 6 - Measuring Cellular Oxidation

[0100] The success of the compositions of the invention in improving cellular oxidation in a subject (e.g., a human subject) can be measured, for example, by performing any of the following determinations: (i) a decrease in the level of 8-isoprostane in a subject (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (ii) a decrease in the level of 8-hydroxydeoxyguanosine (8-OHdG) in a subject (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); or (iii) a decrease in the level of oxidized LDL, malondialdehyde (MDA), 4-HNA, lipid hydroperoxides, oxidized proteins, protein carbonylation and protein nitration (3-nitrotyrosine), advanced glycation end products (AGEs) in a subject.

[00136] In some embodiments, the present invention provides a reduction in the level of any of EGFR (EGFR) products, AGEs, and advanced oxidation protein products (e.g., a reduction of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0101] Example 7 - Measuring Telomere Fitness

[0102] The success of the compositions of the invention in ameliorating telomere attrition in a subject (e.g., a human subject) can be measured, for example, by performing any of the following determinations: (i) an increase in telomere length in a subject (e.g., an increase of at least 0.01%, at least 0.1%, at least 0.5%, at least 1.0%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%); or (ii) an increase in mRNA expression of telomerase, TERT, TERC, or a telomerase-related gene (e.g., an increase of at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0103] Example 8 - Measuring stem cell fitness

[0104] The success of the compositions of the invention in improving stem cell exhaustion in a subject (e.g., a human subject) can be measured, for example, by determining an increase in the number of stem cells in vivo (e.g., an increase of at least 1%, at least 2%, at least 5%). This increase can be measured in vitro or in vivo by, for example, one or more of the following biomarkers: (i) muscle satellite stem cell biomarkers (e.g., PAX7, LMNA, and emerin); (ii) neuronal stem cell biomarkers (e.g., nestin, Sox2, ASCL1 / Mash1, ABCG2, BMI-1, β-catenin, Brg1, N-cadherin, calcitonin R, CD15 / Lewis X, CD133, CDCP1, COUP-TF I / NR2F1, CXCR4, FABP7 / B-FABP, FABP8 / M-FABP, FGFR2, FGFR4, FoxD3, Frizzled receptor-9, GCNF / NR6A1, GFAP, Glut1, HOXB1, ID2, LRTM1, Meteorin, MSX1, Musashi-1, Musashi-2, Nestin, NeuroD1, Noggin, Notch-1, Notch-2, Nrf2, Nuclear stem cell factor (Nucleostemin), Numb, Otx2, Pax3, Pax6, PDGF Rα, PKCζ, Prominin 2, ROR2, RUNX1 / CBFA2, RXRα / NR2B1 (e.g., sFRP-2, SLAIN1, SOX1, SOX2, SOX9, SOX11, SOX21, SSEA-1, SSEA-4, TRAF-4, vimentin, and ZIC1); (iii) pluripotent stem cell markers (e.g., Oct4, NANOG, Sox2, and Myc); (iv) hematopoietic stem cell markers (e.g., CD34, CD59, and CD90 / Thy1); and (v) mesenchymal stem cell markers (e.g., CD105, CD90, CD73, CD44, CD45, CD29, CD166, Stro-1, CD106, and GSTT1).

[0105] Because measuring stem cells in vivo is difficult and expensive, the success of the compositions of the invention in ameliorating stem cell depletion can also be measured, for example, by performing the following preferred in vitro ("lab dish") assays (as well as in vivo assays): (i) an increase in stem cell number (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (ii) survival of the stem cells; (iii) an increase in the potency of the stem cells in vitro (and in vivo) (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (iv) an increase in the potency of the stem cells in vitro (and in vivo) (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). (iv) an increase in stem cell quality as measured, for example, by an increase in stem cell growth, an increase in proliferation capacity, an increase in the ability to form colonies, or an increase in the ability to generate cells (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (v) an increase in stem cell lifespan as measured in hours (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). or (vi) an increase (e.g., an increase of at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%) in the resistance of stem cells to a cellular stressor (e.g., heat, cold, or a toxin).

[0106] The success of the compositions of the invention in ameliorating stem cell exhaustion in a subject (e.g., a human subject) can also be measured, for example, by any of the following determinations in vitro and in vivo: (i) as measured, for example, by replication capacity, potency, proliferation capacity, survival (as measured, for example, over time (e.g., the number of additional survival times) by exposing stem cells to physiological stressors and toxins), efficacy, and quality (e.g., the stem cells have an increased number of replications or can give rise to a greater number of cells derived from the stem cells or show increased cell division markers (e.g., cyclin D1 or (i) an improvement in stem cell function as measured by increased B-catechins (e.g., an improvement of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (ii) an improvement in the stem cell transcriptome (i.e., the transcriptome of the stem cells is more similar to the transcriptome of younger stem cells or more functional stem cells) (e.g., an improvement of at least 0.5%, at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%). , at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); (iii) an improvement in the stem cell proteome (i.e., the proteome of the stem cells is more similar to the proteome of younger stem cells or more functional stem cells) (e.g., an improvement of at least 0.5%, at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); or ( iv) an improvement in the stem cell epigenome (i.e., the epigenome of the stem cells is more similar to the epigenome of younger stem cells or more functional stem cells) as assessed, for example, by measuring methylation or histone modification patterns and comparing them to younger, more functional, or more potent stem cells (e.g., an improvement of at least 0.5%, at least 1%, at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0107] Example 9 - Measurement of physiological biomarkers as indicators of stem cell fitness and senescent cell levels in an organism Markers

[0108] The success of the compositions of the invention in improving an adverse physiological event in a subject (e.g., a human subject) can be measured, for example, by making any of the following determinations: (i) a reduction in the number and / or severity of skin wrinkles, as measured, for example, by wrinkle surface area (e.g., by 3D topography or profilometry); (ii) an increase in skin radiance, as measured, for example, by an identometer, a cutometer, a transepidermal water loss meter, a corneometer, or a skin transcriptome or proteome biomarker panel, which indicates improved skin function; (iii) an improvement in cognition, as measured, for example, by cognitive testing; (iv) an improvement in well-being, as measured, for example, by a depressive symptom scale; (v) an improvement in activity, as measured, for example, by an international frailty scale or an activity scale (e.g., the Canadian Study on Health & Aging Clinical Frailty Scale). (vi) improvements in reaction time; (vii) improvements in neuromuscular capacity and strength (e.g., improved endurance, improved balance, improved proprioception, improved mobility (as measured by the 6-minute walk test (6MWT), 4-m gaitspeed test (4MGST), improved short physical performance battery (SPPB) score, or tests measuring balance, gait speed test, chair stand test, or other markers of physical function), improved grip strength, improved power, increased activity level, improved 400-m walk test, and improved arm flexion test results); (viii) improvements in strength and function (e.g., as measured by 100% stamina per mm 3 The invention relates to the present invention to a method for the treatment of a chronic obstructive pulmonary disease (COPD) that is characterized by improved blood flow, as measured by an increase in blood units to tissues; (ix) a decrease in blood pressure; (x) an improvement in vascular health, as measured, for example, by pulse wave velocity or flow-mediated vasodilation of the blood vessels; (xi) resulting in weight loss, a decrease in waist circumference or BMI, or a decrease in abdominal fat volume; (xii) an improvement in heart rate variability (HRV); and (xiii) an improvement in AI-assessed facial aging curves (e.g., as measured by facial AI software that assesses, for example, wrinkle area, blood flow, or skin sagging).

[0109] Importantly, if stem cell health improves and cellular senescence decreases, many of the physiological biomarkers discussed in this example will also improve. This is particularly true for stem cells, as reducing senescence markers of stem cell decline (measurements of which are discussed above) improves physiological biomarkers.

[0110] Example 10 - Measuring genomic fitness

[0111] The success of the compositions of the invention in ameliorating genomic instability in a subject (e.g., a human subject) can be measured, for example, by determining a decrease (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%) in the level of a biomarker of 8-hydroxydeoxyguanosine (8-OHdG), γ-H2AX, or DNA mutations (e.g., double strand breaks) in the subject.

[0112] Example 11 - Measuring Improvement of Nutritional Perception Disorders

[0113] The success of the compositions of the invention in improving dysregulated nutrient sensing in a subject (e.g., a human subject) can be measured, for example, by performing any of the following determinations: (i) an increase in the level of insulin sensitivity in a subject (e.g., an increase of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%); or (ii) a decrease in fasting glucose, fasting insulin, LDL, HbA1c, mTOR activity, or mitochondrial reactive oxygen species levels in a subject (e.g., a decrease of at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%).

[0114] The following content corresponds to the original claims in the parent application and is now incorporated herein as part of the specification:

[0115] 1. A composition comprising at least three substances selected from the group consisting of alpha-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger, Rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin, L-theanine, oxaloacetic acid, fumaric acid, succinic acid, hyaluronic acid, butyric acid, anthocyanins, piperamide, quercetin, curcuminoids, caffeine, trehalose, and spermidine.

[0116] 2. The composition of claim 1, wherein the composition comprises at least three substances selected from the group consisting of α-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger extract, rhodiola rosea, acetylglucosamine, vitamin C, glucosamine, fisetin, and L-theanine.

[0117] 3. The composition of claim 1, wherein the composition comprises alpha-ketoglutarate, microdose lithium, and fisetin.

[0118] 4. The composition of claim 3, wherein the composition comprises alpha-ketoglutarate, microdose lithium, fisetin, glycine and pterostilbene.

[0119] 5. The composition of claim 1, wherein the composition comprises alpha-ketoglutarate, microdose lithium, fisetin, glycine, pterostilbene, and glucosamine.

[0120] 6. The composition of claim 1, wherein the composition comprises α-ketoglutarate, fisetin, pterostilbene and glycine.

[0121] 7. The composition of claim 6, wherein the composition comprises α-ketoglutarate, fisetin, glycine, pterostilbene and glucosamine.

[0122] 8. The composition of claim 1, wherein the composition comprises alpha-ketoglutaric acid, malic acid, pterostilbene, microdose lithium, glycine, ginger extract, rhodiola rosea, hyaluronic acid, vitamin C, glucosamine, fisetin, and L-theanine.

[0123] 9. The composition of claim 8, wherein the composition comprises 1,100 mg calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin, and 150 mg L-theanine.

[0124] 10. The composition of claim 9, wherein the composition comprises 1,100 mg calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin, 150 mg L-theanine, xylitol, malic acid, flavoring agents, silicon dioxide, calcium silicate, and rebaudioside A.

[0125] 11. The composition of any one of items 1 to 10, wherein one or more of the substances in the composition are formulated for delayed release and / or extended release.

[0126] 12. The composition of claim 11, wherein the alpha-ketoglutarate, microdose lithium, fisetin, butyric acid, pterostilbene and glycine are formulated for delayed release and / or extended release.

[0127] 13. The composition of claim 11, wherein each substance is formulated for delayed release and / or extended release.

[0128] 14. A composition comprising (i) the composition of any one of items 1 to 13 and (ii) a pharmaceutically acceptable carrier.

[0129] 15. A method for improving at least one characteristic of aging in a subject, comprising administering to the subject the composition of any one of items 1 to 14, wherein the characteristic of aging is selected from genomic instability, telomere attrition, epigenetic changes, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

[0130] 16. The method of claim 15, wherein the method further comprises concurrently administering a nicotinamide-based compound to the subject.

[0131] 17. The method of claim 15 or 16, wherein the method improves genomic instability, telomere attrition, one or more epigenetic changes, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

[0132] 18. The method of any one of items 15 to 17, wherein the subject is a human.

[0133] 19. The method of any one of items 15 to 17, wherein the subject is a cat, a dog or a horse.

[0134] 20. An article comprising (i) the composition of any one of items 1 to 14 and (ii) a niacinamide-based compound in separate compartments.

[0135] 21. The article of claim 20, wherein the article comprises in separate compartments (i) a composition comprising alpha-ketoglutarate, malic acid, pterostilbene, microdose lithium, glycine, ginger extract, rhodiola rosea, hyaluronic acid, vitamin C, glucosamine, fisetin, and L-theanine, and (ii) nicotinamide mononucleotide.

[0136] 22. The article of claim 21, wherein the article comprises in separate compartments (i) a composition comprising 1,100 mg calcium alpha-ketoglutarate, 2,000 mg magnesium malate, 50 mg pterostilbene, 1 mg microdose lithium, 2,000 mg glycine, 80 mg ginger extract, 300 mg rhodiola rosea, 100 mg hyaluronic acid, 100 mg vitamin C, 1,000 mg glucosamine sulfate, 100 mg fisetin, and 150 mg L-theanine, and (ii) 125 mg nicotinamide mononucleotide.

[0137] References

[0138] 1.The Aging Process,Denham Harman.Proceedings of the National Academyof Sciences of the United States of America,1981.

[0139] 2.Mortality in random ized trials of antioxidant supplements forprimary and secondary prevention:systematic review and meta-analysis.JAMA,2007.

[0140] 3.Multivitamin-multimineral supplementation and mortality:a meta-analysis of randomized controlled trials.Am.J.Clin.Nutr.,2013.

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Claims

1. A composition comprising alpha-ketoglutarate, microdose lithium and fisetin.

2. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin, glycine and pterostilbene.

3. A composition comprising alpha-ketoglutarate, fisetin, pterostilbene and glycine.

4. A composition comprising alpha-ketoglutarate, fisetin, glycine, pterostilbene and glucosamine.

5. A composition comprising lithium, malic acid and alpha-ketoglutaric acid.

6. A composition comprising lithium, malic acid, alpha-ketoglutaric acid and vitamin C.

7. A composition comprising lithium, glycine, malic acid and alpha-ketoglutarate.

8. A composition comprising lithium, glycine, malic acid, alpha-ketoglutarate and vitamin C.

9. A composition comprising lithium, glycine, malic acid, fisetin and alpha-ketoglutarate.

10. A composition comprising lithium, glycine, malic acid, fisetin, alpha-ketoglutarate and vitamin C.

11. A composition comprising alpha-ketoglutarate, microdose lithium, and glucosamine.

12. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin and pterostilbene.

13. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin, and glucosamine.

14. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin, and glycine.

15. A composition comprising alpha-ketoglutarate, fisetin and spermidine.

16. A composition comprising alpha-ketoglutarate, fisetin, spermidine and pterostilbene.

17. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin and spermidine.

18. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin, and butyric acid.

19. A composition comprising fisetin, pterostilbene and glucosamine.

20. A composition comprising fisetin, glycine and pterostilbene.

21. A composition comprising fisetin, glucosamine and glycine.

22. A composition comprising pterostilbene, glycine and glucosamine.

23. A composition comprising fisetin, pterostilbene, glucosamine and glycine.

24. A composition comprising fisetin, pterostilbene, glucosamine, glycine and malic acid.

25. A composition comprising fisetin, pterostilbene and acetylglucosamine.

26. A composition comprising fisetin, pterostilbene and theanine.

27. A composition comprising alpha-ketoglutarate, fisetin and theanine.

28. A composition comprising alpha-ketoglutarate, microdose lithium, and theanine.

29. A composition comprising alpha-ketoglutarate, microdose lithium, fisetin, and theanine.

30. The composition of any one of claims 1 to 29, wherein each substance is formulated for delayed release and / or extended release.

31. A composition comprising (i) the composition of any one of claims 1 to 30 and (ii) a pharmaceutically acceptable carrier.

32. Use of the composition of any one of claims 1 to 31 in the preparation of a medicament for use in a method for ameliorating at least one characteristic of aging in a subject, the method comprising administering the composition to the subject, wherein the characteristic of aging is selected from genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

33. The use of claim 32, wherein the method ameliorates genomic instability, telomere attrition, one or more epigenetic alterations, loss of protein homeostasis, dysregulated nutrient sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.

34. The use of claim 32 or 33, wherein the subject is a human.

35. The use of claim 32 or 33, wherein the subject is a cat, dog or horse.