GLUTATHIONE COMPOSITIONS AND METHODS OF USE

PRIORITY

[0001 ] This application claims benefit of priority to U.S. Provisional Application No: 63/264,909 filed on December 03, 2021 , the contents of which are herein incorporated by reference in their entirety.

BACKGROUND

[0002] Many consequences of increased chronological age can be broadly attributed to an organism’s inability to maintain homeostasis at the cellular and organism level. Adaptive homeostasis is a phenomenon, shared across cells and tissues of both simple and complex organisms, that enables the expansion or contraction of the homeostatic range to modulate stress-protective systems in response to varying internal and external environments. As organisms age, the homeostasis can be disrupted due to an imbalance in the stress and stress protective mechanisms. This dysregulation of adaptive homeostasis in the final third of lifespan is a factor in the ageing process and stress associated disease, while successful maintenance or restoration of adaptive homeostasis can result in extended, healthy longevity.

SUMMARY

[0003] The present disclosure provides compositions and methods for use in treating a subject. In some embodiments the compositions can include glutathione, coenzyme Q10 (CoQ10), pyrroloquinoline quinone, iron binding glycoprotein or any combination thereof. In some embodiments, the concentration of glutathione can be about 10 mg/ml to 100 mg/ml. In some embodiments, the concentration of glutathione can be about 22 mg/ml. In some embodiments, the concentration of CoQ10 can be about 10 mg/ml to 100 mg/ml. In some embodiments, the concentration of CoQ10 can be about 22 mg/ml. In some embodiments, the concentration of pyrroloquinoline quinone can be about 1 mg to 10 mg. In some embodiments, the concentration of pyrroloquinoline quinone can be about 2 mg/ml. In some embodiments, the iron binding glycoprotein can be lactoferrin or transferrin. As a non-limiting example, the iron binding glycoprotein can be lactoferrin. In one embodiment, lactoferrin is a bovine lactoferrin. The concentration of bovine lactoferrin can be about 5 mg/ml to 50 mg/ml. In some embodiments, the concentration of bovine lactoferrin can be about 7 mg/ml.

[0004] Also provided herein are formulations comprising the compositions and at least one excipient. The excipient can be glycerin, oleic acid (from olive oil), natural clove oil, orange oil, cinnamon oil, vitamin E, milk and/or vanilla bean extract. In some embodiments, the formulation can be prepared as a liposome.

[0005] In some embodiments, the liposomes can include phosphatidylcholine. In some embodiments, the formulations can be present within a gel. The gel can include nut butter and/or an oil. The nut butter can be almond butter. In some embodiments, the oil can be medium chain triglyceride (MCT) oil and/or sunflower oil. As a non-limiting example, a gel can include almond butter, MCT oil and/or sunflower oil.

[0006] The present disclosure also provides methods of use. In some embodiments, the compositions can be used to treat Chronic Fatigue Syndrome (CFS). In some embodiments, the compositions can improve one or more symptoms associated with CFS. In some embodiments, the composition can be administered for about 50-150 days, for example, for about 30 days, 60 days, 90 days or more. Methods of treatment of diseases such as, but not limited to, cardiovascular diseases, diabetes, cancer, chronic kidney disease, chronic obstructive pulmonary disease (COPD), and/or neurological diseases are also provided herein.

[0007] The present disclosure also provides a method of increasing healthspan or lifespan in a subject by administering the compositions of the disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Various objectives, features, and advantages of the disclosed subject matter can be more fully appreciated with reference to the following detailed description of the disclosed subject matter when considered in connection with the following drawings, in which like reference numerals identify like elements.

[0009] Figure 1 shows adaptive responses in various age groups.

[0010] Figure 2 is an example of the formulation of the disclosure prepared as a liposome.

DETAILED DESCRIPTION

[0011 ] The details of one or more variations of the subject matter described herein are set forth in the accompanying drawings and the description below. Other features and advantages of the subject matter described herein will be apparent from the description and drawings, and from the claims. The disclosed subject matter is not, however, limited to any particular embodiment disclosed.

Overview

[0012] Reactive oxygen species (ROS) are produced by cells as normal cellular metabolic byproducts. Environmental toxins, age, stress, pharmaceutical agents, or pre-existing conditions can result in an increase in ROS. Excess reactive oxygen species (ROS) can lead to the destruction of biomolecules, and cellular organelles. Under excessive stress conditions, cells can produce numerous ROS, and the living organisms eventually evolve a series of response mechanisms to adapt to the ROS exposure as well as utilize it as the signaling molecules. ROS molecules can also trigger oxidative stress in a feedback mechanism involving many biological processes, such as apoptosis, necrosis and autophagy. Furthermore, ROS can play a role as the signaling molecules in the cell death pathway. Overwhelming production of ROS can destroy organelle’s structure and biomolecules, which also leads to inflammatory responses. Oxidative damage ensues when the generation of free radicals and reactive oxygen and nitrogen species exceeds the capacity of cellular defense mechanisms.

[0013] Organisms are equipped with mechanisms to combat ROS. As organisms age, there an imbalance between the systemic manifestation of ROS and the organism’s ability to readily detoxify ROS or to repair the resulting damage. This imbalance is referred to as oxidative stress. Mitochondria play a critical role in combatting ROS. However, mitochondrial biogenesis declines with age due to alterations in mitochondrial dynamics and inhibition of mitophagy, an autophagy process that removes dysfunctional mitochondria. Age-dependent abnormalities in mitochondrial quality control further weaken and impair mitochondrial function. In addition, with advanced chronological age, antioxidant concentrations in blood can decline leading to an increase in reactive oxygen species (ROS).

[0014] Excess reactive oxygen species (ROS) can lead to the destruction of biomolecules and cellular organelles which lead to chronic age-related conditions such as neurodegenerative diseases, cancer, diabetes, and kidney disease. In the case of neurodegenerative diseases, the nerve cells from brain and spinal cord can suffer from mitochondrial dysfunctions as well as exci to toxicity and finally apoptosis, which lead to either functional loss or sensory dysfunctions. Since the brain has a high metabolic rate and relatively reduced capacity for cellular regeneration, nerve cells in the brain area are particularly susceptible to damage by ROS. Long term, free radicals and oxidative stress can work together to accelerate aging and increase the risk of diseases e.g., fatigue, cancer, inflammatory joint disease, dementia, asthma and diabetes among others. There is a need in the art for compositions that can combat oxidative stress and prevent or reduce the incidence of diseases and conditions associated with increased oxidative stress.

[0015] The present disclosure provides compositions of glutathione (GSH), CoQ10, pyrroloquinoline quinone, and/or iron binding glycoproteins for restoring adaptive homeostasis by up-regulating aging related pathways and related gene expressions. In some embodiments, the compositions of the disclosure can reduce or prevent the decline of biological homeostasis and adaptability by upregulating detoxification pathways such as mitophagy, autophagy and ROS and down regulating mTOR, insulin and AMPK, while increasing the density and number of mitochondria and increasing the efficiency of their electron transport.

Glutathione

[0016] In some embodiments, the compositions of the disclosure can include glutathione In some embodiments, the glutathione is reduced glutathione (GSH). In the present disclosure the terms glutathione and reduced glutathione can be used interchangeably and are used to indicate the reduced form, GSH. Glutathione is a fundamental antioxidant that reduces ROS within the body, protecting cells and preventing tissue damage.

[0017] Glutathione (GSH) is an abundant antioxidant and a major detoxification agent in cells. In some embodiments, GSH can be synthesized by the sequential addition of the precursor amino acids, cysteine (Cys), glutamic acid (Glu) and glycine (Gly) through two- enzyme reactions catalyzed by glutamate cysteine ligase and glutathione synthetase. GSH levels are regulated in response to redox change. It plays a role in cell growth, proliferation, apoptosis and signaling, particularly redox signaling. GSH acts as an antioxidant in two ways, it directly reacts non-enzymatically with free radicals such as superoxide radical (O2"), NO, or hydroxyl ion (OH") and it indirectly functions as a reducing agent by donating an electron to H2O2 for its reduction to water and O2 in the presence of glutathione peroxidase (GPx) enzyme. Due to its potential cytoprotective functions there has been great interest in increasing GSH via analogs and precursors. Accordingly, GSH can play a role in protecting neuronal tissue, pulmonary cells and cardiovascular structures by enhancing GSH-related detoxification capacity.

[0018] Low GSH levels can impair immune function. GSH is also involved in the maintenance of optimal cytokine levels. GSH depletion can lead to an increase in pro- inflammatory cytokines i.e., tumor necrosis factor-a (TNF-a), lnterleukin-1 p (IL-1 p) and IL-6 gene expression but may not affect levels of anti-inflammatory cytokines such as IL-10. Low levels of GSH can be associated with exposure to oxidants/drugs/toxins, poor nutrition and other factors. GSH levels are also dependent on the availability of its precursor amino acids, Cys, Glu and Gly, with Cys most often considered as rate limiting. Mouse models have suggested that intracellular GSH levels can be depleted in certain tissues, including the liver, by short periods of fasting such as that which occurs overnight. Additionally, GSH is the most abundant brain antioxidant and altered GSH can be a key aspect of the pathogenesis of mild cognitive impairment, many neurological disorders and Alzheimer’s disease. Furthermore, higher GSH levels have been identified in the brains of older adults compared to their younger counterparts, suggesting that the higher GSH levels are present to help offset the age-related increases in oxidative stress and therefore maintain redox homeostasis.

CoQ10

[0019] In some embodiments, compositions of the disclosure can include CoQ10. CoQ10 can improve the speed and efficiency of mitochondria facilitating more energy transfer and energy availability at a cellular level.

[0020] Coenzyme Q10 (CoQ10), also known as ubiquinone, is a lipid-soluble vitamin-like compound. CoQ10 is present in the inner membrane of the mitochondria of the cell. High levels of CoQ10 is found in high energy cells such as heart, brain, liver, and kidney cells. In the aging process, cellular production of CoQ10 decreases. [0021 ] There are at least two functions of CoQ10. First, CoQ10 acts as a regenerating antioxidant, helping to quench free radicals and prevent lipid peroxidation. Second, CoQ10 is a mitochondrial energy coupler, which is part of the cellular process for synthesizing adenosine triphosphate (ATP), the body’s main source of energy.

[0022] By facilitating ATP synthesis in the mitochondria and upregulating mitochondrial function, CoQ10 can also support cognitive function. CoQIO’s antioxidant activity also has neuroprotective effects by protecting against oxidative stress in the mitochondria of brain cells. CoQ10 can also attenuate oxidative damage in the mitochondria to reduce cognitive effects associated with normal brain aging, and to supplement spatial learning.

[0023] CoQ10 can decrease hepatic mRNA expression of IL-6 and TNF-a, to attenuate the level of C-reactive protein (CRP) in rodent models. CoQ10 supplementation with doses ranging from about 100-500 mg/day can increase the serum level of CoQ10, and decrease TNF-a, but no substantial difference is observed in CRP.

[0024] CoQ10 oral administration can provide beneficial effects on a range of different disorders that have been associated with low CoQ10 levels and high oxidative stress, such as, but not limited to, mitochondrial, cardiovascular, and neurodegenerative diseases.

Pyrroloquinoline quinone

[0025] In some embodiments, compositions of the disclosure can include pyrroloquinoline quinone (PQQ), an aromatic tricyclic o-quinone that serves as a redox cofactor of a number of prokaryotic dehydrogenases, such as alcohol and sugar dehydrogenases. PQQ can exert pro-oxidant actions by the formation of reactive oxygen species (ROS), such as O2- and H2O2, via its redox cycling under certain conditions. PQQ can influence energy-related metabolism and neurologic functions in animals. The mechanism of action involves interactions with cell signaling pathways and mitochondrial function. PQQ has anti-oxidative and mitochondrial biogenesis capacities. Reduced PQQ can provide strong antioxidant capacity. In some embodiments, the antioxidant capacity can be greater than ascorbic acid, by about 2 times, 3 times, 4 times, 5 times, 6 times, 7, times, 8 times, 9 times or 10 times. As a non-limiting example, the antioxidant capacity of the compositions of the disclosure can be about 7.4 times greater than ascorbic acid. Increasing PQQ levels can trigger the growth of new mitochondria and nerve cells with strong neuroprotective signals.

Iron Binding Glycoprotein [0026] In some embodiments, the compositions of the disclosure can include iron binding glycoproteins. In some embodiments, the iron binding glycoprotein is used for absorption via the jejunum (the second part of the small intestine).

[0027] The iron binding glycoprotein can be a synthetic iron binding glycoprotein, a recombinant iron binding glycoprotein or a natural iron binding glycoprotein. The iron binding glycoprotein can be derived from any mammal. In some embodiments, the iron binding glycoprotein can be derived from a cow, human, monkey, dog, cat, mouse, rat, guinea pig, and/or hamster.

[0028] In some embodiments, the iron binding glycoprotein can be lactoferrin or transferrin.

[0029] In some embodiments, the iron binding glycoprotein can be lactoferrin. Lactoferrin (LF) is a cationic iron-binding glycoprotein with molecular weight of about 77-80 kDa. Lactoferrin is folded into two globular lobes connected by an a-helical bridge and stabilized by a number of disulfide bonds. The main function of this protein is to control and regulate the concentration of free iron in biological fluids via its ability to either solubilize or sequester ferric ions (Fe ³⁺ ). LF can be either extracted from milk or produced by recombinant techniques in microorganisms for use in the compositions of the disclosure. Generally, one LF molecule can typically binds two Fe ³⁺ ions, which greatly depends on the synergistic binding of two HCOs " or CO3 ² ". In vitro iron release can be enhanced from LF molecule by lowering the pH or using chelating agents. Moreover, structure analysis of LF revealed that it is composed of 2 lobes. Each lobe includes 2 domains separated by a cleft that can cobind Fe ³⁺ and CO3 ² ". The binding and the release of substrates is usually associated with conformational changes including opening and closing of the interdomain cleft. Two carbohydrate attachment sites are present on the LF molecule, one in each lobe. The first [3- a-p unit (residues 1 to 40) includes about nine basic sidechains protruding from the surface (most near the C terminus of helix A) which seems to be a receptor binding moiety, responsible for receptor recognition of LF.

[0030] In some embodiments, lactoferrin can be bovine lactoferrin (UniProt ID: Q6LBN7). [0031 ] In some embodiments, the lactoferrin can be ENSEMBL translation ID ENSP00000231751.4, ENSP00000395234.1 , ENSP00000397427.1 , ENSP00000405546.1 , ENSP00000405719.2, ENSP00000508000.1 , a portion, or a variant thereof. [0032] As a non-limiting example, the iron binding glycoprotein can be transferrin. Presence of transferrin in the compositions can allow the safe transfer of the compositions through the gastric environment. Human transferrin is a single-chain glycoprotein. It has two potential N-linked glycosylation sites and one potential O-linked glycosylation site. Transferrin binds two Fe ³⁺ ions, with high affinity (Kd ~ 10-20 M), together with two synergistically bound COs ^2- ions. The protein is mainly synthesized in the liver and secreted into the plasma. Lower amounts are also produced in other organs, such as the testis and brain. Transferrin can also act as a cytokine. For example, transferrin has been identified as one of the factors that promote the clonal growth of the precursors of murine granulocytes and macrophages cultured in vitro under serum-free conditions. In some embodiments, the compositions of the disclosure can include the entire transferrin protein or a fragment thereof. In some embodiments, the transferrin can be ENSEMBL Translation ID ENSP00000385834.3, ENSP00000419338.1 , ENSP00000417468.1 , a portion thereof or a variant thereof. Physiologically, transferrin is responsible for the transport of iron and controls the level of iron in biological fluids. Transferrin-receptors (TfR) on cell surfaces, can bind Tf with high affinity and internalize into the cell. Transferrin receptor (TfR) can also transport transferrin across the blood-brain barrier (BBB), which can be exploited to deliver the compositions of the disclosure from the bloodstream into the brain.

[0033] In some embodiments, transferrin can be bovine transferrin or human transferrin. [0034] In some aspects an iron binding protein can be covalently or non-covalently linked to glutathione, coenzyme Q10 (CoQ10), pyrroloquinoline quinone, or combinations thereof. Compositions

[0035] Compositions of the disclosure can include a combination comprising one or more of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein. Compositions can be prepared in an oxygen deprived environment to prevent oxidation of the composition.

[0036] In some embodiments, the compositions can include glutathione in the amount of from about 0.1 mg to 1 mg, about 0.5 mg to 5 mg, 1 mg to 10 mg, 5 mg to 50 mg, 10 mg to 100 mg, 50 mg to 150 mg, 100 mg to 200 mg, 150 mg to 250 mg, 200 mg to 300 mg, 250 mg to 350 mg, 300 mg to 400 mg, 350 mg to 450 mg, 400 mg to 500 mg, 450 mg to 550 mg, 500 mg to 600 mg, 550 mg to 650 mg, 600 mg to 700 mg, 650 mg to 750 mg, 700 mg to 800 mg, 850 mg to 950 mg, 900 mg to 1000 mg or more. [0037] In some embodiments, the compositions can include glutathione in the amount of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, or more. In some embodiments, the compositions can include glutathione in the amount of 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, or 300 mg.

[0038] In some embodiments, the compositions can include glutathione in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. [0039] In some embodiments, the compositions can include glutathione in the amount of 10 mg/ml, 20 mg/ml, 30 mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, 90 mg/ml, 100 mg/ml, 200 mg/ml, 300 mg/ml, 400 mg/ml, 500 mg/ml, 600 mg/ml, 700 mg/ml, 800 mg/ml, or more. In some embodiments, the compositions can include glutathione in the amount of 15 mg/ml, 16 mg/ml, 17 mg/ml, 18 mg/ml, 19 mg/ml, 20 mg/ml, 21 mg/ml, 22 mg/ml, 23 mg/ml, 24 mg/ml, 25 mg/ml, 26 mg/ml, 27 mg/ml, 28 mg/ml, 29 mg/ml, 30 mg/ml, 31 mg/ml, 32 mg/ml, 33 mg/ml, 34 mg/ml, 35 mg/ml, 36 mg/ml, 37 mg/ml, 38 mg/ml, 39 mg/ml, or 40 mg/ml.

[0040] In some embodiments, the compositions can include CoQ10 in the amount of from about 0.1 mg to 1 mg, about 0.5 mg to 5 mg, 1 mg to 10 mg, 5 mg to 50 mg, 10 mg to 100 mg, 50 mg to 150 mg, 100 mg to 200 mg, 150 mg to 250 mg, 200 mg to 300 mg, 250 mg to 350 mg, 300 mg to 400 mg, 350 mg to 450 mg, 400 mg to 500 mg, 450 mg to 550 mg, 500 mg to 600 mg, 550 mg to 650 mg, 600 mg to 700 mg, 650 mg to 750 mg, 700 mg to 800 mg, 850 mg to 950 mg, 900 mg to 1000 mg or more.

[0041 ] In some embodiments, the compositions can include CoQ10 in the amount of 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, or more. In some embodiments, the compositions can include CoQ10 in the amount of 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, or 300 mg. [0042] In some embodiments, the compositions can include CoQ10 in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. [0043] In some embodiments, the compositions can include CoQ10 in the amount of 10 mg/ml, 20 mg/ml, 30 mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, 90 mg/ml, 100 mg/ml, 200 mg/ml, 300 mg/ml, 400 mg/ml, 500 mg/ml, 600 mg/ml, 700 mg/ml, 800 mg/ml, or more. In some embodiments, the compositions can include CoQ10 in the amount of 15 mg/ml, 16 mg/ml, 17 mg/ml, 18 mg/ml, 19 mg/ml, 20 mg/ml, 21 mg/ml, 22 mg/ml, 23 mg/ml, 24 mg/ml, 25 mg/ml, 26 mg/ml, 27 mg/ml, 28 mg/ml, 29 mg/ml, 30 mg/ml, 31 mg/ml, 32 mg/ml, 33 mg/ml, 34 mg/ml, 35 mg/ml, 36 mg/ml, 37 mg/ml, 38 mg/ml, 39 mg/ml, or 40 mg/ml. [0044] In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of from about 0.1 mg to 1 mg, about 0.5 mg to 5 mg, 1 mg to 10 mg, 5 mg to 50 mg, 10 mg to 100 mg, 50 mg to 150 mg, 100 mg to 200 mg, 150 mg to 250 mg, 200 mg to 300 mg, 250 mg to 350 mg, 300 mg to 400 mg, 350 mg to 450 mg, 400 mg to 500 mg, 450 mg to 550 mg, 500 mg to 600 mg, 550 mg to 650 mg, 600 mg to 700 mg, 650 mg to 750 mg, 700 mg to 800 mg, 850 mg to 950 mg, 900 mg to 1000 mg or more.

[0045] In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, or more. In some embodiments, the compositions can include CoQ10 in the amount of 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, or 30 mg.

[0046] In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. [0047] In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of 1 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 20 mg/ml, 30 mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, or more. In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of 1.5 mg/ml, 1.6 mg/ml, 1.7 mg/ml, 1.8 mg/ml, 1.9 mg/ml, 2.0 mg/ml, 2.1 mg/ml, 2.2 mg/ml, 2.3 mg/ml, 2.4 mg/ml, 2.5 mg/ml, 2.6 mg/ml, 2.7 mg/ml, 2.8 mg/ml, 2.9 mg/ml, 3.0 mg/ml, 3.1 mg/ml, 3.2 mg/ml, 3.3 mg/ml, 3.4 mg/ml, 3.5 mg/ml, 3.6 mg/ml, 3.7 mg/ml, 3.8 mg/ml, 3.9 mg/ml, or 4.0 mg/ml.

[0048] In some embodiments, the compositions can include iron binding glycoprotein in the amount of from about 0.1 mg to 1 mg, about 0.5 mg to 5 mg, 1 mg to 10 mg, 5 mg to 50 mg, 10 mg to 100 mg, 50 mg to 150 mg, 100 mg to 200 mg, 150 mg to 250 mg, 200 mg to 300 mg, 250 mg to 350 mg, 300 mg to 400 mg, 350 mg to 450 mg, 400 mg to 500 mg, 450 mg to 550 mg, 500 mg to 600 mg, 550 mg to 650 mg, 600 mg to 700 mg, 650 mg to 750 mg, 700 mg to 800 mg, 850 mg to 950 mg, 900 mg to 1000 mg or more.

[0049] In some embodiments, the compositions can include iron binding glycoprotein in the amount of 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, or more. In some embodiments, the compositions can include iron binding glycoprotein in the amount of 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, or 80 mg.

[0050] In some embodiments, the compositions can include iron binding glycoprotein in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more.

[0051 ] In some embodiments, the compositions can include iron binding glycoprotein in the amount of 1 mg/ml, 2 mg/ml, 3 mg/ml, 4 mg/ml, 5 mg/ml, 6 mg/ml, 7 mg/ml, 8 mg/ml, 9 mg/ml, 10 mg/ml, 20 mg/ml, 30 mg/ml, 40 mg/ml, 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, or more. In some embodiments, the compositions can include pyrroloquinoline quinone in the amount of 5 mg/ml, 5.5 mg/ml, 6 mg/ml, 6.5 mg/ml, 7 mg/ml, 7.5 mg/ml, 8 mg/ml, 8.5 mg/ml, 9 mg/ml, 9.5 mg/ml, 10 mg/ml, 10.5 mg/ml, 11 mg/ml, 11.5 mg/ml, 12 mg/ml, 12.5 mg/ml, 13 mg/ml, 13.5 mg/ml, 14 mg/ml, 14.5 mg/ml, 15 mg/ml, 15.5 mg/ml, 16 mg/ml, 16.5 mg/ml, 17 mg/ml, 17.5 mg/ml, 18 mg/ml, 18.5 mg/ml, 19 mg/ml, 19.5 mg/ml or more.

[0052] As a non-limiting example, the compositions of the disclosure can include the following components: glutathione (about 100, 200, 220, 300, or 400 mg); CoQ10 (about 100, 200, 220, 300, or400 mg), pyrroloquinoline quinone (about 5, 10, 15, 20, 30, or40 mg) and/or iron binding glycoprotein e.g., lactoferrin (about 40, 50, 60, 70, 80, 90, or 100 mg). In some embodiments, the compositions can include glutathione (about 15, 20, 22, 25, 30, or 40 mg/ml); CoQ10 (about 15, 20, 22, 25, 30 or 40 mg/ml), pyrroloquinoline quinone (about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, or 3.5 mg/ml) and/or iron binding glycoprotein e.g., lactoferrin (about 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/ml).

[0053] As a non-limiting example, the compositions of the disclosure can include the following components: glutathione (about 220 mg); CoQ10 (about 220 mg), pyrroloquinoline quinone (about 20 mg) and/or iron binding glycoprotein e.g., lactoferrin (about 70 mg). In some embodiments, the compositions can include glutathione (about 22 mg/ml); CoQ10 (about 22 mg/ml), pyrroloquinoline quinone (about 2 mg/ml) and/or iron binding glycoprotein e.g., lactoferrin (about 7 mg/ml).

Formulations

[0054] In some embodiments, the compositions can be administered to humans, human patients or subjects. Although the descriptions of formulations provided herein are principally directed to formulations which are suitable for administration to humans, it will be understood by the skilled artisan that such therapeutic agents are generally suitable for administration to any other animal, e.g., to non-human animals, e.g., non-human mammals. Modifications of formulations can be used such that they are suitable for administration to humans.

[0055] Formulations of compositions of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein described herein can be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein into association with an excipient and/or one or more other accessory ingredients, and then, if necessary and/or desirable, dividing, shaping and/or packaging the product into a desired single- or multi-dose unit. As a non-limiting example, the single-dose unit can be 10 ml. [0056] A formulation in accordance with the disclosure can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the formulation comprising a predetermined amount of the compositions of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein. The amount of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein is generally equal to the dosage which would be administered to a subject and/or a convenient fraction of such a dosage such as, for example, one-half or one- third of such a dosage.

[0057] Relative amounts of one or more of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein, the excipients, and/or any additional ingredients in a formulation in accordance with the disclosure can vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the formulation is to be administered. By way of example, the formulation can include between 0.1 % and 100%, e.g., between .5 and 50%, between 1-30%, between 5-80%, or, in some embodiments, at least 20%, at least 40%, at least 60%, or at least 80% (w/w) of one or more of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein.

[0058] The compositions of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein of the present disclosure can be formulated using one or more excipients to: (1) increase stability; (2) permit the sustained or delayed release; (3) alter the biodistribution; (4) alter the release profile of glutathione, CoQ10, pyrroloquinoline quinone and/or iron binding glycoprotein in vivo.

[0059] In one embodiment, the compositions are formulated to safely pass through the gastric environment.

Excipients

[0060] Formulations can additionally comprise an excipient, which, as used herein, includes any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, and the like, as suited to the particular dosage form desired. Remington’s The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference in its entirety) discloses various excipients used in preparing formulations and known techniques for the preparation thereof. Except insofar as any conventional excipient medium is incompatible with a substance or its derivatives, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the formulation, its use is contemplated to be within the scope of this disclosure.

[0061 ] In some embodiments, an excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, an excipient is approved for use in humans and for veterinary use. In some embodiments, an excipient is approved by United States Food and Drug Administration. In some embodiments, an excipient is pharmaceutical grade. In some embodiments, an excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the International Pharmacopoeia.

[0062] Formulations can include excipients, such as without limitation, binders, coating, disintegrants, fillers, diluents, flavors, colors, lubricants, glidants, preservatives, sorbents, sweeteners, conjugated linoleic acid (CLA), gelatin, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, oils, beeswax, purified water, glycerol, any type of oil, including, without limitation, fish oil or soybean oil, or the like.

[0063] Formulations can comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as, e.g., polyethylene glycols.

[0064] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, etc., and/or combinations thereof.

[0065] Exemplary granulating and/or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponge, cationexchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked polyvinylpyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (VEEGUM®), sodium lauryl sulfate, quaternary ammonium compounds, etc., and/or combinations thereof.

[0066] Exemplary surface active agents and/or emulsifiers include, but are not limited to, natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and VEEGUM® [magnesium aluminum silicate]), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g., carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [TWEEN®20], polyoxyethylene sorbitan [TWEENn®60], polyoxyethylene sorbitan monooleate [TWEEN®80], sorbitan monopalmitate [SPAN®40], sorbitan monostearate [SPAN®60], sorbitan tristearate [SPAN®65], glyceryl monooleate, sorbitan monooleate [SPAN®80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [ YRJ®45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and SOLUTOL®), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., CREMOPHOR®), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether [BRIJ®30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, PLUORINC®F 68, POLOXAMER®188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, etc. and/or combinations thereof.

[0067] Exemplary binding agents include, but are not limited to, starch (e.g., cornstarch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol,); natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, polyvinylpyrrolidone), magnesium aluminum silicate (Veegum®), and larch arabogalactan); alginates; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethacrylates; waxes; water; alcohol; etc.; and combinations thereof.

[0068] Exemplary preservatives can include, but are not limited to, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and/or other preservatives. Preservatives can be used to prevent spoilage from bacteria, molds, fungi, or yeast (antimicrobials); slow or prevent changes in color, flavor, or texture and delay rancidity (antioxidants); and/or maintain freshness. Exemplary antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and/or thimerosal. Exemplary antifungal preservatives include, but are not limited to, butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and/or sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and/or phenylethyl alcohol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and/or phytic acid. Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, GLYDANT PLUS®, PHENONIP®, methylparaben, GERMALL®115, GERMABEN®II, NEOLONE™ , KATHON™ , and/or EUXYL®.

[0069] Exemplary buffering agents include, but are not limited to, citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer’s solution, ethyl alcohol, etc., and/or combinations thereof.

[0070] Exemplary oils include, but are not limited to, almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oils. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and/or combinations thereof.

[0071 ] In some embodiments, the formulations can include almond butter, acorn butter, cashew butter, hazelnut butter, macadamia nut butter, peanut butter, pecan butter, pistachio butter, walnut butter, pumpkin seed butter, sesame seed butter (tahini), soybean butter, and/or sunflower seed butter. As a non-limiting example, the formulation can include about 5- 25% almond butter.

[0072] In some embodiments, the formulations can include sweeteners. These can be used to add sweetness with or without the extra calories. Nonlimiting examples of sweeteners include sucrose (sugar), glucose, fructose, sorbitol, mannitol, corn syrup, high fructose com syrup, saccharin, aspartame, sucralose, acesulfame potassium (acesulfame-K), and/or neotame.

[0073] In some embodiments, the formulations can include color additives. These can be used to offset color loss due to exposure to light, air, temperature extremes, moisture and storage conditions; correct natural variations in color; enhance colors that occur naturally; and/or to provide color to colorless foods. Nonlimiting examples of color additives include FD&C Blue Nos. 1 and 2, FD&C Green No. 3, FD&C Red Nos. 3 and 40, FD&C Yellow Nos. 5 and 6, Orange B, Citrus Red No. 2, annatto extract, beta-carotene, grape skin extract, cochineal extract or carmine, paprika oleoresin, caramel color, fruit and vegetable juices, and/or saffron. In some embodiments, the formulations can be dark orange to brown in color. [0074] In some embodiments, the formulations can include flavors and spices. These can be used to add specific flavors (natural and synthetic). Nonlimiting examples include natural flavoring, artificial flavor, and spices e.g., orange, cinnamon, vanillin, vanilla extract, mango extract, cinnamon, citrus, coconut, ginger, viridiflorol, almond, menthol (including menthol without mint), grape skin extract, and/or grape seed extract.

[0075] In some embodiments, the formulations can include flavor enhancers. These can be used to enhance flavors already present in foods (without providing their own separate flavor). Nonlimiting examples of flavor enhancers include monosodium glutamate (MSG), hydrolyzed soy protein, autolyzed yeast extract, disodium guanylate or inosinate.

[0076] In some embodiments, the formulations can include nutrients. These can be used to replace vitamins and minerals lost in processing (enrichment), add nutrients that can be lacking in the diet (fortification). Nonlimiting examples of nutrients include thiamine hydrochloride, riboflavin (Vitamin B2), niacin, niacinamide, folate or folic acid, beta carotene, potassium iodide, iron or ferrous sulfate, alpha tocopherols, ascorbic acid, Vitamin D, amino acids (L-tryptophan, L-lysine, L-leucine, L-methionine).

[0077] In some embodiments, the formulations can include stabilizers and thickeners, binders, and/or texturizers. These can be used to produce uniform texture, improve "mouthfeel." Non-limiting examples include gelatin, pectin, guar gum, carrageenan, xanthan gum, whey.

[0078] In some embodiments, the formulations can include pH control agents and acidulants. These can be used to control acidity and alkalinity, prevent spoilage. Nonlimiting examples include lactic acid, citric acid, ammonium hydroxide, or sodium carbonate.

[0079] In some embodiments, the formulations can include humectants. These can be used to retain moisture. Nonlimiting examples of humectants include glycerin, or sorbitol.

[0080] In some embodiments, the formulations of the disclosure can include gelling ingredients such as, but not limited to gelatin, alginate, carageenan, gum, pectin, konjac, agar, food acid, rennet, starch, starch derivatives, and/or combinations thereof.

[0081 ] In some embodiments, the formulations described herein can include one or more of natural almond butter, glycerin, medium chain triglycerides (MCT oil), oleic acid (from olive oil), phosphatidylcholine, natural clove oil, orange oil, cinnamon oil, vitamin E (as D alpha tocopherol), milk and/or vanilla bean extract.

[0082] Depending on the intended mode of administration, the formulation used can be in the form of solid, semi-solid or liquid dosage forms, such as, for example, tablets, suppositories, pills, capsules, powders, liquids, suspensions or the like, preferably in forms of unit dosage suitable for individual administration of precise dosages.

[0083] The formulation can be in the form of a gel, a tablet, a capsule, a liquid, an aerosol, a powder, an effervescent tablet or powder, a syrup, an emulsion, a suspension, a solution, or any other form for providing the formulation to a subject. In particular embodiments, the formulation can be in a form for oral administration, buccal administration, sublingual administration, or any other route of administration as known in the art.

[0084] In some embodiments, the nitrogen gas is utilized to remove oxygen from the formulation prior to packaging.

Liposomes

[0085] In some embodiments, the formulations of the disclosure are prepared as liposomes. A liposome is a microscopic structure made of one or more layers of phospholipid materials resembling the phospholipid that make up cell membranes. The outer portion of the liposome can be fat soluble, while the inner portion of the liposome can be water-soluble. [0086] Liposomes can include lipids having two hydrocarbon chains, typically acyl chains, and a head group, either polar or nonpolar. The hydrocarbon chains can be saturated or have varying degrees of unsaturation. The two hydrocarbon chains of the lipid can be symmetrical or asymmetrical. The acyl chains of the lipids can be saturated or unsaturated. Non limiting examples of saturated acyl groups include, trianoic, tetranoic, pentanoic, hexanoic, heptanoic, octanoic, nonanoic, decanoic, undecanoic, dodecanoic, tridecanoic, tetradecanoic, pentadecanoic, hexadecanoic, 3,7,11 ,15-tetramethylhexadecanoic, heptadecanoic, octadecanoic, nonadecanoic, eicosanoic, heneicosanoic, docosanoic, trocosanoic and tetracosanoic. Non-limiting examples of unsaturated acyl groups include 9- cis-tetradecanoic, 9-cis-hexadecanoic, 9-cis-octadecanoic, 9-trans-octadecanoic, 9-cis-12- cis-octadecadienoic, 9-cis-12-cis-15-cis-octadecatrienoic, 11-cis-eicosenoic and 5-cis-8-cis- 11 -cis-14-cis-eicosatetraenoic. [0087] Liposomes can include lipids, such as, but not limited to, sphingolipids, ether lipids, sterols, phospholipids, particularly the phosphoglycerides, and the glycolipids, such as the cerebrosides and/or gangliosides.

[0088] In some embodiments, the liposomes can contain phosphoglycerides. Phosphoglycerides include phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine phosphatidylglycerol and diphosphatidylglycerol (cardiolipin), where the two hydrocarbon chains are typically between about 14-22 carbon atoms in length and have varying degrees of unsaturation.

[0089] In some embodiments, the liposomes can contain phosphatidylcholines. Exemplary phosphatidylcholines include dilauroyl phophatidylcholine, dimyristoylphophatidylcholine, dipalmitoylphophatidylcholine, distearoylphophatidyl-choline, diarachidoylphophatidylcholine, dioleoylphophatidylcholine, dilinoleoyl-phophatidylcholine, dierucoylphophatidylcholine, palmitoyl-oleoyl-phophatidylcholine, egg phosphatidylcholine, myristoyl-palmitoylphosphatidylcholine, palmitoyl-myristoyl-phosphatidylcholine, myristoylstearoylphosphatidylcholine, palmitoyl-stearoyl-phosphatidylcholine, stearoylpalmitoylphosphatidylcholine, stearoyl-oleoyl-phosphatidylcholine, stearoyl- linoleoylphosphatidylcholine and palmitoyl-linoleoyl-phosphatidylcholine. Asymmetric phosphatidylcholines are referred to as 1-acyl, 2-acyl-sn-glycero-3-phosphocholines, wherein the acyl groups are different from each other. Symmetric phosphatidylcholines are referred to as 1,2-diacyl-sn-glycero-3-phosphocholines. As used herein, the abbreviation “PC” refers to phosphatidylcholine. The phosphatidylcholine 1 ,2-dimyristoyl-sn-glycero-3- phosphocholine is abbreviated herein as “DMPC.” The phosphatidylcholine 1 ,2-dioleoyl-sn- glycero-3-phosphocholine is abbreviated herein as “DOPC.” The phosphatidylcholine 1 ,2- dipalmitoyl-sn-glycero-3-phosphocholine is abbreviated herein as “DPPC.” Liposomes can include “caged” phospholipids. Caged phospholipids are amino phospholipids that when present in a liposome, render the liposome pH-sensitive so that once endocytosed into target cells the caging groups are cleaved. This cleavage results in destabilization of the liposome, causing the uncaged lipids of the liposome to become fusogenic and to thereby release of the active agent (e.g., compositions described herein) carried by the liposome into the cell. [0090] In some embodiments, the liposomes can contain phosphatidylethanolamines. Exemplary phosphatidylethanolamines include dimyristoyl-phosphatidylethanolamine, dipalmitoyl-phosphatidylethanolamine, distearoyl-phosphatidylethanolamine, dioleoylphosphatidylethanolamine and egg phosphatidylethanolamine.

[0091 ] In some embodiments, the liposomes can contain phosphatidic acids. Exemplary phosphatidic acids include dimyristoyl phosphatidic acid, dipalmitoyl phosphatidic acid and dioleoyl phosphatidic acid.

[0092] In some embodiments, the liposomes can contain phosphatidylserines. Exemplary phosphatidylserines include dimyristoyl phosphatidylserine, dipalmitoyl phosphatidylserine, dioleoylphosphatidylserine, distearoyl phosphatidylserine, palmitoyl-oleylphosphatidylserine and brain phosphatidylserine.

[0093] In some embodiments, the liposomes can contain phosphatidylglycerols. Exemplary phosphatidylglycerols include dilauryloylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, dioleoyl- phosphatidylglycerol, dimyristoylphosphatidylglycerol, palmitoyl-oleoyl-phosphatidylglycerol and egg phosphatidylglycerol. The phosphatidylglycerol 1 ,2-dimyristoyl-sn-glycero-3- [phospho-rac-(1 -glycerol)] is abbreviated herein as “DMPG”. The phosphatidylglycerol 1 ,2- dipalmitoyl-sn-glycero-3-(phospho-rac-1 -glycerol) (sodium salt) is abbreviated herein as “DPPG”.

[0094] In some embodiments, the liposomes can contain sphingomyelins. Suitable sphingomyelins can include brain sphingomyelin, egg sphingomyelin, dipalmitoyl sphingomyelin, and distearoyl sphingomyelin.

[0095] Other suitable lipids for use in liposomes include glycolipids, sphingolipids, ether lipids, glycolipids such as the cerebrosides and gangliosides, and sterols, such as cholesterol or ergosterol.

[0096] The overall surface charge of the liposome can affect the tissue uptake of a liposome. In certain embodiments, anionic phospholipids such as phosphatidylserine, phosphatidylinositol, phosphatidic acid, and cardiolipin are used In some embodiments, liposome can include neutral lipids such as dioleoylphosphatidyl ethanolamine (DOPE). In some embodiments, cationic lipids can contain a lipophilic moiety, such as a sterol, an acyl or diacyl chain, and where the lipid has an overall net positive charge.

[0097] Liposomes containing formulations of the disclosure having a more rigid lipid bilayer, or a liquid crystalline bilayer, are achieved by incorporation of a relatively rigid lipid. The rigidity of the lipid bilayer correlates with the phase transition temperature of the lipids present in the bilayer. Phase transition temperature is the temperature at which the lipid changes physical state and shifts from an ordered gel phase to a disordered liquid crystalline phase. Several factors affect the phase transition temperature of a lipid including hydrocarbon chain length and degree of unsaturation, charge and headgroup species of the lipid. Lipid having a relatively high phase transition temperature can be used to produce a more rigid bilayer. Other lipid components, such as cholesterol, are also known to contribute to membrane rigidity in lipid bilayer structures.

[0098] In some embodiments, the liposomes can be hollow microspheres of different sizes, ranging from 50nm to 1000nm, made up of one or more lipid bilayers that enclose a hydrophilic core. In an aqueous medium, hydrophobic tails attract each other, while hydrophilic heads tend to face water. The result is lipid bilayers that close to form small vesicles within which there is a varied hydrophilic environment. In some embodiments, the hydrophilic core can include glutathione (GSH), pyrroloquinoline quinone, and/or iron binding glycoprotein. In some embodiments, the CoQ10 of the composition can be associated with a hydrophobic portion of the liposomes, e.g., the hydrophobic tails of lipids making up the liposomes, a lipid bilayer of the liposomes, or a lipid membrane of the liposomes.

[0099] Liposomes can be classified according to their size and number of lipid bilayers. Liposomes can be multilamellar liposome vesicles: they have an onion-like structure in which various lipid bilayers are intercalated with hydrophilic layers; unilamellar liposomes vesicles formed by a single lipid bilayer and enclose a strongly hydrophilic nucleus or oligolamellar vesicles, made up of several lipid bilayers that enclose a markedly hydrophobic environment. [0100] In one embodiment, the liposomes, can be, multilamellar vesicles (MLV), microemulsified liposomes (MEL), or large unilamellar vesicles (LUVET).

[0101 ] In some embodiments, the liposomes can have a length or diameter of about 1 - 10 nm, 5 - 15 nm, 10 - 20 nm, 15 - 25 nm, 20 - 30 nm, 25 - 35 nm, 30 - 40 nm, 35 - 45 nm, 40 - 50 nm, 45 - 55 nm, 50 - 60 nm, 55 - 65 nm, 60 - 70 nm, 65 - 75 nm, 70 - 80 nm, 75 - 85 nm, 80 - 90 nm, 85 - 95 nm, 90 - 100 nm, 100 - 200 nm, 150 - 250 nm, 200 - 300 nm, 250 - 350 nm, 300 - 400 nm, 350 - 450 nm, 400 - 500 nm, 450 - 550 nm, 500 - 600 nm, 550 - 650 nm, 600 - 700 nm, 650 - 750 nm, 700 - 800 nm, 750 - 850 nm, 800 - 900 nm, 850 - 950 nm, 900 - 1000 nm, 1000 - 10000 nm, 10000 - 20000 nm, 20000 - 30000 nm, 30000 - 40000 nm, 40000 - 50000 nm, 50000 - 60000 nm, 60000 - 70000 nm, 70000 - 80000 nm, 80000 - 90000 nm, or 90000 - 100000 nm. As a non-limiting example, the liposome can be 20 nm in length or diameter.

[0102] Liposomes can be prepared by hydrating the lipids, followed by sizing of the particles and removing of the non encapsulated composition. Liposomes can be prepared by passive loading mechanical dispersion methods and/or active loading methods, or any other suitable method. In some embodiments, liposomes are prepared by thin-film hydration method, micro emulsification, sonication, membrane extrusion, freeze thawed method, ether injection method, ethanol injection method, reverse phase evaporation method, dehydration- rehydration, and/or calcium-induced fusion method. In some embodiments, the liposomes of the disclosure are prepared by passive loading method where the composition is encapsulated by introducing an aqueous phase of a water-soluble components of the composition and/or an organic phase of the lipid-soluble components of the composition, before or at some stage during the preparation of the liposomes.

[0103] In some embodiments, the liposomes can be prepared by the thin-film hydration procedure, where liposomes are prepared by dissolving the phospholipids in organic solvents, such as, dichloromethane, chloroform, ethanol, and/or chloroform-methanol mixture. A thin and homogeneous lipid film can be formed the solvent is evaporated under vacuum at the temperature: about 45-60 °C. Nitrogen gas can be used to completely remove the residual solvent. A solution of distilled water, phosphate buffer, phosphate saline buffer at pH 7.4 and normal saline buffer can be used in hydration step. The time for the hydration process can be about 1 hour to 2 hours at a temperature of about 60-70 °C. In order to obtain full lipid hydration, the liposomal suspension can be left overnight at about 4-6 °C.

[0104] In some embodiments, the liposomes can be prepared by injection methods. For example, the liposomes can be prepared by the ether injection method. This method uses a solution of lipids that can be dissolved in ether and/or diethyl ether/methanol mixture which is slowly injected to an aqueous solution of the composition to be capsulated. The subsequent removal of the organic solvent under reduced pressure can lead to the formation of liposomes. In some embodiments, the liposomes can be prepared by ethanol injection method wherein the ethanolic lipid solution can be rapidly injected to a vast excess of preheated distilled water or TRIS-HCI buffer. The incorporation of the composition in liposomal vesicle can depend on its hydrophilic/hydrophobic character. In some embodiments, the liposomes can be prepared by the sonication method. The sonication method is based on size transformation and involves the subsequent sonication of multilamellar vesicles (MLVs) prepared by thin-film hydration method, using sonic energy usually under an inert atmosphere including nitrogen or argon. The sonication method enables homogenous dispersion of small vesicles using bath type or probe type sonicator with a potential for greater tissue penetration. The bath sonicators can be used for preparation of small unilamellar vesicles (SUVs). In some embodiments, MLVs can be prepared by thin-film hydration method by repeatedly passing the composition and lipids through filters of polycarbonate membranes. The liposomes can be prepared using thin-film hydration method subsequently using an extruder for ten cycles to obtain extruded liposomes with uniform diameters.

[0105] In some embodiments, the liposomes can be prepared by reverse-phase evaporation method. In this method, organic solvents such as diethyl ether/isopropyl ether or mixture of diethyl ether and chloroform (1 :1 v/v) and a mixture of chloroform methanol (2:1 v/v)2 can be used. The organic phase can be immiscible with aqueous phase thus an oil/water emulsion can be created. Phosphate buffer saline or citric-Na2HPO4 buffer can be added to aqueous phase with aim to improve the efficiency of liposome formulations. The formation of liposomes can be allowed by continued rotary evaporation of the organic solvents under vacuum.

[0106] In some embodiments, the liposomes can be prepared by the calcium-induced method. This method is based on adding of calcium to SUVs. The formation of multilamellar vesicles is as result of fusion. The addition of ethylenediaminetetraacetic acid (EDTA) to the preparations can result in the formation of large unilamellar vesicles (LUV) liposomes. The preparation of LUV liposomes can be obtained from acidic phospholipids.

[0107] In some embodiments, the liposomes can be prepared by the freezing and thawing method. The method of freezing and thawing is introduced for increasing the trapped volume of liposomal preparations. The freeze-thaw method is dependent on the ionic strength of the medium and the phospholipid concentration. It influences to a physical disruption of lamellar structure leading to formation of unilamellar vesicles. The unilamellar vesicles are rapidly frozen followed by slow thawing, while the freeze and thawing cycles are repeated.

Gels

[0108] In some embodiments, each dose of the formulation is prepared as liposomes within a gel. In some embodiments, the gel can include an oil and/or a nut butter. [0109] Nut butter in the gel can be peanut butter, almond butter, cashew butter, pistachio butter, hazelnut butter, walnut butter, macadamia nut butter, pecan butter, Brazil nut butter, chestnut butter, pine nut butter, coconut butter, or combinations thereof.

[0110] An oil in the gel can be sunflower oil, flaxseed oil, hemp seed oil, olive oil, sesame oil, grapeseed oil, canola oil (from rapeseed plant), corn oil, peanut oil, palm oil, avocado oil, or combinations thereof.

[0111 ] In some embodiments, an oil in the gel can include at least one medium chain triglyceride (MCT) oil. As used herein, "MCT oil" means a medium chain (6 to 12 carbon) aliphatic fatty acid ester of glycerol. For example, the at least one MCT oil can be present from about 10% to about 69%, or from about 10% to about 40%, or from about 10% to about 30%, or from about 10% to about 20%, or from about 10% to about 15%, or from about 20% to about 60%, or from about 30% to about 50%, or from about 40% to about 45%, based on the total weight of the oil in the gel. For example, the MCT oil can include saturated medium chain fatty acids, for example, a plurality of saturated medium chain fatty acids. In an exemplary embodiment, the MCTs include a mixture of fatty acids having from 6 to 12 carbon atoms. The MCT oil can be derived from a plant such as a fruit or vegetable, for example, a plurality of plants. The MCT oil can contain caprylic acid, an 8-carbon saturated FA. The MCT oil can contain capric acid, a 10-carbon saturated FA. For example, the medium-chain triglycerides can contain a mixture of caprylic acid and capric acid.

[0112] In one embodiment, the formulation comprises a gel of sunflower oil, MCT oil and almond butter. The amount of sunflower oil in the gel can be 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1-10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60-70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more of the gel. The amount of MCT oil in the gel can be 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1-10%, 15- 25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60-70%, 65- 75%, 70-80%, 75-85%, 80-90%, 85-95% or more of the gel. The amount of almond butter in the gel can be 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1 -10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60-70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more of the gel. [0113] The formulations can also include a gelling agent such as, but not limited to agar, gellan gum, carrageenan, pectin, and/or gelatin. Thickening agents can also be added to formulations to prepare gels. Non-limiting examples of thickening agents include far celerane, locust bean gum, guar gum, gum arabic, and/or xanthan gum.

Dosing and Administration

[0114] The determination of an appropriate prescribed dosage of a composition in a formulation in order to achieve a desired biological outcome is within the skill level of an ordinary practitioner of the art.

[0115] In some embodiments, each dose of the compositions of the disclosure can include glutathione in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. In some embodiments, each dose of the compositions of the disclosure can include CoQ10 in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. In some embodiments, each dose of the compositions of the disclosure can include pyrroloquinoline quinone in the amount of the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more. . In some embodiments, each dose of the compositions of the disclosure can include iron binding glycoprotein in the amount of from about 0.1 mg/ml to 1 mg/ml, about 0.5 mg/ml to 5 mg/ml, 1 mg/ml to 10 mg/ml, 5 mg/ml to 50 mg/ml, 10 mg/ml to 100 mg/ml, 50 mg/ml to 150 mg/ml, 100 mg/ml to 200 mg/ml, 150 mg/ml to 250 mg/ml, 200 mg/ml to 300 mg/ml, 250 mg/ml to 350 mg/ml, 300 mg/ml to 400 mg/ml, 350 mg/ml to 450 mg/ml, 400 mg/ml to 500 mg/ml, 450 mg/ml to 550 mg/ml, 500 mg/ml to 600 mg/ml, 550 mg/ml to 650 mg/ml, 600 mg/ml to 700 mg/ml, 650 mg/ml to 750 mg/ml, 700 mg/ml to 800 mg/ml, 850 mg/ml to 950 mg/ml, 900 mg/ml to 1000 mg/ml or more.

[0116] In some embodiments, each dose of the formulation is prepared as gel. When prepared as a gel, each dose of the gel can have a volume of about 0.5 ml, 1 ml, 1.5 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 11 ml, 12 ml, 13 ml, 14 ml, 15 ml, 16 ml, 17 ml, 18 ml, 19 ml, 20 ml or more.

[0117] In some embodiments, each dose of the formulation can include glutathione (about 22 mg/ml); CoQ10 (about 22 mg/ml), pyrroloquinoline quinone (about 2 mg/ml) and/or iron binding glycoprotein e.g., lactoferrin (about 7 mg/ml).

[0118] In some embodiments, each dose of the formulation can include glutathione (about 275 mg); CoQ10 (about 220 mg), and/or pyrroloquinoline quinone (about 22 mg) in 15 ml of a dose of the formulation.

[0119] In some embodiments, each dose is prepared as one or more tablets or capsules. [0120] In some embodiments, the formulation can be taken once daily. In some embodiments, the formulation can be taken twice daily, three times daily, four times daily, or more than four times daily. In one example, the formulation can be taken with meals. In some embodiments, the formulation can be taken immediately following a meal. In some embodiments, the formulation is not mixed with the meal. In some embodiments, the formulation can be taken in the morning, afternoon, evening, and/or night. In one example, the formulation can be taken at the same time every day. In another example, the time the formulation is taken can vary. In one example, the daily intake can be contained in one packaging of the formulation that can be consumed once daily. In another example the daily intake can be split across more than one packaging of the formulation which can be taken together. In another example, the daily intake can be contained in two packages of the formulation and a user can administer two packages of the formulation once daily. In another example, the daily intake can be contained in three packages and a user can administer three packages once daily or administer one package of the formulation three times daily. In another example, the daily intake can be contained in four or more packages of the formulation that can be administered one to four times daily.

[0121 ] The formulations of the disclosure can be administered for 1 day, 1 week, 1 month, 1 year or more. In some embodiments, the formulations can be administered for about 1-30 days, 1 -60 days, 1-90 days, 1-120 days, 1 -150 days or more. In some embodiments, the formulations can be administered for about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 20 days, 30 days, 40 days, 50 days, 60 days, 70 days, 80 days, 90 days, 100 days or more. In some embodiments, the formulations can be administered for 1 week, 2 weeks, 3 weeks, 4 weeks, or more. In some embodiments, the formulations can be administered for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or more.

[0122] Formulations of the present disclosure can be administered in the conventional manner by any route where they are active. Administration can be systemic, parenteral, topical, or oral. As a non-limiting example, administration is oral. In some embodiments, the administration can be, but is not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, buccal, or ocular routes, or intravaginally , by inhalation, by depot injections, or by implants.

Methods of use

[0123] The present disclosure provides various uses and applications for the compositions of the disclosure.

[0124] In some embodiments, compositions of the disclosure can be used by individuals who do not present one or more symptoms associated with a disease, condition or a syndrome. In some embodiments, the compositions of the disclosure can be used to treat or improve symptoms associated with one or more diseases, conditions, or syndrome.

[0125] In some embodiments, the compositions of the disclosure can be used to reduce systemic fatigue, cellular inflammation, and/or reactive oxygen species.

[0126] In some embodiments, the compositions of the disclosure can be used to increase energy potential of cellular systems by increasing mitochondrial biogenesis.

[0127] In some embodiments, the compositions of the disclosure can be used to treat diseases or disorders associated with oxidative stress, such as, but not limited to, chronic fatigue syndrome, cardiovascular diseases, diabetes, cancer, chronic kidney disease, chronic obstructive pulmonary disease (COPD), neurological disease. In some embodiments, the compositions of the disclosure can be used to treat chronic fatigue syndrome and/or major depressive disorder.

[0128] In some embodiments, the compositions of the disclosure can be used to improve lifespan or healthspan of a subject.

[0129] The subject who can benefit from the compositions of the disclosure can be of any age, such as, but not limited to about 1-10 years, 5-15 years, 10-20 years, 15-25 years, 20- 30 years, 25-35 years, 30-40 years, 35-45 years, 40-50 years, 45-55 years, 50-60 years, 55- 65 years, 60-70 years, 65-75 years, 70-80 years or more.

[0130] In some embodiments, compositions of the disclosure can prevent the occurrence of a disease. The term ‘preventing’ or ‘prevention’ refers to a reduction in risk of acquiring or developing a disease or disorder (i.e. , causing at least one of the clinical symptoms of the disease not to develop) in a subject that can be exposed to a disease-causing agent, or predisposed to the disease in advance of disease onset.

Chronic Fatigue Syndrome

[0131 ] In some embodiments, the compositions of the disclosure can be used to treat Chronic Fatigue Syndrome (CFS). CFS can be associated with decreased GSH concentration in the brain and mitochondrial dysfunction can be directly related to CFS and neuroinflammation disorders.

[0132] Chronic fatigue syndrome (CFS) is characterized by profound tiredness, regardless of bed rest. Its symptoms can worsen with physical or mental activity. CFS can happen suddenly and last for years. In some embodiments, CFS is also referred to under other names, such as, epidemic neuromyasthesia, idiopathic chronic fatigue and myalgia syndrome, chronic infectious mononucleosis, myalgic encephalomyelitis, post-viral fatigue syndrome, or fibrositis-fibromyalgia syndrome.

[0133] In some embodiments, the compositions of the disclosure can be used to improve one or more symptoms associated with CFS. In some embodiments, the compositions of the disclosure can improve one or more symptoms by 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1-10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60- 70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more. [0134] The primary symptom of CFS is fatigue, which has a definite onset, and is severe, disabling and affects both physical and mental functioning, and furthermore the fatigue is present for a minimum of 6 months at which it was present for more than 50 % of the time.

[0135] Other symptoms of CFS can include sleep disturbances, which are changes in the duration of sleep and/or quality of sleep. The changes could be hypersomnia or increased sleep, or insomnia or reduced sleep, (which should further be described as either difficulty of getting off to sleep, early wakening, or subjectively disturbed or unrefreshing sleep). In some embodiments, CFS is associated with disability, which, as used herein, can refer to any restriction or lack (resulting from loss of psychological or physiological function) of ability to perform an activity in the manner or within the range considered normal for human being, i.e., things that people cannot do in the areas of occupational, social and leisure activities because of their illness. The disability associated with CFS can be distinguished from impairment of function (e.g., weak legs) and from handicap (e.g., unable to work).

[0136] Mood disturbances such as depressed mood, anhedonia, anxious mood, emotional lability and irritability, can also be associated with CFS. In some embodiments, CFS can be associated with Myalgia, which is pain or aching felt in the muscles. The myalgia can be disproportionate to exertion and can be distinguished from feelings of weakness and pain felt in other areas such as the joints.

[0137] In some embodiments, individuals with CFS can have problems with thinking and memory. Most people with CFS have trouble thinking quickly, remembering things, and paying attention to details. In some embodiments, individuals with CFS can have “brain fog” which is generally used to describe the feeling of not being able to think clearly. In some embodiments, individuals with CFS can have worsening of symptoms while standing or sitting upright. This is called orthostatic intolerance. People with CFS can be lightheaded, dizzy, weak, or faint while standing or sitting up. They can have vision changes like blurring or seeing spots.

[0138] In some embodiments, individuals with CFS can have pain (not associated with an injury), muscle pain and aches, joint pain without swelling or redness, headaches, either new or worsening, tender lymph nodes in the neck or armpits, sore throat, digestive issues, like irritable bowel syndrome, chills and night sweats, allergies and sensitivities to foods, odors, chemicals, light, or noise, muscle weakness, shortness of breath, and/or irregular heartbeat. [0139] Worsening of CFS symptoms after physical or mental activity that would not have caused a problem before illness. This is known as post-exertional malaise (PEM). During PEM, any CFS symptoms can get worse or first appear, including difficulty thinking, problems sleeping, sore throat, headaches, feeling dizzy, or severe tiredness. It may take days, weeks, or longer to recover from a crash. Sometimes patients can be house-bound or even completely bed-bound during crashes.

[0140] In some embodiments, compositions of the disclosure can, within about 30 days, cause a reduction of fatigue, reduction in feeling of tiredness, and improve output and recovery during aerobic exercise.

[0141 ] In some embodiments, compositions of the disclosure can, within about 60 days can result in quicker recall, stronger attention span during tasks, deeper and more restorative sleep, reduction in inflammatory markers within the body in patients with CFS.

[0142] In some embodiments, the compositions of the disclosure can, within about 90 days, increase energy, alertness and activity levels, decrease creatine kinase activity and myoglobin concentration in those with exercise-induced muscle damage

Diabetes

[0143] In some embodiments, the compositions of the disclosure can be used to treat diabetes. The diabetes can be type 1 diabetes, type 2 diabetes or gestational diabetes. Reduced GSH have been associated with insulin resistance associated with type 2 diabetes. Insulin resistance can refer to the reduced or inability of the cells, tissue or organisms to take up glucose in the presence of insulin. In some embodiments, the compositions of the disclosure can reduce insulin resistance by 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1- 10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60- 70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more.

[0144] In some embodiments, the compositions of the disclosure can be used to improve one or more symptoms associated with diabetes including, but not limited to, fatigue, increase urination, thirst, weight loss, hunger, blurry vision, numb or tingling hands and/or feet, increase infections or delayed wound healing.

Major Depressive Disorder

[0145] In some embodiments, the compositions of the disclosure can be used to treat a subject diagnosed with major depressive disorder (MDD). MDD is a mood disorder that can cause a persistent feeling of sadness and loss of interest. Patients with MDD have shown to have pro-inflammatory cytokines (IL-6, IL-8) as well as acute and chronic mitochondrial dysfunction. Compositions of the disclosure can be used improve one or more symptoms associated with MDD, such as, but not limited to feelings of sadness, tearfulness, emptiness or hopelessness, angry outbursts, irritability or frustration, even over small matters, loss of interest or pleasure in most or all normal activities, such as sex, hobbies or sports, sleep disturbances, including insomnia or sleeping too much, tiredness and lack of energy, so even small tasks take extra effort, reduced appetite and weight loss or increased cravings for food and weight gain, anxiety, agitation or restlessness, slowed thinking, speaking or body movements, feelings of worthlessness or guilt, fixating on past failures or self-blame, trouble thinking, concentrating, making decisions and remembering things, frequent or recurrent thoughts of death, suicidal thoughts, suicide attempts or suicide, unexplained physical problems, such as back pain or headaches.

[0146] In some embodiments, the compositions of the disclosure can improve one or more symptoms of MDD symptoms by about 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1- 10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60- 70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more.

Improving healthspan and/or lifespan

[0147] In some embodiments, the compositions of the disclosure provide for increasing or improving healthspan of a subject.

[0148] As used herein, “healthspan” can be defined as the period of life spent free of age- related diseases/conditions. The healthspan of a given subject can therefore refer to the period during which the subject does not have an age-related disease/condition (e.g., an age-related disease described herein). A subject can be considered to not to have a given disease/condition until they are diagnosed with a relevant disease condition.

[0149] A subject having an improved/increased/extended healthspan can remain free of an age-related disease/condition for a longer period as compared to a reference subject. In the context of the present disclosure, a subject administered with the compositions of the disclosure can have an increased healthspan as compared to an equivalent, untreated subject. The increase in healthspan can be by about 5 %, 10 %, 15 %, 20 %, 25 %, 30 %, 35 %, 40 %, 45 %, 50 %, 55 %, 60 %, 65 %, 70 %, 75 %, 80 %, 85 %, 90 %, 95 %, 100%, about 1-10%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 40-50%, 45-55%, 50-60%, 55-65%, 60- 70%, 65-75%, 70-80%, 75-85%, 80-90%, 85-95% or more.

[0150] Alternatively, “healthspan” can also refer to the period of life during which a subject does not exhibit significant age-related deterioration in the function of a tissue, organ or organ system of the subject. The deterioration in function can arise as a consequence of an age- related disease/condition. A subject displaying deterioration in a function of a given tissue/organ/organ system can display a level of the relevant function which is less than 1 times, e.g. one of <0.99 times, <0.95 times, <0.9 times, <0.85 times, <0.8 times, <0.75 times, <0.7 times, <0.65 times, <0.6 times, <0.55 times, <0.5 times, <0.45 times, <0.4 times, <0.35 times, <0.3 times, <0.25 times, <0.2 times, <0.15 times or <0.1 times the level observed in comparable subjects not experiencing deterioration in the relevant function.

[0151 ] In some embodiments, the compositions can also be used to extend lifespan. As used herein, lifespan can be defined as the length of time for which a person lives. The compositions of the disclosure can increase lifespan by about 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 15 years, 20 years, 25 years, 30 years, 35 years, 40 years, 45 years, 50 years, about 1 -2 months, 2-3 months, 3-4 months, 4-5 months, 5-6 months, 6-7 months, 7-8 months, 8-9 months, 9-10 months, 10-11 months, 1 -2 years, 2-3 years, 3-4 years, 4-5 years, 5-6 years, 6-7 years, 7-8 years, 8-9 years, 9-10 years, 10-20 years, 15-25 years, 20-30 years, 25-35 years, 30-40 years, 35-45 years, 40-50 years or more.

[0152] The compositions and methods are more particularly described below, and the Examples set forth herein are intended as illustrative only, as numerous modifications and variations therein will be apparent to those skilled in the art. The terms used in the specification generally have their ordinary meanings in the art, within the context of the compositions and methods described herein, and in the specific context where each term is used. Some terms have been more specifically defined herein to provide additional guidance to the practitioner regarding the description of the compositions and methods.

[0153] As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items. As used in the description herein and throughout the claims that follow, the meaning of “a”, “an”, and “the” includes plural reference as well as the singular reference unless the context clearly dictates otherwise. The term “about” in association with a numerical value means that the value varies up or down by 5%. For example, for a value of about 100, means 95 to 105 (or any value between 95 and 105).

[0154] All patents, patent applications, and other scientific or technical writings referred to anywhere herein are incorporated by reference herein in their entirety. The embodiments illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that are specifically or not specifically disclosed herein. Thus, for example, in each instance herein any of the terms "comprising," "consisting essentially of," and "consisting of" can be replaced with either of the other two terms, while retaining their ordinary meanings. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claims. Thus, it should be understood that although the present methods and compositions have been specifically disclosed by embodiments and optional features, modifications and variations of the concepts herein disclosed can be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of the compositions and methods as defined by the description and the appended claims.

[0155] Any single term, single element, single phrase, group of terms, group of phrases, or group of elements described herein can each be specifically excluded from the claims.

[0156] Whenever a range is given in the specification, for example, a temperature range, a time range, a composition, or concentration range, all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the disclosure. It will be understood that any subranges or individual values in a range or subrange that are included in the description herein can be excluded from the aspects herein. It will be understood that any elements or steps that are included in the description herein can be excluded from the claimed compositions or methods

[0157] In addition, where features or aspects of the compositions and methods are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the compositions and methods are also thereby described in terms of any individual member or subgroup of members of the Markush group or other group. [0158] The following are provided for exemplification purposes only and are not intended to limit the scope of the embodiments described in broad terms above.

EXAMPLES

Example 1: In vivo studies in Caenorhabditis elegans

C. elegans (worm) maintenance and media

[0159] To prevent chemical modification or metabolism of the test article by the food bacteria, animals are fed on a lawn of inactivated E. coli, strain OP50. Cultures of OP50 are inactivated by exposure to 0.25% paraformaldehyde for 1 hour followed by 5 washes in M9. Bacteria are dispersed by passing through a 5pM filter during the wash steps. The quantity and distribution of food bacteria are calibrated to ensure adequate access to food for the duration of assay while maintaining visibility of the animals.

Reactive oxygen species (ROS) assay

[0160] A large population of wild-type C. elegans are synchronized and grown to the L4 juvenile stage. Liquid culture medium is prepared as follows: S medium, 50 uM 5- Fluorodeoxyuridine (FUdR), 200 pg/ml Streptomycin, and filtered OP50. The L4 animals are resuspended in liquid culture at about 2 animals/pl. 90pl of the animals in the liquid culture media are added to each well. Compositions of the disclosure are extracted into ethanol, water and/or DMSO and 10 pl are added to each well. Micro plates are sealed and incubated at 20°C overnight on shaker. The next day, activity is measured on the WMicrotracker (PhylumTech) for 2 hours and a basal measurement is obtained. 11 pl of Paraquat (1 OOmM) is pipetted to each well for a final concentration of 10mM. The plate is resealed and activity is recorded on the WMicrotracker for 48 hours. The activity is grouped in blocks of 40 minutes and percent activity is calculated by normalizing to the basal activity measurements.

[0161 ] The ROS assay is expected to show that pre-treatment with the compositions of the disclosure can provide improved protection against reactive oxygen species (ROS) than the positive control, paraquat.

Lifespan assay

[0162] The lifespan assay is initiated by expanding all groups (group treated with compositions of the disclosure and control group) to more than 1000 animals, then synchronizing by bleaching and allowing larval animals to hatch and arrest. To eliminate the effect of bacterial metabolism and growth on lifespan, synchronized animals are only exposed to dead food bacteria. To suppress progeny, animals are transferred to media containing 5-Fluorodeoxyuridine (FUdR) within 54-60 hours post-plating. Animals are inspected 24 and 48 hours after this transfer to confirm infertility. Finally, the animals are inspected for general health and morphology before transferring to scanner plates. The scanner plates are incubated for an additional 2 days and inspected again before loading onto scanners.

[0163] To understand the physiological impact of either genetic variation or pharmacological treatment on aging, quantitative models are used. The analysis of lifespan data is grounded in the study of two mathematical functions: the survival curve and the hazard function. The survival curve describes the fraction of a tested population that remains alive over time. The hazard function is related to the survival curve and provides an intuitive measure of the risk of death. This function describes the probability that a typical individual who is currently alive will soon die, providing a clear visualization of the way a treatment can change patterns in mortality. To obtain high-resolution lifespan data and eliminate confounding factors such as worm handling and operator bias, lifespan data is collected using an Automated Lifespan Machine (ALM) which is adapted from Stroustrup, N. et al., Nat Methods 10, 665-670 (2013) (the contents of which are herein incorporated by reference in its entirety). Three biological replicates are derived from synchronizing three independently maintained lines of N2 animals. Images of the animals are then collected for the next 35 days with no interruption or manipulations.

[0164] The lifespan of the animals treated with the compositions of the disclosure is expected to increase significantly compared to untreated controls.

Survival Analysis

[0165] Time of death calls exported from the ALM software is analyzed and plotted using the Lifelines software package developed by Cam Davidson-Pilon et al., CamDavidsonPilon/lifelines: vO.25.9 (https://doi.org/10.5281/zenodo.4505728; the contents of which are herein incorporated by reference in its entirety). Additional analysis is performed using the OASIS2 analysis software. A standard Mantel-Cox log-rank test is used to compare the curves globally over the course of the lifespan assigning equal weight to each timepoint, whereas the Wilcoxon-Breslow-Gehan weights each death by the number of subjects at risk, assigning greater weight to earlier deaths. Each of these tests is expected to show a significant increase in survival upon treatment with the compositions of the disclosure compared to the control. [0166] The survival rate is expected to increase significantly in animals treated with the compositions of the disclosure compared to untreated controls.

Movement and healthspan analysis

[0167] Worm movement is tracked from the images acquired by the ALM during the lifespan assay. Worm size and movement features are extracted and analyzed using custom software. Worm Activity serves as a proxy for animal health. Changes in spatial distribution of the animals between time points is used to derive gross movement for the population over time. Foreground motion is calculated using an approach called difference based spatial temporal entropy image (DSTEI) (see Ma, Yu-Fei et al., In IEEE International Conference on Multimedia and Expo, 2001. ICME 2001., 265-68; the contents of which are herein incorporated by reference in its entirety). Worm morphology is measured from the worm contours detected in the images. In the process of aging, animals become shorter and stouter over time and their shape is an indicator of their overall health and biological age. The length is calculated from the central spline fitted to the worm contour and width is measured from each worm’s widest point. The animals’ posture also changes with age as they lose the ability to maintain an elongated position. Average Circularity measures how close the shape and posture come to being enclosed by a circle. Each of these measures are obtained by averaging data for all active animals detected on a plate, then averaging across different replicate plates of the same condition. All measurements are based on animals that are still alive and moving at the time of quantification. All measures are initiated animals are placed on the scanner at day 4 of adulthood.

[0168] Animals treated with the compositions of the disclosure are expected to show higher levels of activity sustained throughout the lifespan, with the higher dosage showing higher activity.

[0169] Animals treated with the compositions of the disclosure are expected to experience an increase in healthspan.

Whole transcriptome analysis

[0170] More than 150-day 1 adult animals per replicate are harvested, cleaned by filtration, and frozen at -80° in Trizol. To extract RNA, samples are thawed, vigorously vortexed, and processed using the Direct-zol RNA Miniprep Kit (Zymo Research).

[0171 ] The total RNA is enriched for poly-mRNA using oligo(dT) paramagnetic beads. DNA libraries are then constructed from this input mRNA using the NEBNext UltraTM II RNA Library Prep Kit to create a ready-to-sequence dsDNA library that retains the strand-specific information in the original mRNA. These libraries are then further tested by the Qubit for concentration and the Agilent 2100 for library size distribution and quality. In order to properly pool the libraries and load them onto sequencing lanes to ensure the correct number of reads per sample, an even more precise quantification of the library is done via qPCR, and the samples are loaded onto the NovaSeq 6000 platform for a paired-end sequencing run of 150 bp for each end (PE150). The loading concentrations are designed to obtain at least 6.0 Gb (which is the number of billion bases of raw data, determined by the number of reads multiplied by the length of each read).

[0172] Differentially expressed genes (DEGs) are mapped to known physiological pathways and then examined for coherent linkages between pathways related to longevity. These pathways are drawn from WormBase, KEGG, and other published databases and literature. The transcriptomic analysis is expected to show that treatment with compositions of the disclosure can result in changes in expression of genes involved with insulin response, energy metabolism, and canonical longevity pathways.