Preparations for reducing or preventing oxidative stress damage

Abstract

A composition comprising: a flavonoid, for example, in an amount of about 0.1 g to about 1.5 g; and one or more of: ascorbic acid, ascorbate, or a combination thereof, for example, in an amount of about 0.2 g to about 2.0 g; N-acetylcysteine, for example, in an amount of about 0.10 g to about 1.2 g; α-lipoic acid, for example, in an amount of about 0.05 g to about 0.60 g; and at least one carotenoid, for example, in an amount of about 1 mg to about 50 mg. The composition can be used to reduce or prevent nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in a subject caused by oxidative stress.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 926,953, filed on October 28, 2019, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to a combination of compounds that can be used to reduce or prevent nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in a subject caused by oxidative stress. Background Art

[0004] The following paragraphs are not an admission that anything discussed therein is prior art or part of the general knowledge of a person skilled in the art.

[0005] Oxidative stress occurs when a biological system is unable to detoxify reactive oxygen species or repair damage caused by them at a sufficiently fast rate compared to the rate at which they are produced or the rate at which they are damaged. Elevated levels of reactive oxygen species, such as peroxides and free radicals, can damage proteins, lipids, and DNA. Summary of the Invention

[0006] The following description is intended to introduce the reader to this specification but does not limit any inventions. One or more inventions may reside in any combination or subcombination of the device elements or method steps described below or elsewhere in this document. The inventors do not waive or disclaim their rights to any one or more inventions disclosed in this specification solely by failing to recite such other one or more inventions in the claims.

[0007] Oxidative stress can result from acute exposure to ionizing radiation, such as doses greater than 0.05 mSv over a period of less than 5 hours. Ionizing radiation can come from surgical procedures using X-ray guidance, computed tomography (CT), radioactive tracers, or other sources of ionizing radiation; from medical imaging procedures such as X-rays, CT, or dental X-rays; from medical procedures using radioactive materials, contrast agents, or radioisotopes; or from environmental exposures such as air travel, industrial X-ray use, or proximity to radiation sources (e.g., nuclear power plants, nuclear waste storage sites, or accidental or intentional releases of radioactive materials).

[0008] Oxidative stress can be caused by smoking or vaping, for example, by inhaling or vaping tobacco or cannabis products. While not wishing to be bound by theory, it is believed that smoking or vaping increases oxidative stress through direct damage from reactive oxygen species and / or other free radical species (i.e., the inflammatory response caused by smoking or vaping, or both).

[0009] Elevated levels of reactive oxygen species, such as those produced during oxidative stress, can damage cells. Cumulative cellular damage is associated with an increased risk of various diseases and conditions, such as cancer, cataracts, and vascular and cardiovascular disease. Even small reductions in an individual's average oxidative stress levels can be beneficial because chronic, low-grade inflammation can overwhelm damage-repair pathways and the signals that initiate such repair processes.

[0010] Because chronic and / or cumulative exposure to ionizing radiation, cigarette smoking, or vaping can result in protein, lipid, and / or DNA damage associated with oxidative stress, it is desirable to provide compositions of compounds that reduce oxidative stress when administered to a subject.

[0011] The present invention provides a composition comprising: a flavonoid and one or more of: ascorbic acid or ascorbate; N-acetylcysteine; alpha-lipoic acid; at least one carotenoid; and folic acid, folate, or methylfolate. In a specific example, the present invention provides a composition comprising a flavonoid; ascorbic acid or ascorbate; N-acetylcysteine; alpha-lipoic acid; optionally at least one carotenoid; and optionally folic acid, folate, or methylfolate. Some compositions according to the present invention can be used to reduce or prevent nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in a subject caused by oxidative stress. Some compositions according to the present invention can be used to help repair nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in a subject. DETAILED DESCRIPTION

[0012] In general, the present invention provides a composition comprising a flavonoid and one or more of: ascorbic acid or an ascorbic acid subject; N-acetylcysteine; alpha-lipoic acid; at least one carotenoid; and folic acid, a folic acid subject, or a methylfolate subject.

[0013] It should be understood that in the context of the present invention, a "combination" is not limited to a preparation in which all components of the composition are physically combined together. On the contrary, a "combination" according to the present invention refers to components that are physically separated but intended for administration to a subject within about a one-hour window.

[0014] For example, a composition according to the present invention can be formulated so that all compounds of the composition can be administered to a subject together. A specific example of such a formulation is a water dispersible powder.

[0015] In another example, the composition according to the present invention can physically separate the water-soluble components from the water-insoluble and water-unstable components. The water-soluble components can be formulated in a water-based beverage, while the water-insoluble and water-unstable components can be formulated in a non-aqueous edible gel or a non-aqueous drinkable liquid.

[0016] The flavonoids used in the compositions according to the present invention may have antioxidant properties. In some instances, the flavonoid is a flavanol, such as quercetin or a quercetin derivative. In other instances, the flavonoid is a flavanonol, such as dihydroquercetin or a dihydroquercetin derivative. The compositions according to the present invention may comprise a mixture of flavonoids.

[0017] Flavonoid "derivatives," such as quercetin derivatives or dihydroquercetin derivatives, can be glycosylated flavonoids or prodrug flavonoid analogs. Examples of glycosylated quercetin include quercetin-3-O-paltoside, quercetin-3-O-glucoside, quercetin-3-O-rutinoside, quercetin-3-O-galactoside, and quercetin-3-O-rhamnoside. Examples of glycosylated dihydroquercetin include: dihydroquercetin-3-O-rhamnoside; dihydroquercetin-3-O-glucoside; (-)-2,3-trans-dihydroquercetin-3'-O-β-D-glucopyranoside; (2S,3S)-(-)-dihydroquercetin-3-O-β-D-glucopyranoside; (2R,3R)-dihydroquercetin-3'-O-β-D-pyranoglucoside; dihydroquercetin-4'-O-β-glucopyranoside; (2R,3R)-dihydroquercetin-3-O-arabinoside; and (2S,3S)-dihydroquercetin-3-O-arabinoside. Examples of prodrug flavonoid analogs are ester-modified quercetin or glycosylated quercetin. The ester modification can be removed by esterases or hydrolysis to reveal quercetin or glycosylated quercetin.

[0018] Quercetin and / or quercetin derivatives can be extracted or isolated from apple peel, onion or Sophora japonica seeds or flowers. Exemplary methods are discussed in U.S. Patent No. 9,101,649, which is incorporated herein by reference. In one example, quercetin and / or quercetin derivatives can be extracted from apple peel (e.g., from dried apple peel powder) using a food-grade solvent (e.g., ethanol). The apple peel can be ultrasonically treated during the extraction process. The extracted quercetin and / or quercetin derivatives can be separated from the remaining solids and can be optionally concentrated, dried and / or frozen. The extracted quercetin and / or quercetin derivatives can be purified, for example, by column chromatography.

[0019] In the composition according to the present invention: the mass / mass ratio of flavonoids to ascorbic acid or ascorbate may be from about 1:10 to about 5:4; the mass / mass ratio of flavonoids to N-acetylcysteine ​​may be from about 1:5 to about 5:1; the mass / mass ratio of flavonoids to α-lipoic acid may be from about 1:5 to about 5:1; the mass / mass ratio of flavonoids to at least one carotenoid may be from about 200:1 to about 10:1; the mass / mass ratio of flavonoids to folic acid, folate or methylfolate may be from about 10000:1 to about 100:1; or any combination thereof.

[0020] In the composition according to the present invention: the amount of flavonoids may be from about 0.1 g to about 1.5 g; the amount of ascorbic acid or ascorbate may be from about 0.2 g to about 2.0 g; the amount of N-acetylcysteine ​​may be from about 0.10 g to about 1.2 g; the amount of α-lipoic acid may be from about 0.05 g to about 0.60 g; the amount of at least one carotenoid may be from about 10 mg to about 50 mg; the amount of folic acid, folate or methylfolate may be from about 100 μg to about 400 μg; or any combination thereof.

[0021] A specific example of a composition according to the present invention comprises: a flavonoid; ascorbic acid or ascorbate; N-acetylcysteine; and alpha-lipoic acid.

[0022] The composition according to the present invention may comprise at least one carotenoid, which may be a carotene compound, such as beta-carotene; a xanthophyll compound, such as lutein, astaxanthin, zeaxanthin, or a combination thereof; or a combination thereof. The composition comprising at least one carotenoid may reduce or prevent nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in epithelial cells (e.g., vascular epithelial cells, ocular epithelial cells, or both); connective tissue cells; muscle tissue cells; nerve cells; or any combination thereof. DNA damage may comprise double-strand breaks, single-strand breaks, oxidative damage (e.g., 8-OH-2-deoxyguanosine), or a combination thereof.

[0023] In the context of the present invention, reducing or preventing nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof is understood to mean that, after an oxidative stress event, the subject population administered the composition according to the present invention has statistically less nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or a combination thereof in at least one tissue or cell type compared to an otherwise statistically identical subject population not administered the composition. For example, reducing or preventing lipid peroxidation is understood to mean that, after an oxidative stress event, the subject population administered the composition according to the present invention has statistically less lipid peroxidation in skin cells compared to primary fibroblasts in an otherwise statistically identical subject population not administered the composition.

[0024] In the context of the present invention, nuclear DNA damage was assessed by counting the total number of double-stranded DNA breaks per cell in peripheral blood mononuclear cells (PBMCs) using a three-dimensional microscope and fluorescently labeled γ-H2AX protein. γ-H2AX protein is a phospho-isoform of histone H2A that is phosphorylated during the detection of double-stranded DNA breaks to promote the aggregation of repair proteins. To quantify nuclear DNA damage, γ-H2AX was fluorescently labeled and the total number of double-stranded breaks per cell was counted by fluorescence microscopy. γ-H2AX was fluorescently labeled using the Abcam γH2A.X staining kit (ab242296) according to the manufacturer's protocol.

[0025] In the context of the present invention, long-range PCR (LR-PCR) and digital PCR were used to assess mitochondrial DNA damage. The LR-PCR method uses extracted mitochondrial DNA (mtDNA) that is amplified using a high-fidelity DNA polymerase-mediated long-range polymerase chain reaction (PCR) method, as discussed in Gianni P. et al. Exp. Gerontol. 2004 Sep; 39(9): 1391-400. Mitochondrial DNA damage is quantified as the amount of full-length amplifiable mitochondrial DNA and the abundance of mitochondrial DNA deletions.

[0026] In the context of the present invention, total DNA damage was measured using 8-hydroxy-2'-deoxyguanosine (8-OHdG) in cell lysates using the 8-Oxo Detection Elisa kit (Caymanchemical 589320) according to the manufacturer's protocol.

[0027] Lipid peroxidation refers to the oxidative degradation of lipids to form reactive aldehydes such as malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). MDA and 4-HNE are commonly used as markers of lipid peroxidation. In the context of the present invention, lipid peroxidation is assessed by fluorescent Western-blot analysis to quantify the amount of malondialdehyde (MDA) or 4-hydroxynonenal (4-HNE) in a sample. Lipid peroxidation is measured in cell lysates by Western blotting using anti-4-HNE antibodies, as discussed in Kitaoka, Y. et al., Mol Genet Metab. 2013 Nov; 110(3): 297-302. It should be understood that as long as the amount of MDA, the amount of 4-HNE, or the combined amount of MDA and 4-HNE is statistically lower in a population of subjects administered a composition according to the present invention compared to a statistically identical population of subjects not administered the composition, mitigation or prevention of lipid peroxidation can be achieved.

[0028] Protein carbonylation is a form of protein oxidation in which reactive ketones and / or aldehydes are formed. In the context of the present invention, protein carbonylation is assessed by fluorescence analysis to quantify the amount of ketones and / or aldehydes in a sample. The ketones and / or aldehydes are first reacted with 2,4-dinitrophenylhydrazine (DNPH) to form hydrazones, which are then quantified using Western blotting as discussed in Kitaoka, Y. et al., Mol Genet Metab, 2013 Nov; 110(3): 297-302.

[0029] The oxidative stress event can be, for example, acute exposure to ionizing radiation, or repeated exposure to subacute levels of ionizing radiation. The tissue or cell type can be bone marrow, skeletal muscle (eg, quadriceps), skin tissue, lung tissue, or heart tissue.

[0030] The composition according to the present invention may comprise folic acid, folate or methylfolate. The composition comprising folic acid, folate or methylfolate can be used to reduce or prevent nuclear DNA damage caused by oxidative stress in a subject suffering from stroke and / or heart attack.

[0031] Compositions according to the present invention may include an iodide, such as potassium iodide. The mass / mass ratio of flavonoid to iodide may be from about 10:1 to about 1:1. The amount of iodide may be from about 30 mg to about 160 mg. Compositions comprising iodide may be used to reduce or prevent absorption of radioactive iodine by a subject, for example, by the subject's thyroid gland.

[0032] The composition according to the present invention may comprise antioxidant glycosylated or non-glycosylated isoflavones, such as genistein or daidzein. The composition comprising antioxidant glycosylated or non-glycosylated isoflavones may reduce or prevent nuclear DNA damage caused by oxidative stress in skin inflammation induced by exposure to UV radiation (e.g., solar UV radiation).

[0033] The composition according to the present invention may comprise an antioxidant phytocannabinoid, such as cannabidiol (CBD). Compositions comprising antioxidant phytocannabinoids can alleviate or prevent nuclear DNA damage caused by oxidative stress of skin inflammation induced by exposure to UV radiation (e.g., solar UV radiation). Compositions comprising CBD can be formulated for transdermal and oral administration, wherein CBD and flavonoids are formulated for transdermal administration and the remaining ingredients are formulated for oral administration. Transdermal formulations containing CBD can provide analgesic effects.

[0034] Compositions according to the present invention may include radioactive imaging contrast agents, such as barium sulfate or gadolinium-based or iodine-based contrast agents. Radioactive imaging contrast agents may affect the DNA breakage of patients, for example, when patients receive chemotherapy and X-ray treatment. Compositions comprising radioactive imaging contrast agents can reduce or prevent nuclear DNA damage caused by medical imaging procedures. For example, a composition of barium sulfate, flavonoids (such as quercetin), ascorbic acid or ascorbate, N-acetylcysteine ​​and α-lipoic acid can be formulated for oral administration. Such preparations can be given to patients before gastrointestinal (GI) fluoroscopy or before CT scanning. In another example, gadolinium-based contrast agents can be formulated for intravenous administration, and mixtures of flavonoids (such as quercetin, ascorbic acid or ascorbate, N-acetylcysteine ​​and α-lipoic acid) can be formulated for oral administration.

[0035] The composition according to the present invention can be formulated for oral administration or transdermal administration, or a combination of oral and transdermal administration.

[0036] When exposed to water for a long time, such as several days, weeks or months, some flavonoids included in the composition according to the present invention are unstable. The preparation comprising such water-unstable flavonoids is essentially anhydrous, although the essentially anhydrous preparation can be mixed with water before administration (e.g., up to one hour before administration). In the context of the present invention, "essentially anhydrous" is understood to mean that the flavonoids in the preparation degrade no more than 10 mol% after one week due to the presence of water in the preparation. Some preparations can include a measurable amount of water, but are still considered to be essentially anhydrous. For example, if a carbohydrate-based gel forms enough hydrogen bonds with the water present in the preparation to prevent water from degrading more than 10 mol% of the flavonoids (as measured after one week), then the gel can be considered to be essentially anhydrous.

[0037] Formulations for oral administration can be water-dispersible powders, tablets, capsules, edible gels, potable liquids, or any combination thereof. Water-dispersible powders can contain, for example, microcrystalline cellulose (MCC), calcium chloride, or both. Microcrystalline cellulose aids in the dispersion of water-insoluble ingredients (such as water-insoluble flavonoids) in the potable liquid. Tablets can contain MCC, calcium chloride, magnesium stearate, sodium carbonate, bicarbonate, and / or citric acid.

[0038] In one example, a formulation for oral administration can be a combination of: (i) a water-based edible gel or drinkable liquid comprising one or more water-soluble ingredients (e.g., ascorbic acid or ascorbate, N-acetylcysteine ​​and / or folic acid, folate or methylfolate), and (ii) a substantially anhydrous powder, anhydrous edible gel or anhydrous drinkable liquid comprising at least a flavonoid, and optionally α-lipoic acid and / or a carotenoid.

[0039] In a specific example, a drinkable liquid comprising ascorbic acid or an ascorbate salt, N-acetylcysteine, and folic acid, folate, or methylfolate can be provided in a packet containing a substantially anhydrous powder or gel comprising quercetin, alpha-lipoic acid, and beta-carotene. The powder or gel can be mixed with the drinkable liquid and the resulting mixture can be ingested.

[0040] The compositions according to the invention can be formulated in dosages suitable for a particular risk profile associated with a particular exposure to ionizing radiation. For example, a composition of a compound can be formulated in dosages suitable for a patient who will be exposed to a particular dose of X-rays.

[0041] Alternatively, the composition of the compound can be formulated into a dosage that is combined with additional doses based on the risk profile associated with exposure to ionizing radiation. For example, the composition of the compound can be formulated into a dosage in which administration of a single dose is appropriate for a person with a low risk profile, while administration of multiple doses (e.g., two or three doses) is appropriate for a person with a high risk profile.

[0042] In the context of the present invention, a person with a "low risk" profile is one who is likely to be exposed to about 0.05 mSv to about 5 mSv in a 5-hour period. For example, a person with a low risk profile may be a hospital employee, such as a doctor or nurse, who is exposed to low levels of ionizing radiation every day or almost every day while working. A person with a low risk profile may be given a prophylactic dose of the composition on the days they go to work. In another example, a person with a low risk profile may be a traveler, pilot, or flight attendant who flies for at least 6 hours. A dose of the composition may be administered to such a person with a low risk profile starting 0 to about 4 hours before the start of the flight and then every about 6 to about 12 hours.

[0043] In the context of the present invention, a person with a "high risk" profile is one who is likely to be exposed to more than 5 mSv within 5 hours. For example, a person with a high risk profile may be one who is exposed to high levels of ionizing radiation, for example due to accidental exposure to radioactive material.

[0044] In some examples, a composition according to the present invention comprises about 2.0 g of ascorbic acid or an ascorbate salt, about 1.2 g of N-acetylcysteine, about 0.6 g of alpha-lipoic acid, about 0.5 g of quercetin, about 30 mg of beta-carotene, and about 400 μg of folic acid, folate, or methylfolate. Such a composition can be formulated for oral administration to a subject, for example, in a water-dispersible powder that can be mixed with a water-based beverage (e.g., a barium sulfate suspension) to provide a drinkable formulation. The subject can ingest the formulation about 2 hours before a planned exposure to ionizing radiation (e.g., a medical imaging procedure).

[0045] In some examples, a composition according to the present invention comprises about 0.8 g of ascorbic acid or ascorbate, about 0.6 g of N-acetylcysteine, about 0.5 to about 0.6 g of alpha-lipoic acid, about 0.5 g of quercetin, about 15 mg of beta-carotene, and about 200 μg to about 400 μg of folic acid, folate, or methylfolate. This combination can be formulated for oral or transdermal administration to subjects with a "low-risk" profile. For example, the composition can be administered to a subject about one or two hours before going to work or flying.

[0046] In some examples, a composition according to the present invention comprises about 0.8 g ascorbic acid or ascorbate, about 0.6 g N-acetylcysteine, about 0.6 g alpha-lipoic acid, about 0.5 g quercetin, about 15 mg beta-carotene, about 400 μg folic acid, folate, or methylfolate, and about 130 mg potassium iodide. Such a composition can be formulated for oral or transdermal administration to a subject who may have been exposed to depleted or enriched uranium. For example, the subject can be an individual involved in the cleanup of a radioactive industrial accident; or an individual exposed to weapons containing or releasing depleted or enriched uranium.

[0047] In some examples, a composition according to the present invention comprises about 1.0 g ascorbic acid or an ascorbate salt, about 0.6 g N-acetylcysteine, about 0.3 g alpha-lipoic acid, about 0.25 g quercetin, about 15 mg beta-carotene, and about 200 μg folic acid, folate, or methylfolate. Such a composition can be formulated for oral administration, for example, in a water-dispersible powder. A single dose of the composition can be suitable for daily or near-daily administration in a person with a low-risk profile, while a two- or three-dose composition can be suitable for one-time administration or for short-term (e.g., about 1 week to about 6 months) daily administration in a person with a high-risk profile.

[0048] In some examples, a composition according to the present invention comprises about 2.0 g of ascorbic acid or ascorbate, about 1.2 g of N-acetylcysteine, about 0.6 g of alpha-lipoic acid, about 0.5 g of quercetin, about 30 mg of beta-carotene, and 400 μg of folic acid, folate, or methylfolate. Such a composition can be formulated for oral and transdermal administration to subjects who have been or are expected to be exposed to UV radiation. The portion of the composition formulated for oral administration can comprise ascorbic acid or ascorbate, N-acetylcysteine, alpha-lipoic acid, beta-carotene, and folic acid, folate, or methylfolate. The portion of the composition formulated for transdermal administration, for example in a cream or ointment, can comprise quercetin and optionally cannabidiol.

[0049] The composition according to the present invention may additionally comprise coenzyme Q10 (CoQ10), for example, in an amount of about 100 mg to about 500 mg. The flavonoids and Coenzyme Q10 may be present in a mass / mass ratio of about 5:1 to about 1:5, for example about 1:1.

[0050] The composition according to the present invention may additionally comprise tocopherol, such as vitamin E, in an amount of about 100 mg to about 500 mg. The flavonoid and tocopherol (such as vitamin E) may be present in a mass / mass ratio of about 5:1 to about 1:5, such as about 1:1.

[0051] The composition according to the present invention may additionally comprise selenium, selenomethionine, selenocysteine, selenate, or any combination thereof, for example, in an amount of about 20 μg to about 500 μg. The flavonoids and selenium, selenomethionine, selenocysteine, selenate, or any combination thereof may be present in a mass / mass ratio of about 10,000:1 to about 100:1, for example, about 2,000:1.

[0052] The composition according to the present invention may additionally comprise vitamin B, such as vitamin B12, in an amount of about 10 μg to about 250 μg. The flavonoids and vitamin B (such as vitamin B12) may be present in a mass / mass ratio of about 20,000:1 to about 100:1 (e.g., about 4,000:1).

[0053] Compositions according to the present invention may comprise quercetin; coenzyme Q10; alpha-lipoic acid; vitamin E; and ascorbic acid, ascorbate, or both ascorbic acid and ascorbate; and optionally folic acid, folate, methylfolate, vitamin B12, or any combination thereof; optionally beta-carotene, lutein, astaxanthin, zeaxanthin, or any combination thereof; optionally, selenium, selenomethionine, selenocysteine, selenate, or any combination thereof; and optionally an iodide, such as potassium iodide.

[0054] A particular composition according to the invention comprises: about 200 mg quercetin; about 200 mg coenzyme Q10; about 200 mg alpha-lipoic acid; about 200 mg vitamin E; about 200 mg ascorbic acid or ascorbate, or a combination thereof; about 4 mg astaxanthin; about 2.5 mg zeaxanthin; about 400 μg folic acid, folate, methylfolate, or a combination thereof; about 100 μg selenium; and about 50 μg vitamin B12.

[0055] The composition according to the present invention can be formulated for oral administration, transdermal administration, or a combination thereof. The formulation for oral administration can be: powder, tablet, capsule, edible gel, drinkable liquid or any combination thereof.

[0056] Formulations for oral administration may be a combination of a water-based edible gel or drinkable liquid comprising one or more water-soluble ingredients (e.g., ascorbic acid or an ascorbate salt, N-acetylcysteine ​​and / or folic acid, folate or methylfolate), and a substantially anhydrous powder, substantially anhydrous edible gel or substantially anhydrous drinkable liquid comprising the flavonoid and optionally alpha-lipoic acid and / or carotenoid.

[0057] Formulations for oral administration may be water dispersible powders, effervescent tablets, or formulated as drinking liquids to be mixed with water-based beverages. Formulations for transdermal administration may be topical creams or ointments, or transdermal patches.

[0058] A formulation for combined oral and transdermal administration may comprise (i) an oral formulation comprising one or more water-soluble ingredients (e.g., ascorbic acid or ascorbate, N-acetylcysteine ​​and / or folic acid, folate or methylfolate), and (ii) a transdermal formulation comprising a flavonoid and optionally α-lipoic acid and / or a carotenoid.

[0059] The composition may be formulated as a powder, one or more tablets, or one or more capsules for oral administration.

[0060] The composition can be formulated as 2 to 5 capsules for oral administration; for example, where the composition is formulated for oral administration as three capsules, each capsule comprises: about 67 mg quercetin; about 67 mg coenzyme Q10; about 67 mg alpha-lipoic acid; about 67 mg vitamin E; about 67 mg ascorbic acid or ascorbate, or a combination thereof; about 1.3 mg astaxanthin; about 0.83 mg zeaxanthin; about 133 μg folic acid, folate, methylfolate, or a combination thereof; about 33 μg selenium; and about 17 μg vitamin B12.

[0061] Any composition according to the present invention can be used to reduce or prevent nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation, or any combination thereof in a subject caused by oxidative stress.

[0062] Nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, or protein carbonylation is present in epithelial cells of the subject, such as cardiovascular epithelial cells or corneal epithelial cells; connective tissue cells of the subject; muscle tissue cells of the subject; neural cells of the subject; or any combination thereof.

[0063] The epithelial cells may be cardiovascular epithelial cells or corneal epithelial cells.

[0064] Oxidative stress can result from exposure to ionizing radiation, such as from medical imaging procedures, the release of nuclear material, or air travel, or from skin inflammation caused by exposure to ultraviolet radiation.

[0065] Oxidative stress can result from high exposure to reactive oxygen species from smoking or vaping tobacco or marijuana products.

[0066] The composition according to the present invention can be used in a method for reducing or preventing nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, protein carbonylation or any combination thereof in a subject caused by oxidative stress. The method comprises administering the composition to the subject.

[0067] Nuclear DNA damage, mitochondrial DNA damage, lipid peroxidation, or protein carbonylation is present in epithelial cells of the subject; connective tissue cells of the subject; muscle tissue cells of the subject; neural cells of the subject; or any combination thereof.

[0068] The epithelial cells may be cardiovascular epithelial cells or corneal epithelial cells.

[0069] In the foregoing description, for ease of explanation, numerous details are set forth in order to provide a thorough understanding of the examples. However, it will be apparent to one skilled in the art that these specific details are not required. Therefore, what has been described is merely illustrative of the application of the described examples, and many modifications and variations are possible in light of the above teachings.

[0070] Since the above description provides examples, it should be understood that those skilled in the art may make modifications and changes to the specific examples. Therefore, the scope of the claims should not be limited to the specific examples described herein, but should be interpreted in a manner consistent with the specification as a whole.

Claims

1. Use of a composition for preparing a medicament for alleviating or preventing mitochondrial DNA damage in a subject caused by oxidative stress, the composition comprising: quercetin; ascorbic acid; alpha-lipoic acid; Folate; Vitamin B12; Vitamin E; zeaxanthin; Astaxanthin; Coenzyme Q10; and selenium; in, The composition is formulated for oral administration as two capsules, each capsule containing: 100mg quercetin; 100mg ascorbic acid; 67mg alpha-lipoic acid; 200 μg folate; 25 μg vitamin B12; 67mg vitamin E; 1.25mg zeaxanthin; 2mg astaxanthin; 67mg Coenzyme Q10; and 50μg selenium.

2. The use according to claim 1, wherein The composition is formulated for oral administration.

3. The use according to claim 1, wherein The mitochondrial DNA damage is present in an epithelial cell of the subject; a connective tissue cell of the subject; a muscle tissue cell of the subject; a neural cell of the subject; or any combination thereof.

4. The use according to claim 3, wherein The epithelial cells are cardiovascular epithelial cells or corneal epithelial cells.

5. The use according to claim 3 or 4, wherein Oxidative stress results from exposure to ionizing radiation, or from skin inflammation caused by exposure to UV radiation.

6. The use according to claim 5, wherein The ionizing radiation may come from medical imaging procedures, the release of nuclear material, or air travel.