Skincare formulations containing vitamin E and vitamin B3

Description

BACKGROUND

[0001] Reactive oxygen species (ROS) are free radicals that can react with various cellular proteins in the skin, for example, collagen, elastin and glycosaminoglycans. ROS causes crosslinking of collagen and elastin resulting in wrinkles and decreasing the skin's ability to repair itself. Thus, the skin damage caused by ROS can contribute to the physiology of aging or lead to skin disorders. Antioxidants can scavenge and decrease the excessive ROS produced and mitigate its damage to the cells of different tissues, including skin.

[0002] Furthermore, in the field of skin health, recent consumer trends are requiring products be as simple as possible but still provide acceptable skin health benefits that consumers have grown accustomed to in products with a number of ingredients. However, when using fewer ingredients, there is more of an emphasis on maximizing the results and efficacy of each ingredient in order to delivery anti-oxidant and skin health benefits to a consumer.

[0003] Accordingly, there is a need for a skin health product that is simple but that can also provide, or exceed, the skin health benefits seen in products that contain numerous ingredients that claim to have skin health benefits. WO 2006 / 124992 A1 discloses a skin care composition comprising vitamin E acetate, ergothioneine and niacinamide. The Mintel GNPD Record 1D 8658935 "Energy Boosting Essence Cream", May 2021, XP93066891 discloses an antioxidant cream composition comprising tocopherol, ergothioneine, nicotinamide (niacinamide), arginine and further actives such as hyaluronic acid (as sodium hyaluronate. The Mintel GNPD Record 1D 9177072 "Vital Serum", Nov 2021, XP93066895 discloses a serum composition comprising natural vitamin E, ergothioneine, niacinamide, arginine and further actives such as astaxanthin and hyaluronic acid, as well as adjuvants such as caprylic / capric triglyceride. The Mintel GNPD Record 1D 1859499, "Hydra-Filler Pro-Youth Boosting Moisturiser", Aug 2012, XP2738151 discloses a composition comprising tocopherol, niacinamide, niacin, taurine, arginine, glycine and hyaluronic acid among other active components. WO 01 / 00162 A1) discloses a skin care composition comprising a tocopherol-based antioxidant / radical scavenger and a vitamin B3. The intel GNPD Record ID 1649146, "High Performance Anti-Aging Cream", Oct 2011, XP55636510 discloses a composition comprising tocopherol, ergothioneine and Na-PCA as well as amino acids including arginine and glycine. CN114224779 A discloses an eye repair composition comprising tocopheryl nicotinate and ergothioneine. CN106038361 A discloses a composition comprising 1-4% water-soluble vitamin E, 1-2% sodium pyrrolidone carboxylic acid and 0.5-2% nicotinamide. The Mintel GNPD Record 1D 9285422 "Strength Trainer Peptide Boost Moisturiser", Jan 2022, XP93084654 discloses a cream composition comprising tocopherol, pyrrolidone carboxylic acid, niacinamide, carnosine and further actives such as arginine and glycine. The Mintel GNPD Record 1D 9325230 "Invisible Foundation SPF 60", Jan 2022, XP93084657 discloses a cream composition comprising tocopherol, pyrrolidone carboxylic acid, niacinamide, carnosine and further actives such as arginine and glycine.BRIEF SUMMARY

[0004] The inventors have surprisingly discovered that skin care compositions comprising a combination of vitamin E (ie. alpha-tocopherol) and a thiourea derivative of histidine (ie. ergothioneine) can be combined with vitamin B 3 (ie. niacinamide) in various ratios and amounts, in order to provide unexpectedly enhanced reduction and inhibition of cellular damage caused or related to free radicals (ROS) and environmental damage (by UV rays).

[0005] Niacinamide / Vitamin B3 can function individually to improve barrier function of the skin. However, Niacinamide / Vitamin B3, in combination with alpha-tocopherol and ergothioneine, is believed to unexpectedly enhance the ability to act as an antioxidant by causing a greater decrease in the ROS production than the decrease observed for each of the compounds individually. ROS measurement includes detection and measurement of free radicals in biological systems.

[0006] Further areas of applicability of the present disclosure will become apparent from the detailed description provided hereinafter. In some aspects, the formulations of the disclosure can be used in methods to: revitalize of skin cells, enhance cellular energy in skin cells (e.g., dermal cells), reduce or inhibit cellular damage related to reactive oxygen species and / or UV light, and to detoxify skin cells.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIG. 1 is a bar graph depicting the number of thymine dimer positive cells after application of a non-limiting exemplary compositions and exposure to UV in accordance with an aspect of the invention. FIGS. 2A and 2B are bar graphs depicting the collagen production after application of a non-limiting exemplary compositions and exposure to UV in accordance with an aspect of the invention. FIG. 3 is a bar graph depicting the loricrin production after application of a non-limiting exemplary compositions and exposure to UV in accordance with an aspect of the invention. DETAILED DESCRIPTION

[0008] As used throughout, ranges are used as shorthand for describing each and every value that is within the range. Any value within the range can be selected as the terminus of the range.

[0009] Unless otherwise specified, all percentages and amounts expressed herein and elsewhere in the specification should be understood to refer to percentages by weight relative to the total composition. Unless specified otherwise, the amounts given are based on the active weight of the material.

[0010] The disclosure relates to antioxidant skin care compositions as defined in the claims that effectively treat, prevent and / or reduce free radical damage to the skin of a user. It has been found that Niacinamide / Vitamin B3 in combination with both vitamin E ( alpha-tocopherol) and a thiourea derivative of histidine ( ergothioneine) can provide unexpectedly enhanced protection to the skin from the damage incurred by reactive oxygen species.

[0011] The present invention provides, in one aspect, a skin care composition, comprising from 0.1 to 3 wt.% of vitamin E comprising alpha-tocopherol or topically acceptable salts thereof, from 0.1 to 7 wt.% of an amino acid component, the amino acid component comprising ergothioneine (EGT), wherein the amino acid component further comprises taurine, arginine, and glycine; wherein the skin care composition has a weight ratio of the taurine to the arginine to the glycine (taurine:arginine:glycine) from 85:45:1 to 45:24:1, from 0.7 to 5 wt.% of a vitamin B3 compound comprising niacinamide, and wherein all weight percentages are based on the total weight of the skin care composition as defined in the claims.

[0012] For example, claim 1 refers to a skin care composition comprising: from 0.1 to 3 wt.% of vitamin E comprising alpha-tocopherol or topically acceptable salts thereof, from 0.1 to 7 wt.% of an amino acid component, the amino acid component comprising ergothioneine (EGT), wherein the amino acid component further comprises taurine, arginine, and glycine; wherein the skin care composition has a weight ratio of the taurine to the arginine to the glycine (taurine:arginine:glycine) from 85:45:1 to 45:24:1, from 0.7 to 5 wt.% of a vitamin B3 compound comprising niacinamide, and wherein all weight percentages are based on the total weight of the skin care composition.

[0013] The skin care composition can have a weight ratio of alpha-tocopherol to ergothioneine of 6:1 to 1:6. The skin care composition can have a weight ratio of alpha-tocopherol to niacinamide of from 1:1 to 1:11. The skin care composition can have a weight ratio of ergothioneine to niacinamide of from 1:1 to 1:11. The skin care composition can further comprise carnosine in an amount from 0.1 to 5 wt.%.

[0014] The present disclosure provides, in another aspect, the use of a skin care composition as defined in the claims in a non-therapeutic method of: revitalizing of skin cells, enhancing cellular energy in skin cells, preferably dermal cells, reducing or inhibiting cellular damage related to reactive oxygen species and / or UV light, and detoxifying skin cells as defined in the claims.

[0015] The composition can comprise an oil selected from sunflower seed oil, olive oil, shear butter, jojoba oil, almond oil, grape seed oil, rose hip seed oil, mink oil, castor oil, soybean oil, mineral oil, and a combination thereof, optionally wherein the oil is sunflower seed oil.

[0016] The composition can comprise a thickener. In some embodiments, the thickener comprises a gum, optionally selected from xanthan gum, carrageenan and a combination thereof.

[0017] The composition can comprise a humectant, optionally wherein the humectant is selected from glycerin, sorbitol, and a combination thereof.

[0018] The composition can comprise water.

[0019] The composition can be free or substantially free of sulfate.

[0020] The composition can further comprise a gelling agent, an additional antioxidant, a fragrance, or a combination thereof.

[0021] The composition can be in the form selected from: a serum, a cream, a moisturizer, a mask, a cleanser, a facial make-up, and a balm (e.g., lip balm).

[0022] The composition can be may be any type of personal care composition. In certain embodiments, the composition is any composition that can be formulated into topical skin care formulations suitable for application to skin. Examples of such compositions include, but are not limited to: serums, creams, moisturizers, masks, cleanser (e.g., facial cleansers), a facial make-up, balms (e.g., lip balm), and cosmetics. The composition can comprise a single phase or can be a multi-phase system, for example a system comprising a polar phase and an oil phase, optionally in the form of a stable emulsion. In certain aspects, skin care compositions of the disclosure are formulated in a manner suitable for topically injection.

[0023] The compositions can be liquid, semi-solid or solid. The formulation can be provided in any suitable container, tube, or container with a porous cap, roll-on container, bottle, container with an open end, etc.

[0024] In some aspects, water may be present in the skin care compositions. Water employed in the preparation of commercial personal care compositions should be deionized and free of organic impurities. When it is incorporated into the formulation, water can make up the balance of the compositions and includes 10% to 90%, or 10% to 80%, by weight of the skin care compositions. This amount of water includes the free water which is added plus that amount which is introduced with other materials such as glycerin, sorbitol or any components of the disclosure.

[0025] In some embodiments, the skin care compositions of the disclosure can include one or more pH adjusters. When incorporated in the formulation, the pH adjuster can include, but is not limited to, lactic acid, sodium hydroxide, and / or citric acid.

[0026] In some embodiments, the skin care compositions may further comprise a betaine zwitterionic surfactant. In some aspects, the betaine zwitterionic surfactant may be a C 8 -C 16 aminopropyl betaine, e.g., cocamidopropyl betaine.

[0027] In some embodiments, the skin care compositions described herein may further comprises a non-ionic block copolymer is selected from: Poloxamer 338, Poloxamer 407, Poloxamer, 237, Poloxamer, 217, Poloxamer 124, Poloxamer 184, Poloxamer 185, and a combination of two or more thereof.

[0028] As used herein, the term "effective amount" means the quantity of an active ingredient and / or skin care composition required to provide adequate protection against damage caused or related to reactive oxygen species.

[0029] As used herein, "substantially free" of a material may refer to a composition where the material is present in an amount of less than 0.1 weight %, less than 0.05 weight %, less than 0.01 weight %, less than 0.005 weight %, less than 0.001 weight %, or less than 0.0001 weight % based on a total weight of the composition.

[0030] As used herein, "skin care composition" is meant to refer to a composition for which the intended use can include promotion or improvement of health, cleanliness, odor, appearance, and / or attractiveness of skin. For example, skin care compositions can be in the form of a serum, a cream, a moisturizer, a mask, a cleanser, oil, salve, lotion, gel, ointment, paste, a facial make-up, a toner, an essence, and a balm (e.g., lip balm). Skin care compositions of the disclosure can be applied topically to the skin by a user or consumer. In some aspects, skin care compositions of the disclosure can be applied by injection (e.g., subcutaneous or dermal injection). In certain aspects, skin care composition is intended to refer to a product for use at home or in a professional setting.

[0031] In certain aspects, the skin care compositions of the disclosure are defined in the claims and can additionally comprise one or more further topically active compounds selected from: antibacterial agents, vitamins, medicaments, fragrance materials, antioxidants and other skin-care ingredients.

[0032] As used herein, unless otherwise specified, "Vitamin E" refers to a family of four isomers of tocopherols and four isomers of tocotrienols. All eight isomers of vitamin E have a 6-chromanol ring structure and a side chain. The four tocopherols include fully saturated side chains and include alpha-tocopherol, gamma-tocopherol, beta-tocopherol, and delta-tocopherol. The four tocotrienols include unsaturated side chains and include alpha-tocotrienol, gamma-tocotrienol, beta-tocotrienol, and delta-tocotrienol. As used herein, unless otherwise specified, the term "vitamin E" may refer to any one or more of the eight isomers. For example, as used herein, vitamin E may be or include one or more of alpha-tocopherol, gamma-tocopherol, beta-tocopherol, delta-tocopherol, alpha-tocotrienol, gamma-tocotrienol, beta-tocotrienol, delta-tocotrienol, or any combination thereof. In at least one implementation, the vitamin E includes at least one of the four tocopherols. It should be appreciated that the vitamin E and / or the isomers thereof may be or include natural forms of vitamin E, synthetic forms of vitamin E, or combinations thereof. Any one or more of the isomers of vitamin E may be in the "d" form, the "l" form, or combinations thereof. In some embodiments, vitamin E is vitamin E acetate or Vitamin E succinate. In some embodiments, vitamin E is vitamin E acetate.

[0033] As used herein, "Vitamin C" may be ascorbic acid or derivatives thereof. Ascorbic acid exists as two enantiomers commonly denoted "l" (for "levo") and "d" (for "dextro"). The "l" isomer is the one most often encountered. Ascorbic acid is also referred to as L(+)-ascorbic acid or l-ascorbic acid. The ascorbic acid derivatives may be or include, but are not limited to, L-ascorbic acid, calcium ascorbate, calcium l-ascorbate dihydrate, magnesium ascorbate, potassium ascorbate, magnesium L-ascorbyl phosphate (also referred to as: magnesium ascorbate phosphate or ascorbic acid phosphate magnesium salt), L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate, (+) sodium L-ascorbate, dehydro-l-(+)-ascorbic acid dimer, sodium ascorbyl phosphate (also referred to as: ascorbic acid phosphate sodium salt, sodium l-ascorbyl phosphate, 2-phospho-L-ascorbic acid trisodium salt, L-ascorbic acid 2-phosphate trisodium salt or sodium L-ascorbyl-2-phosphate), ascorbic acid-2-glucoside, ascorbyl dipalmitate, ascorbyl methylsilanol pectinate, ascorbyl stearate, disodium ascorbyl sulfate, ascorbyl 6-palmitate, calcium ascorbyl phosphate, ascorbyl acetate, ascorbyl propionate, ascorbyl stearate, ascorbyl palmitate, ascorbyl dipalmitate, ascorbyl glucoside, ascorbic acid polypeptide, ethyl ascorbyl ether, ascorbyl ethyl silanol pectinate, or the like, or combinations thereof.

[0034] As used herein, "vitamin D" refers to a group of fat-soluble secosteroids responsible for increasing intestinal absorption of calcium, magnesium, and phosphate, and many other biological effects. The two major forms are vitamin D 2 or ergocalciferol, and vitamin D 3 or cholecalciferol. Vitamin D includes vitamin D 1 (mixture of molecular compounds of ergocalciferol with lumisterol), vitamin D 2 (ergocalciferol), vitamin D 3 (cholecalciferol), vitamin D 4 (22-dihydroergocalciferol), and vitamin D 5 (sitocalciferol).

[0035] As used herein, "vitamin K" is a group of compounds with a common chemical structure of 2-methyl-1,4-naphthoquinone.Vitamin K plays a role in blood clotting, bone metabolism, and regulating blood calcium levels. Vitamin K includes vitamin K 1 (phylloquinone) and vitamin K 2 (menaquinone). Vitamin K 2 have unsaturated isoprenyl side chains and are designated as MK-4 through MK-13, based on the length of their side chain.

[0036] Optional ingredients that may be included in the skin care composition of the invention include solvents; water-soluble alcohols such as C 2-8 alcohols including ethanol; glycols including propylene glycol, dipropylene glycol, tripropylene glycol and mixtures thereof; glycerides including mono-, di- and triglycerides; medium to long chain organic acids, alcohols and esters; surfactants including emulsifying and dispersing agents; amino acids including glycine; structurants including thickeners and gelling agents, for example polymers, silicates and silicon dioxide; emollients; fragrances; and colorants including dyes and pigments.

[0037] The skin care compositions of the disclosure may optionally contain emollients in any desired amount to achieve a desired emollient effect. Emollients are known in the art and are used to impart a soothing effect on the skin. Non-volatile emollients are preferable. Classes of non-volatile emollients include non-silicone and silicone emollients. Non-volatile, non-silicone emollients include C 12-15 alkyl benzoate. The non-volatile silicone material can be a polyethersiloxane, polyalkyarylsiloxane or polyethersiloxane copolymer. An illustrative non-volatile silicone material is phenyl trimethicone. Examples include, but are not limited to, PPG-14 butyl ether, PPG-3 myristyl ether, secondary alcohol ethoxylates, stearyl alcohol, stearic acid and salts thereof, glyceryl monoricinoleate, isobutyl palmitate, glyceryl monostearate, isocetyl stearate, sulphated tallow, oleyl alcohol, propylene glycol, isopropyl laurate, mink oil, sorbitan stearate, cetyl alcohol, hydrogenated castor oil, stearyl stearate, hydrogenated soy glycerides, isopropyl isostearate, hexyl laurate, dimethyl brassylate, decyl oleate, diisopropyl adipate, n-dibutyl sebacate, diisopropyl sebacate, 2-ethyl hexyl palmitate, isononyl isononanoate, isodecyl isononanoate, isotridecyl isononanoate, 2-ethyl hexyl palmitate, 2-ethyl hexyl stearate, Di-(2-ethyl hexyl)adipate), Di-(2-ethyl hexyl)succinate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, octacosanol, butyl stearate, glyceryl monostearate, polyethylene glycols, oleic acid, triethylene glycol, lanolin, castor oil, sunflower seed oil, acetylated lanolin alcohols, acetylated lanolin, petrolatum, isopropyl ester of lanolin, fatty acids, mineral oils, butyl myristate, isostearic acid, palmitic acid, PEG-23 oleyl ether, olelyl oleate, isopropyl linoleate, cetyl lactate, lauryl lactate, myristyl lactate, quaternised hydroxy alkyl, aminogluconate, vegetable oils, isodecyl oleate, isostearyl neopentanoate, myristyl myristate, oleyl ethoxy myristate, diglycol stearate, ethylene glycol monostearate, myristyl stearate, isopropyl lanolate, paraffin waxes, glycyrrhizic acid, hydrocyethyl stearate amide. In some embodiments, the composition comprises an oil selected from sunflower seed oil, olive oil, shear butter, jojoba oil, almond oil, grape seed oil, rose hip seed oil, mink oil, castor oil, soybean oil, mineral oil, and a combination thereof. In certain embodiment, the composition comprises sunflower seed oil.

[0038] The skin care compositions of the disclosure may optionally include one or more humectants. Humectants can reduce evaporation and also contribute towards preservation by lowering water activity and can also impart desirable sweetness or flavor to compositions. Illustrative humectants may be or include, but are not limited to, glycerin, propylene glycol, polyethylene glycol, sorbitol, xylitol, or the like, or any mixture or combination thereof. In some embodiment, the humectant is selected from glycerin, sorbitol and a combination thereof. In certain embodiment, the humectant is glycerin.

[0039] The skin care compositions of the disclosure may optionally include thickeners. Illustrative thickeners may be or include, but are not limited to, colloidal silica, fumed silica, a cross-linked polyvinylpyrrolidone (PVP) polymer, cross-linked polyvinylpyrrolidone (PVP), or the like, or mixtures or combinations thereof. In some embodiments, the thickening system includes a cross-linked polyvinylpyrrolidone (PVP) polymer. Illustrative thickeners may also be or include, but are not limited to, carbomers (e.g., carboxyvinyl polymers), carrageenans (e.g., Irish moss, carrageenan, iota-carrageenan, etc.), high molecular weight polyethylene glycols, cellulosic polymers, hydroxyethylcellulose, carboxymethylcellulose, and salts thereof (e.g., CMC sodium), natural gums (e.g., karaya, xanthan, gum arabic, and tragacanth), colloidal magnesium aluminum silicate, or the like, or mixtures or combinations thereof. In some embodiments, the thickener comprises or is a gum, optionally selected from xanthan gum, carrageenan, and a combination thereof.

[0040] The skin care compositions of the disclosure may optionally include one or more gelling agents. Examples of gelling agents include, but are not limited to, waxes, esters of fatty acid and fatty alcohol, triglycerides, partially or fully hydrogenated soybean oil, partially or fully hydrogenated castor oil, other partial or fully hydrogenated plant oils, stearyl alcohol, or other cosmetically acceptable materials, which are solid or semi-solid at room temperature and provide a consistency suitable for application to the skin.

[0041] The skin care compositions of the disclosure may optionally comprise one or more antioxidants. For example, one or more antioxidants may be added to the composition to act as ingredient protectants and for maintenance of long-term stability of the composition. Examples of optional antioxidants include, but are not limited to citric acid, butylated hydroxytoluene, pentaerythrityl tetra-di-t-butyl hydroxyhydrocinnamate.

[0042] The skin care compositions of the disclosure may optionally comprise polymeric materials for thickening, such as polyamides, cellulose derivatives (e.g., hydroxypropylcellulose, hydroxypropyl methyl cellulose, etc.) and natural or synthetic gums, such as polyglycerides including agar, agarose, pectin, or guars or mixtures or combinations thereof. One class of materials worthy of attention for thickening a water-immiscible phase comprises derivatives of hydrolysed starch or other polysaccharides, including in particular esterified dextrins, such as dextrin palmitate. A further class of polymers that is particularly directed to structuring an oil phase containing a silicone oil comprises polysiloxane elastomers. Suspending agents such as silicas or clays such as bentonite, montmorillonite or hectorite, including those available under the trademark Bentone can also be employed to thicken liquid compositions according to the invention. The composition can be thickened with non-polymeric organic gellants, including selected dibenzylidene alditols (e.g., dibenzylidene sorbitol).

[0043] The skin care compositions of the disclosure may optionally include a fragrance. Any fragrance suitable for skin care use may be incorporated into the skin care composition of the disclosure. Fragrances tend to be relatively volatile aroma compounds which are capable of entering the gas phase at skin surface temperature.

[0044] The skin care compositions of the disclosure may be manufactured using methods known in the art. Typically, the ingredients are combined and optionally heated where components need to be melted. The components are mixed. Desirably, volatile materials such as fragrant materials are incorporated in the composition in the latter stages of a mixing cycle in order to avoid volatilization thereof. After mixing, the composition may be poured directly into the dispensers and the container capped to preserve the product until use.EXAMPLES Example 1

[0045] Non-limiting, example skin care compositions (Example Compositions A-D) were prepared according to aspects of the disclosure. The formulations of Example Compositions A-D are provided in Table 1, below (wt.% of active material unless indicated otherwise): Exemplary compositions which do not fall under the scope of the claims are comparative compositions. Table 1IngredientA*B*C**D*Ergothioneine (EGT)0.005%0.006%0.05%0.5%Taurine1.3001.620.651.300Arginine0.6800.8470.340.680Glycine0.020.0250.010.02DL-alpha-tocopherol1.01.250.501.0Vitamin B3 / Niacinamide2.02.51.012.0Sodium PCA1.01.252.011.0Hyaluronic Acid (e.g., 50kDa)1.00.500.25%0.25-1.0Additional Active Ingredients (e.g., carnosine, magnesium sulfate, nicotinamide adenine dinucleotide)1.021.270.26%1.02Multilamellar vesicle (MLV)-49-49Water-q.s. (to balance)-q.s. (to balance)Total 100 100 100 100 * Columns A, B, and D: Amounts given in active weight % unless specified otherwise. ** Column C: Amounts given in raw material weight %. Example 2 - Combinations with alpha-tocopherol, Ergothioneine and Sodium PCA

[0046] In-vitro studies were conducted in a human derived keratinocyte cell culture model. Keratinocytes were treated with exogenous 0.3 wt.% hydrogen peroxide to induce reactive oxygen species (ROS) production. Hydrogen peroxide treated cells were subsequently treated with: alpha-tocopherol, Ergothioneine and Sodium PCA, respectively and in combination. Changes in ROS production were assessed using a ROS Detection Assay Kit and fluorescence microplate reader.Sample preparation: Human derived keratinocytes, plated on 24-well plates

[0047] Raw Materials: alpha-tocopherol Ergothioneine Sodium PCA 30% H 2 O 2 (hydrogen peroxide) Cellular ROS / Superoxide Detection Assay Kit (Abcam - ab139476) Reagents: Medium EpiLife (Life Technologies) Human keratinocyte Growth Supplements (Life Technologies) Stock Solutions: 1 wt.% alpha -tocopherol, was prepared by dissolving 10uL of alpha-tocopherol (100%) in 990 uL of DMSO (100%). 0.1 wt.% alpha -tocopherol, was prepared by dissolving 100uL of alpha-tocopherol (100%) in 900 uL of DMSO (100%). 1 wt.% Ergothioneine (EGT) was prepared by dissolving 10uL of EGT in 990 uL respectively in deionized water. 10 wt.% Sodium PCA (NaPCA) was prepared by dissolving 1mL of Na PCA (50%) in 5mL of deionized water. 1 wt.% Sodium PCA (NaPCA) was prepared by dissolving 1mL of Na PCA (50%) in 9 mL of deionized water. 3 wt.% H 2 O 2 was prepared by dissolving 100uL of H 2 O 2 (30%) in 900 uL of deionized water. 10 wt.% of DMSO, was prepared by dissolving 100uL DMSO (stock 100%) + 900 uL cell culture media. Dilutions:alpha tocopherol: 1 wt.% stock in DMSO, filter-sterilizedDilute20uLof0.1%+980uL=1000uL / 1mLof0.001%Dilute30uLof1%+970uL=1000uL / mLof0.001%Dilute40uLof1%+960uL=1000uL,of0.001%Ergothioneine: 1 wt.% stock in dH2O, filter-sterilizedDilute20uLof1%+980uL=1000uL / 1mLof0.01%Dilute30uLof1%+970uL=1000uL / mLof0.01%Dilute40uLof1%+960uL=1000uL,of0.01%Sodium PCA: 1 wt.% stock in dH2O, filter-sterilizedDilute20uLof1%+980uL=1000uL / 1mLof0.01%Dilute30uLof1%+970uL=1000uL / mLof0.01%Dilute40uLof1%+960uL=1000uL,of0.01%H 2 O 2 : 3 wt.% stock in dH2O, filter-sterilizedDilute200uLof3%+800uL=1000uL / 1mLof0.3%2XDilute300uLof3%+700uL=1000uL / mLof0.01%3XDilute400uLof3%+600uL=1000uL,of0.01%DMSO: 10 wt.% stock in cell culture mediaDilute10uL10%stock+990uLofcellculturemedia=1000uL / 1mLof0.1%Dilute20uL10%stock+980uLofmedia=1000uL / 1mL Table 2 Types of treatment Dilution uLs / well ADMSO1X500uLBα- tocopherol(0.001%)2X250uLCErgothioneine (EGT) (0.01%)2X250uLDSodium PCA (Na PCA) (0.01 %)2X250uLEα-tocopherol (0.001%) + EGT(0.01%) +Na PCA(0.01%)3X167uL eachFDMSO + H 2 O 2 (0.3%)2X250uL + 250uLGα- tocopherol(0.001%) + H 2 O 2 (0.3%)2X250uL + 250uLHEGT (0.01 %) + H 2 O 2 (0.3%)2X250uL + 250uLINa PCA (0.01 %) + H 2 O 2 (0.3%)2X250uL + 250uLJα- tocopherol (0.001%) + EGT(0.01%) +Na PCA(0.01%) + H 2 O 2 (0.3%)4X125uL each Cell culture treatment 500 uL of each treatment sample was directly applied to the 24-well plates and incubated for 30 mins. Post incubation: each well was washed with the wash buffer (1X) provided. 250uL of green Fluorescent (GF) dye containing media (2.5uL of GF dye / 250 uL of media) was then added to each well and incubate for 60 mins. Post incubation, Fluorescence was read on a microplate reader at 490 / 525. Test Products:

[0048] Test samples are listed in Table 3. Table 3Test ProductH 2 O 2 (0.3%)DescriptionADMSO+0.1% of DMSOFDMSO+0.1% of DMSO + 0.3% H 2 O 2 in untreated mediaGAlpha-tocopherol (0.001%)+Alpha-tocopherol (0.001%) + 0.3% H 2 O 2 in untreated mediaHEGT (0.01%)+EGT (0.01%) + 0.3% H 2 O 2 in untreated mediaINa PCA (0.01%)+Na PCA (0.01%) + 0.3% H 2 O 2 in untreated mediaJAlpha-tocopherol (0.001%) + EGT (0.01%) + Na PCA (0.01%)+Alpha-tocopherol (0.001%) + EGT (0.01%) + Na PCA (0.01%) + 0.3% H 2 O 2 in untreated media

[0049] Results: 0.3 wt.% H 2 O 2 treatment results in a significant increase in ROS produced by the human derived keratinocytes. 0.001 wt.% of α- tocopherol + 0.01 wt.% EGT + 0.01% Sodium PCA collective treatment in human derived keratinocyte culture resulted in a greater than 82% decrease in ROS generated (relative to the 0.3 wt.% of H 2 O 2 treatment). This decrease in ROS with a combinatorial treatment of α- tocopherol, EGT and sodium PCA is unexpected and significantly greater than the decrease observed by individual treatment by 0.001 wt.% of any of: α- tocopherol (70.72% decrease), 0.01 wt.% EGT (74.5% decrease) and 0.01 wt.% Sodium PCA (37.9%), individually.

[0050] Experiments were performed in three replicas. The results are shown in Tables 4 and 5 below. Table 4 - Significant Decrease in ROS generated in keratinocyte by antioxidant treatment (Fold Change (relative to untreated))TreatmentP ValuesFold ChangeA vs FDMSO vs. 0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media*** p < 0.00015.95-fold increaseF vs G0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media vs. Alpha-tocopherol (0.001%)*** p < 0.00014.21-fold decreaseFvsH0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media vs. EGT (0.01%)*** p < 0.00014.43-fold decreaseF vs I0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media vs. Na PCA (0.01%)*** p < 0.00012.25-fold decreaseF vs J0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media vs. Alpha-tocopherol (0.001%) + EGT (0.01%) + Na PCA (0.01%)*** p < 0.00014.92-fold decrease Table 5 - Decrease in ROS generated in keratinocyte by antioxidant treatment (relative to untreated TreatmentH 2 O 2 (0.3%)% DecreaseADMSO-87.77%FDMSO vs. 0.1 wt.% of DMSO + 0.3 wt.% H 2 O 2 in untreated media+0%GAlpha-tocopherol (0.001%)+70.72%HEGT (0.01%)+74.5%INa PCA (0.01%)+37.88%JAlpha-tocopherol (0.001%) + EGT (0.01%) + Na PCA (0.01%)+82.78%

[0051] The results demonstrate that a combination of α- tocopherol, Ergothioneine (EGT) and Sodium PCA exhibit a significant decrease in ROS production in human derived keratinocyte cell culture. This combination shows an unexpected impact in decreasing ROS production in keratinocytes (treated with 0.3% hydrogen peroxide) relative to each treatment alone.Example 3 - Combinations with alpha-tocopherol, Ergothioneine and Niacinamide

[0052] In-vitro studies were conducted in a human derived keratinocyte cell culture model. Keratinocytes were treated with exogenous 0.3 wt.% hydrogen peroxide to induce ROS production. Hydrogen peroxide treated cells were then treated with: alpha-tocopherol, ergothioneine and niacinamide, respectively and in combination. Changes in ROS production were then assessed using a ROS Detection Assay Kit and fluorescence microplate reader.Sample preparation: Human derived keratinocytes, plated on 24-well plates

[0053] Raw Materials: alpha-tocopherol Ergothioneine Niacinamide 30% H 2 O 2 (hydrogen peroxide) Cellular ROS / Superoxide Detection Assay Kit (Abcam - ab139476) Reagents: Medium EpiLife (Life Technologies) Human keratinocyte Growth Supplements (Life Technologies) Stock Solutions: 1 wt.% alpha -tocopherol, is prepared by dissolving 10uL of alpha-tocopherol (100%) in 990uL of DMSO (100%). 1 wt.% Ergothioneine (EGT) and Niacinamide are prepared by dissolving 10mg / ml, followed by filter sterilization. 3 wt.% H 2 O 2 is prepared by dissolving 100uL of H 2 O 2 (30%) in 900uL of deionized water. 10% of DMSO, is prepared by dissolving 100uL DMSO (stock 100%) + 900uL cell culture media. Dilutions:alpha tocopherol: 1 wt.% stock in DMSO, filter-sterilizedDilute10uLof1%+990uL=1000uL / 1mLof0.01%Dilute20uLof1%+980uL=1000uL / 1mLof0.01%Dilute30uLof1%+970uL=1000uL / mLof0.01%Dilute40uLof1%+960uL=1000uL,of0.01%Ergothioneine: 1 wt.% stock in distilled water (dH2O), filter-sterilizedDilute10uLof1%+990uL=1000uL / 1mLof0.01%Dilute20uLof1%+980uL=1000uL / 1mLof0.01%Dilute30uLof1%+970uL=1000uL / mLof0.01%Dilute40uLof1%+960uL=1000uL,of0.01%Niacinamide: 1 wt.% stock in dH2O, filter-sterilizedDilute10uLof1%+990uL=1000uL / 1mLof0.01%Dilute20uLof1%+980uL=1000uL / 1mLof0.01%Dilute30uLof1%+970uL=1000uL / mLof0.01%Dilute40uLof1%+960uL=1000uL,of0.01%H 2 O 2 : 3 wt.% stock in dH2O, filter-sterilizedDilute10uLof1%+990uL=1000uL / 1mLof0.03%Dilute200uLof3%+800uL=1000uL / 1mLof0.3%Dilute300uLof3%+700uL=1000uL / mLof0.01%Dilute400uLof3%+600uL=1000uL,of0.01%DMSO: 10 wt.% stock in cell culture mediaDilute10uL10%stock+990uLofcellculturemedia=1000uL / 1mLof0.1%Dilute20uL10%stock+980uLofmedia=1000uL / 1mL Types of treatment Dilution uLs / well ADMSO1X500uLBα- tocopherol(0.01%)1X500uLCErgothioneine (EGT) (0.01 %)1X500uLDNiacinamide (0.01%)1X500uLEα- tocopherol (0.01%) + EGT(0.01%) +Nicotinamide(0.01%)3X167uL + 167uLFDMSO + H 2 O 2 (0.3%)2X250uL + 250uLGα- tocopherol(0.01%) + H 2 O 2 (0.3%)2X250uL + 250uLHEGT (0.01%) + H 2 O 2 (0.3%)2X250uL + 250uLINiacinamide (0.01%) + H 2 O 2 (0.3%)2X250uL + 250uLJα- tocopherol (0.01%) + EGT(0.01%) +Nicotinamide(0.01%) + H 2 O 2 (0.3%)4X125uL + 125uL Cell culture treatment 500 uL of each treatment sample was directly applied to the 24-well plates and incubated for 30 mins. Post incubation: each well was washed with the wash buffer (1X) provided. 250uL of green Fluorescent (GF) dye containing media (2.5uL of GF dye / 250uL of media) was then added to each well and incubate for 60 mins. Post incubation, Fluorescence was read on a microplate reader at 490 / 525. Test Products:

[0054] Test samples are listed in Table 6. Table 6Test Product (wt.%)H 2 O 2 (0.3 wt.%)Description (wt.%)ADMSO-0.1% + DMSOFDMSO+0.1% + DMSO + 0.3% of H 2 O 2 (0.3%) in untreated mediaGAlpha-tocopherol (0.01%)+Alpha-tocopherol (0.01%) + 0.3% of H 2 O 2 (0.3%) in untreated mediaHEGT (0.01%)+EGT (0.01%) + 0.3% of H 2 O 2 (0.3%) in untreated mediaINiacinamide (0.01%)+Niacinamide (0.01%) + 0.3% of H 2 O 2 (0.3%) in untreated mediaJAlpha-tocopherol (0.01%) + EGT (0.01%) + Niacinamide (0.01%)+Alpha-tocopherol (0.01%) + EGT (0.01%) + Niacinamide + 0.3% of H 2 O 2 (0.3%) in untreated media

[0055] Results: 0.3 wt.% H 2 O 2 treatment results in a significant increase in ROS produced by the human derived keratinocytes. 0.01 wt.% of α- tocopherol + 0.01% EGT + 0.01% Niacinamide collective treatment on human derived keratinocyte culture results in greater than 36% decrease in ROS generated relative to the 0.3 wt.% of H 2 O 2 treatment. This decrease in ROS by combinatorial treatment of the three compounds is significantly greater than the decrease observed by individual treatment by 0.01 wt.% of α-tocopherol, 0.01 wt.% EGT and 0.01 wt.% Niacinamide, respectively.

[0056] Experiments were performed in three replicas. The results are shown in Tables 7 and 8 below. Table 7 - Significant Decrease in ROS generated in keratinocyte by antioxidant treatment (Fold Change (relative to untreated))TreatmentP ValuesFold ChangeA vs FDMSO vs. 0.1% of DMSO + 0.3% H 2 O 2 in untreated media** p < 0.00251.5-fold increaseFvsG0.1% of DMSO + 0.3% H 2 O 2 in untreated media vs. Alpha-tocopherol (0.001%)*p < 0.01390.4-fold decreaseFvsH0.1% of DMSO + 0.3% H 2 O 2 in untreated media vs. EGT (0.01%)Not significant0.31-fold decreaseF vs I0.1% of DMSO + 0.3% H 2 O 2 in untreated media vs. Niacinamide (0.01%)Not significant0.24-fold decreaseF vs J0.1% of DMSO + 0.3% H 2 O 2 in untreated media vs. Alpha-tocopherol (0.001%) + EGT (0.01%) + Niacinamide (0.01%)**p < 0.00380.54-fold decrease Table 8 - Decrease in ROS generated in keratinocyte by antioxidant treatment TreatmentH 2 O 2 (0.3 wt.%)% DecreaseADMSO-33.33%FDMSO vs. 0.1% of DMSO + 0.3% H 2 O 2 in untreated media+0%GAlpha-tocopherol (0.001%)+20.79%HEGT (0.01%)+15.91%INa PCA (0.01%)+25.75%JAlpha-tocopherol (0.001%) + EGT (0.01%) + Niacinamide (0.01%)+36.13%

[0057] The results demonstrate that a combination of α-tocopherol, ergothioneine (EGT), and niacinamide provided a significant decrease in ROS production in human derived keratinocyte cell culture. This combination also shows an unexpected impact in decreasing ROS production in keratinocytes (treated with 0.3 wt.% hydrogen peroxide) relative to each treatment alone.Example 4 - Comparison of various ratios of Alpha-tocopherol, EGT, Niacinamide

[0058] In further experiments various weight ratios of alpha-tocopherol to EGT to niacinamide (alpha-tocopherol: EGT: niacinamide) were tested to determine efficacy in reducing ROS production in human derived keratinocyte cell culture. The weight ratio of 1:1:3 for alpha-tocopherol to EGT to niacinamide (alpha-tocopherol: EGT: niacinamide) showed an unexpected enhancement in decreasing ROS production in keratinocytes (treated with 0.3 wt.% hydrogen peroxide) relative to other ratios of the same ingredient combination, including 1:1:1 (alpha-tocopherol: EGT: niacinamide).

[0059] Solutions with various ratios of Alpha-tocopherol: EGT: Niacinamide were tested in in-vitro studies conducted in a human derived keratinocyte cell culture model. Keratinocytes were treated with exogenous 0.3 wt.% hydrogen peroxide to induce ROS production.

[0060] Stock Solutions: 1 wt.% α-tocopherol, was prepared by dissolving 10uL of a-tocopherol (100%) in 990uL of DMSO (100%). 1 wt.% niacinamide was prepared by dissolving in deionized water at 10mg / mL, followed by filter sterilization. 1 wt.% ergothioneine was prepared by dissolving 10uL of BioYouth EGT in 990 uL of deionized water followed by filter sterilization 3 wt.% H 2 O 2 was prepared by dissolving 100uL of H2O2(30%) in 900uL of deionized water. 10 wt.% of DMSO was prepared by dissolving 100uL DMSO (stock 100%) + 900uL cell culture media.

[0061] Dilutions α-tocopherol: 1 wt.% stock in DMSO, filter-sterilized. Dilute 10 uL of 1 % + 990 uL = 1000 uL / mL of 0.01 % . Niacinamide: 1 wt.% stock in distilled water (dH2O), filter-sterilized. Dilute 10 uL of 1 % + 990 uL = 1000 uL / mL of 0.01 % . Ergothioneine: 1 wt.% stock in dH2O, filter-sterilized. Dilute 10 uL of 1 % + 990 uL = 1000 uL / mL of 0.01 % . H2O2: 3 wt.% stock in dH2O, filter-sterilized. Dilute 100 uL of 3 % + 900 uL = 1000 uL / 1 mL of 0.3 % .

[0062] The evaluated plate layout, which had different ratios of components is shown in Table 9, below. Table 9Untreated (400uL of media)Untreated (400uL of media)Untreated (400uL of media)0.3 wt.% H 2 O 2 (400uL of 0.3% H2O2)0.3 wt.% H 2 O 2 (400uL of 0.3% H2O2)0.3 wt.% H 2 O 2 (400uL of 0.3% H2O2)0.01 wt.% α tocopherol + EGT + Niacinamide 1:1:1 (100uL each = 300ul) +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT + Niacinamide 1:1:1 (100uL each = 300ul) +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT + Niacinamide 1:1:1 (100uL each = 300ul) +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT + Niacinamide 1:2:1 (75uL:150uL:75 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT + Niacinamide 1:2:1 (75uL:150uL:75 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT + Niacinamide 1:2:1 (75uL:150uL:75u L)=300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 2:1:1 (150uL:75uL:75 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 2:1:1 (150uL:75uL:75 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 2:1:1 (150uL:75uL:75 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:1:2 (75uL:75uL:150 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:1:2 (75uL:75uL:150 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:1:2 (75uL:75uL:150u L) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 3:1:1 (180uL:60uL:60 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 3:1:1 (180uL:60uL:60 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:3:1 (60uL:180uL:60 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:3:1 (60uL:180uL:60 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:1:3 (60uL:60uL:180 uL) = 300uL +0.3% H 2 O 2 (100ul)0.01 wt.% α tocopherol + EGT +Niacinamide 1:1:3 (60uL:60uL:180u L)= 300uL +0.3% H 2 O 2 (100ul) Table 10 - Decrease in ROS generated in keratinocyte by antioxidant treatment TreatmentFold Change (relative to untreated)Untreated10.3% H 2 O 2 in untreated media2.3-fold increase1:1:1 (Alpha-tocopherol: EGT: Niacinamide)1.4-fold decrease2:1:1 ((Alpha-tocopherol: EGT: Niacinamide)1.4-fold decrease1:2:1 ((Alpha-tocopherol: EGT: Niacinamide)1.45-fold decrease1:1:2 ((Alpha-tocopherol: EGT: Niacinamide)1.47-fold decrease1:3:1 ((Alpha-tocopherol: EGT: Niacinamide)1.46-fold decrease3:1:1 ((Alpha-tocopherol: EGT: Niacinamide)1.39-fold decrease1:1:3 ((Alpha-tocopherol: EGT: Niacinamide)1.57-fold decrease Example 5 - Combinations with alpha-tocopherol, Ergothioneine and Niacinamide and UV protection

[0063] Exposure to UV radiation can cause an increase in photolesions in the form of thymine dimers. For example, due to exposure to UV radiations adjacent thymine residues can become linked covalently resulting in dimers. These dimers can be replicated as a single base causing mutations. Additionally, exposure to UV radiations can possibly decrease enzymes such as catalase. Catalase is believed to convert reactive oxygen species such as hydrogen peroxide to water and oxygen, reducing its toxic effects. Additionally, exposure to UV is believed to possibly decrease skin barrier protein (such as loricrin) and extracellular matrix proteins (such as collagen).

[0064] The outermost layer, epidermis of the skin, serves the function of forming a protective barrier against external environmental stresses, chemical damage and bacterial infection. The skin barrier comprises a number of proteins including Loricrin. Loricrin is a major protein present in the terminally differentiated cornified envelope and contributes to the protective barrier function of the stratum corneum.

[0065] The extracellular matrix (ECM) is composed of a number of different macromolecules including collagen, elastin, fibronectin and laminin, all believed to be organized in a tissue-specific manner. These protein components link together to form structurally stable tissue supporting structures.

[0066] In these Examples, Genoskin human skin explants were either 1) left untreated; 2) exposed to UV (A+B) at 20J / cm2); 3) treated with placebo followed by UV exposure; or 4) treated with a mixture of alpha-tocopherol, ergothioneine (EGT) and niacinamide, in a weight ratio of alpha-tocopherol to EGT to niacinamide of 1:1:3 ratio, followed by UV exposure. Changes in the number of thymine dimer-positive cells / field of view, the amount of catalase expression, the fold changes in loricrin and extracellular matrix protein collagen expression were measured for each treatment.

[0067] From the data, the combined mixture of alpha-tocopherol, ergothioneine (EGT), and niacinamide in a weight ratio of alpha-tocopherol to EGT to niacinamide of 1:1:3 is believed to benefit the skin by: decreasing mutation causing changes in skin cells, restoring protective enzymes in the skin, and helping barrier and extracellular matrix formation to mimic that of untreated / nascent skin.Method: Tissue treatment:

[0068] In vitro studies were conducted using Genoskin human skin explants. The skin explants were first normalized at least one hour before treatments with 1 mL media and incubated at a temperature of 37°C with 5% CO 2 before being left untreated, treated with placebo, or treated with a mixture of alpha-tocopherol, EGT, and niacinamide at a weight ratio of alpha-tocopherol to EGT to niacinamide of 1:1:3. The skin explants being treated with the placebo or mixture of alpha-tocopherol, EGT, and niacinamide received an application of the placebo or the mixture alpha-tocopherol, EGT, and niacinamide once a day for 5 days. On the sixth day, the skin explants receiving the treatment were then exposed to UVA+UVB at 20 J / cm2. Following the UV exposure, the tissue samples were then treated with either the placebo or the mixture and incubated for 24 hours at a temperature of 37°C with 5% CO 2 . The tissue samples were then collected for either RNA extraction and analysis or fixed with 4 wt.% PFA, followed by paraffin embedding processes.Immunofluorescent staining:

[0069] The PFA-fixed paraffin-embedded tissues were de-waxed by heat and xylene treatment followed by hydration of the tissue by a gradient of ethanol ranging from 100% to 50% followed by water. The tissue slides then go through the antigen retrieval step using citric acid-based solution (pH 6.0) for 30 minutes, during which tissue slides were immersed in the solution and heated using a commercially available hot plate stirrer. The tissue slides were then cooled down to a temperature of 60°C, and washed 1X PBS. Following this, the tissue samples were blocked for 1 hour 30 minutes at room temperature in PBS containing 5 wt.% goat serum and 0.1 wt.% triton x100.

[0070] Primary antibody (anti-rabbit Catalase) incubations occurred overnight at a temperature of 4°C. Post primary antibody treatment, the tissue slides were washed in PBS and incubated with fluorescently labelled secondary antibody (Alexa Fluor 488 Dye, ThermoFisher Scientific, A11006, 1:1000) for 1 hour 30 minutes at room temperature. Following the secondary antibody treatment, the slides were washed with PBS followed by air-drying and mounting them with Prolong Gold antifade medium containing DAPI nuclear counterstain (vectashield antifade mounting media with DAPI) (vector laboratories, H1200). Images were collected by a microscope, EVOS FL Auto (Life technologies).Immunohistochemistry and thymine dimer

[0071] The tissue sections were heated for 1 hour at a temperature of 60°C followed by hydration with Xylene, 100% - 30% Ethanol and deionized water. The tissue slides then go through the antigen retrieval step using citric acid-based solution (pH 6.0) for 30 minutes, during which the tissue slides were immersed in the solution and heated using a commercially available hot plate stirrer. The tissue slides were then cooled down to 60°C, and washed 1X PBS. Following this, the tissue samples were blocked for 1 hour 30 minutes at room temperature in PBS containing 5 wt.% goat serum and 0.1 wt.% triton x100. For DNA denaturation: the tissue sections were placed in 2N HCl for 30 minutes at a temperature 37°C followed by wash and hydrogen peroxide block: 3 wt.% hydrogen peroxide (1 mL 30% hydrogen peroxide + 9 mL PBS) for 15 min at RT. Primary Antibody was then applied and incubated for 1 hour at room temperature (Thymine Dimer (mouse monoclonal I1192- 1.5ul in 1.5mL) 1:1000). This was followed by a secondary antibody treatment of Biotinylated Universal Antibody incubated for 30 min at room temperature. RTU Vectastain Elite ABC Reagent was then applied and incubated for 30 min followed by ImmPACT NovaRED (5mL Diluent, 3 drops #1, 2drops #2, 2drops #3, 2 drops #4) and incubate for 2-15 min, at room temperature respectively. Finally, the tissues undergo counterstain with hematoxylin and mounting.RT-QPCR:

[0072] RNA was extracted from RNAlater stabilized tissues using a homogenizer (Kinematica, Bohemia, NY) and RNeasy Plus Mini Kit (Qiagen) according to the manufacturer's protocol. Extracted RNAs was quantitated using NanoDrop One (ThermoFisher Scientific). Based on the RNA concentrations, cDNA synthesis was performed using Maxima First Strand cDNA Synthesis kit for RT-qPCR with dsDNase (ThermoFisher) and gene expression analysis was performed using Taqman ™< array cards using QuantStudio 7 Flex (ThermoFisher). Fourty-eight skin specific targets and endogenous controls were selected for analysis. Relative quantitations from treatments were calculated and compared to UV only samples or untreated. A t-test was used to evaluate statistical significance and p-value was corrected by Benjanmini-Hochberg false discovery rate using the Thermo Fisher Cloud app.

[0073] Reagents: RNeasy Mini Kit (QIAGEN) Thermo Scientific Maxima H Minus First Strand cDNA Synthesis Kit TaqMan ™< Fast Advanced Master Mix

[0074] Results:

[0075] The tissues were either untreated, treated with UV alone, treated with placebo following UV, or treated with the antioxidant mix of alpha-tocopherol, EGT, and niacinamide in a weight ratio of alpha-tocopherol to EGT to niacinamide of 1: 1:3 followed by UV. Post the one week treatment, the tissues were fixed and sectioned and immunohistochemically labeled with an antibody against a thymine dimer.

[0076] The results demonstrated that with UV light treatment there was a significant increase in thymine dimer positive cells in tissues. However, the antioxidant mixture treatment exhibited a significant reduction in the effect of UV on the tissues. Data is presented in Table 11 and FIG. 1. Table 11TreatmentAverage number of thymine dimer positive cells relative to the field of viewUntreated0Untreated + UV Light43.6 *Placebo22.4Alpha-tocopherol + EGT + Niacinamide (1:1:3)***3.8 *** p < 0.001 vs. "untreated" ** p < 0.001 vs. "untreated + UV Light" *** Based on raw material weights.

[0077] The tissues were either untreated, treated with UV alone, treated with placebo followed by UV light or treated with the antioxidant mix followed by UV light. After a one week treatment, the tissues were collected and their RNA extracted followed by cDNA synthesis. Quantitative Real Time PCR (qRT-PCR) was performed to evaluate changes in the genetic expression of Extracellular matrix protein, such as Collagen (Col1A and Col3A), and skin barrier proteins (loricrin). A summary of the results are presented in Tables 12, 13, and 14 and FIGS. 2A, 2B, and 3. Table 12: Collagen 1A expressionTreatmentFold Change Relative to UntreatedUntreated1Untreated + UV Light0.688Placebo1.985Alpha-tocopherol + EGT + Niacinamide (1:1:3)*2.635 Table 13: Collagen 3A expression TreatmentFold Change Relative to UntreatedUntreated1Untreated + UV Light0.729Placebo1.428Alpha-tocopherol + EGT + Niacinamide (1:1:3)*2.52 Table 14: Loricrin expression TreatmentFold Change Relative to UntreatedUntreated1Untreated + UV Light0.337Placebo0.412Alpha-tocopherol + EGT + Niacinamide (1:1:3)*0.539

[0078] The tissues were either untreated, treated with UV alone, treated with placebo followed by UV light or treated with the antioxidant mix followed by UV light. Post the one-week treatment, the tissues were fixed and sectioned and immunohistochemically labeled with an antibody against catalase. For the UV treatment tissues, there was a significant decrease in catalase expression in tissues. However, the antioxidant mixture (alpha-tocopherol, EGT, and niacinamide) treatment provided a significant decrease in the effect of UV light on the tissues. A summary of the results from the immunohistochemistry labelling are described in Table 15. Table 15: Immunofluorescent Staining and Catalase ExpressionTreatmentMean grey intensity of catalase expressionUntreated14.7Untreated + UV Light11.7Placebo10.6Alpha-tocopherol + EGT + Niacinamide (1:1:3)*18.7 Example 5

[0079] Non-limiting, example skin care compositions (Example Compositions E-H) were prepared according to aspects of the disclosure. The formulation for Example Compositions EH are shown in Table 16. Table 16US INCI Compound NameEx. E (wt.%)Ex. F (wt.%)Ex. G (wt.%)Ex. H (wt.%)Niacinamide0.408212Taurine0.2651.30.651.3Carnosine0.20410.51PCA0.20410.51Arginine0.1390.680.340.68Hydrolyzed Hyaluronic acid0.05110.51Glycine0.0040.020.010.02Magnesium Sulfate0.0020.010.0050.01Nicotinamide Adenine Dinucleiotide0.0020.010.0050.01EGT0.0010.0050.00250.005Tocopherol0.20.50.51Encapsulation (made by Effervescence)0.80.80.80.8Water8.2110.51Mannitol0.2030.9950.49750.995Pentylene glycol0.15Citric acid0.13Sodium Benzoate0.016Benzoic acid0.01BASE89.00189.6894.1989.18

Claims

1. A skin care composition comprising: from 0.1 to 3 wt.% of vitamin E comprising alpha-tocopherol or topically acceptable salts thereof, from 0.1 to 7 wt.% of an amino acid component, the amino acid component comprising ergothioneine (EGT), wherein the amino acid component further comprises taurine, arginine, and glycine; wherein the skin care composition has a weight ratio of the taurine to the arginine to the glycine (taurine:arginine:glycine) from 85:45:1 to 45:24:1, from 0.7 to 5 wt.% of a vitamin B3 compound comprising niacinamide, and wherein all weight percentages are based on the total weight of the skin care composition.

2. The skin care composition according to claim 1, wherein the skin care composition has a weight ratio of alpha-tocopherol to ergothioneine of 6:1 to 1:6.

3. The skin care composition according to claim 1 or claim 2, wherein the skin care composition has a weight ratio of alpha-tocopherol to niacinamide of from 1:1 to 1:11.

4. The skin care composition according to any one of claims 1 to 3, and / or wherein the skin care composition has a weight ratio of ergothioneine to niacinamide of from 1:1 to 1:11.

5. The skin care composition according to any one of claims 1 to 4, wherein the skin care composition further comprises carnosine in an amount from 0.1 to 5 wt.%.

6. Use of a skin care composition of any preceding claim in a non-therapeutic method of: revitalizing of skin cells, enhancing cellular energy in skin cells, preferably dermal cells, reducing or inhibiting cellular damage related to reactive oxygen species, and detoxifying skin cells.