Topical compositions and methods of use thereof

Abstract

The present invention relates generally to methods and compositions effective for alleviating skin inflammation in a subject. In some embodiments, the methods and compositions disclosed herein include topical application of a cosmetic formulation, where the cosmetic formulation comprises at least one transient receptor potential channel vanillic acid subtype 1 (TRPV-1) antagonist. In some embodiments, the compositions and / or methods include a plankton extract (e.g., a saccharide isomer), a reed (Phragmites communis, Common Reed) extract, a Poria cocos (Poria cocos, Smilax glabra root) extract, and / or a Cucurbita pepo, pumpkin seed extract, and in some embodiments, the compositions and / or methods include a plankton extract (e.g., a saccharide isomer), a Phragmites communis (Phragmites communis, Common Reed) extract, a Poria cocos (Poria cocos, Smilax glabra root) extract, and / or a Cucurbita pepo, pumpkin seed extract.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application Serial No. 63 / 336,622, filed on April 29, 2022, which is incorporated herein by reference in its entirety. background

[0002] A. Field of the Invention

[0003] The present invention relates generally to the field of topical skin compositions. More specifically, the present invention relates to the use of topical skin compositions for reducing skin irritation caused by activation of transient receptor potential channel vanilloid subtype 1 (TRPV-1) agonists (eg, retinol activation).

[0004] B. Related technical description

[0005] Aging, chronic exposure to adverse environmental factors, malnutrition, fatigue, etc. can alter the visual appearance, physical properties, or physiological functions of the skin in visually undesirable ways. The most significant and obvious changes include the appearance of fine lines and wrinkles, loss of elasticity, increased sagging, decreased firmness, loss of color uniformity or tone, rough surface texture, and mottled pigmentation. As the skin ages or is subjected to chronic environmental insults, the skin undergoes subtle but measurable changes, including a general decrease in cell and tissue viability, reduced cell replication rates, decreased skin blood flow, decreased water content, accumulation of structural and functional errors, alterations in the normal regulation of common biochemical pathways, and a decrease in the skin's ability to remodel and repair itself. Many changes in the skin's appearance and function are caused by changes in the outer epidermal layer of the skin, while others are caused by changes in the dermis.

[0006] Previous attempts to improve the visual appearance of the skin using known skin active ingredients have been shown to have various disadvantages, such as skin irritation and prolonged recovery period. Retinol (vitamin A) is one of the first FDA-approved anti-aging therapeutics, which is used in cosmetic preparations for treating skin photodamage and delaying aging. Retinol binds to different receptors to regulate various cellular processes, such as, but not limited to, keratinocyte differentiation, blocking the transport of melanin to epidermal cells, inhibiting matrix metalloproteinases (MMPs), and / or stimulating the synthesis of collagen fibers in fibroblasts. (M Zasada et al.; 2019). In addition, retinol can also increase the excitability of nociceptors (sensory neurons), and this increase in excitability can lead to increased tissue sensitivity and / or irritation. Irritation caused by retinoids is the main reason why subjects do not comply with the treatment regimen and / or stop using retinol before improving the appearance of the skin. In addition, excessive use of retinol can also exacerbate skin dryness and / or burning sensations. Published literature describes that up to 12% of subjects reported adverse events (AEs) when using retinol. Skin inflammation may be a direct response to noxious chemosensory stimuli. Certain cells, such as epidermal keratinocytes, melanocytes, and / or fibroblasts, may also release cytokines in response to noxious stimuli, which, in addition to other inflammatory effects, can sensitize peripheral nociceptive fibers and produce neurogenic inflammation. Previous studies have shown that retinol activates transient receptor potential channel vanilloid subtype 1 (TRPV-1), which is a stimulatory receptor for capsaicin (S Yin et al.; 2013).

[0007] It is shown herein that a combination of one or more TRPV-1 antagonists (e.g., saccharide isomers, pumpkin seed extract, common reed extract, and / or China root extract) can be used to inhibit TRPV-1 mediated calcium flux and related tissue irritation, such as irritation caused by anti-aging compounds (e.g., irritation caused by retinoids). In addition, the inventors show that such TRPV-1 antagonists can be optionally combined with one or more pro-inflammatory cytokine inhibitors, and / or optionally combined with one or more anti-aging compound tolerance improvement regimens to further increase an individual's tolerance to a treatment regimen that includes irritating anti-aging compounds (e.g., hydroquinone, resorcinol, trichloroacetic acid, carboxylic acids, dicarboxylic acids, beta-hydroxy acids, and / or alpha-hydroxy acids). Summary of the invention

[0008] The methods and compositions disclosed herein overcome the deficiencies in the art by recognizing the importance of treatment regimen compliance and providing improved treatment regimen compliance methods and compositions. More specifically, the inventors have identified a group of compounds that are TRPV-1 antagonists and inhibitors of related cellular calcium fluxes. In certain embodiments, the disclosed TRPV-1 antagonists can be used alone, together, or in combination with other compounds to produce compositions and methods of use suitable for use in the fields of beauty, nutrition, and / or medicine.

[0009] In certain embodiments, the compositions disclosed herein are suitable for combating various signs of aging, while reducing adverse events associated with activation of TRPV-1 by anti-aging compounds (e.g., TRPV-1 agonist anti-aging compounds), and / or while reducing adverse events associated with cytokine release induced by anti-aging compounds. In certain embodiments, the anti-aging compound may be, but is not limited to, hydroquinone, resorcinol, trichloroacetic acid, carboxylic acid, dicarboxylic acid, β-hydroxy acid, and / or α-hydroxy acid. In certain embodiments, α-hydroxy acid is citric acid, glycolic acid, lactic acid, malic acid, mandelic acid, and / or tartaric acid. In certain embodiments, β-hydroxy acid is salicylic acid, salicylic acid derivatives or metabolites, β-hydroxybutyric acid, tropic acid, and / or tresocaic acid. In certain embodiments, the agonist of TRPV-1 is retinol, a retinol precursor, a retinol derivative, and / or a retinol metabolite. In certain embodiments, the agonist of TRPV-1 is resorcinol. In certain embodiments, the agonist of TRPV-1 is salicylic acid.

[0010] In certain embodiments, the compositions and / or methods disclosed herein are useful for reducing deep lines and wrinkles, evening out uneven skin tone, brightening skin tone, increasing skin radiance, reducing photodamage, increasing skin elasticity, increasing skin firmness, reducing skin sagging, reducing facial volume loss, increasing skin barrier function, inhibiting skin antioxidant capacity, increasing skin collagen expression, increasing skin elastin expression, increasing skin laminin expression, inhibiting MMP-1, inhibiting MMP-3, inhibiting MMP-9, inhibiting proinflammatory cytokines, inhibiting skin elastase expression, increasing skin fibronectin expression, and / or reducing or alleviating skin erythema or redness, skin dryness, skin scaling or flaking, and / or skin irritation.

[0011] In some embodiments, the topical skin compositions disclosed herein include any one, any combination, or all of a plankton extract (e.g., saccharide isomers), a reed (Phragmites communis, common reed) extract, a Poria cocos (Poria cocos, smilax glabra root) extract, and a Cucurbita pepo (pumpkin) seed extract. The amount of each component in the composition can vary (e.g., the amount can be as low as 0.000001 weight / weight % to as high as 80 weight / weight %, or any range therebetween). In certain embodiments, a composition comprising any one, any combination, or all of a plankton extract (e.g., saccharide isomers), a reed (common reed) extract, a Poria cocos (Poria cocos, smilax glabra root) extract, and a Cucurbita pepo (pumpkin) seed extract can be formulated into a composition designed to be applied prior to the application of a composition comprising one or more anti-aging compounds (e.g., alpha-hydroxy acids, retinol, etc.). In certain embodiments, any one, any combination or all of a plankton extract (e.g., saccharide isomers), a reed (common reed) extract, a Poria (smilax glabra) extract and a zucchini (pumpkin) seed extract may further comprise one or more anti-aging compounds (e.g., α-hydroxy acids, retinol, etc.). In certain embodiments, any one, any combination or all of a plankton extract (e.g., saccharide isomers), a reed (common reed) extract, a Poria (smilax glabra) extract and a zucchini (pumpkin) seed extract may be formulated into a composition designed to be applied after applying a composition comprising one or more anti-aging compounds (e.g., α-hydroxy acids, retinol, etc.). In certain embodiments, the compositions disclosed herein may further comprise any one or more than one of the ingredients described herein. For example, in certain embodiments, the composition may comprise one or more additional ingredients selected from one or more active ingredients, conditioning agents, moisturizers, pH adjusters, structurants, inorganic salts and / or preservatives. In certain embodiments, the disclosed compositions are used in any one or more than one of the methods described herein.

[0012] In certain embodiments, compositions disclosed herein do not include any one or more than one active ingredient, conditioner, humectant, pH regulator, structurant, inorganic salt and / preservative. In certain embodiments, compositions disclosed herein do not include Argania spinosa oil. In certain embodiments, compositions disclosed herein do not include tetrahexyldecyl ascorbate. In certain embodiments, compositions disclosed herein do not include Acmella oleracea extract. In certain embodiments, compositions disclosed herein do not include Alpinia galanga leaf extract. In certain embodiments, compositions disclosed herein do not include Rosmarinus officinalis leaf extract.

[0013] Also disclosed is the use method of the composition disclosed herein.In certain embodiments, the disclosed composition comprises and is basically composed of or is composed of any one, any combination or all of plankton extract (saccharide isomers), reed (common reed) extract, Poria (Smilax glabra root) extract and / or zucchini (pumpkin) seed extract.In some cases, the method includes applying any one of the compositions disclosed herein topically to skin and / or face and / or eye area in need.In one aspect, any one of the compositions disclosed herein is topically applied, and the composition is left in the application area, removed from the application area after a period of time, and / or directly removed after application.

[0014] In other embodiments, compositions are disclosed, comprising any one, any combination or all of a plankton extract (saccharide isomers), a reed (common reed) extract, a tuckahoe (smilax glabra) extract and / or a squash (pumpkin) seed extract. The composition may be included in a topical skin preparation, an injectable composition, an edible composition or a nutrient. The composition may be in the form of an edible pill or gel capsule or liquid or powder, or spray, or foam, or in an atomized form.

[0015] In a specific aspect, the composition of the present invention is formulated as a topical skin composition. The composition can have a dermatologically acceptable carrier or carrier for compounds, compositions and extracts. The composition can also include moisturizers or wetting agents, surfactants, silicone-containing compounds, UV agents, oily substances and / or other components determined by this specification or components known in the art. The composition can be an emulsion, a cream, a body butter, a facial mask, a scrub, a lotion, a gel, an essence, an emulsion (e.g., oil-in-water, water-in-oil, silicone-in-water, silicone-in-water, water-in-water-in-water, oil-in-water-in-oil, silicone-in-water-in-oil, etc.), a solution (e.g., an aqueous solution or a water-alcohol solution), an anhydrous matrix (e.g., a lipstick or powder), an ointment, an emulsion, a paste, an aerosol, a solid form, an eye gel, etc. The composition can be in powder form (e.g., dry, freeze-dried, granular, etc.). The composition can be formulated for topical skin application at least 1, 2, 3, 4, 5, 6, 7, or more than 7 times per day during use. In some aspects of the invention, the composition can be storage stable or color stable, or both. It is also contemplated that the viscosity of the composition can be selected to achieve the desired result. For example, the viscosity of the composition can range from about 1 cps to well over 1 million cps, or any range or integer therein, depending on the type of composition desired (e.g., 2 cps, 3 cps, 4 cps, 5 cps, 6 cps, 7 cps, 8 cps, 9 cps, 10 cps, 20 cps, 30 cps, 40 cps, 50 cps, 60 cps, 70 cps, 80 cps, 90 cps, 100 cps, 200 cps, 300 cps, 400 cps, 500 cps, 600 cps, 700 cps, 800 cps, 900 cps, 1000 cps, 2000 cps, 3000 cps as measured on a Brookfield viscometer at 25° C. with a TC spindle at 2.5 rpm). , 4000cps, 5000cps, 6000cps, 7000cps, 8000cps, 9000cps, 10000cps, 20000cps, 30000 cps, 40000cps, 50000cps, 60000cps, 70000cps, 80000cps, 90000cps, 100000cps, 2000 00cps, 300000cps, 400000cps, 500000cps, 600000cps, 700000cps, 800000cps, 900000 cps, 1000000cps, 2000000cps, 3000000cps, 4000000cps, 5000000cps, 10000000cps, etc.).

[0016] The compositions of the invention can also be modified to have a desired oxygen radical absorbance capacity (ORAC) value. In certain non-limiting aspects, the compositions or components of the invention identified throughout this specification or extracts thereof can be modified to have at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 90, 10 ... or more than 100,000, or any range of ORAC values ​​therein.

[0017] In non-limiting aspects, the composition can have a pH value of about 6 to about 9. In some aspects, the pH can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14. The composition can include triglycerides. Non-limiting examples include short-chain triglycerides, medium-chain triglycerides and long-chain triglycerides. In some aspects, the triglyceride is a medium-chain triglyceride (e.g., caprylic capric triglyceride). The composition can also include a preservative. Non-limiting examples of preservatives include phenoxyethanol, methylparaben, propylparaben or any mixture thereof. In some embodiments, the composition does not contain parabens.

[0018] The composition of the present invention may have UVA and UVB absorbing properties. The composition may have a sun protection factor (SPF) of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 or more, or any integer obtainable therein. The composition may be a sunscreen lotion, spray or cream.

[0019] The composition of the present invention can also include any one, any combination or all of the following additional ingredients: water, conditioning agent, chelating agent, humectant, pH adjusting agent, inorganic salt, preservative, thickening agent, silicone-containing compound, essential oil, structurant, vitamin, pharmaceutical ingredient or antioxidant, or any combination of these ingredients or the mixture of these ingredients. In some aspects, the composition can be included in at least two, three, four, five, six, seven, eight, nine, ten or more than ten or all of these additional ingredients determined in the aforementioned sentence. The non-limiting examples of these additional ingredients run through this specification sheet, and are incorporated into this section by reference. The amount of these ingredients can be 0.0001% by weight to 99.9% by weight or 0.0001% by volume to 99.9% by volume of the composition, or any integer or scope as disclosed in other parts of this specification sheet, which is incorporated into this section by reference.

[0020] Also contemplated are kits comprising the compositions of the present invention. In certain embodiments, the composition is contained in a container. The container may be a bottle, a dispenser, or a bag. The container may dispense a predetermined amount of the composition. In certain aspects, the composition is dispensed in the form of a spray, mist, a lump, or a liquid. The container may carry a mark on its surface. The mark may be a word, an abbreviation, a picture, or a symbol.

[0021] It is also expected that the composition disclosed in this specification can be used as a wash-free or rinse-off composition. For example, a wash-free composition can be a composition that is topically applied to the skin and retained on the skin for a period of time (e.g., at least 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 20 minutes, or 30 minutes, or at least 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours, or all night or all day). Alternatively, a rinse-off composition can be a product that is applied to the skin and then removed or rinsed (e.g., removed with water) from the skin in a period of time, such as less than 5 minutes, 4 minutes, 3 minutes, 2 minutes, or 1 minute. The example of a rinse-off composition can be a skin cleanser, a shampoo, a conditioner, or a perfumed soap. Examples of leave-on compositions may be skin moisturizers, sunscreens, facial masks, night creams or day creams.

[0022] The following enumerated aspects describe certain inventions disclosed herein.

[0023] Aspect 1 is a method for alleviating skin inflammation in a subject comprising topically applying a cosmetic formulation, wherein the cosmetic formulation comprises at least one transient receptor potential channel vanilloid subtype 1 (TRPV-1) antagonist.

[0024] Aspect 2 is the method of aspect 1, wherein at least one TRPV1 antagonist is a saccharide isomer, a reed extract and a Poria cocos extract and / or a cucurbita pepo (pumpkin) seed extract.

[0025] Aspect 3 is the method of aspect 1 or 2, wherein the cosmetic formulation comprises more than one TRPV-1 antagonist.

[0026] Aspect 4 is the method of any one of aspects 1 to 3, wherein the cosmetic formulation comprises at least 1 weight / volume % of the saccharide isomer.

[0027] Aspect 5 is the method of any one of aspects 1 to 4, wherein the cosmetic formulation comprises at least 5 weight / volume % Cucurbita pepo (pumpkin) seed extract.

[0028] Aspect 6 is the method of any one of aspects 1 to 5, wherein the cosmetic formulation comprises at least 1 weight / volume % of the reed extract and the Poria cocos extract.

[0029] Aspect 7 is the method of any one of aspects 1 to 5, wherein the cosmetic formulation comprises at least 1 weight / volume % of Phragmites australis extract and / or Poria cocos extract.

[0030] Aspect 8 is the method of any one of aspects 1 to 7, wherein the local administration results in inhibition of cellular calcium flux when compared to an appropriate relative control.

[0031] Aspect 9 is the method of any one of aspects 1 to 8, wherein the skin inflammation and / or irritation is caused by agonist activation of TRPV-1.

[0032] Aspect 10 is the method of aspect 9, wherein the agonist of TRPV-1 is a keratolytic agent.

[0033] Aspect 11 is the method of aspect 10, wherein the agonist of TRPV-1 is hydroquinone, resorcinol, trichloroacetic acid, a carboxylic acid, a dicarboxylic acid, a β-hydroxy acid and / or an α-hydroxy acid.

[0034] Aspect 12 is the method of aspect 11, wherein the α-hydroxy acid is citric acid, glycolic acid, lactic acid, malic acid, mandelic acid and / or tartaric acid.

[0035] Aspect 13 is the method of aspect 11, wherein the β-hydroxy acid is salicylic acid, a derivative or metabolite of salicylic acid, β-hydroxybutyric acid, tropic acid and / or trosocaic acid.

[0036] Aspect 14 is the method of aspect 9, wherein the agonist of TRPV-1 is retinol, a retinol precursor, a retinol derivative and / or a retinol metabolite.

[0037] Aspect 15 is the method of aspect 14, wherein the formulation of an ingredient comprising retinol (vitamin A), a retinol precursor, a retinol derivative, or a retinol metabolite comprises at least 0.1% to 1% w / v of the ingredient.

[0038] Aspect 16 is the method of aspect 14 or 15, wherein the formulation comprising retinol and the cosmetic formulation comprising at least one TRPV-1 antagonist are the same composition.

[0039] Aspect 17 is the method of any one of aspects 1 to 16, further comprising administering a formulation comprising at least one pro-inflammatory cytokine inhibitor.

[0040] Aspect 18 is the method of aspect 17, wherein the at least one pro-inflammatory cytokine inhibitor comprises rosemary leaf extract.

[0041] Aspect 19 is the method of aspect 17 or 18, wherein the formulation comprising at least one proinflammatory cytokine inhibitor and the cosmetic formulation comprising at least one TRPV-1 antagonist are the same composition.

[0042] Aspect 20 is the method of aspect 19, wherein the composition comprises retinol.

[0043] Aspect 21 is the method of any one of aspects 17 to 20, wherein the expression of interleukin-6 (IL-6), interleukin-8 (IL-8) and / or tumor necrosis factor alpha (TNFα) is inhibited when compared to an appropriate relative control.

[0044] Aspect 22 is the method of any one of aspects 1 to 21, wherein topical application of the cosmetic formulation reduces erythema, edema, scaling, dryness, burning, stinging, and / or itching.

[0045] Aspect 23 is the method of any one of aspects 1 to 22, wherein application of the cosmetic formulation results in improved skin pigmentation, reduced fine lines and wrinkles, improved evenness of skin tone, reduced photodamage, and / or improved skin firmness when compared to a suitable relative control.

[0046] Aspect 24 is the method of any one of aspects 1 to 23, wherein the cosmetic formulation is formulated for application at least every other day for at least 4 weeks.

[0047] Aspect 25 is the method of aspect 24, wherein the cosmetic formulation is formulated for application at least every other day for at least 8 weeks.

[0048] Aspect 26 is the method of any one of aspects 1 to 24, wherein the cosmetic formulation is formulated as a leave-on product.

[0049] Aspect 27 is the method of any one of aspects 1 to 24, wherein the cosmetic formulation is formulated as a rinse-off product.

[0050] Aspect 28 is a cosmetic formulation comprising at least one TRPV1 antagonist, at least one cytokine inhibitor, and at least 0.1% to 1% w / v retinol, wherein the TRPV1 antagonist is selected from Phragmites australis extract and Poria cocos extract, saccharide isomers, and Cucurbita pepo (pumpkin) seed extract.

[0051] Aspect 29 is the cosmetic formulation of aspect 28, wherein the composition comprises at least 1 weight / volume % of saccharide isomers, at least 5 weight / volume % of cucurbita pepo (pumpkin) seed extract, and / or at least 1 weight / volume % of reed extract and Poria cocos extract.

[0052] Aspect 30 is the cosmetic formulation of aspect 28 or 29, wherein the composition comprises 0.5 weight / volume % retinol.

[0053] It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the invention, and vice versa. In addition, the compositions of the invention can be used to implement the methods of the invention.

[0054] In some embodiments, the composition of the present invention may be pharmaceutically or cosmetically suitable, or may have a pleasant tactile property. "Pharmaceutically suitable", "cosmetically suitable" and / or "comfortable tactile property" describe compositions with specific tactile properties that make the skin feel comfortable (e.g., compositions that are not too watery or too oily, compositions with a silky texture, non-sticky or non-adhesive compositions, etc.). Pharmaceutically or cosmetically suitable may also refer to the creamy or lubricating properties of the composition, or the moisturizing properties of the composition.

[0055] Products comprising the compositions of the present invention are also contemplated. In non-limiting aspects, the product may be a cosmetic. The cosmetic may be those described in other sections of this specification, or those known to those skilled in the art. Non-limiting examples of products include moisturizers, creams, lotions, skin softeners, essences, gels, lotions, body lotions, scrubs, foundations, night creams, lipsticks, cleansers, toners, sunscreens, facial masks, anti-aging products, deodorants, antiperspirants, perfumes, colognes, etc.

[0056] "Topical application" means applying or coating the composition onto the surface of the lips or keratinous tissue. "Topical skin composition" includes compositions suitable for topical application to the skin, lips and / or keratinous tissue. Such compositions are generally dermatologically acceptable because they do not have excessive toxicity, incompatibility, instability, allergic reaction, etc. when applied to the lips, skin and / or keratinous tissue. The topical skin care compositions of the present invention can have a selected viscosity to avoid significant dripping or pooling after application to the lips, skin and / or keratinous tissue.

[0057] "Keratinous tissue" includes the keratin-containing layer that is the outermost protective covering of mammals and includes, but is not limited to, the lips, skin, hair, and nails.

[0058] The term "about" or "approximately" is defined as close to what is understood by those of ordinary skill in the art. In a non-limiting embodiment, the term is defined as within ±10% deviation, preferably within ±5%, more preferably within ±1%, and most preferably within ±0.5%.

[0059] The term "substantially" and variations thereof are defined as being largely, but not necessarily completely, understood by one of ordinary skill in the art, and in one non-limiting embodiment, substantially means within ±10%, within ±5%, within ±1%, or within ±0.5%.

[0060] The terms "inhibit" or "reduce" or any variants of these terms include any measurable reduction or complete inhibition to achieve a desired result. The terms "promote" or "increase" or any variants of these terms include any measurable increase, such as a measurable increase in a protein or molecule (e.g., a matrix protein such as fibronectin, laminin, collagen or elastin or a molecule such as hyaluronic acid) to achieve a desired result.

[0061] The term "effective" as used in the specification and / or claims means sufficient to achieve a desired, expected, or intended result.

[0062] When used in conjunction with the terms "comprises," "comprising," "having," or "containing," or any variations of these terms in the claims and / or description, an element may mean "a," but it also complies with the meanings of "one or more," "at least one," and "one or more than one."

[0063] As used in this specification and claims, the words "comprising," "having," "including," or "containing" are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0064] The compositions and methods of use thereof may "comprise," "consist essentially of," or "consist of" any of the ingredients or steps disclosed in the specification. With respect to the phrase "consist essentially of," the fundamental and novel property of the compositions and methods of the invention is the ability to inhibit TRPV-1 mediated calcium flux as a TRPV-1 antagonist, and / or the ability to act as a demulcent to minimize the activity of a TRPV-1 agonist.

[0065] The phrase "and / or" means "and" or "or". For example, A, B, and / or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.

[0066] Other objects, features and advantages of the present invention will become apparent from the following detailed description. However, it should be understood that the detailed description and examples are given by way of example only when indicating specific embodiments of the present invention. In addition, it is expected that changes and modifications from this detailed description within the spirit and scope of the present invention will be apparent to those skilled in the art. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] The following drawings form part of this specification and are included to further illustrate certain aspects of the present invention. The present invention may be better understood by referring to one or more of these drawings in conjunction with the detailed description of specific embodiments provided herein.

[0068] Figure 1 The inhibition of cellular calcium flux by plankton extracts (e.g., saccharide isomers, also known as (aka) Benoitine) is depicted. In short, Ca 2+ Inward flux was used as a measure of TRPV-1 activity. PC 12 cells (ATCC CRL-1721) were pre-incubated with the corresponding compounds (e.g., plankton extract (test), or capsaicin (positive control)) for 4 h at 37 °C. After pre-incubation, the treatment medium was removed and the cells were incubated with 100 μL of 0.67 μM Fura-2-acetoxymethyl ester ((Fura-2AM), a high affinity intracellular calcium indicator with ratiometric and UV excitability) at 37 °C for 1 h. After incubation, cells were treated with 200 μM capsaicin to induce TRPV-1-mediated Ca 2+ Flux reaction was measured and fluorescence readings were taken every 7 seconds at 360 / 460 nm for 10 minutes. Data were normalized to controls, plotted as shown, and area under the curve (AUC) values ​​were determined (see Table 2). Cells incubated with plankton extracts showed Ca values ​​similar to those incubated with the positive control Capsicum annuum. 2+ Flux curve, where Ca 2+ The flux increased briefly but then decreased rapidly within 2 min.

[0069] Figure 2 The inhibition of cellular calcium flux by each of Phragmites australis (common reed) extract and Poria cocos (Smilax glabra) root extract (e.g., Syricalm™), Cucurbita pekinensis (pumpkin) seed extract (e.g., Ocâline™ PF), and plankton extract (e.g., Saccharide isomers, also known as Benoitine) is depicted. 2+ Inward flux was used as a measure of TRPV-1 activity. PC 12 cells (ATCC CRL-1721) were preincubated with the corresponding test compounds at 37°C for 4 h. After preincubation, the treatment medium was removed and the cells were incubated with 100 μL 0.67 μM Fura-2AM at 37°C for 1 h. After incubation, cells were treated with 200 μM capsaicin to induce TRPV-1-mediated Ca 2+ Flux response was measured and fluorescence readings were taken every 7 seconds at 360 / 460 nm for 10 minutes. Data were normalized to controls, plotted as shown, and AUC values ​​were determined (see Table 2). Cells pre-incubated with extracts of Phragmites australis (common reed) and Poria cocos (smilax glabra) (e.g., Syricalm™), extracts of Cucurbita pekinensis (pumpkin) seeds (e.g., Ocâline™ PF), and plankton extracts (e.g., Saccharide isomers, also known as Benoitine) showed similar Ca2+ to cells incubated with the positive control Capsicum flatulum. 2+ Flux distribution diagram, where Ca 2+ The flux increased but decreased within 2 or 4 minutes, respectively.

[0070] Figure 3 Depicted are histograms documenting the results of a clinical study evaluating the tolerability and efficacy of a composition comprising 0.5% retinol with and without (N=33 individuals) a retinalization regimen (e.g., gradual application of retinol over a period of time). The results showed high tolerability with reduced dermatological assessment of erythema, edema, and dryness when the retinalization regimen was used. Panelists self-assessed fewer moderate sensory discomfort scores and reduced erythema and dryness when the retinalization regimen was used.

[0071] Figure 4 Depicted are histograms documenting the results of a clinical study evaluating clinically measured skin hyperpigmentation, overall fine lines and wrinkles, skin tone uniformity, overall photodamage, and firmness at 4, 8, and 12 weeks of use (N=30 subjects). Implementation of the retinoid regimen provided a cosmetic formulation that provided statistical improvements in each parameter at 4 and 8 weeks (p<0.05). DETAILED DESCRIPTION

[0072] As described above, in certain embodiments, the present invention provides solutions to problems associated with current cosmetic products. In some embodiments, an effective amount of plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (root smilax) extract and / or Cucurbita pekinensis (pumpkin) seed extract is used to inhibit TRPV-1-mediated calcium flux, thereby reducing adverse events, reducing unwanted side effects and / or increasing subject compliance with treatment regimens. Compositions comprising an effective amount of plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (root smilax) extract and / or Cucurbita pekinensis (pumpkin) seed extract can be used to reduce fine lines and wrinkles, even out skin tone, brighten skin tone, increase skin radiance, reduce photodamage, increase elasticity, increase skin barrier function, increase skin firmness, reduce skin sagging, and are superior to those comprising larger amounts of anti-aging compounds (e.g., alpha-hydroxy acids, retinol, etc.).

[0073] In some embodiments, disclosed herein are compositions and methods for inhibiting cellular calcium flux, reducing the release of cell signaling molecules and / or the compliance of an object to a cosmetic treatment regimen. In certain embodiments, the compositions and methods disclosed herein reduce, inhibit and / or substantially eliminate responses to harmful chemical sensory stimuli (e.g., α-hydroxy acids, retinol, etc.). In certain embodiments, this inhibition results in the alleviation of stimulated tissues, the reduction of tissue inflammation, the reduction of tissue redness and / or the reduction of dry skin. In certain embodiments, the compositions and / or methods disclosed herein improve the visual appearance of the skin. In certain embodiments, the compositions and / or methods disclosed herein improve the compliance of an object to a treatment regimen recommended by a clinician and / or manufacturer. In certain embodiments, the compositions and / or methods disclosed herein reduce discomfort, erythema and / or dry skin. In certain aspects, the compositions disclosed herein and methods of use thereof may include, but are not limited to, combinations of ingredients for exfoliating the skin, reducing or eliminating exfoliation irritation, reducing and / or eliminating the appearance of fine lines and / or wrinkles, renewing the skin, increasing skin radiance, soothing the skin, soothing tissue, reducing and / or eliminating inflammation, even skin tone, reducing the appearance of hyperpigmented skin, age spots, liver spots, and / or dark spots, and / or increasing skin smoothness. In certain embodiments, the compositions and / or methods described herein reduce unwanted side effects, reduce adverse events, and / or increase subject compliance with a treatment regimen.

[0074] As shown in Table 2 and Figure 2 As shown, each of the plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (Smilax glabra) root extract, and / or Cucurbita pekinensis (Pumpkin) seed extract showed inhibition of TRPV-1 mediated calcium influx. Figure 3 and Figure 4 As shown, administration of a composition comprising a TRPV-1 inhibitor improves skin characteristics described by subjects and dermatologists, such as skin hyperpigmentation, overall fine lines and wrinkles, skin tone evenness, overall photodamage, skin firmness, erythema, dryness, and / or edema.

[0075] In some embodiments, the composition comprising plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (Smilax glabra root) extract and / or Cucurbita pekinensis (Pumpkin) seed extract can be designed as an emulsion (e.g., oil-in-water, water-in-oil), a gel, a serum, a gel emulsion, a gel serum, a lotion, a mask, a scrub, a wash, a cream, a facial cleanser, a sunscreen, a cleanser, a day cream, a night cream, a body lotion, other skin treatment products, or any combination thereof.

[0076] Some compositions disclosed herein can be applied to the skin or hair and remain on the skin or hair for a period of time (e.g., at least 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 10 minutes, 20 minutes, 30 minutes, or 60 minutes or longer). Thereafter, the composition can be rinsed from the skin or peeled from the skin if desired. Some compositions disclosed herein can be applied to the skin and immediately rinsed from the skin. Some compositions disclosed herein can be applied to the skin and at least partially absorbed by the skin.

[0077] Herein, the inventors show that the addition of one or more than one TRPV-1 antagonist to a cosmetic formulation containing an anti-aging compound (e.g., 0.5% retinol) results in improved compliance with the subject's treatment regimen and reduced erythema, edema, and dryness as assessed by a dermatologist. Retinoids have been widely used to treat photoaging, many of which have shown significant clinical improvements. Although retinoids are the gold standard for treating skin aging, reactions associated with their use, such as burning and peeling, are the main reasons for non-compliance or discontinuation. The retinalization process described herein improves the subject's retinol tolerance by preventing irritation, allowing the subject to comply throughout the study. In addition, the inventors show that less than 6% of the subjects tested for the retinalization regimen reported adverse events during the study, indicating that a combination of TRPV-1 antagonists, nociceptor antagonists, and / or gradual retinalization processes may be a particularly beneficial component in a successful composition and / or method that involves combating signs of aging, particularly by improving tolerance and alleviating irritation caused by anti-aging compounds. In some embodiments, the methods provided herein comprise, consist essentially of, or consist of a TRPV-1 antagonist, a nociceptor antagonist, and / or a combination for combating the gradual retinalization process of signs of aging, particularly by improving tolerability and reducing irritation caused by anti-aging compounds (e.g., irritation caused by retinol, etc.).

[0078] In summary, the studies described herein indicate that the TRPV-1 antagonists described herein can be used to mitigate the irritation associated with the use of high doses of anti-aging compounds (e.g., alpha-hydroxy acids, retinol, etc.) These and other non-limiting aspects of the invention will be described in the following sections.

[0079] A. Active ingredients

[0080] Saccharide isomers are exopolysaccharides that can be synthesized by a microorganism called Vibrio alginolyticus, which belongs to the marine plankton family. In some cases, this ingredient is called a plankton extract. In some cases, this ingredient can be purchased from Barnet Products, which supplies saccharide isomers under the trade name Benoiderm™. In some cases, this ingredient can be purchased under the trade name Benoitine™, also supplied by Barnet Products. It has been determined that an effective amount of saccharide isomers can be used to inhibit TNFα. It is shown herein that an effective amount of saccharide isomers can be used to inhibit TRPV-1 mediated calcium flux.

[0081] Phragmites australis, commonly known as "common reed", but also known as "perennial reed", "ditch reed", "giant reed", "reed grass" or "phragmites". It is a tall, robust perennial herb commonly found in wetlands, swamps, riverbanks and lakesides. Common reed has traditionally been used to improve symptoms associated with indigestion, diabetes, leukemia, breast cancer and insect bites. Common reed is known to contain vitamin A, vitamin C and some B vitamins, but to our knowledge, the potential pathogenic components or mechanisms of action have not been publicly described. It is shown herein that an effective amount of a reed extract can be used to inhibit TRPV-1 mediated calcium flux. In some cases, this ingredient is used in combination with a Poria cocos extract, which is marketed by Chemisches Laboratorium (CLR) under the trade name "SyriCalm"™.

[0082] Poria cocos is a saprophytic fungus of the Polyporaceae family, commonly known as "China root", but also known as "bracket fungus", "Fu-ling", "Hoelen", "Indian Bread", "Tuckahoe", "clerotium cocos", "Wolfiporia extensa", "Wolfiporia cocos", "Daedalea extensa", "Macrohyporia extensa", "Macrohyporia cocos", "Pachyma cocos" or "Medicine of Immortality". Poria cocos grows in and / or on various pine trees, and its sclerotia (vegetative dormant spores) (called "Fuling" in Chinese) have been used as a traditional Chinese medicine for more than two thousand years. Poria cocos has a wide range of biological activities, including anti-tumor, immunomodulatory, anti-inflammatory, antioxidant, anti-aging, anti-hepatitis, anti-diabetic and anti-hemorrhagic fever effects. Poria polysaccharide (PCP) accounts for 84% by weight of all components in the dried sclerotia and is considered to be the main bioactive component. It is shown herein that an effective amount of Poria cocos extract can be used to inhibit TRPV-1 mediated calcium flux. In some cases, this ingredient can be used in combination with a reed extract, which is marketed by Chemisches Laboratorium (CLR) under the trade name "SyriCalm"™.

[0083] Cucurbita pekinensis (Pumpkin) seeds are derived from the Cucurbita pekinensis subsp. Cucurbita pekinensis. Cucurbita pekinensis seed extract has a variety of bioactive compounds with potential nutritional and / or medicinal properties. The beneficial properties of pumpkin seed extract may be due to its strongly dichroic viscous oil. Pumpkin has been used as a treatment option for hypertension and carcinogenic diseases, as an antidiabetic, antibacterial, antioxidant and / or anti-inflammatory agent. It is shown herein that an effective amount of Cucurbita pekinensis seed extract can be used to inhibit TRPV-1 mediated calcium flux. In some cases, such Cucurbita pekinensis (Pumpkin) seed extract is commercially available from Givaudan Active Beauty under the trade name "Ocâline"™.

[0084] The present invention is based on the determination that any one, any combination or all of plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (Smilax glabra) extract and / or Cucurbita pekinensis (Pumpkin) seed extract can be used to inhibit TRPV-1 mediated calcium flux. In some embodiments, inhibiting TRPV-1 mediated calcium flux can relieve skin irritation, reduce skin inflammation, reduce skin redness and / or reduce skin dryness.

[0085] The ingredients disclosed herein can be used in combination with various other cosmetically suitable ingredients, and / or can be used in different product forms to treat various skin diseases. As non-limiting examples, the combination of ingredients can be formulated into an emulsion (e.g., oil-in-water, water-in-oil), a gel, an essence, a gel emulsion, a gel essence, an emulsion, a facial mask, a scrub, a lotion, a cream, or a body lotion.

[0086] The extracts described herein may be extracts prepared by extraction methods and combinations thereof known in the art. Non-limiting examples of extraction methods include the use of liquid-liquid extraction, solid phase extraction, solution extraction, ethyl acetate, ethanol, acetone, oil, supercritical carbon dioxide, heat, pressure, pressure drop extraction, ultrasonic extraction, etc. The extract may be a liquid, a solid, a dried liquid, a resuspended solid, etc.

[0087] B. Amount of ingredients

[0088] It is contemplated that the composition of the present invention may include any amount of the composition discussed in this specification. The composition may also include a combination of any number of additional ingredients described in this specification (e.g., pigment, or additional cosmetic or pharmaceutical ingredients). The concentration of any ingredient may change in the composition. For example, in a non-limiting embodiment, the composition may include, in its final form, substantially consisting of or consisting of, for example, at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.00 23%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051 %, 0.0052%, 0.0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%、0.0425%、0.0450%、0.0475%、0.0500%、0.0525%、0.0550%、0.0575%、0.0600%、0.0625%、0.0650%、0.0675%、0.0700%、0.0725%、0.0750%、0.0775%、0.0800%、0.0825%、0.0850%、0.0875%、0.0900%、0.0925%、0.0950%、0.0975%、0.1000%、0.1250%、0.1500%、0.1750%、0.2000%、0.2250%、0.2500%、0.2750%、0.3000%、0.3250%、0.3500%、0.3750%、0.4000%、0.4250%、0.4500%、0.4750%、0.5000%、0.5250%、0.0550%、0.5750%、0.6000%、0.6250%、0.6500%、0.6750%、0.7000%、0.7250%、0.7500%、0.7750%、0.8000%、0.8250%、0.8500%、0.8750%、0.9000%、0.9250%、0.9500%、0.9750%、1.0%、1.1%、1.2%、1.3%、1.4%、1.5%、1.6%、1.7%、1.8%、1.9%、2.0%、2.1%、2.2%、2.3%、2.4%、2.5%、2.6%、2.7%、2.8%、2.9%、3.0%、3.1%、3.2%、3.3%、3.4%、3.5%、3.6%、3.7%、3.8%、3.9%、4.0%、4.1%、4.2%、4.3%、4.4%、4.5%、4.6%、4.7%、4.8%、4.9%、5.0%、5.1%、5.2%、5.3%、5.4%、5.5%、5.6%、5.7%、5.8%、5.9%、6.0%、6.1%、6.2%、6.3%、6.4%、6.5%、6.6%、6.7%、6.8%、6.9%、7.0%、7.1%、7.2%、7.3%、7.4%、7.5%、7.6%、7.7%、7.8%、7.9%、8.0%、8.1%、8.2%、8.3%、8.4%、8.5%、8.6%、8.7%、8.8%、8.9%、9.0%、9.1%、9.2%、9.3%、9.4%、9.5%、9.6%、9.7%、9.8%、9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% or any range available therein of at least one ingredient mentioned throughout the specification and claims. In non-limiting aspects, the percentages may be calculated by weight or volume of the entire composition. One of ordinary skill in the art will appreciate that the concentrations in a given composition may vary depending on the addition, substitution and / or reduction of ingredients. In certain embodiments, the compositions described herein do not include one or more than one of plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (Smilax glabra root) extract, and / or Cucurbita pekinensis (pumpkin) seed extract. .

[0089] C. Carrier

[0090] The compositions of the present invention may include or be incorporated into all types of carriers and vehicles. The carrier or vehicle may be a pharmaceutically or dermatologically acceptable carrier or vehicle. Non-limiting examples of carriers or vehicles include water, glycerol, alcohol, oil, silicon-containing compounds, silicone compounds, and waxes. Variants and other suitable carriers are apparent to those skilled in the art and are suitable for use in the present invention. In some aspects, the concentration and combination of compounds, ingredients, and reagents are selected in such a way that the combination is chemically compatible and does not form a complex that precipitates from the final product.

[0091] D. Structure

[0092] The compositions of the present invention can be constructed or formulated into various forms. Non-limiting examples include emulsions (e.g., water-in-oil, water-in-oil-in-water, oil-in-water, silicone-in-water, silicone-in-water, oil-in-oil-in-water, silicone-in-water-in-oil emulsions), creams, toners, solutions (aqueous or water-alcoholic), anhydrous bases (e.g., lipsticks and powders), gels, facial masks, scrubs, body lotions, peels, and ointments. Variants and other structures are apparent to the skilled artisan and are suitable for use in the present invention.

[0093] E. Additional Ingredients

[0094] In addition to the combination of ingredients disclosed herein, the composition may also include additional ingredients, such as cosmetic ingredients and pharmaceutical active ingredients. Non-limiting examples of these ingredients are described in the following subsections.

[0095] 1. Cosmetic ingredients

[0096] The CTFA International Cosmetic Ingredient Dictionary and Handbook (2004 and 2008) describes a variety of non-limiting cosmetic ingredients that can be used in the context of the present invention. Examples of these ingredient categories include: fragrances (artificial and natural; for example, gluconic acid, phenoxyethanol, and triethanolamine), dyes and coloring ingredients (for example, Blue 1, Blue 1 Lake, Red 40, titanium dioxide, D&C Blue No. 4, D&C Green No. 5, D&C Orange No. 4, D&C Red No. 17, D&C Red No. 33, D&C Violet No. 2, D&C Yellow No. 10, and D&C Yellow No. 11), flavoring / fragrance agents (for example, Stevia, rebaudiana, sweet leaf) extract and menthol), adsorbents, lubricants, solvents, humectants (including, for example, emollients, humectants, film formers, occlusives and agents that affect the skin's natural moisturizing mechanisms), water repellents, UV absorbers (physical and chemical absorbers, such as para-aminobenzoic acid ("PABA") and corresponding PABA derivatives, titanium dioxide, zinc oxide, etc.), essential oils, vitamins (e.g., A, B, C, D, E and K), trace metals (e.g., zinc, calcium and selenium), anti-irritants (e.g., steroidal and nonsteroidal anti-inflammatory drugs), plant extracts (e.g., Aloe vera, chamomile, cucumber extract, Ginkgo biloba, etc.), biloba), ginseng and rosemary), antimicrobial agents, antioxidants (e.g., BHT and tocopherol), chelating agents (e.g., disodium EDTA and tetrasodium EDTA), preservatives (e.g., methylparaben and propylparaben), pH adjusters (e.g., sodium hydroxide and citric acid), adsorbents (e.g., aluminum starch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrin, talc and zeolite), skin bleaching and whitening agents (e.g., hydroquinone and niacinamide lactate / ester), humectants (e.g., sorbitol, urea, methyl gluceth-20, saccharide isomers and mannitol), exfoliants, water repellents (e.g., magnesium / aluminum hydroxide stearate), skin conditioning agents (e.g., aloe extract, allantoin, bisabolol, ceramides, dimethicone, hyaluronic acid, biosaccharide gum-1, ethylhexylglycerin, pentylene glycol, hydrogenated polydecene, octyldodecyl oleate and dipotassium glycyrrhizate). Non-limiting examples of some of these ingredients are provided in the following subsections.

[0097] a. UV absorbers and / or reflectors

[0098] UV absorbers and / or reflectors that can be used in combination with the compositions of the present invention include chemical and physical sunscreens. Non-limiting examples of chemical sunscreens that can be used include para-aminobenzoic acid (PABA), PABA esters (glyceryl PABA ester, amyl p-dimethylaminobenzoate and octyl p-dimethylaminobenzoate), butyl PABA, ethyl PABA, dihydroxypropanol ethyl PABA, benzophenones (oxybenzone, sulfisobenzone, benzophenone and benzophenone-1 to benzophenone-12), cinnamate salts / esters (octyl methoxycinnamate (cinnamates), isopentyl p-methoxycinnamate, octyl methoxycinnamate, cinoxate, methyl diisopropyl cinnamate, DEA methoxycinnamate, ethyl diisopropyl cinnamate, glyceryl caprylate dimethoxycinnamate and methoxycinnamate Acid ethyl ester), cinnamate, salicylate (homosalate, benzyl salicylate, glycol salicylate, isopropyl benzyl salicylate, etc.), anthranilate / ester, ethyl urocanate, benzoyl salicylate, octyl salicylate, dibenzoylmethane derivatives (such as avobenzone), octocrylene, octyl triazone, galloyl gallate trioleate, aminobenzoic acid glyceryl, 2-hydroxy-1,4-naphthoquinone and dihydroxyacetone, ethylhexyl triazone, dioctyl butyramido triazone, benzylmalonate polysiloxane, terephthalylidene dicamphor sulfonic acid, disodium phenyl dibenzimidazole tetrasulfonate, diethylamino hydroxybenzoyl hexyl benzoate, bis diethylamino hydroxybenzoyl benzoate, dibenzo Azolylphenylethylhexyliminotriazine, drometrizole trisiloxane, methylene bisbenzotriazolyl tetramethylbutylphenol and bisethylhexyloxyphenol methoxyphenyl triazine, 4-methylbenzylidene camphor and isopentyl 4-methoxycinnamate. Non-limiting examples of physical sunscreens include kaolin, talc, petrolatum and metal oxides (such as titanium dioxide and zinc oxide).

[0099] b. Moisturizer

[0100] Non-limiting examples of moisturizers that can be used with the compositions of the present invention include amino acids, chondroitin sulfate, diglycerin, erythritol, fructose, glucose, glycerin, glycerol polymers, ethylene glycol, 1,2,6-hexanetriol, honey, hyaluronic acid, hydrogenated honey, hydrogenated starch hydrolysate, inositol, lactitol, maltitol, maltose, mannitol, natural moisturizing factor, PEG-15 butylene glycol, polyglyceryl sorbitol, salts of pyrrolidone carboxylic acid, PCA potassium, propylene glycol, saccharide isomers, sodium glucuronate, sodium PCA, sorbitol, sucrose, trehalose, urea and xylitol.

[0101] Other examples include acetylated lanolin, acetylated lanolin alcohol, alanine, algae extract, Aloe barbadensis, Aloe barbadensis extract, Aloe barbadensis gel, Althea officinalis extract, Prunus armeniaca kernel oil, arginine, arginine aspartate, Arnica montana extract, aspartic acid, avocado (Persea gratissima) oil, barrier sphingolipids, butanol, beeswax, behenyl alcohol, beta-sitosterol, Betula alba bark extract, Borago officinalis extract, Ruscus aculeatus extract, butylene glycol, Calendula officinalis extract, Calendula oil, Euphorbia cernua cerifera wax, canola oil, caprylic / capric triglyceride, elettaria cardamomum oil, copernicia erifera wax, daucus carota sativa oil, ricinus communis oil, ceramides, ozokerite, ceteareth-5, ceteareth-12, ceteareth-20, cetearyl caprylate, ceteth-20, ceteth-24, cetyl acetate, cetyl caprylate, cetyl palmitate, anthemis nobilis oil, cholesterol, cholesterol esters, cholesterol hydroxystearate, citric acid, sage oil, theobroma cacao butter, coco-caprylate / caprate, coconut oil, collagen, collagen amino acids, zea mays oil mays) oil, fatty acids, decyl oleate, dimethicone copolyol, dimethiconol, dioctyl adipate, dioctyl succinate, dipentaerythrityl hexacaprylate / hexacaprate, DNA, erythritol, ethoxydiglycol, ethyl linoleate, Eucalyptus globulus oil, evening primrose (Oenothera biennis) oil, fatty acids, Geranium punctatummaculatum) oil, glucosamine, glucosyl glutamate / ester, glutamic acid, glyceryl polyether-26, glycerin, glyceryl distearate, glyceryl hydroxystearate, glyceryl laurate, linoleic acid glyceryl, myristic acid glyceryl, oleic acid glyceryl, stearic acid glyceryl, stearic acid glyceryl SE, glycine, glycol stearate, glycol stearate SE, glycosaminoglycan, grape (Vitis vinifera) seed oil, hazelnut (Corylus americana) nut oil, hazelnut (Corylus avellana) nut oil, hexylene glycol, hyaluronic acid, safflower (Carthamus tinctorius) oil, hydrogenated castor oil, hydrogenated coconut glycerides, hydrogenated coconut oil, hydrogenated lanolin, hydrogenated lecithin, hydrogenated palm glycerides, hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenated tallow glycerides, hydrogenated vegetable oil, hydrolyzed collagen, hydrolyzed elastin, hydrolyzed glycosaminoglycan, hydrolyzed keratin, hydrolyzed soy protein, hydroxylated lanolin, hydroxyproline, isocetyl stearate, isocetyl stearyl stearoyl stearate, isodecyl oleate, isopropyl isostearate, isopropyl lanolinate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, isostearamide DEA, isostearic acid, isostearyl lactylate, isostearyl neopentanoate, jasmine (Jasminum officinale) oil, jojoba (Buxus chinensis) oil, giant kelp, stone chestnut (Aleurites moluccana) nut oil, lactamide MEA, lanolin alcohol polyether-16, lanolin alcohol polyether-10 acetate, lanolin, lanolin acid, lanolin alcohol, lanolin oil, lanolin wax, lavender (Lavandula angustifolia) oil, lecithin, lemon (Citrus medica limonum) oil, linoleic acid, linolenic acid, macadamia (Macadamiaternifolia) nut oil, maltitol, matricaria (Chamomilla recutita oil, methyl glucose sesquistearate, methyl silanol PCA esters, mineral oil, mink oil, mortierella fuchsia oil, myristyl lactate, myristyl myristate, myristyl propionate, neopentyl glycol dicaprylate / dicaprate, octyldodecanol, octyldodecanol myristate, octyldodecanol stearoyl stearate, octyl hydroxystearate, octyl palmitate, octyl salicylate, octyl stearate, oleic acid, olive (Olea europaea) oil, orange (Citrus aurantium dulcis) oil, palm (Elaeisguineensis) oil, palmitic acid, pantethine, panthenol, panthenyl ethyl ether, paraffin, PCA, peach (Prunus persica) kernel oil, peanut (Arachis hypogaea) oil, PEG-8C12-18 Esters, PEG-15 Cocoamine, PEG-150 Distearate, PEG-60 Glyceryl Isostearate, PEG-5 Glyceryl Stearate, PEG-30 Glyceryl Stearate, PEG-7 Hydrogenated Castor Oil, PEG-40 Hydrogenated Castor Oil, PEG-60 Hydrogenated Castor Oil, PEG-20 Methyl Glucose Sesquistearate, PEG40 Sorbitan Full Oleate, PEG-5 Soy Sterol, PEG-10 Soy Sterol, PEG-2 Stearate, PEG-8 Stearate, PEG-20 Stearate, PEG-32 Stearate, PEG-40 Stearate, PEG-50 Stearate, PEG-100 Stearate, PEG-150 Stearate, Pentadecanoate, Peppermint (Peppermint, Mentha piperita oil, petrolatum, phospholipids, plankton extract, polyamino acid polysaccharide condensate, polyglyceryl-3 diisostearate, polyquaternium-24, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, potassium myristate, potassium palmitate, propylene glycol, propylene glycol dicaprylate / dicaprate, propylene glycol dicaprylate, propylene glycol dipelargonate, propylene glycol laurate, propylene glycol stearate, propylene glycol stearate SE, PVP, pyridoxine dipalmitate, retinol, retinyl palmitate, rice (Oryza sativa) bran oil, RNA, rosemary (Rosmarinus officinalis) oil, rose oil, safflower (Carthamus tinctorius) oil, sage (Salvia officinalis) oil, sandalwood (Santalum album oil, serine, serum protein, sesame (Sesamum indicum) oil, shea butter (Butyrospermum parkii), silk powder, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodium polyglutamate, soluble collagen, sorbitan laurate, sorbitan oleate, sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol, soybean (Glycine soja) oil, sphingolipids, squalane, squalene, stearamide MEA-stearate, stearic acid, stearoxy polydimethylsiloxane, stearoxy trimethylsilane, stearyl alcohol, stearyl glycyrrhetinate, stearyl heptanoate, stearyl stearate, sunflower (Helianthus annuus) seed oil, sweet almond (Prunus amygdalus dulcis oil, synthetic beeswax, tocopherol, tocopheryl acetate, tocopheryl linoleate, triphein, tridecyl neopentanoate, tridecyl stearate, triethanolamine, tristearin, urea, vegetable oil, water, wax, wheat (Triticum vulgare) germ oil and ylang-ylang (Canangaodorata) oil.

[0102] c. Antioxidants

[0103] Non-limiting examples of antioxidants that can be used with the compositions of the present invention include acetylcysteine, ascorbic acid polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanol pectinate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, cysteine, cysteine ​​hydrochloride, diamylhydroquinone, di-t-butylhydroquinone, dicetylthiodipropionate, dioleyl tocopheryl methylsilanol, disodium ascorbyl sulfate, distearylthiodipropionate, ditridecylthiodipropionate, lauryl gallate, isoascorbic acid, ascorbyl esters, ethyl ferulate, ferulic acid, gallic acid esters, hydroquinone, isooctyl thioglycolate, kojic acid, magnesium ascorbate, magnesium ascorbyl phosphate, methylsilanol Ascorbyl esters, natural plant antioxidants such as green tea or grape seed extracts, nordihydroguaiaretic acid, octyl gallate, phenylthioglycolic acid, potassium ascorbyl tocopheryl phosphate, potassium sulfite, propyl gallate, quinone, rosmarinic acid, sodium ascorbate, sodium bisulfite, sodium isoascorbate, sodium metabisulfite, sodium sulfite, superoxide dismutase, sodium thioglycolate, sorbitan furfural, thiodiglycol, thiodiethylamide, thiodiacetic acid, thioglycolic acid, thiolactic acid, thiosalicylic acid, tocophereth-5, tocophereth-10, tocophereth-12, tocophereth-18, tocophereth-50, tocopherol, tocosorane, tocopheryl acetate, tocopheryl linoleate, tocopheryl nicotinate, tocopheryl succinate, and tris(nonylphenol)phosphite.

[0104] d. Structuring agent

[0105] In other non-limiting aspects, the composition of the present invention may include a structurant. In some aspects, the structurant helps provide rheological characteristics to the composition to contribute to the stability of the composition. In other aspects, the structurant can also act as an emulsifier or surfactant. The non-limiting examples of structurants include stearic acid, palmitic acid, stearyl alcohol, cetyl alcohol, behenyl alcohol, stearic acid, palmitic acid, a polyethylene glycol ether of a stearyl alcohol having an average of about 1 to about 21 ethylene oxide units, a polyethylene glycol ether of a cetyl alcohol having an average of about 1 to about 5 ethylene oxide units, and mixtures thereof.

[0106] e. Emulsifier

[0107] In certain aspects of the invention, the composition does not include an emulsifier. However, in other aspects, the composition may include one or more than one emulsifier. Emulsifiers can reduce interphase surface tension and improve the formulation and stability of the emulsion. Emulsifiers can be nonionic, cationic, anionic and zwitterionic emulsifiers (see U.S. Patent Nos. 5,011,681; 4,421,769; 3,755,560). Non-limiting examples include glycerol esters, propylene glycol esters, fatty acid esters of polyethylene glycol, fatty acid esters of polypropylene glycol, esters of sorbitol, sorbitan anhydride esters, carboxylic acid copolymers, esters and ethers of glucose, ethoxylated esters, ethoxylated alcohols, alkyl phosphates, polyoxyethylene fatty ether phosphates, fatty acid amides, acyl lactylates, fatty acid salts, TEA stearate, oleth-3 phosphate DEA salt, polyethylene glycol 20 sorbitan monolaurate (polysorbate 20), polyethylene glycol 5 soya sterol, steareth-2, steareth-20, steareth-3 phosphate DEA salt ... Ceteth-21, Ceteareth-20, Cetearyl Glucoside, Cetearyl Alcohol, C12-13 Pareth-3, PPG-2 Methyl Glucose Ether Distearate, PPG-5-Ceteth-20, Bis-PEG / PPG-20 / 20 Dimethicone, Ceteth-10, Polysorbate 80, Cetyl Phosphate, Potassium Cetyl Phosphate, Diethanolamine Cetyl Phosphate, Polysorbate 60, Glyceryl Stearate, PEG-100 Stearate, Arachidyl Alcohol, Arachidyl Glucoside, and mixtures thereof.

[0108] f. Silicone-containing compounds

[0109] In non-limiting aspects, silicone-containing compounds include any member of a family of polymeric products in which the backbone of the molecule consists of alternating silicon and oxygen atoms, with side groups attached to the silicon atoms. By varying the length of the -Si-O- chains, the side groups, and the crosslinking, silicones can be synthesized into a wide variety of materials. Their consistency can vary from liquid to gel to solid.

[0110] The silicone-containing compounds that can be used under the environment of the present invention include those described in this specification or known to those of ordinary skill in the art. Non-limiting examples include silicone oils (such as volatile and non-volatile oils), silica gel and silicone solids. In some aspects, the silicone-containing compound includes silicone oils, such as polyorganosiloxanes. The non-limiting examples of polyorganosiloxanes include polydimethylsiloxane, cyclomethicone, polysilicone-11, phenyl trimethicone, trimethylsilyl amino polydimethylsiloxane, stearoxy trimethyl silane or its mixture and other organosiloxane materials of any given ratio, to achieve the desired consistency and application characteristics according to the expected application (for example, applied to specific areas such as skin, hair or eyes). "Volatile silicone oil" includes silicone oils with low heat of vaporization, i.e., usually less than about 50 calories per gram of silicone oil. Non-limiting examples of volatile silicone oils include: cyclomethicone, such as Dow Corning 344 Fluid, Dow Corning 345 Fluid, Dow Corning 244 Fluid and Dow Corning 245 Fluid, Volatile Silicon 7207 (Union Carbide Corp., Danbury, Conn.); low viscosity polydimethylsiloxane, i.e., polydimethylsiloxane having a viscosity of about 50 cst or less (e.g., polydimethylsiloxane, such as Dow Corning 200-0.5 cst fluid). Dow Corning Fluid can be purchased from Dow Corning Corporation in Midland, Michigan. In the third edition of the CTFA Cosmetic Ingredient Dictionary (incorporated by reference), cyclomethicone and polydimethylsiloxane are described as mixtures of cyclic dimethyl polysiloxane compounds and fully methylated linear siloxane polymers terminated with trimethylsiloxy units, respectively. Other non-limiting volatile silicone oils that may be used in the context of the present invention include those available from General Electric Co., Silicone Products Div., Waterford, New York, and SWS Silicones Div., Stauffer Chemical Co., Adrian, Michigan.

[0111] g. Keratolytics

[0112] Exfoliants include ingredients that remove dead skin cells from the outer surface of the skin. These agents can act mechanically, chemically, and / or in other ways. Non-limiting examples of mechanical exfoliants include abrasives such as pumice, silica, cloth, paper, shells, beads, solid crystals, solid polymers, and the like. Non-limiting examples of chemical exfoliants include acid and enzyme exfoliants. Acids that can be used as exfoliants include, but are not limited to, glycolic acid, lactic acid, citric acid, alpha-hydroxy acids, beta-hydroxy acids, and the like. Other exfoliants known to those skilled in the art are also contemplated for use within the scope of the present invention.

[0113] h. Essential oils

[0114] Essential oils include oils from herbs, flowers, trees and other plants. Such oils are usually present as tiny droplets between plant cells and can be extracted by several methods known to those skilled in the art (e.g., steam distillation, enfleurage, i.e., extraction using fats), maceration, solvent extraction or mechanical pressing). When these types of oils are exposed to air, they tend to evaporate (i.e., they are volatile oils). Therefore, although many essential oils are colorless, they are oxidized and become darker in color over time. Essential oils are insoluble in water, but soluble in alcohols, ethers, fixed oils (of plants) and other organic solvents. Typical physical properties of essential oils include a boiling point of about 160°C to 240°C and a density of about 0.759 to about 1.096.

[0115] Essential oils are generally named after the plant they are found in. For example, rose oil or peppermint oil come from the rose or peppermint plant, respectively. Non-limiting examples of essential oils that can be used in the context of the present invention include sesame oil, macadamia oil, tea tree oil, evening primrose oil, Spanish sage oil, Spanish rosemary oil, coriander oil, thyme oil, pimentoberries oil, rose oil, anise oil, balsam oil, bergamot oil, rosewood oil, cedar oil, chamomile oil, sage oil, clary sage oil, clove oil, cypress oil, eucalyptus oil, fennel oil, sea fennel oil, frankincense oil, geranium oil, ginger oil, grapefruit oil, jasmine oil, juniper oil, lavender oil, lemon oil, lemongrass oil, lime oil, mandarin oil,

[0013] The present invention also includes essential oils known to those skilled in the art as neroli oil, orange oil, patchouli oil, pepper oil, black pepper oil, petitgrain oil, pine oil, rose otto oil, rosemary oil, sandalwood oil, spearmint oil, spikenard oil, vetiver oil, wintergreen oil, or ylang-ylang oil. Other essential oils known to those skilled in the art are also contemplated to be useful in the context of the present invention.

[0116] i. Thickener

[0117] Thickeners, including thickeners or gelling agents, include substances that can increase the viscosity of the composition. Thickeners include agents that can increase the viscosity of the composition without substantially changing the efficacy of the active ingredients in the composition. Thickeners can also increase the stability of the compositions of the present invention. In some aspects of the present invention, the thickener includes hydrogenated polyisobutylene, trihydroxystearin, ammonium acryloyldimethyltaurate / VP copolymer, or a mixture thereof.

[0118] Non-limiting examples of additional thickeners that can be used in the context of the present invention include carboxylic acid polymers, cross-linked polyacrylate polymers, polyacrylamide polymers, polysaccharides and gums. Examples of carboxylic acid polymers include cross-linked compounds containing one or more monomers derived from acrylic acid, substituted acrylic acid, and salts and esters of these acrylic acid and substituted acrylic acid, wherein the cross-linking agent contains two or more carbon-carbon double bonds and is derived from a polyol (see U.S. Pat. Nos. 5,087,445; 4,509,949; 2,798,053; CTFA International Cosmetic Ingredient Dictionary, Fourth Edition, 1991, pages 12 and 80). Examples of commercially available carboxylic acid polymers include carbomers, which are homopolymers of acrylic acid cross-linked with allyl ethers of sucrose or pentaerythritol (e.g., CARBOPOL™ 900 series from BF GOODRICH).

[0119] Non-limiting examples of cross-linked polyacrylate polymers include cationic and nonionic polymers. Examples are described in U.S. Pat. Nos. 5,100,660; 4,849,484; 4,835,206; 4,628,078; 4,599,379).

[0120] Non-limiting examples of polyacrylamide polymers, including nonionic polyacrylamide polymers, including substituted branched or unbranched polymers, include polyacrylamide, isoparaffins and laureth-7, multi-block copolymers of acrylamide and substituted acrylamides with acrylic acid and substituted acrylic acid.

[0121] Non-limiting examples of polysaccharides include cellulose, carboxymethyl hydroxyethyl cellulose, cellulose acetate propionate carboxylate, hydroxyethyl cellulose, hydroxyethyl ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl hydroxyethyl cellulose, microcrystalline cellulose, sodium cellulose sulfate, and mixtures thereof. Other examples are alkyl-substituted celluloses, wherein the hydroxyl groups of the cellulose polymer are hydroxyalkylated (preferably hydroxyethylated or hydroxypropylated) to form hydroxyalkylated celluloses, which are then further modified by ether bonds with C10 to C30 straight or branched alkyl groups. Typically these polymers are ethers of C10 to C30 straight or branched alcohols and hydroxyalkyl celluloses. Other useful polysaccharides include scleroglucans, which contain a straight chain of (1-3) linked glucose units with one (1-6) linked glucose per three units.

[0122] Non-limiting examples of gums that can be used in the present invention include gum arabic, agar, algin, alginic acid, ammonium alginate, branched starch, calcium alginate, calcium carrageenan, carnitine, carrageenan, dextrin, gelatin, gellan gum, guar gum, guar hydroxypropyltrimonium chloride, hectorite, hyaluronic acid, hydrated silica, hydroxypropyl chitosan, hydroxypropyl guar gum, karaya gum, kelp, locust bean gum, natto gum, potassium alginate, potassium carrageenan, propylene glycol alginate, sclerotium gum, sodium carboxymethyl dextran, sodium carrageenan, tragacanth gum, xanthan gum, and mixtures thereof.

[0123] j. Preservatives

[0124] Non-limiting examples of preservatives that can be used in the context of the present invention include quaternary ammonium preservatives (e.g., polyquaternium-1 and benzalkonium halides (e.g., benzalkonium chloride ("BAC") and benzalkonium bromide)), parabens (e.g., methylparaben and propylparaben), phenoxyethanol, benzyl alcohol, chlorobutanol, phenol, sorbic acid, thimerosal, or combinations thereof.

[0125] k. Emollients;

[0126] Effective emollients include the following: (a) silicone oils and modifications thereof, such as linear and cyclic polydimethylsiloxanes; amino, alkyl, alkylaryl and aryl silicone oils; (b) fats and oils, including natural fats and oils, such as jojoba oil, soybean oil, sunflower oil, rice bran oil, avocado oil, almond oil, olive oil, sesame oil, peach kernel oil, castor oil, coconut oil, mink oil; cocoa butter; tallow, lard; hardened oils obtained by hydrogenating the above oils; and synthetic mono-, di- and triglycerides. , such as myristic acid glyceryl and 2-ethylhexanoin; (c) waxes, such as carnauba wax, spermaceti, beeswax, lanolin and its derivatives; hydrophobic plant extracts; (e) hydrocarbons, such as liquid paraffin, petrolatum, microcrystalline wax, ozokerite, squalene, pristane and mineral oil; (f) higher fatty acids, such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, lanolin, isostearic acid, arachidonic acid and polyunsaturated fatty acids (PUFAs). A); (g) higher alcohols such as lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, behenyl alcohol, cholesterol and 2-hexanediol; (h) esters such as cetyl octanoate, myristyl lactate, cetyl lactate, isopropyl myristate, myristyl myristate, isopropyl palmitate, isopropyl adipate, butyl stearate, decyl oleate, cholesterol isostearate, glyceryl monostearate, glyceryl distearate, glyceryl tristearate, alkyl lactates, alkyl citrates and (i) essential oils and extracts thereof, for example, mint, jasmine, camphor, white cypress, bitter orange peel, ryu, turpentine, cinnamon, bergamot, mandarin orange, calamus, pine, lavender, laurel, clove, cypress, eucalyptus, lemon, carambola, thyme, peppermint, rose, sage, sesame, ginger, basil, juniper, lemongrass, rosemary, rosewood, avocado, grape, grape seed, myrrh, cucumber, watercress, calendula, elderflower, geranium, linden flower (linden (j) lipids, such as cholesterol, ceramides, sucrose esters and pseudoceramides as described in European Patent Specification No. 556,957; (k) vitamins, minerals and skin nutrients, such as vitamin A, vitamin E and vitamin K; vitamin alkyl esters, including vitamin C alkyl esters; magnesium, calcium and milk; (l) sunscreens, such as octyl methoxycinnamate (Parsol MCX) and butyl methoxybenzoyl methane (Parsol MCX). 1789); (l) phospholipids; (m) polyhydric alcohols, such as glycerol and propylene glycol; and polyols, such as polyethylene glycol;(n) anti-aging compounds, such as alpha-hydroxy acids, beta-hydroxy acids; and (o) mixtures of any of the foregoing components, etc.;

[0127] l. Thickener

[0128] Examples of suitable tackifiers include, but are not limited to, aliphatic hydrocarbon resins, aromatic modified aliphatic hydrocarbon resins, hydrogenated polycyclopentadiene resins, polycyclopentadiene resins, gum rosin, gum rosin esters, wood rosin, wood rosin esters, tall oil rosin, tall oil rosin esters, polyterpenes, aromatic modified polyterpenes, terpene phenols, aromatic modified hydrogenated polycyclopentadiene resins, hydrogenated aliphatic resins, hydrogenated aliphatic aromatic resins, hydrogenated terpenes and modified terpenes, hydrogenated rosin acids, hydrogenated rosin esters, polyisoprene, partially or fully hydrogenated polyisoprene, polybutene diene, partially or fully hydrogenated polybutene diene, etc. As demonstrated by some of the cited examples, the tackifier may be fully or partially hydrogenated. The tackifier may also be non-polar. (By non-polar it is meant that the tackifier is substantially free of monomers having polar groups. Preferably, polar groups are not present, however, if they are present, they are preferably present in an amount up to about 5 weight percent, preferably up to about 2 weight percent, more preferably up to about 0.5 weight percent.).

[0129] m. Colorant

[0130] The composition of the present invention may also include at least one cosmetically acceptable colorant, such as a pigment or dye. Examples of suitable pigments include, but are not limited to, inorganic pigments, organic pigments, lakes, pearlescent pigments, iridescent or optically variable pigments and mixtures thereof. Pigments are understood to be inorganic or organic, white or colored particles. Within the scope of the present invention, the pigment may optionally be surface treated, but is not limited to treatments such as silicones, perfluorochemicals, lecithin and amino acids.

[0131] n. Surfactant

[0132] Surfactants useful as the surfactant component in the compositions of the present invention include nonionic, anionic, cationic and amphoteric (zwitterionic) surfactants, and may be used in combination with one another.

[0133] o. pH adjuster

[0134] pH adjusters include inorganic and organic acids and bases, particularly ammonia, citric acid, phosphoric acid, acetic acid, sodium hydroxide, lactic acid, levulinic acid, glycolic acid, tartaric acid, malic acid, pyrrolidonecarboxylic acid (PCA), succinic acid, citric acid, glutamic acid, 2-amino-2-methyl-1-propanol (AMP) and triethanolamine (TEA).

[0135] p. Reducing agent

[0136] Suitable reducing agents include, but are not limited to, thiourea, thiosulfates (e.g., sodium salt), sulfites, bisulfites, metabisulfites, borohydrides, and hypophosphites, ascorbic acid and its salts, esters, and derivatives (e.g., ascorbyl palmitate and ascorbic acid polypeptides), and tocopherol and its salts, esters, and derivatives (e.g., tocopherol acetate).

[0137] q. Fragrance

[0138] The compositions disclosed herein may optionally contain a fragrance. Examples of possible fragrances include natural oils or naturally derived materials, and synthetic fragrances such as hydrocarbons, alcohols, aldehydes, ketones, esters, lactones, ethers, nitriles, and polyfunctional compounds. Non-limiting examples of natural oils include the following: basil (Ocimum basilicum) oil, bay (Pimento acris) oil, bee balm (Monarda didyma) oil, bergamot (Citrus aurantium bergamia) oil, cardamom (Elettaria cardamomum) oil, cedar (Cedrus atlantica) oil, chamomile (Anthemis nobilis) oil, cinnamon (Cinnamomum cassia) oil, lemongrass (Cymbopogon nardus) oil, sage (Salvia sclarea) oil, clove (Eugenia caryophyllus) oil, clover (Eufenia caryophyllus) oil, sedge oil (Cyperus esculentus oil), cypress (Cupressus sempervirens) oil, lemon eucalyptus (Eucalyptus globulus) oil. citriodora oil, geranium maculatum oil, ginger oil, grapefruit oil, hazelnut oil, jasmine oil, juniper oil, juniper tar, juniper virginiana oil, kiwi juice, lavender oil, lavender oil, lavender juice, lemon oil, lemongrass oil, citron oil, citronella ... schoenanthus) oil, lemon (Citrus aurantifolia) oil, lime (Tilia corodia) oil, lime (TiliaTilia corodia juice, Guangxi mandarin orange (Citrus nobilis) oil, nutmeg (Myristica fragrans) oil, orange (Citrus aurantium dulcis) flower oil, orange (Citrus aurantium dulcis) oil, orange (Citrus aurantium dulcis) juice, patchouli (Pogostemoncablin) oil, peppermint (Menthe piperita) oil, peppermint (Menthe peperita) juice, rosemary (Rosmarinus officinalis) oil, rose oil, Damascena rose (Rosa damascena) extract, wild rose (Rosamultiflora) extract, rosewood (Aniba rosaeodora) extract, sage (Salvia officinalis) oil, sandalwood (Santalum album) oil, spearmint (Menthe viridis) oil, tea tree (Melaleuca alternifolia, Melaleuca alternifolia oil and Cananga odorata oil. Some non-limiting examples of synthetic hydrocarbon fragrances include caryophyllene, β-farnesene, limonene, α-pinene and β-pinene. Some non-limiting examples of synthetic alcohol fragrances include bacdanol, citronellol, linalool, phenylethyl alcohol and α-terpineol (R═H). Some non-limiting examples of synthetic aldehyde fragrances include 2-methyl undecene aldehyde, citral, hexyl cinnamaldehyde, isocyclohexyl cinnamaldehyde, lily of the valley aldehyde and 10-undecene aldehyde. Some non-limiting examples of synthetic ketone fragrances include cashmeranone, α-ionone, ambroxanone E, acetyl dipoly-unusual pentenes (koavone), musk ketone and tuna musk. Some non-limiting examples of synthetic ester fragrances include benzyl acetate, 4-tert-butylcyclohexyl acetate (cis and trans), cedryl acetate, cyclohexyl acetate, isobornyl acetate, and alpha-pinene acetate (R = acetyl). Some non-limiting examples of synthetic lactone fragrances include coumarin, jasmine lactone, muskalactone, and peach aldehyde. Some non-limiting examples of synthetic ether fragrances include ambergris, anthers, and galaxolide. Some non-limiting examples of synthetic nitrile fragrances include cinnamonitrile and gernonitrile. Finally, some non-limiting examples of synthetic multifunctional compound fragrances include amyl salicylate, isoeugenol, hedione, heliotropine, lyral, and vanillin. ,

[0139] r. Foaming agent

[0140] The foaming agent includes, for example, sodium lauryl sulfate, sodium lauroyl sarcosinate, sodium alkyl sulfosuccinate, sodium coconut oil fatty acid monoglycerol sulfonate, sodium α-olefin sulfonate, N-acylamino acid salts such as N-fatty acyl glutamate, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, maltitol fatty acid ester, sucrose fatty acid ester, polyglycerol fatty acid ester, fatty acid diethanolamide, polyoxyethylene sorbitan monostearate, polyoxyethylene hydrogenated castor oil and polyoxyethylene fatty acid ester. These foaming agents can be used alone or in combination of two or more.

[0141] s. Tanning agent

[0142] Suitable tanning agents include, but are not limited to, α-hydroxyaldehydes and ketones, glyceraldehyde and related alcohol aldehydes, various indoles, imidazoles and their derivatives, and various approved colorants. Other suitable tanning agents include, but are not limited to, methylglyoxal, glyceraldehyde, erythrulose, alloxan, 2,3-dihydroxysuccinaldehyde, 2,3-dimethoxysuccinaldehyde, 2-amino-3-hydroxysuccinaldehyde, and 2-benzylamino-3-hydroxysuccinaldehyde.

[0143] t. Astringent

[0144] Suitable astringents include, but are not limited to, aluminum citrate, aluminum lactate, birch extract, coffee extract, evening primrose extract triclocarbon, grape extract, henna extract, ivy extract, lemon extract, witch hazel extract, ammonium and potassium alum, aluminum tripolyphosphate, aluminum glycinate and aluminum phenol sulfate, allantoin chlorohydrate aluminum (Alcloxa), allantoin hydroxyaluminum (Aldioxa), aluminum stearate, aluminum sulfate and aluminum citrate, sodium aluminum phosphate, sodium alum, sodium aluminum chlorohydroxy lactate, calcium lactate, calcium chloride, calcium sulfate hydrate, sodium aluminum lactate, zinc acetate, zinc chloride, zinc sulfate, zinc lactate, zinc zeolite, zinc phenol sulfonate, and combinations thereof. Extracts refer to the entire fruit, legume and / or plant, or selected components of such fruit, legume and / or plant.

[0145] u. Fungicide

[0146] Suitable antiseptics include, but are not limited to, methyl, ethyl, propyl or butyl paraben, phenoxyethanol, o-phenylphenol, dehydroacetic acid or its salts, p-cresol, m-cresol, o-chloro-m-xylenol, peppermint oil, echinacea, American bloodroot, pepper, tea tree oil, wild bergamot, chaparral, stinging nettle, laurel, myrrh, rhatany bark, toothache tree, calendula, chamomile, mupirocin, neomycin sulfate, bacitracin, polymyxin B, levofloxacin, tetracyclines (chlortetracycline hydrochloride, oxytetracycline hydrochloride and tetracycline hydrochloride), clindamycin phosphate, gentamicin sulfate, benzalkonium chloride, benzethonium chloride, hexylresorcinol, methylbenzethonium chloride, phenol, quaternary ammonium compounds, triclocarban, triclosan and tea tree oil.

[0147] v. Deodorants and antiperspirants

[0148] Suitable antiperspirants and deodorants include, but are not limited to, zinc salts such as zinc sulfate and zinc chloride, glycinates such as aluminum zirconium glycinate, aluminum hydrochloride, aluminum zirconium tetrachloroglycinate, zinc carbonate, o-phenylphenol and quaternary ammonium compounds such as dimethylbenzylammonium chloride and hexamethonium chloride.

[0149] w. Whitening agent

[0150] Examples of skin whitening agents include, but are not limited to, hydroquinone, kojic acid, licorice and / or its derivatives, ascorbic acid and / or its derivatives, arbutin, bearberry extract, glycyrrhiza glabra and its derivatives, chlorella extract, perilla extract, coconut extract and / or other depigmenting agents.

[0151] x. Antimicrobial agents

[0152] Examples of antimicrobial agents include, but are not limited to, triclosan, 3,4,4'-trichlorocarbanilide (triclocarban); 3,4,4'-trifluoromethyl-4,4'-dichlorocarbanilide (chlorofluorocarbanilide); 5-chloro-2-methyl-4-isothiazolin-3-one; iodopropynyl butylcarbamate; 8-hydroxyquinoline; 8-hydroxyquinoline citrate; 8-hydroxyquinoline sulfate; 4-chloro-3,5-xylenol (chloroxylenol); 2-bromo-2-nitropropane-1,3-diol; diazolidinyl urea; butoconazole; nystatin; terconazole; nitrofurantoin; phenazopyridine; acyclovir; clotrimazole ; Chloroxylenol; Chlorhexidine; Miconazole; Terconazole; Butylparaben; Ethylparaben; Methylparaben; Methylchloroisothiazoline; Methylisothiazoline; A mixture of 1,3-bis(hydroxymethyl)-5,5-dimethylhydantoin and 3-iodo-2-propynylbutylcarbamate; Hydroxyquinoline; EDTA; Tetrasodium EDTA; Parabens; Alkylpyridinium compounds; Coco-phosphatidyl PG-diammonium chloride; Chlorhexidine gluconate; Chlorhexidine digluconate; Chlorhexidine acetate; Chlorhexidine isethionate; Chlorhexidine hydrochloride; Benzalkonium chloride; Benzethonium chloride; Polyhexamethylene biguanide; and mixtures thereof.

[0153] 2. Drug ingredients

[0154] Pharmaceutically active agents are also contemplated as useful in the compositions of the present invention. Non-limiting examples of pharmaceutical actives include anti-acne agents, agents for treating rosacea, analgesics, anesthetics, anorectal agents, antihistamines, anti-inflammatory agents including nonsteroidal anti-inflammatory drugs, antibiotics, antifungals, antivirals, antimicrobials, anticancer actives, scabicides, pedicicides, anti-tumor agents, antiperspirants, antipruritics, antipsoriatic agents, antiseborrheic agents, biologically active proteins and peptides, burn treatments, cautery agents, depigmenting agents, depilatories, diaper rash treatments, enzymes, hair growth stimulants, hair growth retardants including DFMO and its salts and analogs, hemostats, keratolytics, oral ulcer treatments, cold sore treatments, dental and periodontal treatments, photosensitizing actives, skin protectants / barrier agents, steroids including hormones and corticosteroids, sunburn treatments, sunscreens, transdermal actives, nasal actives, vaginal actives, wart treatments, wound treatments, wound healing agents, and the like.

[0155] F. Kit

[0156] Also contemplated are kits for use in certain aspects of the present invention. For example, the composition of the present invention may be included in a kit. The kit may include a container. The container may include a bottle, a metal tube, a laminated tube, a plastic tube, a dispenser, a high pressure container, a barrier container, a package, a compartment, a lipstick container, a compacted container, a cosmetic tray or other type of container capable of storing a cosmetic composition, such as an injection or blow molded plastic container, wherein a dispersion or composition or a desired bottle, dispenser or package is stored. The kit and / or container may include a label on its surface. For example, the label may be a word, a phrase, an abbreviation, a picture or a symbol.

[0157] The container can distribute the composition of a predetermined amount. In other embodiments, a container (such as metal, laminate or plastic tube) can be squeezed to distribute the composition of the desired amount. The composition can be distributed as a spray, an aerosol, a liquid, a fluid or a semisolid. The container can have a spray mechanism, a pump mechanism or an extrusion mechanism. The test kit can also include instructions for using the test kit components and using any composition contained in the container. Instructions can include explanations of how to apply, use and preserve the composition. Example

[0158] The following examples are used to illustrate the preferred embodiments of the present invention. It will be appreciated by those skilled in the art that the disclosed technology in the following examples can represent the technology found by the inventors to play a good role in the practice of the present invention, and therefore it can be considered that it constitutes the preferred embodiment for its practice. However, according to the present disclosure, it will be appreciated by those skilled in the art that many changes can be made in the disclosed specific embodiments without departing from the spirit and scope of the present invention, and still obtain the same or similar results.

[0159] All compositions and / or methods disclosed and claimed herein can be made and implemented according to the present disclosure without undue experimentation. Although the compositions and methods of the present invention have been described in accordance with preferred embodiments, it is obvious to those skilled in the art that changes can be implemented in the compositions and / or methods and in the steps or the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the present invention. More specifically, it is obvious that specific reagents related to both chemical and physiological aspects can replace the reagents described herein while achieving the same or similar results. All such similar substitutions and changes that are obvious to those skilled in the art are considered to be within the spirit, scope and concept of the present invention as defined by the appended claims.

[0160] Example 1

[0161] (Materials used)

[0162] The active ingredients in Table 1 were used to obtain the in vitro and / or clinical data described below and / or in the associated figures.

[0163] Table 1

[0164] .

[0165] Example 2

[0166] (In vitro activity)

[0167] It has been determined that any of the plankton extract (saccharide isomers), Phragmites australis (common reed) extract, Poria cocos (Smilax glabra) root extract, and / or Cucurbita pekinensis (Pumpkin) seed extract can be used to inhibit TRPV-1 mediated calcium flux. The results are summarized in Table 2, and the methods used to determine the properties of the ingredients are provided below.

[0168] The ability of the component samples to inhibit calcium flux generation in PC 12 cells (ATCC CRL-1721) was analyzed. PC 12 cells were maintained and passaged according to the manufacturer's recommended instructions. For the experiments, 1×10 5 Cells / well were seeded in a 96-well black flat-bottom plate. Fura-2-acetoxymethyl ester (Fura-2AM) calcium detection assay kit (Cayman Chemical #14591) was used to detect calcium in the treated cells. Capsaicin (Sigma # M202) was used to initiate the reaction, and capsaicin (Sigma # C191) was used as a positive control. In brief, Ca 2+ Inward flux is used as a measure of TRPV-1 activity. PC 12 cells (ATCC CRL-1721) were pre-incubated with the corresponding test compounds at 37°C for 4 hours. After pre-incubation, the treatment medium was removed and the cells were incubated with 100 μL 0.67 μM Fura-2AM at 37°C for 1 hour. After incubation, the cells were treated with 200 μM capsaicin to induce TRPV-1-mediated Ca2+ flux response, and fluorescence readings were read every 7 seconds at 360 / 460 nm for 10 minutes. After the test period, the buffer was removed and 100 μL of diluted MTS reagent was added to each well. The cells were incubated at 37°C under 5% CO2 for 15 to 30 minutes, and the plate readings were read at 490 nm. Cell viability was assessed before analyzing calcium flux.

[0169] Fluorescence test data were normalized to controls to generate area under the curve (AUC) values. For each control and test, the fluorescence arbitrary units (AU) were normalized to the controls at time points 0 min and 0.5 min to 9 min. The normalized data were plotted on an XY graph using GraphPad Prism software to generate a calcium flux curve with time on the X axis and normalized AU on the Y axis. The AUC for the untreated control was then calculated, and the net AUC for each test and control condition was determined. The data for the test conditions are expressed as a percentage of the positive control (capsazepine 400µM) (results are shown in Table 2).

[0170] Cells pre-incubated with plankton extracts (e.g., Saccharide isomers, also known as Benoitine), Phragmites australis (common reed) extracts, and Poria cocos (Smilax glabra) root extracts (e.g., Syricalm™), and Cucurbita pekinensis (pumpkin) seed extracts (e.g., Ocâline™ PF), respectively, showed similar Ca2+ levels to cells incubated with the positive control Capsicum annuum. 2+ Flux curve, where Ca 2+ The flux increased but decreased within 2 or 4 minutes, respectively.

[0171] Table 2

[0172] Results of analysis of capsaicin-induced TRPV-1-mediated calcium flux

[0173] .

[0174] Example 3

[0175] (Clinical Research)

[0176] In clinical studies, it has been determined that a subject's compliance with a treatment regimen can be improved by including one or more than one TRPV-1 antagonist in the treatment regimen. Figure 3 and Figure 4 In conjunction with the results shown, a summary of the clinical study methods and results used to determine the properties of the ingredients is provided below.

[0177] The inventors conducted a series of clinical studies to evaluate the tolerability and efficacy of a composition containing 0.5% retinol in order to develop a retinolization regimen to improve tolerability in the general population. The study design included a preliminary study in which panelists used the retinol formulation every other night to establish a baseline sensory profile. Figure 3 and Figure 4Based on the results shown, and in consultation with dermatologists, a gradual retinolization regimen was developed to maximize tolerability and maintain efficacy. Study endpoints included dermatologist assessment of erythema, edema, flaking, and dryness, and panelists' self-assessment of burning, stinging, and itching. Additional clinical studies were conducted to assess skin pigmentation, overall fine lines and wrinkles, skin tone uniformity, overall photodamage, and firmness as clinical indicators. A summary description of the clinical studies related to the above assessments can be found in Example 4 below.

[0178] Example 4

[0179] (Additional determination)

[0180] Assays that can be used to determine the efficacy of any ingredient, or any combination of ingredients or compositions disclosed throughout this specification and claims comprising a combination of ingredients can be determined by methods known to those of ordinary skill in the art. The following are non-limiting assays that can be used in the context of the present invention. It should be appreciated that other testing procedures can be used, including, for example, objective and subjective procedures.

[0181] B16 Pigmentation Assay: Melanogenesis is the process by which melanocytes produce melanin, a naturally occurring pigment that gives color to the skin, hair, and eyes. Inhibition of melanogenesis is beneficial in preventing skin darkening and reducing dark spots associated with aging. This bioassay uses an immortalized mouse melanoma cell line, B16-F1 melanocytes (ATCC), to analyze the effects of compounds on melanin production. The endpoints of this analysis are spectrophotometric measurements of melanin production and cell activity. B16-F1 melanocytes can be cultured in standard DMEM growth medium with 10% fetal bovine serum (MEDIATECH) at 37°C in 10% CO2 and then treated with any of the active ingredients, combinations of ingredients, or compositions having said combinations disclosed in this specification. After incubation, cell activity is quantified by measuring melanin secretion by absorption at 405nm.

[0182] Collagen stimulation assay: Collagen is an extracellular matrix protein that is very important for the structure of the skin. Increased collagen synthesis helps improve skin firmness and elasticity. This bioassay can be used to detect the effect of any of the active ingredients, combinations of ingredients, or compositions having said combinations disclosed in this specification on the production of procollagen peptides (precursors of collagen) by human epidermal fibroblasts. The endpoint of this test is a spectrophotometric measurement reflecting the presence of procollagen peptides and cell activity. The analysis uses a quantitative sandwich enzyme immunoassay technique, so monoclonal antibodies specific for procollagen peptides are pre-coated on microplates. Standards and samples can be pipetted into the wells, and all procollagen peptides present are bound by the immobilized antibodies. After washing away all unbound material, an enzyme-linked polyclonal antibody specific for procollagen peptides is added to the wells. After rinsing to remove all unbound antibody-enzyme reactants, a substrate solution can be added to the wells, and the color development is proportional to the amount of procollagen peptide bound in the initial step, and is detected at 450nm using a microplate reader. The color development can be stopped and the intensity of the color measured.

[0183] To generate samples and controls, subconfluent normal human mature epidermal fibroblasts (Cascade Biologics) cultured in standard DMEM growth medium with 10% fetal bovine serum (MEDIATECH) at 37° C. in 10% CO 2 can be treated with each of the combinations of components disclosed in the specification or compositions having said combinations for 3 days. After incubation, the cell culture medium can be collected, and the amount of procollagen peptide secretion can be quantified using TAKARA's sandwich enzyme-linked immunosorbent assay (ELISA) (#MK101).

[0184] Elastin Stimulation Assay: Elastin is a connective tissue protein that helps the skin regain its shape after being stretched or contracted. Elastin is also an important load-bearing protein used where mechanical energy needs to be stored. Elastin is made by joining many soluble tropoelastin molecules in a reaction catalyzed by the enzyme lysyl oxidase. Elastin secretion and elastin fibers can be monitored in cultured human fibroblasts by staining them with an immunofluorescent antibody against elastin.

[0185] Laminin and Fibronectin Stimulation Assay: Laminin and Fibronectin are the major proteins in the dermal-epidermal junction (DEJ), also known as the basement membrane. The DEJ is located between the dermis and the epidermis and interlocks to form finger-like projections, called reticular ridges. Epidermal cells receive their nutrients from blood vessels in the dermis. The reticular ridges increase the surface area of ​​the epidermis exposed to these blood vessels and required nutrients. The DEJ provides adhesion of the two tissue compartments and controls the structural integrity of the skin. Laminin and Fibronectin are two glycoproteins located in the DEJ. Laminin and Fibronectin are considered to be adhesives that glue cells together and are secreted by dermal fibroblasts to help promote intracellular and intercellular adhesion of the epidermis called DEJ. The secretion of laminin and fibronectin can be monitored by quantifying laminin and fibronectin in the cell supernatant of cultured human fibroblasts treated for 3 days with medium containing or not containing 1.0% final concentration of the test component. After incubation, the content of laminin and fibronectin can be measured in an enzyme-linked immunosorbent assay (ELISA) using immunofluorescent antibodies against laminin and antibodies against fibronectin. The measurement of cellular metabolic activity is normalized as determined by the bioconversion of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazole (MTS).

[0186] Tumor Necrosis Factor Alpha (TNF-α) Assay: TNF-α, the prototype ligand of the TNF superfamily, is a pleiotropic cytokine that plays a major role in inflammation. Increased expression of it is associated with upregulated proinflammatory activity. This bioassay can be used to detect the effect of any of the active ingredients, combinations of ingredients, or compositions having said combinations disclosed in this specification on the production of TNF-α by human epidermal keratinocytes. The endpoint of the assay may be a spectrophotometric measurement reflecting the presence of TNF-α and cell activity. The assay uses a quantitative sandwich enzyme immunoassay technique, so a monoclonal antibody specific for TNF-α has been pre-coated on a microplate. Standards and samples can be pipetted into the wells, and any TNF-α present is bound by the immobilized antibody. After washing away all unbound material, an enzyme-linked polyclonal antibody specific for TNF-α can be added to the wells. After washing to remove all unbound antibody-enzyme reactants, a substrate solution can be added to the wells, and color development is proportional to the amount of TNF-α bound in the initial step, detected at 450 nm using a microplate reader. The color development can be stopped and the intensity of the color can be measured. Subconfluent normal adult keratinocytes (Cascade Biologics) cultured in EPILIFETM standard growth medium (Cascade Biologics) at 37°C in 5% CO2 can be treated with phorbol 12-myristic acid 13-acetate (PMA, 10ng / mL, SIGMA CHEMICAL, #P1585-1MG) and any of the active ingredients disclosed in the specification, ingredient combinations, or compositions with the combination for 6 hours. PMA has been shown to cause a significant increase in TNF-α secretion, which peaks 6 hours after treatment. After cultivation, cell culture media can be collected and the amount of TNF-α secretion can be quantified using a sandwich enzyme-linked immunosorbent assay (ELISA) from R&D Systems (#DTA00C).

[0187] Antioxidant (AO) assay: An in vitro bioassay that measures the total antioxidant capacity of any of the ingredients, combinations of ingredients, or compositions having said combinations disclosed herein. The assay relies on the ability of antioxidants in a sample to inhibit the ABTS ® (2,2'-Azobis-[3-ethylbenzothiazoline sulfonate]) is oxidized to ABTS by metmyoglobin ®+. The antioxidant system of living organisms contains enzymes such as superoxide dismutase, catalase and glutathione peroxidase; macromolecules such as albumin, ceruloplasmin and ferritin; and a large number of small molecules, including ascorbic acid, α-tocopherol, β-carotene, reduced glutathione, uric acid and bilirubin. The total number of endogenous food-derived antioxidants represents the total antioxidant capacity of the extracellular fluid. The collaboration of all different antioxidants provides stronger protection against the attack of reactive oxygen or nitrogen free radicals than any single compound alone. Therefore, the total antioxidant capacity can give more relevant biological information than that obtained by measuring individual components because it takes into account the cumulative effect of all antioxidants present in plasma and body fluids. The ability of antioxidants in the sample to prevent ABTS oxidation was compared with Trolox, a water-soluble tocopherol analog, and quantified as the molar equivalent of Trolox. As an in vitro bioassay for measuring the total antioxidant capacity of any of the ingredients, combinations of ingredients, or compositions having said combinations disclosed in this specification, the antioxidant capacity kit # 709001 from CAYMAN CHEMICAL (Ann Arbor, Michigan USA) can be used. The protocol can be performed according to the manufacturer's recommendations.

[0188] ORAC assay: The oxygen radical absorption (or absorption) capacity (ORAC) of any of the active ingredients disclosed in this specification, the combination of active ingredients, or the composition containing the combination can also be tested by measuring the antioxidant activity of these ingredients or compositions. Antioxidant activity indicates the ability to reduce oxidants (oxidants). This analysis quantifies the time and degree required to inhibit the action of oxidants, such as oxygen free radicals, which are known to cause damage to cells (such as skin cells). The ORAC value of any of the active ingredients disclosed in this specification, the combination of active ingredients, or the composition containing the combination can be determined by methods known to those of ordinary skill in the art (see US Publication Nos. 2004 / 0109905 and 2005 / 0163880; and commercially available kits such as Zen-Bio ORAC Antioxidant Assay Kit (#AOX-2)). The Zen-Bio ORAC Antioxidant Assay Kit measures the loss of fluorescein fluorescence over time due to the formation of hydroperoxide-free radicals by the decomposition of AAPH (2,2'-azobis-2-methylpropylimidazole dihydrochloride). Trolox is a water-soluble vitamin E analog used as a positive control that inhibits fluorescein decay in a dose-dependent manner.

[0189] Mushroom tyrosinase activity assay: In mammalian cells, tyrosinase catalyzes two steps in the biosynthesis of melanin from tyrosine (and from dopachrome polymerization). Tyrosinase is located in melanocytes and produces melanin (aromatic quinone compound) that gives skin, hair and eyes color. Mushroom tyrosinase (SIGMA) can be cultured with its substrate L-Dopa (FISHER) in the presence or absence of any of the active ingredients disclosed in this specification, the combination of ingredients, or the combination having said combination. Pigment formation can be assessed by colorimetric plate reading at 490nm. The percentage inhibition of mushroom tyrosinase activity can be calculated by comparison with an untreated control to determine the ability of the test ingredient or its combination to inhibit the activity of the purified enzyme. The inhibitory effect of the test extract was compared with the inhibitory effect of kojic acid (SIGMA).

[0190] Matrix metalloproteinase 3 and matrix metalloproteinase 9 activity (MMP3; MMP9) assay: In vitro matrix metalloproteinase (MMP) inhibition assay. MMPs are extracellular proteases that play a role in many normal and disease states by virtue of their broad substrate specificity. MMP3 substrates include collagen, fibronectin, and laminin; whereas MMP9 substrates include collagen VII, fibronectin, and laminin. Using colorimetric drug discovery kits for MMP3 (AK-400) and MMP-9 (AK-410) from BioMol International, the assay is designed to measure the protease activity of MMPs using a sulfur-containing polypeptide as a chromogenic substrate (Ac-PLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5)5,6. The peptide bond of the MMP cleavage site is replaced by a thioester bond in the sulfur-containing polypeptide. This bond is hydrolyzed by MMP to generate a thiol group, which reacts with DTNB [5,5'-dithiobis(2-nitrobenzoic acid), Ellman's reagent] to generate 2-nitro-5-thiobenzoic acid, which can be detected by its absorbance at 412 nm (ε = 13600 M-1cm at pH 6.0 and above 7). -1 ). Any of the active ingredients, ingredient combinations disclosed in this specification, or compositions containing the combinations may be tested.

[0191] Matrix Metalloproteinase 1 Enzyme Activity (MMP1) Assay: In vitro matrix metalloproteinase (MMP) inhibition assay. MMPs are extracellular proteases that play a role in many normal and disease states by virtue of their broad substrate specificity. MMP1 substrates include collagen IV. The Molecular Probes ENZ / CHEK Gelatinase / Collagenase Assay Kit (#E12055) utilizes a fluorescent gelatin substrate to detect MMP1 protease activity. After proteolytic cleavage, bright green fluorescence is displayed and can be observed using a fluorescent microplate reader to measure enzyme activity.

[0192] The ENZ / CHEK Gelatinase / Collagenase Assay Kit (#E12055) from Invitrogen is designed as an in vitro assay for measuring MMP1 enzyme activity. Any of the active ingredients, combinations of ingredients disclosed in this specification, or compositions containing said combinations can be tested. The analysis relies on the ability of the pure MMP1 enzyme to degrade a fluorescent gelatin substrate. Once the substrate is specifically cleaved by MMP1, bright green fluorescence is displayed and can be observed using a fluorescent microplate reader. The test material is cultured in the presence or absence of pure enzyme and substrate to determine its protease inhibition ability.

[0193] Cyclooxygenase (COX) assay: In vitro cyclooxygenase-1 and cyclooxygenase-2 (COX-1, COX-2) inhibition assay. COX is a bifunctional enzyme with both cyclooxygenase and peroxidase activity. The cyclooxygenase activity converts arachidonic acid to hydroperoxide endoperoxides (prostaglandin G2; PGG2), and the peroxidase component reduces the endoperoxides (prostaglandin H2; PGH2) to the corresponding alcohols, prostaglandins, and coagulation The COX inhibition screening assay measures the peroxidase component of cyclooxygenase. Peroxidase activity is measured by colorimetry by monitoring the appearance of oxidized N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD). The inhibition screening assay includes both COX-1 and COX-2 enzymes to screen isozyme-specific inhibitors. The colorimetric COX (sheep) inhibitor screening test (#760111, CAYMAN CHEMICAL) can be used to analyze the influence of each active ingredient disclosed in the specification, any combination of ingredients, or a composition with the combination on the activity of purified cyclooxygenase (COX-1 or COX-2). According to the manufacturer's instructions, the purified enzyme, heme, and test extract can be mixed in an assay buffer and incubated at room temperature for 15 minutes with shaking. After cultivation, arachidonic acid and a colorimetric substrate can be added to start the reaction. The color change can be evaluated by a colorimetric plate measured at 590nm. The percent inhibition of COX-1 or COX-2 activity can be calculated by comparison with untreated controls to determine the ability of the test extract to inhibit the activity of the purified enzyme.

[0194] Lipoxygenase (LO) assay: In vitro lipoxygenase inhibition assay. LO is a non-heme, iron-containing dioxygenase that catalyzes the addition of molecular oxygen to fatty acids. Linoleate and arachidonate are the main substrates of LO in plants and animals. Arachidonic acid can then be converted to hydroxyeicosatetraenoic (HETE) acid derivatives, which are subsequently converted to leukotrienes, potent inflammatory mediators. This assay provides an accurate and convenient method for screening lipoxygenase inhibitors by measuring the hydroperoxides produced by lipoxygenases (5-LO, 12-LO, or 15-LO) cultured with arachidonic acid. The ability of each of the active ingredients disclosed in this specification, a combination of ingredients, or any of the compositions having the combination to inhibit enzyme activity can be determined using a colorimetric LO inhibitor screening kit (#760700, CAYMAN CHEMICAL). Purified 15-lipoxygenase and the test components can be mixed in assay buffer and incubated with shaking at room temperature for 10 minutes. After incubation, arachidonic acid can be added to initiate the reaction, and the mixture can be incubated for another 10 minutes at room temperature. A colorimetric substrate can be added to terminate catalysis, and the color change can be assessed by a fluorescent plate measured at 490 nm. The percent inhibition of lipoxygenase activity can be assessed compared to an untreated control to determine the ability of each of the active ingredients disclosed in this specification, a combination of ingredients, or any of the compositions having said combinations to inhibit the activity of the purified enzyme.

[0195] Elastase assay: The ENZCHEK® Elastase assay (Kit # E-12056) of Molecular PROBES (Eugene, Oregon USA) can be used as an in vitro enzyme inhibition assay for measuring the inhibition of elastase activity of any of the active ingredients disclosed in this specification, the combination of ingredients, or the composition having the combination. The ENZCHEK kit contains soluble bovine neck ligament elastin, which can be labeled with a dye to quench the fluorescence of the conjugate. Non-fluorescent substrates can be digested by elastase or other proteases to produce highly fluorescent fragments. The resulting fluorescence enhancement can be monitored with a fluorescent microplate reader. The digestion products from the elastin substrate have an absorption maximum at about 505nm and a fluorescence emission maximum at about 515nm. When using the ENZCHEK Elastase Assay Kit to screen for elastase inhibitors, the peptide, N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone, can be used as a selective, common elastase inhibitor.

[0196] Ceramide production: Ceramide in cells or tissue samples can be labeled with mouse monoclonal antibody anti-ceramide (Enzo Life Sciences, reference ALX-804-196 clone MID15B4) diluted to 1 / 50 for 2 hours at room temperature using the amplification system biotin / streptavidin. Video microscopy can be performed to visualize ceramide (pink staining).

[0197] Oil control assay: An analysis for measuring the reduction of sebum secretion in sebaceous glands and / or the reduction of sebum production in sebaceous glands can be analyzed by using standard techniques known to those of ordinary skill in the art. In a specific example, the forehead can be used. Each of the active ingredients disclosed in this specification, the combination of ingredients, or any of the compositions having the combination can be applied once a day or twice a day to a portion of the forehead for a fixed period of days (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days or more than 14 days), while the rest of the forehead is not treated with the composition. After a fixed period of days has expired, sebum secretion can be analyzed by applying fine blotting paper to the treated and untreated forehead skin. This is accomplished by first removing all sebum from the treated and untreated areas with wet and dry cloths. The blotting paper is then applied to the treated and untreated forehead areas, and a rubber band can be placed around the forehead to gently press the blotting paper onto the skin. After 2 hours, the blotter can be removed, allowed to dry, and then transilluminated. Darker blotters correspond to more sebum secretion (or lighter blotters correspond to reduced sebum secretion.

[0198] Erythema Assay: An assay to measure the reduction in skin redness can be evaluated using a MINOLTA colorimeter. Skin erythema can be induced by applying a 0.2% sodium lauryl sulfate solution to the subject's forearm. The area is protected with an occlusive patch for 24 hours. After 24 hours, the patch is removed and the irritation-induced redness can be evaluated using the a* value of a MINOLTA colorimeter. The a* value measures the change in skin color in the red area. Immediately following the reading, the area is treated with any of the active ingredients, combinations of active ingredients, or compositions containing said combinations disclosed in this specification. Repeated measurements can be made periodically to determine the ability of the formulation to reduce redness and irritation.

[0199] Determination of skin moisture / hydration: The benefits of skin moisture / hydration can be measured by using impedance measurements performed with a Nova DermalPhase Meter. Impedance meters measure changes in skin moisture content. The outer layer of the skin has different electrical properties. When the skin is dry, it conducts electricity poorly. When it becomes more hydrated, increased conductivity is produced. Therefore, changes in skin impedance (related to conductivity) can be used to evaluate changes in skin hydration. The device can be calibrated for each test day according to the instrument instructions. Temperature and relative humidity can also be marked. The subject can be evaluated as follows: before measurement, it can be balanced in a room with a determined humidity (e.g., 30% to 50%) and temperature (e.g., 68°C to 72°C). Three independent impedance measurements are performed on each side of the face, and they are recorded and averaged. The impedance meter can use the T5 setting, which averages the impedance values ​​applied to the face every five seconds. Changes can be reported with statistical variance and significance. Each of the active ingredients disclosed in this specification, any one of the ingredient combinations, or a composition containing the combination can be tested according to this method.

[0200] Determination of Skin Clarity and Reduction of Freckles and Age Spots: Skin clarity and reduction of freckles and age spots can be evaluated using a Minolta colorimeter. Changes in skin color can be evaluated using the a* value of a Minolta colorimeter to determine potential irritation due to product treatment. The a* value measures changes in skin color in the red area. This is used to determine whether each of the active ingredients disclosed in this specification, any of the combinations of ingredients, or a composition containing the combination reduces irritation. Measurements can be made on each side of the face and averaged as values ​​for the left and right sides of the face. Skin clarity can also be measured using a Minolta Meter. The measurement is a combination of the a*, b, and L values ​​of the Minolta Meter and is related to the brightness of the skin, and corresponds very well to the smoothness and hydration of the skin. Skin determinations are performed as above. In one non-limiting aspect, skin clarity can be described as L / C, where C is chromaticity and is defined as (a / C). 2 + b 2 ) 1 / 2 .

[0201] Skin dryness, surface wrinkles, skin smoothness, and skin color measurement: Skin dryness, surface wrinkles, skin smoothness, and skin color can be assessed using a clinical scoring technique. For example, a clinical score for skin dryness can be determined using a five-point standard Kligman scale: (0) skin is soft and moist; (1) skin appears normal with no visible dryness; (2) skin feels slightly dry to the touch with no visible flaking; (3) skin feels dry, tough, and has a whitish appearance with some scaling; and (4) skin feels very dry, rough, and has a whitish appearance with scaling. The assessment can be performed independently by two clinicians and averaged.

[0202] Skin color clinical score measurement: Skin color was clinically scored using a 10-point analog numeric scale: (10) Smooth uniform skin, pink-brown color. No dark, red, or scaly patches when examined with a handheld magnifier. The microtexture of the skin feels very uniform; (7) Uniform skin color without magnification. No scaly areas, but macules due to hyperpigmentation or erythema. No macules larger than 1 cm in diameter; (4) Skin macules and uneven texture are easily noticed. Few scales. Skin that feels rough to the touch in some areas; and (1) Uneven skin coloration and texture. Multiple areas of scales and macules, hypopigmented, red, or black spots. Large areas of uneven coloration greater than 1 cm in diameter. The assessments were performed independently by two clinicians and averaged.

[0203] Clinical Score Measurement of Skin Smoothness: The clinical score of skin smoothness can be analyzed using a ten-point analog numerical scale: (10) smooth, the skin is moist and shiny, and there is no resistance when a finger is passed over the surface; (7) somewhat smooth, little resistance; (4) rough, visibly changing, and there is friction when rubbed; and (1) rough, flaky, uneven surface. The assessment was performed independently by two clinicians and the average was taken.

[0204] Skin smoothness and wrinkle reduction measurements using the method disclosed in Packman et al. (1978): Skin smoothness and wrinkle reduction can also be measured visually using the method disclosed in Packman et al. (1978). For example, the depth, shallowness, and total number of surface face lines (SFL) for each subject can be carefully scored and recorded at each visit. A numerical score is obtained by multiplying the number factor by the depth / width / length factor. Scores are obtained for the eye area and mouth area (left and right) and added together as a total wrinkle score.

[0205] Skin firmness measurement using Hargens Ballistometer: Skin firmness can be measured using the Hargens Ballistometer, a device that assesses skin elasticity and firmness by dropping a small object on the skin and recording the first two rebound peaks. Ballistometry is a small, lightweight probe that uses a relatively blunt probe (4 square millimeters - contact area). The probe gently penetrates into the skin, resulting in measurements that rely on the properties of the outer layers of the skin, which include the stratum corneum and outer epidermis and part of the dermis.

[0206] Skin Softness / Flexibility Measurement Using Gas Bearing Electrodynamometer: Skin softness / flexibility can be assessed using the Gas Bearing Electrodynamometer, an instrument that measures the pressure / tension properties of the skin. The viscoelasticity of the skin is related to skin moisturization. Measurements of specific sites in the cheek area can be achieved by attaching the probe to the skin surface with double-sided tape. A force of approximately 3.5 gm is applied parallel to the skin surface, accurately measuring the displacement of the skin. The flexibility of the skin can then be calculated and expressed as DSR (dynamic spring rate in gm / mm).

[0207] Determination of the appearance of lines and wrinkles using replicas: The appearance of lines and wrinkles on the skin can be evaluated using replicas, which are impressions of the skin surface. Materials such as silicone rubber can be used. The replicas can be analyzed by image analysis. Changes in line and wrinkle visibility can be objectively quantified by forming the subject's face using a silicon replica and analyzing the replica image with a computer image analysis system. Replicas can be obtained from the eye area and the neck area and photographed with a digital camera at a low lighting incident angle. The digital image can be analyzed with an image processing program and the area of ​​the replica covered by wrinkles and fine lines can be determined.

[0208] Skin surface profile determination using the profilometer / stylus method: The surface profile of the skin can be measured using the profilometer / stylus method. This involves flashing or dragging a stylus across the surface of a replica. The vertical displacement of the stylus can be recorded in a computer via a distance sensor, and after scanning a certain distance of the replica, an analysis of the skin profile can be generated as a two-dimensional surface. This scan can be repeated any number of times along a fixed axis to produce a simulated 3-D image of the skin. Ten random sections of the replica can be obtained using the stylus technique and combined to produce an average. Values ​​of interest include Ra, which is the arithmetic mean of all roughness (height) values ​​calculated by integrating the profile height relative to the mean profile height. Rt, which is the maximum vertical distance between the highest peak and the lowest valley, and Rz, which is the average peak amplitude minus the average peak height. Values ​​are given as values ​​calibrated in mm. The equipment should be standardized before each use by scanning a metal standard of known value. The Ra value can be calculated by the following formula: R a = normalized roughness; lm = lateral (scan) length; and y = absolute value of profile position relative to the average profile height (x-axis).

[0209] MELANODERM™ Assay: In other non-limiting aspects, the efficacy of each of the active ingredients disclosed in this specification, any combination of ingredients, or compositions having such combinations can be evaluated by using a skin analog such as MELANODERM™. Melanocytes, one type of cell in the skin analog, are specifically contaminated when exposed to L-dihydroxyphenylalanine (L-DOPA), a precursor of melanin. The skin analog, MELANODERM™, can be treated as follows: using various matrices containing each of the active ingredients, any combination of ingredients, or compositions containing the combinations disclosed in the specification or using the matrix alone as a control. Alternatively, an untreated skin analog sample can be used as a control.

[0210] Production of filaggrin: Changes in the production of filaggrin in keratinocytes caused by each of the active ingredients disclosed in this specification, any one of the ingredient combinations, or a composition containing the combination can be measured. Filaggrin is a precursor of the natural moisturizing factor (NMF) in the skin. An increase in NMF increases the moisture content of the skin. Using a bioassay that analyzes the concentration of filaggrin in keratinocyte lysates, the production of filaggrin in treated and untreated keratinocytes can be determined. A non-limiting example of a bioassay that can be used to quantify the production of intermediate filament-related proteins is the PROTEINSIMPLESIMON protein blotting protocol. For each sample, normal human epidermal keratinocytes (NHEKs) were grown in EPI-200-MATTEK EPI Life growth medium (M-EP-500-CA) containing calcium from LifeTechnologies. Before treatment, NHEKs were incubated overnight at 37°C in a growth medium with 5% CO2. The NHEKs are then incubated in growth medium containing 1% of the test compound / extract or no compound / extract (negative control) for 24 hours to 36 hours. The NHEKs can then be washed, collected, and stored on ice or in a cooler place until lysed on ice using lysis buffer and sonication. The protein concentration of the sample can be determined and used for normalization of the samples. The lysate can be stored at -80°C until used for quantitative analysis.

[0211] PROTEINSIMPLE ® The SIMON™ Western Blot Bioassay uses a quantitative Western blot immunoassay technique to quantify filaggrin in test samples using a filaggrin-specific antibody. Cell samples are lysed and normalized for protein concentration. Normalized samples and molecular weight standards can then be loaded and run on a denaturing protein separation gel using capillary electrophoresis. The proteins in the gel are immobilized and immunoprobed using a primary antibody specific for filaggrin. The immobilized proteins can then be immunoprobed with an enzyme-linked detection antibody that is bound to the primary antibody. A chemiluminescent substrate solution can then be added to the immobilized proteins, allowing chemiluminescence development proportional to the amount of filaggrin bound in the fixative. The development of chemiluminescence is stopped at a specific time, and the intensity of the chemiluminescent signal can be measured and compared to positive and negative controls.

[0212] Production of occlusin: Changes in the production of occlusin in keratinocytes caused by each of the active ingredients disclosed in this specification, any one of the ingredient combinations, or a composition containing the combination can be measured. Occlusin is a protein that is essential for the formation of tight junctions and the moisture barrier function of the skin. A non-limiting example of how to determine the production of occlusin in treated and untreated keratinocytes is a bioassay using an analysis of the concentration of occlusin in keratinocyte lysates. Bioassays can be performed using the PROTEINSIMPLE® SIMON™ protein blotting protocol. For samples, adult epidermal keratinocytes (HEKa) from Life Technologies (C-005-5C) can be grown in EPILIFE™ growth medium containing calcium from Life Technologies (M-EP-500-CA) and keratinocyte growth supplement (HKGS) from Life Technologies (S-101-5) at 37°C and 5% CO2 for 24 hours. The HEKa are then incubated for 24 to 48 hours in growth medium containing the test compound / extract, no compound / extract as a negative control, or 1 mM CaCl2 as a positive control. The HEKa are then washed, collected, and stored on ice or colder until lysed on ice using lysis buffer and sonication. The protein concentration of the sample can be determined and used for normalization of the samples. The lysate is stored at -80°C until used in the bioassay.

[0213] The PROTEINSIMPLE® SIMON™ Western Blot Bioassay utilizes a quantitative Western blot immunoassay technique that uses an antibody specific for occludin to quantitatively detect occludin in a test sample. Cell samples are lysed and the protein concentration is normalized. The normalized samples and molecular weight standards are then loaded and run on a denaturing protein separation gel using capillary electrophoresis. The proteins in the gel are then immobilized and immunoprobed using a primary antibody against occludin. The immobilized proteins are immunoprobed using an enzyme-linked detection antibody that is bound to the primary antibody. A chemiluminescent substrate solution is then added to the immobilized proteins, allowing chemiluminescence development proportional to the amount of occludin bound in the fixative. The development of chemiluminescence can be stopped at a specific time, and the intensity of the chemiluminescent signal can be measured and compared to positive and negative controls.

[0214] Corneocyte monolayer permeability: Changes in the permeability of the corneocyte monolayer caused by each of the active ingredients disclosed in this specification, any one of the ingredient combinations, or a composition containing the combination can be measured. Corneocyte monolayer permeability is a measure of skin barrier integrity. As a non-limiting example, the in vitro vascular permeability test of MILLIPORE (ECM642) can be used to determine the corneocyte monolayer permeability in treated and untreated corneocytes. This test analyzes the adsorption, transport and permeability of endothelial cells. In short, adult epidermal corneocytes from Life Technologies (C-005-5C) can be seeded onto a porous collagen-coated membrane in a collection well. The corneocytes are then cultured in EPILIFE™ growth medium containing calcium from LIFE TECHNOLOGIES (M-EP-500-CA) and corneocyte growth supplement (HKGS) from LIFETECHNOLOGIES (S-101-5) at 37°C and 5% CO2 for 24 hours. This culture time allows the cells to form a monolayer and seal the membrane pores. The culture medium is then replaced with fresh culture medium containing (test sample) or without (untreated control) the test compound / extract, and the corneocytes are cultured for another 48 hours at 37°C and 5% CO2. In order to determine the permeability of the corneocyte monolayer after culture with / without the test compound / extract, the culture medium is replaced with fresh culture medium containing high molecular weight fluorescein isothiocyanate (FITC)-dextran, and the corneocytes are cultured for 4 hours at 37°C and 5% CO2. During the 4-hour incubation, FITC can pass through the corneocyte monolayer and the porous membrane and enter the collection well at a rate proportional to the permeability of the monolayer. After incubation for 4 hours, the cell viability and FITC content in the collection well can be determined. For FITC content, the medium in the collection well, when excited at 520nm, measures the fluorescence of the medium at 480nm (Em). The percent permeability and percent change compared to the untreated control can be determined by the following formulas: Permeability = ((average Ex / Em of test samples) / average Ex / Em of untreated controls)*100; Percent change = Permeability of test samples – Permeability of untreated controls.

[0215] Production of hyaluronic acid: Changes in the production of hyaluronic acid in human dermal fibroblasts due to each of the active ingredients disclosed in this specification, any one of the combinations of ingredients, or a composition containing the combination can be measured. HA is a polysaccharide that participates in the stabilization of the matrix structure and provides turgor pressure to tissues and cells.

[0216] As a non-limiting example, the HA production in treated and untreated adult dermal fibroblasts (HDFa) can be determined using the hyaluronic acid DuoSetELISA kit of R&D Systems (DY3614). In this analysis, in order to produce samples, sub-fusion HDFa cells from Cascade Biologics (C-13-5C) were incubated at 37°C and 10% CO2 in starvation medium (0.15% fetal bovine serum and 1% penicillin-streptomycin solution in Dulbecco's modified Eagle medium) for 72 hours before treatment. The cells were then incubated with fresh starvation medium containing test compounds, positive controls (phorbol 12-myristic acid 13-acetate (P1585) from SIGMA-ALDRICH and platelet-derived growth factor (P3201) from SIGMA-ALDRICH) or without additives for 24 hours. The culture medium was then collected and frozen at -80°C until used for ELISA determination.

[0217] In short, the ELISA assay uses a quantitative sandwich enzyme immunoassay technique, whereby a capture antibody specific for HA can be pre-coated onto a microplate. Standards and culture media from treated and untreated cells are pipetted into the microplate wells to allow any HA present to be bound by the fixed antibody. After all unbound material is washed away, an enzyme-linked detection antibody specific for HA is added to the wells. After washing to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells to allow color development proportional to the amount of HA bound in the initial step. Color development is stopped at a specific time, and the color intensity at 450nm is measured using a microplate reader.

[0218] As another non-limiting example, human skin explants can be cultured in a survival explant culture medium in a humidified atmosphere at 37°C, supplemented with 5% CO2. The treatment of the explants can be carried out by topical application of sample products (n = 3) at day 0, day 2, day 3, day 6, day 8 and day 9. Control explants (n = 3) do not receive any treatment except renewing the survival explant culture medium. At day 3, day 6 and day 8, a survival culture medium of half volume can be renewed. At day 9, three explants under each condition can be taken out and cut in half. Half of the explants are fixed in buffered formalin, and the other half are frozen at -80°C.

[0219] After fixing in common Bouin for 48 hours and in formalin for 24 hours, the sample can be dried and immersed in paraffin using the automatic tissue processor Leica TP 1020. 5 micron sections can be made with a microtome (Minot type LEICA RM2125) and mounted on SUPERFROST™ histology slides. Microscopic observation can be carried out by using the optical microscope of a LEICAORTHOPLAN or LEICA DM LB microscope. The image can be taken with an Olympus DP72 camera and CELL^D software. General morphology can be checked on paraffin sections stained with the tricolor Goldner variant of Masson. The staining of hyaluronic acid can be diluted to 1 / 100 1 hour at room temperature with anti-hyaluronic acid biotinylated protein (HABP) (SEIKAGAKU ref 400763-1A), carried out with amplification system biotin / streptavidin (carrier, VECTASTAIN PK-7200).

[0220] Inhibition of hyaluronidase activity: Changes in the activity of hyaluronidase due to each of the active ingredients disclosed in this specification, any one of the combinations of ingredients, or a composition containing the combination can be measured. Hyaluronidase is an enzyme that degrades hyaluronic acid. HA is a polysaccharide that participates in the stability of matrix structure and provides turgor pressure for tissues and cells. As a non-limiting example, an in vitro protocol modified from SIGMA-ALDRICH protocol # EC 3.2.1.35 can be used to determine hyaluronidase activity. In short, 1-S type hyaluronidase (H3506) from SIGMA-ALDRICH is added to a microplate reaction well containing a test compound or a control. Tannic acid can be used as a positive control inhibitor, the control enzyme can not be added to the test compound, and the well containing the test compound or the positive control but not containing the hyaluronidase can be used as a background negative control. Before adding the substrate (HA), the well was incubated at 37°C for 10 minutes. Substrate was added and the reactants were incubated at 37°C for 45 minutes. A portion of each reaction solution is then transferred to a solution of sodium acetate and acetic acid, pH 3.75 and gently mixed to stop that portion of the reaction (stopped well). After adding a portion of the reaction solution to the stopped well, the stopped well and the reaction well should both contain the same volume of solution. Both the reaction well and the stopped well are incubated at room temperature for 10 minutes. The absorbance of the reaction well and the stopped well is then measured at 600nm. The inhibition can be calculated using the following formula: Inhibitor (or control) activity = (absorbance of inhibitor stop well at 600nm – absorbance of inhibitor reaction well at 600nm); Initial activity = absorbance of control enzyme at 600nm; Percent inhibition = [(initial activity / inhibitor activity)*100]-100.

[0221] Peroxisome proliferator-activated receptor gamma (PPAR-γ) activity: Changes in PPAR-γ activity caused by each active ingredient disclosed in the specification, any combination of ingredients, or a composition having the combination can be measured. PPAR-γ is a key receptor for sebum production. As a non-limiting example, the activity of PPAR-γ can be determined using a bioassay that analyzes the ability of a test compound or composition to inhibit ligand binding. Briefly, a fluorescent small molecule pan-PPAR ligand, FLUORMONE™ Pan-PPAR Green, available from Life Technologies (PV4894), can be used to determine whether a test compound or composition is able to inhibit the binding of the ligand to PPAR-γ. Sample wells include PPAR-γ and a fluorescent ligand and: a test compound or composition (test); a reference inhibitor, rosiglitazone (positive control); or no test compound (negative control). The wells are incubated for a set period of time to allow the ligand an opportunity to bind to PPAR-γ. The fluorescence polarization of each sample well can then be measured and compared to the negative control well to determine the inhibition rate of the test compound or composition.

[0222] Cytokine Array: Human epidermal glial cells were cultured to 70% to 80% confluence. The medium in the plate was aspirated and 0.025% trypsin / EDTA was added. When the cells became rounded, the culture dish was tapped to release the cells. The cells containing trypsin / EDTA were removed from the culture dish and balanced. The cells were centrifuged for 5 minutes. To form a cell pellet. The supernatant was aspirated. The resulting pellet was resuspended in EPILIFE™ medium (Cascade Biologics). The cells were seeded into 6-well plates at approximately 10% to 20% confluence. When the cells become approximately 80% confluent, the medium is aspirated and 1.0 mL of EPILIFE™ is added to duplicate wells along with phorbol 13-myristate 12-acetate (“PMA”), an inflammatory inducer for transplantation, and a test composition dilution (i.e., 1.0% (100 μL of 100× stock) and 0.1% (10 μL of 100× stock) test composition diluted into a final volume of 1 mL of EPILIFE™ growth medium). The medium is gently vortexed to ensure adequate mixing. In addition, 1.0 mL of EPILIFE™ is added to the control wells, with or without the addition of PMA. Following preparation, the plates are then incubated at 37±1°C and 5.0 ± 1% CO2 for approximately 5 hours. After the 5 hour incubation, all medium is collected in conical tubes and frozen at -70°C

[0223] For analysis, 16-grid hybridization boxes were attached to three 16-grid FAST sheets arranged in duplicate with 16-grid anti-cytokine antibodies plus experimental controls (WHATMAN BIOSCIENCES), and the sheets were placed in FAST frames (4 sheets per frame) for processing. The array was blocked for 15 minutes. 70mL S&S protein array blocking buffer (WHATMANSCHLEICHER and SCHEULL) was used at room temperature. The blocking buffer was removed and 70mL of each supernatant sample was added to each array. The array was incubated at room temperature for 3 hours with gentle stirring. The array was washed 3 times with TBS-T. The array was processed with 70mL antibody mixture, which contained a biotinylated antibody corresponding to each array capture antibody. The array was incubated at room temperature for 1 hour with gentle stirring. The array was washed 3 times with TBS-T. The array was incubated at room temperature for 1 hour with gentle stirring with 70mL of a solution containing streptavidin-Cy5 conjugate. The array was washed 3 times with TBS-T, rinsed quickly in deionized water, and dried.

[0224] The slides can be imaged in the PERKIN-ELMER SCANARRAY 4000 confocal fluorescence imaging system. The array images can be saved and analyzed using the imaging research ARRAYVISION software. In short, the spot intensity is determined by subtracting the background signal. The spot replicates from each sample condition can be averaged and then compared with the appropriate control.

[0225] Endothelial Tube Formation: Endothelial tube formation is associated with angiogenesis and the formation of microvascular capillaries. Capillary formation and angiogenesis may contribute to skin redness and rosacea. The ability of endothelial cells to form tubes can be determined using a capillary tubule disruption assay with preformed primary human umbilical vein endothelial cells (HUVEC) in a cell culture system in the presence or absence of test extracts and compounds.

[0226] Briefly, HUVECs are cultured in vitro on an extracellular matrix, which stimulates the attachment and tubular morphogenesis of endothelial cells, forming capillary-like luminal structures. These in vitro formed capillaries are similar to human vascular capillaries in many aspects. The capillary assay is based on this phenomenon and is used to evaluate potential vascular targeting agents.

[0227] HUVEC cultures were grown in a 5% CO2 37°C cell culture incubator. The complete growth medium for HUVEC was endothelial basal medium (EBM) supplemented with 2% fetal bovine serum (FBS), 12 μg / mL bovine brain extract, 1 μg / mL hydrocortisone, and 1 μg / mL GA-1000 (gentamicin-amphotericin). HUVEC cultures between passages 3 and 8 were used for all analytical experiments.

[0228] HUVECs are pre-labeled with the fluorescent agent Calcein AM and seeded in 96-well culture plates coated with extracellular matrix containing their complete growth medium. After approximately four hours of the morphogenesis process, endothelial capillaries should be formed. Then, a 50 μL volume of the designed dose of the test agent is applied as a treatment condition to the formed capillary culture. Untreated controls can be added with the vehicle of the test agent. SUTENT ® is an FDA-approved anti-angiogenic drug, and its concentration can be used as a control for assay performance. After approximately six hours of treatment, the morphology of endothelial tubules in each well is examined by microscopy, imaging, and the capillary disrupting activity under the treatment conditions can be quantified. Each test condition can be performed in duplicate wells, including controls.

[0229] * * * * * * * * * * * * * *

[0230] All compositions and / or methods disclosed and claimed herein can be made and implemented according to the present disclosure without undue experimentation. Although the compositions and methods of the present invention have been described in accordance with preferred embodiments, it is obvious to those skilled in the art that changes can be implemented in the compositions and / or methods and in the steps or the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the present invention. More specifically, it is obvious that specific reagents related to both chemical and physiological aspects can replace the reagents described herein while achieving the same or similar results. All such similar substitutions and changes that are obvious to those skilled in the art are considered to be within the spirit, scope and concept of the present invention as defined by the appended claims.

Claims

1. A method for alleviating skin inflammation in a subject, comprising topically applying a cosmetic preparation, wherein the cosmetic preparation comprises at least one transient receptor potential channel vanilloid subtype 1 (TRPV-1) antagonist, wherein the at least one TRPV1 antagonist is a saccharide isomer, a reed extract and a Poria cocos extract, and / or a cucurbita pekinensis (pumpkin) seed extract.

2. The method according to claim 1, wherein the cosmetic formulation comprises more than one TRPV-1 antagonist.

3. The method of claim 1, wherein the cosmetic formulation comprises at least 1 weight / volume % of saccharide isomers, at least 5 weight / volume % of cucurbita pepo (pumpkin) seed extract, and / or at least 1 weight / volume % of reed extract and Poria cocos extract.

4. The method of claim 1, wherein the topical administration results in inhibition of cellular calcium flux when compared to an appropriate relative control.

5. The method of claim 1, wherein the skin inflammation and / or irritation is or was caused by agonist activation of TRPV-1.

6. The method according to claim 5, wherein the agonist of TRPV-1 is hydroquinone, resorcinol, trichloroacetic acid, a carboxylic acid, a dicarboxylic acid, a β-hydroxy acid and / or an α-hydroxy acid.

7. The method of claim 8, wherein the α-hydroxy acid is citric acid, glycolic acid, lactic acid, malic acid, mandelic acid and / or tartaric acid; and / or wherein the β-hydroxy acid is salicylic acid, a derivative or metabolite of salicylic acid, β-hydroxybutyric acid, tropic acid and / or tresocarboxylic acid.

8. The method according to claim 7, wherein the agonist of TRPV-1 is retinol, a retinol precursor, a retinol derivative and / or a retinol metabolite.

9. The method of claim 14, wherein the formulation comprising retinol (vitamin A), a retinol precursor, a retinol derivative, or a retinol metabolite comprises at least about 0.1% w / v to about 1% w / v of the ingredient.

10. The method according to claim 8, wherein the formulation comprising retinol and the cosmetic formulation comprising at least one TRPV-1 antagonist are the same composition.

11. The method of claim 1, further comprising administering a formulation comprising at least one inhibitor of a pro-inflammatory cytokine.

12. The method of claim 11, wherein the at least one pro-inflammatory cytokine inhibitor comprises rosemary leaf extract.

13. The method according to claim 11, wherein the formulation comprising at least one proinflammatory cytokine inhibitor and the cosmetic formulation comprising at least one TRPV-1 antagonist are the same composition and, optionally, the composition further comprises retinol.

14. The method of claim 13, wherein the expression of interleukin-6 (IL-6), interleukin-8 (IL-8), and / or tumor necrosis factor alpha (TNFα) is inhibited when compared to an appropriate relative control.

15. The method of claim 1, wherein topical application of the cosmetic formulation reduces erythema, edema, scaling, dryness, burning, stinging, and / or itching.

16. The method of claim 1, wherein topical application of the cosmetic formulation results in improved skin pigmentation, reduced fine lines and wrinkles, improved evenness of skin tone, reduced photodamage, and / or improved skin firmness when compared to an appropriate relative control.

17. The method of claim 1, wherein the cosmetic formulation is formulated for administration at least every other day for at least 4 weeks and / or at least 8 weeks.

18. The method according to claim 17, wherein the cosmetic preparation is formulated as a leave-on product or a rinse-off product.

19. A cosmetic preparation comprising At least one TRPV1 antagonist selected from reed extract and Poria cocos extract, saccharide isomers and zucchini (pumpkin) seed extract, at least one cytokine inhibitor, and At least about 0.1 wt / vol % to about 1 wt / vol % or about 0.4 wt / vol % to about 0.6 wt / vol % retinol.

20. The cosmetic formulation according to claim 19, wherein the composition comprises at least 1 weight / volume % of saccharide isomers, at least 5 weight / volume % of Cucurbita pekinensis (pumpkin) seed extract and / or at least 1 weight / volume % of Phragmites australis extract and Poria cocos extract.

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