TW201705937A - Compositions including obacunone and methods of using the same in skin anti-aging applications - Google Patents

Abstract

Compositions for topical use and having a protein carbonylation-inhibiting activity that include an amount of about 75-100% pure obacunone effective to reduce and/or inhibit protein carbonylation are described. Methods for inhibiting protein carbonylation in skin and for skin whitening and lightening are also described.

Description

Composition comprising corkone and method of using the same in skin anti-aging applications Field of invention

Described herein are compositions comprising baconone and their use in methods of inhibiting skin protein carbonylation, as well as methods of improving skin appearance or improving at least one sign of skin aging, such as yellowing of the skin.

Background of the invention

Oxidative stress is associated with non-enzymatic production of reactive oxygen species in a reaction known as protein carbonylation. Protein carbonylation is a term used to describe a process in which a reactive aldehyde or ketone is introduced into a protein by oxidation. Protein carbonyl has been shown to be a major product of protein oxidation and can be formed by oxidative cleavage of proteins, direct oxidation of amino acid residues or covalent reaction with aldehydes derived from lipid peroxidation.

Protein carbonylation is due to direct metal catalyzed oxidation of the amino acid side chain (primary protein carbonylation) or reactive aldehyde and amino acid side chain addition (secondary protein carbonylation). Thus, it is known that exogenous reactive aldehydes such as acrolein are derived from cigarette smoke. However, reactive aldehydes can also be formed endogenously, for example, polyunsaturated everywhere in the human body. Oxidative degradation of fatty acids can result in the formation of various aldehydes such as acrolein and 4-hydroxynonenal (4-HNE).

It has previously been reported that primary protein carbonylation plays an important role in the mechanism of reactive oxygen species (ROS) signaling. Specifically, primary protein carbonylation mediates cellular signaling and primary protein carbonylation is reversible.

Primary protein carbonylation involves direct metal catalyzed oxidation of the amino acid side chains, especially on the following four sensitive amino acid residues: valine, arginine, lysine and threonine. Oxidation of the proline or arginine side chain results in the formation of a glutamic acid semialdehyde, which oxidizes to produce an aminoadipate semialdehyde, thereby introducing a carbonyl group into the protein structure. Oxidation also converts the hydroxyl group of the threonine side chain to a carbonyl group to form 2-amino-3-ketobutyric acid. Secondary protein carbonylation mechanisms include Michael addition of reactive aldehydes to the side chains of lysine, histidine and cysteine residues, and reduction of sugars to lysine and amino groups on arginine Alkali, leading to an increase in the progressive glycation end product. Carbonyl proteins are present in a variety of oxidative stresses and conditions. In addition to inactivated proteins, carbonylated cellular proteins undergo proteasome-dependent degradation. Primary protein carbonylation is believed to play an important role in labeling damaged proteins to remove them from biological systems during oxidative stress.

It has been well documented that the production of carbonyl proteins is a hallmark of oxidative stress, for example due to exogenous factors such as exposure to ultraviolet radiation or topical oxidative chemicals, or due to endogenous The factor is the chemical attack by the reactive carbonyl derived from the degradation of the lipid peroxide.

Photodamaged facial skin has a variety of unique features, such as black Spots, wrinkles and sagging. Older people, especially Asians, tend to turn yellow with light aging skin. Among Chinese, yellowing of the face means dim or not matte. Common clinical signs of photodamage or aging include the appearance of a sallow or yellow appearance of the skin. It has been shown that carbonylation of the skin occurs both in the epidermal layer of the skin as well as in the dermis layer. In addition, carbonylation of the skin often results in significant photodamage of skin tissue and structure that is subject to oxidative stress, which, if not prolongedly prevented or treated, can lead to irreversible discoloration: yellowing of the face.

Accordingly, there is a great need for anti-aging compositions and methods that inhibit or prevent protein carbonylation to improve skin appearance or to improve at least one sign of skin aging (eg, yellowing of the skin) and to enhance skin brightness of facial skin.

Summary of invention

One embodiment is directed to a topical composition having reduced or inhibited protein carbonylation activity consisting essentially of about 75-100% pure phellodone in an amount effective to reduce or inhibit protein carbonylation; a cosmetically acceptable vehicle And optionally, one or more cosmetic ingredients selected from the group consisting of alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosity modifiers, emulsifiers, stabilizers, colorants, and a fragrance; and/or a skin lightening agent selected from the group consisting of tyrosinase inhibitors, free radical scavengers, and mixtures thereof.

Another embodiment is directed to a topical composition having reduced or inhibited protein carbonylation activity consisting of about 90% pure phellodone in an amount effective to reduce or inhibit protein carbonylation; a cosmetically acceptable vehicle; Optionally, one or more cosmetic ingredients selected from the group consisting of alcohols, fats, Oil, surfactant, fatty acid, emu oil, wetting agent, moisturizer, viscosity modifier, emulsifier, stabilizer, colorant and fragrance; and/or skin lightening agent selected from the group consisting of: tyrosinase inhibitors, free Base scavengers and mixtures thereof.

In the composition, the corkone is purified from an extract of at least one of the following plants selected from the group consisting of Evodia rutaecarpa, Dictamni cortex, and Phellodendri amurensis. Cortex), bergamot (Citri sarcodactylis fructus), argus (Aurantii frusctus immaturus) and citron (Citri fructus). The extract of Evodia rutaecarpa is normalized to the content of at least one of evodiamine, rutaecarpine and limonin. In the composition, the ketone is about 99.99% pure phellodone active. The composition is a preparation selected from the group consisting of creams, ointments, foams, lotions, plasters, tablets, granules or emulsions. In the composition, the amount of protein carbonylation effective to reduce or inhibit is from about 10 to 1000 mg. In the composition, the cedarone comprises from about 1 to 99% w/w of the composition. Alternatively, the cedarone comprises from about 1% to about 75% w/w of the composition. Alternatively, the cedarone comprises about 50-70% w/w of the composition.

Another embodiment is directed to a topical composition comprising a cosmetically acceptable vehicle and an amount of at least 90% pure cedarone, wherein the cedarone comprises from about 1 to 99% w/w of the composition.

Other embodiments are directed to a method of improving the appearance of the skin or improving at least one sign of skin aging, the method comprising topically applying the composition to the skin in a cosmetically effective amount sufficient to improve the appearance of the skin or at least An indication of skin aging, wherein the composition comprises an amount of about 75-100% pure phbodone, a cosmetically acceptable vehicle, and, optionally, one or more cosmetic ingredients selected from the group consisting of: alcohols, fats , oils, surfactants, fatty acids, eucalyptus oils, wetting agents, humectants, viscosity modifiers, emulsifiers, stabilizers, colorants and perfumes; and/or skin lightening agents selected from the group consisting of: tyrosinase inhibitors, Free radical scavengers and mixtures thereof. In the method, the composition is in the form of a product selected from the group consisting of aerosols, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, moisturizing Hair oil, solution, pump spray, sticks, small towels and combinations thereof.

Another embodiment is directed to a method of whitening skin comprising topically applying a composition described herein to the skin, wherein the composition is applied in an amount and for a duration sufficient to visibly whiten the skin.

Another embodiment is also directed to a method of brightening skin comprising topically applying a composition described herein to the skin, wherein the composition is applied in an amount and for a duration sufficient to visibly brighten the skin.

Other embodiments are directed to the use of a topical composition as described herein in the manufacture of a cosmetic agent having reduced or inhibited protein carbonylation activity.

Another embodiment is directed to a method of preventing, ameliorating or reducing at least one sign of skin aging comprising applying a composition described herein to the skin, wherein the composition is applied in an amount and for a duration sufficient to prevent, ameliorate or reduce at least one Signs of skin aging.

Figure 1 depicts a graph showing corkone in HepG2 cells Low cytotoxicity - CTG.

Figure 2 depicts a graph showing the dose-dependent inhibition of protein carbonylation by phellodone in HepG2 cells.

Detailed description of the invention

It is to be understood that the invention is not limited to the particular compositions, methods or aspects described herein. Moreover, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art It is also understood that the terminology used herein is for the purpose of describing the particular embodiments and is not intended to limit the scope of the invention. The present invention is based on the surprising discovery that corkone has a positive effect as a skin lightening agent and is non-toxic (Fig. 1) and has an inhibitory effect on protein carbonylation (Fig. 2). As shown in Figure 2, protein carbonylation was inhibited with increasing dose of ketone.

Accordingly, described herein are compositions and their use in anti-aging, whitening or brightening methods for the skin comprising purified piloinone in an amount sufficient to effect reduction or inhibition of protein carbonylation.

definition

All percentages and ratios are by weight unless otherwise indicated.

The term "composition" refers to a product that treats, ameliorates, promotes, enhances, manages, controls, maintains, optimizes, modifies, reduces, inhibits, or prevents a particular state associated with a natural state, biological process, or disease or disorder. The term "composition" includes, but is not limited to, pharmaceutical (ie, drugs), over-the-counter drugs. An (OTC), cosmetic, food, food ingredient or dietary supplement composition comprising an effective amount of a purified compound, an extract, a natural complex, at least one component thereof, or a mixture thereof. Exemplary compositions include compositions in the form of creams, cosmetic waters, packs or powders or emulsions, lotions, elixirs, foams, tablets, plasters, granules or ointments.

The term "extract" as used herein, refers to a solid, semi-fluid or liquid substance or formulation comprising an active ingredient component of a plant, such as scutellaria in concentrated form. The term "extract" is intended to include not only a crude extract prepared from a plant using a solvent, but also a fraction of the crude extract in the solvent selected from the group consisting of water, 1 to 4 carbon atoms. Lower alcohol (eg methanol, ethanol, butanol, etc.), ethylene, acetone, hexane, diethyl ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N,N-dimethylformamide (DMF) , dimethyl hydrazine (DMSO), 1,3-butanediol, propylene glycol, and combinations thereof. Any extraction method can be employed as long as the extraction and preservation of the active ingredient can be ensured.

The term "purified" as used herein in connection with cedarone refers to the preparation of cedarone by physically separating the cedarone from the remaining components of the plant extract. Various purification methods are known and can be used to obtain pure phbodone. For example, methods such as fractional distillation or affinity purification can be used. As a result of the purification process, the ketone can be at least about 10% pure; or, at least about 20% pure; or, at least about 30% pure; or, at least about 40% pure; or, at least about 50% pure; or, At least about 60% pure; or, at least about 70% pure; or, at least about 80% pure; or, at least about 90% pure; or, at least about 95% pure; or, at least about 99% pure; but preferably about 75 %-100% pure.

The term composition, extract, extract mixture, An "effective amount" or "therapeutically effective amount" of an extract component and/or an active agent or active ingredient refers to an amount that is effective at a dose and that lasts long enough to achieve the desired result. For example, the "effective amount" or "therapeutically effective amount" means when the composition, extract, extract mixture, extract component and/or active agent or active ingredient of the present invention is administered to a subject ( For example, a mammal, such as a human, is in an amount sufficient to produce a therapeutic effect, such as a decrease or inhibition, in the subject, the extract, the extract mixture, the extract component, and/or the active agent or active ingredient. Protein carbonylation, skin anti-aging, improve or improve skin whitening and / or brighten, reduce or dilute dark spots and so on. The amount of the disclosed compositions, extracts, extract mixtures, extract components and/or active agents or active ingredients that constitute "effective amount" or "therapeutically effective treatment" will be treated according to the active agent or compound, The condition and its severity, the mode of administration, the duration of treatment, or the age of the subject to be treated will vary, but it can be routinely determined by one of ordinary skill in the art in light of its own knowledge and the content of the present disclosure.

The terms "preventing, ameliorating or reducing at least one sign of aging" and "preventing, ameliorating or reducing signs of aging" are used interchangeably herein to indicate preventing, preventing, reversing, alleviating, reducing, and/or reducing signs of aging. Representative signs of aging include, but are not limited to, skin yellowing, lines, fine lines, wrinkles, crow's feet, dark circles, moles, age spots, stretch marks, or combinations thereof.

"Improving the appearance of the skin" and "Improving the aesthetics of the skin" are used interchangeably herein to refer to an aesthetic improvement in the appearance of the skin. Representative improvements may include, but are not limited to, skin tone, tone, texture, shine, Favorable properties related to brightness, gloss, whiteness, cleanliness, thickness, elasticity, toughness, moisture retention, smoothness, contour, hardness, firmness, flexibility, softness, sensitivity, pore size or combinations thereof / or nature. These terms can also be used to refer to an improvement in adverse skin conditions. Representative adverse conditions resulting from the adverse skin condition, resulting in or resulting from the unfavorable skin condition include, but are not limited to, skin aging, psoriasis, eczema, seborrhea, dermatitis , sunburn, estrogen imbalance, hyperpigmentation, hypopigmentation, discoloration, yellowing, freckles, skin atrophy, skin rash, skin fragility, dryness, rough touch, cracking, sagging, thinning, hyperplasia, fibrosis , enlarged pores, cellulite formation, bruises, acne formation, apoptosis, cell differentiation, cell dedifferentiation, prevention of tumor induction or tumor progression, viral infection, fungal infection, bacterial infection, arachnoid vascular disease (capillaries) Telangectasia, hirsutism, rosacea, pruritus, warts, warts, corns, or combinations thereof.

The term "pharmaceutically acceptable" means those drugs, pharmaceuticals, extracts or inert ingredients which are suitable for use in combination with humans and lower animals without undue toxicity, incompatibility, instability, irritation, and the like. Proportional to a reasonable benefit/risk ratio.

The terms "administering" and "administering" are defined to provide a composition to a subject by routes known in the art including, but not limited to, intravenous, intraarterial, oral, parenteral, buccal, topical. , transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal or intraperitoneal routes of administration. In a preferred embodiment, it is suitable to administer the composition in a topical manner.

The term "subject" or "individual" as used herein includes a mammal to which the composition can be administered. Non-limiting examples of mammals include humans, non-human primates, rodents (including transgenic and non-transgenic mice), and the like. In some embodiments, the subject is a mammal, and in some embodiments, the subject is a human.

Corkone

Phellodone (CAS: 751-03-1) is a compound having a molecular weight of 454.52 (i.e., a botanical ingredient) which can be extracted from Evodia rutaecarpa BENTHAM (Ruphaceae). For example, the fruit of Rutaceae can be extracted with EtOH to obtain an extract of Evodia rugulosa ("Wuhan extract") which can be used as a source of a phbodone compound which is used in the unique composition of the present invention. The term "corkone" refers to plant constituents as well as plant extracts or natural complexes.

An exemplary structure of corkone (C ₂₆ H ₃₀ O ₇ ) is as follows:

Phellodone is commercially available from a variety of sources including ChemStrong Scientific Co., Ltd or Chengdu Biopurify Phytochemicals Ltd. Further, the cedarone can be purified from an extract of a plant including, for example, dried root bark of Evodia rugulosa, Rhizoma (Ruphal), dried skin of Phellodendron amurense Rupr. (Fam. Rutaceae), Chuanhuang Phellodendronchinense Schneid. (Fam. Rutaceae) dry skin, Citrus sp. Fortunella margarita, Casimiroaedulis, bergamot, medlar and citron. In some aspects, the flowers, flower heads or fruits of the plants, or leaves are used as a source of extract. When extracted, the extract can be standardized as an active labeling compound. For example, the Cortex extract can be normalized to the concentration of at least one of berberine, bamartine, limonin, and cedarone.

Various purification and extraction techniques are discussed more fully below or are known in the art.

Exemplary extraction process

In one embodiment, the extracts and/or active compounds used in the unique compositions of the invention can be obtained using organic solvent extraction techniques.

In another embodiment, continuous fractionation using a solvent such as H ₂ O, H ₂ O-EtOH, H ₂ O-MeOH or H ₂ O-acetone as a solvent can be used to obtain an extract for use in the unique composition of the present invention. And / or active compound.

The total hydro-ethanolic extraction technique can also be used to obtain extracts and/or active compounds for use in the unique compositions of the present invention. Usually this is called a one-time extraction. The extract produced in this process will contain a wide variety of phytochemicals, including fat soluble and water soluble materials, present in the extracted material. After collecting the extract solution, the solvent was evaporated to obtain the extract.

Total ethanol extraction can also be used in the present invention. Used by this technology It is ethanol, not water-ethanol as a solvent. The extract obtained by this extraction technique may contain, in addition to the water-soluble compound, a fat-soluble and/or lipophilic compound.

Another example of an extraction technique that can be used to obtain extracts and/or active compounds for use in the present invention is supercritical fluid carbon dioxide extraction (SFE). In this extraction step, the substance to be extracted is not exposed to any organic solvent. Instead, the extraction solvent is carbon dioxide with or without a regulator and under supercritical conditions (>31.3 ° C and >73.8 bar). Those skilled in the art will appreciate that temperature and pressure conditions can be varied to obtain an extract of optimal yield. This technique results in an extract of a fat-soluble and/or lipophilic compound similar to the above-described perhexane and ethyl acetate extraction techniques.

Those skilled in the art will appreciate that there are many other extraction and purification processes that can be used to obtain the extracts and/or active compounds used in the practice of the present invention, which are known in the art and in various patents and publications. The process described in the article. For example, the extraction operations described in the following references can be used in the practice of the present invention, which is incorporated herein by reference: Murga et al., "Extraction of natural complex phenols and tannins from grape seeds by using supercritical mixtures of carbon dioxide And alcohol.” J. Agric Food Chem. 2000 August: 48(8): 3408-12; Hong et al., “Microwave-assisted extraction of phenolic compounds from grape seed.” Nat Prod Lett. 2001; 15(3): 197-204; Ashraf-Khorassani et al., "Sequential fractionation of grape seeds into oils, polyphenols, and procyanidins via a single system employing CO ₂ -based fluids." J. Agric Food Chem., 2004 May 5; 52(9) :2440-4.

According to one embodiment of the invention, the corkketone extract can be prepared by extracting the Eel with the 70% EtOH. The final extract had a water content of >5.0% to produce a plant extract having a natural extraction rate of plants: finished product = 5-7:1.

The corkone extract is then assayed and normalized to the content of active markers such as evodiamine, rutaecarpine, berberine, babatin and limonin.

The corkone active is then purified by solvent-water partitioning, such as a dichloromethane-water, ethyl acetate-water, n-butanol-water system.

Alternatively, the corkone active can be purified by various chromatographic methods, such as macroporous resins, fluorenone resins, MCI resins, gels, and the like.

Further, the corkone active may be purified by preparative chromatography, semi-preparative chromatography (for example, preparative HPLC, semi-preparative HPLC) or the like.

Additionally, the corkone active can be purified by high speed reflux chromatography.

combination

The cedarone can be formulated into the composition. In particular, some embodiments are directed to compositions having reduced or inhibited protein carbonylation activity comprising an amount of cedarone effective to reduce or inhibit protein carbonylation. Some other embodiments relate to compositions for skin whitening and/or skin lightening comprising an amount of corkone that effectively whitens or brightens the skin.

Some other embodiments relate to compositions that are resistant to skin aging, Contains an amount of cedarone effective to prevent or treat skin aging.

Some embodiments are directed to a composition that reduces or inhibits protein carbonylation, comprising an amount of cedarone effective to reduce or inhibit protein carbonylation, wherein the cedarone is at least about 75% pure. Alternatively, in the composition, the ketone is at least about 80% pure; or, at least about 85% pure; or, at least about 90% pure; or, at least about 95% pure; or, at least about 96% pure; Alternatively, at least about 97% pure; or, at least about 98% pure; or at least about 99% pure; or, in the composition, the ketone is about 99.99% pure.

The cedarone comprises about at least 50% w/w of the composition, more preferably about at least 55% w/w of the composition, about at least 60% of the composition, and about at least 65 of the composition. % w/w, about at least 70% w/w of the composition, about at least 75% w/w of the composition, about at least 80% w/w of the composition, and about the composition. At least 85% w/w, about at least 90% w/w of the composition, about at least 95% w/w of the composition, and about at least 99% w/w of the composition. Alternatively, the cedarone comprises from about 1% to about 99% w/w of the composition; alternatively, the cedarone comprises from about 1% to about 90% w/w of the composition; or, the cedarone comprises from about 1% to about 75% of the composition w/w; alternatively, the cedarone comprises about 50-70% w/w of the composition.

In the composition, the ketone can be present in an amount from about 0.001% to about 10%.

Of course, those skilled in the art will appreciate that the amount of corkone incorporated into the composition can depend on the amount of standardized marker. For example, for the cedarone prepared from the extract of Evodia rutae, the marker may be evodiamine, rutaecarpine and/or limonin. Therefore, if the ketone is labeled To normalize to about 40% of the labeled compound, the amount of the above-described cedarone will be halved. For example, the corkone extract can be standardized to about 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60% , 65%, 70%, 75%, 80%, 85%, 90% or 95% labeled compound content.

The composition may have a pH of from about 6.0 to about 8.0, or the composition may have a substantially neutral pH. In some embodiments, the composition in which the cedarone is used has a substantially neutral pH.

Some embodiments relate to compositions for enhancing skin whitening or lightening. The composition contains an effective amount of cedarone to enhance skin whitening or lightening. The composition can comprise a cosmetically acceptable vehicle.

Some other embodiments are directed to methods of enhancing skin whitening or brightening comprising administering to a subject a composition comprising a cedarone extract, wherein the composition reduces or inhibits protein carbonylation of the skin, thereby enhancing skin whitening or Brighten up.

The compositions of the present invention may be formulated in acceptable carriers, vehicles or excipients and may be prepared, packaged and labeled as a cosmetic or pharmaceutical product for use in the skin whitening or lightening of the subject. In some embodiments, the cedarone comprises about 50-70% w/w of the composition, and the carrier, vehicle or excipient comprises about 50-30% w/w of the composition.

Typically, the composition will comprise a cosmetically acceptable vehicle. Examples of cosmetically acceptable vehicles suitable for use in all embodiments of the invention include, but are not limited to, water; glycerin; various alcohols (e.g., ethanol, propanol); Vegetable oil; mineral oil; eucalyptus oil; fatty ether; fatty acid ester; fatty alcohol; diol; polyglycol or any combination thereof. Methods of formulation are well known in the art and are disclosed, for example, in Remington's Pharmaceutical Sciences, Gennaro, Mack Publishing Co., Easton Pa., 1990, which is incorporated herein by reference.

In some embodiments, the composition may further comprise one or more whitening agents to enhance the whitening effect of the composition. Exemplary whitening agents include all known whitening agents and those that can be developed in the future. Preferably, the whitening agent is selected from the group consisting of: a tyrosinase inhibitor, a free radical scavenger, and mixtures thereof. Examples of suitable tyrosinase inhibitors include, but are not limited to, kojic acid and its derivatives, arbutin and its derivatives, licorice extract and its derivatives, ascorbic acid and its derivatives, and hydroquinone and its derivatives. Things. Examples of suitable free radical scavengers include, but are not limited to, licorice extract and its derivatives, vitamin E and its derivatives, vitamin A and its derivatives, vitamin C and its derivatives, rosemary extract and Derivatives and superoxide dismutase. Preferred whitening agents are selected from the group consisting of: kojic acid, kojic acid derivatives, arbutin, arbutin derivatives, bearberry extract, lemon extract, cucumber extract, vitamin C and its derivatives, and acerola cherry extract. And acerola cherry ferment.

Non-exclusive examples of vitamin C derivatives are, for example, alkyl esters of L-ascorbic acid, such as L-ascorbyl palmitate, L-ascorbyl isopalmitate, L-ascorbyl dipalmitate, L-ascorbyl isostearic acid Ester, L-ascorbyl distearate, L-ascorbyl diisostearate, L-ascorbyl myristate, L-ascorbyl isomyristate, L-ascorbic acid 2-ethylhexanoate, L -ascorbic acid-di-2-ethylhexanoate, L-antibody Ascorbyl oleate and L-ascorbic acid dioleate; phosphates of L-ascorbic acid, such as L-ascorbyl-2-phosphate and L-ascorbyl-3-phosphate; sulfate of L-ascorbic acid, For example, L-ascorbyl-2-sulfate and L-ascorbyl-3-sulfate; salts formed with alkaline earth metals such as calcium and magnesium. They may be used singly or in the form of a mixture of two or more.

When the corkone is mixed with any known whitening agent, the ratio of the corkone to the known whitening agent is from about 1:100 to about 100:1, preferably from about 1:50 to about 50:1, more Choose from about 1:10 to about 10:1. Most preferably, the ratio of corkone to known whitening agents is from about 1:5 to about 5:1.

The compositions of the present invention can be made into a variety of forms. For example, they may be in the form of a cosmetic preparation, such as in the form of a cream, cosmetic water, facial mask or powder, or as an emulsion, lotion, tincture, foam, tablet, plaster, granule, aerosol spray, coagulation In the form of a gel, mousse, balm or ointment. In each of the formulations, various known conventional cosmetic ingredients can be combined. For example, cosmetic ingredients such as alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosity modifiers, emulsifiers, stabilizers, colorants, and perfumes may be included. Some other embodiments are directed to compositions which can be formulated into any convenient form suitable for topical application to the skin. Suitable emulsions include oil-in-water, water-in-oil, and water-in-silicone emulsions.

In some embodiments, an improved type of whitening skin composition comprising a skin lightening agent is provided, wherein the improvement is the addition of corkone. The composition can comprise a cosmetically acceptable vehicle.

The compositions of the invention may be administered topically.

The composition can be administered once a day, administered as many times as needed or Administration is carried out in any suitable dosage regimen to achieve the desired effect. The composition can be administered continuously or intermittently.

method

The compositions described herein can be used in a variety of cosmetic and/or pharmaceutical applications, including inhibiting or reducing protein carbonylation and/or enhancing or improving skin whitening, brightening, and whitening.

Some embodiments relate to a method of whitening skin comprising topically applying to any combination of any of the above compositions, and any combination thereof.

Some embodiments relate to a method of whitening skin comprising topically applying a composition to the skin, the composition comprising an amount of cedarone effective to reduce or inhibit protein carbonylation in the skin. The amount and duration of application of the composition is sufficient to visually whiten the skin.

In a topical whitening skin cosmetic composition containing one or more cosmetic ingredients selected from the group consisting of alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosity modifiers, emulsifiers , Stabilizers, Colorants, and Perfumes, the improvement consisting in including an effective amount of cedarone to whiten the skin.

In this method, the frequency of topical application will depend on a variety of factors, including the desired degree of inhibition of protein carbonylation and the amount of cedarone in the composition. One suitable exemplary dosing regimen includes administration to the skin twice daily, once in the morning and evening.

Other embodiments are directed to a method of improving the appearance of skin or at least one sign of skin aging, the method comprising topically applying to the skin a cosmetically effective amount of an amount sufficient to improve the appearance of the skin or at least one sign of skin aging Composition, wherein the composition comprises an amount of about 75-100% pure phbodone; a cosmetically acceptable vehicle; and, optionally, one or more cosmetic ingredients selected from the group consisting of alcohols, fats, Oil, surfactant, fatty acid, emu oil, wetting agent, moisturizer, viscosity modifier, emulsifier, stabilizer, colorant and fragrance; and/or skin lightening agent selected from the group consisting of: tyrosinase inhibitors, free Base scavengers and mixtures thereof. In the method, the composition is in the form of a product selected from the group consisting of aerosols, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, moisturizing Hair oil, solution, pump spray, sticks, small towels and combinations thereof.

Some embodiments relate to the use of a topical composition as described herein for the preparation of a cosmetic agent having reduced or inhibited protein carbonylation activity.

Some other embodiments are directed to a method of preventing, ameliorating or reducing at least one sign of skin aging comprising applying any of the above compositions to the skin, wherein the composition is applied in an amount and for a duration sufficient to prevent, ameliorate or reduce the at least one skin Signs of aging. Representative signs of aging include, but are not limited to, skin yellowing, pleats, fine lines, wrinkles, crow's feet, dark circles, moles, age spots, or combinations thereof.

Example

Example 1: Cytotoxicity/safety evaluation of corkone by CellTiter-Glo® (CTG) luminescent cell viability test in HepG2 cells

In order to determine the cell safety of phellodone, the following tests were conducted.

CellTiter-Glo® (CTG) according to the manufacturer's instructions The luminescent cell viability test (obtained from Promega) was used to determine the number of viable cells in the medium based on the quantification of ATP (indicator of metabolically active cells) present. 99.99% pure cedarone and hydroquinone with the same gradient concentration (100, 33.3, 11.1, 3.7, 1.23, 0.41, 0.14, 0.05, 0.02 (ug/mL)) were used in this test (used as a reference) . This test was performed using HepG2 cells.

Phellodone showed no cytotoxicity by cell-based safety assessment by CTG (see Table 1).

The result is shown in Figure 1.

Example 2: Phellodone has been shown to reduce protein carbonylation in a dose-dependent manner in HepG2 cells

To determine whether obacunone Protein carbonylation, the following cell-based assay, the test using OxiSelect ^TM Protein Carbonyl ELISA kit (Cell Biology # STA-310) .

HepG2 cells (ATCC) were used for this test. The cells were seeded in plates at 100,000 cells per well and the test plates (96-well plates obtained from Costa) were incubated for 24 hours at 37 ° C, 5% CO ₂ in humid conditions.

0 ug/mL, 33 ug/mL, 100 ug/mL, and 300 ug/mL of cedarone were tested. Vitamin C (a well-known protein carbonylation inhibitor) of 0 ug/mL, 33 ug/mL, 100 ug/mL, and 300 ug/mL was used as a positive inhibitor control in this test.

A reference and test compound solution (200x) was prepared according to a platemap. The phbodone was dissolved in MEM (lowest basal medium) and added to HepG2 cells seeded 24 hours before treatment as described above. The final concentration of this test compound was 1x (Table 2). The plate was then incubated for 72 hours at 37 ° C, 5% CO ₂ in humid conditions.

On day 3, the medium was removed from the cells. The wells were washed twice with HBSS (Hanke's Balanced Salt Solution). 100 μM 8-OH quinoline and 100 μM FeCl _{3 were} added to the wells and the plate was incubated for 1 hour. The cells were then washed once with HBSS. After the washing step, fresh medium is added to each well. The plate was then incubated for 72 hours at 37 ° C, 5% CO ₂ in humid conditions.

On day 4, the medium was removed from the cells and the cells were washed twice with HBSS. 100 μL HBSS (with mixture (commercially available) A protein acid inhibitor mixture tablet (Roche #04 693 124 001)) was added to each well. The plate was then placed at -80 °C for 30 minutes, thawed and the freeze/thaw step repeated at least 3 times. The cell lysate was then mixed and tested for 25 μL of cell lysate to determine the protein concentration. Table 3 shows the preparation of the BCA kit to determine the protein concentration on day 4.

Then adjust the protein concentration. 100 [mu]L of sample (using the induced BSA dilution) was then added to the 96-well protein binding plate and incubated overnight at 4[deg.]C.

On day 5, an ELISA test was performed. The wells were washed 3 times with 250 μL of 1X PBS (phosphate buffered saline) per well. Excess wash solution is then removed from each well. 100 μL of DNPH (dinitrophenylguanidine) working solution was added to each well (Table 4) and the plate was incubated at room temperature for 45 minutes in the dark.

The wells were then washed 5 times with 250 [mu]L IX PBS/ethanol (1:1) and incubated for 5 minutes on an orbital shaker (Table 5).

The well was then washed twice with 250 μL of 1X PBS. 200 μL of blocking solution was added to each well (Table 6) and the plate was incubated for 1-2 hours at room temperature on an orbital shaker.

The wells were then washed 3 times with 250 [mu]L of 1X wash buffer (Table 7) with thorough aspiration between each wash.

Next, 100 μL of anti-DNP (anti-dinitrophenylguanidine) antibody was added to each well (Table 8) and the plate was incubated for 1 hour at room temperature on an orbital shaker.

Next, the strip holes were washed 3 times (1* wash buffer). 100 μL of diluted HRP bound to secondary antibody was added to all wells and the plate was incubated for 1 hour at room temperature on an orbital shaker. See Table 9.

Next, the strip holes were washed 5 times (1* wash buffer). The substrate solution was warmed to room temperature and 100 μL of this substrate solution was added to each well. The strip wells were then incubated for 15 minutes. The enzyme reaction was terminated by adding 100 μL of stop solution to each well. Once the color fades over time, the result is read immediately.

The results are shown in Table 10 and Figure 2.

The average protein carbonylation inhibition rate of cedarone was 1.6%, 23.6%, 37.75%, and 39.15%; the inhibition of carbonylation of vitamin C was -3.05%, 31.05%, 55.35%, and 62.65%.

This indicates that in HepG2 cells, corkone has excellent potency against protein carbonylation compared to the known positive control vitamin C. It also shows that products containing corkone can be used to brighten the skin and resist aging.

Therefore, the foregoing detailed description is to be considered as illustrative and not restrict

Claims (18)

A topical composition having activity to reduce or inhibit protein carbonylation, consisting essentially of: a) about 75-100% pure phellodone in an amount effective to reduce or inhibit protein carbonylation; b) cosmetically acceptable And c) optionally, i) one or more cosmetic ingredients selected from the group consisting of: alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosity modifiers, emulsifiers, stable Agents, colorants and fragrances; and/or ii) skin lightening agents selected from the group consisting of tyrosinase inhibitors, free radical scavengers, and mixtures thereof. A topical composition having reduced or inhibited protein carbonylation activity consisting of: a) at least about 90% pure paeonirone in an amount effective to reduce or inhibit protein carbonylation; b) a cosmetically acceptable vehicle; And c) optionally, i) one or more cosmetic ingredients selected from the group consisting of: alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosity modifiers, emulsifiers, stabilizers, colorants And a fragrance; and/or ii) a skin lightening agent selected from the group consisting of tyrosinase inhibitors, free radical scavengers, and mixtures thereof. The composition of any one of claims 1 to 2, wherein the corkone is purified from an extract of at least one plant selected from the group consisting of sage, white peony, peony bark, bergamot, medlar and toon . The composition of claim 3, wherein the extract of Evodia rutaecarpa is normalized to the content of at least one of evodiamine, rutaecarpine and limonin. The composition of any one of claims 1 to 4, wherein the cedarone is about 99.99% pure phellodone active. The composition according to any one of claims 1 to 5, which is a preparation selected from the group consisting of a cream, an ointment, a foaming agent, a lotion, a plaster, a tablet, a granule or an emulsion. The composition of any one of claims 1-2, wherein the ratio of the ketone to the skin lightening agent is from about 100:1 to about 1:100. The composition of any of claims 1-7, wherein the amount of protein carbonylation effective to reduce or inhibit is from about 10 to 1000 mg. The composition of any of claims 1-8, wherein the cedarone comprises from about 1 to 99% w/w of the composition. The composition of any of claims 1-8, wherein the cedarone comprises from about 1 to 75% w/w of the composition. The composition of any of claims 1-8, wherein the cedarone comprises about 50-70% w/w of the composition. A topical composition comprising a cosmetically acceptable vehicle and at least 90% pure paisone in an amount of from about 1 to 99% w/w of the composition. A method of improving the appearance of the skin or at least one sign of skin aging, The method comprises topically applying to the skin a cosmetically effective amount of a composition sufficient to improve the appearance of the skin or at least one sign of skin aging, wherein the composition comprises a. a quantity of about 75-100% pure parvonone; b. a cosmetically acceptable vehicle; and c. optionally, i) one or more cosmetic ingredients selected from the group consisting of: alcohols, fats, oils, surfactants, fatty acids, emu oils, humectants, humectants, viscosities Conditioning agents, emulsifiers, stabilizers, colorants and perfumes; and/or ii) skin lightening agents selected from the group consisting of tyrosinase inhibitors, free radical scavengers, and mixtures thereof. The method of claim 13, wherein the composition is a product selected from the group consisting of: aerosols, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, stickers Tablets, pomades, solutions, pump sprays, sticks, small towels and combinations thereof. A method of whitening the skin comprising topically applying the composition of any one of claims 1-12 to the skin, wherein the composition is applied in an amount and for a duration sufficient to visually whiten the skin. A method of brightening the skin comprising topically applying the composition of any one of claims 1-12 to the skin, wherein the composition is applied in an amount and for a duration sufficient to visibly brighten the skin. Use of a topical composition according to any one of claims 1 to 12 for the preparation of a cosmetic agent having activity to reduce or inhibit protein carbonylation. A method of preventing, ameliorating or reducing at least one sign of skin aging, It comprises applying the composition of any one of claims 1-12 to the skin, wherein the composition is applied in an amount and for a duration sufficient to prevent, ameliorate or reduce the at least one sign of skin aging.