HK1169949A - Compositions comprising a retinoid and an nfkb-inhibitor and their methods of use - Google Patents

Description

Compositions comprising retinoids and NFkB inhibitors and methods of use thereof

Technical Field

A composition comprising a retinoid, an NF κ B inhibitor and a cosmetically acceptable topical carrier is provided. The composition is useful for topical application to the skin.

Background

Retinoids are known to treat various skin conditions such as acne or photoaging. However, retinoids (such as retinoic acid, retinal or retinol) for topical application can in some cases lead to redness, itching, stinging, skin peeling or other signs of irritation. The present inventors have considered that one possible solution to reduce inflammation is to use lower concentrations of retinoids in topical compositions. However, it is understood that compositions with low levels of retinoids may have low efficacy.

The present inventors have now surprisingly found that a particular class of anti-inflammatory compounds, i.e. agents that inhibit the nuclear transcription factor k-B (nfkb), can be combined with retinoids in a manner that results in a surprisingly large, unexpected synergistic enhancement of retinol activity, as measured by expression of the CRAPBII gene.

Disclosure of Invention

In one aspect, the present invention provides a composition comprising a retinoid, an NF κ B inhibitor and a cosmetically acceptable topical carrier, wherein the amount of retinoid in the composition is no more than about 0.075% by weight of the composition.

In another aspect, the present invention provides a method of treating skin comprising topically applying the above composition to the skin of a mammal.

Other features and advantages of the invention will be apparent from the detailed description of the invention and from the claims.

Detailed Description

It is believed that one skilled in the art can, using the description herein, utilize the present invention to its fullest extent. The following specific examples are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Unless otherwise indicated, amounts, percentages, or concentrations refer to amounts, percentages, or concentrations by weight (i.e., (W/W)). Unless otherwise stated, all ranges are inclusive, e.g., "4 to 9" includes the endpoints 4 and 9.

As used herein, "topical application" means directly on or spread over the external skin, scalp or hair, for example, by hand or an applicator (e.g., a wipe, roller, or spray).

As used herein, "cosmetically acceptable" means that the ingredients described by this term are suitable for use in contact with tissue (e.g., skin or hair) without undue toxicity, incompatibility, instability, irritation, allergic response, and the like.

The term "treatment" or "treating" as used herein means treating (e.g., slowing or eliminating and/or curing of symptoms) and/or preventing or inhibiting a disorder (e.g., a skin disorder).

The compositions of the invention are suitable for treating mammalian skin, for example for improving various signs of skin aging, such as firmness, texture or appearance of wrinkles. As used herein, "skin in need of improvement of signs of aging" means, but is not limited to, sagging, loose, rough, wrinkled, thinned or uneven skin. Improving the signs of aging means improving skin firmness, improving skin texture, improving the appearance of fine lines or wrinkles in the skin, improving skin tone (tone), or treating external aggressions in the skin.

As used herein, "improving the firmness of skin" means enhancing the firmness or elasticity of skin, preventing loss of firmness or elasticity of skin or preventing or treating sagging, loose and flabby skin. The firmness or elasticity of the skin can be measured by using a skin elasticity tester (curometer). See Handbook of Non-Invasive Methods and the Skin, J.Serup, G.Jemec & G.Grove (eds.), Chapter 66.1 (2006). Loss of skin elasticity or firmness may be the result of a variety of factors including, but not limited to, aging, disease, hormonal changes, trauma, environmental damage, or the result of applying cosmetics to the skin.

As used herein, "improving the texture of the skin" means smoothing the skin surface to remove bumps or crevices on the skin surface.

As used herein, "ameliorating the appearance of fine lines or wrinkles in the skin" means preventing, delaying, stopping or reversing the process of wrinkle and fine line formation in the skin.

As used herein, "treating an external insult in the skin" means reducing or preventing damage from the external insult in the skin. Examples of external insults include, but are not limited to, the use of cleansers (e.g., topical cleansers containing surfactants), cosmetics, shaving, and environmental damage such as damage to the skin from UV light (e.g., sun damage from sunlight or damage from non-natural sources such as UV lamps and solar simulators), ozone, exhaust gases, pollution, chlorine and chlorine-containing compounds, and smoking. The effects of external aggressions on the skin include, but are not limited to, oxidative and/or nitrosative damage to and modification of lipids, carbohydrates, peptides, proteins, nucleic acids, and vitamins. The effects of external aggressions on the skin also include, but are not limited to, loss of cell viability, loss or alteration of cell function, and changes in gene and/or protein expression.

As used herein, the term "lightening skin" generally refers to lightening and/or depigmenting the skin tone, skin color, and/or skin shade (shade), lightening, whitening, and/or evening, and/or to removing yellow, and/or to lightening and/or depigmentation of pigmented spots and/or lesions (including, but not limited to, pigmented spots, melanin spots, age spots, sun spots, age spots, freckles, lentigo simplex, pigmented solar keratoses, seborrheic keratoses, melasma, acne spots, post-inflammatory hyperpigmentation, lentigo, combinations of two or more thereof, and the like). In certain embodiments, "whitening skin" also refers to increased skin radiance, translucency, and/or glow and/or to achieve a more radiant, translucent, or glowing skin tone appearance or a lower yellow or sallow skin tone. In certain preferred embodiments, "lightening skin" refers to lightening and evening out skin tone, increasing skin radiance, and/or lightening age spots.

As used herein, the term "skin in need of skin lightening treatment" generally refers to skin that exhibits one or more characteristics selected from the group consisting of: SKIN having Individual type horn (ITA) measurements OF less than 41 (measured according to THE COLIPA GUIDELINE: GUIDELINE FOR THE color pigment cosmetic OF SKIN color TYPING AND preliminary OF THE MINIMAL ERYTHEMAL dose (med) and UV extract OF SKIN described further below, which is incorporated herein by reference, and which is described further below), dark and/or sallow SKIN, including UV-darkened SKIN, SKIN having uneven SKIN tone, or SKIN having one or more pigmented spots and/or lesions (including but not limited to pigmented spots, melanin spots, age spots, sunburn, senile pigmented spots, freckles (freckles), lentigo lentigines, pigmented solar keratoses, melasma, post-acne pigmentation, hyperpigmentation, inflammation, freckles (e), and THE like, or a combination OF two or more OF these. In the COLIPA guidelines, skin color is defined according to ITA values as: very light-colored skin > 55; light skin 41-55, medium skin 28-41, and dark brown skin < 28. In certain preferred embodiments, "skin in need of skin lightening" refers to individuals having skin with an ITA value of less than 41, for example, about 40 or less, about 35 or less, about 30 or less, or more preferably about 28 or less. In certain preferred embodiments, the present invention relates to compositions and methods for use on skin in need of a skin lightening treatment selected from the group consisting of sallow and/or darkened skin. In certain other preferred embodiments, the present invention relates to compositions and methods for skin lightening treatments in need thereof selected from the group consisting of age spots, freckles, marks left after acne, and combinations of two or more thereof.

The compositions of the present invention are also suitable for treating acne. As used herein, "treating acne" refers to reducing, preventing, ameliorating, or eliminating the presence or signs of disorders caused by the action of hormones and other substances on the sebaceous glands and their hair follicles (which typically results in the blockage of pores and the formation of inflammatory or non-inflammatory lesions on the skin). In particular, it relates to the treatment or prevention of blemishes, lesions or pimples, pre-emergent pimples, blackheads and/or whiteheads. As used herein, a "pre-emergent pimple" is an inflamed hair follicle that is not evident (e.g., as a lesion) on the skin surface when viewed with the naked eye.

As used herein, "cosmetic" refers to a cosmetic substance or article that maintains, restores, imparts, stimulates or enhances the appearance of a body beauty or appears to enhance the appearance or youthfulness, particularly as it relates to the appearance of tissue or skin.

As used herein, "cosmetically effective amount" means an amount of a physiologically active compound or composition that is sufficient to treat one or more signs of skin aging or acne, but low enough to avoid serious side effects. The cosmetically effective amount of the compound or composition will vary depending on the following factors: the particular condition being treated, the age and physical condition of the end user, the severity of the condition being treated/prevented, the duration of the treatment, the nature of the other treatment, the particular compound or product/composition employed, the cosmetically acceptable carrier specifically employed, and the like.

Retinoids

The compositions of the present invention comprise at least one retinoid. As used herein, "retinoid" means a compound from a class of compounds that are structurally similar to vitamin a, such as those characterized by the following structure:

wherein R represents a functional group such as CH₂OH (retinol), CHO (retinal), CO₂H (Vision)Xanthic acid), CH₂OCOCH (retinol acetate). Other derivatives are also suitable, for example other esters such as retinol palmitate, amine derivatives and the like. In one embodiment, the retinoid is selected from the group consisting of retinol, retinal, retinoic acid, retinol acetate and retinol palmitate. In a preferred embodiment, the retinoid is retinol.

The inventors have found that relatively low levels of retinoid can be maintained in the composition, yet still provide surprisingly greater efficacy than expected. In certain embodiments of the invention, the amount of retinoid (e.g., retinol) in the composition is no more than about 0.075 weight percent of the composition (e.g., no more than about 0.0026 moles per liter of composition, or no more than about 247,500 international units of vitamin a per 100 grams of composition). In certain embodiments, the amount of retinoid in the composition is from about 0.01% to about 0.075% by weight of the composition (e.g., from about 0.000349 to about 0.0026 moles per liter of composition), such as from about 0.01% to about 0.06% by weight of the composition (e.g., from about 0.000349 to about 0.00209 moles per liter of composition). In a preferred embodiment, the composition comprises retinol in the above concentration ranges.

NF kappa B inhibitor

The compositions of the present invention comprise at least one NF κ B inhibitor. As used herein, "NF-. kappa.B inhibitor" means a compound that inhibits the nuclear transcription factor k-B (NF-. kappa.B) of a cellular transcription factor. In one embodiment, the nfkb inhibitor, when tested according to the nfkb inhibition test defined below, has a percent inhibition of nfkb of at least about 35%, preferably at least about 55%, more preferably at least about 70%, most preferably at least about 90%, when tested at a concentration of preferably 1 microgram per ml to about 100 micrograms per ml. That is, the compounds exhibit the listed percent inhibition of NF κ B at least one concentration in the range of 1 μ g per ml to 100 μ g per ml. The compounds need not provide the listed percent inhibition of nfkb at all concentrations from 1 mg per ml to 100 mg per ml, but at least one concentration within this range.

In a preferred embodiment, the nfkb inhibitor has a percent nfkb inhibition of at least about 35%, preferably at least about 55%, more preferably at least about 70%, most preferably at least about 90%, when tested at a concentration of 10 milligrams per milliliter.

The "NF κ B inhibition test" was performed in the following manner. FB293 cells, a stably transfected human epithelial cell line containing a reporter gene for NF-kB, were used. They are available, for example, from Panomics (Fremont, CA). FB293 at 5X 10⁴The cells/mL are seeded at a density in a suitable medium, for example Dartbox Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (Invitrogen, San Diego, CA). FB293 cells were stimulated with 100ng/mL tumor necrosis factor α (TNF α, available from Sigma-Alrich (St Louis, Mo)) in the presence of the test samples. Independently, a control sample was tested, wherein no test sample was applied. 5% CO at 37 deg.C₂After 24 hours of incubation, cells were lysed with 40 μ l of reporter lysis buffer (Promega, Madison, WI). Aliquots of 20 μ L of the lysate were measured using the luciferase assay kit (Promega) and counted for 10 seconds in an Lmax luminometer (Molecular Devices, Sunnyvale, Calif.), with data expressed in relative light units/second. The percent inhibition of NF κ B of the test samples was calculated as follows:

nfkb inhibition ═ 1- (L)_{Sample (I)}/L_Control)]*100

Wherein L is_{Sample (I)}Luminescence of the sample, L_ControlLuminescence for the control.

In one embodiment, the NF κ B inhibitor is selected from the following compounds: substituted resorcinols, (E) -3- (4-methylphenylsulfonyl) -2-acrylonitriles (such as "Bay 11-7082", commercially available from Sigma-Aldrich (st. louis, Missouri)), tetrahydrocurcumines (such as tetrahydrocurcumin CG, available from sabina Corporation (Piscataway, NJ)), extracts of Paulownia tomentosa (Paulownia tomentosa), and combinations thereof. In another embodiment, the NF κ B inhibitor is selected from substituted resorcinols, tetrahydrocurcuminoids, and combinations thereof.

In one embodiment, the NF κ B inhibitor comprises an extract of paulownia tomentosa. Paulownia (Paulownia) is a native genus of plants in asia that has gradually spread to europe and the united states. In japan, the genus paulownia is called kiri, which specifically refers to one species paulownia tomentosa, also called "princess tree". Other names commonly used are "queen tree", "digitalis tree", "royal paulownia", "paulownia" (chinese name) and "odd-Namoo" (korean name). The name of "Paulownia tomentosa" is a school name, and various aliases, namely, "Paulownia imperialis", "Paulownia recurva" and "Bignonia toneosa" are reported in various literatures. Paulownia tomentosa belongs to the family Paulowniaceae (Paulowniaceae) or sometimes referred to as the family Scrophulariaceae. The american department of agriculture (plantas. usda. gov) plant database identified queen trees by the unique symbol "PATO 2" with Paulownia fortunei and Paulownia imperialis as alias.

Any suitable extract of paulownia tomentosa may be used. Generally, the wood of paulownia tomentosa includes wood from a trunk, branches, or a combination of the two. Suitable extracts of paulownia tomentosa wood may be derived from wood chips, wood flour, and/or small cuttings, among others.

Suitable extracts of paulownia tomentosa may be obtained by conventional methods, including but not limited to: by grinding, macerating, squeezing, mashing, centrifuging, and/or by methods such as cold percolation, stirring/distillation, microwave assisted extraction, sonication, supercritical/subcritical CO with or without polar modifier₂Compressed gas extraction, pressurized solvent extraction, accelerated solvent extraction, pressurized or atmospheric hot water extraction, surfactant-assisted pressurized hot water extraction, oil extraction, membrane extraction, Soxhlet extraction, gold finger distillation/extraction (gold finger distillation/extraction) and/or processes and the like, such as disclosed in U.S. Pat. Nos. 7442391, 7473435, and 7537791, issued to Integrated cosmetic Technologies, LLC (incorporated herein by reference), and/or the likeOr by other methods such as solvent extraction or the like. Any of a variety of solvents, including polar solvents, non-polar solvents, or a combination of two or more thereof, can be used in a process that includes solvent extraction. Suitable polar solvents include: polar inorganic solvents such as water and the like, polar organic solvents such as monohydric alcohols and the corresponding organic acids (e.g., C including methanol, ethanol, propanol, butanol, and the like)₁-C₈Alcohols and organic acids including acetic acid, formic acid, propionic acid, etc.), polyols and glycols (including C)₁-C₈Polyols/glycols, etc.) and combinations of two or more thereof. Suitable non-polar solvents include non-polar organic solvents such as alkanes (including C)₁-C₈Alkanes), cycloalkanes (including C)₁-C₈Alkanes), alkyl ethers (including C)₁-C₈Alkyl ethers), petroleum ethers, ketones (including C)₁-C₈Ketones), methylene chloride, ethyl acetate, xylene, toluene, chloroform, vegetable oils, mineral oils, and the like. In another embodiment, polar modifiers such as (C) may be added by the above-mentioned nonpolar solvents or with or without the use of polar modifiers₁-C₈Alcohol, water, C₁-C₈Polyol/diol or C₁-C₈Organic acid) to obtain an extract. In certain preferred embodiments, the extract of the invention is a polar extract prepared by grinding wood into a powder and extracting with a polar solvent having a dielectric constant value between 1 and 100 at 20 ℃, preferably with a polar solvent having a dielectric constant value between 4 and 60 at 20 ℃, more preferably with a polar solvent having a dielectric constant value between 4 and 50 at 20 ℃, still more preferably with a polar solvent having a dielectric constant between 4 and 40 at 20 ℃. Examples of preferred polar solvents include C having a dielectric constant value of between 1 and 100, preferably between 4 and 60, more preferably between 5 and 40 at 20 deg.C₁-C₈Alcohol, C₁-C₈Polyol/diol, C₁-C₈Organic acids, Water, and combinations of two or more thereof, including but not limited to, for example, "Dielectric Constants of Some Organic Solvent-Water Mixtures atVariaus temperameres, "Akerlof, Gosta; JACS, Vol 54, No. 11 (11 months 1932), pp 4125-4139 (incorporated herein by reference) describes those solvents and combinations of solvents having the desired dielectric constant values. In certain preferred embodiments, one or more C is used₁-C₈Monohydric alcohol, C₁-C₈Polyol, C₁-C₈Glycols and combinations of two or more thereof. In certain more preferred embodiments, one or more C is used₁-C₄Monohydric alcohol, C₁-C₄Polyol and/or C₁-C₄Extracting the extract with glycol. In certain more preferred embodiments, the extract is prepared with a solvent comprising methanol, ethanol, or a combination thereof, in the presence or absence of water. In a more preferred embodiment, the extract is prepared with dry alcohol or reagent grade denatured alcohol and dry Kiri wood powder, stirred at room temperature for 3 days. In certain preferred embodiments, the extract may be further refined by charcoal (also known as activated charcoal).

In certain embodiments, the paulownia tomentosa extract may be prepared to be substantially free of certain substances. In one embodiment, the extract is substantially free of ursolic acid, beta-sitosterol, or both.

In certain embodiments, the composition may additionally comprise an extract from other parts of paulownia tomentosa (e.g., one or more of the bark, leaves, roots, fruits, seeds, or flowers). In other embodiments, the composition is substantially free of extracts from other non-woody parts of paulownia tomentosa.

In a preferred embodiment, the NF κ B inhibitor is a substituted resorcinol. Resorcinol is a dihydric phenol compound (i.e., 1, 3 dihydroxybenzene) having the structure:

as used herein, "substituted resorcinol" means resorcinol having at least one substituent in the 2, 4, 5 or 6 position. Thus, a substituted resorcinol may have as few as one and as many as four substituents. The positions 1 and 3 of the substituted resorcinols have an-OH hydroxyl group, as shown above.

It is highly preferred that all substituents of the substituted resorcinol are free of phenyl (-C6H5 aromatic) moieties. In certain embodiments, all substituents are free of aromatic moieties (with or without heteroatoms).

In another embodiment, it is preferred that all substituents of the substituted resorcinol be free of ketone functionality (carbonyl groups bonded to two other carbon atoms).

In certain preferred embodiments, all substituents of the substituted resorcinol have neither a phenyl nor a ketone functionality.

In certain preferred embodiments, the substituted resorcinol comprises at least one substituent having from 5 to 11 carbon atoms, preferably from 5 to 10 carbon atoms, more preferably from 5 to 9 carbon atoms, and most preferably from 5 to 8 carbon atoms. In certain other embodiments, at least one substituent comprises an alkyl group, such as an alkyl group having the above-described number of carbon atoms. The alkyl group is preferably unsaturated.

In certain embodiments, the 4 position of resorcinol is substituted, and in certain embodiments, only the 4 position is substituted. In another embodiment, the 4 position is substituted with an alkyl group. In certain preferred embodiments, the substituted resorcinol comprises a single substituent at the 4-position, which substituent comprises an alkyl group. In certain other preferred embodiments, the substituted resorcinol contains a single 4-position substituent consisting of an alkyl group bonded directly to the phenyl ring.

Particularly suitable substituted resorcinols include 4-hexylresorcinol and 4-octylresorcinol, especially 4-hexylresorcinol. The structures of 4-hexylresorcinol and 4-octylresorcinol are shown below:

4-hexylresorcinol

4-octyl resorcinol

4-hexylresorcinol is available from Sytheon (Lincoln Park, NJ) under the trade designation "SYNOVEA HR". 4-Octylresorcinol is available from City Chemical LLC (West Haven, Connecticut).

In certain embodiments, the substituted resorcinol has at least two substituents at the 2, 4, 5, or 6 positions. Such substituents may optionally be linked to form a ring, for example a cyclic aliphatic hydrocarbon optionally containing heteroatoms such as sulphur or oxygen. For example, the attached substituent may contain 5 to 10 carbon atoms, such as 8 to 10 carbon atoms, and optionally include 1 to 3 heteroatoms. Examples of suitable substituted resorcinols having a cyclic aliphatic substituent linking the 2 and 3 positions include zearalenone and β -zearalanol:

zearalenone (zearalenone)

Beta-zearalanol

Zearalenone and β -zearalanol are commercially available from Sigma Chemicals (st. louis, Missouri).

In certain other embodiments, the substituted resorcinol has halogen-containing and/or nitroso-containing substituents. Suitable examples contain-Cl or-N ═ O bonded directly to the phenyl ring. These substituents may be present, for example, in the 2 and 4 positions, 2 and 6 positions or 4 and 6 positions. An example of a dihalo-substituted resorcinol is 2, 6-chlororesorcinol. Examples of dinitroso-substituted resorcinols are 2, 4-dinitroso-resorcinols:

2, 4-dinitrosorcinol

2, 6-dichlororesorcinol and 2, 4-dinitrosorcinol are available from City Chemical LLC (West Haven, Connecticut).

Substituted resorcinols can be prepared by methods known in the art, for example, using the techniques described in U.S. patent No.4,337,370, the contents of which are incorporated herein by reference.

The substituted resorcinol can have any suitable molecular weight. In certain embodiments, the molecular weight of the substituted resorcinol ranges between about 175 to about 300.

CRABBII expression

In one embodiment of the invention, the concentration of the NF κ B inhibitor in the composition is selected to provide at least about a 15% potentiation (in combination with a retinoid) of CRAPBII expression as measured by the "CRAPBII expression test" described below. This combination advantageously enables the use of relatively low levels of retinoid (which is sometimes irritating) while maintaining a high degree of retinoid activity. In humans, the CPABPII gene encodes for the synthesis of cellular retinoic acid binding proteins. As such, the expression of the CRABPII gene is directly related to and strongly correlated with retinoid efficacy. The present inventors have found that the presence of NF κ B inhibitors in a composition containing a retinoid may improve or potentiate the beneficial effects of the retinoid, as measured by expression of CRAPB II.

The degree of potentiation ("% potentiation") shown by CRAPBII expression for a particular combination of retinoid and NF κ B inhibitor can be determined using the "CRAPBII expression test". "CRABBII expression assayThe test "was performed in the following manner. One centimeter diameter ex vivo skin explants were prepared from skin abdominal biopsy samples after plastic surgery. Skin explants were maintained at 37 ℃ in a water-saturated atmosphere at KGM gold supplemented with 0.125. mu.g/ml amphotericin B^TMIn the medium, the duration of the test was 48 hours. The explants are placed in a conventional test plate with the epidermal surface facing up and in sufficient medium to almost, but not completely, submerge the sample (i.e., the epidermal surface protrudes from the upper surface of the medium). The test composition is applied to the epidermal surface protruding from the culture medium. After 48 hours, the explants are removed, epidermal mRNA is extracted therefrom, and CPABPII gene expression is measured by quantitative real-time PCR (QRT-PCR) using appropriate oligonucleotide sequences, using conventional techniques known to those skilled in the art.

A water-in-oil emulsion base containing the desired retinoid was used in the test compositions. The nfkb inhibitor may optionally be included in the test composition or separately mixed into the culture medium. For example, depending on the relative permeation rate of the NF κ B inhibitor, it may be advantageous to include it in the test composition or culture medium to eliminate the permeation rate as a variable factor in the test. If contained in the medium, the NF-. kappa.B inhibitor need not penetrate into the epidermis from the surface of the protruding epidermis, but may penetrate into the epidermis from the underlying medium. If an nfkb inhibitor is included in the test composition, the nfkb inhibitor must penetrate into the epidermis from the protruding epidermal surface.

The concentration of water in the test composition can be adjusted to accommodate a sufficient level of retinol (and the NF κ B inhibitor, if present in the test composition). Examples of oil-in-water emulsion bases that can be used in the test compositions are shown in table 1 below:

table 1: suitable oil-in-water emulsion matrices for CRABBII expression testing

Trade name	INCI	Concentration (percentage by weight)
			Demineralized water	Water (W)	83.4
Carbopol Ultrez 10	Carbomer	0.4
			Edeta BD chelating agent	Ethylenediaminetetraacetic acid disodium salt	0.1
Nipagin M	P-hydroxybenzoic acid methyl ester	0.2
			Propyl p-hydroxybenzoate	Propyl p-hydroxybenzoate	0.15
Phenoxyethanol	Phenoxyethanol	0.5
			Sodium hydroxide	Sodium hydroxide	0.1
Simulsol 165	PEG-100 stearate; stearic acid glyceride	2.0
			Nipanox BHT	BHT	0.1
Lanette 16	Cetyl alcohol; stearyl alcohol; tetradecanol	1.0
			DUB ININ	Isononyl isononanoate	7.0
Propylene glycol	Propylene glycol	5.0
			Ascorbic acid crystal	Ascorbic acid	0.05

[0067](i) CRAPBII expression was determined for a given amount of retinoid alone, (ii) for a given amount of NF κ B inhibitor alone, and (iii) for a mixture of amount (i) of retinoid and amount (ii) of NF κ B inhibitor. The% synergy was calculated using the formula:

100% × [ [ CRABPII ] yield_{Mixture of}/(CRAPBII_Retinoids+CRAPBII_{NF kappa B inhibitor})]-1]

While the concentration of the nfkb inhibitor may be selected to provide at least about 15% synergistic CRABPII expression for the compositions of the invention, in certain embodiments, the level of the nfkb inhibitor may be selected to provide a higher level of synergy, e.g., at least about 50%, or at least about 80% synergy, in CRABPII expression.

In one embodiment, the concentration of the nfkb inhibitor can be selected, for example, using the results of a "nfkb inhibition test". For example, suitable concentrations can be estimated by conducting the "nfkb inhibition test" at several (e.g., 4 or more) concentrations, as described in example 1 below. The concentration of the nfkb inhibitor can be selected, for example, such that a percent NF-kB inhibition of at least about 35%, preferably at least about 55%, more preferably at least about 70%, and most preferably at least about 90% is provided when tested at a concentration of 10 μ g/ml. This concentration of the nfkb inhibitor can be combined with no more than about 0.075% by weight of a retinoid in a composition and the% potentiation of CRAPBII expression determined as shown in example 2 below.

The inventors have observed that, in certain embodiments, a potentiation of CRAPBII expression is surprisingly observed not only at low concentrations of retinoid (not more than about 0.075 wt%), but also at low concentrations of NF κ B inhibitor. This is advantageous because the concentration of the retinoid can be maintained at a low level, thereby avoiding possible negative stimulatory side effects of the retinoid, while at the same time obtaining a potent efficacy enhancement from the NF κ B inhibitor.

In certain embodiments of the invention, the composition comprises no more than about 0.075% by weight of a retinoid, e.g., retinol, and no more than about 0.5% by weight of an NF κ B inhibitor. In a preferred embodiment, the composition comprises no more than about 0.075% of a retinoid and from about 0.1% to about 0.5% of an nfkb inhibitor.

Topical compositions

The compositions of the present invention may be topically applied to human skin and/or hair. In addition to the NF κ B inhibitor and the retinoid, the composition further comprises a cosmetically acceptable topical carrier, which may be about 50% to about 99.99% by weight of the composition (e.g., about 80% to about 99.9% by weight of the composition). In a preferred embodiment of the invention, the cosmetically acceptable topical carrier is or comprises water. The cosmetically acceptable topical carrier may include one or more ingredients selected from humectants, emollients, oils, humectants, and the like. In one embodiment, the cosmetically acceptable topical carrier comprises a substrate such as a nonwoven fabric or film material.

The compositions may be prepared in a variety of product types including, but not limited to, lotions, creams, gels, sticks, sprays, ointments, cleansing liquid lotions and solid soaps, shampoos, pastes, foams, powders, mousses, shaving creams, wipes, patches, hydrogels, film-forming products, masks and skin films, and cosmetics such as foundations, and mascaras. These product types may contain several types of cosmetically acceptable topical carriers including, but not limited to, solutions, suspensions, emulsions (e.g., microemulsions and nanoemulsions), gels, solids, and liposomes.

The compositions useful in the present invention can be formulated as solutions. Solutions typically comprise an aqueous or organic solvent (e.g., from about 50% to about 99.99% or from about 90% to about 99% of a cosmetically acceptable aqueous or organic solvent). Examples of suitable organic solvents include: propylene glycol, polyethylene glycol (200-.

The compositions useful in the subject invention may be formulated as a solution comprising an emollient. Such compositions preferably comprise from about 2% to about 50% of one or more emollients. As used herein, "emollient" refers to a material used to prevent or alleviate dryness, e.g., by preventing transepidermal water loss from the skin. Examples of emollients include, but are not limited to, vegetable oils, mineral oils, aliphatic esters, and the like.

Lotions may be made from such solutions. Lotions typically comprise from about 1% to about 20% (e.g., from about 5% to about 10%) of one or more emollients and from about 50% to about 90% (e.g., from about 60% to about 80%) moisture.

Another product that can be formulated from a solution is a cream. A cream typically comprises from about 5% to about 50% (e.g., from about 10% to about 20%) of one or more emollients and from about 45% to about 85% (e.g., from about 50% to about 75%) moisture.

Although it is preferred that the compositions of the present invention include water, the compositions may alternatively be anhydrous or a paste which is free of water but contains organic and/or silicone solvents, oils, lipids and waxes. Ointments may comprise a simple animal or vegetable oil base or a semi-solid hydrocarbon. The cream may comprise from about 2% to about 10% of one or more emollients and from about 0.1% to about 2% of one or more thickeners.

The composition may be formulated as an emulsion. If the topical carrier is an emulsion, from about 1% to about 10% (e.g., from about 2% to about 5%) of the topical carrier comprises an emulsifier. The emulsifier may be nonionic, anionic or cationic. Examples of suitable emulsifiers include those generally identified as suitable emulsifiers in the personal care and cosmetic formulation arts.

Lotions and ointments may be formulated as creams. Typically such emulsions comprise from 0.5% to about 5% of an emulsifier. Such creams typically contain from about 1% to about 20% (such as from about 5% to about 10%) of one or more emollients; from about 20% to about 80% (e.g., from 30% to about 70%) water; and from about 1% to about 10% (such as from about 2% to about 5%) of one or more emulsifiers.

Oil-in-water and water-in-oil single emulsion skin care formulations, such as lotions and ointments, are well known in the cosmetic arts and may be used in the subject invention. Multiple emulsion compositions (e.g., water-in-oil-in-water or oil-in-water-in-oil) may also be used in the subject invention. Typically, such single or multiple phase emulsions contain water, emollients, and emulsifiers as their essential ingredients.

The compositions of the present invention may also be formulated as a gel (e.g., an aqueous, alcoholic/aqueous, or oily gel made using a suitable gelling agent). Suitable gelling agents for aqueous and/or alcoholic gels include, but are not limited to, natural gums, acrylic acid and acrylate polymers and copolymers, and cellulose derivatives (e.g., hydroxymethyl cellulose and hydroxypropyl cellulose). Suitable gelling agents for oils (e.g., mineral oils) include, but are not limited to, hydrogenated butylene/ethylene/styrene copolymers and hydrogenated ethylene/propylene/styrene copolymers. Such gels typically comprise between about 0.1 and 5 wt% of such gelling agents.

The compositions of the present invention may also be formulated as solid formulations (e.g., wax-based sticks, bar compositions, powders, or wipes containing powders).

In addition to the foregoing components, the compositions used in the subject invention may also contain a wide variety of other oil-soluble and/or water-soluble materials commonly used in compositions for application to the skin and hair at levels established in the art.

Additional cosmetic active agents

In one embodiment, the composition further comprises another cosmetically active agent as used herein, a "cosmetically active agent" is a compound (e.g., a synthetic compound or a compound isolated from a natural source or natural extract) that has a cosmetic or therapeutic effect on skin or hair, including, but not limited to, anti-acne agents, oil control agents, antimicrobial agents (non-NF κ B inhibitors), antifungal agents, antiparasitic agents, external analgesics, sunscreens, photoprotective agents, antioxidants, keratolytic agents, surfactants, moisturizers, nutrients, vitamins, energy enhancers, antiperspirants, astringents, firming agents, anti-callus agents (anti-callous agents), and agents for hair and/or skin conditioning.

In one embodiment, the agent is selected from, but not limited to, hydroxy acids, benzoyl peroxide, D-panthenol, octyl methoxycinnamate, titanium dioxide, octyl salicylate, homosalate, avobenzone, carotenoids, radical scavengers, spin traps, amines (e.g., neutrol), ceramides, polyunsaturated fatty acids, essential fatty acids, enzymes, enzyme inhibitors, minerals, hormones such as estrogens, steroids such as hydrocortisone, 2-dimethylaminoethanol, copper salts such as copper chloride, copper-containing peptides such as Cu: Gly-His-Lys, coenzyme Q10, peptides, amino acids such as proline, vitamins, lactobionic acid, acetyl coenzyme A, niacin, riboflavin, thiamine, ribose, electron transfer substances such as NADH and FADH2, and electron transfer substances such as aloe vera, feverfew, Other plant extracts such as oatmeal and derivatives and mixtures thereof. The cosmetically active agent will typically be present in the compositions of this invention in an amount of from about 0.001% to about 20% (e.g., from about 0.005% to about 10%, e.g., from about 0.01% to about 5%) by weight of the composition.

Examples of vitamins include, but are not limited to, vitamin B (e.g., vitamin B3, vitamin B5, and vitamin B12), vitamin C, different forms of vitamin K and vitamin E (e.g., alpha, beta, gamma, or delta tocopherols), or mixtures and derivatives thereof.

Examples of hydroxy acids include, but are not limited to, glycolic acid, lactic acid, malic acid, salicylic acid, citric acid, and tartaric acid.

Examples of antioxidants include, but are not limited to, water-soluble antioxidants such as sulfhydryl compounds and their derivatives (e.g., sodium metabisulfite and N-acetyl-cysteine), lipoic acid and dihydrolipoic acid, resveratrol, lactoferrin, and ascorbic acid derivatives (e.g., ascorbyl palmitate and ascorbyl polypeptide). Oil-soluble antioxidants suitable for use in the compositions of the present invention include, but are not limited to, butylated hydroxytoluene, tocopherols (e.g., tocopherol acetate), tocotrienols, and ubiquinone. Natural extracts containing antioxidants suitable for use in the compositions of the present invention include, but are not limited to, extracts containing flavonoids and isoflavones and their derivatives (e.g., genistein and bigeminy zein), extracts containing resveratrol, and the like. Examples of such natural extracts include grape seed, green tea, pine bark and propolis.

Other materials

Various other materials may also be present in the composition, as is well known in the art. This includes humectants, pH adjusters, chelating agents (e.g., EDTA), fragrances, dyes, and preservatives (e.g., parabens) such as those commonly used in cosmetic formulations.

Compositions, formulations, and products comprising such compositions of the invention can be prepared using methods known to those of ordinary skill in the art.

Application method

The compositions of the present invention may be topically applied to mammalian skin in need of treatment to ameliorate one or more signs of skin aging or acne as described above. In one embodiment, the composition may be applied to skin in need of improvement in firmness, texture, or appearance of fine lines and wrinkles. The composition may be applied to skin in need of such treatment according to a suitable treatment regimen, for example, from up to twice a day to as little as once every three days or so.

In certain embodiments, the compositions of the present invention may also be used to treat other desired conditions associated with the skin. For example, the compositions of the present invention may be used to treat post-inflammatory hyperpigmentation, to reduce pore size, to reduce sebum production, and to mitigate scarring. In certain other embodiments, the compositions of the present invention may be administered simultaneously with or within hours of a mechanical or physical exfoliation treatment (e.g., microdermabrasion treatment), or with a chemical exfoliant or keratolytic agent such as salicylic acid. In certain other embodiments, the compositions of the present invention may be applied to minor wounds or post-operative sites to aid in healing.

It is believed that one skilled in the art can, using the description herein, utilize the present invention to its fullest extent. The following specific examples are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. The following non-limiting examples further illustrate the invention.

Example 1: NF kappa B inhibition

The above-described "NF κ B inhibition test" was performed on the following test samples: bay11-7082 test samples (Sigma-Aldrich, St. Louis, MO), tetrahydrocurcumin CG (Sabinsa Corporation, Piscataway, NJ), and various concentrations of 4-hexylresorcinol. The results are shown in table 2, where NF-kB reporter activation (luminescence, L) was recorded for the test and control samples. Percent NF-kB inhibition was also recorded.

Table 2: results of the NF κ B inhibition assay

Bay11-7082, tetrahydrocurcumin CG and 4-hexylresorcinol showed strong NF-. kappa.B inhibition.

Example 2: expression of 4-hexylresorcinol and retinol-CRABPII

A series of test compositions containing retinol, 4-hexylresorcinol, or a combination of the two compounds were subjected to CRAPBII expression testing as described above. For this series of tests, the oil-in-water emulsion base described in table 1 was used. Retinol and/or 4-hexylresorcinol are included in the water-in-oil emulsion matrix. Values for CRAPBII expression are reported in table 3 below. For the combinations, the% synergy calculated as described above was also recorded.

Table 3: CRABBII expression

As shown in table 3, CRAPBII expression is positively correlated with and quite sensitive to retinol concentrations within the range tested. 4-hexylresorcinol used in low amounts generally improved CRABBII expression, while higher amounts of 4-hexylresorcinol generally reduced CRABBII expression. A synergistic response is surprisingly observed at retinol concentrations not exceeding about 0.075 wt% of the composition. Furthermore, the synergy is greatly amplified when the concentration of 4-hexylresorcinol is about 0.5 wt% (about 50 μ g/ml).

Example 3: expression of Bay11-7082 and retinol-CRABPII

A series of test compositions containing retinol, Bay11-7082, or a combination of these two compounds were tested for CRABBII expression. For this series of tests, the oil-in-water emulsion base shown in table 4 below was used to make up the balance of the test composition.

Table 4: oil-in-water emulsion base

Composition (I)	Concentration (percentage by weight)
		Water (W)	55.7123
Acryloyldimethyl ammonium taurate/VP	0.5
		Ethylenediaminetetraacetic acidDisodium salt	0.1
Glycerol	5.0
		Butanediol	2.0
PEG-8	5.0
		Cetyl alcohol: ceteareth-20	3.0
Stearyl alcohol; ceteareth-20	3.0
		Ethylhexyl methoxycinnamate	2.0
Isohexadecane	1.5
		PPG-15 stearyl ether; BHT (butylated Hydroxytoluene)	4.5
Pentaerythritol tetra (ethylhexanoate)	7.0
		Shea butter (shea butter)	1.0
Tocopherol acetate	0.25
		BHT	0.1
Polydimethylsiloxane	2.0
		Cyclohexasiloxane; cyclopentasiloxane	2.01
Polyacrylamide; c13-14 isoparaffin; laureth-7	2.0
		Nylon-12	3.0
Ascorbic acid	0.05
		Citric acid	0.25
Sodium hydroxide	0.0277

[0114]For this series of tests, retinol was included in the test compositions. However, Bay11-7082 was not contained in a water-in-oil emulsion matrix, but was contained in the culture medium. CRABBII expression is recorded in Table 5 below. For the combination, the% synergy was also recorded.

Table 5: CRABBII expression

As shown in Table 5, a synergistic CRABBII expression was found for retinol in combination with the NF-kB inhibitor Bay 11-7082. Although the combination of 0.04% retinol with 1 μ M BAY11-7082 showed synergistic CRABBII expression, no synergy was observed for the combination of 0.04% retinol with 10 μ M Bay 11-7082.

The data demonstrate that the combination of an NF-KB inhibitor and a retinoid in a composition wherein the retinoid is present at a concentration not greater than about 0.075% by weight of the composition produces a surprising, synergistic increase in retinol activity.

Claims (17)

1. A composition comprising a retinoid, an nfkb inhibitor, and a cosmetically acceptable topical carrier, wherein the amount of retinoid in the composition is no more than about 0.075% by weight of the composition.

2. The composition of claim 1, wherein the nfkb inhibitor is selected from the group consisting of substituted resorcinols, tetrahydrocurcumin, (E) -3- (4-methylbenzenesulfonyl) -2-acrylonitrile, extracts of paulownia tomentosa, and combinations thereof.

3. The composition of claim 1, wherein the nfkb inhibitor is selected from the group consisting of substituted resorcinols, (E) -3- (4-methylbenzenesulfonyl) -2-acrylonitriles and combinations thereof.

4. The composition of claim 1, wherein the nfkb inhibitor is selected from the group consisting of substituted resorcinols, tetrahydrocurcuminoids, and combinations thereof.

5. The composition of claim 1, wherein the nfkb inhibitor is a substituted resorcinol.

6. The composition of claim 1, wherein the nfkb inhibitor is 4-hexylresorcinol.

7. The composition of claim 6 comprising no more than about 0.5% by weight of 4-hexylresorcinol.

8. The composition of claim 6, comprising from about 0.1% to about 0.5% by weight of 4-hexylresorcinol.

9. The composition of claim 1, wherein the retinoid is retinol.

10. The composition of claim 9 comprising from 0.01% to about 0.075% by weight retinol.

11. The composition of claim 9, comprising 0.01% to about 0.06% by weight retinol.

12. The composition of claim 1, wherein said retinoid and said nfkb inhibitor are present in amounts sufficient to provide a percent potentiation of CRABPII activation of at least about 15%.

13. A method of treating mammalian skin comprising applying the composition of claim 1 to the skin.

14. The method of claim 13, wherein the skin is in need of improvement in the signs of skin aging.

15. The method of claim 14, wherein the skin is in need of improvement in firmness, texture, appearance of fine lines or wrinkles, or loss of elasticity.

16. The method of claim 13, wherein the skin is in need of improvement in signs of acne.

17. The method of claim 16, wherein said skin comprises blemishes, lesions, or papules, pre-emergent papules, blackheads, and/or whiteheads.