MXPA00008982A - Cosmetic method of treating skin - Google Patents

Abstract

A cosmetic method for treating aged, wrinkled and/or photodamaged skin is provided through topical application of a composition which comprises petroselinic acid and/or derivatives thereof. The method also reduces skin irritation and also lightens the colour of skin.

Description

COSMETIC METHOD FOR TREATING SKIN This invention relates to a cosmetic method for improving the condition and appearance of the skin and the use of petroselic acid in the preparation of topical compositions to improve the condition and appearance of the skin. The skin is subject to deterioration as a consequence of dermatological disorders, environmental abuse (wind, air conditioning, central heating) or as a consequence of the normal process of aging (chronological aging) that can be accelerated by the exposure of the skin to the sun (aging by effects of light). In recent years, the demand for cosmetic compositions and cosmetic methods to improve the appearance and condition of the skin has grown in large proportions. Increasingly, consumers are increasingly looking for "anti-aging" cosmetic products that treat or retard the visible signs of chronological aging and aging by effects of light on the skin such as wrinkles, fine lines, sagging, hyperpigmetation and age spots. Also, consumers often seek other benefits of cosmetic products in addition to the effect "against aging". The concept of "sensitive skin" has also raised the demand of the consumer for cosmetic products that improve the appearance and condition of a sensitive skin, vedijuda and / or dry, and to relieve red and / or irritated skin. Consumers also want cosmetic products that treat stains, pimples and defects, etc. Many people are concerned about the degree of pigmentation of their skin. For example, skin with age spots or freckles want these pigmentation spots to be less noticeable. Others want to reduce the darkening of the skin caused by sun exposure or wish to lighten their natural skin color. To meet these needs, many attempts have been made to develop products that reduce pigment production in melanocytes. However, the substances hitherto identified tend to have undesirable side effects, for example, skin irritation. Consequently, such substances are not suitable for cosmetic use and can only be applied in concentrations in which their lightening effect on the skin is less desired. Using a combination of different skin lightening substances can be considered to reduce side effects, but there is a potential risk that using this combination will reduce the skin lightening effect due to the effects of competition. Therefore, there is a need to improve the effectiveness of skin lightening cosmetic products, particularly those that do not irritate the skin. The use of fatty acids, among which petrosellenic acid is counted, is well known for cosmetic formulations for the treatment of hair. EP-A-116439) discloses hair tonics that include fatty acids (such as petrosellenic acid) to relieve dandruff and stinging itching and to stimulate hair growth. EP-A-709084 discloses the use of a coriander seed oil, which is rich in petrosellenic acid triglyceride, in a cosmetic composition for skin wetting conditions. Now, we have found other undiscovered properties of petrosellenic acid, which are useful in cosmetic compositions for topical application to the skin to provide skin care benefits not previously discovered. It has been found that an effective treatment and the prevention of normal skin conditions due to chronological aging or aging due to light effects, such as wrinkles, expression lines, sagging, hyperpigmentation and age spots, can be obtained through the application of cosmetic compositions on the skin which comprise petrosellenic acid or derivatives thereof. Also, it has been found that the use of petrosellenic acid in cosmetic compositions advantageously provides other benefits for the skin as well as anti-aging benefit such as relieving irritated and / or sensitive skin and rinsing of the skin.

BRIEF DESCRIPTION OF THE INVENTION In accordance with a first aspect of the present invention there is provided a cosmetic method for providing at least one skin care benefit selected from: treatment / prevention of wrinkles, sagging, aged and / or damaged skin by effects of light; high deposition of collagen in the skin, high production of decorin in the skin, improvement in tissue recovery; rinsing of the skin; relieve irritations, red and / or sensitive skin; improve the texture of the skin; softness and / or firmness; The method comprises applying to the skin a topical composition comprising petrosellenic acid and / or derivatives thereof. The present invention also encompasses the use of petrosellenic acid and / or derivatives thereof in a topical composition to provide at least one skin care benefit selected from: treatment / prevention of wrinkles, sagging, aging skin and / or damaged by effects of light; high deposition of collagen in the skin, high production of decorin in the skin, improvement in tissue recovery; rinsing of the skin; relieve irritations, red and / or sensitive skin; improvement of skin texture; softness and / or firmness. The inventive methods and use of petrosellenic acid provides anti-aging benefits, which result in the promotion of a soft and supple skin with improved elasticity and a reduced or delayed appearance of wrinkles and aging skin, with a color of improved skin A general improvement in appearance, texture and condition is achieved, in particular with respect to a radiant, clear appearance and a youthful overall appearance of the skin. The methods and inventive uses are also beneficial in relieving and soothing irritated and / or sensitive skin. Petrosellenic acid is also useful for topical application in human skin to reduce the production of melanin and thereby lighten the skin on which it was applied. In this way, the inventive methods advantageously provide a wide range of benefits for skin care. The term "treatment" as used herein, within its scope, includes reducing, retarding and / or avoiding the aforementioned skin conditions such as wrinkled, aged, damaged by light and / or irritated skin and it generally improves the quality of the skin and improves its appearance and texture by avoiding or reducing wrinkles and increasing its flexibility, firmness, softness, flexibility and elasticity for the skin. The cosmetic methods and the uses of petrosellenic acid invention, can be useful for a skin in treatment, which already presents wrinkles, aging, or characteristics of irritation or damage by effects of light or for the treatment of young skin to thus avoid or reduce the aforementioned deterioration changes due to aging process by normal age / light.

DETAILED DESCRIPTION OF THE INVENTION Petrosellenic acid, hereinafter called PA is a long chain (C18) monounsaturated fatty acid, with a formula CH3 (CH2) 10CH = CH (CH2) 4COOH. The invention also includes the free acid derivatives which thus comprise petrosellenic acid fractions. Preferred derivatives include substitution derivatives of the carboxyl group of the acid, such as esters (for example, retinyl esters, triglyceride esters, monoglyceride esters, diglyceride esters, phosphoesters), amides (eg, ceramide derivatives), salts ( for example, alkali metal, alkali earth metal salts, ammonia salts); and / or those derived from the substitution of the C18 carbon chain, such as alpha hydroxy and / or beta hydroxy derivatives. In the case of triglyceride ester derivatives, all positional isomers of PA substitutes are included in the basic structure of glycerol. Triglycerides must contain at least a fraction of PA. For example, of the three esterifiable positions in the basic structure of glycerol, positions 1 and 2 can be esterified with PA and with another lipid in position 3, or alternatively, the basic structure of glycerol can be esterified with PA in positions 1 and 3 with another lipid at position 2. Oils that are rich in petrosellenic acid triglyceride are also, thus, suitable for use in the present invention. These oils are available for sale and include parsley seed oil, carrot seed oil, fennel fruit oil, parsnip seed oil, coriander seed oil, perifolio seed oil, caraway plant oil, and celery seed oil. Where the term "petrosellenic acid" or "PA" is used within this specification it should be understood that derivatives thereof comprising fractions of PA are also included. The "PA fractions" refer to a portion (portions) of fatty acyl of a PA derivative. The petroselic acid, active to be employed in accordance with the present invention is present in the topical composition in an effective amount. Normally, the total amount of active is present in an amount between 0.0001% to 50% by weight of the composition. More preferably, the amount is from 0.0% to 10% and with greater preference is from 0.1% to 5% in order to obtain the maximum benefits at a minimal cost. Acceptable Dermatological Vehicle The composition used according to the invention also comprises a dermatologically / cosmetically acceptable vehicle to act as a diluent, dispersant or carrier for the active PA. The vehicle may comprise materials commonly used in skin care products such as water, liquid or solid emollients, silicone oils, emulsifiers, solvents, humectants, thickeners, powders, propellants and their like. Usually, the vehicle will form from 5% to 99.9%, preferably from 25% to 80% by weight of the composition, and can, in the absence of other cosmetic adjuncts, form the balance of the composition. Optional Beneficial Materials for the Skin v Attachments Cosmetics In addition to the active ingredient, PA, other specific assets beneficial to the skin, such as sun blockers, other skin lightening agents, skin tanning agents, can also be included. The vehicle may also include adjuncts such as perfumes, opacifiers, preservatives, dyes and buffers.

Preparation of the Product, Form. Use v Packaging To prepare the topical composition used in the method of the present invention, the usual way can be used to prepare skin care products. The active components are generally incorporated in a dermatologically acceptable vehicle in a conventional manner. The active components can first be adequately dissolved or dispersed in a portion of water or other solvent or liquid to be incorporated into the composition. Preferred compositions are oil-in-water or water-in-oil emulsions. The composition may be in the form of conventional skin care products such as cream, gel, lotion or the like thereof. The composition may also be in the form of so-called "washable products", for example, a bath or a bath gel, which possibly contains a delivery system for the active ingredients to promote adhesion to the skin during rinsing. More preferably, the product is a "permanence" product; a product that is applied to the skin without having the step of deliberate rinsing after applying it on the skin. The composition can be packaged in any suitable manner such as a jar, bottle, tube, spinning ball, or the like in a conventional manner. The method of the present invention can be applied one or more times daily on the skin that requires treatment. Improvements in the appearance of the skin will usually be visible after 3 to 6 months, depending on the condition of the skin, the concentration of the active compound components used in the inventive method, the amount of composition used and the frequency with which applies In general, a small amount of the composition, for example 0.1 to 5 ml, is applied to the skin from the appropriate container or applicator and is distributed over and / or rubbed on the skin using the hands and fingers or any other proper device. Optionally, you can follow the rinse step depending on whether the composition is formulated as a "stay" or "rinse" product. In order that the present invention may be more readily understood, the following examples are provided only in illustrative sense.

EXAMPLES This example demonstrates the anti-aging benefits of petrosellenic acid. Example 1 n Identification of Procollacene-I and Decorin Disorder in the In Vivo skin following the Topical Treatment of Retinoic Acid for comparative purposes. It is known that collagen, a predominant protein of the structure of the skin, imparts to the same resistance to stress. The decorin is a known proteoglycan that is important for the controlled and correct deposition of the extracellular structure of the skin. It is also known within the art that collagen and decorin levels in the skin are significantly reduced in skin damaged by light or age effects. Many studies have shown that collagen type I levels in the skin decrease with age and / or with increased light effect damage (eg, Lavker, RJ Inv. Derm., (1979), 73, 79- 66; Griffiths et.al. N. Eng, J. Med. (1993) 329, 530-535). In the case of decorin, it has been shown that mRNA expression and proteoglycan expression is greatly reduced in skin damaged by in vitro light effects (Bernstein et al., Lab. Invest. (1995) 72, 662-669 ). The reduction of the levels in these proteins of the skin is associated according to a decrease in the resistance to the tension of the skin, causing wrinkles and sagging. It is well known within the art that retinoic acid is a potent anti-aging agent and induces skin repair of skin damaged by light effects. It has been shown that the prevention of wrinkles and dermal recovery following the topical treatment for the skin with retinoic acid arises through a new deposition of collagen and synthesis in the skin (for example, Griffiths et.al. N. Eng. J. Med. (1993) 329, 530-535). It is widely accepted that strengthening the dermal structure by raising the level of collagen in the skin by using retinoic acid provides recovery benefits against aging / dermal. Procollagen I is a precursor of collagen. The increased production of procollagen I in response to a composite test application is a reference of an increased level of collagen. Two groups of women with identical or nearly identical degrees of sufficient to moderate damage were recruited by effects of light on each outer forearm. They were given 0.05% retinoic acid in a moisturizing base (Retinova®) and also with a matching color-matching moisturizer with similar sensory characteristics (Dermacare® lotion), but without active ingredients, such as a placebo control. Each participant of the two groups applied the Retinova® on one external forearm and the placebo (Dermacare®) on the other external forearm. Group 1 applied the products daily on their external forearms for 14 weeks and Group 2 applied the products on their external forearms for 28 weeks. At the end of the studies, two complete biopsies were taken by 4 mm thick puncture of the treated areas of each forearm. The immunohistochemical analysis of tissue from the biopsy taken from the participants was performed to identify the effect of treatment with retinoic acid on the expression of the extracellular structure components of the skin, decorin and procollagen I, compared with the forearms treated with placebo. The following procedure was followed: MATERIALS An antibody dilution buffer for waxed sections is composed of Tris Tamponated Saline (TBS), 3% bovine wax albumin (BSA), bovine serum albumin, 0.05% Triton X.100 and 0.05% sodium azide. The primary antibodies for the procollagen-1 (amino terminal) were obtained from Chemicon International Inc. (cat # MAB 1912, rat IgG 1) and used in the waxed sections at a dilution of 1: 800; overnight at 4 ° C after the section had been previously treated with trypsin (0.5 mg / ml, 25 minutes, 37 ° C). The primary antibodies for decorin were obtained by Biogenesis (polyclonal rabbit) and used in the waxed sections with a dilution of 1: 800, overnight at 4 ° C. Anti-rat biotinylated secondary antibodies, obtained from DAKO (cat # E0468, rabbit polyclonic), were applied to the waxed sections at a dilution of 1: 400. The biotinylated anti-rabbit secondary antibodies, obtained from Amersham (cat # RPN 1004, donkey polyclonic), were applied to the waxed sections at a dilution of 1: 400. Streptavidin conjugated with alkaline phosphatase, obtained from Zymed (cat # 43-4322), was used at a concentration of 1: 2500. Red Fast Chromogen was obtained from DAKO (cat # K597). Gills # 3 nuclear counterstained haemotoxylin obtained from Sigma (cat # GHS-3), filtered and used without dilution. Trypsin was obtained from Sigma (cat # T-7186) and the slides were mounted with Glycerel from DAKO (cat # C563). METHODS The waxed sections of the biopsy tissue were mounted on silane-coated slides and baked for 18 hours at 55 ° C. The slides were dewaxed through xylan and alcohol and taken to water and transferred to TBS. The DAKO® pen was used to mark the sections. The sections were processed for antigen retrieval with the use of trypsin when necessary, as indicated for each antibody. When antigen retrieval was necessary, the slides were incubated for 25 minutes at 35 ° C with trypsin at 0.5 mg / ml (Sigma Cat # T-7186). Subsequently, the protease was rinsed (2 x 2 minutes) with TBS. Should the following antigen retrieval be necessary, or otherwise directly after labeling the sections, the non-specific antibody binding was not blocked with 5% solutions of a primary serum secondary antibody in TBS / 0.5%. BSA / 0.1% sodium azide as the blocking solution for at least 20 minutes at room temperature in a humid chamber. The excess blocking solution drained outward, but the sections were not allowed to dry. The sections were then incubated with the primary antibody (properly diluted as indicated above) in a humid chamber overnight at 4 ° C. Subsequently, the antibody was drained from the sections, without being allowed to dry. The slides were washed with TBS to remove the unbound primary antibody - a one minute rinse followed by three to five minute washes, and then incubated with the appropriate secondary antibody (properly diluted as indicated above) in a humid chamber during one hour at room temperature. Then, the antibody solution was drained from the slides without allowing the section to dry. The slides were washed with TBS, a one minute rinse followed by 4 x 5 rinses, for the purpose of removing the unbound secondary antibody. For the biotinylated secondary antibody, the sections were incubated, subsequently with streptavidin conjugate for 45 minutes at 37 ° C and then washed in TBS to remove the unbound conjugate of stretavidin. Chromogen was added and the color was developed under observation to avoid excessive staining. The sections were counterstained and mounted. Differences in the expression of procollagen I and decorin between the sites treated with retionoic acid (Retinova®) and placebo (Dermacare®) were determined by visual examinations of sections stained immunohistochemically using microscopic light. This analysis identified a marked disorder in both, procollagen I and decorin, on the skin damaged by effects of light following the topical application of retinoic acid (Retinova®) and the results are set out in the Table 1 that follows. Table 1 Effect of Retinoic Acid Treatment on the expression of procollacene I v decorin on the skin In Vivo The procollagen 1 and decorin components of the extracellular structure are, in this way, clearly identifiable markers of the dermal recovery induced by retinoic acid.

Procedure for Measuring Procollacene-I Synthesis of Decorin in Human Dermal Fibroblasts Preparation of a Conditioned Medium for Fibroblasts Dermal The primary fibroblasts in the human foreskin skin, in step 2 (P2) were cultured in 12-well plates with 1000 cells / cm2 and maintained for 24 hours in a 5% carbon dioxide atmosphere and 4% Oxygen in a Modified Eagles Dulbeccos Medium (DMEM) supplemented with 10% fetal calf serum. After this time the cells were washed with DMEM-free serum and then incubated in a DMEM-free serum for another 60 hours. The individual fibroblast layers were again washed with serum free DMEM. The reagents and vehicle controls were added to the cells in triplicate with a final volume of 0.4 ml / DMEM free of fresh serum per well and incubated for another 24 hours. The conditioned medium of fibroblasts was immediately analyzed or frozen under pressure in liquid nitrogen and stored at -70 ° C for future analysis. Then, the cells were counted and the secant point analysis data were subsequently standardized with the number of the cells.

Drying point test for the protein of Procoláqeno-I v Decorin in a Conditioner with Dermal Fibroblasts. Samples of the conditioned medium with dermal fibroblasts were treated with vehicle (as control) or with test reagents, supplemented with 20mM dithiothreitol (1:10 dilution of 200mM reserve solution), and 0.1% sodium dodecylisulfate ( 1: 100 dilution of 10% reserve solution), mixed well and then incubated at 75 ° C for 2 minutes. A standard for the examination was generated by serial dilution of a fibroblast conditioned medium cleaned from fibroblasts grown at 1000 cells / cm2 in a 175 cm2 flask and maintained in serum free DMEM as described above. The test samples were then applied in triplicate on a premoistened Immobilon-P transfer membrane sheet using a Bio-Rad Bio-Dot 96 well apparatus as described in the manufacturers' guidelines. Approximately 200 μl of medium was applied per well. The medium was allowed to filter through the membrane under the influence of gravity (30 minutes) after which the membrane was washed twice with PBS (200 μl). These PBS rinses were allowed to filter through the membrane under the influence of gravity (2 x 15 minutes). The Bio-Dot appliance was coupled with a vacuum pipe and a third and final rinse of PBS was carried out under the suction effect. The apparatus was decoupled, the membrane removed and cut rapidly as required before being placed in a blocking buffer overnight at 4 ° C. The membranes prepared for the decorin analysis were blocked with 3% (w / v) BSA / 0.1% (v / v) Tween 20 in PBS, while those for the procollagen-1 analysis were blocked with 5% (w / v) dehydrated milk powder without fat / 0.05% Tween 20 in PBS: The next day, the membranes were tested with a 1: 10000 dilution of primary antibodies to human procollagen I (MAB1912, rat monoclonal, Chemicon Int. Inc., Temecula, CA) or for human decorin (rabbit polyclonic, biogenesis) for 2 hours at room temperature. Subsequently, the membranes were washed with TBS / 0.05% Tween 20 (3 x 5 minutes) and then incubated with a 1: 1000 dilution of fragments of 1251-anti-rat or anti-rabbit conjugate F (ab ') 2 ( Amersham) as required for 1 hour at room temperature. After this, the Immobilon strips were again washed with TBS / Tween 20 (3 x 5 minutes) before being allowed to air dry at room temperature. The dried membranes were wrapped in cellophane and exposed to a phosphorus storage screen of Molecular Dynamics for 16-18 hours. After this time, the exposed screen was tracked using a phosphoimage apparatus (Phosphorimager Molecular Dynamics SF) using ImageQuant ™ software, standardized to the number of cells and the effects of the different test reagents in the decorin and procollagen I synthesis, were determined relative to a vehicle control value of 100 arbitrary units.

TESTS Table 2 below indicates the effects of petroselic acid on the synthesis of procollagen I and decorin of human fibroblasts, and the amounts in which they were applied. In order to standardize the results, the effects of the test substance were determined relative to a control value treated with vehicle of 100 arbitrary units. By way of comparison, a study was carried out with retinoic acid to examine its effect on decorin synthesis in human dermal fibroblasts. The concentrations of the reagents used in the studies did not influence the viability of the cell. Table 2 The Effect on the Synthesis of Procoláene I v Decorin with Petrosellenic Acid The results in Table 2 indicate that petrosellenic acid significantly disrupts the synthesis of both procollagen and decorin in human dermal fibroblasts compared to the control. The level of decorin in the skin is associated with an improved condition and appearance of the skin. Increasing the level of decorin in the skin is important for a correct and controlled deposition of collagen in the skin, which is associated with many benefits for the skin such as the prevention of wrinkles and dermal recovery of skin damaged by the effects of light. The comparative study with retinoic acid (1μm) showed a decorin 138+ disorder. 14.0 (p = 0.035, n = 4), determined as relative to a control value treated with vehicle of 100 arbitrary units. Surprisingly, the data also indicate that the extent of the disorder and decorin synthesis in human dermal fibroblasts effected by petroselic acid exceeds that of retinoic acid and the already known reference of active dermal recovery.

Example 2 This example measures the functionality against petrosellenic acid irritations. SDS keratinocyte viability test Methodology Keratinocytes grew in 96-well plates at approximately 80% confluence in a keratinocyte growth medium (KGM) which was replaced with KGM without hydrocortizone for 24-48 hours. Then, the cells were treated with a concentration of sodium dodecyl sulfate (SDS) which would produce approximately 50% cell viability (2μ / ml) in the presence or absence of petrosellenic acid. Then, the cells were dosed with petroselic acid at the concentrations indicated in the following Table 3. The control did not contain any SDS or test compounds. After incubation for 24 hours, the medium was removed and viability was determined by the Neutral Red method. With this method, the cells were incubated for 3 hours in KGM with a content of 25 μg / ml of neutral network after which the medium was removed and the cells were extracted with 1 ml of 1% (v / v) of acetic acid, 50% (v / v) ethanol for 30 minutes. The absorbance of the extract at 562 nm was determined and viability was evaluated with reference to control wells that did not contain SDS or test compounds. The results obtained were summarized in table 3 below: Table 3 All petrosellenic acid (PA) values show significantly increased viability compared to the 2 μg / ml SDS value alone, determined by 1-way ANOVA with multiple Student-Neumann-Kuels comparison, p < 0.05. This methodology has shown that the keratinocyte toxicity of an irritant is related to the irritation effect of the agent in vivo (Lawrence, JN, Starkey, S., Dickson, FM &Benford, DJ.) Use of human keratinocyte cultures and rat to examine the potential of skin irritation Toxicol In Vitro 10, 331-340 (1996)). Thus, it is shown here that the treatment with petrosellenic acid significantly reduces the toxic effects of SDS on keratinocytes and accordingly, that it has a functionality against irritations.

EXAMPLE 3 This example demonstrates that petrosellenic acid can effectively reduce the basal levels of PGE2 (prostaglandin E2) secreted by keratinocytes in vitro.

PEG keratinocyte examination ^ Keratinocytes grew in 96-well plates at approximately 80 confluence in the keratinocyte growth medium (KGM). This was then replaced with KGM without hydrocortizone for 24-48 hours. The cells were then dosed with petroselic acid in the amounts shown in Table 4 below. No petrosellenic acid was added to the control cells. The cells were incubated with (or for control, without) petrosellenic acid for 24 hours. At the end of the incubation, the medium was harvested and examined for basal release of the pro-inflammatory PGE2 by an enzyme binding immunity test using a commercial PGE2 kit (Amersham, Buckinghamshire, England). Then, the cells were tested for viability by the neutral red absorption method described in Example 2 above. The viability of the cells was not found to be adversely effected by the treatment with petroselic acid used at the concentrations tested.

The anti-inflammatory potential of the test compounds was examined by the ability of the components to reduce the baseline levels of PGE2 compared to the control. The statistical significance was determined using a student's t-test. The results obtained are summarized in Table 4 below: Table 4 Statistical analysis using a 1-way ANOVA with a multiple comparison of Student-Neumann-Kuels showed that 0.1 and 1 μM of petrosellenic acid (PA) significantly reduced (p <0.05) the release of PGE2 from unstimulated keratinocytes . PGE2 is a well-known mediator of inflammation in the skin, see Greaves et. to the. "Prostagludus, leukotriemes, phospholipase, plate-driving factor and cytokines: an integrated approach to inflammation of human skin," Arch Dermatol. Res (1988) 280 [Supp]: 533-541. The results indicate that keratinocytes treated with petroselic acid produce less PGE2 of pro-inflammatory prostaglandin, thereby reducing the inflammatory potential of the skin.

EXAMPLE 4 This example measures the effect of petroselic acid on reducing the inflammatory response of dermal fibroblasts. PGE2 Fibroblasts and ICAM Examination Intracellular adhesion molecules (ICAM) and PGE2 production by human skin fibroblasts can be induced by an inflammatory PMA stimulus (myosin acetate). PMA represents an external tensor that induces oxidative stress and inflammatory responses in cells. This model is used for model inflammation in vivo.

The primary fibroblasts of the human foreskin in step 2 (P2) were cultured in 96-well plates at 1000 cells / well and maintained for 24 hours in a 5% carbon dioxide atmosphere in a Modified Dulbeccos Eagles Medium (DMEM) supplemented with 10% fetal calf serum. heifer. Petrosellenic acid was added to the fresh cell medium (DMEM, supplemented with 10% cow's milk fetus serum) in dimethylsulfoxide (DMSO, final concentration 1%) by tricipylate and incubated for another 24 hours. The myrmetal acetate (PMA) (Sigma) was added to the medium and the cells were incubated for another 24 hours. The control did not contain any test compound or PMA. The fibroblasts / medium were analyzed as described below, immediately or were snap frozen in liquid nitrogen and stored at -70 ° C for future analysis. Then, the cells were counted and the secant point analysis data were subsequently standardized for the cell number. Prostaglandin E2 (PGE2) Test: The 50 μl volume of the culture medium was taken for a PGE2 test after gently mixing the culture plate. The levels of PGE2 in the medium were determined with a PGE2 Biotrak immunoexamination kit (Amersham, UK). The test was based on the competition between the PGE2 labeled in the sample and a fixed amount of horseradish peroxidase labeled PGE2 for a limited amount of specific PGE2 antibody. The concentrations of the sample not labeled PGE2 are determined according to a standard curve that was obtained at the same time. ICAM-1 test: The medium was discarded and the cells were washed with Dulbecco PBS. For the washed cells, 150 μl 0.1% Triton X-100 (Sigma) was added during 3 minutes to extract ICAM from the cell membrane. The extracts were transferred to Eppendoff centrifuge tubes and centrifuged at 1000 g for 2 minutes to remove dirt from the cell. A volume of 100 μl of supernatant was used for the ICAM examination. Soluble ICAM-1 was examined with an immunoenzymometric kit available for sale (R & amp; amp;; D Systems). The ICAM-1 concentrations in the samples were determined based on a standard curve that ran in parallel. The results obtained from the PGE2 and ICAm exams are summarized in Table 5 below. Table 5 Effects of petroselic acid on the production of ICAM and PGE ^ PMA-Induced in human skin fibroblasts. * p < 0.001 compared with those of cells treated with PMA.

The above results show that the cells tested with inflammatory stimulants such as PMA (myristyl myristate acetate) causes an increase in the inflammatory response as measured by the production of prostaglandin (PGE2). Petrosellenic acid even at 0.1μm levels. dramatically reduces the inflammatory response as measured by the production of PGE2. In this way, the results show that petrosellenic acid has a good activity against inflammation. The above results also demonstrate that cells tested with PMA cause an increase in ICAM production. Petrosellenic acid decreases the production of intramolecular adhesion molecules (ICAM), which is another marker of inflammation. Thus, these results also show that petrosellenic acid has good action against inflammation.

EXAMPLE 5 METHODOLOGY OF THE EXAMINATION FOR LIGHTING THE SKIN Maintenance of the cells Some melanoma cells of mouse B16-F1 (American Type Culture Collection, Maryland, USA) were maintained in 75 cm2 culture flasks in an RPMI 1640 medium (Flow-ICN, cat No. 12-60-54) supplemented with L-glutamine (4 mM) and 10% Bovine fetus serum (FBS) at 37 ° C in saturated water, 5% CO2 in atmospheric air. The cells were passed twice a week. Pigmentation test. Subconfluent B16 cells were grown in microtiter plates at a density of 5000 cells / well and cultured overnight in DMEM (Life Technologies, NY) containing 10% bovine fetal serum and 1% penicillin / streptomycin. phenol red at 37 ° C under 5% CO2. After 24 hours, the medium was replaced with a fresh medium with a content of test materials or with vehicle controls. The cells were incubated for 72 hours, the lapse of which allowed the melanin to become visible in the control wells. Then, the medium with melanin content from each well was transferred to a clean 96-well plate and quantified by reading the absorbance at 530 nm using a microplate spectrophotometer (Dynatech MR5000) and correcting for baseline absorption of the fresh medium. Because the corrected absorption is proportional to the concentration of melanin, the percentage of pigmentation for a test substance to lighten the skin can be calculated as follows:% pigmentation = (OD530 test / OD530 ref) x 100%; wherein, the OD530 test and the OD530 ref indicate the average corrected absorption of the medium from the wells with a test substance and that from the medium of the wells without the test substance. The percentage of inhibition caused by the test substance is then 100% pigmentation. Feasibility study of the cell. Melanin production can be reduced by the inhibition of melanogenesis but it can also be affected by cytotoxicity or cell proliferation. To test if this occurred, the viability of the cell was tested by an absorption of neutral red dye. Neutral red is a water-soluble dye that passes through the intact membrane of the plasma and is concentrated in lysosomes in intact cells. The total absorption of the neutral red dye is proportional to the number of viable cells in culture. Immediately after the removal of the medium for the melanin analysis of the microtiter wells, 200 μl of preheated neutral red dye (eg Sigma, RU, Cat. Nr 2889) in a medium of 25 μg / ml was applied to the cells and they were incubated during 3 hours for the maintenance of the cells. The dye that had not been absorbed by the cells was removed by inverting the plate and emptying it on absorbent paper. The cells were washed with 200 μl PBS, which was removed again. They were added 100 μl of solvent (50% H2O, 49% ethanol, 1% acetic acid). After 20 minutes at room temperature, each plate was stirred for 5 seconds on a microtiter plate shaker. Absorption was measured as explained above.

Tests Table 6 below indicates the skin lightening substances evaluated and the amount in which they are applied. The percentage of inhibition of melanin production caused by the test substances as described above is also reflected in the table. The values lower than a 100% control of melanin indicate the inhibition of melanogenesis. In this way, the results in Table 3 show that PA inhibits melanin production. In the studies, the test substance was diluted with DMEM in the amounts shown in Table 5 below.

Test t student ** p < 0.01 p < 0.05 (n = 4) Example 6 The following formula describes an oil-in-water cream suitable for the methods and uses in accordance with the present invention. The percentages indicated are by weight of the composition.

* Brij 56 is cetyl alcohol POE (10) Alfol 16RD is cetyl alcohol Example 7 The following formulation describes an emulsion cream according to the present invention.

Both topical compositions of Example 6 and 7 provide an effective cosmetic treatment to lighten the skin color and / or to improve the appearance of wrinkled, aged skin, damaged by light and / or irritated skin, when applied to the skin which has deteriorated due to age and time that has been exposed to light or when applied to young skin to help prevent or slow down such deterioration changes. The compositions can be processed in a conventional manner.

Claims (2)

1. CLAIMS 1. A cosmetic method for providing at least one skin care benefit selected from: treatment / prevention of wrinkles, sagging, aged skin and / or damaged by effects of light; high deposition of collagen in the skin, high production of decorin in the skin, improvement in tissue recovery; rinsing of the skin; relieve irritations, red and / or sensitive skin; improve the texture of the skin; softness and / or firmness; The method comprises applying to the skin a topical composition comprising petrosellenic acid and / or derivatives thereof.
2. 2. The use of petrosellenic acid and / or derivatives thereof for the preparation of a topical composition to provide at least one benefit to the selected skin of: treatment / prevention of wrinkles, sagging, aged skin and / or damage by effects of the light; high deposition of collagen in the skin, high production of decorin in the skin, improvement in tissue recovery; rinsing of the skin; relieve irritations, red and / or sensitive skin; improve the texture of the skin; softness and / or firmness.