FR2921267A1 - Use of an activator of perilipin in a slimming cosmetic composition, as slimming agent, anti-cellulite agent, anti-orange skin agent and/or lipolysis stimulator, and for the preparation of dermatological composition for slimming - Google Patents

Abstract

Use of an activator of perilipin in a slimming cosmetic composition, as a slimming agent, anti-cellulite, antiorange skin and/or lipolysis stimulator, is claimed. ACTIVITY : None given. MECHANISM OF ACTION : Perilipin activator.

Description

Slimming cosmetic composition

The present invention relates to the field of cosmetics and more particularly the field of slimming. The present invention relates more particularly to the use of a perilipine activator in a slimming cosmetic composition.

The adipose tissue (or hypodermis) is attached to the lower part of the dermis by expansions of collagen fibers. Its thickness is variable, thin on the forehead, it develops largely on particular areas (thighs, abdomen). The anatomical location of adipose tissue is a true secondary sex trait. In men, it is predominant above the waist, in the abdomen and shoulders, whereas in women, it is concentrated below the belt, in the lower abdomen and in the abdomen. level of the hips, buttocks and thighs. This sex-related location is strongly emphasized in the case of obesity, of which two forms are distinguished, the android form and the gynoid form.

Adipose tissue consists of: - specific cells, adipocytes that have the ability to store fat (lipogenesis) but also to mobilize (lipolysis), - an extracellular matrix, - a dense capillary network. The development of adipose tissue is characterized by a modification of all the components of the hypodermic tissue. These disturbances also have repercussions on the more superficial layers. At the level of the dermis, an increase in thickness is observed, probably due to fluid engorgement (water retention) and infiltration of adipose lobules deforming the dermis / hypodermis junction; on the surface the skin takes a padded appearance is the famous orange peel.

The development of adipose tissue involves different mechanisms: - the reorganization of the extracellular matrix, - the differentiation of pre-adipocytes into adipocytes, - the increase of the volume of adipocytes.

Lipolysis corresponds to the catabolism of the triglycerides stored in the adipocytes, that is to say their degradation to non-esterified fatty acids and to glycerol. The molecules thus released into the blood stream can be used by other tissues as an energetic substrate (fatty acids) or as neoglucogenic precursors (glycerol). The mobilization of fatty acids from the triglycerides stored in adipose tissue requires the activation of a lipolytic enzyme, the hormone-sensitive lipase (LHS). Hormone-sensitive lipase hydrolyzes triglycerides to diglycerides and diglycerides to monoglycerides, it is the enzyme limiting lipolysis. Monoglyceride lipase (LMG) then degrades monoglycerides to non-esterified fatty acids and glycerol. The activity of LHS is tightly controlled by reversible phosphorylation by a cAMP-dependent protein kinase 2 (PKA) or cGMP (PKG). Moreover, the phosphorylation of the LHS allows a redistribution of the lipase of the cytosolic compartment towards the lipid vacuole allowing it to act on the triglycerides. Perilipines are proteins mainly expressed on the surface of the lipid vacuole and probably intervene in its stabilization. The Applicant has demonstrated the action of Scabiosa arvensis on the activity of perilipin. Indeed an extract of Scabiosa arvensis makes it possible to phosphorylate perilipine. Activation of perilipine allows the LHS to undergo functional rearrangement and colocalize on the lipid vacuole alongside perilipin. It can thus enter the vacuole to hydrolyze triglycerides. As a result, an extract of Scabiosa arvensis acts as a slimming active ingredient when applied to the skin by means of a cosmetic composition.

The application therefore relates to the use of a perilipine activator in a slimming cosmetic composition. The application also relates to the use of a perilipine activator in a cosmetic composition as slimming agent and / or anti cellulite and / or anti orange peel and / or stimulator of lipolysis.

The activity of a perilipine activator is evaluated by the test described below: - culturing of pre-adipocytes 30 - incubation of the cells with the active - detection by immunofluorescence of the protein rearrangement of perilipin and the LHS as illustrated in Example I.

This activator may be in the form of a plant extract, more particularly a plant extract of the family Dipsacaceae. Preferably, it is an extract of Scabiosa arvensis.

Scabiosa arvensis, commonly known as scabiosa, is a perennial herb with upright growth of 30 cm to 1.30 m tall. The leaves arranged in a rosette are simple, lanceolate, more or less cut, opposite and hairy as the stem. The Parma-colored flowers are grouped into a capitula 15 to 40 mm in diameter. Originally from Eurasia, common in Europe, the scabious colonizes meadows and woods up to 1900m breeding by dispersing its seeds. In the invention, the aerial part of the plant is used. The extract of Scabiosa arvensis is obtained by the implementation of a process comprising at least one step of solubilizing Scabiosa arvensis and at least one enzymatic hydrolysis step. An aqueous-alcoholic extract of Scabiosa arvensis is obtained, preferentially a hydro-glycerine extract of the plant. It is a clear, yellow, low-odor liquid with the following characteristics - pH = 4.6 - solids = 512.7 g / l 4 - total sugars 9.3 g / l - total polyphenols = 0, 17 g / 1

It is possible to obtain an extract of Scabiosa arvensis that can be used according to the invention, for example from the company SILAB, marketed under the name REDUCELL 2.

The composition according to the invention contains from about 0.01 to 10% by weight, and preferably from 0.01 to 5% by weight of an extract of Scabiosa arvensis.

According to the invention, the extract of Scabiosa arvensis can be used in combination with one or more other slimming active agents. More particularly, the extract of Scabiosa arvensis according to the invention can be combined in a cosmetic composition with a compound chosen from caffeine, a geranium extract, an extract of Baccharis genistelloides, a small holly extract, a brown extract of India, an extract of Ginkgo biloba or a red vine extract. In particular, the extract of Scabiosa arvensis can be combined in a cosmetic composition with caffeine. Similarly, the extract of Scabiosa arvensis may be combined in a cosmetic composition with a geranium extract as described in patent FR 2804319. Finally, the extract of Scabiosa arvensis may be combined in a cosmetic composition with an extract of Baccharis genistelloides As described in FR 2867074 The cosmetic composition according to the present invention may contain one or more other components known to those skilled in the art, such as formulating agents or additives of known and conventional use in cosmetic compositions. By way of example and without limitation, such formulating agents and additives may be hydrophilic or lipophilic gelling agents, softeners, colorants, solubilizing agents, texturizing agents, perfumes, fillers, film-forming active agents , preservatives, surfactants, emulsifiers, oils, glycols, vitamins, sunscreens, .... Thanks to his knowledge of cosmetics, the skilled person will know which formulating agents to add to the cosmetic composition according to the invention and in what quantities according to the desired properties.

In addition, the cosmetic composition according to the present invention may be in any form known to those skilled in the field of cosmetics without any other particular pharmaceutical restriction than that for application to the skin. Thus, the cosmetic composition according to the invention may have the form of an aqueous solution, or an alcoholic suspension or an oily suspension or a solution or dispersion of the lotion or serum type, of a consistency emulsion. liquid or semi-liquid milk type, obtained by dispersion of a fatty phase in an aqueous phase (oil in water emulsion: O / W) or conversely (water in oil: W / O), or an emulsion of the type O / W or W / O cream or gel, lotion, mask. The 6 cosmetic formulations according to the invention can also be envisaged in the form of a foam or in the form of aerosol compositions also comprising a propellant under pressure.

The present invention also relates to a cosmetic slimming treatment method, comprising the application to the skin of a cosmetic composition comprising a perilipine activator. The present invention also relates to the use of a perilipine activator for the preparation of a dermatological composition for body thinning.

The following examples concern, on the one hand, the evaluation of the activation of perilipine by an extract of Scabiosa arvensis, and, on the other hand, the compositions that are the subject of the present invention. The examples refer to the following figures in which: FIGS. 1a to 3c represent the visualization of the colocalization of perilipin and LHS for basal cells. Figure la represents the visualization of perilipin. Figure 1b shows the visualization of the LHS. Figure 1 represents the visualization of the superposition of the two LHS and perilipine markers as well as the result of the image analysis by the LUCIA ° software. FIGS. 2a to 2c show the visualization of the colocalization of perilipine and of the LHS for cells treated with isoproterenol 100e. Figure 2a shows the visualization of perilipin. Figure 2b shows the visualization of the LHS. Figure 2c shows the visualization of the superposition of the two LHS and perilipine markers as well as the result of the image analysis by the LUCIA ° software. Figure 2d shows the diagram of the percentage of colocalization of the two markers LSH and perilipin.

FIGS. 3a to 3c show the visualization of the colocalization of perilipine and LHS for cells treated with 0.11% of an extract of Scabiosa arvensis. Figure 3a shows the visualization of perilipin. Figure 3b shows the visualization of the LHS. Figure 3c shows the visualization of the superposition of the two LHS and perilipine markers as well as the result of the image analysis by the LUCIA ° software. Figure 3d shows the diagram of the percentage of colocalization of the two markers LSH and perilipin. FIGS. 4a to 4c show the visualization of the colocalization of perilipin and LHS for cells treated with 0.33% of an extract of Scabiosa arvensis. Figure 4a shows the visualization of perilipin. Figure 4b shows the visualization of the LHS. Figure 4c shows the visualization of the superposition of the two LHS and perilipine markers as well as the result of the image analysis by the LUCIA ° software. Figure 4d shows the diagram of the percentage of colocalization of the two markers LSH and perilipin. 8 I. EVALUATION OF PERILIPIN ACTIVATION BY SCABIOSA ARVENSIS A.MATERIAL EXTRACT AND METHOD 1. Culture of 3T3-L1 pre-adipocytes a) Culture media: The culture medium used is a complete nutrient medium such as modified essential medium Dulbeco (DMEM). Due to the intensity of carbohydrate metabolism in these cells, we supplement the medium with glucose at a final concentration of 4.5 g / l (Gibco). Standard culture conditions for preadipocytes require supplementation of the medium with 10% fetal calf serum (Cambrez). Under these conditions, attachment, cell growth and adipocyte differentiation are satisfactory. b) Evolution of cultures, proliferative rhythms and differentiation: The pre-adipocytes, for our experiment, are seeded in 6-well plates with glass coverslip at the rate of 20,000 cells per dish. These multiply until confluence, a stage reached 6 days after seeding. After the confluence, the cells stop multiplying and we attend the implementation of a program of differentiation by the addition to the nutritive medium, of insulin (Sigma) (1,7} mol / L), of IBMX (Sigma) (0.5 mmol / L) and dexamethasone (Sigma) (1 pmol / L). The cells round up and begin to accumulate triglycerides in the form of droplets that coalesce with each other. 2 - Procedure of the study The cells are rinsed and incubated for 24 hours in a DMEM medium containing 2 mM L-glutamine (Gibco) and 0.1% BSA (bovine serum albumin) (PAA), a nutrient medium used to the study.

The cells are rinsed and incubated for 3 hours as follows. . Middle only (control). Isoproterenol 100 gM (Sigma) (positive reference chosen for its stimulating action of the non-specific adrenergic-PKA pathway). . Scabiosa arvensis 0.11% * in medium. Scabiosa arvensis 0.33% * in medium (final volume 2 ml) (2 wells per condition) * concentration revealed non-cytotoxic for MTT cells (Mosmann, J Immunol Meth 1983 65: 55-63). Each cell mat is then rinsed, fixed with methanol (-20 ° C) before revealing perilipine and the hormone-sensitive lipase by immunofluorescence. 3 - Immunofluorescence: Immunolocalization of LHS and Perilipin (Plin) by Double Labeling Reactions The double labeling reactions are very simple to carry out when antibodies are available.

obtained from different animal species and secondary antibodies specific for these species, labeled with fluorescein isothiocyanate (FITC) or rhodamine isothiocyanate (TRITC).

The cells fixed in methanol at -200C for 4 minutes and then in 3.7% formaldehyde for 10 minutes are permeabilized in a solution of 0.1% Triton X100 in PBS for 10 minutes at room temperature. Nonspecific sites are saturated with a solution of 2% BSA in PBS (phosphate buffered saline) for 10 minutes at room temperature. The cells are incubated with the anti-perilipine antibody (Acris antibody, dilution 1: 100) (source: guinea pig) and then with the secondary anti-guinea pig antibody coupled to TRITC (tetramethylrhodamine isothiocyanate Abcys, dilution 1:75). In a second step, the cells are incubated with the 2nd anti-LHS antibody (Cayman chemical, dilution 1: 100) (source: rabbit) and then with the FITC-coupled anti-rabbit secondary antibody (fluorescein isothiocyanate Zymed, dilution 1 : 100). The incubation time between each antibody is 1 hour. Between each incubation of antibodies, 3 rinses with PBS of 3 minutes each are necessary.

The slides are mounted on a glass slide and then observed under a fluorescence microscope (Nikon Eclipse 50i). Photographic snapshots are analyzed by the Lucia image analysis software.

B.RESULTS 11

Perilipin is detected by a guinea pig antibody followed by a secondary anti-guinea pig antibody coupled to a red fluorochrome (TRITC). It appears in red. LHS is detected by a rabbit antibody followed by an anti-rabbit secondary antibody coupled to a green fluorochrome (FITC). It appears in green. The superposition of the two perilipin and LHS markings identifies the colocalization area appearing in yellow. The colocalization of the two markers is a sign of a rearrangement of perilipine and LHS. Isoproterenol is a structural analogue of catecholamines where the amine function is substituted by an isopropyl group. The greater congestion of the amine function, compared to that of norepinephrine and even that of adrenaline, makes the substance lose its affinity for the n-adrenergic receptors, but retains its ability to be recognized by the receptors. R-adrenergic and to stimulate them. Isoproterenol stimulating the p-adrenergic-PKA pathway allows concomitant phosphorylation of perilipine and LHS. Under these experimental conditions, there is a clear marking of perilipin around each droplet in the case of adipocytes treated with isoproterenol compared to the control, sign of the rearrangement of perilipine phosphorylated so that the LHS can be rearranged. Indeed, the LHS is positioned around the droplet, co-locating with its coactivator, perilipin. Microscopic observation shows that perilipine and LHS are largely co-localized to the surface of the lipid vacuole when the adipocytes are treated with isoproterenol. The positive reference validates the experiment. The extract of Scabiosa arvensis applied to adipocytes has the same effects as isoproterenol. We note an intense labeling of perilipine and LHS at the periphery of lipid droplets. Observation by microscopy shows that perilipine and LHS are very largely localized at the periphery of the lipid droplets. The areas of interest are predominantly yellow. Scabiosa arvensis induced the phosphorylation of perilipines, thus promoting access of activated LHS to its lipid substrates.

B. CONCLUSION Scabiosa arvensis controls, via the activation of adipocyte plasma membrane receptors, the LHS and its corresponding coactivator, perilipin, proteins which are themselves essential for the activation of lipolysis. II.EXAMPLES SLIMMING LOTION o Glycol 30.00 Preservative 0.50 Caffeine 1.00 Geranium extract 1.00 13 Baccharis genistelloides extract 1.00 Scabiosa arvensis extract 1.00 Demineralized water QSP 100 SLIMMING GEL ° Carbomer 0.39 Alcohol 96.2 ° 41.50 Caffeine 1.98 Geranium extract 1.50 Peruvian liana extract 3.00 Agrimony extract 0.75 Baccharis genistelloides extract 3.00 Neutralizing base 0.18 Scabiosa arvensis extract 1 , 00 Fragrance ... 0.50 Demineralized water QSP 100 SLIMMING EMULSION ° 0 25 Carbomer 0,50 Acrylic polymer 5,00 Caffeine 2,00 Baccharis genistelloides extract 1,00 Scabiosa arvensis extract 1, 00 30 Agrimony extract 0.25 Peruvian liana extract 1.00 Geranium extract 0.50 Silicone 1.50 10 15 20 14 10 Ester g-as 1.50 Silicone volatile 6.00 HeliogeL 0.50 Soda 0.15 Alcohol 96.2 ° 15,00 Menthol 0,35 Scent 0,50 Demineralized water QSP 100 15

Claims (8)

1. Use of a perilipine activator in a slimming cosmetic composition. 2. Use of a perilipine activator in a cosmetic composition as a slimming agent and / or anti cellulite and / or anti orange peel and / or stimulator of lipolysis. 10 3. Use according to claim 1 or 2 characterized in that this activator is in the form of a plant extract. 15 4. Use according to claim 3 characterized in that this activator is in the form of a plant extract of the family Dipsacaceae. 5. Use according to claims 3 or 4 characterized in that this activator is an extract of Scabiosa arvensis. 6. Use according to claim 5 characterized in that the Scabiosa arvensis extract is an extract of the aerial parts of the plant. 7. Use according to claims 5 or 6 characterized in that the extract of Scabiosa arvensis is a hydro-alcoholic extract of the plant. 8. Use according to claims 5 to 7 characterized in that the extract of Scabiosa arvensis is a hydro-glycerine extract of the plant. 16. Use according to claims 5 to 8, characterized in that the composition contains from about 0.01 to 10% by weight, and preferably from 0.01 to 5% by weight of an extract of Scabiosa arvensis. . 10. Use according to any one of the preceding claims, characterized in that the composition further contains a compound selected from caffeine, a geranium extract, an extract of Bocchoris genistelloides, a small holly extract, a brown extract of India, an extract of Ginkgo biloba or a red vine extract. 11. Cosmetic slimming treatment process, comprising the application to the skin of a cosmetic composition comprising a perilipine activator. 12. Use of a perilipine activator for the preparation of a dermatological composition for body thinning. 30 17