FR2961692A1 - Cosmetic use of extract of edible mushroom stem including extract of mushroom or bolete, for treating skin aging, by applying topical composition containing extract and vehicle, preferably for providing antiglycation effect - Google Patents

Abstract

Cosmetic use of an extract of edible mushroom stem including an extract of mushroom or bolete, for treating skin aging by applying a topical composition containing at least an extract and vehicle, preferably for providing antiradical- and antiglycation-effect and/or stimulating the synthesis of filaggrin, is claimed. An independent claim is included for a cosmetic composition for topical application comprising vehicle and the extract of at least one edible mushroom stem having dry material content of 0.1-100 g/l, preferably 1-50 g/l, pH of 3-8, preferably 4-6, protein content of 0.0005-10 g/l, preferably 0.001-1 g/l, total sugar rate of 0.01-100 g/l, preferably 0.05-50 g/l and total phenol rate of 0.001-50 g/l, preferably 0.005-10 g/l and comprising an oligosaccharide fraction (50-95 (preferably 70-90) wt.%) of the total carbohydrate fraction, consisting of glucose (65-99 (preferably 75-98) mole%) of the total carbohydrate fraction and optionally including traces of rhamnose and fucose; a polysaccharide fraction (2-50 (preferably 5-30) wt.%) of the total carbohydrate fraction, comprising a minimum 50%, preferably 60% glucose and 15-35%, preferably 20-30% of mannose and optionally including traces of xylose and galactose, and optionally including traces of xylose and galactose; and a polysaccharide fraction mainly composed of glucan having a molecular weight of 500-65000 Da. ACTIVITY : Dermatological. MECHANISM OF ACTION : Filaggrin synthesis stimulator.

Description

The present invention relates to the cosmetic use of an extract derived from edible pedunculated mushrooms, such as porcini mushrooms, edible mushrooms of the family of boletus, as well as cosmetic compositions comprising them and a cosmetic process using them.

Many cosmetics have for the moment been proposed for the care of the skin, and in particular its anti-aging treatment. Several mechanisms involved in this area have been highlighted. Human skin consists of three layers: - the superficial layer, the epidermis, consisting mainly of keratinocytes and corneocytes. - The dermis, consisting mainly of fibroblasts, is based on the hypodermis which joins the subcutaneous anatomical structures. Rich in connective fibers mainly collagen and elastin, the dermis confers the strength and elasticity of the skin. - the hypodermis, a richly vascularized loose connective tissue which, depending on the nutritional conditions and the regions of the skin, contains more or less adipose tissue. The phenomenon of skin aging can be accelerated by repeated exposure to solar radiation or atmospheric pollutants. These factors lead to the formation of free radicals. A free radical is an atom or molecule that has won or lost an electron. It is this odd number of electrons that makes the molecule unstable. This one keeps catching or giving up an electron to another molecule of its entourage, thus spreading the phenomenon. In the body, this reaction is called oxidative stress. The radicals produced are the superoxide anion, the hydrogen peroxide and the hydroxyl radical. These reactive oxygen species can alter cell membranes, DNA, proteins and lipids. The interest of an active radical scavenger can be evaluated by the 1,1-diphenyl-2-picrylhydrazyl (DDPH) test.

In addition, skin aging is promoted by glycation reactions. Glycation is a so-called non-enzymatic Maillard reaction, which corresponds to the binding of a reducing ose or an aldehyde to the amino residue of a protein to form a Schiff base. This can cause, after a molecular rearrangement of Amadori, an intermolecular bridging. Glycation makes the proteins more resistant to proteolysis thus preventing their renewal. This phenomenon occurs particularly at the level of collagen fibers which lose their solubility and become difficult to degrade. Thus, a stiffening of the tissues causes a loss of tone of the skin.

The ability to inhibit glycation products reveals for an active ingredient its interest in the elasticity of the skin.

Finally, the barrier function of the skin is ensured by the stratum corneum, superficial layer of the epidermis. It allows protection of the underlying layers against external aggressions such as exposure to solar radiation, chemical and atmospheric pollutants. On the surface of the stratum corneum are the corneocytes, resulting from a differentiation of keratinocytes. These corneocytes contain filaggrin, a protein that plays an important role in the aggregation of keratin filaments and ensures the differentiation of keratinocytes into corneocytes. Upon degradation, filaggrin releases amino acids and their derivatives that will be metabolized to form the NMF (Natural Moisturizing Factor) components. NMFs are responsible for maintaining the hydration of the stratum corneum. A decrease in the synthesis of filaggrin following cutaneous aging can thus result in a loss of water and consequently a drying of the skin. The ability of an active ingredient to increase the expression of filaggrin reveals its interest in enhancing the barrier function of the skin.

It is known that certain natural extracts have antioxidant properties. Thus, extracts of Boletus edulis have already been described (Tsai et al., [11]), in particular for their anti-oxidant properties, in the food field. However, the incorporation of these extracts into cosmetic compositions is neither described nor suggested in this document.

Moreover, the moisturizing effect of various extracts including a boletus extract is mentioned in JP-2004 224702. However, their use in the anti-aging treatment of the skin, especially for anti-radical, anti-glycation or stimulation of the synthesis of filaggrin are not described or suggested.

There is still a need for active ingredients at least for the activities described above, namely stimulation of filaggrin synthesis, free radical uptake and / or inhibition of glycation phenomena. There remains also a need for cosmetic compositions and active or active ingredients, of natural origin, accessible, easy to isolate and whose preparation is reproducible, easy to formulate and cosmetically acceptable and which also have at least some of the cosmetic properties interesting above. The Applicant has now shown that edible pedunculated mushroom extracts respond to the above problems. In particular, it has been shown that mushroom extracts not only have the property of capturing free radicals, thus limiting cell aging, but they also inhibit glycation phenomena and induce stimulation of filaggrin synthesis. The extracts that can be used according to the invention thus meet the needs mentioned above.

This is why the present invention relates to the cosmetic use of pedunculated edible mushroom extracts. These extracts are used for the anti-aging treatment of the skin by application of a topical composition comprising at least one such extract and a cosmetically acceptable vehicle, in particular for anti-radical, anti-glycation and / or stimulation purposes. synthesis of filaggrin. In addition, the present invention relates to cosmetic compositions comprising at least one pedunculated edible mushroom extract and a cosmetically acceptable carrier. It also relates to a cosmetic process comprising the application of compositions according to the invention.

Among the pedunculated edible fungi, it is possible to use, according to the invention in particular, the boletes, and among these, Boletus edulis, Boletus erythropus, Boletus badius, Boletus luridus, Suillus bovinus, Sui / lus granulatus, Leccinum scabrum, Boletus aereus, Boletus pinicola and Boletus aestivalis and their mixtures. As will appear in the examples, an active agent that can be used according to the invention was obtained by an aqueous extraction of the water-soluble molecules of a mixture comprising Boletus edulis, for the most part, and Boletus aereus, Boletus pinicola and Boletus aestivalis, ie say mushrooms commonly called "cep".

The extraction of the molecules of interest can be carried out by any method compatible with the preservation of their properties, from fresh or dry material. In a particular embodiment, an extract usable according to the invention, hereinafter "extract", can be obtained from the fungus dry matter incorporated at a suitable concentration in a suitable extraction solvent, for example in the following manner: water, by refluxing and separating by any known means, for example by centrifugation. It is also possible to provide hydroalcoholic extracts with alcohols such as alkanols, especially ethanol. In a particular embodiment of the invention, the extract, which may be aqueous, meets at least one of the following criteria: a) dry matter content, determined by lyophilization, of between 0.1 and 100 g / L, preferably between 1 and 50; b) pH of between 3 and 8, preferably between 4 and 6; c) protein level between 0.0005 and 10 g / L, preferably 0.001 and 1 g / L; d) total sugar levels between 0.01 and 100g / L, preferably between 0.05 and 50g / L; e) total phenol content between 0.001 and 50 g / l, preferably 0.005 and 10 g / l; f) an oligosaccharide fraction representing between 50 and 95% by weight of the total carbohydrate fraction, preferably composed of 65 to 99 mol% of glucose, preferably between 70 and 90% by weight of the total carbohydrate fraction, preferably composed of 75 to 98% by weight; mol% of glucose and optionally comprising traces of rhamnose and fucose; g) a polysaccharide fraction representing between 2 and 50% by weight of the total carbohydrate fraction, preferably comprising at least 50% glucose and 15 to 35% mannose and optionally comprising traces of xylose and galactose, preferably representing between 5 and 30% by weight of the total carbohydrate fraction, preferably comprising at least 60% glucose and 20 to 30% mannose and optionally comprising traces of xylose and galactose; h) a polysaccharide fraction consisting essentially of glucans with a molecular mass of between 500 and 65,000 Da.

In a more preferred embodiment of the invention, the extract meets at least one of the following criteria: a) dry matter content, determined by lyophilization, of between 15 and 25 g / l, preferably 19, 34 ± 0.20 g / L; b) pH of between 3 and 8, preferably between 4 and 6; c) protein level of 0.02 ± 0.01 g / L; d) total sugar levels ranging from 1 to 25 g / L, preferably 7.99 ± 1 g / L; e) total phenol content between 0.1 and 2.5 g / L, preferably 0.85 ± 0.05 g / L; f) an oligosaccharide fraction isolated by steric exclusion chromatography on BIO-RAD CL4B gel, representing between 70 and 90% by weight of the total carbohydrate fraction. It is composed of 75 to 98 mol% of glucose and contains some traces of rhamnose and fucose, composition determined by gas chromatography equipped with a capillary column (0.32mm * 50m) CP-SIL-5CB and a detector flame ionization (FID); g) a polysaccharide fraction representing between 5 and 30% by weight of the total carbohydrate fraction, comprising at least 60% of glucose and 20 to 30% of mannose, and possibly traces of xylose and galactose; h) a polysaccharide fraction consisting essentially of glucans with a molecular mass of between 1000 and 50 000 Da.

Thus, the present invention relates to the use of at least one pedunculated edible mushroom extract to stimulate the synthesis of filaggrin, thereby promoting the hydration of the skin and / or reducing the glycation phenomena responsible for the loss of elasticity of the skin. skin and reduce the phenomena of oxidative stress responsible for premature aging of the skin. The extracts can be used for anti-glycation, filaggrin stimulation, or anti-free radical purposes. In particular, it has been found that the extracts which can be used according to the invention have in particular significantly increased the expression of filaggrin in a concentration-dependent manner. They therefore have a pro-differentiating effect, and thus make it possible to reinforce the epidermal barrier function. Other advantages, properties and characteristics of the extracts of the invention will appear in the examples below and in the accompanying figures. FIG. 1 illustrates the anti-radical activity of an extract that can be used according to the invention compared with a control. FIG. 2 illustrates the anti-glycation activity of an extract that can be used according to the invention in comparison with a control.

For example, the mushroom extracts can be obtained from dry matter introduced in a proportion of 0.2 to 20% by weight and more particularly of 3.33 ± 1% by weight in water. This extraction can be carried out at reflux at 70-90 ° C. and more particularly at 80 ° C. for 15 to 60 min and more particularly for 30 minutes. The extract is recovered by centrifugation at 3500 rpm for 10 minutes.

The following characteristics can be highlighted especially using techniques as indicated below. The extract has: a solids content of between 1 and 50 g / l. This rate is determined by lyophilization of the material. - A pH between 3 and 8 and more particularly between 4 and 6. The pH is determined by a potentiometric method. A protein level of between 0.001 and 1 g / l. This rate is determined by a colorimetric assay according to the Bradford method [1]. - A total sugar level between 0.05 and 50g / L. This rate is determined by a colorimetric assay according to the method of Dubois [2] and Blumenkrantz [3]. A total phenol content of between 0.005 and 10 g / l. This rate is determined by a colorimetric assay according to the method of Singleton and Rossi [4]. An oligosaccharide fraction isolated by steric exclusion chromatography on BIO-RAD CL4B gel, representing between 70 and 90% by weight of the total carbohydrate fraction. It is composed of 75 to 98 mol% of glucose and contains some traces of rhamnose and fucose, composition determined by gas chromatography equipped with a capillary column (0.32mm * 50m) CP-SIL-5CB and a detector flame ionization (FID). These results were obtained by trimethylsilylation of monosaccharides according to the Kamerling method [5]. A polysaccharide fraction isolated by steric exclusion chromatography on a BIO-RAD CL4B gel representing between 5 and 30% by weight of the total carbohydrate fraction. It is composed of at least 60% glucose and 20 to 30% mannose, and includes some traces of xylose and galactose, composition determined by gas chromatography. This fraction consists essentially of glucans whose molecular mass is between 1000 and 50000 Da [6]. The mixture extract or extracts are used according to the invention in a composition containing a cosmetically or dermatologically acceptable medium, that is to say a medium that is compatible with the skin, the nails, and the hair, and generally any material keratin.

Since the cosmetic compositions according to the invention are intended to be applied topically to the face, neck, hair, nails or any other cutaneous zone of the body, they are in a form suitable for such a topical application. Thus, the compositions of the invention are in the form of cream, ointment, emulsion, gel, lotion or serum. These compositions may constitute protective creams, treatment or care for the face, for the hands or for the body, protective body care milks, lotions, gels or foams for the care of the skin.

The compositions according to the invention may be in the form of anti-aging compositions, anti-stress compositions, in particular anti-oxidative stress, anti-radical compositions, cell anti-aging compositions, antioxidant compositions, anti-wrinkle compositions, compositions promoting the elasticity of the skin or its barrier function. They are especially in the form of aqueous solutions or lotions, hydroalcoholic or oily, dispersions of the lotion or serum type, anhydrous or oily gels, emulsions of liquid or semi-liquid consistency of the milk type, obtained by dispersion of a fatty phase in an aqueous phase (O / W) or conversely (W / O), suspensions or emulsions of soft, semisolid or solid consistency of the cream, gel, microemulsion or micro-capsule type, micro-particles, or vesicular dispersions of ionic and / or nonionic type and the like and generally any other cosmetic form compatible with the presence and incorporation of such an extract.

Compositions of the invention for topical application suitably comprise a physiologically acceptable carrier. By physiologically acceptable medium is meant in a known manner a medium compatible with the skin, the mucous membranes (including the interior of the eyelids and the lips), the nails and / or the keratinous fibers (hair and eyelashes) and incorporating the extract usable according to the invention. In addition, the compositions of the invention may contain adjuvants customary in the cosmetic field, such as hydrophilic or lipophilic active agents, preservatives, antioxidants, perfumes, fillers, dyestuffs (pigments or dyes), filters solar, solvents and more lipid vesicles. These adjuvants are used in the usual proportions in the cosmetic or dermatological field, and for example from 0.01 to 20% of the total weight of the composition, and they are, according to their nature, introduced into an aqueous phase or in an oily phase. of the composition, or in vesicles.

Of course, those skilled in the art will take care to choose this or these optional additives and / or their amounts in such a way that the advantageous properties intrinsically attached to the composition according to the invention are not, or not substantially, altered by the addition or additions envisaged.

Depending on the fluidity of the composition that it is desired to obtain, one or more gelling agents may be added thereto. As gelling agents, mention may be made of clays, polysaccharide gums and their derivatives, in particular xanthan gum, carboxymethylcellulose, hydroxypropyl guar, carboxyvinyl polymers or carbomers, polyacrylamides, such as the product marketed under the name SEPIGEL 305 by the company SEPPIC and polymers acrylamidomethylpropanesulfonic acid at least partially crosslinked such as the product sold under the name HOSTACERIN AMPS by HOECHST. These gelling agents are generally used at concentrations ranging from 0.1 to 10%, preferably from 0.1 to 5% and better still from 0.1 to 3% of the total weight of the composition. According to a preferred embodiment of the invention, the composition has a pH close to that of the skin, of between 4 and 9, preferably between 4 and 7. These compositions are prepared according to the usual methods in the cosmetics field. For example, by mixing the extract with the cosmetically acceptable vehicle, or any other premix. The extract or extracts are used as obtained, or concentrated or diluted. Preferably, the concentration of the extract used in the compositions is less than about 2 mg dry / ml, ie about 10% by weight of extract in solution. Moreover, the final mass concentration of extract in the composition does not exceed 10% by weight. In the following, the concentrations expressed in percentage correspond to the percentage of pure liquid extract resulting from the extraction, the concentrations expressed in mg dry / ml correspond to the mass of the freeze-dried extract (dry) per milliliter of solvent. Thus, the extracts that can be used according to the invention represent between 0.01% and 10% of the total weight of the composition, preferably between 0.1% and 5% of the total weight of the composition.

During the incorporation, preferably, the temperature of the extracts, vehicles, adjuvants, and mixtures is between 40 ° C and 85 ° C, preferably between 40 ° C and 80 ° C. Finally, in the final composition, the ethanol content does not exceed 40% by weight.

As the solvent which can be used in the composition of the invention, mention may be made of any cosmetically acceptable solvent. The compositions according to the invention may further contain any other compound having moisturizing properties and / or improving the barrier and / or antioxidant function and / or stimulating filaggrin. The compositions can in particular comprise other mushroom extracts, the extracts obtained which can be used in a mixture, or other species, or any other product leading in particular to a modulation of the glycation, the formation of filaggrin or the presence of radicals.

The invention also relates to an anti-aging cosmetic treatment method, that is to say to fight against the signs of skin aging of applying to the skin a composition as defined above.

EXAMPLES Example 1 An extract according to the invention was obtained from a cep dry matter mainly composed of Boletus edulis and Boletus aereus, Boletus pin / cola and Boletus aestivalis, at 3% in water, by extracting at reflux at about 80 ° C for about 30 minutes The extract was recovered by centrifugation at 3500 rpm for 10 minutes. The extract obtained has a solids content of about 19.34 g / l, determined by lyophilization of the material; a pH of between 4 and 6, pH determined by a potentiometric method. a protein level of 0.02 g / L, determined by a colorimetric assay according to the Bradford method [1]; a total sugar level of approximately 7.99 g / L, determined by a colorimetric assay according to the method of Dubois [2] and Blumenkrantz [3]; a total phenol content of approximately 0.85 g / L, determined by a colorimetric assay according to the method of Singleton and Rossi [4]; an oligosaccharide fraction isolated by steric exclusion chromatography on a BIO-RAD CL4B gel, comprised between 70 and 90% by weight of the total carbohydrate fraction. This fraction was composed of 75 to 98 mol% of glucose and included some traces of rhamnose and fucose, a composition determined by gas chromatography equipped with a capillary column (0.32mm * 50m) CP-SIL-5CB and a flame ionization detector (FID). These results were obtained by trimethylsilylation of monosaccharides according to the Kamerling method [5]; a polysaccharide fraction isolated by steric exclusion chromatography on a BIO-RAD CL4B gel representing between 5 and 30% by weight of the total carbohydrate fraction. It consisted of at least 60% glucose and 20 to 30% mannose, and included some traces of xylose and galactose, a composition determined by gas chromatography. This fraction consists essentially of glucans having a molecular mass of between 1000 and 50000 Da. The extract can be taken up in various concentrations in water, as will become apparent in the following examples, depending on the tests carried out.

EXAMPLE 2 Effect on Stimulation of Filaggrin Synthesis 1. MATERIAL AND METHODS 1.1 Biological Models. Normal Human Epidermal Keratinocytes (NHEK) Cell Type: NHEK, KO74 Used at 3rd Pass Culture Conditions: 37 ° C, 5% CO2 20 - Culture medium: Keratinocyte-SFM (Invitrogen 17005-034) supplemented with Epidermal Growth Factor (EGF) 0.25 ng / ml - Pituitary extract (EP) 25 ng / ml (Invitrogen 3700015) - Gentamycin 25 pg / ml (Sigma G1397) - Test medium: Keratinocyte-SFM (Invitrogen 17005-034) supplemented with Gentamycin 25 μg / ml (Sigma G1397). 25. Normal human epidermal fibroblasts (NHDF) - Cell type: NHDF, PF2 pool used in the 5th pass Culture conditions: 37 ° C, 5% CO2 - Culture medium: DMEM (Dulbecco's modified Eagle's medium) (Invitrogen 21969035) 30 Glutamine 2 mM (Invitrogen 25030024) Penicillin 50 IU / ml - Streptomycin 50 μg / ml (Invitrogen 15070063) Fetal calf serum (FCS) 10% (Invitrogen 10270098) .2 Compounds tested and references Compound tested Aspect Stock solution Dilution Concentrations tested Ex-extract . 1 Liquid / Medium 1%, 5% and 10% Storage under test (0.19 and 1.93 mg dry / ml) on ambient keratinocytes Reference Stock Solution Dilution Concentration Tested Calcium Chloride 150 mM Medium 1.5 mM (PROLABO 22319294) Retinoic acid 10-2 M DMSO Medium 10 'M (Sigma R2625) assay 1.3 Preliminary cytotoxicity tests Keratinocytes Fibroblasts Plates: 96 wells Cell / well: 20000 cells / well in 2000 cells / well medium test medium Product range: cf. Table 1 cf. Table 2 Replicates: 6 Contact 48 hours + 96 hours 72 hours Evaluation parameter: reduction of MTT and morphological observations under the microscope (objective x 10) 1.4 Evaluation of the expression of filaggrin Normal human epidermal keratinocytes were cultured in plates of 96 wells in culture medium for 24 hours. Subconfluently, the culture medium was replaced with test medium containing or not (control) test compounds or references (retinoic acid and CaCl2). All conditions were realized in n = 6. After 144 hours of incubation, the culture medium was removed and the cells were rinsed and fixed with a solution of PBS containing 4% paraformaldehyde and then permeabilized with a solution of PBS containing 0.1% triton X-100. Non-specific binding sites were then saturated by incubation in PBS / TWEEN / 5% BSA buffer. The mats were labeled with the primary antibody directed against the protein of interest (FLG, mouse monoclonal antibody, Tebu sc-66192) and then revealed by a secondary antibody coupled to a fluorochrome and directed against mouse immunoglobulins (GAM- Alexa 488, Invitrogen A11001). In parallel, the nuclei of the cells were stained with Hoechst (bis-benzimide, Sigma B1155). Acquisition of the images was performed using the ln Cell AnalyzerTM 1000 (GE Healthcare). Controls without primary antibody (secondary antibody alone) were performed to adjust the acquisition parameters of the camera. For each well, 5 captures of digitized images were made. The markings were quantified by measuring the fluorescence intensity of the proteins relative to the number of nuclei identified by the Hoechst (digital data integration by the Developer Toolbox 1.5 software, GE Healthcare).

1. 5 Data processing The raw data has been transferred and processed using Microsoft Excel® software. Intergroup comparisons were performed using the Student's test. Statistical analyzes can be interpreted if n? 5; and for n <5, the calculated data are provided for information only. Formulas used in this report:

Percentage of stimulation: Average of values - Mean of control 0/0 stimulation = X 100 Average of indicator Mean error of mean: esm = standard deviation (Sd) / end The standard error of mean (esm) represents the deviation of the sample mean from the average of the true population. The esm is calculated by dividing the Sd by the square root of the sample size

2. RESULTS 2.1 Preliminary cytotoxicity tests The effect of the extract of Example 1, at various concentrations, on the viability of the keratinocyte cultures and on the viability of the fibroblast cultures are respectively presented in Table 1 and in Table 2. .

These tests make it possible to determine the maximum non-cytotoxic concentration of the extract for keratinocytes and fibroblasts: 10% (1.93 mg dry / ml). Table 1 Extract of Example 1 Unit% 0.25 0.50 1.00 5.00 10.00 20.00 50.00 100.00 * 0.04 0.09 0.19 0.96 1.93 3.86 9.65 19.34 Witness 103 100 95 100 97 94 88 75 68 11 109 99 98 93 93 93 84 73 69 21 105 90 91 90 92 92 82 70 70 22 Sustainability (%) 104 87 89 92 88 93 79 72 67 24 103 92 95 93 96 93 79 74 70 7 112 97 102 106 104 105 88 80 80 29 Average 100 95 96 95 95 83 74 71 19 esm 2 2 2 2 2 2 1 2 3 Observations + + + + + , ag +/-, ag -, ag -, ag -, ag morphological +: normal population; + 1-: growth reduction; -: toxicity; 0: cell death g: grains of compound; op: opacity due to the compound. *: morphological changes; ag: agglutinated cells esm: standard error of mean (standard deviation divided by square root of sample size) concentration in mg of dry matter per ml Table 2 Extract of Example 1 Unit% 0.25 0.50 1.00 5.00 10.00 20.00 50.00 100.00 * 0.04 0.09 0.19 0.96 1.93 3.86 9.65 19.34 Witness 99 91 104 92 84 76 71 63 48 20 102 101 101 96 87 73 81 67 53 22 Sustainability (%) 104 94 98 97 92 84 81 72 51 24 98 96 95 97 91 83 83 69 54 22 104 102 103 102 99 85 81 74 49 22 103 104 101 98 94 82 77 65 46 23 Average 100 100 97 91 80 79 68 50 22 esm 1 1 1 2 2 2 2 1 1 Observations + + + + + + +, g +/-, g -, g Morphological +: normal population; + 1-: growth reduction; -: toxicity; 0: cell death g: grains of compound; op: opacity due to the compound; *: morphological changes; ag: agglutinated cells esm: standard error of mean (standard deviation divided by square root of sample size) * concentration in mg of dry matter per ml Results of MTT viability tests (3- (4) 5-dimethylthiazol-2-yl) -2,5 diphenyl-tetrazolium bromide) and the observation of the cell mats led to the selection of the concentrations to be tested in the rest of the test (see §1.2). 2.2 Effect on the expression of filaggrin.

The effect of the extract of Example 1, at different concentrations, on the expression of filaggrin in keratinocyte cultures is presented in Table 5 below. Table 3 Treatment Concentration Intensity of% control esm (%) P fluorescence / number of cells (AU) control 338 964 1122 100 25 - 1220 2483 950 Acid 294 retinoic 10- 'M 18 15 5 4 ** 23 14 Chloride of 9410 Calcium 1.5 m M 6526 12886 682 131 ** 11348 4681 3409 4154 1% 1815 or 6035 304 49 0.19 mg 3177 ** dry / ml 3443 Excerpt from ex. 2891 1 1170 5% 3960 or 4607 309 69 0.96 mg 3841 sec / ml 6620 1643 5209 10% 2973 or 5138 801 260 1.93 mg dry 10802 / ml 23639 8926 (1) Threshold of statistical significance: ns:> 0.05 , Not significant *: 0.01 to 0.05, Significant 10 **: 0.001 to 0.01, Very significant ***: <0.001, Extremely significant The reference retinoic acid, tested at 10 'M, significantly decreased the expression of filaggrin ( 5% relative to the control, p <0.01). The reference calcium chloride, tested at 1.5 mM, significantly increased the expression of filaggrin (682% relative to the control, p <0.01). These results were expected and validated the test.

The extract of Example 1, tested at these concentrations 1%, 5% and 10% significantly increased the expression of filaggrin in a concentration-dependent manner (304% relative to the control, p <0.01, 309% relative to to the control, p <0.05 and 801% relative to the control, p <0.05).

In conclusion, the extract of Example 1 exhibited a pro-differentiating effect (increased expression of filaggrin).

EXAMPLE 3 Anti-Radical Effect: The activity of free radical scavenger was evaluated by the DPPH test (1,1-diphenyl-2-picrylhydrazyl) according to J. Buenger [7] and Yoshida [8]. The result of this test is expressed in% free radical scavenging. DPPH is a stable radical due to the delocalization of a free electron around the molecule. This single electron produces a purple coloration of DPPH characterized by an absorption band centered at 520 nm. When the DPPH is mixed with a substance capable of giving a hydrogen atom, the DPPH passes in reduced form and the color disappears. Ascorbic acid is used as a positive control. 2 ml of extract of Example 1 or positive control are added to 2 ml of DPPH at 50 μM, the absorbance is measured at 515 nm after 30 minutes. The percent inhibition is given by the following equation: DPPH reduction%: Q (% inhibition) = (Ao-Ae) / Ao Ao: reference absorbance Ae: sample absorbance Concentration% inhibition Control + 1% 95.65 Extract 1% 0.19 mg dry / ml 84.15 5% 0.95 mg dry / ml 77.28 10% 1.9 mg dry / ml 68.72 The results show that the active ingredient at 1% inhibits 84% of radicals free present.

EXAMPLE 4 Anti-Glycation Effect: The glycation phenomenon is represented in this test by bringing into contact a protein, BSA (Bovine Serum Albumin), and a reducing sugar, glucose, thus making it possible to translate the glycation reaction. Aminoguanidine chloride is used as a positive control. The evaluation of glycation inhibition was performed according to Xiaofang Peng et al. [9], and Rahbar et al. [10]. 5 g of BSA and 14.4 g of D-glucose (800 mmol / L) are dissolved in 100 ml of phosphate buffer (1.5 M, pH 7.4) containing 0.2 g / l of NaN 3 to obtain a solution. baseline with 50 mg / mL BSA and 0.8 mol / L D-glucose (144 mg / mL). 2 ml of the reference solution is incubated at 37 ° C. for 7 days in the presence or absence of 1 ml of extract of Example 1 in a phosphate buffer (1.5 M, pH 7.4). The solution also contains 0.2 g / L NaN3 to ensure the aseptic condition. 100 mM aminoguanidine chloride is used as a positive control. The mixture is incubated for 7 days at 37 ° C. The fluorescent intensity is measured at an emission length of 410 nm and excitation of 330 nm. The percentage inhibition of the glycation products is given by the equation:% inhibition = [1 - (fluorescence with inhibitor / fluorescence without inhibitor)] * 100 Concentration% inhibition Control + 1% 80.43 Extract 1% 0.19 mg dry / ml 3.60 5% 0.95 mg dry / ml 23.30 10% 1.9 mg dry / ml 43.90 The results show that the 10% active principle inhibits 44% of the glycation products.

EXAMPLE 5 Cosmetic Composition Composition% wt 3-polyglyceryl methylglucose distearate emulsifier 1,500 cetearyl alcohol 3,000 PEG-40 hydrogenated castor oil 3,000 sodium cetearyl sulphate 3,000 Stabilizer glyceryl stearate 4,000 emulsifier isononyl isononanoate 8,000 Aqua water 75,495 preservative methylchloroisothiazolinone / 0.005 methylisothiazoline Active Extract Bolet of Example 1 2.000 * pure active agent derived from extraction Example 6: cosmetic composition Composition% by weight Aqua water 80.977 Acrylate gelling agent / C1o30 alkylacrylate crosspolymer 0.475 Chondrus crispus (carrageenan) 0.163 Betaine wetting agent 0.400 Humectant glycerin 4.000 Solvent polyethylene glycol-8 0.500 Preservative Chlophenesin 0.171 methylparaben 0.057 phenoxyethanol 0.587 butylparaben 0.684 Triclosan 0.050 sodium hydroxide 0.050 Emoliant propylene glycol 0.313 isononylisononanoate 6.000 caprylic / capric triglyceride 4.000 Stabilizer cetyl alcohol 0.600 Oil B Hydroxytoluene 1,500 Gossypium herbaceum - Seed Oil 1,500 Cotton Active * Boletin Extract of Example 1 2,000 18 * Pure Active Extracting Product REFERENCES [1] Bradford, A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Analytical Biochemistry 72, 248-254 (1976). [2] Dubois, A colorimetric method for the determination of sugars, Nature 168: 167, (1951). [3] Blumenkrantz and Asboe-Hansen, New method for quantitative determination of uronic acids, Anal Biochem 54: 484, (1973). [4] Singleton and Rossi, Colorimetry of total phenolics with phosphomolybdicphosphotungstic acid reagents, AJEV, 16: 144-158, (1965). [5] Kamerling J.P. et al., Characterization by Gas-liquid chromatography- Mass spectroscopy and proton magnetic- Resonance spectroscopy of pertrimethylsilyl of glycoproteins and glycopeptides. Biochem. J, 151, 491-495, (1975). [6] Christophe Chauveau et al., A water-soluble 8-D-glucan from Boletus erythropus, Phytochemistry, Vol. 43, No. 2, pp. 413-415, (1996). [7] J. Buenger et al., "An interlaboratory comparison of methods used to assess antioxidant potentials", Internationnal Journal of Cosmetic Science, 28, 135-146, (2006). [8] Yoshida, T. et al., "Studies of Inhibition Mechanism of Autoxidation by Tannins and Flavonoids." Radical-Scavenging Effects of Tannins and Related Polyphenols on 1,1-Diphenyl-2-picrylhydrazyl radical., Chem Pharm. 37 (7), 1919-1921 (1989) [9] Xiaofang Peng et al., "Inhibitory effect of mung bean extract and its constituents vitexin and isovitexin on the formation of advanced glycation endproducts", Food Chemistry 106, 475- 481, (2008). [10] Samuel Rahbar and James L. Figarola, "Novel Inhibitors of Advanced Glycation Endproducts," Archives of Biochemistry and Biophysics 419, 63-79, (2003). [11] Tsai Shu-Yao et al "Antioxidant properties of Agaricus blazei, Agrocybe cylindracea, and Boletus edulis", Lebensmittel - Wissenschaft + Technology, Vol 40, No. 8, pp. 1392-1402 (2007), Elsevier.

Claims (13)

REVENDICATIONS1. Cosmetic use of a pedunculated edible mushroom extract, in particular a cep or boletus extract, for the anti-aging treatment of the skin by application of a topical composition comprising at least one such extract and a cosmetically acceptable vehicle, in particular with anti-radical, anti-glycation and / or stimulation of the synthesis of filaggrin. 2. Cosmetic use according to claim 1, wherein the extract is an aqueous extract. 3. Cosmetic use according to claim 1 or 2, wherein the mushroom extract meets at least one of the following criteria: a) dry matter content, determined by lyophilization, between 0.1 and 100 g / L, of preferably between 1 and 50; b) pH of between 3 and 8, preferably between 4 and 6; c) protein level between 0.0005 and 10 g / L, preferably 0.001 and 1 g / L; d) total sugar levels between 0.01 and 100g / L, preferably between 0.05 and 50g / L; e) total phenol content between 0.001 and 50 g / l, preferably 0.005 and 10 g / l; f) an oligosaccharide fraction representing between 50 and 95% by weight of the total carbohydrate fraction, preferably composed of 65 to 99 mol% of glucose, preferably 70 and 90% by weight of the total carbohydrate fraction, preferably composed of 75 to 98 mol% of glucose and optionally comprising traces of rhamnose and fucose; g) a polysaccharide fraction representing between 2 and 50% by weight of the total carbohydrate fraction, preferably comprising at least 50% glucose and 15 to 35% mannose and optionally comprising traces of xylose and galactose, preferably representing and 30% by weight of the total carbohydrate fraction, preferably comprising at least 60% deglucose and 20 to 30% mannose and optionally comprising traces of xylose and galactose; h) a polysaccharide fraction consisting essentially of glucans with a molecular mass of between 500 and 65,000 Da. 4. The cosmetic use according to any of claims 1 to 3, wherein the aqueous mushroom extract meets at least one of the following criteria: a) solids content, determined by lyophilization, between 15 and 25 g / L, preferably 19.34 ± 0.20 g / L; b) pH of between 3 and 8, preferably between 4 and 6; c) protein level of 0.02 ± 0.01 g / L; d) total sugar levels ranging from 1 to 25 g / L, preferably 7.99 ± 1 g / L; E) total phenol levels of 0.1 to 2.5 g / L, preferably 0.85 ± 0.05 g / L; f) an oligosaccharide fraction, isolated by steric exclusion chromatography on BIO-RAD CL4B gel, representing between 70 and 90% by weight of the total carbohydrate fraction composed of 75 to 98 mol% of glucose and comprising some traces of rhamnose and of fucose; g) a polysaccharide fraction representing between 5 and 30% by weight of the total carbohydrate fraction, comprising at least 60% of glucose and 20 to 30% of mannose, and possibly traces of xylose and galactose; h) a polysaccharide fraction consisting essentially of glucans with a molecular mass of between 1000 and 50 000 Da. 30 5. Cosmetic use according to one of claims 1 to 4, characterized in that the mushroom extract is selected from the group of extracts of Boletus erythropus, Boletus edulis, Boletus badius, Boletus luridus, Suillus bovinus, Suillus granulatus, Leccinum scabrum, Boletus aereus, Boletus pinicola and Boletus aestivalis or mixtures thereof and in particular Boletus edulis, Boletusaereus, Boletus pinicola and Boletus aestivalis, more particularly Boletus edulis. 6. Cosmetic use according to one of claims 1 to 5, characterized in that it comprises from 0.01 to 10% by weight of the extract or extracts, preferably from 0.1 to 5% by weight of the extract or extracts, relative to the total weight of the composition. 7. Use according to one of claims 1 to 6, characterized in that the concentration of the extract used is less than or equal to about 2 mg of dry matter per ml. 8. Cosmetic use according to one of claims 1 to 7, in the form of anti-aging composition, anti-stress composition including anti-oxidative stress, anti-radical composition, antioxidant composition, anti-wrinkle composition, anti-aging composition. cellular aging, composition promoting the elasticity of the skin or its barrier function. 9. Topical cosmetic use of a pedunculate edible mushroom extract in a cosmetically acceptable vehicle, especially a cep or boletus extract, more particularly an extract of Boletus edulis, to stimulate the synthesis of filaggrin. 10. A topically applied cosmetic composition comprising a cosmetically acceptable vehicle and at least one edible pedunculate fungus extract meeting the following criteria: a) dry matter content, determined by lyophilization, between 0.1 and 100 g / L, of preferably between 1 and 50; b) pH of between 3 and 8, preferably between 4 and 6; c) protein level between 0.0005 and 10 g / L, preferably 0.001 and 1 g / L; d) total sugar content between 0.01 and 100 g / l, preferably between 0.05 and 50 g / l, e) total phenol content between 0.001 and 50 g / l, preferably 0.005 and 10 g / l; f) an oligosaccharide fraction representing between 50 and 95% by weight of the total carbohydrate fraction, preferably composed of 65 to 99 mol% of glucose, preferably 70 to 90% by weight of the total carbohydrate fraction, preferably composed of 75 to 98 mol% of glucose and optionally comprising traces of rhamnose and fucose; g) a polysaccharide fraction representing between 2 and 50% by weight of the total carbohydrate fraction, preferably comprising at least 50% glucose and 15 to 35% mannose and optionally comprising traces of xylose and galactose, preferably representing between 5 and 30% by weight of the total carbohydrate fraction, preferably comprising at least 60% glucose and 20 to 30% mannose and optionally comprising traces of xylose and galactose; h) a polysaccharide fraction consisting essentially of glucans with a molecular mass of between 500 and 65,000 Da. 20 11. Composition according to claim 10, characterized in that the extract meets the following criteria: a) dry matter content, determined by lyophilization, of between 15 and 25 g / L, preferably 19.34 ± 0.20 g. / L; b) pH of between 3 and 8, preferably between 4 and 6; C) protein level of 0.02 ± 0.01 g / L; d) total sugar levels ranging from 1 to 25 g / L, preferably 7.99 ± 1 g / L; e) total phenol content of between 0.1 and 2.5 g / l, preferably 0.85 ± 0.05 g / l; F) an oligosaccharide fraction, isolated by steric exclusion chromatography on BIO-RAD CL4B gel, representing between 70 and 90% by weight of the total carbohydrate fraction composed of 75 to 98 mol% of glucose and comprising some traces of rhamnose and fucose; G) a polysaccharide fraction representing between 5 and 30% by weight of the total carbohydrate fraction, comprising at least 60% of glucose and 20 to 30% of mannose, and possibly traces of xylose and galactose; h) a fraction essentially consisting of glucans with a molecular mass of between 1000 and 50 000 Da. 12. Composition according to claim 10 or 11, characterized in that the extract is selected from the group of extracts of Boletus erythropus, Boletus edulis, Boletus badius, Boletus luridus, Suillus bovinus, Suillus granulatus, Leccinum scabrum, Boletus aereus, Boletus pinicola and Boletus aestivalis or mixtures thereof and in particular Boletus edulis, Boletus aereus, Boletus pinicola and Boletus aestivalis, more particularly Boletus edulis. 13. Cosmetic process for anti-aging treatment of the skin, in which a composition used in one of Claims 1 to 9 or a composition according to one of Claims 10 to 12 is applied.