FR2971711A1 - Use of extract of Einkorn wheat (Triticum monococcum), as active agent for activating synthesis of extracellular matrix protein of skin, in cosmetic composition comprising medium for fighting against the appearance of signs of skin aging - Google Patents

Abstract

Use of an extract of Einkorn wheat ( Triticum monococcum) (I), as active agent for activating synthesis of the extracellular matrix protein of the skin, in a cosmetic composition comprising a medium for fighting against the appearance of signs of skin aging originating from the degradation of the extracellular matrix, is claimed. An independent claim is included for an extract of Einkorn wheat ( Triticum monococcum) obtained by contacting crushed seeds or flour of (I) Einkorn wheat ( Triticum monococcum) with an aqueous solution, under agitation, successively hydrolyzing the mixture using proteases and then purifying by filtration, and separating low molecular weight compounds by ultrafiltration. ACTIVITY : Dermatological. MECHANISM OF ACTION : Extracellular matrix protein activator; Collagen I synthesis stimulator; Collagen III synthesis stimulator; Fibronectin synthesis stimulator.

Description

Field of the Invention The invention is in the field of cosmetics and active ingredients for cosmetics. The present invention relates to an extract of small spelled (Triticum monococcum) obtained by hydrolysis of flour or spelled seeds using at least two proteans other than papain. The present invention also relates to a cosmetic composition comprising said small spelled extract. The present invention also relates to the cosmetic use of a small spelled extract (Triticum monococcum) as an activating agent for the expression of extracellular matrix proteins. The active agent can be used alone or in combination with other active agents. The invention also relates to the use of a cosmetic composition comprising a small spelled extract for increasing the expression of the proteins of the extracellular matrix, delaying the onset and / or combating the cutaneous signs of aging linked to the degradation of extracellular matrix, and in particular wrinkles, sagging and loss of skin volume and dehydration of the skin and finally, cosmetic care methods intended to delay the appearance and / or treat the cutaneous signs of aging.

BACKGROUND OF THE INVENTION The skin is a covering organ composed of several layers (dermis, dermal-epidermal junction, epidermis). The dermis is the skin support tissue and consists of water, elastin fibers and collagen fibers (70% of dermal fibers), wrapped in an interstitial matrix of proteoglycans or glycosaminoglycans.

Fibroblasts are the main cellular component of the dermis and are responsible for the synthesis of collagen fibers and elastin fibers. Glycosaminoglycans provide structuring of collagen and elastin fibrils and storage of water, thanks to their exceptional hygroscopicity. Hyaluronic acid is the most abundant glycosaminoglycan in the skin, and the major constituent of the dermis but is also present around keratinocytes in the epidermis. The glycosaminoglycan and collagen complexes are major players in the suppleness and firmness of the skin. The skin, like all other organs, is subject to the complex physiological process of aging. We distinguish between intrinsic or chronological aging and extrinsic aging. Intrinsic aging is the consequence of genetically programmed senescence and biochemical alterations due to endogenous factors. At the level of the skin, it is characterized by a slowing of the regeneration of cells and extracellular matrices, resulting in a decrease in the thickness of the dermis, collagen fibers that are less numerous and disorganized, and an altered structure of the elastic fibers of the dermis. papillary, having as clinical and functional consequences a dryness, a laxity of the skin and the appearance of fine lines and wrinkles. 10 Extrinsic aging, for its part, is due to environmental aggressions such as pollution, sun, diseases, lifestyle, etc. Hydrolytic extracts of small spelled and their cosmetic uses to provide antioxidant or protective effects vis-à-vis external aggression and thus fight against extrinsic photo-induced aging have already been described in FR2873918 and FR2925332 patents. The inventors have now demonstrated that such extracts of small spelled were good activating agents for the synthesis of extracellular matrix proteins and could be used as new active agents for combating the appearance of cutaneous signs of aging originating from degradation of the extracellular matrix. The inventors have also found that new extracts of small spelled, obtained by hydrolysis using at least two proteases different from papain, had the same properties on the skin, while having the advantage of being more homogeneous, more tables, and presenting less risk of allergic reactions in dermato-cosmetic. SUMMARY OF THE INVENTION The inventors have demonstrated that extracts of small spelled are good activating agents for the synthesis of extracellular matrix proteins. As a result, these extracts are adapted to fight the cutaneous signs of aging originating from the degradation of the extracellular matrix. The extracts according to the invention are characterized by the fact that they: - increase the expression of type I and III collagens - increase the expression of fibronectin, - increase the expression of hyaluronic acid 2971711 -3 - - reduce signs of fibroblast senescence The term "spelled extract" or "active activating agent of extracellular matrix proteins" means any extract capable of increasing the amount of collagen, fibronectin and hyaluronic acid present. in the cell, or secreted either by increasing protein synthesis by direct or indirect modulation of gene expression, or by other biological processes such as stabilization of the protein or the stabilization of messenger RNA transcripts. The term "extract" refers to any substance or mixture of substances, or isolated preparation, obtained after hydrolysis of plant material. The term "signs of fibroblast senescence" refers to the expression profile of biochemical markers associated with the cellular senescence phenotype. By skin is meant all the covering tissues constituting the skin, the mucous membranes and the integuments, including the hair, the eyelashes and the eyebrows. Thus, the first object of the invention is the use of a small spelled extract (Triticum monococcum), as an activating active agent for the synthesis of extracellular matrix proteins of the skin, in a cosmetic composition. The cosmetic composition further comprises a physiologically adapted medium. More particularly, small spelled extract is used as an active agent to increase the synthesis of collagens I and III and fibronectin by fibroblasts of human skin. Thus, according to this embodiment of the invention, the small spelled extract is used as an active agent to increase the density of the dermis, and thus the firmness of the skin, to retard or reduce the relaxation of the lines. of the face, loss of dermal volume, thinning of the skin, atony, fine lines, deep wrinkles and dermal atrophy. The said small spelled extract is still used as an active agent to increase the expression of hyaluronic acid, both by fibroblasts and by keratinocytes. According to this particular embodiment of the invention, said small spelled extract is used to increase the ability of all layers of the skin to retain water and delay or reduce the dehydration of the skin associated with intrinsic aging and particularly to decrease the amount of glycosaminoglycans in the extracellular matrices. According to another embodiment of the invention, said small spelled extract is still used as an active agent to delay the onset or decrease senescence of skin cells. The term "delaying the onset or decreasing senescence of dermal cells" is intended to mean that, according to this advantageous aspect of the invention, the extract of the invention decreases the expression of senescence markers in fibroblasts. humans whose senescence was induced by extinction of the FOXO3a gene.

The active agent according to the invention is an aqueous extract of spelled and more particularly an aqueous extract of seeds of small spelled or engrain (Triticum monococcum). The small spelled is a diploid wheat, very old, containing a particularly high rate in proteins (Vallega 1992) but also in carotenoids, phosphorus and potassium (Abdel-Aal et al., 1995, Mtuz et al., 2000). In addition, T. monococcum proteins appear to contain little gluten, causing celiac disease (Favret et al., 1984, 1987). Any method of extraction or purification known to those skilled in the art can be used to prepare the extract according to the invention.

According to a particular form of the invention, this extract can be obtained by extraction of proteins of plant origin, followed by controlled hydrolysis which liberates biologically active peptide fragments. Many proteins found in plants are likely to contain biologically active peptide fragments within their structure. The controlled hydrolysis makes it possible to release these peptide fragments. It is possible, but not necessary to carry out the invention, to extract either the relevant proteins first and then hydrolyze them, or to hydrolyze first on a crude extract and purify the peptide fragments. then. In a first step, the seeds are crushed using a plant grinder. The powder thus obtained may subsequently be "delipidated" with the aid of a conventional organic solvent (for example an alcohol, hexane or acetone). According to the invention, the seeds used are not subjected to prior fermentation. Advantageously, the extract according to the invention is made from complete or refined flours derived from these seeds. The proteins of the plant are then extracted according to the conventional method (Osborne, 1924) modified; the seed mill or flour is suspended in about 10 volumes of an aqueous solution and the soluble fraction, containing proteins, carbohydrates and optionally lipids, is collected after centrifugation and drying steps. filtration. The extraction temperature is between 4 and 80 ° C; preferably the operation will be performed cold. This crude solution is then hydrolyzed under mild conditions to generate soluble peptide compounds. Hydrolysis is defined as a chemical reaction involving the cleavage of a molecule by water, this reaction being possible in a neutral, acidic or basic medium. According to the invention, the hydrolysis is advantageously carried out chemically and / or advantageously by proteolytic enzymes. We can then mention the use of amylases and endoproteases of plant origin (papain, bromelain, ficin) and microorganisms (Aspergillus, Rhizopus, Bacillus, Alcalase®, etc.). After filtration, the extract is purified either by precipitation of the molecules of high molecular weight by the addition of alcohol or salts or by filtration. During this filtration the retentate is removed and the filtrate (dialysate) is preserved. At the end of the extraction, the extract obtained is an aqueous solution containing at least 5 g / L of nitrogenous material and comprising peptide compounds of molecular weight less than 15 kDa. The extract is then diluted in water or in any solvent mixture containing water and sterilized by ultrafiltration. The extract is advantageously diluted in one or more physiologically suitable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. By "physiologically fit" it is meant that the selected solvent is suitable for contacting the skin without causing toxicity or intolerance reactions. Thus, after dilution, the small spelled extract has a peptide compound content of between 1.5 and 3.5 g / l. The small spelled extract obtained according to the invention is analyzed qualitatively and quantitatively for its physico-chemical characteristics and its content of peptide compounds. The term "peptide compounds" means the protein fragments, the peptides and the free amino acids present in the mixture. The peptides, amino acids and protein fragments are determined according to conventional techniques, which are well known to those skilled in the art. After this dilution step, the extract may be encapsulated or included in a cosmetic vector such as liposomes or any other microcapsule used in the field of cosmetics or adsorbed on powdery organic polymers, inorganic supports such as talcs and bentonites, and more generally solubilized in, or attached to, any physiologically adapted vector.

According to a particularly advantageous form of the present invention, the small spelled extract is obtained by a process employing at least two proteases other than papain: It is thus possible to prepare a small spelled extract containing a level of compounds. peptides higher than the dry weight and whose peptide compounds have a molecular weight of less than 5 kDa, performing on the one hand a hydrolysis by combination of several proteases and on the other hand by purifying the soluble compounds obtained by filtrations and ultrafiltration on membranes with a low cut-off. The use of peptide hydrolysates of low molecular weight, and concentrated in peptide compounds compared with extracts of the prior art, has many advantages in cosmetics. In addition to generating peptide compounds that did not pre-exist in the starting protein mixture, the hydrolysis and purification described in the process below make it possible to obtain more stable, easier to standardize and less risky mixtures. allergic reactions in dermatocosmetics. According to this particularly advantageous form, which is the subject of the invention, the small spelled extract is obtained by the following process: a) Extraction by contacting crushed seeds or small spelled flour (Triticum monococcum) with an aqueous solution with stirring, b) hydrolyzed using at least two proteases, c) purification by at least one filtration step, d) separation of low molecular weight compounds by ultrafiltration. More particularly, the hydrolysis can be carried out using the proteases bromelain and Alcalase®. The joint action of these two proteases makes it possible to generate peptide fragments that are smaller and of a different nature than those generated during the action by papain. In fact, although both bromelain and papain are endoproteases (Cys-protease), the complementary action of the alkalase O, which is an exoprotease, allows the hydrolysis of proteins by other proteins. cleavage sites, thus leading to peptides or peptide compounds of nature and different from those obtained by hydrolysis with papain. The polyacrylamide gel electrophoresis analysis showed a band at 24 kDa, corresponding to the inactivated enzymes, and then peptides with a molecular weight of less than 5 kDa. The distribution profile of the peptide compounds of the extract obtained according to the invention is completely different from that of an extract obtained for example by hydrolysis using papain. According to a preferred form of the invention, the extract is purified in step d) by ultrafiltration on filters having a cut-off of 5 kDa, so as to eliminate all compounds of molecular weight greater than 5 kDa. Polyacrylamide gel electrophoresis analysis shows that all peptide compounds have a molecular weight of less than 5 kDa.

The small spelled extract according to the invention is diluted in water or in any mixture containing water, and the diluted extract is sterilized by ultrafiltration. The extract according to the invention is advantageously diluted in one or more physiologically suitable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. "Physiologically suitable" means that the chosen solvent is suitable for coming into contact with the skin without causing toxicity or intolerance reactions. Thus, after dilution, the small spelled extract has a peptide compound content of between 1.5 and 3.5 g / l.

The cosmetic compositions used according to the invention contain the extract of small spelled as active activating agent at a concentration of between 0.0001% and 20% approximately, and preferably at a concentration of between 0.05% and 5%. about% relative to the total weight of the final composition. This range of concentrations represents the necessary amount of active agent to obtain the desired molecular effect, namely, to activate the expression of type I and III collagens, fibronectin and hyaluronic acid. Preferably, the composition used according to the invention is in a form suitable for topical application comprising a physiologically suitable medium for the skin. By "physiologically suitable" is meant media suitable for use in contact with human skin or integuments, without risk of toxicity, incompatibility, instability, allergic response and other side effects. The term "topical application" means applying or spreading the active agent according to the invention, or a composition containing it, on the surface of the skin. These compositions may especially be in the form of an aqueous solution, hydro-alcoholic or oily, an oil-in-water emulsion, water-in-oil or multiple emulsions, aqueous or anhydrous gel, colloid. These compositions may also be in the form of creams, suspensions or powders, suitable for application to the skin, mucous membranes, lips and / or integuments. These compositions may be more or less fluid and have the appearance of a cream, lotion, milk, serum, ointment, cream, paste or paste. a foam. They can also be in solid form, as a stick, or be applied to the skin in the form of an aerosol. They can be used as a care product and / or as a make-up product for the skin. These compositions may also be adapted to applications on the scalp and / or the hair, and in particular be in the form of shampoo, conditioner, hair treatment lotion, hair masks, etc.

All of these compositions additionally comprise any additive commonly used in the intended field of application and the adjuvants necessary for their formulation, such as co-solvents (ethanol, glycerol, benzyl alcohol, humectant, etc.). Thickeners, diluents, emulsifiers, antioxidants, dyes, sunscreens, pigments, fillers, preservatives, perfumes, odor absorbers, essential oils, trace elements, essential fatty acids, surfactants, film-forming polymers, chemical or mineral filters, moisturizing agents or thermal waters, etc. For example, mention may be made of water-soluble polymers of the natural polymer type, such as polysaccharides, or polypeptides, cellulosic derivatives of the methylcellulose or hydroxypropylcellulose type, or else synthetic, polaxamer, carbomer, siloxane, PVA or PVP polymers, and especially the polymers sold by ISP (International Specialty Products). In all cases, those skilled in the art will ensure that these adjuvants and their proportions are chosen in such a way as not to harm the desirable advantageous properties of the composition according to the invention. These adjuvants may, for example, be present at concentrations ranging from 0.01 to 20% of the total weight of the composition. When the composition of the invention is an emulsion, the fatty phase may represent from 5 to 80% by weight and preferably from 5 to 50% by weight relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the composition will be chosen from those conventionally used in the field under consideration. For example, they can be used in a proportion ranging from 0.3 to 30% by weight, based on the total weight of the composition.

According to another form of the invention, the compositions that can be used for the invention will be suitable for oral administration. Thus, the compositions may especially be in the form of tablets, capsules, capsules, chewable pastes, powders to be consumed as such or to be mixed extemporaneously with a liquid, syrup, gels, and any other form known to those skilled in the art. They will contain suitable formulation excipients, such as colorants, sweeteners, flavoring agents, bulking agents, binders, preservatives.

It is understood that the active agent according to the invention can be used alone or in combination with other active agents. Advantageously, the compositions that may be used according to the invention contain, in addition, at least one other active agent intended to enhance the action of the active agent according to the invention, in the field of preventing and improving cutaneous signs. aging, or another active agent to expand the range of properties of the composition considered. The following classes of ingredients may be mentioned in a non-limiting manner: regenerating, anti-aging, anti-wrinkle, soothing, anti-radical, anti-glycation, moisturizing, antibacterial, antifungal, keratolytic, myorelaxant, desquamating, firming agents, Agents stimulating the synthesis of dermal macromolecules or energy metabolism or microcirculation or nail growth or hair growth, agents modulating the differentiation or pigmentation of the epidermis, the agents inhibiting metalloproteinases, or screens or sunscreens. In a more particular embodiment, the composition according to the invention will comprise, in addition to the extract according to the invention, at least one activator compound of cytochrome c and / or at least one hydrating compound, such as an activator compound of the aquaporins and / or - at least one sirtuin activator compound and / or - at least one compound which increases cell adhesion and / or - at least one compound which increases the production of proteins matrices such as collagen, fibronectin, laminin, glycosaminoglycans and / or - at least one modulator compound of the proteasome activity and / or - at least one circadian rhythm modulating compound and / or - at least one compound modulator of the HSP proteins and / or, - at least one compound which increases the cellular energy and / or - at least one compound modulating the pigmentation of the skin and / or - at least one activating compound of the coenzyme Q10 and / or , At least one compound improving the barrier function re, such as an activator compound of the transglutaminase or the HMG-CoA reductase and / or - at least one protecting compound of the mitochondrion. Said above compounds may be of natural origin, such as peptide extracts of plants, or of synthetic origin, such as peptides. Regardless of their functions, the other active agents associated with the active agent according to the invention in the composition may have very diverse chemical structures. Non-limiting mention may be made of peptides, vitamin C and its derivatives, group B vitamins, DHEA (dihydroepiandrosterone), phytosterols, salicylic acid and its derivatives, retinoids, flavonoids and amines. sugars, azoles, metal salts, peptide extracts of plant origin or polymers.

The invention finally relates to a cosmetic care method for delaying and / or treating the cutaneous signs of aging originating from the degradations of the extracellular matrix, characterized in that the skin to be treated is applied topically to the skin to be treated. composition according to the invention. Cutaneous signs of aging means any changes in the external appearance of the skin and integuments due to intrinsic aging and in particular the signs originating in the degradations of the extracellular matrix. Among these signs, it is possible to cite the slowing down of the regeneration of the cells or their entry into senescence, the reduction in the thickness of the dermis, the fewer and disorganized collagen fibers, the decrease in the amount of glycosaminoglycans, alteration of the elastic fibers of the papillary dermis. All its biological modifications have as 5 clinical and functional consequences dry skin, sagging skin, loss of hydration, atony, the appearance of deep wrinkles and fine lines, loss of firmness and tone , dermal atrophy.

Other advantages and features of the invention will appear better on reading the examples given for illustrative and non-limiting purposes.

Example 1 Preparation of an extract of small spelled (Triticum monococcum) The small spelled flour (Triticum monococcum) is dissolved in 10 volumes of water in the presence of 2% of POLYCLARO (polyvinylpyrrolidone - PVPP - insoluble). The mixture is adjusted to a pH of between 6 and 8 with a 1M aqueous sodium hydroxide solution. After adjusting the pH, an amylase (hasidase) is added to the reaction medium. The hydrolysis is obtained after 2 hours of stirring at 50 ° C. A protease (2% papain) is then added, which is left to act for 2 hours with stirring at 50 ° C. The enzymes are then inactivated by heating the solution at 80 ° C for 2 hours. After centrifugation, the supernatant aqueous solution corresponding to a crude extract of small spelled is recovered. The purification process of the crude extract begins with successive filtrations using Seitz-Orion plate filters of decreasing porosity (up to 0.2 μm) in order to obtain a brilliant and clear solution, qualified as Extract 1. At this stage, the extract 1 of small spelled is characterized by a light yellow color and by a dry extract grading 70-75 g / kg, a peptide compound level of 10 to 15 g / l and a sugar level of 60 to 65 g / 1. The protein nature of the extract 1 is demonstrated after analysis by NuPAGEO Bis-Tris Pre-cast polyacrylamide gel electrophoresis (Invitrogen). The small spelled protein extract is heated at 70 ° C for 10 minutes under denaturing reducing conditions in NuPAGEO LDS sample preparation buffer. A solution of NuPAGEO Antioxidant is added to the inner vessel (cathode) to prevent the reduced proteins from reoxidizing during electrophoresis. Protein migration is performed in NuPAGE® MES Migration Buffer with the SeeBlue Plus2 standard as a molecular weight marker. Protein staining is performed using Coomassie® Blue R-250. Under these conditions, a 24 kDa band corresponding to the inactivated enzymes is observed, followed by peptide compounds with a molecular weight of less than 15 kDa. The extract 1 is then purified by ultrafiltration using the Pellicon® 2 Biomax 30 kDa cassette in order to eliminate all traces of enzymes. At the end of the purification, a yellow-orange peptide extract is obtained, which is brilliant and limpid. A dilution phase is carried out to obtain a peptide extract characterized by a content of peptide compounds of 1.5 to 3.5 g / l. This extract corresponds to a form of the active agent that can be used according to the invention.

EXAMPLE 2 Preparation of an extract of small spelled (Triticum monococcum) comprising peptides of low molecular weight The flour of small spelled (Triticum monococcum) is dissolved in 10 15 volumes of water in the presence of 2% of POLYCLAR® (Polyvinylpyrrolidone - PVPP - insoluble). The mixture is adjusted to a pH of between 6 and 8 with a 1M aqueous sodium hydroxide solution. After adjusting the pH, two proteases (bromelain and Alcalase®) are added to the reaction medium. The hydrolysis is obtained after 2 hours of stirring at 50 ° C. The enzymes are then inactivated by heating the solution at 80 ° C for 2 hours. After centrifugation, the supernatant aqueous solution corresponding to a crude extract of small spelled is recovered. The process for purifying the crude extract begins with successive filtrations using Seitz-Orion plate filters of decreasing porosity (up to 0.2 μm) in order to obtain a brilliant and clear solution, qualified as At this stage, the extract 1 of small spelled is characterized by a light yellow color and by a solids content of 20-25 g / kg, a peptide compound content of 10 to 15 g / l and a sugar content of 5 to 10 g / 1. The protein profile of the extract 1 is demonstrated after analysis by NuPAGE® Bis-Tris Pre-cast polyacrylamide gel electrophoresis (Invitrogen). The small spelled protein extract is heated at 70 ° C for 10 minutes under denaturing reducing conditions in NuPAGE® LDS sample preparation buffer. A solution of NuPAGEO Antioxidant is added to the inner vessel (cathode) to prevent the reduced proteins from reoxidizing during electrophoresis. Protein migration is performed in NuPAGEO MES migration buffer with the SeeBlue Plus2 standard as a molecular weight marker. Protein staining is performed using Coomassie Blue R-250. Under these conditions, a 24 kDa band corresponding to the inactivated enzymes is observed, followed by proteins less than 5 kDa. The extract 1 is then purified by ultrafiltration using the Pellicon ™ 2 Biomax 5 kDa cassette in order to eliminate all traces of enzymes. At the end of purification, a yellow-orange, bright and clear peptide extract is obtained, which has a dry weight of between 6 and 10 g / kg, a concentration of peptide compounds of between 4 and 7 g / l and a concentration of sugars between 1 and 3 g / 1. A dilution phase is carried out in order to obtain a peptide extract characterized by a content of peptide compounds of 1.5 to 3.5 g / l. This extract, referred to as extract 2, corresponds to a possible form of the active agent according to the invention. Extract 2 is analyzed by high pressure liquid chromatography (HPLC) using an HP1100 device controlled by the ChemStation software. The column used during the elution of the extract 2 is a Nucleosi10 300-5 C4 MPN (125 x 4 min), allowing to chromatograph proteins having molecular weights of 0.2 to 25 kDa (according to a solvent gradient appropriate). Under these chromatographic conditions, several peptide fractions could be isolated, having a total molecular weight of less than 5 kDa. These various fractions are then analyzed by mass spectrometry to identify their molecular peaks.

EXAMPLE 3 Demonstration of the Activating Effect of Spelled Extract on the Expression of Collagen I in Human Fibroblasts The aim of this study is to determine the influence of spelled extract according to US Pat. Example 1 on the expression of collagen I in human fibroblasts by immunoblotting. Protocol: Human fibroblasts in culture are treated with the small spelled extract according to Example 1 at a final concentration of 1% for 48 hours. Cells are lysed in RIPA buffer (Thermo Scientific) containing protease inhibitors (HaTMT Protease Inhibitor Cocktail and EDTA, Thermo Scientific). The cell suspension is centrifuged at 12000 g for 30 minutes at 4 ° C and the supernatant is recovered. The protein concentration of the supernatant is determined by the BCA Protein Assay Kit (Thermo Scientific). A quantity of 20 μg of protein is deposited on a SDS-PAGE gel in 4-12% gradient and transferred onto a nitrocellulose membrane using the iBlot® Dry Blotting System (Invitrogen). The immunological detection is carried out using a polyclonal rabbit antibody specific for Collagen I (Rockland, Ref: 600-401-103-0.5), then a secondary anti-rabbit antibody coupled to a peroxidase (Immunotech , Ref: 115-036-062). A chemiluminescent revelation solution (SuperSignal West Dura Extended Duration Substrate, Thermo Scientific) is then applied and the images are taken using the Chemi-Imager system (Alpha Innotech Corporation). Results: In the conditions tested, more intense luminescence was observed for the treated samples compared to the control conditions. Conclusions: The small spelled extract according to Example 1 stimulates the expression of collagen I in human dermal fibroblasts.

EXAMPLE 4 Demonstration of the activating effect of the small spelled extract according to Example 2 on the expression of collagen I, collagen III, pro-collagen and hyaluronic acid in biopsies of The purpose of this study is to determine the influence of the small spelled extract according to Example 2 on the expression of the following extracellular matrix proteins: collagen I, collagen III, pro-collagen, fibronectin and hyaluronic acid in the human skin. For this purpose, specific markings were made on human skin samples cultured ex vivo. Protocol: Human skin samples are cultured at the air / liquid interface. The small spelled extract according to Example 2 is applied topically to the samples at final concentrations of 0.5%, 1% or 3%, and then the samples are incubated for 24, 48 and / or 72 hours at 37 ° C. ° C. These skin samples are then fixed with formaldehyde then embedded in paraffin or fixed and included in OCT and then frozen at -20 ° C. Sections of 4 μm thick are then made (6 μm for cold). The labels of collagen I and collagen III on the samples included in paraffin are performed after unmasking specific sites. The immunolabelings are carried out using a polyclonal rabbit antibody specific for collagen I (Rockland, Ref: 600-401-103-0.5), a polyclonal rabbit antibody specific for collagen III (Rockland , Ref: 600-401-105-0.5), followed by a secondary anti-rabbit antibody coupled to a fluorochrome (Invitrogen, Ref: A21206). For the labeling of hyaluronic acid, the cells are incubated in the presence of a specific biotinylated protein (Coger, Ref: 4007631A), then in the presence of streptavidin coupled to a fluorochrome (Invitrogen, Ref: 532354). Immunolabelling of pro-collagen, on samples included in OCT, is performed after a 10 minute fixation with cold acetone. Immunostaining is performed using a pro-collagen-specific rat polyclonal antibody (Millipore, Ref: MAB1912) followed by a secondary anti-rat antibody coupled to a fluorescent label (Invitrogen, A11006). The cells are then examined under an epifluorescence microscope (Nikon Eclipse E600 microscope). Results: The microscopic observations show a more intense dermal fluorescence of the skin samples treated with the small spelled extract according to Example 2. In the case of the immunolabeling of hyaluronic acid, there is also a increased fluorescence in the epidermis of the skin samples treated with the small spelled extract according to Example 2. Conclusions: The small spelled extract according to Example 2 stimulates the expression of collagen I, collagen III, pro-collagen in the dermis and stimulates the expression of hyaluronic acid in the human dermis and epidermis.

Example 3: Demonstration of the activating effect of the small spelled extract according to Example 2 on the expression of the collagen I messenger RNAs by real-time PCR The purpose of this study is to determine the influence of the small spelled extract according to Example 2 on the expression of messenger RNAs of collagen I in human fibroblasts. For this, the expression of collagen I was studied by real-time PCR. Protocol: Human fibroblasts in cultures are treated with the small spelled extract according to Example 2 at final concentrations of 0.5% and 1% for 24, 48 and 72 hours. The total RNAs are extracted using an extraction kit (QIAGEN), then reverse transcribed with a specific kit containing RNase inhibitors (Applied 2971711 -16- Biosystems). A real-time PCR is performed in a thermocycler using a TagMan Gene Expression Assay specific for Collagen I (Applied Biosystem, Hs99999901_sl) and an 18S-specific TagMan Gene Expression Assay used as an endogenous control (Applied Biosystem, Hs00164004_ml). The relative quantification of the messenger RNA expression of Collagen I is carried out by the comparative method of Ct (the Ct, threshold cycle, is the intersection between the amplification curve and the threshold line). Results / Conclusion: The small spelled extract according to Example 2 increases the level of expression of the messenger RNAs of Collagen I in human fibroblasts.

EXAMPLE 5 Demonstration of the Reversion of Celullary Senescence Induced in Fibroblasts by the Extract of Small Spelled spelled according to Example 2 The aim of this study is to determine the influence of small spelled extract according to US Pat. Example 2 on senescent fibroblasts, the senescence of which was induced by the extinction of the FOXO3a gene. These cells overexpress beta-galactosidase, in relation to their senescent state. FOXO3a is a Forkhead type transcription factor involved in cellular longevity. In the skin, the downregulation of FOXO3a accelerates the senescence of normal human fibroblasts (Hyun Kyoung K. et al., J. of Gerontol., 2005, 60A No. 1, pages 4-9). This property has been exploited to induce senescence in human fibroblasts by quenching the expression of FOXO3a by a specific interfering RNA (siRNA).

Protocol: Human fibroblasts in culture are made senescent by treatment with a FOXO3a-specific siRNA at a final concentration of 25 nM using the LipofectamineTM RNAiMAX transfection technique (Invitrogen, Ref: 13778-075). Untreated controls are performed. The fibroblasts are in parallel treated with the small spelled extract according to Example 2 at a final concentration of 1% for 48 hours. The cells are rinsed and fixed in fixing buffer (0.2% glutaraldehyde, 2% formaldehyde). The cells are then incubated at 37 ° C. without CO2 for 24 hours with a solution of X-Gal at 1 mg / ml in 40 mM citric acid / phosphate (pH 6), 5 mM K3FeCN6, 5 mM K4FeCN6 150 mM NaCl and 2 mM MgCl2. The cells are then examined under a white light microscope (Nikon Eclipse E600 microscope).

Results: Senescent cells with specific 13-galactosidase activity are stained blue. The cells treated with the FOXO3a-specific siRNA show an increase in the activity (3-galactosidase linked to the induction of senescence, resulting in an increase in the number of cells stained in blue. FOXO3a and the small spelled extract according to Example 1 at 1% show a decrease of the 13-galactosidase activity, resulting in a decrease in the number of cells stained blue.Conclusion: The extract of small spelled according to Example 2e decreases the senescence phenotype induced in human fibroblasts.

EXAMPLE 6 Preparation of Compositions 1 Anti-Aging Day Cream Trade Names INCI Names% by mass PHASE A MONTANOV 68 Cetearyl Alcohol (and) Cetearyl 5,00 Glucoside OJJOBA OIL Simmondsia Chinensis (Jojoba) Seed 3,00 Oil VASELINE OIL Paraffinum Liquidum (Ore Oil) 2.00 SQUALANE Squalane 3.00 CERAPHYL 368 Ethylhexyl palmitate 4.00 CERAPHYL 41 C12-C15 Alky1 Lactate 3.00 RAPTTHIX A-60 Sodium polyacrylate (and) 0.30 Hydrogenated Polydecene (and) Trideceth -6 PHASE B GLYCERIN Glycerin 5.00 ALLANTOIN Allantoin 0.10 DEMWERALIZED WATER Aqua (Water) qsp 100 PHASE C OPTIPHEN I Phenoxyethanol (and) caprylyl glycol i 1 PHASE D 2971711 -18- Trade names INCI names% by mass ACTIVE AGENT ACCORDING TO 5% EXAMPLE 1 PHASE E PERFUME Perfume (Fragrance) I qsp Procedure: Weigh the ingredients of the fat phase and heat to 70 ° C with stirring. Prepare phase B and heat it to 70 ° C. Emulsify phase A in phase B. Add phase C at 50 ° C with stirring. Below 30 ° C, add the active ingredient (Phase D). 5 Perfume and cool to room temperature.

2 - moisturizing cream W / O Trade names INCI names% by mass PHASE A ARLACEL P 135 PEG-30 Dipolyhydroxystearate 2.00 Isononanoate CERAPHYL 375 Isostearyl Neopentanoate 3.00 PANALANE L-14E Hydrogenated Polyisobutene 3.00 CERAPHYL ODS Octyldodecyl Stearate 9.00 CERAPHYL 368 Ethylhexyl Palmitate 3.00 PHASE B DEMINERALIZED WATER Aqua (Water) qs 100 ATLAS G-2330 Sorbeth-30 4.00 Magnesium Sulphate Sulfate 0.70 MAGNESIUM 7 H2O ACTIVE AGENT ACCORDING TO 2% EXAMPLE 2 PHASE C LIQUAPAR MET Phenoxyethanol (and) ) Methylparaben 0.50 (and) ethylparaben (and) propylparaben PHASE D PERFUME Perfume (Fragrance) qsp Procedure: Weigh phase A and heat to 75 ° C with stirring. Prepare phase B and heat it to 75 ° C. Emulsify phase B in phase A under strong rotor-stator agitation. 2971711 -19- Homogenize a few minutes. Cool suddenly with an ice-water bath with vigorous stirring. Add phase C at about 50 ° C and add the perfume (phase D) at 40 ° C. Continue cooling to room temperature.

Claims (19)

REVENDICATIONS1. Use of a small spelled extract (Triticum monococcum) in a cosmetic composition further comprising a physiologically adapted medium, as an activating active agent for the synthesis of the proteins of the extracellular matrix of the skin, to fight against the appearance cutaneous signs of aging caused by the degradation of the extracellular matrix. 2. Use of a small spelled extract according to claim 1 as an active agent stimulating the synthesis of type I and III collagens, fibronectin and hyaluronic acid by cutaneous cells. 3. Use of a small spelled extract according to claim 1 as an active agent to delay the onset or decrease senescence of skin cells. 4. Use of a small spelled extract according to one of the preceding claims, as an active agent for increasing the density of the dermis and the elasticity of the skin, and for delaying or reducing the loosening of facial features, loss of volume of the dermis, thinning of the skin, sluggishness, fine lines, deep wrinkles. 20 5. Use of a spelled extract according to one of the preceding claims, as an active agent to increase the ability of all layers of the skin to retain water and to delay or reduce dehydration of the skin related to aging. 25 6. Use according to one of the preceding claims, characterized in that said extract of small spelled contains between 1.5 and 3.5 g / 1 of peptide compounds. 7. Use according to one of claims 1 to 5, characterized in that said extract of small spelled contains between 1.5 and 3.5 g / 1 of peptide compounds with a molecular weight of less than 5 kDa. 8. Use according to one of the preceding claims, characterized in that said extract of small spelled is an aqueous extract diluted in one or more physiologically suitable solvents, such as water, glycerol, ethanol, propanediol, Butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. 2971711 -21- 9. Use according to one of the preceding claims, characterized in that said extract of small spelled is present at a concentration of between 0.0001% to 20% approximately, and preferably at a concentration between 0.05% and 5% approximately 5 relative to the total weight of the final cosmetic composition. 10. Use according to claim 9, characterized in that said composition further contains at least one other active agent. 10 11. Use according to claim 10, characterized in that the other active agent is selected from regenerating, anti-aging, anti-wrinkle, soothing, anti-radical, anti-glycation, moisturizing, antibacterial, antifungal, keratolytic, myorelaxant, desquamating, firming agents, agents stimulating the synthesis of dermal macromolecules or energy metabolism or microcirculation or nail growth or hair growth, agents modulating the differentiation or the pigmentation of the epidermis, the agents inhibiting metalloproteinases, or the screens or sunscreens. 12. Extract of small spelled (Triticum monococcum) obtained by the following process: a) Extraction by contacting crushed seeds or small spelled flour (Triticum monococcum) with an aqueous solution, with stirring, b) Successive hydrolyses with at least two proteases, c) Purification by at least one filtration step, d) Separation of low molecular weight compounds by ultrafiltration. 13. Small spelled extract according to claim 12 characterized in that the proteases are bromelain and Alcalase®. 14. Small spelled extract according to claim 13, characterized in that it comprises only peptide compounds with a molecular weight of less than 5 kDa. 15. Extract of small spelled according to claim 14 characterized in that it is diluted in one or more physiologically suitable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. 2971711 -22- 16. Cosmetic composition comprising a small spelled extract according to one of claims 12 to 15, at a concentration of between 0.0001% to 20% approximately, and preferably between 0.05% and 5% approximately relative to the total weight of the final cosmetic composition. 17. Cosmetic composition according to the preceding claim, characterized in that said composition also contains at least one other active agent. 10 18. Cosmetic composition according to the preceding claim, characterized in that the other active agent is selected from regenerating agents, anti-aging, anti-wrinkle, soothing, anti-radical, anti-glycation, moisturizing, antibacterial, antifungal, keratolytic, muscle relaxant , desquamating agents, firming agents, agents stimulating the synthesis of dermal macromolecules or energy metabolism or microcirculation or nail growth or hair growth, agents modulating the differentiation or the pigmentation of the epidermis, the agents inhibiting metalloproteinases , or screens or sunscreens. 19. Cosmetic treatment method for delaying and / or treating cutaneous signs of aging originating from the degradations of the extracellular matrix, characterized in that a composition comprising at least one extract is applied topically to the skin to be treated. small spelled spell, as defined in one of claims 16 to 18.