FR2915381A1 - Use of an active ingredient obtained from hydrolyzed soy, in a cosmetic composition or for preparing a pharmaceutical composition comprising the ingredient or composition to activate cytochrome and stimulate mitochondrion - Google Patents

Abstract

Use of an effective quantity of at least an active ingredient (I) obtained from hydrolyzed soy ( Glycine max L.), in a cosmetic composition or for preparing a pharmaceutical composition comprising (I) or composition to activate cytochrome and stimulate mitochondrion, is claimed. ACTIVITY : Dermatological; Muscular-Gen; Neuroprotective; Cardiant; Antidiabetic; Nootropic. MECHANISM OF ACTION : Cytochrome activator. Tests details are described but no results given.

Description

The present invention is in the field of cosmetics and

  pharmaceutical, and more particularly in the field of dermatology. The present invention relates to the use, in a cosmetic composition or for the preparation of a pharmaceutical composition, of an effective amount of active ingredient derived from soybean (Glycine Max L.); the active ingredient, or the composition containing it, being intended to activate cytochrome c. Preferably, the active ingredient is derived from the hydrolysis of soy protein and contains mainly polypeptides or peptides. It can be used alone or in combination with at least one other active ingredient. The invention also relates to the use of a cosmetic composition intended to stimulate the functions of mitochondria and to increase the cellular energy level. The invention also relates to a cosmetic treatment method intended to protect the skin and the integuments from external aggressions and to fight against skin aging. The active ingredient, activator of cytochrome c, can also be used to prepare pharmaceutical compositions intended to prevent or fight against pathologies related to mitochondrial dysfunctions; for example, certain neuromuscular or cardiac degenerations, type II diabetes or certain pathologies of aging.

  The term dander according to the invention encompasses all the keratinous appendages present on the surface of the body, in particular the hairs, the eyelashes, the eyebrows, the nails and the hair.

  The skin is a vital organ that covers the entire surface of the body and provides protective, sensitive, immune, metabolic or thermoregulatory functions. The skin, like other organs, is subject to aging. However, one of the major mechanisms involved in aging processes is the accumulation of oxidative damage in essential molecules such as membrane lipids, proteins, DNA and especially mitochondrial DNA (mtDNA).

  Oxidative damage is caused by free radicals, chemically unstable and highly reactive species generated by intracellular metabolism or external aggression. These external aggressions include: UV radiation, toxins, air pollutants, food oxidants. In the skin, premature aging occurs in areas exposed to radiation, characterized by phenomena of alterations of macromolecules (lipid peroxidation, carbonylation of proteins) particularly affecting elastin, collagen or fibronectin. . It has also been possible to show a progressive decline in mitochondrial functions with age, probably related to the accumulation of mutations on mtDNA (K. Singh, Ann N.Y. Acad Sci 1019, 2004). One of the important consequences of the accumulation of oxidative damage is a reduction in the ability of the cell to produce ATP (Porteous et al., Eur J Biochem 1998, 257 (1): 192-201). Thus, the phenomenon of cellular aging is related to the oxidative damage that the cell undergoes but also to the process of producing energy necessary for the cell to survive. The body has defense mechanisms capable of trapping or transforming free radicals (enzymes, glutathione, vitamins A and E, coenzyme Q10, etc.). However, these systems of antioxidant defenses often prove insufficient in the face of the numerous stresses and external aggressions to which the organisms and the skin in particular are subjected. In this context, the particular properties of cytochrome c appear to be of particular interest: Cytochrome c is a small, 15 kDa soluble protein that plays a critical role in mitochondrial function and cell survival. Cytochrome c is a highly conserved molecule in most eukaryotes; it is found in the mitochondria of plants, animals and many unicellular organisms. Cytochrome c has a protein structure organized around a porphyrin, consisting of four pyrrol nuclei, themselves linked to an iron atom. The main role of cytochrome c is to ensure the transfer of electrons, thanks to the change of valence of the iron atom. Cytochrome c, which is soluble, transports electrons from complex III (coenzyme QH2-cytochrome c reductase) to complex IV (cytochrome oxidase). The electrons, which are the substrate for cytochrome oxidase, are then transferred by the enzyme to oxygen. The search for compounds that can stimulate mitochondria and increase the cellular energetic level in order to prevent or fight against the signs of skin aging, or the damage caused by external aggressions, such as UV-radiation, radiations, or exposures to toxins or pollutants, is a major concern of medical research and cosmetics. In this regard, solutions have been proposed such as the supply of substances involved in energy metabolism, and more particularly Krebs cycle intermediates or cofactors such as fumarate, L-malate, acetyl CoA (WO 02064129) or the treatment of the skin with substances capable of reducing free radicals, such as vitamin C (US 2004/0086526) or L-ergothioneine (WO 9836748). But to the knowledge of the applicant, no cosmetic or pharmaceutical composition comprising peptide compounds derived from soy, capable of activating cytochrome c has yet been described.

  The main objective of the present invention is the use of an active ingredient derived from the hydrolysis of soybeans, capable of protecting the skin against external aggressions and of combating cutaneous aging. Said active ingredient may be used alone or in combination with at least one other active ingredient. The inventors have in fact demonstrated a therapeutic activity, and more particularly a dermatological and cosmetic activity, of an active principle of a peptide nature resulting from the hydrolysis of soybean, capable of activating cytochrome c. In particular, it has been demonstrated that these polypeptides or peptides, which constitute the active principle, when applied to the skin, significantly stimulate the mitochondrial functions. This has, among other things, been demonstrated by an increase in the expression of cytochrome c, an increase in cytochrome oxidase activity and an increase in ATP synthesis. This new stimulating active principle of mitochondria, and more generally capable of protecting the skin from external aggressions, thus opens up new therapeutic and cosmetic perspectives. The expression "active principle" capable of protecting the skin against external aggressions is understood to mean any substance of plant origin, and more particularly derived from soy (Glycine Max L.), capable of exhibiting protective properties or of decreasing apoptosis in cells. or tissues subjected to stress of physicochemical or environmental origin. The term "active ingredient capable of activating cytochrome c" means any substance of plant origin, and more particularly derived from soy (Glycine Max L.), capable of increasing the expression of cytochrome c, either by activation protein synthesis (by direct or indirect modulation of cytochrome c gene expression), either by increasing the biological activity of cytochrome c, or by other biological processes such as the stabilization of the cytochrome c protein or the stabilization of messenger RNA transcripts. By active ingredient capable of stimulating the functions of the mitochondria is meant any substance of plant origin, and more particularly derived from soy (Glycine Max L.), capable of increasing the expression or the activity of the main active molecules present in the mitochondria, and more generally capable of increasing the main energy functions of the mitochondria; that is, the chain of oxidative phosphorylation and ATP synthesis. Preferably, the active ingredient capable of activating cytochrome c according to the invention is an extract of a peptide nature and comes from the hydrolysis of soy proteins (Glycine Max L.). By peptide nature is meant a mixture of compounds predominantly represented by peptides or polypeptides. The term peptide denotes a sequence of two or more amino acids linked together by peptide bonds or by modified peptide bonds; the term polypeptide designating a peptide of larger size. By biologically active expression is meant that which has an activity in vivo or in vitro characteristic of the activity of the active ingredient according to the invention. The term "hydrolyzate or derived from hydrolysis" refers to any substance or mixture of substances, or isolated preparation, obtained after hydrolysis of plant material.

  The active ingredient according to the invention can be obtained by extraction of proteins of plant origin, followed by controlled hydrolysis which liberates biologically active peptide fragments. The 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 either extract the relevant proteins first and then hydrolyze them, or to hydrolyze first on a crude extract and purify the peptide fragments. then. It is also possible to use certain hydrolysed extracts without purifying the peptide fragments corresponding to the biologically active peptides according to the invention, but nevertheless ensuring the presence of said fragments by appropriate analytical means. To carry out the extraction, one can use the whole plant, or a specific part of the plant (leaf, grain, etc.). More particularly according to the invention, one of the many plants of the Fabaceae family, of the genus Glycine, soybean or soybean meal is used, and preferentially Glycine Max species. L. According to the invention, the plant material used will be the grain and preferentially the grain freed of its envelope by a dehulling step. In a first step, the plant is milled using a plant mill. The powder thus obtained may subsequently be "delipidated" with the aid of a conventional organic solvent (for example an alcohol, hexane or acetone). The proteins of the plant are then extracted according to the conventional method (Osborne, 1924) modified; the plant mash is suspended in an alkaline solution containing an insoluble polyvinylpolypyrrolidone (PVP) adsorbent product (0.01-20%); in fact, it has been observed that the hydrolysis and subsequent purification operations are facilitated by this means. The concentration of phenolic-type substances interacting with the proteins is thus reduced. The soluble fraction is collected after centrifugation and filtration steps, this crude solution then constituting a first form of the extract containing proteins, carbohydrates and possibly lipids. The proteins are then precipitated by varying the ionic strength by acidifying the medium, which eliminates soluble components and nucleic acids. The precipitate is then washed with an organic solvent such as, for example, ethanol or methanol and then the solvent is evaporated by drying in vacuo. The protein-rich precipitate is dissolved in water or other solvent and is then a more purified form of the hydrolyzate. The extraction can also be carried out in neutral or acidic medium always in the presence of polyvinylpolypyrrolidone. After a filtration step, the precipitation step is then carried out using a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate) or an organic solvent (alcohol, acetone) . The precipitate obtained can be separated from the precipitating agents by dialysis after redissolving in water or another solvent. The isolated protein fraction according to the invention is then hydrolyzed under mild conditions to generate polypeptides and soluble peptides. 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 carried out chemically and / or advantageously by proteolytic enzymes. We can then mention the use of endoproteases of plant origin (papain, bromelain, ficin) and microorganisms (Aspergillus, Rhizopus, Bacillus, etc.). For the same reasons as above, during this controlled hydrolysis step, an amount of polyvinylpolypyrrolidone is added to the reaction medium. After filtration, the solution obtained constitutes the active hydrolyzate. The active hydrolyzate can be further purified in order to select the molecular weights and the nature of the peptides generated. The fractionation can advantageously be carried out by ultrafiltration and / or by a chromatographic type method. Any of the more or less purified forms of the hydrolyzate is then solubilized in water or in any mixture containing water, and then sterilized by ultrafiltration. The vegetable hydrolyzate obtained according to the invention is analyzed qualitatively and quantitatively for its physicochemical characteristics and its content of protein and peptide compounds. Peptide-like compounds are understood to mean protein fragments, peptides and 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. Thus, according to an advantageous embodiment of the invention, the active plant hydrolyzate has a pH of between 4 and 7, and preferably between 5 and 6, a solids content of between 1 and 8 g / l, and preferred manner between 2 and 5 g / 1, its content of peptide compounds is between 0.1 and 5 g / l, and preferably between 0.5 and 2 g / 1 and its sugar content is 0.5 to 2.5 g / l.

  According to an advantageous embodiment of the invention, the active principle according to the invention is first solubilized in one or more cosmetically or pharmaceutically acceptable solvents, conventionally used by those skilled in the art, such as water, glycerol ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols, petroleum jelly, a vegetable oil or any mixture of these solvents. According to yet another advantageous embodiment of the invention, the active ingredient according to the invention is first solubilized in a cosmetic or pharmaceutical vector such as liposomes or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites, and more generally solubilized in, or attached to, any cosmetically or pharmaceutically acceptable carrier. The composition that may be used according to the invention may in particular consist of a composition for hair care, and in particular a shampoo, a conditioner, a setting lotion, a treatment lotion, a styling cream or gel, a restructuring lotion. for the hair, a mask, etc. The cosmetic composition according to the invention can be used in particular in treatments using an application that is followed or not followed by rinsing, or in the form of shampoo. It can also be in the form of dye or mascara to be applied with a brush or a comb, in particular on eyelashes, eyebrows or hair. It is understood that the active ingredient according to the invention may be used alone or in combination with at least one other active ingredient, in a cosmetic composition or for the preparation of a pharmaceutical and / or dermatological composition. The compositions according to the invention may be applied by any appropriate route, in particular oral, parenteral or external topical, and their formulation will be adapted by those skilled in the art, in particular for cosmetic or dermatological compositions. Advantageously, the compositions according to the invention are intended for topical administration to the skin. These compositions must therefore contain a cosmetically and / or dermatologically acceptable medium, that is to say compatible with the skin and superficial body growths, and cover all cosmetic or dermatological forms. These compositions may especially be in the form of creams, oil-in-water or water-in-oil emulsions or multiple emulsions, solutions, suspensions, gels, milks, lotions, sticks or even powders, suitable for application on the skin. skin, lips and / or integuments. These compositions comprise the excipients necessary for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes. Advantageously, the compositions that can be used according to the invention also contain various active principles intended, in particular, for the prevention and / or treatment of disorders associated with photoaging. Thus, the composition according to the invention can associate, with the active principle according to the invention, other active ingredients promoting its action. For example, active ingredients having an anti-radical or antioxidant action selected from vitamin C, vitamin E, coenzyme Q10 and polyphenolic extracts of plants may be added. The composition according to the invention can also associate, with the active principle according to the invention, active principles stimulating the synthesis of dermal macromolecules (laminin, fibronectin, collagen), for example the collagen peptide sold under the name Collaxyl by the company Vincience . The composition according to the invention may also associate, with the active principle according to the invention, active principles stimulating the energy metabolism, such as the active ingredient sold under the name GP4G by the company Vincience. According to another aspect, the composition according to the invention may be a solar composition, that is to say a composition helping to protect against solar radiation. Thus, it is possible to advantageously add to the composition according to the invention, active agents that help to protect the sun such as, for example, sunscreens. It is obvious that the invention is directed to mammals in general and more particularly to humans. The effective amount of active ingredient corresponds to the amount necessary to obtain the desired result, namely: activate the cytochrome c, stimulate the mitochondria and increase the cellular energy level, and more generally, protect the skin and the integuments of the aggressions external and fight against skin aging According to an advantageous embodiment of the invention, the active ingredient is present in the compositions of the invention at a concentration of between 0.0001% and 20% approximately, and preferably at a concentration of between 0.05% and 5% relative to the total weight of the final composition. These compositions may especially be in the form of an aqueous solution, hydroalcoholic or oily; an oil-in-water, water-in-oil emulsion or multiple emulsions; they 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, gel, paste or paste. a foam. They can also be in solid form, as a stick or be applied to the skin in aerosol form. They can be used as a care product and / or as a make-up product for the skin. These compositions additionally comprise any additive commonly used in the intended field of application as well as the adjuvants necessary for their formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens. , self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc. In all cases, those skilled in the art will ensure that these adjuvants and their proportions are chosen so as not to adversely affect the desirable properties of the composition according to the invention. These adjuvants may, for example, correspond to 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, relative to the total weight of the composition.

  By virtue of its particular activities, the active principle according to the invention may advantageously be used in a cosmetic composition or for the preparation of a pharmaceutical composition. In particular, the active ingredient according to the invention may advantageously be used in a cosmetic composition intended to preventatively and / or curatively fight against the manifestations of skin aging and, more specifically, in order to fight against and / or prevent photo-induced aging (photo-aging). Skin manifestations of aging means any changes in the external appearance of the skin and skin appendages due to aging, such as, for example, wrinkles and fine lines, wilted skin, soft skin, thinned skin, lack of elasticity and / or tone of the skin, dull and dull skin or skin pigmentation spots, discoloration of the hair or spots on the nails, but also any internal changes in the skin that do not This does not necessarily result in a modified external appearance, such as, for example, any internal skin degradation resulting from exposure to ultraviolet (UV) radiation. The active ingredient according to the invention, or the composition containing it, will make it possible, in particular, to combat the loss of elasticity and firmness of the skin. The active ingredient according to the invention makes it possible to protect the skin and integuments against all types of external aggressions. The use of the active ingredient, or a composition containing it, will allow the skin and integuments to be protected and to better withstand environmental stresses. By the term external aggression, we mean the aggressions that the environment can produce. By way of example, mention may be made of aggressions such as pollution, UV, or irritating products such as surfactants, preservatives or perfumes. Pollution is understood to mean both external pollution, eg due to diesel particles, ozone or heavy metals, as well as internal pollution which may be due to solvent emissions from paints, adhesives or paper. (such as toluene, styrene, xylene or benzaldehyde), or even cigarette smoke. The active ingredient according to the invention can be advantageously used in a cosmetic composition or for the preparation of a pharmaceutical composition, as a photoprotective agent and, more particularly, as a so-called secondary photory protective agent. In fact, the primary photoprotective agents are distinguished from the secondary photoprotective agents. Primary photoprotective agents are substances that exert physical power: they are able to absorb UV radiation and release it as heat to protect the skin. Secondary photoprotective agents are substances that usually have a biological effect; these are, for example, agents capable of limiting the damage to DNA and membranes caused by the penetration of UV radiation into the skin.

  The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of an effective amount of

  active ingredient according to the invention, the active ingredient or the composition containing it, being intended to increase the synthesis of intracellular ATP of the cells of the skin. The subject of the invention is also the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of an effective quantity of active principle according to the invention, the active principle, or the composition containing it, being intended to to prevent damage to the skin caused by exposure to the sun or exposure to ionizing radiation during radiation therapy. The subject of the invention is also the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of an effective quantity of active principle according to the invention, the active principle, or the composition containing it, being intended to to stimulate the mitochondria, in particular on the areas of the body exposed to UV radiation. The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of an effective amount of active ingredient as described above, the active ingredient, or the composition containing it, being intended to to protect the skin from damage caused by free radicals. The invention also consists in a pharmaceutical composition characterized in that the active principle according to the invention is formulated to attenuate a pathology related to mitochondrial dysfunctions, for example certain neuromuscular or cardiac degenerations, type II diabetes, certain pathologies of aging. The invention also consists of a cosmetic treatment method intended to stimulate the defenses and to protect the skin and integuments from external aggressions and to fight against skin aging, characterized by the application on the skin or integuments to be treated of a composition containing an effective amount of active ingredient according to the invention.

  Particular embodiments of this cosmetic treatment method also result from the foregoing description. Other advantages and characteristics of the invention will appear better on reading the examples given by way of illustration and not limitation. EXAMPLE 1 Preparation of Active Ingredient from Desofa Cake (Glycine Max L.) Cakes are the solid residues obtained after extraction of the oil from the soya beans. They represent 50 to 75% of the seed mass. The active ingredient is obtained from plants of the species Glycine max L. Of course the extract can be prepared from plants of at least one of the many varieties and species belonging to the genus Glycine. In a first step, 1 kg of soybean meal is milled in a cereal mill. The flour obtained is delipidated by the action of an organic solvent, hexane. After filtration and drying under vacuum, the powder obtained is suspended in an aqueous alkaline solution (1/10 dilution) pH 10 containing 1% polyvinylpolypyrrolidone (Polyclar V ISP). This mixture is stirred for a time long enough to allow the solubilization of the soluble fractions. The extraction temperature is variable (between 4 and 80 C); Preferentially the operation will be performed cold. After this extraction phase, the medium is clarified by centrifugation and then filtered through a plate filter. This filtrate, which contains the soluble fractions of soybeans, is then subjected to precipitation of the proteins by varying the ionic strength in a neutral or acid medium, which makes it possible to eliminate the soluble carbohydrate components, the lipids and the nucleic acids. at pH 3.5. The supernatant is removed and the precipitate is then washed with a solvent such as, for example, ethanol or methanol and the solvent is evaporated by drying under vacuum. At this stage, about 50 grams of light yellow powder of crude protein extract are obtained containing: Proteins: 75% Carbohydrates: 20% Lipids: 5%

  The protein-rich precipitate is dissolved in water or another solvent. The crude protein extract is then subjected to a series of controlled and selective hydrolyses consisting of chemical and enzymatic hydrolyses in the presence of 0.5% PVPP (Polyclar V) and cysteine endopeptidases (papain, ficin). After reaction, the hydrolyzate is filtered on a plate and then on a sterilizing cartridge (0.21 μm). A light-colored hydrolyzate, titrating with 15 to 30 g / l of dry extract, is then obtained, which is then diluted so that the concentration of peptidic compounds determined by the method of Lowry is included between 0.1 and 5 g / l and preferably between 0.5 and 2 g / l. The physicochemical analysis of the vegetable hydrolyzate, which constitutes the active principle, shows that its pH is between 4 and 7, and preferably between 5 and 6, the solids content is 1 to 8 g / l and preferably between 2 and 5 g / 1, its content of peptide compounds is between 0.1 and 5 g / 1, and preferably between 0.5 to 2 g / 1 and its sugar content between 0.5 at 2.5 g / 1. EXAMPLE 2 Preparation of Active Ingredient from Sofa (Glycine Max L.) A variant of the protocol of Example 1 consists in carrying out the same sequence of mild and selective enzymatic hydrolyses but in the presence of 0.5% of PVPP. A light-colored hydrolyzate titrating with 15 to 30 g / l of dry extract is then obtained after sterilizing filtration. The solution is then ultrafiltered on a Millipore Helicon filtration cartridge (cut-off point, 1 kDa). The high molecular weights contained in the retentate are removed, the filtrate is preserved. The concentration of compounds of peptide nature is determined by the method of Lowry, between 0.1 and 5 g / l and preferably between 0.5 and 2 g / l. The physicochemical analysis of the vegetable hydrolyzate, which constitutes the active principle, shows that its pH is between 4 and 7, and preferably between 5 and 6, the solids content is between 1 and 8 g / l. . and preferably between 2 and 5 g / l, its content of peptide compounds is between 0.1 and 5g / 1 and preferably between 0.5 to 2 g / 1 and its sugar content between 0.5 to 2.5 g / l.

  Another variant consists in carrying out a purification of the active principle, obtained according to example 1 or 2, by ion exchange chromatography, on a TSK gel column (TosoHaas) with a phosphate buffer pH 7. EXAMPLE 3 Evidence of the stimulating effect of the active principle according to Example 1 on the synthesis of intracellular ATP The purpose of this study is to determine the influence of the active principle according to Example 1 on the synthesis of ATP, produced by the mitochondria. 2915381 -14- Protocol: This study is carried out using an ATP Bioluminescence Kit Assay Kit HS II (Roche Applied Science). Dermal fibroblasts are treated with a 1% solution, of active principle according to Example 1, for a period ranging from 1 to 3 hours. At the end of the incubation, the wells are rinsed with 2 ml of cold PBS before adding 250 μl of lysis buffer provided by the kit. The cells of each well are then scraped and then harvested in 14 ml tubes. Each well is rinsed with 2 x 500 l PBS cold and all is harvested again in the respective tubes. From these samples, a dilution is made at 1 / 120001eme in cold PBS before each reading. ATP assay is performed on these samples: 50 L of this dilution are deposited in a luma-cuvette and 50 tL of luminol are added. After 10 seconds, the reading of the luminescence is triggered. The values are standardized with respect to the amount of protein for each sample. The measurements are made using a device: the Biocounter M2010A LUMAC / 3M. Results: The ATP assays show that there is a significant increase in the amount of intracellular ATP after 1 hour and 3 hours, in cells treated with the active ingredient according to Example 1, compared to non-intact cells. processed. Conclusion: The active ingredient according to Example 1 substantially increases the energy level of cutaneous cells such as fibroblasts, and more generally stimulates mitochondrial functions. EXAMPLE 4 Demonstration of the Activating Effect of the Active Principle According to Example 1 on the Expression of Cytochrome c The aim of this study is to determine the influence of the active principle according to Example 1 on the expression cytochrome c. For this, the quantity of cytochrome c was evaluated by the technique of immuno-transfer (or western blot). Protocol: Human dermal fibroblasts are treated with a 1% solution of the active ingredient according to Example 1 for 72 hours. The cells are then lysed and homogenized by sonication, and then centrifuged for 10 minutes at 1000 g. The samples, standardized for their protein concentration (BCA kit assay, Pierce), are subjected to 4-12% Bis-TRIS gel electrophoresis (InvitroGen). After electro-transfer, the membranes are incubated overnight with an anti-cytochrome c antibody, diluted 1/500 (Mouse monoclonal anti cytochrome c, TEBU). A secondary antibody coupled to peroxidase and diluted 1/5000 is then used (conjugated peroxidase F (ab ') 2, Goat antimouse, Immunotech). The chemiluminescent signal is then quantified using the Supersignal Kit West Femto Trial kit and read in a reading chamber (Multilage Light Cabinet, Alpha Immunotech Corporation). Results: A marked increase in the expression of cytochrome c in the fibroblasts treated with the active principle according to Example 1 is observed. Conclusions: The active principle according to Example 1 strongly stimulates the expression of cytochrome c in cells cutaneous, and more generally the mitochondrial functions. EXAMPLE 5 Demonstration of the Activating Effect of the Active Principle According to Example 1 on the Enzyme Activity of Cytochrome Oxidase The aim of this study is to determine the influence of the active ingredient according to Example 1 on the mitochondrial activity. For this, the total enzymatic activity (mitochondrial) of cytochrome oxidase was measured. Protocol: Normal human fibroblasts are treated with a 1% solution of the active ingredient according to Example 1 for 3 hours and 24 hours. The cells are harvested, rinsed and then lysed by sonication. A first centrifugation is carried out to remove the main cellular debris and the supernatant is then centrifuged again at 10,000 g for 10 min. The enzymatic activity is assayed in the supernatant and / or in the 2nd pellet, by a biochemical method, using Cytocox 1 kit (Sigma) then standardized for the protein content (assayed by the BCA kit). Results: The assays show that there is a very significant increase in the enzymatic activity of cytochrome oxidase after 3 hours and 24 hours of application of the active principle according to Example 1, in comparison with non-enzymatic cells. The active ingredient according to Example 1 very strongly stimulates the enzymatic activity of cytochrome oxidase, and more generally the mitochondrial activity, in cutaneous cells. Protective effect of the active principle according to Example 1 on the mitochondrial membrane potential The purpose of this study is to determine the protective effect of the active ingredient according to Example 1, with respect to the mitochondria of dermal fibroblasts subjected to a oxidative stress, caused by hydrogen peroxide (H2O2) or UVB irradiation. A marker of the membrane potential of mitochondria (JC-1) is used for this purpose. JC-1 is a marker that emits a different fluorescence depending on the level of polarization of the mitochondrial membrane. Protocol: The human dermal fibroblasts are treated with a 1% solution of active principle according to Example 1, for 96 hours, then subjected to oxidative stress, caused by H2O2 at 2 mM for 30 minutes, or at room temperature. irradiation with UVB at 50 mJ / cm 2. Controls not treated with the peptide or with H2O2 or non-irradiated are carried out under the same conditions. At the end of the experiment, the cells are washed, fixed and labeled with a solution of JC-1 (Molecular Probes) at 0.2 g / ml, in order to reveal the membrane potential of the mitochondria. Results: In the fibroblasts treated with the active principle according to Example 1, the mitochondria exhibit a red fluorescence (JC-1 aggregated), a sign of a high membrane potential, greater than in the control cells. Irradiated or oxidatively stressed fibroblasts have mitochondria that fluoresce little in the red, and mainly in green (monomeric JC-1), a sign of altered mitochondrial membrane potential. In these latter conditions, the application of the active ingredient according to Example 1 makes it possible to observe a more pronounced red fluorescence. Conclusions: The application of the active ingredient according to Example 1 causes an increase in the membrane potential of the mitochondria. On the other hand, the active ingredient according to Example 1 effectively protects the mitochondria of cutaneous cells subjected to oxidative stress or irradiation with UVB.

  EXAMPLE 7 Preparation of Compositions 1 - Sunscreen Cream: Trade Names INCI Names% by mass PHASE A Demineralised water ~ Aqua (Water) qsp -17- Trade names INCI names% by weight Pemulen TRI Acrylates / C10-30 Alkyl Acrylate 0.40 Crosspolymer Glycerin Glycerin 3.00 Nipastat Sodium Sodium Methylparaben (and) Sodium 0.15 Ethylparaben (and) Sodium Butyl paraben (and) Sodium Propylparaben (and) Sodium Isobutylparaben PHASE B Parsol MCX Ethylhexyl Methoxycinnamate 7.50 Eusolex 4360 Benzophenone-3 3.00 Parsol 1789 Butyl Methoxydibenzoylmethane 2.00 Myritol 318 Caprylic / Capric Triglyceride 4.00 Emulgade SEV Hydrogenated Palm Glycerides (and) 5.00 Ceteareth-20 (and) Ceteareth-12 (and) Cetearyl Alcohol Propylparaben Propylparaben 0.15 Nacol 16-98 Cetyl Alcohol 1.00 PHASE C TEA Triethanolamine 0.20 PHASE D Active principle of Example 1 3 Fragrance Perfume (Fragrance) qsp Colorant qsp The constituents of phase A and phase B are heated separately between 70 C and 75 C . Phase B is emulsified in phase A with stirring. Phase C is added at 45 ° C., increasing stirring. Phase D is then added when the temperature is below 40 C. Cooling is continued to 25 C with vigorous stirring.

  2 ûLait après-soleil: Trade names INCI names% by mass PHASE A 2915381 -18- Trade names INCI names% by weight Montanov L C14-22 Alcohols (and) C12-20 3.00 Alkyl Glucoside Waglinol 2559 Cetearyl Isononanoate 4.00 Tegosoft TN C12-15 Alkyl Benzoate 3.00 Apricot Kernel Oil Prunus Anneniaca (Apricot) Kernel 2.00 Oil Avocado Oil Persea Gratissima (Avocado) Oil 1.00 Abil 350 Dimethicone 1.00 PHASE B Demineralized Water Aqua (Water ) 1 qsp PHASE C Simulgel EG Sodium Acrylate / Acryloyldimethyl 0.4 Taurate Copolymer (and) Isohexadecane (and) Polysorbate 80 Copolymer (and) Polysorbate 80 PHASE D Phenonip Phenoxyethanol (and) 0.30 Methylparaben (and) Ethylparaben (and) Butylparaben (and) Propylparaben (and) Isobutylparaben Ethylparaben and Propylparaben and Buthylparaben! Germ 115 Imidazolidinyl Urea 0.20 PHASE E Active ingredient of Example 1 1 0.1 Prepare phase A with stirring. Incorporate the xanthan gum gradually, with deflocculating stirring. Phases C and D will be incorporated once the gel is complete. The phase E, prepared previously until perfect dissolution of the DHA, will then be added. Adjust the pH if necessary to 4 - 4.5. Color and perfume.

  3 - Anti-aging Cream: Names INCI Names% commercial mass A Phase Montanov 68 Cetearyl Alcohol (and) Cetearyl Glucoside 6.00 Squalane Squalane 3.00 Cetiol SB 45 Butyrospermum Parkii (Shea Butter) 2.00 Waglinol 250 Cetearyl Ethylhexanoate 3.00 Amerchol L- 101 Ore Oil (and) Lanolin Alcohol 2.00 Abil 350 Dimethicone 1.50 BHT BHT 0.01 Coenzyme Q10 Ubiquinone 0.10 Phase B Avocado Oil Persea Gratissima (Avocado) Oil 1.25 Phenonip Phenoxyethanol (and Methylparaben (and) Ethylparaben 0.75 (and) Butylparaben (and) Propylparaben (and) Isobutylparaben Phase C Demineralized Water Aqua (Water) qsp Butylene Glycol Butylene Glycol 2.00 Glucam El0 Methyl Gluceth-10 1.00 Allantoin Allantoin 0, Carbopol Ultrez 10 Carbomer 0.20 Phase D TEA Triethanolamine 0.18 -20- Names INCI Names% Commercial Mass E Phase Active Principle 0.5 Example 1 GP4G Water (and) Artemia Extract 1.50 Collaxyl Water (and) ) Butylene Glycol (and) Hexapeptide-9 3.00 Phase F Perfume Perfume (Fragrance) qsp Colorant q sp Prepare and melt phase A at 65-70 C. Heat phase C at 65-70 C. Phase B is added to phase A just before emulsifying A in B. At about 45 C, the carbomer is neutralized by addition of phase D. Phase E is then added with gentle stirring and cooling is continued to 25 C. Phase F is then added if desired.

  4 - Protective Day Cream: Trade Names INCI Names% by mass Phase A Emulium Delta Cetyl alcohol (and) Glyceryl Stearate (and) PEG- 4.00 75 Stearate (and) Ceteth-20 (and) Steareth-20 Lanette O Cetearyl Alcohol 1 , 50 DC 200 Fluid / 100cms Dimethicone 1.00 DUB 810C Coco Caprylate / Caprate 1.00 DPPG Propylene Glycol Dipelargonate 3.00 DUB DPHCC Dipentaerythrityl Hexacaprylate / Hexacaprate 1.50 Cegesoft PS6 Vegetable Oil 1.00 2915381 -21- Vitamin E Tocopherol 0.30 Phenonip Phenoxyethanol (and) Methylparaben (and) 0.70 Ethylparaben (and) Butylparaben (and) Propylparaben (and) Isobutylparaben Phase B Demineralized Water Aqua qs 100 Glycerin Glycerin 2.00 Carbopol EDT 2020 Acrylates / C10-30Alkyl Acrylate Crosspolymer 0.15 Keltrol BT Xanthan Gum 0.30 Phase C Sodium Hydroxide (Sodium Hydroxide Sodium 0.30 10%) Phase D Demineralized Water Aqua 5.00 Stay-C 50 Sodium Ascorbyl Phosphate 0.50 Phase E Butylene Glycol Butylene Glycol 2.00 Dekaben CP Chlorphenesin 0.20 Phase F GP4G Water (and) Artemia Extract 1.00 Active ingredient of Example 1 Prepare phase A and heat to 75 ° C. with stirring. Prepare phase B by dispersing the carbopol, then the xanthan gum with stirring. Let rest. Heat to 75 C. At temperature, emulsify A in B with rotor-stator stirring. Neutralize with phase C with rapid stirring. After cooling to 40 ° C., add phase D and then phase E. Cooling is continued with gentle stirring and phase F added.

Claims (14)

  Use of an effective amount of at least one active principle, derived from the hydrolysis of soybean (Glycine Max L.), in a cosmetic composition or for the preparation of a pharmaceutical composition, characterized in that the active ingredient, or the composition containing it, is intended to activate cytochrome c and stimulate mitochondria.   2. Use according to claim 1, characterized in that the active principle is of peptide nature and comes from the hydrolysis of soy proteins (Glycine Max L.).   3. Use according to any one of the preceding claims, characterized in that the active ingredient contains at least 0.1 and 5 g / 1, and preferably at least 0.5 to 2 g / 1 of peptide compounds.   4. Use according to any one of the preceding claims, characterized in that the active ingredient is used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing 0.05 % to 5% of the total weight of the composition.   5. Use according to any one of the preceding claims, characterized in that the active ingredient is first solubilized in one or more cosmetically or pharmaceutically acceptable solvents, such as water, glycerol, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols, petroleum jelly, a vegetable oil or any mixture of these solvents.   6. Use according to any one of the preceding claims, characterized in that the composition is in a form suitable for topical application comprising a cosmetically or dermatologically acceptable medium. 2915381 -23-   7. Use according to any one of the preceding claims, characterized in that the composition additionally contains at least one antioxidant active principle and / or at least one active principle stimulating the synthesis of the extracellular matrix, and / or at least one an active ingredient stimulating energy cellular metabolism.   8. Use of an effective amount of active ingredient, as defined in any one of claims 1 to 3, in a cosmetic composition or for the preparation of a pharmaceutical composition for protecting the skin and integuments against all types of external aggression.   9. Use of an effective amount of active ingredient, as defined in any one of claims 1 to 3, in a cosmetic composition or for the preparation of a pharmaceutical composition for preventing or treating damage to the skin. the skin and to the integuments by UV radiation.   10. Use of an effective amount of active ingredient as defined in any one of claims 1 to 3 in a cosmetic composition or for the preparation of a pharmaceutical composition for stimulating and / or protecting mitochondria.   11. Use of an effective amount of active ingredient as defined in any one of claims 1 to 3 in a cosmetic composition or for the preparation of a pharmaceutical composition for increasing the synthesis of intracellular ATP of skin cells.   12. Use of an effective amount of active ingredient, as defined in any one of claims 1 to 3, in a cosmetic composition or for the preparation of a pharmaceutical composition for preventing or treating cutaneous signs of the skin. aging and / or photo-aging. 5 2915381 -24-   13. Use of an effective amount of active ingredient, as defined in any one of claims 1 to 3, for preparing pharmaceutical compositions for preventing or combating mitochondrial dysfunction.   14. Cosmetic treatment process intended to stimulate the defenses and to protect the skin and the integuments of the external aggressions, characterized in that one applies topically on the skin or the integuments to treat a composition as defined according to any one Claims 1 to 7 10