FR3006588A1 - PEPTIDE SOYBEAN EXTRACT AND ITS COSMETIC USE TO STRENGTHEN THE STRUCTURE OF THE HAIR - Google Patents

Abstract

The present invention is in the cosmetic field. The invention relates to the cosmetic use of a peptide extract of soy (Glycine Max L.), derived from the hydrolysis of soybeans, as an agent for strengthening the structure of the hair. The invention also relates to the cosmetic use of a soy peptide extract (Glycine Max L.) for the production of a composition intended to strengthen the mechanical strength of the hair and improve the appearance of the hair. The present invention relates to a peptide extract of soybean (Glycine Max L.) for increasing the expression of transglutaminases. The invention also relates to a cosmetic treatment method intended to strengthen the resistance of the hair and to combat external attacks on the hair, according to which a composition containing the active agent is applied topically to the areas to be treated.

Description

FIELD OF THE INVENTION PEPTIDE EXTRACT OF SOYBEANS AND ITS COSMETIC USE TO STRENGTHEN THE STRUCTURE OF THE HAVE Field of the Invention The present invention is in the field of cosmetics. The invention relates to the cosmetic use of a peptide extract of soy (Glycine Max L.), derived from the hydrolysis of soybeans, as an agent for strengthening the structure of the hair. The invention also relates to the cosmetic use of a soy peptide extract (Glycine Max L.) for the production of a composition intended to enhance the mechanical strength of the hair and improve the appearance of the hair. The present invention relates to a peptide extract of soybean (Glycine Max L.) for increasing the expression of transglutaminases. The invention also relates to a cosmetic treatment method intended to strengthen the resistance of the hair and to combat external attacks on the hair, according to which a composition containing the active agent is applied topically to the areas to be treated. BACKGROUND OF THE INVENTION The hair is keratinous appendages in the same way as the hairs, the eyelashes, the eyebrows or the nails. They have a physiological role of protection of the scalp, but especially a social role, and this, for a long time in the history of the man. Hair growth and renewal are mainly determined by the activity of hair follicles and their matrix environment. The hair follicle, also called "hair follicle" is a complex autonomous skin appendix that has six major compartments, some of dermal origin (conjunctive sheath and dermal papilla), others of epithelial nature (internal and external epithelial sheaths, hair shaft and sebaceous). At the base of the follicle is the matrix, at the origin of the three concentric layers of the hair. The hair consists of differentiated epithelial cells containing mainly keratins, themselves organized into very stable protein superstructures. The organization and strength of the hair shaft and hair are determined by the activity and good health of the follicle. It has been well described that the differentiation mechanisms of the keratinocytes of the epidermis and the hair follicle are substantially different. Thus, it is known that keratins of the hair shaft represent a family of keratins distinct from that expressed in the epidermis (Langbein et al., 2001, J. Biol Chem 276). Among these keratins, K6irs (Porter et al., 2001, Br.

J. Dermatol. 145: 558-568) is expressed in the inner sheath of the hair follicle but not in the epidermis, whereas the markers of epidermal differentiation, such as keratin K1 and K10, are not expressed in the hair follicle (Lenoir et al. 1988, Dev Biol 130: 610620).

During the different stages of differentiation, several families of proteins each have a specific function. Among them, transglutaminases participate in the crosslinking of proteins that will form the horny envelope of the cells of the hair shaft. Transglutaminases (EC 2.3.2.13) are a family of calcium-dependent enzymes that catalyze the formation of peptide bridges between a s-amino of a lysine residue and an γ-carboxamide of a glutamine residue. intramolecularly are extremely resistant to degradation (Lorand et al., Nat Rev Mol Cell Biol Feb, 4 (2), 2003). In humans, 9 transglutaminases (TG) have been identified, 4 of which are expressed in the skin and hair follicle. Transglutaminase-1 (TG1) is a catalytic enzyme linked to the keratinocyte membrane that contributes to the formation of the stratum corneum of the hair follicle. This layer acts as a mechanical barrier that protects the hair follicle and hair shaft from loss of water and infectious agents. This enzyme is directly involved in hair formation. Transglutaminase-3 (TG3) is expressed in hair follicles and in the late stages of keratinocyte differentiation. TG3, specific for the stratum corneum, is exclusively expressed in the hair shaft TG3 is a key enzyme in scaffolding and keratinisation of the stem. Transglutaminase-5 (TG5) is present in the upper layers of the epidermis and also plays a role in the early stages of epidermal differentiation. TG5 plays a key role in the physiological balance of skin and hair (Thibaut et al., Nature, 125: 3 September 2005).

TGs have very diverse substrates. Thus, they are able to crosslink the keratin with each other and with filaggrin, which leads to the stabilization and coordination of the keratin-filaggrin network in the horny cells. TGs allow the formation of bridges between protein precursors such as, for example, involucrine or trichohyaline, and thus promote keratinization.

The resistance and health of hair can be disrupted by many extrinsic and intrinsic factors. Among the intrinsic factors are the aging of the hair. During aging, there is less synthesis of proteins of the extracellular matrix (collagen, laminin, fibronectin), causing a loss of elasticity and tone of the subcutaneous tissue. There is also a decrease in the coherence and organization of the hair follicles (Courtois et al., 1995, Br. J. dermatol., 132: 86-93). The hair indeed loses its density, they are thinner and therefore more fragile. Regardless of the intrinsic aging, alterations of the hair or hair follicle can occur during external aggressions. In fact, if the hair is remarkably stable, certain external factors, such as the sun, responsible for photo-aging, free radicals, pollution or some inappropriate treatments can lead to premature deterioration of the hair structure. and exhaustion of the follicles. The term "external aggression" refers to 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 that are too detergent, stains and discolorations, permanent or too frequent straightening, and nail varnish removers. , nail polish, mechanical aggression, such as rubbing clothes, hairstyles causing repeated stretching, excessive brushing, brushing, drying in the air too hot. Pollution is understood to mean both pollution due to C.O.V. polluting particles in water, diesel particles, ozone or heavy metals, that pollution that may be due in particular to the solvent emissions of paints, glues, or wallpapers (such as toluene styrene, xylene or benzaldehyde), or even cigarette smoke. These external aggressions result in an alteration of the outer structure of the hair and its mechanical properties, but can also affect the hair follicle and cause premature aging. The cosmetic or pharmaceutical industry is always looking for compositions that make it possible to strengthen the strength and structure of the hair, exhibiting a rapid and lasting action over time.

The solution of the technical problem lies in the cosmetic use of a peptide extract of soy. The inventors have indeed demonstrated that a soy extract acts on the expression of transglutaminases, in the hair follicle and hair, thus promoting the strengthening of the structure of the hair shaft and its mechanical resistance to external aggressions.

Peptide extracts of soya have previously been described for their effects on the skin (FR 2904 552, FR 2887 772). The inventors have now demonstrated that such peptide hydrolyzates of soy have an activity on the strengthening of the structure of the hair follicle and an improvement of the health of the hair. It has been demonstrated in particular that the peptide hydrolyzate, when applied to scalp biopsies, increases the expression of the molecular markers of differentiation of keratinocytes of the inner sheath of the hair follicle, and by this action, can contribute to improve the structure of the follicle. The inventors have in fact demonstrated a cosmetic activity, a soy peptide extract derived from the hydrolysis of soy, capable of activating the transglutaminases expressed in the hair and / or the keratinous appendages. In particular, it has been demonstrated that these polypeptides or peptides, which constitute the peptide extract of soybeans, when applied to scalp biopsies in culture, significantly stimulate the expression of transglutaminases in the cells of the body. hair follicles. This has, among other things, been demonstrated by an increase in the expression of transglutaminases 1, 3 and 5. This new active ingredient capable of stimulating the expression of transglutaminases, and more generally capable of reinforcing the structure of the hair, thus allows 'open new cosmetic perspectives.

The invention and the advantages thereof will be better understood from reading the description. SUMMARY OF THE INVENTION The present invention firstly relates to the cosmetic use of a peptide extract of soy (Glycine Max L.), resulting from the hydrolysis of soybeans, as an agent for reinforcing the structure of the appendices. keratin. The human keratinous appendages to which the invention applies are in particular the hair, the eyebrows, the eyelashes, the beard hair, the mustache, the pubic hair and the nails. More specifically, the invention is applicable to human hair and / or eyelashes and / or nails.

In a particularly preferred embodiment, the invention relates to the cosmetic use of a peptide extract of soy (Glycine Max L.) to strengthen the structure of hair and hair follicles. In another preferred embodiment, the invention relates to the cosmetic use of a peptide extract of soy (Glycine Max L.) to strengthen the structure of the nails.

The soy extract used according to the invention offers the following advantages in particular: it increases the expression of transglutaminases; - it improves the structure and the guidance of the hair. - It increases the mechanical resistance of the hair to external aggressions.

The invention also relates to the cosmetic use of a soy extract for increasing the expression of transglutaminases in hair follicles and / or keratinous appendages. The characteristic molecular activity of the invention is defined in vitro by the ability of the active agent to increase the expression of transglutaminases, either by increasing the protein synthesis of transglutaminases (by direct or indirect modulation of the expression transglutaminase gene), or by other biological processes such as the stabilization of transglutaminase proteins or the stabilization of messenger RNA transcripts. Preferably, according to the present invention, the transglutaminases are transglutaminases 1, 3 or 5. The invention further relates to the cosmetic use of a soy extract to improve the structure and guidance of hair and / or hair follicles. The invention further relates to the cosmetic use of a soy extract for increasing the mechanical strength of hair and / or hair follicles.

The biological activity characteristic of the invention is defined in vitro by the ability of the active agent to reduce cell damage in hair follicles subjected to induced chemical stress. The term "capable of reinforcing the mechanical resistance of hair from external aggressions" means any substance of vegetable origin, and more particularly derived from soybean (Glycine Max L.), capable of exhibiting protective properties or reducing apoptosis. in hair follicle cells subjected to physicochemical or environmental stress. The term "activates transglutaminases" means any substance of vegetable origin, and more particularly derived from soy (Glycine Max L.), capable of increasing the expression of transglutaminases, or by the activation of protein synthesis ( by direct or indirect modulation of transglutaminase gene expression), either by increasing the biological activity of transglutaminases or by other biological processes such as the stabilization of transglutaminase proteins or the stabilization of transcripts of transglutaminases. Messenger RNA.

"Strengthening the structure and guidance of the hair" means any substance of vegetable origin, and more particularly from soy (Glycine Max L.), capable of increasing the expression or activity of the main active molecules present in the hair follicles and in the hair shaft, and more generally capable of increasing the principal functions of the hair follicles; namely, the keratinization and the organization of the layers of the hair rods. The term "peptide extract" is intended to mean a mixture of compounds predominantly represented by compounds of a peptide nature, in solution in a large volume of water or other polar solvents or a mixture of these solvents. The term "peptide-like compounds" means the protein fragments and the peptides present in the peptide extract according to the invention. The term "peptide" refers to a sequence of two or more amino acids linked together by peptide bonds or modified peptide bonds; the term "polypeptide" denotes a peptide of larger size. By the term "active agent" is meant "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.

By "topical application" is meant the application or spreading of the active agent according to the invention, or a composition containing it, on the surface of the skin or the keratinous appendages. By "physiologically acceptable" is meant any compound suitable for contact with the skin or mucosa without causing toxicity or intolerance reactions.

In the remainder of the presentation, the terms "active agent" and "soy extract" will be used interchangeably. The active agent according to the invention can be obtained by extraction of proteins of vegetable origin, followed by controlled hydrolysis which releases the biologically active peptide compounds.

The use of peptide extracts, and in particular peptide extracts of low molecular weight, has many advantages in cosmetics. In addition to generating peptidic compounds which did not previously exist in the starting protein mixture, hydrolysis and purification make it possible to obtain more stable mixtures, compositions that are more easily reproducible and that do not cause allergic reactions in cosmetics. .

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, and preferentially Glycine Max species is used. L.

According to the invention, the plant material used will be the grain and preferentially the grain removed from its envelope by a dehulling step. In a first step, the plant is ground 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). Any method of extraction or purification known to those skilled in the art can be used to prepare the extract according to the invention. For example, the controlled hydrolysis makes it possible to release compounds of peptide nature. It is possible, but not necessary to carry out the invention, either to extract the relevant proteins and then to hydrolyze them, or to carry out the hydrolysis on a crude extract and then to purify the compounds of peptidic nature afterwards. In a first step, the seeds contained in the seeds are crushed in order to obtain a powder or flour. The powder thus obtained can be previously treated with a cellulase to promote the elimination of sugars, and in particular insoluble polysaccharides. The proteins of the seed are then extracted according to the conventional method (modified Osborne, T. B., The Vegetable Proteins, 2nd Edition, Longmans, Green and Co., London, 1924); the plant mash is suspended in an alkaline solution containing an insoluble polyvinylpolypyrrolidone adsorbent material (PVPP) (0.01-20%); in fact, it is known that the hydrolyses and the subsequent purifications are facilitated by this means. In particular, the concentration of phenolic-type substances, interacting with proteins, is significantly reduced. The proteins can then be precipitated by varying the ionic strength or acidifying the medium, thereby eliminating soluble components and nucleic acids. The precipitate is then washed with an organic solvent such as, for example, ethanol or methanol, and the solvent is evaporated by drying in vacuo. The protein-rich precipitate is redissolved in water or other solvent and then constitutes a more purified form of the extract. 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 soluble fraction, containing proteins, carbohydrates and possibly lipids, is collected after centrifugation and filtration steps. This crude solution is then hydrolyzed under mild conditions to generate 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 among which we can then mention endoproteases of plant origin (papain, bromelain, ficin). According to a first embodiment of the invention, the hydrolyzed soy extract obtained in this step is used. For the same reasons as above, that is to say the elimination of polyphenol substances, an amount of polyvinylpolypyrrolidone can be added to the reaction medium during this mild hydrolysis step. The resulting extract can be further purified to select low molecular weight peptide compounds. The fractionation can advantageously be carried out by ultrafiltration and / or by a chromatographic type method. A dilution phase is then carried out in water or in any solvent mixture containing water. Thus, the active agent according to the invention is advantageously diluted in one or more physiologically acceptable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols. , cyclic polyols or any mixture of these solvents. The diluted active agent is then sterilized by ultrafiltration.

Thus, according to an advantageous form of the invention, the soy extract is diluted in one or more physiologically acceptable solvents, such as water, glycerol, ethanol, propanediol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. After this dilution, a peptide extract is obtained, characterized by a dry weight of 2 to 5 g / kg, a concentration of peptide-like compounds of 1 to 10 g / l, preferably of 1.5 to 3.5 g / l, a concentration of sugars of 0.05 to 1 g / l, preferably 0.1 to 0.3 g / l. Thus, according to an advantageous form of the invention, the soy extract has a dry weight of 2.5 g / kg and contains between 1.5 and 3.5 g / l of peptide-like compounds. The physicochemical characteristics and the content of protein and peptide compounds of the extract obtained according to the invention are analyzed qualitatively and quantitatively, according to standard techniques well known to those skilled in the art. The extract obtained is composed of peptides with a molecular weight of less than 5 kDa and is characterized by a concentration of sugars of less than 15%.

Thus, according to an advantageous form of the invention, the soy extract is a peptide extract in which the compounds of peptide nature have a molecular weight of less than 5 kDa. According to a second aspect, the present invention relates to a cosmetic care method comprising topical application to at least a portion of the scalp, eyelashes or nails, a peptide extract of soybean (Glycine Max .. ) resulting from the hydrolysis of soybean, as described above, in a composition comprising a physiologically acceptable medium, to obtain a reinforcing effect of the structure of the treated zones, and more particularly to improve the structure, the guidance and / or the mechanical resistance of the hair and / or the keratinous appendages. Advantageously, the soy extract is present at a concentration of between 0.0001% and 20% of the total weight of the composition, and preferably at a concentration of between 0.05% and 5% of the total weight of the composition, in a physiologically acceptable medium.

According to another advantageous embodiment of the invention, the active agent 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, carriers minerals such as talcs and bentonites.

The compositions for carrying out the invention may in particular be in the form of an aqueous, hydro-alcoholic or oily solution; an oil-in-water, water-in-oil emulsion or multiple emulsions; they can also be in the form of creams, suspensions or powders. 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 area to be treated in aerosol form. They can be used as a care product and / or as a makeup product for keratinous appendages, nails and / or hair. The compositions that may be used according to the invention may in particular consist of a shampoo, a conditioner, a pre-treatment or post-treatment treatment lotion that is aggressive for the hair, a cream or a styling gel, a restructuring lotion for the hair, a mask. etc. The composition may also be in the form of mascara for application to eyelashes, eyebrows or hair. The composition may also be in the form of nail polish or other composition capable of being applied to the nails.

These compositions may furthermore comprise any additive commonly used in the field of application envisaged 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. Advantageously, the composition that can be used to carry out the invention may comprise, in addition to the active agent according to the invention, at least one other active agent having cosmetic effects similar to and / or complementary to those of the invention. According to the invention, this active agent will be defined as an "additional active agent".

For example, the additional active agent (s) may be chosen from: anti-aging, lightening, moisturizing, draining, microcirculation promoting agents, pharmaceutical agents, exfoliants, desquamants, extracellular matrix stimulating, activating energy metabolism, antibacterials, antifungals , soothing, anti-radical, anti-UV, anti-acne, anti-inflammatory, anesthetic, providing a feeling of warmth, providing a feeling of freshness. Such additional agents may be chosen from the groups comprising: vitamin A and especially retinoic acid, retinol, retinol propionate, retinol palmitate, vitamin B3 and more particularly niacinamide, tocopherol niconitate, vitamin B5, vitamin B6, vitamin B12, panthenol, - vitamin C, in particular ascorbic acid, ascorbyl glucoside, ascorbyl tetrapalmitate, magnesium and sodium ascorbyl phosphate, - vitamins E, F, H , K, PP, coenzyme Q10, - inhibitors of metalloproteinase, or an activator of TIMPs, - DHEA, its precursors and derivatives, - amino acids such as arginine, ornithine, hydroxyproline, hydroxyproline dipalmitate, palmitoylglycine, hydroxylysine , methionine and its derivatives, amino acid compounds N-acyl, carnitine, carnosine, taurine, natural or synthetic peptides, including di-, tri-, tetra-, penta- and hexapeptides and their derivatives lipophylic are, isomers and complexed with other species such as a metal ion (e.g. copper, zinc, manganese, magnesium, and others). For example, the peptides commercially known under the name MATRIXYL®, ARGIRELINE®, COLLAXYL ™ VINCI 02TM PEPTIDE, CHRONOGENTM, LAMINIXYL ISTM, PEPTIDE Q10TM - plant peptide extracts such as extracts of soya, spelled, grapevine, rapeseed, flax , rice, corn, peas, dioscorea extract, guarana extract, ivy extract, fucus extract, algae extracts such as Palmata Palmaria, hydrolyzed Prunella extract vulgaris or elderberry extract, yeast extracts, extracts of Artemia Salina, dehydroacetic acid (DHA), compounds promoting hair growth and / or preventing hair loss, for example Minoxidyl, - oligosaccharides, polysaccharides, - phystosterols of synthetic or natural origin, - salicylic acid and its derivatives, alpha- and beta-hydroxy acids, silanols, - amino sugars, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-Dglucosamine, mannosami ne, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine, - extracts of polyphenols, isoflavones, flavonoids, such as grape extracts, pine extracts, olive extracts, - theine, theophylline, the theobromine, forskolin, esculin, lipids such as ceramides or phospholipids, oils of animal origin, such as squalene or squalane; vegetable oils, such as sweet almond oil, coconut oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower oil, wheat germ oil, corn germ oil, soya bean oil, of cotton, alfalfa, poppy, pumpkin, evening primrose, millet, barley, rye, safflower, passionflower, hazelnut, palm, apricot kernel, avocado, calendula ; ethoxylated vegetable oils, shea butter, - all UV screens and sunscreens, - cyclic AMP and its derivatives, - the activating agents of the adenylate cyclase enzyme, the phosphodiesterase enzyme inhibiting agents, the inhibitors of the enzyme ACE The composition usable according to the invention may be applied by any suitable route, including external topical, and the formulation of the compositions will be adapted by those skilled in the art. Advantageously, the compositions according to the invention are in a form suitable for topical application. These compositions must therefore contain a physiologically acceptable medium, that is to say compatible with the skin and the keratinous appendages, and cover all cosmetic forms.

It is obvious that the invention is directed to mammals in general, and more particularly to humans. 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. Figure 1: Transglutaminase activity in the keratinocytes derived from the hair follicle treated for 48 hours with 1% of the soy extract according to the invention.

Figure 2: Protection of hair follicles subjected to stress (treatment at 1% SDS for 24 hours) and treated or not with 1% of the soy extract according to the invention.

EXAMPLE 1 Preparation of a peptide hydrolyzate enriched in bioactive peptides from soybean meal (Glycine Max L.) The active principle 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. The cakes are the solid residues obtained after extraction of the oil from the soya beans. They represent 50 to 75% of the seed mass. 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% of 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); preferably 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 precipitated by varying the ionic strength in a neutral or acidic medium to remove soluble carbohydrate components, lipids, and nucleic acids. The medium is brought to 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 in vacuo. At this stage, about 50 grams of light yellow powder of crude protein extract are obtained containing: - Proteins: 75% - Carbohydrates: 20% - Lipids: 5% The precipitate rich in proteins 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% polyvinylpolypyrrolidone (POLYCLAR V ISP) and cysteine endopeptidases (papain, ficin). The purification of the extract thus obtained is continued by successive filtrations using Seitz-Orion plate filters of decreasing porosity (up to 0.2 μm) in order to obtain a brilliant and clear solution. After this series of filtration, 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 peptide-like compounds determined by the method of Lowry , between 0.1 and 5 g / l and preferably between 0.5 and 2 g / l. The protein profile of this extract is analyzed by electrophoresis gel. Under the same conditions as described above, two large families of proteins are observed: the first family, which is a minority, corresponds to proteins of molecular weight of 25 to 20 kDa, and the second family, which is very much in the majority, corresponds to proteins of lower molecular weight. at 5kDa. This extract is then purified by removing proteins of molecular weight greater than 5 kDa by tangential flow filtration steps. For this, the soy extract is pumped under pressure through a Pellicon® support equipped with Pellicon® 2 Biomax 30 kDa cassette. The first filtrate obtained is recovered to be filtered again through another Pellicon® 2 Biomax 5 kDa cassette. At the end of purification, a peptide extract of soybean is obtained, having a pH of between 4 and 7, and preferably between 5 and 6. It is characterized by a dry weight of between 1 to 8 g / l and preferably of 2 at 5 g / l, a content of compounds of peptidic nature is between 0.1 and 5 g / l, and preferably between 0.5 and 2 g / l, its sugar content between 0.5 and 2.5 g / l. A dilution phase is then carried out in a water-glycerol mixture in order to obtain a peptide extract characterized by a dry weight of 2 to 5 g / kg, and preferably of 2.5 g / kg, a concentration of peptide nature of 1.5 to 3.5 g / l, a sugar concentration of 0.1 to 0.3 g / l (ie less than 15%) and a polyphenol concentration of less than 1%. This purified and diluted extract corresponds to the peptide extract of soy according to the invention. It is characterized in that the compounds of peptide nature have a molecular weight of less than 5 kDa. EXAMPLE 2 Demonstration of the Increase in the Global Enzymatic Activity of Transglutaminases by the Hydrolyzate According to Example 1 The aim of this study is to determine the influence of the hydrolyzate according to Example 1 on the total transglutaminase activity in keratinocytes isolated from human hair follicles (KORS). For this purpose, the total enzymatic activity of the transglutaminases is determined spectrophotometrically using a fluorescein-labeled amino acid donor substrate.

Protocol: KORS are seeded in 96-well black plates. After 4 days of culture, the KORS are treated with a 1% solution of hydrolyzate according to Example 1 for 48 hours (the active ingredient is added every 24 hours). A positive control is achieved by treatment of cells with EGCG at 1.1g / ml (epigallocatechin gallate, the main polyphenol of green tea). The substrate used is fluorescein-labeled cadaverine (Invitrogen A10466) diluted to 100 μl in culture medium. The substrate is incubated for 2 hours with the cells. The cells are then rinsed twice in HBSS buffer and fixed with a mixture of acetic acid-ethanol-water (1:49:50) for 20 minutes. After two rinses in ethanol and then three rinses in HBSS buffer, a spectrofluorometer reading at excitation wavelengths of 485 nm and emission of 530 nm is carried out. Under these conditions, the transglutaminase activity is proportional to the amount of fluorescence emitted (expressed in fluorescence units), relative to the total amount of proteins present in each well, previously assayed by the BCA technique. Results: The results are shown in histogram form in Figure 1. The results are expressed as a percentage of the untreated control. In the presence of 1% of hydrolyzate according to Example 1, the enzymatic activity is increased by 55.6% after 48 hours. Conclusion: The hydrolyzate according to Example 1 significantly increases the total enzymatic activity of transglutaminases in keratinocytes isolated from human hair follicles. EXAMPLE 3 Demonstration of the Activating Effect of the Hydrolyzate According to Example 1 on the Expression of TG1, TG3 and TG5 The aim of this study is to determine the influence of the hydrolyzate according to Example 1 on the expression of the different transglutaminases expressed in the human hair follicle. For this purpose, specific immunofluorescent markings were performed on cultured keratinocytes isolated from hair follicles (KORS) and on microdissected hair follicles of the human scalp. Protocol for immunostaining on keratinocytes isolated from hair follicle (KORS) in culture: KORS in culture were treated with a 1% hydrolyzate solution according to Example 1 for 24 hours. For anti-TG1 immunostaining, the cells are washed and fixed with 3.7% paraformaldehyde for 10 minutes. The cells are then incubated in the presence of an anti-TG1 specific antibody (Santa Cruz sc-25786, polyclonal rabbit), and then a suitable secondary antibody, coupled to a fluorescent marker. For the other immunolabelings, the cells are washed and fixed with cold methanol for 4 minutes. The cells are then incubated in the presence of a specific antibody; anti TG3 (Abcam ab53236, monoclonal mouse) or anti TG5 (Abcam ab26992, polyclonal rabbit). The slides are then observed under an epifluorescence microscope (Nikon Eclipse E 80i microscope), after mounting in an ad hoc environment. Immunolabeling Protocol for Microdissected Hair Follicles: Microdissected hair follicles of the human scalp are cultured in a suitable medium. A 1% solution of hydrolyzate according to Example 1 is applied topically for 24 hours. For labeling of TG1, the microdissected hair follicles are then embedded in resin and frozen in liquid nitrogen. Sections of about 6 μm are then made by cryostat. Immunostaining is performed using a specific antibody; anti TG1 (Clinisciences BT-621, monoclonal mouse) then a suitable secondary antibody, coupled to a fluorescent marker. The skin sections are then examined under an Epi-fluorescence microscope (Nikon Eclipse E 80i microscope). For TG3 markings, the microdissected hair follicles are embedded in paraffin and histological sections 3 μm in thickness are made. The slides are deparaffinized, hydrated and then subjected to immunolabeling with an antibody directed against TG3 (Abcam ab53236, monoclonal mouse) and then a suitable secondary antibody, coupled to a fluorescent marker. The skin sections are then examined under an Epifluorescence microscope (Nikon Eclipse E 80i microscope). Results: In all the conditions tested, more intense fluorescence was observed in the cultures and sections of hair follicles treated with the hydrolyzate according to Example 1 at 1% than under the control conditions, untreated. Conclusions: The hydrolyzate according to Example 1 stimulates the expression of TG1, TG3 and TG5 in keratinocytes isolated from hair follicle in culture. The hydrolyzate according to Example 1 stimulates the expression of TG1 and TG3 in microdissected hair follicles cultured ex vivo.

EXAMPLE 4 Demonstration of the Activating Effect of the Hydrolyzate According to Example 1 on the Expression of the Structural Proteins of the Internal Layer of the Hair Follicle The aim of this study is to determine the influence of the hydrolyzate according to Example 1 on the structural proteins of the inner sheath of the hair follicle. For this, the expression of the main markers of differentiation, specifically expressed in the keratinocytes of the inner sheath of the hair follicle, was studied. The markers tested are keratin 71, desmocollin-1, trichohyaline, and involucrine. On the other hand, desmocollin-1 is a constituent of desmosomal cell junctions, whereas involucrine and trichohyaline are substrates of transglutaminases.

Protocol for immunolabeling on microdissected hair follicles: Repeat the protocol of Example 3 for the immunolabeling of TGasel: immunostaining with an antibody directed against keratin 71 (PROGEN, GP-K6irsl, Guinea pig polyclonal) or trichohyaline ( Santa Cruz, sc-80607, monoclonal mouse), or desmocollin-1 (Santa Cruz, sc-18117, polyclonal goat), or the involucrine (Novocastra NCL-INV, SY5 clone, monoclonal mouse) or TG1 (BT Clinisciences -621, monoclonal mouse). A suitable secondary antibody coupled to a fluorescent label is then used. For ease of observation, the nuclei of the cells can be counter-stained by DAPI (4 ', 6'-di-Amidino-2-phenylindole, a blue fluorescent molecule capable of binding strongly to DNA). After mounting in an appropriate medium, the slides are observed under an epifluorescence microscope (Nikon Eclipse E 80i microscope). Results: In all the conditions tested by immunofluorescence, more intense fluorescence was observed on the sections of hair follicles treated with the hydrolyzate according to Example 1 at 1% than under the control conditions, untreated. The image analysis makes it possible to evaluate the fluorescence intensity and consequently the increase in the expression of the markers tested. The increase of trichohyaline is of the order of 55%, involucrin of 32% and desmocollin-1 is of the order of 116%, keratin-71: 45% Conclusions: The hydrolyzate according to Example 1 stimulates the expression of keratin 71, involucrine, trichohyaline, in human keratinocytes of the inner sheath of the human hair follicle. The hydrolyzate according to Example 1 also improves the cohesion of the inner sheath of the hair follicle and strengthens the internal structure of the hair follicle which could be favorable to improving the mechanical strength of the hair.

EXAMPLE 5 Demonstration of the protective effect vis-à-vis the external aggressions of the hydrolyzate according to Example 1 The purpose of this study is to determine the protective effect on the skin of the scalp and hair follicle of the hydrolyzate according to Example 1 with respect to external aggressions. For this, an ex vivo model of severe aggression of the cutaneous barrier was used. Protocol: Biopsies of human scalp skin containing hair follicles are pre-treated with the 1% hydrolyzate for 24 hours, while others receive only placebo (PBS buffer). These biopsies are then subjected to an attack caused by a sodium lauryl sulphate (SDS) detergent, applied at 1% topically for 24 hours, while some of the biopsies are not subjected to stress. After incubation, the biopsies are embedded in the resin and frozen in liquid nitrogen. The histological staining with hematoxylin-eosin is then performed on frozen sections 61.1 m thick. Microscopic observation analysis is performed on the epidermis and follicular structures. Results: The histological sections of skin and hair follicles treated with the hydrolyzate according to Example 1 at a concentration of 1%, show a decrease in structural alterations caused by detergent stress (control), such as vacuoles or loss. cohesion between the different compartments of the hair follicle. The results concerning the hair follicles are shown in FIG. 2. Conclusion: The hydrolyzate according to Example 1 improves the internal structure of the hair root and preserves the structure of the scalp and the hair follicle from damage due to chemical stress. , by a detergent.

Example 6: Preparation of compositions 1 - Nourishing care for hair and scalp Apply the product on the moist scalp. Massage to evenly distribute the product. Strengthens hair while making it smooth and easy to comb.30 Formulation 1 2 INCI Name%% Mass Commercial Mass Supplier Phase A Deionized Water - Q.S. Q.S. Aminomethyl Propanol AMP-95 0.05 0.05 Acrylic Acid / VP Crosspolymer UltraThix ™ P-0.85 0.85 Ashland 100 Phase B Gycerol Dilaurate Emulsynt ™ 0.50 0.50 Ashland GDL Jojoba Seed Oil - 2.00 2.00 Lipo Cetearyl Alcohol - 2.00 2.00 Rita Phase Cyclopentasiloxane SiTecTM 0.50 1.00 Ashland CM040 Phase D Ashland VP / DMAPA Acrylates Copolymer StylezeTM CC- 3.00 3.00 10 Water 20.00 20.00 Aminomethyl Propanol 0 , 37 0.37 E phase GermabenTM 0.75 0.75 Ashland M Diazolidinyl Urea (and) Methylparaben (and) Propylene Glycol Hydrolysate according to Example 1 0.50 1.00 Ashland Total 100.00 100.00 Add to cart water and AMP-95 in a stirred vessel. Add UltraThixTM P-100 to vigorously stirred water and keep stirring for 30 minutes. Heat phase A to 65 ° C. Independently, heat the ingredients of phase B together at 65 ° C and mix them. Transfer phase B to the container of phase A and mix for 15 minutes. Start cooling the preparation to 35 ° C. At 45 °, add phase C to the main mixture. In a separate container, mix the ingredients from phase D together and add to the main mix while mixing. Add one by one the ingredients of phase E under 35 °.

Shake until a uniform appearance is obtained.15 2 - Serum to strengthen the structure of the hair: Apply the product on the wet scalp. Massage to evenly distribute the product. Promotes the growth or regrowth of hair and makes the hair more nervous. Formulation 1 2 INCI Name Name%% Mass Commercial Mass Supplier Water Q.S. Q.S. Hydroxyethylcellulose Natrosol 0.35 0.50 Ashland 250HEIR Disodium EDTA Dissolvine 0.05 0.05 Akzo Nobel NA-2S VP / DMAPA Acrylates Copolymer StylezeTM CC-5.00 5.00 Ashland 10 Quaternium-26 CeraphylTM 65 1.00 1, 00 Ashland Panthenol Ritapan DL 0.15 0.15 RITA Propylene Glycol (and) Diazolidinyl Urea (and) Iodopropynyl Butylcarbamate Liquid 0.50 0.50 Ashland GermallTM Plus Hydrolyzate according to Example 1 1.00 1.00 Ashland Total 100, 00 100.00 Disperse Natrosol 250HHR and Disodium EDTA in water with stirring. Heat to 50-60 ° C, and shake until uniform. Add StylezeTM CC-10, and shake until smooth. Allow to cool to room temperature and add the ingredients in the order listed by shaking until a uniform appearance is obtained between each addition. 3 - Non aerosol treating foam: Apply the product on the wet scalp. Massage to evenly distribute the product. Strengthens the vitality and health of the hair and promotes a lasting hold of the hairstyle, especially in a humid atmosphere. Formulation 1 2 INCI Name Name%% Mass Commercial Mass Supplier Water Q.S. Q.S. PEG-45M POLYOX N-0.075 0.075 Dow 750 Polyquaternium-55 (20%) StylezeTM W 5.00 5.00 Ashland Propylene Glycol (and) Diazolidinyl Urea (and) Iodopropynyl Butylcarbamate Liquid 0.50 0.50 Ashland GermallTM Plus PEG / PPG-25/25 Dimethicone WackerTM 0.30 - Ashland DMC 6031 Palmitamidopropyltrimonium Chloride Varisoft PATC - 0.80 Degussa Hydrolyzate according to Example 1 1.00 1.00 Ashland Total 100.00 100.00 Put water in a suitable container, and shake vigorously to create a vortex. Disperse the Polyox in the vortex and shake until completely dissolved. Add Styleze W-20 and shake until smooth. Then add the ingredients in the order listed by shaking until a uniform appearance is achieved between each addition.

Claims (12)

REVENDICATIONS1. Cosmetic use of a peptide extract of soy (Glycine Max L ..) resulting from the hydrolysis of soybeans, as an agent for reinforcing the structure of keratinous appendages. 2. Cosmetic use of a peptide extract of soy (Glycine Max L ..) according to claim 1, wherein the keratinous appendages are hair and hair follicles. 3. Cosmetic use of a peptide extract of soy (Glycine Max L ..) according to claim 1, wherein the keratinous appendages are the nails. 4. Cosmetic use according to claims 1 to 3, wherein the soy extract causes an increase in the expression of transglutaminases in the hair follicles and / or keratinous appendages. 5. Cosmetic use according to one of claims 1 to 4, wherein the soy extract improves the structure and guidance of hair and / or hair follicles. 6. Cosmetic use according to one of claims 1 to 5, wherein the soy extract increases the mechanical strength of hair and / or hair follicles. 7. Cosmetic use according to one of claims 1 to 6, wherein the soy extract is diluted in one or more physiologically acceptable solvents, selected from water, glycerol, ethanol, propanediol, butylene glycol , dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols or any mixture of these solvents. 8. Cosmetic use according to one of claims 1 to 7, wherein the soy extract contains between 1.5 and 3.5 g / 1 of peptide compounds. 9. Cosmetic use according to one of claims 1 to 8, wherein the soy extract is a peptide extract in which the peptide compounds have a molecular weight of less than 5 kDa. 10. Cosmetic treatment method comprising the topical application to at least a portion of the scalp, eyelashes or nails, a peptide extract of soy (Glycine Max L ..) resulting from the hydrolysis of soybean, such as as described in one of claims 1 to 9, in a composition comprising a physiologically acceptable medium, to obtain a reinforcing effect of the structure of the treated areas, and more particularly to improve the shape, the guidance and / or the mechanical strength hair and / or integuments. 11. Cosmetic care method according to claim 10 wherein the soy extract is present at a concentration of between 0.0001% and 20% of the total weight of the composition, and preferably at a concentration of between 0.05% and 5% of the total weight of the composition. 12. Cosmetic care method according to one of claims 10 or 11 wherein the composition further comprises at least one additional active agent selected from vitamin A, retinoic acid, retinol, vitamin B3, vitamin B5, vitamin B6, vitamin B12, vitamin C, vitamin E, vitamin F, vitamin H, vitamin K, vitamin PP, coenzyme Q10, metalloproteinase inhibitors, amino acids, carnitine, carnosine, taurine, natural or synthetic peptides, plant peptide extracts, yeast extracts, extracts of Artemia Salina, minoxidil, synthetic or natural phystosterols, salicylic acid, oligosaccharides, polysaccharides, amino sugars, polyphenols, flavonoids, lipids, phospholipids, cyclic AMP and its derivatives, the activating agents of the adenylate cyclase enzyme, the phosphodiesterase enzyme inhibiting agents, the thein e, theophylline, theobromine, forskolin, esculin, ACE inhibitors, dioscorea extract, guarana extract, ivy extract, fucus extract, seaweed extracts such as Palmata Palmaria, hydrolysed extract of Prunella vulgaris or elderberry extract.