Novel use of a peptide to improve the extracellular matrix of the skin's dermis and / or mucous membranes

Abstract

Novel Use of a Peptide to Improve the Extracellular Matrix of the Dermis of Skin and / or Mucous Membranes. The present invention relates to the non-therapeutic cosmetic use of the peptide with sequence SEQ ID No. 1 and / or its homologs and / or salts and / or derivatives, particularly in the form of rice hydrolysate enriched with this peptide or fermentate enriched with this peptide, to improve the ECM fibers of the dermis of skin and / or mucous membranes. The present invention also relates to the applications of this peptide in cosmetic and / or dermatological compositions.

Description

Title of the invention: Novel use of a peptide to improve the extracellular matrix of the dermis of the skin and / or mucous membranes technical field

[0001] The present invention relates to the use of a peptide with SEQ ID No. 1 sequence and / or its homologs, derivatives, and / or salts, optionally in a fermentate of a microorganism producing it or in a protein hydrolysate, to improve the extracellular matrix fibers of the dermis of the skin and / or mucous membranes, in particular the flexibility and / or density of the skin, including the scalp, and / or mucous membranes, and / or the anchoring of skin appendages, preferably hair, notably to prevent and / or reduce hair loss. The present invention also relates to the applications of this peptide in cosmetic and / or dermatological compositions. Previous technique

[0002] The extracellular matrix (ECM) plays an essential role in the structure of tissues in the human and animal body, particularly through its functions of support, adhesion, and regulation of cellular exchange. The ECM is largely composed of glycoproteins, proteins, and glycosaminoglycans. These molecules are synthesized in their native form within cells in contact with the ECM. From the intracellular compartment, they are secreted outside the cells. After maturation processes, they organize and arrange themselves into a network of fibers that forms the ECM. In this fibrillar form, they are then in their functional form.

[0003] Exposure of the skin to aggressive environmental agents such as temperature variations, particularly heat and cold, humidity, dry air, sunlight, and / or pollution, activates proteases responsible for the degradation of the dermal extracellular matrix (ECM), notably by reducing the number of collagen and elastic fibers in the dermis. This results in a loss of the skin's general properties, particularly those of the dermis, such as density and elasticity. ECM molecules are therefore the subject of intense study in the cosmetic and pharmaceutical fields, where numerous ingredients aim to prevent and compensate for this degradation in order to improve the overall condition of the dermis.

[0004] One of the classic approaches consists of inhibiting proteases, thereby preventing the degradation of elastic fibers. However, this purely prophylactic action does not restore the properties of the ECM once these have been reduced. A second approach then consists of stimulating protein synthesis Pro-collagen I and elastin are synthesized by fibroblasts. However, simply increasing these syntheses in cultured cells in vitro does not necessarily translate into a greater quantity of functional molecules, i.e., molecules organized within the extracellular matrix (ECM). This approach is therefore insufficient to restore certain skin properties, particularly flexibility and density.

[0005] Restoring these properties requires obtaining a specific organization and arrangement of the dermal ECM fibers, which involves numerous partners. Indeed, with regard to collagen, the formation of triple helix structures for collagen (tropocollagen) and its secretion outside the cell into the extracellular matrix, as well as the assembly and organization of collagen triple helices into fibrils, which themselves organize into larger mature fibers, are necessary to maintain and restore a functional ECM in the dermis and the properties of the dermis.

[0006] Similarly, the formation of mature elastic fibers capable of stretching is necessary to maintain a functional ECM in the dermis. Elastin is synthesized as soluble tropoelastin by fibroblasts and acquires its physicochemical properties, including stretchability and return to its initial state after stretching, following its cross-linking by enzymes of the lysyl oxidase family and its deposition onto fibrillin-rich microfibrils in the extracellular matrix to form mature elastic fibers. In addition to fibrillins, and in particular fibrillin-1, fibulin-5, EMILIN-1, and MFAP4 (Microfibril Associated Protein 4) are also particularly important in the formation of these mature elastic fibers.

[0007] The present invention aims to provide a new cosmetic and / or dermatological active ingredient to improve the ECM fibers of the dermis, particularly of the scalp and mucous membranes. The present invention also aims to provide a new cosmetic and / or dermatological active ingredient to improve the density and / or flexibility of the skin and / or mucous membranes. The present invention further aims to provide a new cosmetic and / or dermatological active ingredient for the specific care and / or treatment needs of skin and / or mucous membranes with weakened and / or altered dermal ECM, particularly under the effect of aggressive agents, which is notably the case for sensitive and / or sensitized skin and / or mucous membranes.

[0008] Surprisingly and unexpectedly, the inventors discovered that the SEQ ID No. 1 peptide, its homologs, salts and / or derivatives are capable of improving the ECM fibers of the dermis.

[0009] The peptide from SEQ ID No. 1 has already been described in the field of cosmetics among a large number of peptides derived from rice and peas in patent application WO2017009484A1 (as SEQ ID No. 349) as an anti-inflammatory capable of inhibiting the secretion of TNFalpha by macrophages in the presence of lipopolysaccharides, as well as in patent application WO2017009490Al (as SEQ ID No. 245) as an agent capable of increasing cell proliferation in the basal lamina of the epidermis and having a mitotic effect on the basal lamina of the epidermis and thus treating the visible signs of aging.

[0010] This peptide is naturally present in a protein of the rice grain Oryza saliva, from which it can be obtained. However, it is not naturally present in sufficient quantity to be detected and produce the properties of which the invention is a subject. In the context of the present invention, it has been obtained by chemical synthesis and by a biotechnological production process using the yeast Saccharomyces cerevisiae. However, it can also be obtained by enzymatic hydrolysis of rice grain as described in patent applications WO2017009484A1 and WO2017009490A1.

[0011] While a large number of rice peptide hydrolysates are available on the cosmetics market, they differ greatly in their profile and composition, particularly in the different protein fragments they contain, which significantly impacts their biological properties. In protein hydrolysis, the bonds linking the various peptides and amino acids are cleaved using various conventional methods in this field, such as acids, bases, or catalysis. However, these different techniques lead to protein hydrolysates with highly variable compositions, sometimes even resulting in free amino acids. Chemical hydrolysis using acids or bases, being non-specific, typically leads to lower-quality products, particularly in terms of color, odor, and / or stability.Enzymatic hydrolysis, due to its specificity, offers the advantage of being able to be carried out under controlled conditions, particularly pH, temperature, and pressure. However, here again, the enzymes are chosen to obtain the peptides of interest, leading to the production of specific hydrolysates that differ significantly in terms of the peptide fragments they contain, whether in terms of fragment nature, molecular weight, or distribution.

[0012] These rice hydrolysates of all kinds are used in the field of cosmetics for applications for skin and / or hair care.

[0013] However, due to their nonspecific or directed methods of obtaining other peptides, all these hydrolysates differ greatly in their compositions and properties and none describes or contains the peptide according to the invention in sufficient quantity to allow obtaining the properties according to the invention.

[0014] The invention thus relates to the non-therapeutic cosmetic use of the SEQ ID No. 1 peptide, its homologs, derivatives and / or salts to improve the ECM fibers of the dermis of healthy skin and / or healthy mucous membranes.

[0015] This peptide has many additional advantages in that it also increases collagen 1 synthesis by fibroblasts, and reduces inflammation by inhibiting the release of TNF alpha by macrophages and by inhibiting the release of interleukin-8 by keratinocytes.

[0016] The peptide according to the invention, by its complementary properties, provides a perfectly adapted and complete solution for the care and / or treatment of sensitive and / or sensitized skin and / or mucous membranes.

[0017] Moreover, all these complementary properties also make it an excellent cosmetic care and / or treatment agent for the skin and / or mucous membranes and / or healthy skin appendages of the body and face.

[0018] The SEQ ID No. 1 peptide according to the invention can be obtained by chemical synthesis using conventional methods. However, according to a preferred embodiment of the invention, the SEQ ID No. 1 peptide is obtained by synthesis using a recombinant microorganism, preferably a bacterium such as Escherichia coli or a yeast of the genus Pichia or Saccharomyces, preferably S. cerevisiae. Particularly advantageously, the SEQ ID No. 1 peptide is used in association with the fermentate of the microorganism producing it.

[0019] Alternatively, the peptide according to the invention is in the form of a hydrolysate of Oryza saliva rice proteins, advantageously with a peptide content of at least equal to or greater than 0.00001% by dry weight relative to the total weight of the hydrolysate, preferably between 0.0001% and 10%, more preferably between 0.001% and 3%, by dry weight relative to the total weight of the hydrolysate.

[0020] The peptide according to the invention is the SEQ ID No. 1 peptide: TVFDGVLRPGQL

[0021] This peptide contains the 12 amino acids: Tyr Val Phe Asp Gly Val Leu Arg Pro Gly Gin Leu and has an average molecular weight of 1301.51 g / mol.

[0022] This 12-amino-acid sequence is naturally present in rice grain proteins (Oryza saliva), and the peptide can be obtained by enzymatic hydrolysis of rice grains as described in patent applications WO2017009484A1 and WO2017009490A1, or by chemical or biotechnological synthesis using conventional methods. It can also be purified or obtained as shown in Example 1.

[0023] According to the invention, "peptide" means any isolated amino acid sequence, natural or synthetic, which may be obtained by chemical or biotechnological synthesis or by extraction from a biological tissue, for example a plant, an animal or a microorganism, in particular yeast, expressing naturally or after transduction the amino acid sequence, possibly after post-translational modification thereof.

[0024] The term "derivative" means the peptide modified by the addition of a functional group, for example coupled with a hydrophilic or hydrophobic penetrating agent, a stabilizing agent, or a protecting group, according to conventional methods known to those skilled in the art, in particular by acylation of the terminal -NH2 group, or amidation or alkylation of the terminal -COOH group. This may, in particular, involve grafting onto the free -NH2 group of the last amino acid a carboxyl group comprising 1 to 24 carbons, saturated or unsaturated, linear or branched, and which may also contain -OH, -SH, COOH, or CONH2 groups.It may also involve grafting an amide group (-CONH2) or an alkoxy group onto the free -COOH group of the last amino acid, said group comprising 1 to 24 carbons, saturated or unsaturated, linear or branched and possibly also containing -OH, -SH, COOH or CONH2 groups.

[0025] The chemical synthesis techniques of peptides are known to those skilled in the art and examples include the techniques described in the references JM Stewart and JD Young, solid phase peptide synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984), as well as M. Bodanzsky and Bodanzsky, The practice of peptide synthesis, Springer Verlag, New York (1984).

[0026] According to the invention, the term "homologous" of an amino acid sequence means an amino acid sequence having at least 85%, preferably 90%, in particular at least 95%, and more particularly at least 98%, preferably at least 99%, sequence identity with said sequence and possessing biological activity of the same nature. Sequence homology can be identified by any conventional technique in the field, such as, for example, via the BLAST computer interface available on the NCBI website at http: / / blast.ncbi.nlm.nih.gov configured with the default parameters.

[0027] A homolog of an amino acid sequence may differ from that sequence, for example, by one or more deletions and / or insertions, and / or one or more substitutions of an amino acid. According to one embodiment, a homolog of an amino acid sequence may comprise one or more conservative amino acid substitutions. A conservative substitution is the replacement, in a sequence, of an amino acid by another amino acid having substantially similar physicochemical properties, or properties sufficiently close to those of the original amino acid, so that the properties and functions of the peptide are not, or are not substantially, affected. The modifications to the amino acid sequences described above may be generally referred to as "mutations." Thus, the homologs of the peptides according to the invention also relate to the mutants and variants of the amino acid sequences of the invention having biological activity of the same nature. By "biological activity of the same nature" of an amino acid sequence of the invention, we mean in particular its ability to exhibit the properties according to the invention.

[0028] According to the invention, "salts" refers to the peptide comprising a counterion from the acid used during cleavage after its chemical synthesis. The salts of the peptide according to the invention include trifluoroacetate, acetate, and hydrogen chloride.

[0029] The peptide according to the invention can thus be in the form of its homolog, its salt, and / or its derivative. Preferably, the peptide according to the invention is exactly that of SEQ 1.

[0030] The amino acids constituting the peptide according to the invention can be in their L, D or DL ​​forms. Preferably, they are all in the L configuration.

[0031] According to the invention, "improving the fibers of the dermal extracellular matrix (ECM)" means improving the structure and / or organization and / or density of the collagen and / or elastic fibers of the dermal extracellular matrix. This improvement is measured by an increase in the number of functional fibers in the dermal extracellular matrix and results in an increase in the flexibility and / or density properties of the skin and / or mucous membranes. Thus, advantageously, the use according to the invention is to increase the density and / or flexibility of healthy skin and / or mucous membranes. These properties can be measured by conventional methods known to those skilled in the art. The following techniques may be cited as examples: - Flexibility: This property can be measured using conventional techniques, notably by measuring the elastic modulus (or Young's modulus), for example on a reconstructed skin model. The improvement of the dermal ECM fibers then results in a decrease in the elastic modulus value measured after treatment with the peptide according to the invention, compared to the value obtained without treatment. Such a method is described in Example 4. - Density: This property can be measured using conventional techniques, notably by determining the total dermal density by measuring the echogenicity of ultrasound in the dermis. The improvement in the density of the dermal ECM fiber network results in an increase in the total dermal density value measured after skin treatment with the peptide according to the invention, compared to untreated skin. Such a method is described in Example 6.

[0032] According to the invention, the improvement of the extracellular matrix fibers of the dermis does not include the increase in collagen and elastin synthesis, which is not sufficient on its own to induce an improvement in the extracellular matrix fibers, particularly for sensitive and / or sensitized skin. Furthermore, preferably, the use according to the present invention is not for the prevention and / or treatment of signs of aging.

[0033] According to the invention, the improvement of the extracellular matrix fibers of the dermis is measured by the increase in the amount of fibrillin-1 in the ECM fibers of the dermis and / or by the increase in the amount of EMILIN-1 in the ECM fibers of the dermis and / or by the increase in the amount of collagen fibers in the extracellular matrix of the dermis. Thus, advantageously, the use according to the invention is to increase the amount of fibrillin-1 in the ECM fibers of healthy dermis and / or to increase the amount of EMILIN-1 in the ECM fibers of healthy dermis and / or to increase the amount of collagen fibers in the extracellular matrix of healthy dermis.The use according to the invention is also further to increase the amount of fibulin-5 synthesized by fibroblasts in the healthy dermis and / or to increase the amount of MFAP4 synthesized by fibroblasts in the healthy dermis and therefore to increase the synthesis of fibulin-5 and / or MFAP4 by fibroblasts in the healthy dermis.

[0034] The amount of fibrillin 1 in the dermal ECM can be measured by immunolabeling in a reconstructed skin model, as described for example in Example 2, and the increase can be evaluated in the presence of the peptide according to the invention by comparison with the measurement carried out in the absence of this peptide.

[0035] The amount of EMILIN-1 in the ECM of the dermis can be measured by immunolabeling in a reconstructed skin model, as described for example in Example 2, and the increase can be evaluated in the presence of the peptide according to the invention by comparison with the measurement carried out in the absence of the peptide.

[0036] The quantity of ECM collagen fibers can be measured by measuring the intensity of a second harmonic signal by confocal photon microscopy, as described in Example 3.

[0037] The amounts of fibulin-5 and / or MFAP4 synthesized by dermal fibroblasts can be measured by immunolabeling on a lysate of dermal fibroblasts after culture, as described for example in Example 5, and the increase can be evaluated after culture in the presence of the peptide according to the invention by comparison with the measurement carried out after culture in the absence of the peptide.

[0038] The peptide according to the invention is also suitable for improving the anchoring of hair follicles, in particular hair. The peptide according to the invention thus makes it possible to prevent and / or reduce hair loss, particularly of hair.

[0039] For the purposes of the present invention, "phaneres" means nails and "keratin fibers".

[0040] The term “keratin fibers” means hair fibers (hair), eyelashes, eyebrows, hairs, particularly those of the nasal mucosa, ears, beard and / or moustache.

[0041] The peptide according to the invention is therefore particularly suitable for the care and / or treatment of sensitive or sensitized skin and / or mucous membranes.

[0042] Thus advantageously the use according to the invention is intended for the care of sensitive and / or sensitized skin and / or mucous membranes.

[0043] Generally speaking, "sensitive skin and / or mucous membranes" can be defined as healthy skin and / or mucous membranes that, by their nature, have a very low tolerance for aggressive agents, particularly environmental agents such as pollutants, climatic factors (wind, cold, heat), UV exposure, emotional factors such as stress, and / or chemical agents (heavy metals, detergents, compounds contained in cosmetic treatments such as perfumes, preservatives, alcohols, pH adjusters, AHAs, or dermatological agents such as retinoic acid (vitamin A acid)), and / or aggressive conditions such as perspiration and mechanical aggressions such as hair removal, shaving, friction, and even water, especially hard water. Sensitive skin is not pathological, unlike allergic skin.However, they can react to aggressive agents and / or conditions with unsightly and / or uncomfortable cutaneous and / or mucosal manifestations such as dryness of the skin and / or mucous membranes, loss of evenness in skin tone and / or mucous membranes, particularly through the appearance of redness, sensations of tightness, tingling, prickling, prickling, tension and / or itching, a rough texture of the skin and / or mucous membranes and / or a loss of softness to the touch. Thus, the "sensitive skin" characteristic can be assessed by the individual themselves through subjective skin sensations or by the dermatologist through objective skin reactions.

[0044] Most of the time, sensitive skin and / or mucous membranes will manifest themselves in well-defined locations such as, for example, the scalp, face, skin folds, buttocks in infants, etc. It can therefore be a matter of sensitive skin and / or mucous membrane areas.

[0045] Similarly, "sensitized skin and / or mucous membranes" are healthy skin and / or mucous membranes made momentarily sensitive and therefore not pathological as such.

[0046] For the purposes of this invention, "cosmetic" means a non-pharmaceutical, non-therapeutic use that is not intended for the prevention and / or treatment of skin and / or mucous membranes and / or hair and nails that have been diagnosed as pathological by a specialist in the field, such as a dermatologist. It therefore refers to use on healthy skin and / or mucous membranes and / or hair and nails.

[0047] The term “healthy skin, in particular healthy scalp and / or healthy mucous membranes and / or healthy hair and nails” means all or part of an area of ​​skin, in particular the scalp and / or mucous membranes and / or healthy hair and nails, in particular human skin, on which the peptide according to the invention is applied and which is said to be “non-pathological” by a dermatologist, i.e., not showing any cancer, infection, scar, disease or skin condition such as candidiasis, impetigo, psoriasis, eczema, acne, ichthyosis, gingivitis or dermatitis or wounds or injuries or canker sores or ulceration or burns and / or other dermatoses, or aphthous ulcers or inflammation (in particular sunburn) or irritation,or urticaria or allergy such as contact allergy or pathologies associated with a decrease in ECM fibers and / or a decrease in Fibrillin-1 content and / or EMILIN-1 content and / or a decrease in the amount of collagen fibers and / or any combination thereof, in particular solar elastosis. Advantageously, healthy skin and / or mucous membranes according to the invention are not susceptible to developing a pathology associated with a decrease in ECM fibers and / or a decrease in Fibrillin-1 content and / or EMILIN-1 content and / or a decrease in the amount of collagen fibers and / or any combination thereof, in particular solar elastosis.

[0048] For the purposes of this invention, "skin" means the skin of all or part of the body, particularly human skin, including the scalp, selected from the face, hands, arms, décolletage, legs, neck, back, shoulders, abdomen, wrists, forearms, ankles, thighs, nape of the neck, scalp, joint and / or armpit creases, labial mucous membranes, preferably the face, particularly the area around the eyes and / or mouth and the forehead. In particular, skin is that which may be exposed to aggressions, especially those likely to degrade the fibers of the extracellular matrix (ECM), particularly areas exposed to aggressive environmental or chemical agents, friction, and / or maceration. This includes, more specifically, the face, scalp, hands, neck, décolletage, joint creases, and / or armpits.

[0049] For the purposes of the present invention, "mucous membrane(s)" means the ocular, nasal, vaginal, anal and / or oral mucosa, in particular the oral labial mucosa, preferably the labial, ocular and / or nasal mucosa.

[0050] Preferably, the application is by topical route, advantageously on specific parts and / or areas of the body chosen from among the scalp, face, hands, arms, décolleté, legs, neck, back, shoulders, belly, wrists, forearms, ankles, thighs, nape of the neck, joint folds, and / or armpits, labial mucous membranes.

[0051] The peptide according to the invention is topically acceptable. For the purposes of the present invention, "topically acceptable" means an ingredient suitable for a topical application, non-toxic, non-irritating to the skin and / or mucous membranes and / or hair, which does not induce an allergic response and is not chemically unstable.

[0052] The peptide according to the invention can be obtained by chemical synthesis, by biotechnological synthesis using a naturally produced or recombinant microorganism and / or by enzymatic hydrolysis of biological material containing in particular a microorganism, of plant or animal nature.

[0053] Thus, according to one embodiment of the present invention, the peptide is used in the form of a rice protein hydrolysate (Oryza saliva), that is to say a peptide hydrolysate of Oryza saliva, in particular enriched in peptide according to the invention, preferably in liquid form.

[0054] According to an alternative embodiment of the present invention, the peptide is used in the form of a microorganism fermentate by producing, preferably from a recombinant yeast, more preferably chosen from recombinant yeasts of the genus Pichia or Saccharomyces, even more preferably from the yeast Saccharomyces cerevisiae, in particular genetically modified to produce this sequence, said fermentate being preferably enriched in peptide according to the invention.

[0055] According to an alternative embodiment of the present invention, the peptide is used in a purified form from a hydrolysate or a fermentate by the usual techniques of peptide purification, in particular selected from membrane filtration, chromatography, precipitation and / or immunoprecipitation.

[0056] According to the invention, "hydrolysate" means the product of chemical and / or enzymatic hydrolysis of biological material containing and / or producing, naturally or by genetic modification, the peptide according to the invention, such as animal, vegetable or microorganism material, in particular yeast.

[0057] According to the invention, "fermentate" means the fermented must produced by a microorganism that produces, naturally or through genetic modification, the peptide according to the invention, in particular yeast or bacteria. The fermentate thus corresponds to the culture medium containing the nutrients necessary for the growth and multiplication of the microorganism, as well as the fermentation products excreted by the microorganism or released during the lysis of the microorganism.

[0058] According to the invention, the term “the peptide-enriched hydrolysate or fermentate according to the invention” means a hydrolysate or fermentate in which the peptide of SEQ ID No. 1, according to the invention, is detected at a level exceeding the limit of detection with sensitive techniques in this area, for example LC-MS, that is to say generally a level at least equal to or greater than 0.00001% by weight of peptide relative to the total dry weight of the hydrolysate or fermentate.

[0059] According to an alternative embodiment of the invention, the SEQ ID No. 1 peptide is contained in a hydrolysate, preferably in an amount of at least equal to or greater than 0.00001% by weight of dry matter relative to the total weight of the hydrolysate, preferably between 0.0001% and 10%, more preferably between 0.001% and 1%, by weight of dry matter relative to the total weight of the hydrolysate.

[0060] According to a preferred embodiment, the rice peptide hydrolysate (or protein hydrolysate) is obtained by enzymatic hydrolysis in a single step. According to an alternative embodiment, the hydrolysate can be obtained by successive hydrolysis using the same enzyme or a mixture of enzymes.

[0061] Enzymatic hydrolysis is carried out by one or more proteolytic enzymes, which may be proteases of plant origin or from microorganisms.

[0062] According to one embodiment, the enzymatic hydrolysis takes place until its completion, which can be determined by a person skilled in the art in a known manner, for example by determining the constant pH or by photometry using the detection of free NH2 groups or by determining the constant amount of SEQ ID No. 1 peptides by liquid chromatography and mass spectrometry.

[0063] The amount of enzyme(s) used is not critical in itself, but should be in the range of 0.05 to 5%, preferably 0.1 to 2% by weight relative to the weight of the starting material containing the peptide.

[0064] The hydrolysates obtained can then be further processed, for example by filtering the undissolved fractions. For better stabilization, the pH is preferably set at values ​​between 3.0 and 7.5, preferably between 3.5 and 5.5.

[0065] Alternatively, the peptide according to the invention can be obtained by chemical synthesis using conventional techniques in this field, in particular those described. Advantageously, the peptide according to the invention will have a purity level of at least 50%, preferably at least 70%, advantageously at least 80%, preferably at least 90%, and even more preferably at least 95% (w / w).

[0066] According to another alternative method, the peptide according to the invention can be obtained by biotechnological synthesis, in particular from a recombinant bacterial or yeast strain, i.e. genetically modified to produce the peptide according to the invention.

[0067] The yeast strain is preferentially chosen from Pichia and Saccharomyces, in particular Saccharomyces cerevisiae.

[0068] The production of the peptide by a yeast strain classically comprises the following steps: - transformation of yeast using a peptide expression system according to the invention coupled to export signal sequences, typically of plasmid type; - culture of the transgenic yeast on a culture medium; - batch and / or fed-batch fermentation; - recovery of fermentation must and removal of yeast biomass, for example by centrifugation; - Concentration and / or separation of the peptide.

[0069] Preferably, the peptide according to the invention is produced as described in example 1).

[0070] In a particularly advantageous embodiment, the peptide is contained in the fermentate and used directly in this form.

[0071] In one embodiment, the hydrolysates or fermentates obtained are in liquid form, in particular aqueous solutions, and can be used directly or in concentrated form; they preferably have a dry matter content in the range of 1 to 50% by weight, preferably from 5 to 30% by weight relative to the total weight of the hydrolysate or fermentate. Additives such as polyols, glycols, acids (citric acid, sorbic acid, sulfuric acid, benzoic acid or their salts...) may be added.

[0072] The hydrolysates or fermentates obtained at this stage can be further concentrated and / or purified to select the targeted molecular weight fractions, by successive steps of ultrafiltration or nanofiltration through filters of variable porosity, or by means of a chromatographic-type method, for example to specifically enrich the fermentate or hydrolysate in these peptides.

[0073] It is also possible to produce the peptide hydrolysates or fermentates according to the invention in powder form by drying, (atomizer, lyophilized,....) with or without a support such as mannitol, maltodextrin, cyclodextrin, according to the classic formulation techniques known to those skilled in the art.

[0074] According to one embodiment, the preferred hydrolysate or fermentate according to the invention contains the SEQ ID No. 1 peptide, in a total content of at least equal to or greater than 0.00001% by weight relative to the total dry weight of the hydrolysate or fermentate, preferably between 0.0001% and 10%, more preferably between 0.001% and 3%, even more preferably between 0.01% and 1%.

[0075] The average molecular weight expressed in daltons (or g / mol) is determined by size exclusion chromatography known to those skilled in the art.

[0076] The rice peptide hydrolysate according to the invention can be obtained by carrying out the following steps: - solubilization of rice protein, or of a rice protein isolate containing at least 70%, preferably at least 80% protein, in water at a temperature above 70°C for pasteurization; - protein hydrolysis; preferably the hydrolysis is carried out enzymatically using proteolytic enzymes, preferably of plant origin or derived from microorganisms, by selecting the enzyme or enzymes and adjusting the temperature and pH conditions to obtain the correct degree of hydrolysis and molecular weight profile; - inactivation of the enzyme(s), preferably by heat treatment: this inactivation is carried out according to the technical recommendation of the supplier(s) of the enzyme(s); - separation of soluble and insoluble phases, by centrifugation and / or filtration, and recovery of the soluble phase containing the peptides; - possibly a concentration step to increase the dry matter content, followed by a filtration step to recover the soluble phase containing the peptides; - obtaining a filtrate, the peptide-enriched rice protein hydrolysate according to the invention, which is in liquid form and which also constitutes an embodiment of the invention.

[0077] The steps of the processes described above, taken individually, are common in the field of protein hydrolysates and a person skilled in the art is able to adjust the reaction parameters according to their general knowledge to obtain the rice protein hydrolysate according to the invention.

[0078] The peptide according to the invention, pure or in the form of peptide-enriched hydrolysate or peptide-enriched fermentate, is used topically.

[0079] According to one embodiment, it can be used: - in the form of a cosmetic or pharmaceutical ingredient intended to be incorporated into a cosmetic or pharmaceutical composition, and further comprising a suitable cosmetic or pharmaceutical vehicle, or - in the form of a cosmetic or pharmaceutical composition comprising it and further advantageously comprising a suitable cosmetic or pharmaceutical vehicle.

[0080] For the purposes of the present invention, "topical route" means the application of the peptide according to the invention, preferably in the form of a hydrolysate or fermentate enriched in peptide and / or of the composition and / or of the ingredient according to the invention on the surface of the skin, in particular the scalp and / or mucous membranes, and / or hair, preferably hair, in particular by direct application or by spraying.

[0081] For the purposes of the present invention, "cosmetic and / or pharmaceutical ingredient(s)" means one or more plant extracts and / or one or more natural or synthetic molecules and / or mixtures thereof intended for cosmetic application and / or pharmaceutical. Cosmetic ingredients are notably defined by the International Nomenclature of Cosmetic Ingredients (INCI).

[0082] For the purposes of the present invention, the term “suitable cosmetic or pharmaceutical vehicle” means that its composition or components are suitable for use in contact with human skin and / or mucous membranes without undue toxicity, incompatibility, instability, allergic response, or their equivalents.

[0083] The cosmetic or pharmaceutical ingredient, in particular dermatological in liquid form containing the peptide according to the invention, in particular in the form of a peptide-enriched fermentate according to the invention and in particular that of Example IC, can be used in a cosmetic or pharmaceutical composition, in particular dermatological, preferably at a dry matter weight content relative to the total weight of the composition of between 0.01 and 10%, advantageously between 0.1 and 5%, in particular between 0.2 and 3%.

[0084] In one embodiment of the invention, the peptide according to the invention (the SEQ ID No. 1 peptide and / or its homologs, derivatives and / or salts) is in the form of a cosmetic composition comprising it and advantageously further comprising a suitable cosmetic vehicle.

[0085] In another embodiment of the invention, the peptide according to the invention, in particular the SEQ ID No. 1 peptide, preferably in the form of the peptide-enriched fermentate, is present in the cosmetic or pharmaceutical composition at a level of between 1x10⁶% and 10% by weight, preferably from 1x10⁵% to 0.1% by weight, or even more advantageously from 1x10⁴% to 0.01% by weight of dry matter relative to the total weight of the composition.

[0086] The compositions according to the invention may contain any suitable solvent and / or any suitable vehicle and / or any suitable excipient, possibly in combination with other compounds of interest.They may, in particular, contain a cosmetically or dermatologically acceptable excipient and / or a suitable cosmetic or pharmaceutical vehicle selected from surfactants, preservatives, buffering agents, swelling agents, chelating agents, biocidal agents, denaturants, opacifying agents, pH adjusters, reducing agents, stabilizing agents, emulsifiers, thickeners, gelling agents, film-forming polymers, solvents, fillers, bactericides, odor absorbers, mattifying agents, conditioning agents, texturizing agents, gloss-enhancing agents, pigments, colorants, perfumes and chemical or mineral sunscreens, trace elements, essential oils, sweeteners, and flavor modifiers. These combinations are also covered by the present invention. The CTFA Cosmetic Ingredient Handbook, Second Edition (1992) describes... various cosmetic and pharmaceutical ingredients commonly used in the cosmetic and pharmaceutical industry, which are particularly suited for oral administration and / or topical use.

[0087] Advantageously, the excipient(s) and / or vehicle(s) are selected from the group comprising polyglycerols, esters, polymers and cellulose derivatives, lanolin derivatives, phospholipids, lactoferrins, lactoperoxidases, sucrose-based stabilizers, vitamin E and its derivatives, xanthan gums, natural and synthetic waxes, vegetable oils, triglycerides, unsaponifiables, phytosterols, silicones, protein hydrolysates, betaines, aminoxides, plant extracts, sucrose esters, titanium dioxide, glycines, and parabens, and preferably from the group consisting of steareth-2, steareth-21, glycol-15 stearyl ether, cetearyl alcohol, phenoxyethanol, methylparaben, ethylparaben, propylparaben, butylparaben, butylene glycol, caprylyl glycol, natural tocopherols, glycerin, dihydroxycetyl sodium phosphate,isopropyl hydroxycetyl ether, glycol stearate, triisononanoine, octyl cocoate, polyacrylamide, isoparaffin, laureth-7, a carbomer, propylene glycol, hexylene glycol, glycerol, bisabolol, a dimethicone, sodium hydroxide, PEG 30-dipolyhydroxystearate, caprylic / capric triglycerides, cetearyl octanoate, dibutyl adipate, grapeseed oil, jojoba oil, magnesium sulfate, EDTA, a cyclomethicone, xanthan gum, citric acid, sodium lauryl sulfate, waxes and mineral oils, isostearyl isostearate, propylene glycol dipelargonate, propylene glycol isostearate, PEG 8, beeswax, hydrogenated palm kernel oil glycerides, lanolin oil, sesame oil, cetyl lactate, lanolin alcohol, castor oil, titanium dioxide, lactose, sucrose, low-density polyethylene, isotonic saline solution, and mixtures thereof.

[0088] The cosmetic or pharmaceutical composition or peptide, preferably the hydrolysate or peptide-enriched fermentate according to the invention, possibly in the form of a cosmetic or pharmaceutical ingredient, may be in any of the dosage forms conventionally used for topical application, such as liquid or solid forms, or even in the form of a pressurized liquid. In particular, they may be formulated as a solution, aqueous or oily, an aqueous or oily cream or gel, especially in a jar or tube, including a shower gel, shampoo, lotion, emulsion, hydrogel, microemulsion or nanoemulsion, including oil-in-water, water-in-oil, multiple, or silicone-based emulsion, serum, lotion, especially in a glass or plastic bottle or dosing bottle or aerosol, ampoule, liquid soap, paste, dermatological bar, ointment, foam, aerosol, mask, etc. A patch is an anhydrous product, preferably liquid, paste, or solid, for example, in stick form or as a powder, particularly makeup. Specifically, the composition is presented as a serum, lotion, cream, milk, ointment, paste, foam, emulsion, hydrogel, shower gel, mask, patch stick, or makeup powder, advantageously as a cream or lotion. Preferably, the composition is presented as a cream or lotion for the skin and / or hair, or as a shampoo or conditioner, rinse-off or leave-on.

[0089] The peptide according to the present invention, preferably the peptide-enriched fermentate according to the invention, has the advantage of not irritating and / or stripping the skin and / or mucous membranes, allowing the maintenance of the sebum level and is particularly suitable for the care and / or treatment of sensitive and / or sensitized skin and / or mucous membranes.

[0090] In the case of administration via the mucosal route, the cosmetic or pharmaceutical composition or the peptide according to the invention, possibly in the form of a cosmetic or pharmaceutical ingredient, may be in the form of an eye drop, a lotion, an aerosol, a gel, or a mucoadhesive composition.

[0091] The cosmetic or pharmaceutical composition may also include other cosmetic or pharmaceutical ingredients, active on the treatment of the skin and / or mucous membranes and / or hair, inducing a complementary or synergistic effect with the peptide according to the invention, chosen for example from the so-called anti-aging active cosmetic ingredients.

[0092] Among the active anti-aging cosmetic ingredients, we can mention: - those to prevent the appearance of pigmentation and / or increase radiance of the complexion, including an extract of the mushroom Inonotus obliquas marketed under the name Inolixir™ by the applicant, an extract of Argania spinosa oil marketed under the name Arganyl™ by the applicant, an extract of Moringa oleifera seeds marketed under the name Purisoft™ by the applicant, a combination of an extract of Salvia miltiorrhiza and niacinamide marketed under the name CollRepair™ by the applicant, an extract of Achillea millefolium marketed under the name Neurobiox™ by the applicant, an extract of Cassia alata leaves marketed under the name DN-Age™ and / or an extract of lychee marketed under the name Litchiderm™ as antioxidant actives, and an extract of chicory marketed under the name Lox-Age™, a yeast extract marketed under the name Vitacell™,an extract of Polygonum bistorta marketed under the name Perlaura™, an extract of galangal marketed under the name Hyalufix™, an extract of corn marketed under the name Deliner™ or , an extract of Voandzeia subterranea marketed under the name Epigenist™ by the plaintiff; - those promoting skin firmness, in particular by action on collagen, such as a synthetic tetrapeptide marketed under the name Dermican™, an extract of Hibiscus abelmoschus marketed under the name Linefactor™, a purified pea extract marketed under the name Proteasyl™, an extract of Manilkara multinervis marketed under the name Elestan™, an extract of Khaya senegalensis marketed under the name Collalift™18, an extract of Argan pulp marketed under the name Argassential™, an extract of Schizandra chinensis marketed under the name Sqisandryl™ by the Applicant, retinol, vitamin C, an extract of Davilla rugosa marketed under the name Collguard™, an extract of hydrolyzed soy protein marketed under the name Phytokine™.

[0093] The peptide according to the invention can also be used in association with cosmetic ingredients active on sensitive skin, in particular the protein extract of ungerminated deoiled Moringa oleifera seed marketed under the trade name Purisoft®, a plant extract of Cestrum latifolium marketed under the name Symbiocell ™, a butter extracted from the fruit of the Irvingia gabonensis tree marketed under the name Irwinol™, an extract of Eperua falcata root marketed under the name Eperuline™, an N-acetyl-L-Tyrosyl-L-Prolyl-L-Phenylalaninamide peptide (INCI: Acetyl Tetrapeptide 15) marketed under the name Skinasensyl™, an extract of Inonotus obliquus marketed under the name Inolixir™ by the Applicant.

[0094] The peptide according to the present invention may also be used in combination with ingredients active on the cutaneous and / or mucosal microbial flora and / or active on the skin barrier function, in particular moisturizing and / or soothing agents, including an oligosaccharide obtained by enzymatic synthesis marketed by Solabia under the name BioEcolia™ or an alpha-glucohydrate complex marketed by the same company under the name Ecoskin™, an extract of Alisma plantago-aquatica, an extract of Argania spinosa (Lipofructyl™ Argan), a mixture of ceramides (Sphingoceryl™ VEG), purifying extracts of Boldo (Betapur™), products based on inulin or fructooligosaccharides, extracts of bifidobacteria, or an extract of Orthosiphon stamineus to combat oily skin (MAT-XS™ Bright), a natural extract of honey marketed by the Plaintiff under the name Melhydran™ for its hydrating properties,a flax extract marketed under the name Oligolin™ by the Plaintiff, a yeast extract modified by biotechnology and marketed by the Plaintiff under the name Relipidium™, a Pueraria lobata root extract marketed under, the name Inhipase™ by the Applicant, a beta-glucan derivative from baker's yeast marketed by Mibelle under the name CM-Glucan Forte™ and / or an extract of Mirabilis jalapa marketed under the name Pacifeel™ by Sederma.

[0095] The present invention also relates to a method of non-therapeutic cosmetic care and / or treatment comprising the topical application, in particular daily, to at least one area of ​​healthy skin, advantageously of healthy scalp, and / or of healthy mucous membranes and / or of healthy hair, of the peptide according to the invention and / or of its homologs, salts and / or derivatives, preferably in the form of a peptide-enriched fermentate according to the invention or of a cosmetic composition according to the invention, to improve the ECM fibers of the dermis of healthy skin and / or healthy mucous membranes, in particular the structure and / or organization and / or density of the collagen and / or elastic fibers of the extracellular matrix of the dermis, more particularly to increase the density and / or flexibility of healthy skin and / or healthy mucous membranes.

[0096] Advantageously, the skin and / or mucous membrane area is chosen from among the scalp, face, hands, arms, décolletage, legs, neck, back, shoulders, abdomen, wrists, forearms, ankles, thighs, nape of the neck, joint folds and / or armpits, labial mucous membranes.

[0097] Advantageously, the invention also relates to a cosmetic treatment method for improving the ECM fibers of the dermis of healthy skin, and / or healthy mucous membranes of an individual who needs / desires it, comprising the steps: - The identification on the individual of an area of ​​healthy skin, and / or healthy mucous membranes, whose dermal ECM fibers one wishes to improve, and - The topical application on this area of ​​healthy skin and / or healthy mucous membranes, of the peptide according to the invention or of a cosmetic composition containing the peptide according to the invention in an amount effective to improve the ECM fibers of the dermis, in particular in a content of SEQ ID No. 1 peptide and / or its homologues, salts and / or derivatives of between 1x10 6% to 10% by weight, preferably from 1x10 5% to 0.1% by weight, more advantageously from 1x10 4% to 0.01% by weight, of dry matter in relation to the total weight of the composition.

[0098] The present invention also relates to the SEQ ID No. 1 peptide and / or its homologs and / or its salts and / or its derivatives according to the present invention, preferably in the form of a peptide-enriched fermentate according to the invention, for its use alone or in a pharmaceutical composition, particularly a dermatological one, comprising it, in the treatment and / or prevention of at least one pathology associated with a decrease in ECM fibers and / or a decrease of the Fibrillin-1 content and / or the EMILIN-1 content and / or a decrease in the amount of collagen fibers and / or any combination thereof, such as solar elastosis.

[0099] Advantageously, the peptide is present in a pharmaceutical composition, in particular dermatological, in a content of between 1x10 6% to 10% by weight, preferably from 1x10 5% to 0.1% by weight, more advantageously from 1x10 4% to 0.01% by weight, of dry matter in relation to the total weight of the composition, said composition further advantageously comprising an appropriate pharmaceutical vehicle, advantageously an appropriate dermatological vehicle.

[0100] Other objects, features and advantages of the invention will become clear to those skilled in the art upon reading the explanatory description which refers to examples and figures which are given only as examples and which shall in no way limit the scope of the invention.

[0101] The examples form an integral part of the present invention and any feature which appears new in relation to any prior art from the description taken as a whole, including the examples, forms an integral part of the invention in its function and in its generality.

[0102] Thus, each example has a general scope.

[0103] On the other hand, in the examples, and unless otherwise indicated, the temperature is expressed in degrees Celsius and the pressure is atmospheric pressure. Brief description of the drawings

[0104] [Fig.1A-1B-1C] Figures 1A-1B-1C represent 3 photographs obtained by confocal microscopy of a histological section of reconstructed skin after immunostaining with filaggrin (in green): 1A-untreated control; 1B-after treatment with 0.00001% (0.1 pg / ml) of peptide according to the invention; 1C-after treatment with 0.0001% (1 pg / ml) of peptide according to the invention.

[0105] [Fig. 2A-2B-2C] Figures 2A-2B-2C represent three photographs obtained by confocal microscopy of a histological section of reconstructed skin after immunostaining of EMILIN-1 (in green): 2A-untreated control; 2B-after treatment with 0.00001% (0.1 pg / ml) of peptide according to the invention; 2C- after treatment with 0.0001% (1 pg / ml) of peptide according to the invention.

[0106] [Fig. 3A-3B] Figures 3A-3B represent two photographs obtained by Multiphoton confocal microscopy of a reconstructed dermis and visualization of the second harmonic signal (SHG) (in white): 3A-untreated control; 3B- after treatment with 0.0001% (1 pg / ml) of peptide according to the invention.

[0107] Example 1: Different methods of obtaining the SEP ID No. 1 peptide TVFDGVLRPGOL

[0108] LA ) From a plant hydrolysate:

[0109] This 12-amino-acid peptide can be obtained from a rice protein hydrolysate (Oryza .saliva) obtained by enzymatic hydrolysis with a single proteinase or as described in patent application WO2017009484A1 for obtaining SEQ ID No. 349 or as described in patent application WO2017009490Al for obtaining SEQ ID No. 245.

[0110] LB) By chemical synthesis:

[0111] The peptide according to the invention was obtained by chemical synthesis at a purity of 99.4% determined by HPLS-UV. The identity of the peptide according to the invention was confirmed by LC-MS by the spectrum of the peptide containing 1305.5 (M+H+) and 651 (M+2H+) ions.

[0112] LC) From a recombinant microorganism fermentate

[0113] A peptide expression system according to the invention, coupled to export signal sequences, was constructed in the yeast Saccharomyces cerevisiae for the production and secretion of the peptide into the culture medium. The transgenic yeast was cultured on a standard medium (SY-2) containing glucose as a carbon source, ammonium sulfate as a nitrogen source, and minerals (KH2PO4, MgSO4, Na2SO4, NaCl, CaCl2) as well as vitamins, and having a pH of 4.1. After pre-culturing for 12 to 16 hours on the standard SY-2 medium to achieve an optical density (OD) of 5-8, a bioreactor was inoculated with a 7% (w / w) inoculation rate in the standard SY-2 medium relative to the total weight of yeast and culture medium. The culture was conducted at 30°C, the pH was regulated to 4, the partial pressure of oxygen (pO2) was regulated to a minimum of 30% by action on agitation and a maximum aeration of 0.3 WM.After a 1-hour batch fermentation phase, a 37-hour fed-batch fermentation phase was conducted with the addition of glucose and ammonium sulfate. After 48 hours of culture, the fermentation broth was collected and centrifuged to recover the supernatant and remove yeast biomass. This supernatant was concentrated 5- to 10-fold by evaporation under vacuum at 25°C, and activated carbon was added at a concentration of 25% (w / w) of activated carbon relative to the dry mass of the concentrate. After pre-filtration to remove the activated carbon and filtration through a 0.22 µm filter, the resulting liquid was lyophilized to obtain a powder with a peptide content according to the invention of 2984 pg / g, determined by HPLC-UV. The identity of the peptide was confirmed by LC-MS by the spectrum of the peptide containing the ions 1305.5 (M+H+) and 651 (M+2H+).

[0114] Example 2: Evaluation of the properties of the peptide according to the invention on fibrillin-1 and on EMILIN-1

[0115] A 3D reconstructed skin model (Mimeskin) was used to evaluate the performance of the peptide according to the invention obtained by chemical synthesis as described in Example 1B). A quantity of 2 x 10⁶ fibroblasts was seeded onto a dermal culture medium containing a mixture of collagen, glycosaminoglycans (GAGs), and chitosan (Mimedisc) and cultured by immersion at 37°C with 5% CO₂ for 56 days for the dermal building phase. Then, 10⁶ keratinocytes were seeded and cultured for 7 days for the keratinocyte proliferation phase. Subsequently, the models were placed at the air-liquid interface for the keratinocyte differentiation phase, and the culture was continued for 14 days. The cultures were treated or not (untreated control) with the peptide according to the invention in aqueous solution at a dose of 0.1 qg / mL or 1 qg / mL from day 2 and every 2 days except when the cells were seeded.Each condition was performed with n = 4 (4 replicates). Histological sections were prepared for each condition and each replicate using a cryostat. After fixation, the sections were immunostained for the detection of fibrillin-1 and EMILIN-1 by fluorescence (Figs. 1A-1B-1C and 2A-2B-2C, in green). Evans blue was used as a counterstain for visualizing the overall morphology (in red). Observations were performed using a confocal laser scanning microscope (TCS-SP2, Leica), and the percentage of surface area occupied by the targeted protein on the dermal surface was quantified by image analysis. The results are expressed as the mean percentage compared to the untreated control standardized to 100%. Statistical analysis was performed versus the untreated control after verifying the normality of the distribution of values ​​(Shapiro-Wilk test) with Student's t-test. The significance level was set at 5%.

[0116] After treatment with doses of 0.1 qg / ml and 1 qg / ml, the peptide according to the invention stimulated fibrillin-1 by +69% and +87%, respectively. After treatment with doses of 0.1 qg / ml and 1 qg / ml, the peptide according to the invention stimulated EMILIN-1 by +19% and +27%, respectively.

[0117] This increase in the fibrillin-1 microfibril network in the extracellular matrix is ​​clearly observable in Figures 1A-1B-1C.

[0118] Figures 2A-2B-2C illustrate the increase in EMILIN-1, which is a microfibril ligand involved in their correct organization and functionality in the extracellular matrix.

[0119] The peptide according to the invention therefore makes it possible to increase the content of fibrillin 1 and Emilin-1 and thus the organization of elastic microfibrils.

[0120] Example 3 Evaluation of the properties of the peptide according to the invention on collagen fibers.

[0121] A 3D reconstructed dermis model (Mimederm) was used to evaluate the performance of the peptide obtained by chemical synthesis. 2 x 10⁶ fibroblasts were seeded onto a dermal culture medium containing a mixture of collagen, glycosaminoglycans (GAGs), and chitosan (Mimedisc) and cultured by immersion at 37°C with 5% CO₂ for 56 days. The cultures were treated or untreated (untreated control) with the peptide according to the invention obtained by chemical synthesis (described in Example 1B) in aqueous solution at a dose of 1 µg / mL starting on day 2 and every 2 days thereafter. Each condition was performed in n = 3 (3 replicates). A multiphoton confocal microscopy analysis enabling the generation of a second harmonic signal (SHG) was performed on each condition and each replicate.

[0122] SHG imaging is a specific technique for observing collagen fibrils and fibers in the extracellular matrix. Collagen organized into fibrils has a non-centrosymmetric structure that is necessary to produce an SHG signal. The SHG signal is visualized in white in Figures 3A-3B (autofluorescence of surface cells and the reconstructed skin scaffold visualized in green).

[0123] Results are expressed as a mean percentage compared to the untreated control standardized to 100%. Statistical analysis was performed versus the untreated control after verifying the normality of the distribution of values ​​(Shapiro-Wilk test) using Student's t-test. The significance level was set at 5%.

[0124] Figures 3A-3B illustrate the increased presence of organized collagen fibers in the extracellular matrix of the dermis. A significantly higher intensity of the SHG signal is observed in Figure 3B, highlighting a greater density of collagen fibers in the extracellular matrix when the peptide according to the invention was present. Analysis of the volume occupied by the SHG signal also shows a 68% increase in the quantity of these fibers in the extracellular matrix in the presence of the peptide according to the invention compared to the untreated control.

[0125] Example 4: Evaluation of the properties of the peptide according to the invention on the flexibility of the dermis

[0126] A 3D reconstructed skin model (Mimeskin) was used to evaluate the performance of the peptide according to the invention obtained by fermentation of recombinant yeast as described in Example LC. 2 x 10⁶ fibroblasts were seeded onto a dermal culture medium containing a mixture of collagen, glycosaminoglycans (GAGs), and chitosan (Mimedisc) and cultured by immersion at 37°C with 5% CO₂ for 57 days for the dermis-building phase. Then, 10⁶ keratinocytes were seeded and cultured for 7 days for the keratinocyte proliferation phase. Next, the models were placed at the air-liquid interface for the keratinocyte differentiation phase, and the culture was continued. for 14 days. Cultures were treated or untreated (untreated control) with the peptide according to the invention in aqueous solution at a dose of 0.8 qg / mL or 3 qg / mL starting on day 2 and every 2 days thereafter, except when cells were seeded. Each condition was performed in n = 4 (4 replicates). For each condition and each replicate, 3 histological sections were prepared using a cryostat, and elastic modulus measurements were performed by atomic force microscopy (AFM) on the dermis with a 0.5 N / m conical probe. The results are expressed as the average value across the different cryosections. The elastic modulus (or Young's modulus) is the quantity that measures a material's resistance to elastic deformation when a force is applied to it. The higher the elastic modulus of the material, the stiffer it is. Conversely, the lower it is, the more flexible the material is.A Student's t-test was performed and showed results significantly different from the result measured with the untreated model (p<0.001) for both doses tested. The Young's modulus values ​​in the absence (untreated) or in the presence of the peptide according to the invention are summarized in Table 1 below.

[0127] [Tables] Average Conditions (in Pascals) Untreated 15.1 kPa Peptide 0.8 qg / ml 7.7 kPa Peptide at 3 qg / ml 3.4 kPa

[0128] The results show an improvement in the flexibility of the dermis.

[0129] Example 5: Evaluation of the properties of the peptide according to the invention on fibulin-5 and on MFAP4

[0130] Mature and functional elastic fibers in the ECM require the assembly of various glycoproteins, including fibulin-5 and MFAP4 (Microfibril Associated Protein 4). A human dermal fibroblast culture assay was used to evaluate the performance of the peptide according to the invention, obtained by synthesis as described in Example 1B), on the synthesis of these glycoproteins.

[0131] Human dermal fibroblasts were seeded at 5 x 10⁴ cells / cm² and cultured with a defined medium (FGM) by immersion at 37°C with 5% CO₂ until confluence. Once confluence was reached, the cells were incubated for 48 hours in the presence or absence (untreated control) of the peptide according to the invention (example 1B) in aqueous solution at doses of 0.1 qg / mL and 1 qg / mL. The cells were then recovered and lysed to measure the fibulin-5 content in the lysate. The culture medium was recovered to measure the MFAP4 content. The concentration in the samples was determined by a BCA assay to adjust the loading of each sample to the same protein concentration for Western blot analysis. Fibulin-5 and MFAP4 were analyzed by capillary electrophoresis after incubation with a primary antibody, immunostaining with a secondary antibody, and chemiluminescence measurement. Each condition was performed in n = 4 to 6 (4 to 6 replicates). Results are expressed as mean percentages compared to the untreated control standardized to 100%. Statistical analysis was performed against the untreated control after verifying the normality of the distribution of values ​​(Shapiro-Wilk test) using the One Way ANOVA test (Dunnett's method). The significance level was set at 5%.

[0132] Compared to the untreated control, fibulin-5 and MFAP4 levels were increased by +22% and +38%, respectively, at a dose of 0.1 pg / ml of the peptide. The peptide according to the invention therefore increases the levels of fibulin-5 and MFAP4 synthesized by fibroblasts and thus contributes to the organization of elastic microfibrils.

[0133] Example 6. Evaluation of the properties of the peptide according to the invention on dermal density

[0134] A population of 28 women aged 45 to 65 years with Fitzpatrick skin type I, II, or III applied a cream emulsion to half of their face. The cream contained a final concentration by weight of the peptide (Example LC) of 0.1% (w / w) relative to the total weight of the formulation (Formulation Example 7.c), or no peptide replaced by water (placebo). The application was repeated twice daily for two months. Total dermal density was assessed on the cheeks using a device called a DUB Skin Scanner, which uses the principle of ultrasound echogenicity to measure the fiber network density of the dermal extracellular matrix. The efficacy of the composition containing the peptide (Example LC) was compared with that of the placebo emulsion.

[0135] Total dermal density measurements were taken at baseline (D0), after one month of application (D28), and finally after two months of application (D56). The peptide's efficacy on total dermal density was determined by comparing the percentage change after 28 and 56 days of application relative to D0 of the peptide-containing formulation (as shown in the LC example) with that of the placebo formulation. Values ​​are expressed as the mean across the analyzed volunteers. Statistical analysis was performed after verifying the normality of the distribution of values ​​(Shapiro-Wilk test) using Student's t-test. The significance level was set at 5% (p<0.05).

[0136] After 28 and 56 days, the total dermal density of the formulation containing the peptide (according to example LC) was increased by 4.9% (p>0.05) and 12.9% (p<0.05), respectively. The peptide according to the invention therefore increases the density of the fiber network in the ECM.

[0137] The proportions are expressed as % by total weight and the names in capital letters correspond to the INCI names of the ingredients.

[0138] 7,a)

[0139] [Tables2] INCI name Quantity (% by total weight) Water qs 100.00 Vegetable oil 4.00 Coco-Caprylate / Caprate 4.00 Dycaprylyl Ether 4.00 Sodium Stearoyl Glutamate 1.00 Sodium Polyacrylate 1.00 Hydrogenated Vegetable Glycerides 1.00 Preservative qs peptide according to the invention, for example 0.50

[0140] 7,b)

[0141] [Tables3] INCI Name Quantity (% by total weight) Water qs 100 Xanthan Gum 10.00 Coco-Caprylate / Caprate 5.00 Cetearyl Alcohol 3.50 Glycerin 3.00 Caprylic / Capric Triglyceride 3.00 Pentylene Glycol 2.00 Sodium Polyacrylate (and) Dicaprylyl Carbonate (and) Polyglyceryl-3 Caprate 1.20 Sucrose Polystearate (and) Cetyl Palmitate 1.00 Phenoxyethanol (and) Ethylhexylglycerin 0.50 Dimethicone 0.50 Disodium Cetearyl Sulfosuccinate 0.20 Peptide according to the invention as shown in Example 1b 0.0001

[0142] 7,c)

[0143] [Tables4] INCI Name Quantity (% by total weight) water 80.27 Coco-Caprylate 6.50 Dimethyl Isosorbide 2.00 Polysorbate-20 2.00 Polyglyceryl-2 Dipolyhydroxystearate 2.00 Glycerin 2.00 Sodium Lauryl Glucose Carboxylate (and) Lauryl Glucoside 1.75 Sodium Polyacrylate 1.50 Phenoxyethanol (and) Ethylhexylglycerin 1.1 Maltodextrin 0.60 Peptide according to the invention according to the example 0.10 Xanthan Gum 0.10 Sodium Stearoyl Glutamate 0.08

[0144] The peptide used in the context of the invention is that shown in Table 5 below:

[0145] [Tables5] Sequence number: SEQIDN°1 TVFDGVLRPGQL

Claims

Demands

1. Non-therapeutic cosmetic use of SEQ ID No. 1 peptide and / or its homologs, derivatives and / or salts to improve the extracellular matrix fibers of the dermis of healthy skin and / or healthy mucous membranes to - increase the density and / or flexibility of healthy skin and / or healthy mucous membranes and / or - improve the anchoring of hair, preferably hair.

2. Use according to claim 1 to improve the structure and / or organization and / or density of collagen and / or elastic fibers of the extracellular matrix of the dermis.

3. Use according to any one of the preceding claims to prevent and / or slow down the loss of skin appendages, preferably hair.

4. Use according to any of the preceding claims to increase the amount of fibrillin 1 in the ECM fibers of the healthy dermis and / or to increase the amount of EMILIN-1 in the ECM fibers of the healthy dermis and / or to increase the amount of collagen fibers in the extracellular matrix of the healthy dermis, to increase the synthesis of fibulin 5 and / or MFAP4 by fibroblasts of the healthy dermis.

5. Use according to any one of the preceding claims wherein the peptide is in the form of the pure SEQ ID No. 1 peptide at least 50% (w / w).

6. Use according to any one of claims 1 to 5 wherein the SEQ ID No. 1 peptide is in the form of a peptide hydrolysate of Oryza saliva, in particular enriched in SEQ ID No. 1 peptide, preferably in liquid form.

7. Use according to any one of claims 1 to 5 wherein the SEQ ID No. 1 peptide is in the form of a microorganism fermentate producing, preferably selected from recombinant yeasts of the genus Pichia or Saccharomyces.

8. Use according to the preceding claim wherein the yeast is Saccharomyces cerevisiae.

9. Use according to any one of the preceding claims, characterized in that the SEQ ID No. 1 peptide and / or its homologs, derivatives and / or salts are in the form of a cosmetic composition comprising it and further advantageously comprising a suitable cosmetic vehicle.

10. Use according to claim 9, characterized in that the SEQ ID No. 1 peptide is present in the cosmetic composition in a content of between 1x10 6% to 10% by weight, preferably from 1x10 5% to 0.1% by weight, more advantageously from 1x10 4% to 0.01% by weight of dry matter relative to the total weight of the composition.

11. Use according to any one of the preceding claims characterized in that it is a topical use

12. Use according to any one of the preceding claims characterized in that it is intended for the care of sensitive and / or sensitized skin and / or mucous membranes.

13. A non-therapeutic cosmetic care and / or treatment method characterized in that it comprises the topical application to at least one area of ​​healthy skin, advantageously of a healthy scalp, and / or of healthy mucous membranes, of SEQ ID No. 1 peptide and / or its homologs, salts and / or derivatives, to improve the extracellular matrix fibers of the dermis of healthy skin and / or healthy mucous membranes, in particular the structure and / or organization and / or density of the collagen and / or elastic fibers of the extracellular matrix of the dermis, to - increase the density and / or flexibility of healthy skin and / or healthy mucous membranes and / or - improve the anchoring of the epidermis, preferably of hair.

14. Cosmetic care process according to claim 13 wherein the SEQ ID No. 1 peptide and / or its homologues and / or salts and / or derivatives is as defined in any one of claims 5 to 10.

15. A method according to any one of claims 13 or 14 wherein the skin and / or mucous membrane area is selected from the scalp, face, hands, arms, décolletage, legs, neck, back, shoulders, abdomen, wrists, forearms, ankles, thighs, nape of the neck, joint folds and / or armpits, labial mucous membranes.

16. SEQ ID No. 1 peptide and / or its homologs and / or salts and / or derivatives for its use in the treatment and / or prevention of a pathology associated with decreased ECM fibers and / or a decrease in Fibrillin-1 content and / or EMILIN-1 content and / or a decrease in the amount of collagen fibers and / or any combination thereof, said pathology being solar elastosis.

17. SEQ ID No. 1 peptide for its use according to claim 16 characterized in that the SEQ ID No. 1 peptide and / or its homologs is as defined in any one of claims 5 to 8.

18. SEQ ID No. 1 peptide for use according to any one of claims 16 or 17, characterized in that the peptide is present in a pharmaceutical composition in a content of between 1x10 6% and 10% by weight, preferably from 1x10 5% to 0.1% by weight, more advantageously from 1x10 4% to 0.01% by weight, of dry matter relative to the total weight of the composition, said composition further advantageously comprising a suitable pharmaceutical vehicle.

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