Active ingredient comprising oligoglucans derived from the cytosolic fraction of Saccharomyces cerevisiae and its cosmetic uses

Abstract

The present invention relates to an active principle comprising oligoglucans, obtained from the cytosolic fraction of a yeast of the species Saccharomyces cerevisiae, a cosmetic composition comprising it and its cosmetic uses, in particular to improve skin regeneration and combat its aging.

Description

Title of the invention: Active ingredient comprising oligoglucans derived from the cytosolic fraction of Saccharomyces cerevisiae and its cosmetic uses. Technical field

[0001] The invention relates to an active principle comprising oligoglucans, obtained from the cytosolic fraction of a yeast of the species Saccharomyces cerevisiae, a cosmetic composition comprising it and its cosmetic uses, in particular to improve skin regeneration and combat skin aging.

[0002] State of the art

[0003] The regeneration process, for example cutaneous, is a dynamic and complex process involving many cellular actors, during which new tissue develops to restore previously damaged areas and thus ensure its healing.

[0004] At the level of the skin tissue, three systems are involved in the process of tissue regeneration: the cutaneous, vascular and immune systems.

[0005] Skin tissue is now recognized as a complex organ including fibroblasts which play the pivotal role between these three systems to act on epidermal renewal, matrix remodeling, but also on vascularization, thus allowing the regeneration of damaged tissue.

[0006] At the level of the cutaneous system, epidermal renewal depends on keratinocytes, which cease adhering to the basement membrane to migrate, proliferate, and differentiate into functional keratinocytes. In the dermis, matrix remodeling is carried out by fibroblasts, after they have migrated and differentiated into myofibroblasts.

[0007] The vascular system enables, among other things, the transport of nutrients, mediators, and cells, which are essential for skin regeneration. To do this, endothelial cells migrate to group together cohesively, in order to form an organized and regular vascular network.

[0008] Finally, at the level of the immune system, there is a population of macrophages called pro-regenerative capable of interacting with fibroblasts, allowing to stimulate matrix remodeling and promote vascularization.

[0009] During the regeneration process, fibroblasts secrete growth factors that act on these systems by activating cell proliferation, migration, and differentiation pathways. These factors act in a paracrine manner. on the metabolism of surrounding fibroblasts but also on the metabolism of other cell types present in skin tissue.

[0010] However, during aging, cellular metabolism slows down. The ability of fibroblasts to secrete growth factors to interact with their environment decreases. The endogenous regeneration process then slows down, and the successive damages our skin sustains are no longer properly repaired. The quality of epidermal renewal decreases, the dermal matrix sags, and the vascular network becomes disorganized.

[0011] The first signs of skin aging then appear. The complexion becomes dull, loses its evenness, the skin is more prone to redness, and wrinkles become more pronounced. However, radiance and wrinkles are indicators of skin health. A dull complexion gives a tired appearance and can accentuate the signs of aging, which can have a significant impact on self-esteem and the image one projects.

[0012] Many cosmetic products aim to combat the signs of skin aging, for example to fight wrinkles, improve skin hydration, or enhance complexion radiance. However, these cosmetic ingredients act on one of the systems and do not aim for a comprehensive action on the entire regeneration process.

[0013] The exogenous supply of growth factors for tissue regeneration has certainly demonstrated its effectiveness. However, this solution relies on regenerative medicine techniques that are considered highly invasive. Therefore, it cannot be incorporated into a beauty routine.

[0014] Another rejuvenation technique called PRP (platelet-rich plasma) involves an intradermal injection of an autologous concentrate of growth factors present in PRP, previously obtained from a blood sample taken from the patient. Again, this technique is controversial due to its invasive nature and its effectiveness, which is limited by the patient's initial PRP concentration.

[0015] Facilitating skin regeneration processes while minimizing the impact on the patient remains a major challenge for the dermatological and cosmetic industries.

[0016] The inventor thus studied the metabolism of aged fibroblasts, as well as the metabolism of keratinocytes, endothelial cells and macrophages in the presence of the secretome of aged fibroblasts and observed: - a slowing of fibroblast migration and their differentiation into myofibroblasts, leading to a reduction in the synthesis of the collagen I network; - a decrease in keratinocyte migration, a key step in epidermal renewal; - a decrease in the pro-regenerative immune population, involved in matrix remodeling; - a reduction in endothelial cell metabolism, resulting in defective formation of the vascular network.

[0017] The metabolism of the cutaneous, immune and vascular systems is therefore reduced, not allowing optimal regeneration.

[0018] For this purpose, the inventor has identified a specific active ingredient obtained from a cytosolic fraction of the yeast Saccharomyces cerevisiae which can act on the entire process of skin regeneration.

[0019] Admittedly, cosmetic active ingredients obtained from Saccharomyces cerevisiae are already known. One example is an anti-aging solution containing a culture lysate of S. cerevisiae that stimulates skin cell biosynthesis (US5019391).

[0020] Other anti-aging and moisturizing cosmetic solutions are based on yeast hydrolysates, particularly yeast hydrolysates of S. cerevisiae. One example is a cosmetic composition comprising mannoproteins derived from the cell walls of yeasts such as Saccharomyces, as described in EP2567688. This extract is obtained from a chemical, enzymatic, or physicochemical hydrolysis of the yeast cell walls.

[0021] Application WO 2009 / 101503 also discloses the use of S. cerevisiae yeast hydrolysate consisting of proteins from the cell wall of hydrolyzed yeasts with a molecular weight between 1 and 5 kDa.

[0022] Thus, the solutions already known target in particular a hydrolysate of S. cerevisiae and more specifically a hydrolysate obtained from the cell wall of the S. cerevisiae yeast. This hydrolysate is rich in proteins, known to combat skin aging.

[0023] There is therefore a need for a non-invasive, natural cosmetic alternative that also acts on the cutaneous, vascular and immune systems simultaneously in order to stimulate the skin's endogenous capacities to improve its regenerative capacities. Summary of the invention

[0024] This is the objective of the present invention which, for this purpose, proposes a new active principle obtained from the cytosolic fraction of the yeast Saccharomyces cerevisiae.

[0025] A yeast is a eukaryotic microorganism, composed of a cell wall surrounding the plasma membrane. This wall consists of an outer layer of mannoproteins associated with glucans and an inner layer of glucans associated with chitins. The cytoplasmic membrane is composed mainly of phospholipids and protein complexes. The cell's contents, including the nucleus and cytoplasmic organelles, are contained within a liquid phase called the cytosol.

[0026] The yeast Saccharomyces cerevisiae is a well-known unicellular eukaryote, occurring as isolated, ovoid to rounded cells. It has been used since antiquity for the production of fermented beverages. However, it was only in the mid-19th century that it was isolated and identified as a replacement for traditional bakers' sourdough starter through its use in a fermentation process that was more reliable and faster than sourdough.

[0027] Uses as a medicinal product for certain gastrointestinal diseases have also been reported, as well as as a cosmetic active ingredient, as previously mentioned.

[0028] However, the inventor has surprisingly discovered the effects associated with an extract obtained specifically from the cytosolic fraction of a Saccharomyces cerevisiae yeast. Said extract comprises oligoglucans with a molar mass between 2 and 5 kDa.

[0029] Thus, the invention relates to an active principle comprising at least one extract of Saccharomyces cerevisiae, said extract corresponding to the cytosolic fraction comprising oligoglucans with a molar mass between 2 and 5 kDa.

[0030] Preferably, said extract consists of the cytosolic fraction of a Saccharomyces cerevisiae yeast. More preferably, the extract is a cytosolic fraction obtained by a mechanical lysate of Saccharomyces cerevisiae.

[0031] The invention also relates to a method for preparing the active ingredient according to the invention, comprising in particular solubilization of Saccharomyces cerevisiae in water by mechanical homogenization, i.e. a mechanical lysate allowing the yeast membranes to be broken down to extract the cytosolic fraction, separation of the soluble and insoluble phases, ultrafiltration followed by recovery of the filtrate and molecular sorting by ion chromatography.

[0032] The active ingredient according to the invention is of natural origin and advantageously improves skin regeneration. To achieve this, the active ingredient promotes the interconnectivity of the cutaneous, vascular, and immune systems. It acts on these three systems, thereby combating skin aging by providing a comprehensive solution. Thus, the active ingredient reduces dark circles and wrinkles and improves skin radiance. It therefore addresses the problems identified in the prior art.

[0033] The active ingredient according to the invention can be used for cosmetic applications, preferably when incorporated into a cosmetic composition, preferably a composition in a form suitable for topical application.

[0034] The invention therefore also relates to a cosmetic use of a composition comprising at least 0.1% by weight of the active ingredient according to the invention, in particular to improve skin regeneration. Preferably, the invention improves the interconnectivity of the cutaneous, vascular, and immune systems.

[0035] Other features and advantages will become apparent from the detailed description of the invention and the examples that will follow. Brief description of the Figures

[0036] [Fig. 1] represents the effect of the active ingredient according to the invention on the ability to skin regeneration, after 14 days of twice-daily application.

[0037] [Fig.2] represents the effect of the active ingredient according to the invention on the properties skin biomechanics after 21 and 42 days of twice-daily treatment. Detailed description of the invention

[0038] Definition

[0039] For the purposes of this invention, "cosmetic active ingredient" means an extract comprising at least one molecule, preferably a group of molecules, having a cosmetic effect on the skin. Preferably, the cosmetic effect on the skin is an anti-aging effect, in particular a skin-regenerating effect.

[0040] For the purposes of this invention, the term "regenerative effect" refers to the capacity of living organisms to repair themselves, and in particular the ability of the skin to regenerate in response to an injury (burn, scratch, cut) or to natural degradation. This includes a global effect that acts on the cutaneous, vascular, and immune systems by improving the interconnectivity of these systems.

[0041] For the purposes of this invention, "interconnectivity" refers to the exchanges between the different systems (cutaneous, vascular, and immune) within the skin tissue. In the context of this invention, the fibroblast plays a pivotal role in regeneration by interconnecting the three systems via the secretion of a pool of growth factors.

[0042] The term "system" refers to a set of interdependent components. In the context of the invention, these components include the cutaneous, vascular, and immune systems. The vascular system consists of arterioles, capillaries, and venules. The cutaneous system is the surface in contact with the external environment and comprises the epidermis, dermis, and hypodermis. The immune system consists of several types of immune cells, including dendritic cells, T lymphocytes, macrophages, and mast cells.

[0043] For the purposes of this invention, "mechanical lysate" means the product resulting from mechanical lysis, that is, the destruction of the membrane of a biological cell by a physical agent such as a mechanical means, as opposed to a chemical agent or biological factors such as temperature, an enzyme, or the natural autolysis of the cell.

[0044] For the purposes of this invention, "mechanical lysate of Saccharomyces cerevisiae" means any active ingredient derived from the yeast Saccharomyces cerevisiae, obtained by a process comprising at least one solubilization step of Saccharomyces cerevisiae consisting of mechanical homogenization. Mechanical lysis, that is, the destruction of the membrane of a biological cell, is carried out by a physical technique such as a mechanical means, as opposed to a chemical or biological agent such as an enzyme, temperature, or even the natural autolysis of the cell. The term "mechanical lysate" excludes molecules produced solely by fermentation of Saccharomyces cerevisiae, as well as those resulting from enzymatic or chemical hydrolysis.

[0045] For the purposes of this invention, "oligoglucan" means a glucan fraction. Glucans and oligoglucans are polysaccharides composed exclusively of glucose. These glucose molecules can be linear or branched. In particular, an oligoglucan is a small glucose oligomer, i.e., less than or equal to 5 kDa.

[0046] Active ingredient according to the invention

[0047] The present invention therefore relates to a cosmetic active ingredient comprising at least one yeast extract of the species Saccharomyces cerevisiae, particularly useful for improving skin regeneration.

[0048] The active principle according to the invention comprises at least one extract of Saccharomyces cerevisiae, said extract corresponding to the cytosolic fraction comprising oligoglucans with a molar mass between 2 and 5 kDa.

[0049] Thus, said extract excludes any other part or element of the yeast, such as the yeast cell wall or membranes. Preferably, said extract consists of the cytosolic fraction of the yeast Saccharomyces cerevisiae. Advantageously, the extract is a mechanical lysate of Saccharomyces cerevisiae.

[0050] According to a particularly preferred object, said extract is capable of being obtained by a process comprising the following steps: a. solubilization of at least 50 g / L of Saccharomyces cerevisiae in water, by mechanical homogenization, b. separation of soluble and insoluble phases, c. ultrafiltration and recovery of the filtrate, and d. molecular sorting by ion chromatography.

[0051] Preferably, the extract is obtained from the soluble phase.

[0052] The extract constituting the cosmetic active ingredient according to the invention has been characterized by the inventor. It comprises oligoglucans which preferably represent between 10% and 35% by weight of dry matter of the extract.

[0053] Preferably, the extract according to the invention, being composed of the cytosolic fraction of the yeast, does not include membranes and consequently contains little mannose. Even more preferably, the extract according to the invention comprises a glucose / mannose ratio greater than 60%.

[0054] Preferably, the extract according to the invention also comprises a protein fraction. The distribution and quantity of the protein fraction can be determined by the total nitrogen assay according to the Kjeldahl method (reference: Official method of analysis of the AOC, 12th ed. W. Horwitz, ED, New York, 15-60, 1975). Preferably, the extract comprises between 45 and 55% peptides by dry weight of the extract according to the Kjeldahl method, of which 96% are peptides having a molecular mass of less than 2,000 Da.

[0055] The inventor also determined the ash content. The ash content can be determined by weighing the residues from the incineration of samples of the active ingredient according to the invention at 550°C in an electric muffle furnace. Preferably, the active ingredient according to the invention has an ash content of between 10% and 22% by weight of the dry matter of the active ingredient.

[0056] In addition, the extract according to the invention also comprises between 0 and 31% by dry weight of other cellular metabolites present in the cytosol such as organic acids, DNA, etc.

[0057] The cosmetic active ingredient according to the invention can be in liquid form, solid form or film form.

[0058] When the active ingredient according to the invention is in liquid form, it consists exclusively of the extract of the cytosolic fraction of the yeast Saccharomyces cerevisiae, possibly accompanied by a stabilizer and / or preservation system.

[0059] The extract in liquid form is preferably a clear aqueous solution with a characteristic odor and a light yellow color. However, it may be more colored and / or be decolorized by any process known to those skilled in the art.

[0060] The dry matter content can be determined by weighing the residues obtained from drying the extract according to the invention at 105°C in an oven until a constant weight is obtained. Preferably, the extract according to the invention in liquid form has a dry matter content of: 10 g / L to 50 g / L, even more preferably of 15 g / L to 35 g / L.

[0061] The active ingredient according to the invention can also be presented in the form of a film. In this case, the extract of the cytosolic fraction of Saccharomyces cerevisiae preferably represents at least 0.1% by weight of the film.

[0062] When in film form, the active ingredient comprises: a. at least the lysate of the cytosolic fraction of Saccharomyces cerevisiae according to the invention, and b. at least one mineral filler, and / or c. at least one polymer of natural origin, and / or d. at least one plasticizer, and / or e. at least one surfactant.

[0063] The polymer of natural origin can be chosen from: Pectin, Tamarind gum, Alginate, Pullulan, Psyllium, Xanthan gum, Guar gum, Tara gum, Carob gum, Agar, Gum arabic, Gellan gum, Dextran, Carrageenan, Cellulose, Konjac and Chitosan.

[0064] The plasticizer can be chosen from: Glycerol, Sorbitol, Sucrose, Erythritol, Urea, Propylene glycol and Butylene glycol.

[0065] The mineral filler may be selected from: calcium carbonate, green clay, kaolin, perlite, talc, magnesium silicate, mica, diatomaceous earth, silica, calcium sulfate, calcium chloride, potassium chloride, iron oxide, and zinc oxide. The active ingredient may further include a pigment for coloring the film.

[0066] When the active ingredient according to the invention is in solid form, it preferably consists of the extract of the cytosolic fraction of Saccharomyces cerevisiae yeast as previously described and a carrier selected from maltodextrin, acacia gum, soy lecithin, or isomalt. In a particularly suitable embodiment, the extract represents at least 10% by weight of the active ingredient and the carrier at most 90% by weight of the active ingredient.

[0067] In the case of a solid form in which the active ingredient is associated with a support, the sugar, protein fraction and ash contents in the active ingredient are modified, the support generally being mainly composed of sugars.

[0068] Extraction process

[0069] The extract constituting or contained in the active principle according to the invention can be obtained by any process comprising at least one step of extraction of the cytosolic fraction of the yeast of Saccharomyces cerevisiae, in particular a step of mechanical lysis.

[0070] According to a particularly suitable embodiment, the active ingredient according to the invention is obtained by carrying out the following steps: a. solubilization of at least 50 g / L of Saccharomyces cerevisiae in water, by mechanical homogenization, b. separation of the soluble and insoluble phases, c. ultrafiltration and filtrate recovery, and d. Molecular sorting by ion chromatography.

[0071] Mechanical homogenization is carried out by any means known to a person skilled in the art, for example by mechanical grinding using a homogenizer.

[0072] The separation of the soluble and insoluble phases is carried out by any means known to those skilled in the art, for example by centrifugation, filtration or decantation. Preferably, the separation of the soluble and insoluble phases is carried out by centrifugation, thus allowing the recovery of the soluble phase containing, among other things, the soluble sugar and protein fractions.

[0073] The process includes an ultrafiltration step after recovery of the soluble phase to perform molecular sorting. This ultrafiltration step purifies the recovered soluble phase. It is carried out to remove high molecular weight molecules (enzymes and polymers, cell wall components, membranes, etc.), that is, molecules with a molecular weight greater than 1000 kDa.

[0074] The lysate obtained at this stage can optionally be further concentrated and / or purified, preferably by successive ultrafiltration steps through filters with different cut-off thresholds, retaining the filtrates at each step and / or by a chromatographic method.

[0075] The product obtained after lysis and ultrafiltration, before or after concentration and sterilizing filtration, is a lysate of the cytosolic fraction of Saccharomyces cerevisiae, and constitutes the first form of the active principle according to the invention, then being in liquid form.

[0076] The lysate obtained can then be dried and combined with a support to obtain a solid form. This phase can be carried out by implementing the following steps: a. an atomization support, preferably maltodextrin, is added to the product in liquid form, at most 90% (by mass / volume), b. this solution is then concentrated under vacuum; c. Any bacteria present are eliminated by heat treatment or filtration; d. Atomization allows us to obtain a powder.

[0077] The steps of the processes described above, taken individually, are usual in the field of extracting active ingredients from natural raw materials and a person skilled in the art is able to adjust the parameters on the basis of their general knowledge.

[0078] Composition

[0079] The active ingredient according to the invention can be incorporated into a cosmetic composition, in particular a composition comprising at least 0.1% by weight of said active ingredient.

[0080] Said cosmetic composition according to the invention preferably comprises a physiologically acceptable medium, preferably a cosmetically acceptable medium, that is to say, one which does not cause any discomfort for the user such as redness, tightness or tingling.

[0081] The composition can be presented in different galenic forms, adapted for topical application to the skin.

[0082] These compositions may be in the form of oil-in-water emulsions, water-in-oil emulsions, multiple emulsions (Water / Oil / Water or Oil / Water / Oil), which may be microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersions, aqueous gels, powders, or foundation. They may be more or less fluid and have the appearance of creams, emulsions, gels, masks, or any other aspect of cosmetics for the care of healthy skin.

[0083] These may be compositions comprising at least 0.1% of the liquid active ingredient according to the invention, preferably between 0.5 and 10% or comprising at least 0.01% of a powdered active ingredient according to the invention.

[0084] The compositions according to the invention may contain as an adjuvant at least one compound selected from: a. oils, which can be chosen in particular from among silicone oils, linear or cyclic, volatile or non-volatile, b. waxes, such as ozokerite, polyethylene wax, beeswax or camauba wax, c. silicone elastomers, d. surfactants, preferably emulsifiers, whether non-ionic, anionic, cationic or amphoteric, e. co-surfactants, such as linear fatty alcohols, f. thickeners and / or gelling agents, g. humectants, such as polyols like glycerin, h. colorants, preservatives, fillers, i. tensors, j. the sequestrators, k. perfumes, 1. and their mixtures, without this list being exhaustive.

[0085] Examples of such adjuvants are cited in particular in the CTFA Dictionary (International Cosmetic Ingredient Dictionary and Handbook published by the Personal Care Product Council).

[0086] Of course, a person skilled in the art will take care to choose any additional compounds, active or inactive, and their quantity, in such a way that the advantageous properties of the mixture are not, or substantially not, altered by the envisaged addition.

[0087] These compositions are intended for use on healthy skin, particularly mature skin, especially to improve skin regeneration and achieve an overall anti-aging effect, preferably an anti-wrinkle effect and / or to improve the radiance of the complexion and / or reduce dark circles.

[0088] The invention therefore also relates to a cosmetic skin treatment process to improve skin regeneration, in particular to improve the radiance of the complexion and / or reduce dark circles and / or reduce wrinkles, which consists of the topical application on the skin of a healthy person of such an active ingredient according to the invention or of such a composition according to the invention.

[0089] Use

[0090] The active principle according to the invention or the composition according to the invention has characteristics which allow its use in cosmetics and in particular to limit the natural effects related to skin aging and thus improve skin regeneration and obtain an overall anti-aging effect, preferably reducing wrinkles and / or reducing dark circles and / or improving the radiance of the complexion.

[0091] To achieve this, the active ingredient or composition according to the invention improves the interconnectivity of the cutaneous, vascular and immune systems. Preferably, the active ingredient or composition according to the invention increases the synthesis and secretion of the growth factors KGF, EGF, IGF, FGF, PDGF, TGF-3 and VEGF.

[0092] In the cutaneous system, specifically in the epidermis: KGF (keratinocyte growth factor) is known to stimulate keratinocyte migration, an essential step in epidermal renewal. Similarly, EGF (epidermal growth factor) promotes the proliferation of epithelial cells. Finally, IGF (insulin growth factor) promotes keratinocyte proliferation, migration, and differentiation. In the dermis, matrix remodeling is induced by four main growth factors. Thus, IGF has a mitogenic effect on fibroblasts; FGF-2 or bFGF, a member of the FGF (fibroblast growth factor) family, promotes fibroblast proliferation and migration; and PDGF (platelet-derived growth factor) also stimulates fibroblast survival and collagen synthesis. Finally, TGF-[3 (transforming growth factor) stimulates fibroblast proliferation and induces their differentiation into myofibroblasts.

[0093] At the level of the immune system, TGF-[3] released by fibroblasts is able to activate resident macrophages into pro-regenerative macrophages. They then secrete numerous growth factors, such as TGF-[3].

[0094] At the level of the vascular network: VEGF (vascular endothelial growth factor) promotes the formation of new capillaries by endothelial cells.

[0095] The active principle according to the invention, by increasing the synthesis and secretion of KGF, EGF, IGF, FGF, PDGF, TGF-[3 and VEGF, thus makes it possible to respectively stimulate keratinocyte migration, the proliferation of epithelial cells, keratinocyte differentiation, promote the proliferation and migration of fibroblasts, stimulate fibroblast survival and collagen synthesis, stimulate fibroblast proliferation and induce their differentiation into myofibroblasts and thus activate resident macrophages into pro-regenerative macrophages.

[0096] Therefore, the active ingredient according to the invention acts on: • The cutaneous system: by significantly improving fibroblast migration (+97%), their differentiation into myofibroblasts, essential players in skin regeneration (α-SMA increased by 40%), and by promoting the synthesis of the collagen 1 network (+53%), the active ingredient stimulates matrix renewal. This translates in vivo into an improvement in dermal structure: matrix quality is increased by 32% and fiber quality by 20%, compared to placebo. In the epidermis, the active ingredient also stimulates keratinocyte migration (+99%), essential for epidermal dynamics. • the immune system: by promoting the pro-regenerative population (+132%), capable of interacting back with fibroblasts, it helps to promote matrix remodeling. • the vascular system: by restoring the migration of endothelial cells and their cohesion (+93% and +77% respectively), the active ingredient promotes the formation of the vascular network (+55%).

[0097] By restoring all growth factors, the active ingredient according to the invention thus exhibits a comprehensive anti-aging action. Consequently, wrinkles and dark circles are significantly reduced, the microvascular network becomes homogeneous again, the skin is firmer, and the complexion's radiance is revived. It is particularly suitable for healthy, mature skin.

[0098] The invention is now illustrated by non-limiting examples of compositions according to the invention and by test results. Examples

[0099] Example 1 - Active ingredient according to the invention

[0100] The active ingredient is obtained by carrying out the following steps: a. Solubilization by mechanical homogenization (lysis) of Saccharomyces cerevisiae yeast, b. Separation of the soluble / insoluble phases, c. Ultrafiltration, d. Filtrate recovery, e. Molecular sorting by ion chromatography, intended to select the molecules, f. Bleaching and Deodorizing, and g. Filtration and sterilizing filtration.

[0101] The extract constituting the active principle according to the invention consists of the cytosolic fraction of S. cerevisiae yeasts, the membranes are eliminated and virtually no mannose remains in said extract (the mannose level of the glycosidic fraction is less than 2%).

[0102] The active ingredient obtained has the following analytical characteristics: a. a dry matter content: 24.4 g / L, b. a protein fraction content according to the Kjeldahl method: 12.4 g / L (50.8%), c. a total sugar content: 4 g / L (16.4%), d. an ash content: 4.3 g / L (17.6%), e. cellular metabolites, organic acids, DNA, etc. (15.20%).

[0103] The glycosidic fraction of the active ingredient obtained is more particularly composed of 10% monosaccharides with a molar mass of less than 180 Da and 90% oligosaccharides with a molar mass between 360 Da and 5,000 Da. More specifically, the glycosidic fraction is composed of approximately 0.5% glucosamine, 1.7% galactose, 1.4% mannose, and 96.4% glucose, resulting in a glucose / mannose ratio of 68.9%.

[0104] The sugars present in the active ingredient are in the form of oligoglucans, preferably of size between 2 and 5 kDa.

[0105] Example 2 - Active ingredient not part of the invention

[0106] The active ingredient is obtained by carrying out the following steps: a. Solubilization of Saccharomyces cerevisiae yeast in a basic aqueous medium at a concentration of at least 200g / L, b. Enzymatic hydrolysis, c. Separation of soluble and insoluble phases by centrifugation, decantation or filtration, d. Sterilizing filtration

[0107] The active ingredient obtained has the following analytical characteristics: a. a dry matter content: 54.6 g / L, b. a protein content according to the Lowry method: 23.2 g / L (42%), c. a total sugar content: 18.3 g / L (34%),

[0108] The active ingredient according to example 2 consists of 34% sugars, of which 5% are disaccharides and 95% are saccharides with a size greater than 1440 Da. And a glucose / mannose ratio of 14.0%.

[0109] Example 3 - Active ingredient not part of the invention

[0110] The active ingredient is obtained by carrying out the following steps: a. Solubilization of Saccharomyces cerevisiae yeast in a slightly basic aqueous medium at a concentration of at least 20%, possibly with the addition of an alcohol to promote yeast solubilization. b. hydrolysis of proteins in a basic medium, c. Agitation from 8 to 52 hours, d. First filtration to separate the soluble and insoluble phases, e. washing with water to purify the active fraction, f. successive concentrations and filtrations, and g. sterilizing filtration.

[0111] The active ingredient obtained has the following analytical characteristics: a. a dry matter content: 42.7 g / L, b. a protein content according to the Lowry method: 10.4 g / L (24%), c. a total sugar content: 25.7 g / L (60%),

[0112] The active principle according to example 3 is thus mainly composed of mannans.

[0113] Example 4 - Composition according to the invention

[0114] An example of a formulation comprising the active ingredient according to the invention in the form of an anti-aging gel-cream characterized by a fresh and creamy texture is shown in Table 1 below.

[0115] [Tables 1] Ingredients % Al Water qsp 100 A2 Butylene Glycol 3.00 Sclerotium Gum & Xanthan Gum 0.20 B Dimethicone 8.00 Dimethicone 8.00 Caprylyl Methicone 4.00 Polysorbate 80 2.00 Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolyme r 0.87 C Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolyme r & Squalane & Polysorbate 60 2.00 D Active principle according to the invention 2.50 E Preservative qs

[0116] The composition of Example 4 can in particular be obtained by the following process: a. Add A2 to Al while stirring and homogenize for 20 minutes, b. Add B then C to A while stirring vigorously. Shake for 20 minutes. c. Add D and E under moderate stirring.

[0117] The composition is then in the form of a thick, glossy, off-white cream gel.

[0118] Example 5 - Composition according to the invention

[0119] An example of a formulation comprising the active ingredient according to the invention in the form of a repairing day cream is shown in Table 2 below.

[0120] [Tables2] Ingredients % Al Water qs 100 Sodium Gluconate 0.20 Sodium Benzoate 0.30 A2 Pentylene Glycol 5.00 Sclerotium Gum & Seed Butter 1.50 Hydrogenated Vegetable Oil 0.75 Coco-Caprylate / Caprate 6.00 Triheptanoin 6.50 Caprylic / Capric Triglyceride 8.25 Helianthus Annuus (Sunflower) Seed Oil 4.00 Tocopherol & Helianthus Annuus (Sunflower) Seed Oil 0.10 C Active ingredient according to the invention 2.50 D Citric Acid Solution 10% qs pH

[0121] The composition of Example 5 can in particular be obtained by the following process: a. Add A2 to Al under moderate stirring, stir until a homogeneous gel is obtained and heat to 80°C, b. Place B under magnetic stirring and heat to 80°C. c. Emulsify B in A under shear agitation for 10 minutes, d. At 30°C, under moderate stirring, add C, and e. Adjust the pH to 5.0 - 5.5 with D.

[0122] The composition is then in the form of a soft and glossy off-white gelled cream.

[0123] Example 6 - Composition according to the invention

[0124] An example of a formulation comprising the active ingredient according to the invention in the form of a hybrid cream, half-skincare, half-foundation is shown in Table 3 below.

[0125] [Tables3] Ingredients % Al Water qs 100 Preservative qs A2 Glycerin 1.50 Sclerotium Gum & 4.00 Cl2-15 Alkyl Benzoate 3.00 Dimethicone 0.50 B2 Ethylhexyl Methoxycinnamate 3.00 Ethylhexyl Salicylate 3.00 C Polyacrylate-13 & Polyisobutene & Polysorbate 20 0.50 D Polyacrylate Crosspolymer-6 0.50 E Butylene Glycol 2.00 Alumina & CI 77891 (Titanium Dioxide) 1.50 Alumina & CI 77891 (Titanium Dioxide) 1.50 F Active principle according to the invention 2.50 G Butylene Glycol 2.00 Titanium Dioxide & Mannitol & CI 77492 (Iron Oxi des) & Hydrogenated Lecithin & Polymethyl Metha crylate & CI 77491 (Iron Oxides) & Aluminum Hyd roxide & CI 77499 (Iron Oxides) 3.00

[0126] The composition of example 6 can in particular be obtained by the following process: a. Add A2 to Al while stirring, stir until homogeneous and heat to 80°C. b. Place B1 under stirring and heat to 80°C. Add B2 to B1 just before the emulsion, c. Emulsify B in A under shear agitation for 10 minutes, d. At 50°C, under moderate stirring, add C then D, E and F, and e. At room temperature, add G under very gentle stirring.

[0127] The composition is then in the form of a soft, glossy, pale grey emulsion with grey particles in suspension.

[0128] Example 7 - Composition according to the invention

[0129] An example of a formulation comprising the active ingredient according to the invention in the form of a regenerating night cream is shown in Table 4 below.

[0130] [Tables4] Ingredients % Al Water qs 100 Preservative qs Sorbitol 70% 1 1.00 A2 Propylene Glycol 3.00 Cetearyl Ethylhexanoate 3.50 Octyldodecanol 3.50 Squalane 3.50 Pentaerythrityl Tetraisostearate 3.50 Diisopropyl Adipate 3.50 Dimethicone 1.50 Argania Spinosa Kernel Oil 2.00 Tocopherol & Helianthus Annuus (Sunflower) Seed Oil 0.05 C Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolym r & Polyisobutene & PEG-7 Trimethylolpropane Coconut Ether 1.00 D Active ingredient according to the invention 2.50

[0131] The composition of Example 7 can in particular be obtained by the following process: a. Add A2 to Al under moderate stirring, stir until a homogeneous gel is obtained and heat to 80°C, b. Place B under magnetic stirring and heat to 80°C. c. Emulsify B in A under shear agitation for 10 minutes, and d. At 50°C, under moderate stirring, add C then D.

[0132] The composition is then in the form of a flexible, glossy, pale beige emulsion. At 1 month, the pH is equal to 5.4 and the viscosity is equal to (C / 5rpm) 57,400 cP.

[0133] Evaluation of the cosmetic effect of the active ingredient according to the invention

[0134] Test 1 - Effect of the active ingredient according to the invention on its ability to restore the fibroblast regenerating complex.

[0135] The objective of this trial is to evaluate the impact of aging on the expression and synthesis of growth factors: TGF-[31, KGF, VEGF, FGF2, EGF, IGF1 and PDGFA present within the fibroblast regenerative complex.

[0136] For this study, the following parameters were analyzed on human fibroblasts from facial plastic surgery, young and old.

[0137] The expression of EGF, IGF1 and PDGFA was evaluated by qPCR and the synthesis of TGF-[31, KGF, VEGF and FGF2 was evaluated by ELIS A assay.

[0138] The operational protocol of the study is described below.

[0139] Young and old human fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0140] Then the cells are treated with the extracts at 0.25% and 1.00% (V / V). They are then incubated at 37°C in an atmosphere containing 5% CO2.

[0141] Fibroblasts are recovered and total RNA is extracted.

[0142] The RNAs were reverse-transcribed and the resulting complementary DNAs were analyzed by quantitative PCR. Internal reference control mRNAs were analyzed in parallel with EGF, IGF1, and PDGFA mRNAs.

[0143] Fluorescence incorporation (SYBR Green) is continuously measured using a thermocycler. Ct analysis (relative quantification) is performed using software.

[0144] The supernatants are recovered and stored at -80°C pending ELISA assays of TGF|31, KGF, VEGF, FGF2.

[0145] The objective of this study is to evaluate the effect of the examples according to the invention on the expression of the growth factors: EGF, IGF1 and PDGFA of the fibroblast regenerating complex.

[0146] The results are given in Table 5.

[0147] [Tables5] Ability to restore EGF expression (%) Ability to restore IGF-1 expression (%) Ability to restore PDGFA expression (%) Young fibroblasts Control Older fibroblasts Control -44 -42 -41 EXAMPLE 2 (HI) 1.00% -4 - INVENTION 0.25% +54 +50 +49 INVENTION 1.00% +98 +75 +76

[0148] Compared to young fibroblasts, aged fibroblasts have a reduced capacity to express: EGF by -44%; IGF-1 by -42%; PDGFA by -41%.

[0149] Aging leads to a significant depletion of growth factors in the fibroblast regenerative complex.

[0150] The active principle according to the invention, for its part, significantly restores the expression of the growth factors: EGF, IGF1 and PDGFA.

[0151] This test was also carried out in order to evaluate the effect of the active ingredient according to the invention on the synthesis of growth factors: TGF[31, VEGF, KGF and FGF2 of the fibroblast regenerating complex.

[0152] The results are given in Table 6.

[0153] [Tableauxô] Ability to restore TGF PI synthesis (%) Ability to restore VEGF synthesis (%) Ability to restore KGF synthesis (%) Ability to restore FGF2 synthesis (%) Young fibroblasts Control Older fibroblasts Control -19 -36 -44 -29 EXAMPLE 2 (HI) 0.50% - - 0 - EXAMPLE 2 (HI) 2.00% - - 0 0 EXAMPLE 3 (HI) 2.00% - - - 0 INVENTION 0.25% +85 +50 +25 +26 INVENTION 1.00% +135 +73 +48 +49

[0154] Compared to young fibroblasts, aged fibroblasts have a reduced capacity to synthesize: TGF-

[31] by -19%; VEGF by -36%; KGF by -44%; FGF2 by -29%.

[0155] Aging leads to a significant depletion of growth factors in the fibroblast regenerative complex.

[0156] The active principle according to the invention significantly restores the synthesis of growth factors, whereas the examples outside the invention do not show an effect on all the growth factors studied.

[0157] Tested at 1% on aged human fibroblasts, the active ingredient according to the invention significantly restores the expression or synthesis of growth factors: EGF by 98%, IGF1 by 75%, PDGFA by 76%, TGF[31 by 135%, KGF by 48%, VEGF by 92%, and FGF2 by 49%.

[0158] The active principle according to the invention thus makes it possible to enrich the fibroblastic regenerative complex of aged cells with growth factors.

[0159] Test 2 - Effect of the active ingredient according to the invention on fibroblast migration.

[0160] The objective of this study is to evaluate the effect of the active ingredient according to the invention on the migration of aged human fibroblasts.

[0161] In response to mechanical stress, cells move within the tissue to locate themselves at the pressure zone. At this point, contraction forces are then generated to maintain skin integrity. With age, the migratory capacity of cells is reduced.

[0162] This study was carried out by a time-lapse cell migration test on human fibroblasts from facial plastic surgery, young and old.

[0163] The effect is measured by assessing the ability of cells to recolonize an injured area.

[0164] Young and old human fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0165] The injured area is prepared after four days. The cells are treated with extracts at 0.25% and / or 1.00% and / or 2% (V / V). They are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0166] The visualization of fibroblast migration is carried out in "time-lapse" using a fully automated system which allows the migration of cells to be followed during culture.

[0167] The cell migration velocity within the wound is calculated using an internally developed image analysis script. The results are expressed in pm2 / min.

[0168] The results are given in Table 7.

[0169] [Tables7] Ability to restore migration velocity (%) Control (young fibroblasts) -27 INVENTION 0.25% (control) +54 INVENTION 1.00% (control) +97 EXAMPLE 2 (HI) 1% (control) +14 EXAMPLE 2 (HI) 2% (control) Negative

[0170] The migration velocity of aged human fibroblasts is significantly reduced by 27% compared to young human fibroblasts.

[0171] Tested at 1.0% on aged human fibroblasts, the active ingredient according to the invention significantly restores the migration rate by 97%. This contrasts with Example 2, which is not part of the invention and exhibits a slow migration rate.

[0172] The active principle according to the invention thus promotes a faster recolonization of the aged injured dermis, a key step in skin regeneration.

[0173] Test 3 - _ Effect of the active ingredient according to the invention on differentiation into myofibroblasts.

[0174] The objective of this study is to evaluate the effect of the active ingredient according to the invention on the synthesis of alpha-Smooth muscle actin (a-SMA), actin microfilaments characteristic of myofibroblasts, in an injured area.

[0175] In damaged tissues, fibroblasts are activated and differentiate into myofibroblasts, which contract and participate in healing by reducing the size of the wound and by secreting extracellular matrix proteins.

[0176] The study was carried out by immunocytofluorescence, on human fibroblasts from facial plastic surgery, young and old.

[0177] The operative protocol of the study is described below. Young and old human fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2 and cultured for several days.

[0178] The wound is created. The cells are treated with the active ingredient according to the invention at 0.25% and 1.00% (V / V) for 72 hours. They are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0179] Immunocytological labeling of a-SMA is carried out by fixation and permeabilization using a primary antibody and a secondary antibody coupled to a fluorophore.

[0180] Visualization is performed using a microscope coupled with a camera and an image analysis system.

[0181] The intensity of α-SMA labeling is proportional to the intensity of staining (fluorescence) present in the cytoplasm of fibroblasts. The greater the fluorescence, the higher the rate of α-SMA synthesis.

[0182] A quantitative analysis of the images was carried out using an internally developed image analysis script. The results are expressed in arbitrary units (AU).

[0183] The results are given in Table 8.

[0184] [Tables8] α-SMA Synthesis (xO2 UA) Capacity to stimulate α-SMA synthesis (%) Young fibroblasts Control 832 Older fibroblasts Control 205 -75% INVENTION 0.25% 242 +18% INVENTION 1.00% 286 +40%

[0185] The synthesis of a-SMA, a characteristic marker of myofibroblasts, is significantly reduced by 75% in an injured model in older cells compared to younger cells.

[0186] Tested at 1% on aged fibroblasts, the active ingredient according to the invention significantly stimulates the synthesis of a-SMA by 40%.

[0187] The active principle according to the invention thus enables the transformation of fibroblasts into myofibroblasts, key players in dermal regeneration.

[0188] Test 4 - Effect of the active ingredient according to the invention on the migration of keratinocytes.

[0189] The objective of this study is to evaluate the impact of the regenerating complex aged fibroblast treated with the active ingredient according to the invention on the migration of aged human keratinocytes.

[0190] In response to mechanical stress, cells move within the tissue to locate themselves at the pressure zone. At this point, contraction forces are then generated to maintain skin integrity. With age, the migratory capacity of cells is reduced.

[0191] This study was performed using a time-lapse cell migration assay on human keratinocytes from young and old donors. The effect is measured by assessing the cells' ability to recolonize an area of ​​injury.

[0192] Young and old human keratinocytes are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0193] The wound is created. The cells are treated with secretomes from young fibroblasts (SFJ) or old fibroblasts (SFA) in the presence or absence of the active ingredient according to the invention at 0.25% and 1.00% (V / V). The cells are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0194] The visualization of keratinocyte migration is carried out in "time-lapse" using a fully automated system which allows the migration of cells to be followed during culture.

[0195] The cell migration velocity within the wound is calculated using an internally developed image analysis script. The results are expressed in pm2 / min.

[0196] The results are given in Table 9.

[0197] [Tables9] Ability to restore migration velocity (%) Young keratinocytes SFJ Control SFJ INVENTION 1.00% +13 Aged Keratinocytes SFA Control -45 SFA INVENTION 0.25% +91 SFA INVENTION 1.00% +99 SFA EXAMPLE 2 (HI) 1.00% +39

[0198] Migration of aged human keratinocytes is significantly reduced by 45% compared to young human keratinocytes.

[0199] When tested on aged human keratinocytes, the secretome derived from aged human fibroblasts pretreated with 1.0% of the active ingredient according to the invention significantly restores cell migration velocity by 99%. Whereas in the presence of Example 2 outside the invention, cell migration velocity is restored by only 39%.

[0200] The active principle according to the invention thus accelerates keratinocyte migration, a key step in skin regeneration.

[0201] Test 5 - Effect of the active ingredient according to the invention on the immune system and macrophage polarization.

[0202] The objective of this study is to evaluate the impact of the aged fibroblastic regenerative complex treated with the active ingredient according to the invention on the polarization of pro-regenerative macrophages.

[0203] The immune system is involved in defenses against pathogens but also participates in tissue homeostasis. During wound healing, the regeneration phase is notably guided by a population of pro-regenerative macrophages with anti-inflammatory activity, capable, among other things, of stimulating matrix remodeling. With age, the activity of these macrophages decreases, with, in particular, a reduced reactivity to external stimuli leading to poorer differentiation and a decrease in the expression of certain surface receptors necessary for their function.

[0204] This study was conducted on human macrophages derived from monocytes extracted from human peripheral blood. Polarization into pro-regenerative macrophages was assessed by analyzing the synthesis of CD163, a specific marker of this population and contributing to the anti-inflammatory response, using flow cytometry. To ensure that no pro-inflammatory macrophage population was induced by the treatments, the synthesis of HLA-DR, a preferential marker of this population, was analyzed by flow cytometry and complemented by the analysis of TNFα synthesis using an ELISA assay.

[0205] The operative protocol of the study is described below. Human monocytes are isolated from peripheral blood mononuclear cells (PBMCs) and are seeded in a macrophage differentiation medium before being incubated at 37°C in an atmosphere containing 5% CO2.

[0206] The macrophages are then treated with secretomes from young (SFJ) or old (SFA) fibroblasts in the presence or absence of the active ingredient according to the invention at 0.25% and 1.00% (V / V). The cells are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0207] Macrophages are collected and labeled with CD163 and HLA-DR specific antibody conjugates. The cells are then fixed in a 1% paraformaldehyde solution before being analyzed on a flow cytometer equipped with analysis software.

[0208] The synthesis rate of CD 163 and HLA-DR is assessed via the median fluorescence associated with each of these markers and is expressed in arbitrary units (AU).

[0209] In parallel, the supernatants are collected and stored at -80°C until analysis by ELISA assay of TNFa.

[0210] The results are given in Table 10.

[0211] [TableauxlO] Pro-regenerative macrophages CD163 synthesis (UA) Ability to restore CD163 synthesis (%) SFJ Control 231,811 SFA Control 178,281 -23% SFA INVENTION 0.25% 248,847 +132

[0212] The secretome of young or old fibroblasts treated or not with the active ingredient according to the invention does not modify the rate of HLA-DR synthesis and does not induce TNFa synthesis.

[0213] The secretome derived from human fibroblasts promotes the generation of pro-regenerative macrophages without inducing pro-inflammatory macrophages.

[0214] The rate of synthesis of CD163, a specific receptor involved in the functionality of pro-regenerative macrophages, is decreased by 23% in the presence of secretome from aged human fibroblasts compared to secretome from young human fibroblasts.

[0215] The secretome derived from aged human fibroblasts pretreated with 0.25% of the active ingredient according to the invention restores the surface expression of CD163 by 132%.

[0216] The active principle according to the invention thus enhances the functionality of pro-regenerating macrophages, essential elements of skin regeneration.

[0217] Test 6 - Effect of the active ingredient according to the invention on the vascular system and the migration of endothelial cells.

[0218] The objective of this study is to evaluate the effect of the active ingredient according to the invention on the migration of aged human endothelial cells.

[0219] The endothelial cells that make up cutaneous microvessels are normally quiescent in capillaries. However, in response to a microenvironmental stimulus (hypoxia, injury), the endothelial cells are activated, migrate, and are able to remodel the vascular network to meet the needs of the damaged tissue. With age, senescent endothelial cells become dysfunctional, leading to defects in vascular remodeling and a decrease in the number of capillaries in the skin.

[0220] This study was performed using a time-lapse cell migration assay on young and old endothelial cells. The effect is measured by assessing the cells' ability to recolonize an area of ​​injury.

[0221] Young and old endothelial cells (HUVEC: Human umbilical vein endothelial cells) are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0222] The wound area is created. The cells are treated with secretomes from young fibroblasts (SFJ) or old fibroblasts (SFA) in the presence or absence of the active ingredient according to the invention at 0.25% and 1.00% (V / V). The cells are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0223] Visualization of endothelial cell migration is performed in "time-lapse" using a fully automated system which allows tracking of cell migration during culture.

[0224] The cell migration velocity within the wound is calculated using an internally developed image analysis script. The results are expressed in pm2 / min.

[0225] The results are given in Table 11.

[0226] [Tables 11] Migration velocity (pm2 / min) Ability to restore migration velocity (%) Young endothelial cells SFJ Control 1009 SFJ INVENTION 1.00% 1021 aged endothelial cells SFA Witness 532 -47 SFA INVENTION 0.25% 759 +48 SFA INVENTION 1.00% 975 +93

[0227] Migration of aged human endothelial cells is significantly reduced by 47% compared to young endothelial cells.

[0228] Tested on aged human endothelial cells, the secretome from aged human fibroblasts pretreated with 1.0% of the active ingredient according to the invention significantly restores the migration speed of endothelial cells by 93%.

[0229] The active principle according to the invention thus accelerates the migration of aged endothelial cells, essential for the regeneration of the (injured) vascular network.

[0230] Test 7 - Effect of the active ingredient according to the invention on the vascular system and the cohesion of endothelial cells.

[0231] The objective of this study is to evaluate the effect of the active ingredient according to the invention, within the fibroblast regenerating complex, on the synthesis of ZO-1 (Zonula Occludens -1), a junction protein between endothelial cells.

[0232] Vascular integrity is ensured by the establishment of adherens and tight junctions between endothelial cells. These junctions allow selective permeability of the vessels. ZO-1 is a constitutive component of endothelial tight junctions. With age, endothelial cells become dysfunctional and exhibit decreased expression and membrane targeting of tight junction components, leading to increased permeability.

[0233] The study was carried out by immunocytofluorescence, on young and old endothelial cells.

[0234] Young and old human umbilical vein endothelial cells (HUVEC) are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0235] The cells are treated with secretomes from young fibroblasts (SFJ) or old fibroblasts (SFA) in the presence or absence of the active ingredient according to the invention at 0.25% and 1.00% (V / V). They are then incubated in an atmosphere containing 5% CO2 at 37°C.

[0236] Immunocytological labeling of ZO-1 is carried out by fixation and permeabilization using primary antibody and secondary antibody coupled to a fluorophore.

[0237] Visualization is performed using a microscope coupled to an image analysis system.

[0238] The fluorescence area, corresponding to ZO-1 labeling, is proportional to the establishment of tight junctions between endothelial cells. An analysis Quantitative image analysis was performed using an internally developed image analysis script written in Python. The results are expressed in arbitrary units (AU).

[0239] The results are given in Table 12.

[0240] [Tables 12] ZO-1 Synthesis (UA) Capacity to restore ZO-1 synthesis (%) Young endothelial cells SFJ Control 6742 SFJ INVENTION 1.00% 7115 Old endothelial cells SFA Control 3232 -52 SFA INVENTION 0.25% 5003 +50 SFA INVENTION 1.00% 939 +77

[0241] The synthesis of ZO-1, a protein constituting intercellular junctions, is significantly reduced by 52% in aged endothelial cells compared to young cells.

[0242] Tested at 1.0% on aged endothelial cells, the active ingredient according to the invention significantly restores ZO-1 synthesis by 77%.

[0243] The active principle according to the invention thus makes it possible to strengthen cell cohesion.

[0244] Test 8 - Effect of the active ingredient according to the invention on the vascular system and the formation of the vascular network.

[0245] The objective of this study is to evaluate the effect of the active ingredient according to the invention on the formation of a capillary network of aged endothelial cells.

[0246] The remodeling of the cutaneous vascular network in adults in response to a stimulus (hypoxia, injury) occurs via angiogenesis. This process is defined as the formation of new vessels from pre-existing vessels. It results from the specification, migration, and proliferation of endothelial cells. Physiological angiogenesis allows the skin to heal properly. With age, capillary formation by angiogenesis is impaired, preventing proper healing and tissue homeostasis.

[0247] The study was carried out by immunocytofluorescence, on young and old endothelial cells using the specific marker of endothelial cells: PECAM-1 (CD31).

[0248] Young or old fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2.

[0249] The cells are treated with the 1.00% (V / V) extracts and then incubated at 37°C in an atmosphere containing 5% CO2.

[0250] Young and old human umbilical vein endothelial cells (HUVEC: Human Umbilical Vein Endothelial Cells) are seeded onto mats of young or old fibroblasts and incubated at 37°C in an atmosphere containing 5% CO2.

[0251] The cells are then treated with a 1.00% (V / V) extract and incubated at 37°C in an atmosphere containing 5% CO2.

[0252] Immunocytological labeling of CD31 is carried out by fixation and permeabilization using primary antibody and secondary antibody coupled to a fluorophore.

[0253] Visualization is performed using a microscope coupled with a camera and an image analysis system.

[0254] The fluorescence area corresponding to CD31 labeling is proportional to the extent of the vascular network formed by the endothelial cells.

[0255] A quantitative analysis of the images was carried out using an internally developed image analysis script. The results are expressed in arbitrary units (AU).

[0256] The results are given in Table 13.

[0257] [Tables 13] Ability to restore capillary network formation (%) Young endothelial cells Control INVENTION 1.00% +57 EXAMPLE 3 1.00% +28 Old endothelial cells Control INVENTION 1.00% +101 EXAMPLE 3 1.00% +77

[0258] Tested at 1.0% on aged endothelial cells, the active ingredient according to the invention significantly restores capillary network formation by 101%. Whereas Example 3 outside the invention shows lower efficacy.

[0259] The active ingredient according to the invention thus promotes angiogenesis.

[0260] Test 9 - _ Study of the biological activity of the active ingredient according to the invention in vivo.

[0261] The objective of this study is to evaluate in vivo the biological activity of the active ingredient according to the invention formulated at 2.5% in emulsion, compared to a placebo formula, on Caucasian and Asian volunteers.

[0262] The Caucasian panel consists of 19 healthy female volunteers, aged 70 to 78 years, with crow's feet wrinkles and a dull complexion.

[0263] The Asian panel consisted of 33 people aged 60 to 69 years and the placebo group of 32 people aged 60 to 68 years.

[0264] Effect of the active ingredient according to the invention on cell renewal.

[0265] Initially, the effect of the active ingredient according to the invention was evaluated at the level of the forearms on the Caucasian panel by studying its ability to accelerate cell renewal after 14 days of twice-daily application.

[0266] Cell renewal is measured by the skin's ability to eliminate DHA (dihydroxyacetone), which is a protein-staining marker. Cell renewal results in the disappearance of the DHA-induced staining.

[0267] A gel containing 8% DHA is applied to the skin surface of volunteers. Monitoring of the color obtained after application of the DHA gel begins 72 hours after staining on untreated areas, areas treated twice daily with a placebo formula, or areas treated twice daily with a formula containing 5% of the active ingredient according to the invention. The evaluation is performed using photographs of the skin taken at regular intervals over 14 days with a dermatoscope.

[0268] The results corresponding to the effect of the active ingredient according to the invention formulated at 2.5% in emulsion, on the ability to eliminate DHA is presented in [Fig.1].

[0269] Compared to placebo, the active ingredient according to the invention, formulated at 2.5% in emulsion, promotes epidermal dynamics by increasing the skin's ability to eliminate DHA.

[0270] This effect is observed and significant from 2 days after application of the products and continues throughout the study (D14).

[0271] Indeed, with the active ingredient according to the invention, in less than 7 days, the DHA staining is eliminated by more than 80%, compared to 11 days for the area treated with the placebo.

[0272] Effect of the active ingredient according to the invention on the structure of the dermis.

[0273] Subsequently, the effect of the active ingredient according to the invention on the structure of the dermis was evaluated on the face after 21 and 42 days of application by assessing the quality of the matrix using LC-OCT (Line-Field Optical Coherence Tomography). This study was conducted on Caucasian and Asian panels.

[0274] The results showing the effect on the Caucasian panel of the active ingredient according to the invention on the quality of the dermal matrix evaluated over a thickness of 50 pm under the JDE are presented in Table 14 below.

[0275] [Tables 14] D21 D42 Placebo -2 -1 INVENTION 2.5% 30 27

[0276] From 21 days of treatment, the active ingredient according to the invention formulated at 2.5% significantly improves the quality of the matrix of Caucasian volunteers by 32% compared to placebo.

[0277] This effect was observed in 89% of subjects. Thus, the matrix dynamics of the dermis are restored. This effect persists after 42 days of treatment.

[0278] The results showing the effect of the active ingredient according to the invention on the quality of dermal fibers of Asian volunteers are presented in Table 15 below.

[0279] [Tables 15] D21 D42 Placebo 1 0 INVENTION 2.5% 21 18

[0280] After 21 days of treatment, the active ingredient according to the invention, formulated at 2.5%, significantly improves the fiber quality of Asian volunteers by 20% compared to placebo. This effect was observed in 97% of the Asian subjects. This effect persists after 42 days of treatment.

[0281] Trial 10 - Evaluation of the cosmetic benefits of the active ingredient according to the invention

[0282] The objective of this study is to evaluate in vivo the cosmetic benefits of the active ingredient according to the invention formulated at 2.5% in emulsion, compared to a placebo formula, in Caucasian and Asian volunteers. The effect of the active ingredient according to the invention was evaluated after 21 and 42 days of twice-daily application.

[0283] The Caucasian panel consists of 19 healthy female volunteers, aged 70 to 78 years, with crow's feet wrinkles and a dull complexion.

[0284] The Asian panel consists of 33 people aged 60 to 69 and the group placebo of 32 people aged 60 to 68 years.

[0285] Action on the biomechanical properties of the skin on a Caucasian panel

[0286] The results corresponding to the effect of the active ingredient according to the invention on the The biomechanical properties of the skin of Caucasian volunteers are presented in [Fig.2],

[0287] The active ingredient according to the invention, compared to placebo, improves the biomechanical properties of the skin of Caucasian volunteers.

[0288] Indeed, after just 21 days of treatment, the active ingredient according to the invention increases skin firmness by: - ​​10%, an effect observed in 84% of subjects - 14% of skin elasticity, an effect observed in 68% of subjects - 25% of skin tone, effect observed in 84% of subjects.

[0289] Furthermore, 79% of volunteers found their skin firmer and 79% more toned with the active ingredient according to the invention, compared to 68% and 74% respectively for the placebo group. This effect persisted after 42 days of twice-daily application.

[0290] Anti-wrinkle action on a Caucasian panel

[0291] The results corresponding to the anti-wrinkle effect of the active ingredient according to the invention evaluated on the crow's feet area of ​​Caucasian volunteers by visual scoring on digital photographs compared to placebo are presented in Table 16.

[0292] [Tables 16] D21 D42 Placebo 0 2 INVENTION 2.5% -10 -8

[0293] After 21 days of treatment, compared to placebo, the active ingredient according to the invention, formulated at 2.5% in emulsion, exhibits an anti-wrinkle effect, significantly reducing the stage of crow's feet wrinkles by 10%. This effect is observed in 74% of volunteers.

[0294] Furthermore, 74% of subjects found that their wrinkles and fine lines were less visible with the active ingredient according to the invention, compared to only 58% for the placebo treatment. This effect persisted after 42 days of treatment.

[0295] Action on the cutaneous microvascular network of the Caucasian panel

[0296] The results corresponding to the effect of the active ingredient according to the invention on the visibility of micro-vessels in the skin of Caucasian volunteers are presented in Table 17.

[0297] [Tables 17] D21 D42 Placebo -5 -14 INVENTION 2.5% -10 -21

[0298] From 21 days of treatment and compared to placebo, the active ingredient according to the invention formulated at 2.5% reduces the visibility of dystrophic vessels in the cheeks (-6%).

[0299] This effect continues after 42 days of treatment where a significant decrease of 7% is observed; effect validated in 63% of subjects.

[0300] By reducing the visibility of dystrophic microvessels, the active principle according to the invention improves cutaneous vascularization.

[0301] Action on dark circles in the Caucasian panel

[0302] The results corresponding to the effect of the active ingredient according to the invention on the dark circles of Caucasian volunteers are presented in Table 18 below.

[0303] [Tables 18] D21 D42 Placebo -0.3 1.6 INVENTION 2.5% -3.1 -1.6

[0304] Compared to placebo, the active ingredient according to the invention, formulated at 2.5% in emulsion, reduces dark circles by decreasing parameter b* by 2.9% after 21 days of application and this effect continues until the end of the study. Parameter b* represents the range from blue to yellow in colorimetric measurements; it is characteristic of yellow melanin pigmentation in the skin. The lower this parameter, the lighter the skin appears.

[0305] This effect is visible in 68% of the volunteers and 79% of them perceived the product as a concealer compared to only 53% for the placebo treatment.

[0306] Action on the radiance of the complexion of the Caucasian panel

[0307] The results corresponding to the effect of the active ingredient according to the invention formulated at 2.5% on the main characteristic parameters of the radiance of the complexion evaluated by experts on the Caucasian panel, are presented in Table 19 below.

[0308] [Tables 19] J21 J42 Radiation 9 10 Pink color 15 16 Olive color -7 -11 Eye fatigue -6 -8

[0309] After 21 days of twice-daily application and compared to placebo, the active ingredient according to the invention, formulated at 2.5% in emulsion: • gives a brighter and fresher complexion by significantly increasing skin radiance by 9% and pink color by 15%. Effect observed in 74% and 84% of volunteers respectively. • Significantly reduces olive color by 7% and eye fatigue by 6%. This effect was observed in 79% and 89% of volunteers, respectively

[0310] . This effect is supported by the volunteers' evaluation: 84% at day 21 found their skin more radiant with the active ingredient according to the invention versus 63% for the placebo treatment.

[0311] Anti-wrinkle action on an Asian panel

[0312] The results corresponding to the anti-wrinkle effect of the active ingredient according to the invention studied by visual scoring by experts on Asian volunteers are presented in Table 20. A jury of 3 people evaluated the stage of crow's feet wrinkles in each photo on a pre-defined scoring scale.

[0313] [Tables20] D21 D42 Placebo 1 -1 INVENTION 2.5% -5 -7

[0314] After 21 days of treatment, compared to placebo, the active ingredient according to the invention, formulated at 2.5% in emulsion, exhibited an anti-wrinkle effect in Asian volunteers, significantly reducing the stage of crow's feet wrinkles by 6%. This effect was observed in 58% of the volunteers. This effect persisted after 42 days of treatment.

[0315] Conclusion

[0316] The active ingredient according to the invention, formulated at 2.5% in emulsion, provides cosmetic benefits from 21 days of treatment which continue after 42 days of treatment.

[0317] Indeed, the active ingredient according to the invention allows: - an improvement in the biomechanical properties of the skin - a reduction in wrinkles in Caucasian and Asian volunteers - an improvement in skin vascularization, by reducing dystrophic microvessels and dark circles - an improvement in the radiance of the complexion.

Claims

Claims

1. Cosmetic active ingredient comprising at least one extract of Saccharomyces cerevisiae, characterized in that said extract corresponds to the cytosolic fraction comprising oligoglucans with a molar mass of between 2 and 5 kDa.

2. Cosmetic active ingredient according to the preceding claim, characterized in that the oligoglucans represent between 10% and 35% by weight of dry matter of the extract.

3. Cosmetic active ingredient according to the preceding claim, characterized in that the extract comprises a glucose / mannose ratio greater than 60%.

4. Cosmetic active ingredient according to one of the preceding claims, characterized in that the extract is a mechanical lysate of Saccharomyces cerevisiae.

5. Cosmetic active ingredient according to one of the preceding claims, characterized in that said extract is capable of being obtained by a process comprising the following steps: a. solubilization of at least 50 g / L of Saccharomyces cerevisiae in water by mechanical homogenization, b. separation of the soluble and insoluble phases c. ultrafiltration and recovery of the soluble filtrate d. molecular sorting by ion chromatography.

6. Cosmetic composition for topical application comprising at least one active ingredient according to one of the preceding claims and a physiologically acceptable medium.

7. Cosmetic composition according to the preceding claim, in which the active ingredient in liquid form represents at least 0.1% by weight of the total weight of the composition.

8. Cosmetic use of an active ingredient according to one of claims 1 to 5 or of a cosmetic composition according to one of claims 6 or 7, to improve skin regeneration linked to normal skin aging.

9. Cosmetic use according to the preceding claim, characterized in that the extract or composition improves the interconnectivity of the cutaneous, vascular and immune systems.

10. Cosmetic use according to the preceding claim, characterized in that the extract or composition increases the synthesis and secretion of KGF, EGF, IGF, FGF, PDGF, TGF-[3 and VEGF.

11. Cosmetic use according to one of claims 8 to 10 for improving the radiance of the complexion.

12. Cosmetic use according to one of claims 8 to 11 for preventing and / or reducing dark circles.

13. Cosmetic use according to one of claims 8 to 12 for preventing and / or reducing wrinkles.

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