ECO-BIOLOGICAL COMPOSITION COMPRISING A COMBINATION OF ACTIVE INGREDIENTS CAPABLE OF PROTECTING SKIN COLLAGENS

Abstract

The present invention relates to a cosmetic composition, advantageously eco-biological, comprising an apple seed extract, at least one phytosterol, and at least one exopolysaccharide. In particular, the composition of the invention comprises a mixture of an apple seed extract and at least one phytosterol, said mixture being combined with at least one exopolysaccharide; as well as the use of this composition to reduce the loss of collagen functionality, to combat skin aging, or to regenerate the skin. [Fig. 1]

Description

Title of the invention: ECO-BIOLOGICAL COMPOSITION COMPRISING A COMBINATION OF ACTIVE INGREDIENTS CAPABLE OF PROTECTING SKIN COLLAGENS Scope of the invention

[0001] The present invention relates to the field of cosmetics, in particular to cosmetic compositions, advantageously ecobiological, and their uses to reduce, or even prevent / stop, the proteolysis of proteins constituting the skin such as collagens, to fight against skin aging or to ensure or promote skin healing. State of the art

[0002] The skin consists of three superimposed layers: the epidermis, the dermis, and the hypodermis. The epidermis is a stratified epithelium composed primarily of layers of keratinocytes, which represent 90 to 95% of epidermal cells. The epidermis provides a barrier function, combating various physical, chemical, and microbial aggressions from the surrounding environment while limiting transepidermal water loss (TEWL). The dermis is mainly composed of fibroblasts dispersed in the extracellular matrix (ECM). This ECM is composed of a set of polysaccharides (i.e., glycosaminoglycans, generally covalently linked to a protein to form proteoglycans) and proteins. The protein component of this ECM is characterized by the presence of a first group, consisting of collagens and elastin, and a second, less abundant group, consisting of fibronectin and laminin.Essentially, the first group of proteins is responsible for the structure of the ECM, while the second group of proteins is involved in the organization of the ECM structure and in cell-matrix adhesion.

[0003] Among the proteins of the ECM, collagens represent more than 75% of the dry weight of human skin and their function is to give tissues mechanical resistance to stretching or traction (tension). To date, at least 28 different types of collagen have been identified, distinguished by their basic morphology, amino acid composition, and physical properties. Each type of collagen has its own structure and is found in specific organs. For example, type I collagen (also called collagen I, COL1A1, or COL1A2) represents 90% of the collagen in a vertebrate and constitutes the framework of bone and connective tissues (e.g., skin, tendons, cornea, or internal organs). In parallel, type XVII collagen (also called collagen XVII, COL17Al, or BPAG2 for bullous) pemphigoid antigen (cGIA) is a 180 kDa transmembrane protein, a component of hemidesmosomes, and a component that anchors keratinocytes to the dermo-epidermal junction (Nishizawa et al., 1993). Therefore, a reduction in physiological levels of collagen XVII has negative effects on epidermal structure. More specifically, it has been associated with a loss of hemidesmosomes' stability and, consequently, epidermal thinning, which compromises adhesion between the epidermis and the dermis (Mondon et al., 2015; Langton et al., 2016). Furthermore, overexpression of human collagen XVII in vivo in aged mice has been shown to prevent age-related epidermal thinning (Liu et al., 2019).

[0004] Collagen XVII is also involved in cutaneous pigment homeostasis and in the maintenance of hair follicle stem cells and melanocyte stem cells located in the hair follicle (Shirai et al., 2019). The importance of collagen XVII in maintaining cutaneous homeostasis has also been demonstrated by gene knockout of the COL17A1 gene in a mouse model, which then exhibits pigmentation abnormalities similar to those observed during aging in humans. Furthermore, repeated exposure to UVB also induces hypopigmentation similar to that observed in normal mice with collagen XVII deficiency. These pigmentary abnormalities do not appear when human collagen XVII is overexpressed in a mouse model.Furthermore, it has also been shown that the healing of deep wounds in aged mice is comparable to that of young mice in which the expression of the COL17A gene is inhibited. These results demonstrate that hemidesmosomes' instability, due in particular to the loss of collagen XVII, leads to a delay in wound healing during aging (Liu et al., 2019).

[0005] In humans, several studies have shown that cutaneous levels of collagen XVII decrease with age (Langton et al., 2016; Mondon et al., 2015; Watanabe et al., 2017). Generally, however, this decrease does not appear to be related to a reduction in gene expression (Watanabe et al., 2017; Langton et al., 2016). The progressive reduction in collagen XVII levels in the skin therefore appears to be due to a loss of the protein, or at least of the functional protein, with age.

[0006] It is clear from these elements that collagen XVII is an important link in cutaneous homeostasis and that the reduction of its content in the skin, which normally occurs during aging, leads to epidermal disturbances, including destabilization of hemidesmosomes, pigmentary and hair abnormalities as well as delays in healing.

[0007] There therefore remains a clear need to identify a reliable and cosmetic, advantageously eco-biological, easy-to-use, inexpensive, and easily integrated into daily routine means of reducing, slowing down, or even preventing or stopping the drop in skin collagen levels, particularly collagens I and XVII, to ultimately fight against skin aging and / or promote skin healing.

[0008] Description of the invention

[0009] After much research, it is to the applicant's credit that she was able to identify, unexpectedly and surprisingly, a composition comprising an association of compounds / active ingredients that act in synergy to meet this need.

[0010] In accordance with the ecobiological approach to acting on the biological causes of skin disorders, an objective of the present invention is to combat the decrease in collagen levels in the ECM, in particular collagen I and XVII, which ultimately helps to combat skin aging, the loss of stability of hemidesmosomes, thinning of the epidermis, the appearance of wrinkles and fine lines, the appearance of pigmentary and / or hair abnormalities, and to ensure the maintenance of skin homeostasis.

[0011] Thus, a first object of the present invention relates to a cosmetic composition, advantageously ecobiological, comprising an apple seed extract, at least one phytosterol and at least one exopolysaccharide, advantageously of bacterial origin.

[0012] In particular, the invention relates to a cosmetic composition, advantageously ecobiological, comprising a mixture of an apple seed extract and at least one phytosterol, said mixture being combined with at least one exopolysaccharide, preferably of bacterial origin.

[0013] The present invention offers various advantages, including the fact that it allows the simple and routinely integrable use of an effective composition whose compounds act in synergy on the aforementioned causes of skin dysfunctions, which is respectful of the communities of living cells constituting the skin and which constantly interact with each other and with their environment to maintain the balance of this skin ecosystem, and which is inexpensive.

[0014] In particular, the present invention makes it possible to combat the loss of collagen functionality, specifically: - to combat the proteolysis of collagens, particularly collagens I and XVII, advantageously collagen XVII; and / or - to combat the glycation of collagens, in particular collagens I and XVII, advantageously collagen I.

[0015] In the context of the invention, the expression "ecobiological composition, active ingredient or excipient" means compositions, ingredients or excipients that are respectful of people, their interactions with the world, and the planet. In particular, it refers to a composition, an active ingredient or an excipient res protective of the communities of living cells that make up the skin (i.e., skin microbiota, keratinocytes, fibroblasts, etc.) and which constantly interact with each other and with their environment to maintain the homeostasis / balance of this skin ecosystem.

[0016] In the context of the invention, the term "ecobiological approach" refers to the particular approach initiated and developed by the inventor which combines skin biology and ecology to help the skin live according to its natural biology, in the long term.

[0017] In the context of the invention, the terms "phytosterols," "plant sterols," and "natural sterols" are used interchangeably and refer to a group of sterols naturally produced by plants that have a chemical structure similar to cholesterol. These phytosterols are components of plant cell membranes and include, in particular, the following compounds: campesterol, beta-sitosterol, stigmasterol, brassicasterol, campestanol, and sitostanol.

[0018] In the context of this invention, the term "exopolysaccharide" or "EPS" refers in particular to polysaccharides excreted by microorganisms into the external environment. EPS are high molecular weight molecules that exhibit a wide diversity of chemical structures due to the different sugars that can compose the polymers, the possible presence of substituents such as sulfate, acetate, pyruvate, and succinate groups, as well as the polymer structure itself, which can be branched or linear. This high variability gives them particular properties that are of interest in various fields such as the food industry and the chemical industry.

[0019] In the context of the invention, the terms "prevent", "prevent", "avoid", "stop" or "inhibit" are used interchangeably with regard in particular to "the loss of collagen functionality" and refer to taking measures aimed at reducing or totally eliminating the probability of occurrence of said loss of protein functionality and / or measures aimed at limiting the extent and / or severity of the consequences of a loss of collagen functionality.

[0020] In the context of the invention, the expressions "loss of protein functionality" and "inactive protein" are used interchangeably and refer to a loss of biological activity of a protein originating, for example, from a spatial disorganization / loss of folding induced by denaturing agents of a physical nature (i.e. modification of temperature, pH or irradiation) or chemical nature (i.e. oxidizing agents, thiol reducing agents, detergents or heavy metal ions).

[0021] In the context of the invention, "collages" refers to a protein composed of three alpha polypeptide chains, themselves mainly composed of three types of amino acids (i.e., glycine, proline, and hydroxyproline). These three chains are linked through hydrogen bonds and covalent bonds. Each type of collagen (type I to XXVIII collagen) has its own structure and is found in specific organs or tissues.

[0022] In the context of the invention, "healing" or "to heal" refers to the various phases of the process by which tissue damage is repaired / the integrity of damaged tissue is restored following an injury. The different stages of this process are: - an inflammatory phase: phagocytosis and elimination of bacteria and other dirt, and the factors that produce the migration and division of cells that participate in the proliferative phase are released; - a proliferative phase: angiogenesis, increased collagen content, formation of granular tissue, epithelialization and wound contraction; and - a maturation and remodeling phase: collagen is remodeled and realigned along the lines of tension and cells that are no longer needed are eliminated by apoptosis.

[0023] In the context of the invention, the term "active ingredient" refers to a substance or compound possessing biological and / or therapeutic properties that underlie a physiological effect. The active ingredient is to be distinguished from at least one excipient present in the composition according to the invention.

[0024] In the context of the invention, "molecular weight (Mw)" refers to the average molecular weight by weight (Mw or Molecular Weight) of a compound, of a protein.

[0025] In the context of the invention, "composition for cutaneous application" or "for topical use" means a composition compatible with application to the skin, mucous membranes, hair and / or scalp, preferably human skin.

[0026] Preferably, the present invention relates to a cosmetic composition, advantageously eco-biological, comprising an apple seed extract, at least one phytosterol, and at least one exopolysaccharide, preferably of bacterial origin. Advantageously, the composition according to the invention has the following additional technical characteristics, taken alone or in combination: - Apple seed extract contains quercitrin, oleanolic acid and / or ursolic acid; - Apple seed extract represents between 0.0000001% to 5% by total weight of the composition, advantageously between 0.000001% and 0.3%; - phytosterol is derived from rapeseed extract; - phytosterol is selected from the group consisting of campesterol, beta-sitosterol, stigmasterol, brassicasterol, campestanol and sitostanol; - Phytosterols represent between 0.0000001% and 5% by weight of the total composition, advantageously between 0.000001% and 0.1%; - the exopolysaccharide is advantageously of bacterial origin, preferably from the marine bacterium Vibrio alginolyticus; - the exopolysaccharide represents between 0.0000001% to 5% by total weight of the composition, advantageously between 0.000001% and 0.1%; - the exopolysaccharide has a molecular weight (Mw) greater than 300 kDa, advantageously greater than 1 MDa;

[0027] -the exopolysaccharide is composed by the repetition of a tetrasaccharide consisting of three units of galacturonic acid and one unit of N-acetylglucosamine in which two of the three units of galacturonic acid are substituted by an alanine group and a serine group; - the ratio between the weight of the apple seed extract and the weight of at least one phytosterol is between 0.1 and 2, advantageously between 0.3 and 1.5, preferably between 0.7 and 0.9, more advantageously 0.8; - the ratio between the weight of the mixture of apple seed extract and phytosterol and the weight of the exopolysaccharide is between 0.1 and 2, advantageously between 0.3 and 1.5, preferably between 0.5 and 0.6; - Apple seed extract corresponds to the INCI designation PYRUS MALUS (APPLE) SEED EXTRACT; - Apple seed extract is lipophilic or hydrophilic, advantageously lipophilic; - Apple seed extract includes at least one flavonoid and / or isoflavonoid and / or one of their derivatives; - phytosterol corresponds to the INCI designation BRASSICA CAMPESTRIS (RAPESEED) STEROLS; - the exopolysaccharide corresponds to the INCI designation SACCHARIDE ISOMERATE; - the composition also includes vitamin PP, corresponding to the INCI designation Niacinamide, or one of its derivatives; - vitamin PP represents between 0.0001% and 10% by total weight of the composition, advantageously between 0.001% and 5%, preferably between 0.1% and 2%; - Apple seed extract, phytosterol, exopolysaccharide and vitamin PP, when included in the composition, are the sole active ingredients in the composition; the composition also includes a compound corresponding to the INCI designation HEXYLDECANOL; - the compound corresponding to the INCI designation HEXYLDECANOL represents between 0.0000001% and 5% by total weight of the composition, advantageously between 0.000001% and 0.3%; - the composition also includes a compound corresponding to the INCI designation tocopherol; - the compound corresponding to the designation tocopherol represents between 0.0000000002% to 0.01% by total weight of the composition, advantageously between 0.000000002% and 0.0006%; - compounds with the INCI designations hexyldecanol and TOCOPHEROL, when included in the composition, are solely excipients; - the combination of apple seed extract, at least one phytosterol and compounds corresponding to the INCI designations hexyldecanol and TOCOPHEROL are combined in a single raw material; - the combination of compounds corresponding to the INCI designations PYRUS MALUS (APPLE) SEED EXTRACT, BRASSICA CAMPESTRIS (RAPESEED) STEROLS, hexyldecanol and TOCOPHEROL are combined in a single raw material; - the EDERLINE™ S cosmetic raw material, corresponding to the INCI designation Hexyldecanol (and) butylene glycol (and) Pyrus malus (apple) seed extract, or the EDERLINE™ LS raw material, corresponding to the INCI designation hexyldecanol (and) Pyrus malus (apple) seed extract (and) Brassica campestris (rapeseed) sterols (and) tocopherol, marketed by the company ASHLAND, can be used as a source of apple seed extract; - the raw material GENEROL™R corresponding to the INCI designation Brassica campestris (rapeseed) sterols marketed by BASF Corporation, or the raw material EDERLINE™ LS, corresponding to the INCI designation hexyldecanol & Pyrus malus (apple) seed extract & Brassica campestris (rapeseed) sterols & tocopherol marketed by ASHLAND, can be used as a source of phytosterol; - the raw materials EPIDERMIST POWDER 4 0 PUR, EPS SEAMAT POWDER PUR, EPS SEAWHITE PA corresponding respectively to the INCI designations saccharide isomerate, saccharide isomerate and Water (and) Saccharide isomerate (and) Phenethyl alcohol, marketed by the company CODIF, can be used as a source of exopolysaccharide; - the raw material NIACINAMIDE corresponds to the INCI designation NIA-CINAMIDE marketed by the company QUIMICA, can be used as a source of vitamin PP; - the composition further comprises a bioactive system combining, on the one hand, a stable form in aqueous solution of a nucleotide chosen from ATP (adenosine triphosphate), Gp4G (diguanosine tetraphosphate) and Ap4A (diadenosine tetra- phosphate); and on the other hand, at least one biomimetic peptide comprising at most six amino acids, mimicking a skin polypeptide or a skin protein, or a biomolecule that is an agonist or antagonist of said peptide or protein. This bioactive system catalyzes the metabolic activity of skin cells while achieving a dermocosmetic or therapeutic effect through the use of biomimetic peptides. These peptides can be selected to obtain the desired effect, for example, an inhibition of irritations of neurogenic origin, a depigmenting effect, an effect inhibiting any intolerance or sensitization, etc.; - in the bioactive system, the nucleotide represents at most 10% by total weight of the composition, preferably between 0.001% and 5%; and the biomimetic peptide represents between 0.001% and 1% by total weight of the composition; - the composition further includes compounds that can contribute to internal protection through an action that may consist of DNA protection, a reduction of immunosuppression induced by UV radiation, an antiradical action or a combined effect of these actions; - the composition further comprises glycyrrhetinic acid, a derivative or a salt of this acid, used as a soothing agent (anti-inflammatory agent) and representing between 0.01% and 2% by total weight of the composition, advantageously between 0.1% and 1%; - the composition further comprises a cellular structure-preserving free radical scavenging agent, such as, for example, vitamin E and / or its fat-soluble or water-soluble derivatives, in particular tocotrienol and / or tocopherol, representing between 0.001% and 10% by total weight of the composition, advantageously between 0.02% and 2%, preferably 0.04%; - the composition further comprises a p53 protein protective agent, such as epigallocatechin gallate (EGCG), representing between 0.001% and 0.1% by total weight of the composition, advantageously between 0.005% and 0.05%; - the composition further comprises at least one compound selected from the group consisting of: fructooligosaccharides, boldo extracts, hyaluronic acid, hyaluronic acid derivatives, carnosine and adenosine; - adenosine represents between 0.0001% and 5% by total weight of the composition, advantageously between 0.001% and 1%, preferably between 0.01% and 0.05%; - Adenosine is in a non-phosphated form, advantageously in powder form, and corresponds to the INCI designation Adenosine marketed by the company PHARMA WALDHOF; - carnosine represents between 0.001% and 5% by total weight of the composition, advantageously between 0.01% and 1%; - Carnosine is in purified or highly purified form, advantageously obtained by chemical synthesis, and can be the raw material for Kopsin corresponding to the de INCI name Carnosine marketed by KUMAR ORGANIC PRODUCTS LTD; - Fructo-oligosaccharides, also called oligofructoses or FOS, represent between 0.001% and 10% by total weight of the composition, advantageously between 0.01% and 5%; - FOS are in purified or highly purified form, and correspond to the fructooligosaccharides raw material corresponding to the INCI name FRUCTOOLL GOSACCHARIDES marketed by the company Shaanxi Bolin Biotechnology Co; - Hyaluronic acid has a molecular weight (Mw) greater than 300 kDa, advantageously greater than 1 MDa and is present in acid form, as a salt, in particular sodium, potassium or magnesium salts, preferably sodium salts; - Hyaluronic acid represents between 0.0001% and 10% by total weight of the composition, advantageously between 0.001% and 2%, preferably between 0.01% and 1%; - hyaluronic acid corresponds to the raw material BIOSODIUM HYA-LURONATE MMW corresponding to the INCI name Sodium hyaluronate marketed by the company DKSH; - Boldo (Peumus boldus) or boldo extract is a species of tree in the Monimiaceae family and whose leaf extracts induce the synthesis of defensins and cathelicidins, boldo extract represents between 0.0001% and 10% by total weight of the composition, advantageously between 0.001% and 2%, preferably between 0.01% and 1%; - boldo extract corresponds to the raw material MP BETAPUR PGF corresponding to the INCI name Water (and) Butylene Glycol (and) Peumus Boldus Leaf Extract (and) Pentylene Glycol (and) Xanthan Gum marketed by the company BASF; - the composition also includes peptide extracts of soy and / or wheat; - peptide extracts of soy and / or wheat represent between 0.01% and 20% by total weight of the composition, advantageously between 0.1% and 10%, preferably between 0.2% and 0.7%; - the composition further comprises peptide extracts of soy and wheat in a weight ratio respectively between 80 / 20 and 20 / 80, advantageously between 70 / 30 and 30 / 70, preferably equal to 60 / 40; - the peptide extracts of soy and / or wheat are free of synthetic tripeptides GHK (glycyl-histidyl-lysine; INCI: Tripeptide-1); - the soy peptide extract is the extract identified under CAS number 68607-88-5 and the wheat peptide extract is the extract identified under CAS number 70084-87-6, advantageously the wheat and soy extracts correspond to the INCI designations Hydrolyzed wheat protein and Hydrolyzed soy protein, respectively; - the composition is in a galenic form suitable for acceptable cosmetic use, i.e. compatible with the skin, mucous membranes, hair and scalp; - the composition is in a pharmaceutical form suitable for cutaneous application and / or topical use; and / or - The composition is in the form of an aqueous, hydroalcoholic, organic, or oily solution; a suspension or dispersion in solvents or fatty substances, such as a lotion or serum; a vesicular dispersion; a water-in-oil (W / O), oil-in-water (O / W), or multiple emulsions such as a water-in-oil-in-water (W / O / W) emulsion. The emulsion may be more or less thick and may be in the form of a cream or lotion; the composition of the invention may also be in the form of an ointment, gel, solid stick, anhydrous paste or solid products, foam, including aerosol, a biphasic composition, or a sprayable composition.

[0028] The pharmaceutical form of the composition and its method of preparation, and consequently the excipients suitable for the composition of the invention, can be chosen by a person skilled in the art based on their general knowledge, depending on the type of composition desired. In particular, the composition may include any fat commonly used in the cosmetic field. Examples include silicone-based fats such as silicone oils, gums, and waxes, as well as non-silicon-based fats such as oils and waxes of vegetable, mineral, animal, and / or synthetic origin. The oils may be volatile or non-volatile. Other examples include hydrocarbons, synthetic esters and ethers, fatty alcohols, and fatty acids.The composition may also include an aqueous medium, a hydroalcoholic medium containing an alcohol such as ethanol or isopropanol, or an organic medium comprising common organic solvents such as Cl-6 alcohols, particularly ethanol and isopropanol, glycols such as propylene glycol, and ketones. Naturally, those skilled in the art will take care to choose any additional adjuvants or excipients, and / or their quantity, in such a way that the advantageous properties of the composition according to the invention are not, or not substantially, altered by the intended addition. The composition may include at least one conventional emulsifier, chosen from amphoteric, anionic, cationic, or nonionic emulsifiers, used alone or in mixtures. It may be particularly advantageous to formulate the composition of the invention so that it is sprayable.This can be achieved, for example, by formulating specific emulsions comprising particular combinations of excipients.

[0029] Another object of the present invention also relates to the cosmetic use of the composition as described above, to reduce, or even prevent / stop the loss of collagen functionality, advantageously the loss of functionality induced by proteolysis, glycation, acetylation, acylation of collagens, advantageously proteolysis or glycation, preferably collagen I and / or collagen XVII.

[0030] In particular, this cosmetic use of the composition of the invention makes it possible to: - to combat proteolysis, glycation, acetylation or acylation of collagen I; - combat glycation, proteolysis, acetylation, and acylation of collagen XVII; - combat glycation of collagen I; and / or - to combat the proteolysis of collagen XVII.

[0031] In particular, this cosmetic use of the composition of the invention makes it possible to: - combat the proteolysis of collagens, particularly collagens I and XVII, advantageously collagen XVII; and / or - to combat the glycation of collagens, in particular collagens I and XVII, advantageously collagen I.

[0032] Another object of the present invention also relates to the cosmetic use of the composition as described above, to combat skin aging, advantageously to combat the appearance of wrinkles.

[0033] Another object of the present invention also relates to the cosmetic use of the composition as described above, to regenerate the skin, advantageously to promote skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0034] Another object of the present invention also relates to the composition as described above for use in reducing, or even preventing, the loss of functionality of collagens, advantageously the loss of functionality induced by proteolysis, glycation, acetylation, acylation of collagens, advantageously proteolysis, preferably of collagen I and / or collagen XVII, even more advantageously of collagen XVII.

[0035] In particular, the composition of the invention for its use described above makes it possible to: - combat the proteolysis of collagens, particularly collagens I and XVII, advantageously collagen XVII; and / or - to combat the glycation of collagens, in particular collagens I and XVII, advantageously collagen I.

[0036] Another object of the present invention also relates to the composition as described above for use in combating skin aging, advantageously for combating the appearance of wrinkles.

[0037] Another object of the present invention also relates to the composition as described above for use in regenerating the skin, advantageously to promote skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0038] The manner in which the invention can be implemented and the resulting advantages will be more apparent from the following embodiment examples, given by way of illustration and not limitation, in support of the attached figure. Figures

[0039] [Fig-1]: Evaluation of the protection of Pyrus malus (Apple) Seed Extract, Brassica Campestris sterols and saccharide isomerate and their combinations against collagen-XVII proteolysis in a culture of normal human keratinocytes. The results are from a 3-well experiment per condition. Statistics: Student's t-test compared to the untreated condition or solvent control condition; # Student's t-test for saccharide isomerate compared to Pyrus malus (apple) seed extract + Brassica campestris sterols + saccharide isomerate; * or #: 0.05 < p-value < 0.01; ** or ##: 0.05 < p < 0.01; and *** or ###: p < 0.005.

[0040] EXAMPLES OF IMPLEMENTATION

[0041] The percentages indicated are given in weight of product relative to the total weight of the composition in the tables below.

[0042] Example 1: Cosmetic composition according to the invention - O / W emulsion

[0043] A composition according to the invention is described in Table 1.

[0044] [Tables 1] Desfgiîstiü» ff / a % Aqua / Water / Eau Qsp 180 Gjycerûs 30.00 CocoOpryiate 3.00 Propy Ihepiyi c ap ry i ate 3.3025 Tftbgtenic P£G-20 Esters 230 Cfamclrvs crispus extract 1.00 Fragrance {Parfum} 0.30 HyaLrontocid 0.30 Hexyidecanoi 0383 Dispdtym EDTA 030 wr (Soybean} Os! 0.1875 Acrylate$ / CW-80 Atkyt Acryiate Crosspolymer 0.15 Cil rie Acid 0.08 Sodium MsUbiWite 0.05 Niadnamide 0.0825 7« would cost 0.0101 GîyçineSqa (Soybeao) Sterois 0.01 as Frud ooiigosaccha ri des 0.0509 Saccharide isomerate 0.02 Pyras matas (apple] seed extract 0.006 Smsska campestris (rapesaed) sterois 0.0048

[0045] Example 2: Cosmetic composition within the meaning of the invention - GEL

[0046] A composition according to the invention is described in Table 2.

[0047] [Tables 2] Designation ÎNG ggggS|WiiSSSSSSS Aqua / Water / Eau Qsp WD Giycerin 2.00 Hydroxyethyi Acn / iate / Sodium 1.125 Acryioyldimethyi Taurate Copoiymer isohexadscane 0.765 Titanium Dioxide 0.375 Phenoxyethanoi 0.35 1,2-hexanedtoi 0.2 G Polysorbate 20 0.165 Sacoharice isomerate 0.00 Pyrus mains (appie) seed extract 0.025 Brassica campestris jrapesesd) sterois 0.020

[0048] Example 3: Cosmetic composition within the meaning of the invention -SprAY

[0049] A composition according to the invention is described in Table 3.

[0050] [Tables3] INCI designation % Aqüa / water / eau Qsp 100 Giycerîn 15.00 Dipropylene giycol 9.00 isostearyi isostearate 2.00 Propyiheptyl capryîate 1.9906 Squaiane 1.00 Sodium coçoyi glutamate 0.8825 Niadnamide 0.50 Pentyiene gîycoi 0.50 Hexyldecanoi 0.294 Disodium EDTA 0.20 Gtric acid 0.17 Propanediol 0.125 Xanthan gum 0.10 Tocophero! 0.036 Lecithin 0.0201 Helianthus annuus [sunflower) seed oil 0.015 Saccharide isomate 0.01 Pyrus malus (appie) seed extract 0.003 Brassica campestris [rapeseed] sterois 0.0024

[0051] EXAMPLE 4: PROTECTION OF COLLAGEN XVII BY COMPOSITION ACCORDING TO THE INVENTION

[0052] 4-1 Purpose of the study

[0053] This study aims to determine the protective capacity of the composition according to the invention and corresponding to the mixture of compounds corresponding to the INCI designations Pyrus malus (APPLE) Seed Extract and Brassica campestris sterols and Saccharide Isomerate against the proteolysis of collagen XVII.

[0054] 4-2 Materials and methods

[0055] 4-2.1 - Cell culture

[0056] Normal human keratinocytes were used in this study. The cells were seeded in 96-well plates at a rate of 15,000 cells / well in complete culture medium (KGM). After 24 hours of incubation, the cells were incubated with the compounds of the invention or the positive control (EGF 10 ng / ml). After 48 hours of incubation at 37°C, 5% CO2, the culture supernatants were collected and stored at -80°C before being used for the quantification of cleaved collagen XVII released into the medium by ELIS A. Cell viability was assessed using the MTT (3,4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay in order to normalize the collagen-XVII results to MTT.

[0057] 4-2.2 - Tested Products

[0058] The compounds referenced in Table 4 were tested alone or in combination.

[0059] [Tables4] Active compounds according to the invention (INCI) Dilutions tested: SACCHARIDE ISOMERATE 0.006%, WMALUS (APPLE) SEED EXTRACT & BFLASS / CA CAMPESTRIS STEROLS (Ratio: 0.8) 0.01%

[0060] Since the solubility of Pyrus malus (APPLE) Seed Extract & Brassica campestris sterols is in DMSO, the tested concentrations had been adjusted in 0.1% final DMSO for all tested conditions in order to normalize the effect of the solvent to the results.

[0061] 4-2.3 Quantification of Collagen-17 proteolysis in KHN supernatants

[0062] Quantification of collagen XVII proteolysis was carried out in the following Swimming cells were analyzed by ELISA (Ref.: E-EL-H0785, Elabsciences, USA) by measuring the cleaved collagen XVII released from the cells into the supernatant, according to the manufacturer's instructions. Optical density (OD) was read using a spectrophotometer at 450 nm. Collagen XVII quantification was obtained from the standard curve (0 to 200 ng / ml) established with recombinant human collagen XVII protein. Results were expressed as the percentage of collagen XVII proteolysis normalized to the percentage of viability per well. Collagen XVII assays were performed only when cell viability was greater than 80%.

[0063] 4-2.4 Cell viability using an MTT test

[0064] MTT ((3-4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) is a soluble yellow dye that is metabolized by mitochondrial enzymes (succinate dehydrogenase) into a dark blue compound: formazan. The formazan crystals are then solubilized in DMSO and the optical density is measured by spectrophotometry. trophoshotometer at 540 nm. Cells are treated with 1 mg / ml of MTT for 3 hours and formazan crystals are solubilized in 100 µl of DMSO.

[0065] 4-2.5 Statistical analysis

[0066] Data were collected from independent experiments conducted in triplicate. Quantitative analyses are expressed as mean with standard deviation. Statistical significance is determined by a Student's t-test. Differences are considered statistically significant from p<0.05. (NS: p>0.05; *: p<0.05; **: p<0.01; ***: p<0.001).

[0067] 4.3 Results and conclusions

[0068] The protective effect of the active ingredients of the composition, i.e. Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and saccharide Isomerate against the proteolysis of collagen XVII, was evaluated by the in vitro assay of this protein. The results are shown graphically in [Fig. 1].

[0069] The untreated condition corresponds to the control which arbitrarily represents 100% of proteolysis of collagen-XVII in basal condition.

[0070] The positive control (EGF-10 ng / ml) corresponds to the control of inhibition of collagen XVII proteolysis. It induces a significant decrease in proteolysis of 34%.

[0071] The solvent control (DMSO 0.1%) corresponds to the solvent of the combination of Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and Saccharide Isomerate. It does not exert a significant effect on the synthesis of collagen XVII.

[0072] The active ingredient Saccharide Isomerate induces a significant 41% decrease in collagen-XVII proteolysis at the tested dose, which is significant compared to an untreated or solvent control condition (Student's test; p < 0.005: ***).

[0073] The combination of the active ingredients Pyrus malus (APPLE) Seed Extract and Brassica campestris sterols has no effect on the proteolysis of collagen XVII at the tested dose compared to an untreated or solvent control condition.

[0074] The composition of the invention, comprising the active ingredients Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and Saccharide Isomerate, confers significant and synergistic protection against collagen-XVII proteolysis of 58%, which is significant compared to an untreated or solvent control condition and to a Saccharide Isomerate condition (Student's t-test; 0.05 < p < 0.01: * or #)•

[0075] The synergy between two active substances (in this case: Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols, and Saccharide Isomerate) is calculated by comparing the results of tests in which the efficacy of the two substances and that of their mixture is measured. The Colby formula (Tammes, 1964; Schuldt, PH and Lammont, 1962) is then used.

[0076] If the measured effectiveness of the mixture is greater than that expected according to the Colby formula, there is synergy. If it is less, there is antagonism. And if it is equal, the two substances act without interacting with each other.

[0077] Colby formula: E = X+Y - (XxY) / 100

[0078] X = effect of the active ingredients Pyrus malus (APPLE) Seed Extract and Brassica campestris sterols on the proteolysis of collagen XVII at the tested dose (0.01%).

[0079] Y = effect of the active ingredient Saccharide Isomerate on the proteolysis of collagen XVII at the tested dose (0.006%).

[0080] E = expected efficacy of the active ingredients Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and Saccharide Isomerate at respective doses of 0.01% and 0.006%, without synergy = (0+41) - [(0*41) / 100] = 41%.

[0081] The observed effectiveness of the composition is 58%, i.e. well above 40% which was the expected effectiveness of the composition.

[0082] These data demonstrate the synergistic effect between the active ingredients Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and Saccharide Isomerate in a composition according to the invention.

[0083] The Applicant has demonstrated that the combination of active ingredients Pyrus malus (APPLE) Seed Extract, Brassica campestris sterols and Saccharide Isomerate also allows for the synergistic protection of another type of collagen, collagen I, from glycation.

[0084] Bibliography • Langton, AK, Halai, P., Griffiths, CEM, Sherratt, MJ, and Watson, REB (2016). The impact of intrinsic aging on the protein composition of the dermal-epidermal junction. Mech. Aging Dev. 156, 14-16. • Liu, N., Matsumura, H., Kato, T., Ichinose, S., Takada, A., Namiki, T., Asakawa, K., Morinaga, H., Mohri, Y., De Arcangelis, A., et al. (2019). Stem cell competition orchestrates skin homeostasis and aging. Nature 568, 344-350. • Mondon, P., Hillion, M., Peschard, O., André, N., Marchand, T., Doridot, E., Feuilloley, M.G., Pionneau, C., and Chardonnet, S. (2015). Evaluation of dermal extracellular matrix and epidermal-dermal junction modifications using matrix-assisted laser desorption / ionization mass spectrométrie imaging, in vivo réflectance confocal microscopy, echography, and histology: effect of âge andpepti. J. Cosmet. Dermatol. 14, 152-160. • Nishizawa, Y., Uematsu, J., and Owaribe, K. (1993). HD4, a 180 kDa Bullous Pemphigoid Antigen, Is a Major Transmembrane Glycoprotein of the Hemi-desmosomel. J. Biochem. (Tokyo) 113, 493-501. • Shirai, K., Obara, K., Tohgi, N., Yamazaki, A., Aki, R., Hamada, Y., Arakawa, N., Singh, S.R., Hoffman, R.M., and Amoh, Y. (2019). Expression of anti-aging type-XVII collagen (COL17A1 / BP180) in hair follicle-as-sociated pluripotent (HAP) stem cells during différentiation. Tissue Cell 59, 33-38. Schuldt, PH and Lammont, D. (1962) Application of the method of Limpel LE, PROC. NEWCC 16, 48 - 53. Tammes, PML (1964) Isoboles, a graphical representation of synergism in pesticides.” Netherlands Journal of Plant Pathology 70: 73-80. Watanabe, M., Natsuga, K., Nishie, W., Kobayashi, Y., Donati, G., Suzuki, S., Fujimura, Y., Tsukiyama, T., Ujiie, H., Shinkuma, S., et al. (2017). Type XVII collagen coordinates proliferation in the interfollicular epidermis. ELife 6, e26635.

Claims

Demands

1. Cosmetic composition, advantageously ecobiological, comprising a mixture of an apple seed extract and at least one phytosterol, characterized in that said mixture is combined with at least one exopoly s accaride.

2. Composition according to claim 1 characterized in that the apple seed extract represents between 0.0000001% to 5% by total weight of the composition, advantageously between 0.000001% and 0.3%.

3. Composition according to claim 1 or 2 characterized in that at least one phytosterol, advantageously derived from rapeseed extract, is selected from the group consisting of campesterol, beta-sitosterol, stigmasterol, brassicasterol, campestanol and sitostanol.

4. Composition according to any one of the preceding claims, characterized in that at least one phytosterol represents between 0.0000001% and 5% by total weight of the composition, advantageously between 0.000001% and 0.1%.

5. Composition according to any one of the preceding claims, characterized in that the ratio between the weight of the apple seed extract and the weight of at least one phytosterol is between 0.1 and 2, advantageously between 0.3 and 1.5, preferably between 0.7 and 0.9, more advantageously 0.

8.

6. Composition according to any one of the preceding claims, characterized in that at least one exopolysaccharide is composed by the repetition of a tetrasaccharide consisting of three galacturonic acid units and one N-acetylglucosamine unit in which two of the three galacturonic acid units are substituted with an alanine group and a serine group.

7. Composition according to any one of the preceding claims, characterized in that at least one exopolysaccharide is of bacterial origin, advantageously derived from the marine bacterium Vibrio algi-nolyticus.

8. Composition according to any one of the preceding claims, characterized in that at least one exopolysaccharide represents between 0.0000001% and 5% by total weight of the composition, advantageously between 0.000001% and 0.1%.

9. Composition according to any one of the preceding claims, characterized in that the ratio between the weight of the mixture of the extract of apple seeds and phytosterol and the weight of the exopolysaccharide is between 0.1 and 2, advantageously between 0.3 and 1.5, preferably between 0.5 and 0.

6.

10. Composition according to any one of the preceding claims, characterized in that: - the apple seed extract corresponds to the INCI designation Pyrus malus (APPLE) seed extract; - the phytosterol corresponds to the INCI designation Brassica campestris (Rapeseed) sterols; and / or - the exopolysaccharide corresponds to the INCI designation Saccharide isomerate.

11. Composition according to any one of the preceding claims, characterized in that it further comprises vitamin PP: - advantageously corresponding to the INCI designation Niacinamide, or one of its derivatives; and / or - representing between 0.0001% and 10% by total weight of the composition, advantageously between 0.001% and 5%, preferably between 0.1% and 2%.

12. Composition according to any one of the preceding claims, characterized in that it further comprises - a compound corresponding to the INCI designation hexyldecanol and representing between 0.0000001% and 5% by total weight of the composition, advantageously between 0.000001% and 0.3%; - a compound corresponding to the INCI designation tocopherol and representing between 0.0000000002% and 0.01% by total weight of the composition, advantageously between 0.000000002% and 0.0006%.

13. . Cosmetic use of the composition according to any one of the preceding claims to reduce, or even prevent, the loss of functionality of collagens, advantageously the loss of functionality induced by proteolysis, glycation, acetylation, acylation of collagens, advantageously of collagen I and / or collagen XVII, more advantageously of collagen XVII.

14. Cosmetic use of the composition according to any one of claims 1 to 12 to combat skin aging, advantageously to combat the appearance of wrinkles.

15. Composition according to any one of claims 1 to 12 for use in regenerating skin, advantageously in promoting skin healing.