EXOPOLYSACCHARIDE AND OLIGOSACCHARIDES, ESPECIALLY FOR COSMETICS

Abstract

The present invention relates to an exopolysaccharide comprising N-acetylquinovosamine, oligosaccharides obtainable by cleavage of said exopolysaccharide, a microorganism for their production, a method for obtaining them, compositions containing them, and uses, particularly in the field of cosmetics. The oligosaccharides are especially suitable for improving skin radiance, texture, firmness, and hydration, and for protecting the skin from aging and / or the effects of pollution.

Description

Title of the invention: EXOPOLYSACCHARIDE AND OLIGOSACCHARIDES, PARTICULARLY FOR COSMETICS

[0001] The present invention relates to an exopolysaccharide, oligosaccharides which can be obtained by cleavage of said exopolysaccharide, a microorganism allowing their manufacture, a method of obtaining them, compositions containing them and uses, in particular cosmetic uses.

[0002] The fields of application of the present invention are mainly the chemical industries, for example the food, cosmetic, pharmaceutical, veterinary, agricultural, environmental or paint and coating industries.

[0003] The present invention applies more particularly to the fields of the cosmetics and dermopharmacy industries which manufacture and / or use products intended for the treatment of the skin (including the scalp), mucous membranes and appendages (such as hair, eyelashes, eyebrows, nails, hair) of mammals, animals or humans, to improve their appearance and / or general condition.

[0004] Polysaccharides consist of a more or less long chain of monosaccharides and / or monosaccharide acids linked by glycosidic bonds. They can be linear or branched. They are present in the natural state in plants, mammals or even microorganisms, with the function in particular of energy storage, structure or support of tissues, regulation of the viscoelasticity of tissues, water retention, protection against bacterial or protease attacks and / or blood coagulation.

[0005] The family of polysaccharides contains a particular class consisting of exo-polysaccharides. These are characterized by the fact that they are produced by microorganisms living in ecosystems with extreme conditions and secreted outside the cells.

[0006] One of the advantages of exopolysaccharides for industry is that the microorganisms that produce them can be grown in large volumes under controlled conditions. This makes it easier to produce exopolysaccharides on a large scale compared to polysaccharides, which often require complex extractions from natural sources such as plants or algae.

[0007] Several exopolysaccharides have been described for the field of cosmetics and / or dermatology.

[0008] Patent application US2009 / 069213 describes several exopolysaccharides derived from microalgae strains of the Porphyridium family. An exopolysaccharide derived from the Porphyridium cruentum strain is presented, composed of the three monosaccharides, xylose, glucose and galactose, and monosaccharide glucuronic acid, and recommended for its anti-wrinkle and moisturizing properties.

[0009] Another example is given in patent application WO2012 / 072245 which describes an exopolysaccharide isolated from a strain of Pseudoalteromonas sp. filed under number 1-4150, which is a specifically non-hydrothermal strain. This exopolysaccharide is composed of five monosaccharides, mannose, glucose, galactose, rhamnose and N-acetyl-glucosamine, and two monosaccharide acids, glucuronic and galacturonic. It is recommended as an alternative to hyaluronic acid also for anti-wrinkle and moisturizing effects.

[0010] Yet another example is given in patent application WO2015 / 117985 which describes an exopolysaccharide derived from a culture of Pseudomonas sp. filed under number CECT8437. This exopolysaccharide is composed of three monosaccharides, glucose, galactose and fucose, and a monosaccharide uronic acid. It is recommended for its emulsifying activity in the fields of cosmetics and food.

[0011] Exopolysaccharides can also advantageously be used as a source of saccharide monomers or oligosaccharides through appropriate cutting. For those skilled in the art, oligosaccharides are small polysaccharides often characterized by the fact that they comprise fewer than 25 monosaccharides and / or monosaccharide acids. The advantage of oligosaccharides is that they penetrate the skin barrier more easily and have better stability in formulations due to their smaller size.

[0012] Thus, for example, in the patent application WO2015 / 117985 cited above, the hydrolysis of the exopolysaccharide into oligosaccharides and monosaccharides which can be used in cosmetic or pharmaceutical products is described. No particular activity is presented.

[0013] Another patent application WO98 / 50013 describes the use of oligosaccharides, without specifying their origin. The oligosaccharides described have between two and six osidic residues including a galactose residue in the terminal position. They are recommended in cosmetic compositions to increase the synthesis and / or reduce the degradation of proteoglycans in connective tissue, allowing the skin to be hydrated and to combat the effects of free radicals.

[0014] However, there is still strong commercial demand for new active ingredients that address the current issue of sustainability, particularly offering products derived from biotechnological processes that preserve natural resources.

[0015] The present invention aims in particular to meet this demand.

[0016] To this end, it proposes, according to a first aspect, an exopolysaccharide characterized in that it comprises at least one monosaccharide unit which is the N- acetyl-quinovosamine (NAcQ).

[0017] This new exopolysaccharide advantageously constitutes a source of new oligosaccharides due to the presence of N-acetyl-quinovosamine and having original cosmetic properties as shown by the test results set out later in the description.

[0018] This new exopolysaccharide can also be used advantageously in the technical fields mentioned above, in the native or partially cleaved state. For example, it can be used as a rheological or film-forming agent in formulations. In the field of cosmetics, it is likely to be used to improve the general condition of the skin and appendages, by forming a protective barrier to prevent water loss and create a feeling of softness and comfort. It can also have anti-aging properties via a tightening effect.

[0019] According to other characteristics, the molecular weight of the exopolysaccharide is greater than or equal to 260,000 Da, more particularly greater than or equal to 500,000 Da.

[0020] According to other particular characteristics of the exopolysaccharide according to the invention:

[0021] - it further comprises at least one monosaccharide acid unit chosen from glucuronic acid (Glu) and galacturonic acid (Gal), preferably it comprises these two monosaccharide acid units; and / or

[0022] - it further comprises at least one monosaccharide unit which is rhamnose (Rham); and / or

[0023] - it comprises between 15% and 55% of N-acetyl-quinovosamine, preferably sen possibly 25%; and / or

[0024] - it comprises between 15% and 55% of glucuronic acid, preferably substantially 25%; and / or

[0025] - it comprises between 15% and 55% of galacturonic acid, preferably substantially 25%; and / or

[0026] - it comprises between 15% and 55% of rhamnose, preferably substantially 25%; and / Or

[0027] - it comprises a chain of three saccharide units: glucuronic acid, galacturonic acid and N-acetyl-quinovosamine (Glu-Gal-NAcQ); and / or

[0028] - it also comprises rhamnose, with the following sequence: glucuronic acid, galacturonic acid, N-acetyl-quinovosamine and rhamnose (Glu-Gal-NAcQ-Rham); and / or

[0029] - it consists of the repetition of the sequence glucuronic acid, galactic acid turonic acid, N-acetyl-quinovosamine, rhamnose (Glu-Gal-NAcQ-Rham), each saccharide unit being found in approximately equimolar quantities in the exopolysaccharide.

[0030] All percentages expressed in the above paragraph are by weight relative to the total weight of Texopolysaccharide.

[0031] Furthermore, Texopolysaccharide can be derivatized on the carboxylic acid functions when they are present, in particular Texopolysaccharide can be derivatized so that at least one carboxylic acid end of the glucuronic acid or of the galacturonic acid is for example esterified or acylated.

[0032] According to still other characteristics, Texopolysaccharide is derived from the bacterial strain Pseudoalteromonas sp., deposited by the Applicant at the National Collection of Culture of Microorganisms (CNCM) of Paris under number 1-5893 according to the Budapest Treaty. This bacterial strain was isolated from the surface of a candelabra-shaped coral of the octocoral type belonging to the genus Pleuxaura, collected in the warm waters of the Bimini Islands in the Bahamas. The non-spore-forming bacterium has a rod shape and grows in colonies in salt-rich environments because of its marine origin.

[0033] As mentioned above, Texopolysaccharide according to the invention can be used as a source of oligosaccharides.

[0034] Thus, according to a second aspect, the present invention also provides an oligosaccharide capable of being obtained by cleavage of Texopolysaccharide according to the invention and composed of at least one monosaccharide unit which is N-acetyl-quinovosamine.

[0035] The oligosaccharide according to the invention can also be manufactured synthetically by chemical or enzymatic means, from monosaccharides.

[0036] Advantageously, oligosaccharides according to the invention are presented in the detailed description as having particularly interesting and original cosmetic activities in addition to the classic hydration and anti-wrinkle properties. In particular, the test results show that the oligosaccharides make it possible to combat cellular hypoxia for a very rapid cosmetic benefit on the radiance of the complexion.

[0037] According to particular characteristics of the oligosaccharide according to the invention:

[0038] - has a degree of polymerization between 2 and 12, preferably a degree of polymerization of 3, 4, 7 or 11; and / or

[0039] - further comprises N-acetyl-quinovosamine at least one mono acid unit saccharide chosen from glucuronic acid and galacturonic acid, preferably it comprises these two monosaccharide acid units; and / or

[0040] - includes the chain glucuronic acid, galacturonic acid and N- acetyl-quinovosamine (Glu-Gal-NAcQ); and / or

[0041] - further comprises rhamnose with the following chain: glucuronic acid, galacturonic acid, N-acetyl-quinovosamine, rhamnose (Glu-Gal-NAcQ-Rham).

[0042] According to other characteristics, the average molecular weight of a saccharide unit of the exopolysaccharide according to the invention being approximately 200 Da (corresponding to the average of the mass of a unit of N-acetyl-quinovosamine, glucuronic acid, galacturonic acid and rhamnose), the oligosaccharides according to the invention preferably have a molecular weight less than or equal to 5000 Da corresponding to a degree of polymerization of 25, more preferably a molecular weight less than or equal to 3000 Da corresponding to a degree of polymerization of 15, more preferably less than or equal to 2200 Da corresponding to a degree of polymerization of 11, more preferably less than or equal to 1400 Da corresponding to a degree of polymerization of 7, more preferably less than or equal to 800 Da for a degree of polymerization less than or equal to 4, more preferably still less than or equal to 600 Da for a degree of polymerization less than or equal to 3,and even more preferably less than or equal to 400 Da for a degree of polymerization less than or equal to 2.,

[0043] The preferred oligosaccharides according to the invention are: - a trisaccharide with a degree of polymerization of 3, comprising an N-acetyl-quinovosamine unit, a glucuronic acid unit and a galacturonic acid unit and having a molecular weight of 557 Da; more particularly consisting of the Glu-Gal-NAcQ chain; and / or - a heptasaccharide with a degree of polymerization of 7, comprising two N-acetyl-quinovosamine units, two glucuronic acid units, two galacturonic acid units and one rhamnose unit and having a molecular weight of 1242 Da; more particularly consisting of the Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ chain; and / or - a hendecasaccharide with a degree of polymerization of 11, comprising three N-acetyl-quinovosamine units, three glucuronic acid units, three galacturonic acid units and two rhamnose units and having a molecular weight of 1927 Da, more particularly consisting of the chain Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ.

[0044] Furthermore, the oligosaccharide according to the invention can be derivatized on the carboxylic acid functions when they are present, in particular the oligosaccharide can be derivatized so that at least one carboxylic acid end of the glucuronic acid or of the galacturonic acid is for example esterified or acylated.

[0045] As presented above, the exopolysaccharide according to the invention and consequently the oligosaccharides resulting therefrom can be obtained by a fermentation process, in particular from strain 1-5893 deposited at the CNCM.

[0046] According to a third aspect, the present invention therefore provides a bacterial strain Pseudoalteromonas sp., deposited by the Applicant in the Collection National Institute of Microorganism Culture (CNCM) of Paris under number 1-5893.

[0047] According to a fourth aspect, the present invention provides a manufacturing method by culturing this strain to produce the exopolysaccharide according to the first aspect and oligosaccharides according to the second aspect, said method comprising the following steps: - a first step of culturing a bacterial strain of the genus Pseudoalteromonas sp. 1-5893 deposited at the CNCM in a culture medium adapted to the production of the bacteria excreting the exopolysaccharide in the culture medium; and - a second step of recovery of the exopolysaccharide separated from the culture medium.

[0048] Advantageously, in the first step, the exopolysaccharide is excreted by the bacterial strain into the culture medium at the same time as the amount of bacterial strain is increased.

[0049] The culture medium is liquid and may contain macroelements, microelements, yeast extracts, vitamins and sugars. It is selected to promote the growth and production of the exopolysaccharide according to the invention.

[0050] As microelements, mineral salts can be used for the culture. For example and not limited to, salts providing the ions Na+, K+, NH4+, Ca2+, Mg2+, PO43, SO42, Cl and / or CO32, or trace elements such as Cu, Mn, Fe and / or Zn, or a mixture thereof. These salts are selected to promote the growth and production of the exopolysaccharide.

[0051] Similarly, sugars may be used as a carbon source to feed the bacterial strain such as, but not limited to, galactose, glucose, mannose, amygdalin, cellobiose, maltose, starch, glycogen, lactose or a mixture thereof. The addition of sugars also increases the production of the exopolysaccharide.

[0052] According to other characteristics, in the second step, the separation of the exopolysaccharide from the culture medium is done in two stages.

[0053] Firstly, the exopolysaccharide present in the liquid culture medium is separated from the biomass which is solid, by a process for separating a liquid phase from a solid. For example, without limitation, the separation can be carried out by centrifugation, ultrafiltration, tangential filtration, or press filtration. The biomass contained in the solid phase can be removed or retained to be added, after extraction, to the exopolysaccharide obtained at the end of the process in order to enrich it with proteins and sugars which are the compounds present in the biomass.

[0054] In a second step, the exopolysaccharide is purified by isolating it from the liquid culture medium containing other molecules which can be salts, proteins, etc. For example, these molecules can be precipitated by reducing the temperature, called cold precipitation, or by using a water-miscible solvent in which the exopolysaccharide is insoluble, for example methanol, isopropanol, ethanol or a mixture of these, or by carrying out tangential filtration.

[0055] Preferably, the exopolysaccharide is isolated from the culture medium by tangential filtration. More particularly, a filter membrane of a size smaller than the exopolysaccharide is used. Even more particularly, the filter membrane has a size less than 150,000 Da.

[0056] In an additional step, the exopolysaccharide obtained by the above method can be cleaved to obtain at least one oligosaccharide according to the invention. This cleavage can be carried out by chemical or enzymatic hydrolysis.

[0057] Chemical hydrolysis in an acidic medium can be carried out by a strong acid, such as hydrochloric acid, nitric acid, sulfuric acid or trifluoroacetic acid.

[0058] Enzymatic hydrolysis can be carried out by a specific hydrolase-type enzyme.

[0059] For these different types of hydrolysis, the concentration of the solvent or enzyme, the reaction time and the temperature are defined according to the size of the oligosaccharides that one wishes to obtain. For example, the higher the acid concentration or the longer the reaction time, the smaller the oligosaccharides that one will obtain.

[0060] Acid hydrolysis is the preferred method and the solvent preferably used is hydrochloric acid at a concentration of 0.2 M.

[0061] During hydrolysis, precipitates may appear in the liquid medium containing the oligosaccharides. These precipitates may consist of salts, proteins, cellular debris, etc. which are to be removed in order to purify the liquid phase. To do this, any method is used which makes it possible to separate a liquid phase from a solid. For example, the separation may be carried out by centrifugation, ultrafiltration, tangential filtration or press filtration.

[0062] In the exopolysaccharide according to the invention, the chemical bonds around the rhamnose are cleaved because they are the most fragile. The exopolysaccharide having the repetitive sequence glucuronic acid, galacturonic acid, N-acetyl-quinovosamine and rhamnose, the following oligosaccharides can in particular be obtained mainly:

[0063] - trisaccharides with glucuronic acid, galactoic acid as the chain turonic acid and N-acetyl-quinovosamine (Glu-Gal-NAcQ);

[0064] - other oligosaccharides with a chain of trisaccharides with a rhamnose between each trisaccharide, such as in particular:

[0065] - heptasaccharides corresponding to two Glu-Gal-NAcQ trisaccharides linked by a rhamnose (Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ);

[0066] - hendecasaccharides corresponding to three linked Glu-Gal-NAcQ trisaccharides each by a rhamnose (Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ).

[0067] According to a fifth aspect, the present invention provides a mixture of oligosaccharides obtainable according to the method of the fourth aspect of the invention, preferably composed mainly of trisaccharides and / or heptasaccharides according to the invention, i.e. more than 50%, more particularly more than 80%. The percentages are expressed by weight relative to the total weight of the mixture of oligosaccharides.

[0068] Each oligosaccharide according to the invention can be isolated from the mixture of oligosaccharides obtained by the process according to the invention by an appropriate technique, for example by flash chromatography on silica.

[0069] The exopolysaccharide and the oligosaccharides according to the invention, in particular obtained by the process described above, can be used in cosmetic or dermatological compositions as active ingredients.

[0070] Thus, according to a sixth aspect, the present invention provides a cosmetic or dermatological composition comprising the exopolysaccharide according to the invention as an active ingredient and a physiologically acceptable medium.

[0071] Thus also, according to a seventh aspect, the present invention provides a cosmetic or dermatological composition comprising at least one oligosaccharide according to the invention as active ingredient according to the invention and a physiologically acceptable medium.

[0072] By "physiologically acceptable medium" is meant according to the present invention, without being limiting, an aqueous or hydroalcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion, an aqueous gel, an anhydrous gel, a serum, a dispersion of vesicles, a powder.

[0073] “Physiologically acceptable” means that the compositions are suitable for a topical use in contact with mucous membranes, nails, scalp, hair, fur and skin of mammals and more particularly humans, without risk of toxicity, incompatibility, instability, allergic response, and others.

[0074] This “physiologically acceptable medium” forms what is classically called the excipient of the composition.

[0075] Preferably according to the invention, the physiologically acceptable medium may be an aqueous, hydroglycolic or hydroalcoholic medium, or formed by a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion or a mixture thereof. More preferably, it is a mixture of water, glycol and diol.

[0076] According to other advantageous characteristics, the exopolysaccharide and / or the at least one oligosaccharide may be used in combination with one or more additional active ingredients at effective concentrations advantageously providing a synergistic or reinforcing effect to achieve the desired effects described for the invention.The additional active ingredients may be chosen, for example, from lightening, anti-redness, anti-blemish, calming active ingredients for the treatment of sensitive, reactive skin, filtering radiation, in particular UVA, UVB, IR, from blue light, moisturizing, humectant, exfoliating, smoothing, toning, anti-aging, anti-wrinkle and fine lines active ingredients, improving mechanical and elastic properties, radiance of the complexion, detoxifying, anti-hair regrowth active ingredients, acting on the skin barrier, anti-acne, acting on sebum secretion, mattifying, unifying, anti-inflammatory, antioxidant, anti-free radical, anti-glycating, eye contour (anti-eye bags and anti-puffiness), promoting blood circulation, regenerating, acting favorably or unfavorably on the skin microbiota and that of its appendages, promoting the growth of hair and / or hair, promoting the elimination of toxins, etc.These active ingredients can be obtained from plant materials, such as plant extracts or products of plant culture or fermentation.

[0077] More specifically, the exopolysaccharide and / or the at least one oligosaccharide according to the invention may also be combined with at least one of the compounds chosen from vitamin B3 compounds, compounds such as niacinamide or tocopherol, retinoid compounds such as retinol, in particular in an encapsulated form, hexamidine, α-lipoic acid, resveratrol or DHEA, hyaluronic acid, ceramides or ceramide analogues, obtained synthetically or by biotechnological means, peptides, in particular N-acetyl-Tyr-Arg-O-hexadecyl, VW, Pal-PP, Biot-GHK, Ela-KFK, Pal-KPK, Pal-KHG, Myr-SPA, Pal-VGVAPG (SEQ ID NO: 1), Pal-KTTKS (SEQ ID NO: 2), Pal-GHK, Pal-KM02K, Pal-GQPR (SEQ ID NO: 3), Pal-K(Ac)HG, Pal-K(P)HG and Pal-KTSKS (SEQ ID NO: 4), Pal-KTFK (SEQ ID NO: 5), Pal-YGGFL (SEQ ID NO: 6), cyclolinopeptides, plant extracts, in particular an extract of Marrubium vulgare,an extract of Leontopodium Alpinum produced by in vitro cell culture, an extract of Mirabilis jalapa, or mixtures, in particular of an extract of Rosmarinus officinalis and Pal-GQPR (SEQ ID NO: 3), which are known active ingredients that can be used in topical cosmetic or dermo-pharmaceutical compositions.

[0078] According to an eighth aspect, the present invention proposes the use of the exopolysaccharide and / or at least one oligosaccharide and / or one of the compositions containing it according to the different aspects of the invention in the industrial fields mentioned above, namely mainly the chemical industries, for example the food, cosmetic, pharmaceutical, veterinary, agricultural industries, environmental or paints and coatings.

[0079] More particularly, the present invention proposes the use of at least one oligosaccharide and / or the composition containing it according to the different aspects of the invention for a non-therapeutic cosmetic treatment of the skin and its appendages. Preferably according to the invention, the treatment is topical.

[0080] Preferably, the treatment is carried out with a mixture of oligosaccharides according to the fifth aspect of the invention.

[0081] According to the invention, by “topical treatment” or “topical use” is meant an application which is intended to act at the place where it is applied: skin, mucous membrane and / or appendages.

[0082] More particularly, the cosmetic treatment proposed by the present invention seeks to combat the unsightly signs of skin damage resulting from the loss of cell vitality and the loss or reduction of certain essential functions of the skin.

[0083] Loss of radiance, firmness, hydration and / or the appearance of roughness of the skin are among these unsightly signs. They are due to normal, chronological aging of the skin and / or premature aging resulting from extrinsic factors such as external aggressions, in particular atmospheric pollutants or radiation. Over time and / or in the presence of these extrinsic factors, the vital functions of the skin cells as well as certain functions at the skin level will deteriorate.

[0084] According to particular characteristics, the cosmetic treatment using at least one oligosaccharide according to the invention is suitable for:

[0085] - improve the radiance of the complexion, also called radiance;

[0086] - improve the relief of the skin;

[0087] - improve skin firmness; and / or

[0088] - improve skin hydration.

[0089] Indeed, the tests carried out in vitro and presented later in the detailed description have shown that the treatment according to the invention based on oligosaccharides advantageously allows:

[0090] - to act at the level of skin cells: by fighting against oxidation and hypoxia cellular by stimulating energy production, oxygen consumption and cellular respiration; and

[0091] - to act at the skin level: by strengthening the barrier function, the hy deration, dermoepidermal junction (DEJ) function and microbiota function.

[0092] More specifically at the cell level:

[0093] The tests showed that the treatment according to the invention confers a beneficial antioxidant effect by reducing the synthesis of reactive oxygen species (ROS) thanks to to the increase in the synthesis of antioxidant proteins encoded by the Nrf2, NQO-1 and HMOX-1 genes.

[0094] During oxidative stress, the protein encoded by the Nrf2 gene enters the cell nucleus and induces the activity of certain genes such as NQO-1 and HMOX-1.

[0095] NQO-1 is a gene encoding an antioxidant protein that serves to maintain a balance of the intracellular oxidative state and also to detoxify aromatic pollutants. This protein is synthesized when ROS increases in the cell.

[0096] HMOX-1 is the gene encoding the protein Heme-oxygenase (HO-1) which is induced rapidly after exposure to free radicals. HMOX-1 has a protective role against oxidative stress mainly due to its ability to degrade Heme into carbon monoxide and biliverdin, a precursor of bilirubin which is a powerful antioxidant.

[0097] Thanks to this antioxidant action, skin aging is limited, in particular by preserving the dermal extracellular matrix (ECM). Indeed, oxidation negatively affects the synthesis of molecules, such as fibroblasts, which are responsible for the synthesis of collagen, elastin and other components of the ECM that help maintain the structure by preventing wrinkles and fine lines, maintaining firmness and maintaining the elasticity of the skin.

[0098] The tests showed that the treatment according to the invention acted on cellular hypoxia:

[0099] - mitochondrial potential creating energy in cells that need it to make proteins, operate enzymes, detoxify cells from accumulated pollutants, moisturize the skin, etc., is increased;

[0100] - oxygen consumption also allows energy to be created. Indeed, oxygen is consumed to allow CO2 to be evacuated and energy to be produced;

[0101] - the production of adenosine triphosphate (ATP) which activates cellular respiration in stimulating detoxification mechanisms and increasing cell production, is increased.

[0102] More energy and oxygenation of the cells results in detoxification and regeneration of the cells, thus improving the quality of the epidermis and strengthening the skin barrier for a better radiance of the complexion.

[0103] More specifically at the skin level:

[0104] In vitro tests have shown that the treatment according to the invention acts on the reinforcement of the skin barrier by acting on the quality and thickness of the stratum corneum.

[0105] The stratum corneum or horny layer is the outermost layer of the epidermis and constitutes a physical and functional barrier with the role in one sense of preventing the penetration of harmful external agents, such as allergens, pathogens and pollutants and in the other direction to avoid dehydration of the skin.

[0106] The stratum corneum is composed of corneocytes, lipids, lamellar bodies and natural moisturizing factors (NMFs) - substances capable of fixing water in the stratum corneum) which maintain the degree of hydration of the skin at an optimal level.

[0107] Transepidermal Water Loss (TEWL) characterizes the evaporation of water contained in the skin: the more the skin barrier is damaged, the more easily the water evaporates.

[0108] Better differentiation of keratinocytes leads to a strengthening of the skin barrier and therefore contributes to better protection of the epidermis and better hydration.

[0109] Keratinocytes are viable cells of the epidermis with a lipid border that migrate from the deepest layer to the most superficial layer, differentiating into corneocytes, which are small flat bricks without a nucleus. Proteins such as involucrin, filaggrin, and claudin-1 are involved in the formation and migration of keratinocytes into corneocytes. Furthermore, corneocytes are strongly linked together by corneodesmosomes, the main protein of which is corneodesmosine, and "cemented" by organized layers of complex extracellular lipids: cholesterol, ceramides, and neutral lipids.

[0110] Involucrin is a protein synthesized in the cytoplasm of keratinocytes in the stratum granulosum. By binding to the protein loricrin, involucrin contributes to the formation of a cell envelope that protects the corneocytes of the skin.

[0111] Claudin-1 protein is a constituent of the tight junctions of the stratum granulosum, which form a selective barrier controlling the paracellular transport of inflammatory molecules and cells. These tight junctions are also essential for limiting water loss.

[0112] Tests showed that these essential proteins were affected: involucrin, comeodesmosin and claudin-1.

[0113] The tests also showed that the treatment according to the invention acted on the hydration function of the skin, by further stimulating the synthesis of aquaporin-3 (AQP-3) which is a known marker of hydration and which is present in the epidermis as well as filaggrin which plays a pivotal role in maintaining the barrier function and whose degradation products are essential for the regulation of pH and cutaneous hydration. Consequently, advantageously, thanks to the treatment according to the invention, not only is dehydration of the skin avoided with a reinforced skin barrier, but also the hydration function itself is stimulated by increasing the synthesis of one of the macromolecules responsible for hydration in the epidermis. Better hydrated skin is plumper, more supple and softer (less rough).

[0114] Thanks to the treatment according to the invention, the skin will be better protected and better hydrated, and therefore more radiant.

[0115] The treatment according to the invention also makes it possible to act on the function of the DEJ by ensuring good anchoring of the keratinocytes to the basal lamina and better communication between the cells allowing polarization of the keratinocytes. The synthesis of the main components of the DEJ (collagen IV, collagen VII and laminins) decreases during aging, which has significant repercussions on the firmness of the skin.

[0116] Tests have shown that the treatment according to the invention acts on markers of the DEJ, namely collagen IV and collagen VII to improve skin firmness. These molecules also participate in strengthening the skin barrier.

[0117] The treatment according to the invention also makes it possible to act on the cutaneous microbiota function.

[0118] This function is ensured mainly by the presence of the Staphylococcus epidermidis strain on the surface of the skin. It is a skin-colonizing germ that is known for its beneficial effects on cutaneous immunity. In addition, Staphylococcus epidermidis has been shown to promote barrier function homeostasis through its ability to cleave sphingomyelin 1) into phosphocholine, which serves as a nutrient substrate and allows it to colonize the skin space, and 2) into type 2 ceramide, which promotes a quality skin barrier.

[0119] The tests showed that the treatment according to the invention acted on the growth of Staphylococcus epidermidis.

[0120] All these effects will thus advantageously and synergistically help to combat dull complexion (loss of radiance), loss of firmness and lack of hydration of the skin, which will help to combat unsightly signs of skin damage.

[0121] Furthermore, advantageously, the treatment according to the invention is particularly suitable for combating pollution, in particular atmospheric pollution.

[0122] Results of specific tests on cellular hypoxia and cell detoxification are given later in the detailed description, carried out in the presence of a solution of atmospheric urban microparticles (UMP) to simulate atmospheric pollutants.

[0123] In vivo tests were carried out confirming the cosmetic benefits according to the invention, in particular a test on the radiance of the complexion (or radiance), a test on the microrelief demonstrating a smoothing effect, a hydration test and a firmness test.

[0124] The present invention therefore covers a method of cosmetic, non-therapeutic topical treatment for beautifying or improving the appearance and general condition of the skin and / or appendages and treating imperfections, in a subject who needs an effective amount of oligosaccharide and / or mixture of oligosaccharides according to the invention or of compositions comprising them in a physiologically acceptable excipient.

[0125] The "effective" amount of the oligosaccharide and / or mixture of oligosaccharides according to different aspects of the invention, i.e. its dosage, depends on the destination of the treatment. It depends on various factors, such as age, condition of the patient, severity of the disorder. An effective amount means a non-toxic amount sufficient to obtain the desired effect.

[0126] In a composition according to the invention, the at least one oligosaccharide, to be present in an effective amount, is generally found in proportions of between 0.000001% and 15% relative to the total weight of the composition, more preferably between 0.00001% and 10%, depending on the intended use of the composition and the desired more or less pronounced effect. More preferably, the effective amount is between 0.0001% and 5% relative to the total weight of the composition.

[0127] According to other particularities, the cosmetic treatment method according to the invention can be associated with one or more other treatment methods targeting the skin, such as for example treatments by light therapy, by heat or by aromatherapy.

[0128] According to the invention, it is possible to propose multi-compartment devices or kits intended for implementing the method described above, and which could comprise, by way of example, and without this being limiting, in a first compartment a composition containing the at least one oligosaccharide according to the invention in an excipient, and in a second compartment a composition containing an additional active agent, the compositions contained in said first and second compartments being considered here as a combination composition for simultaneous, separate or spread-out use over time, in particular in one of the treatments defined above.

[0129] For example, for a cosmetic facial treatment, the European Cosmetics Directive has set a standard application quantity of 2.72 mg / cm2 / day / person for a cream and 0.5 mg / cm2 / day / person for a body lotion.

[0130] The present invention also relates to the use of the exopolysaccharide and / or the oligosaccharide and / or the mixture of oligosaccharides according to the invention, for the manufacture of compositions for a cosmetic treatment, as also described above.

[0131] The composition for use according to the invention can be provided in any galenic form (examples are given later in the description) defined according to the destination of the composition and its site of application.

[0132] A composition according to the invention can be applied to the face, body, décolleté, in any form or vehicle known to those skilled in the art, in particular in the form of a solution, dispersion, emulsion, paste or powder, individually or in premixture or be carried individually or in premixture by vectors such as macrocapsules, microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macrosponges, microsponges or nanosponges, microemulsions or nanoemulsions, or adsorbed on powdery organic polymers, talcs, bentonites, spores or exines and other mineral or organic supports.

[0133] In cosmetics in particular, applications can be proposed, for example in facial and / or body skin care ranges and makeup care ranges, in particular eyelashes and eyebrows.

[0134] For example, the galenic form of the composition may be a lotion, a cream, a butter, a milk, a solid form, a mousse, a gel, a deodorant, an antiperspirant, a shampoo, a conditioner, a hair mask, a face mask, a shower gel, etc.

[0135] The galenic formulas can be included in ranges of personal care products and / or beauty products, in particular ranges of skin care, cleansing, makeup, makeup removal, anti-sun, artificial tanning, pre-shave, shaving, or after-shave, moisturizer, humectant, emollient, conditioner, exfoliant, astringent, depilatory, anti-perspirant or anti-perspirant, deodorant, deodorant, etc.

[0136] The composition may also be incorporated on a non-woven or woven material, made of natural or synthetic fibers, wool, or on any material intended to come into contact with the skin and which may be used in clothing, in particular tights and socks, shorts, day or night underwear, handkerchiefs or fabrics, in order to exert its cosmetic effect via this skin / textile contact and allow continuous topical delivery (cosmetotextiles).

[0137] According to particularly interesting characteristics, the cosmetic treatment using oligosaccharides according to the invention is suitable for:

[0138] - improve the radiance of the complexion;

[0139] - improve the relief of the skin;

[0140] - improve skin firmness; and / or

[0141] - improve skin hydration.

[0142] More particularly, the cosmetic treatment may be a treatment which protects the skin:

[0143] - aging; and / or

[0144] - pollution, particularly atmospheric.

[0145] More particularly still, the treatment:

[0146] - improves skin quality by strengthening the skin barrier for more firmer, more supple, smoother, more hydrated and therefore a better radiance of the complexion; and / or

[0147] - fight against cellular hypoxia for cells for a very cosmetic benefit quick on the radiance of the complexion.

[0148] These cosmetic effects can be envisaged according to the invention separately or in combination, advantageously making it possible in particular to propose a use combining beautifying and sensory effects. DETAILED DESCRIPTION

[0149] The present invention will be better understood in light of the detailed description of exemplary embodiments, studies and figures described below.

[0150] [Fig-1]: it represents a graph of deformation of the skin under the effect of a deformation mechanical training as a function of time.

[0151] 1. Example of production of exopolysaccharide from strain 1-5893

[0152] First step: production of the exopolysaccharide.

[0153] The Pseudoalteromonas sp strain number 1-5893 is inoculated into a bioreactor containing a culture medium for a time sufficient to allow the quantity of strain to multiply.

[0154] The culture medium is then transferred to a larger bioreactor and fresh culture medium can be added to increase strain production. Incubation is carried out for a sufficient time to obtain an even greater quantity of the Pseudoalteromonas sp. strain. Optionally, this step is repeated.

[0155] The culture medium used comprises: - from 0.1% to 0.5% yeast extract; from 0.1% to 0.5% KH2PO4; from 0.1% to 0.5% Na2HPO4; from 0.1% to 0.5% of NH4C1; from 0.001% to 0.01% KC1; - from 0.001% to 0.01% iron citrate; - at least 0.05% anti-foam; and - from 1% to 5% sea salts.

[0156] Glucose, a carbon source, between 1% and 5%, is also added to the culture medium to increase the production of the exopolysaccharide.

[0157] Optionally, fresh culture medium may be added to the bioreactor during incubation and / or minerals and / or sugars during incubation.

[0158] The culture under these conditions is left for a sufficient time for the strain to excrete the exopolysaccharide according to the invention into the culture medium and obtain the desired quantity.

[0159] The incubation is preferably carried out between 25 and 35°C, for example 30°C, the pH between 6.5 and 8 and the dissolved oxygen pressure (pO2) between 10% and 50% by increasing the agitation. When the bacteria are in the exponential phase, the pO2 is regulated in order to stimulate the excretion of the exopolysaccharide into the medium.

[0160] During incubation, the bacterial strain excretes the exopolysaccharide into the culture medium.

[0161] Second step: Recovery of the exopolysaccharide.

[0162] Once the incubation period is over, the culture medium containing the exopolysaccharide (liquid) is separated from the biomass (solid). To separate the liquid phase from the solid phase, continuous centrifugation at a speed of 9000 g is used.

[0163] The biomass contained in the solid phase can either be eliminated or be preserved to be added, after extraction, to the exopolysaccharide obtained at the end of the process according to the invention or added to the mixture of oligosaccharides.

[0164] Once the liquid phase containing the exopolysaccharide has been recovered, the exopolysaccharide according to the invention is isolated from the other molecules contained in the liquid phase. Tangential filtration is used with a 100,000 Da membrane.

[0165] Tangential filtration allows the molecules of a liquid to be separated by their size: molecules with a size less than 100,000 Da pass through the membrane to be eliminated. Since exopolysaccharide has a size greater than 260,000 Da, it does not pass through the membrane and can therefore be recovered.

[0166] This gives the exopolysaccharide washed of undesirable molecules and concentrated.

[0167] Optionally, the exopolysaccharide obtained can then undergo different types of treatment: dilution, preservation, purification, fractionation by precipitation, by chromatography, lyophilization, zeodration or atomization. Characterization of the exopolysaccharide

[0168] During fermentation, secretion of the exopolysaccharide can be monitored by aqueous size exclusion chromatography (SEC) with evaporative light scattering detector (ELSD) detection.

[0169] The obtained exopolysaccharide was characterized by Nuclear Magnetic Resonance (NMR) spectrometry and direct infusion into an ORBITRAP high-resolution (HR) mass spectrometer.

[0170] Cross-analysis of the spectra made it possible to determine the repetitive sequence that makes up the exopolysaccharide: Glu-Gal-NAcQ-Rham.

[0171] 2. Example of manufacturing the mixture of oligosaccharides from 1 ' exopolysaccharide

[0172] The exopolysaccharide according to the invention obtained using the method described above in point 1. can be cleaved to obtain oligosaccharides.

[0173] The cleavage is carried out by an acid hydrolysis step. The exopolysaccharide is mixed with a 0.2 M concentrated hydrochloric acid solution with a pH <l. Le tout est chauffé à une température comprise entre 85°C et 95°C, par exemple 90°C, pendant une durée comprise entre 2 heures et 4h30.

[0174] To stop the hydrolysis, a NaOH solution is used to increase the pH of the mixture, for example to pH 3.

[0175] During the hydrolysis step, precipitates appear in the liquid medium containing the oligosaccharides. A clarification step is therefore carried out. They can be removed by centrifugation, ultrafiltration, tangential filtration or press filtration.

[0176] A mixture of oligosaccharides is obtained comprising mainly trisaccharides and / or heptasaccharides according to the invention.

[0177] Optionally, each oligosaccharide according to the invention can be separated from the rest of the mixture of oligosaccharides by flash chromatography on silica.

[0178] Characterization of the mixture of oligosaccharides obtained

[0179] The oligosaccharides present in the mixture have a degree of polymerization greater than 2 and less than 12 and correspond to assemblies of the saccharide units of the exopolysaccharides, such as N-acetyl-quinovosamine, galacturonic acid and glucuronic acid and where appropriate rhamnose.

[0180] The mixture of oligosaccharides comprises approximately 85% of oligosaccharides having the sequence Glu-Gal-NAcQ, predominantly once the sequence which corresponds to trisaccharides and / or twice the sequence which corresponds to heptasaccharides.

[0181] The majority sequence Glu-Gal-NAcQ present in the oligosaccharide mixture was characterized by NMR spectrometry and by ultra-high performance liquid chromatography-high resolution mass spectrometry ORBITRAP.

[0182] Cross-analysis of the spectra made it possible to determine:

[0183] - that the structure of oligosaccharides is a chain of units of exopolysaccharide: glucuronic acid, galacturonic acid, N-acetyl-quinovosamine and where appropriate rhamnose.

[0184] - that the majority oligosaccharides of the oligosaccharide mixture are:

[0185] - trisaccharides having the following chain: Glu-Gal-NAcQ; and / or

[0186] - heptasaccharides having the following chain: Glu- Gal-NAcQ-Rham-Glu-Gal-NAcQ;

[0187] - that the mixture of oligosaccharides comprises other oligosaccharides in quantity minorities such as:

[0188] - tetrasaccharides having the following chain: Glu-Gal-NAcQ-Rham;

[0189] - hendecasaccharides having the following chain: Glu- Gal-NAcQ-Rham-Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ.

[0190] 3. Example of manufacturing an active ingredient, particularly for cosmetics containing the mixture of oligosaccharides

[0191] The mixture of oligosaccharides according to the invention obtained in point 2. above, is solubilized in a physiologically acceptable medium comprising water, humectants and preservation boosters such as pentylene glycol and propanediol, so as to have a concentration of oligosaccharides of between 4000 ppm and 7000 ppm, for example 5000 ppm.

[0192] 4. Examples of compositions containing the active ingredient

[0193] Different cosmetic formulations are described below using the active ingredient according to the invention as described above in point 3. Additional active ingredients, where appropriate supporting and / or complementing the activity, may be added in the appropriate phase according to their hydrophobic, hydrophilic or amphiphilic nature. These ingredients may be of any category, for example ingredients of telluric origin, benthic, marine, aerial, plant or microbial origin such as bacteria, archaea, or fungi. The ingredients are selected according to their function(s), the place of application (body, face, neck, bust, hands, hair, etc.), the desired final effect and the targeted consumer, for example specific anti-wrinkle, moisturizing, anti-dark circles, firming, anti-glycation, volumizing, soothing, muscle relaxant, anti-redness, detoxifying, etc.

[0194] In the case of the present invention, the following additional active ingredients marketed by Sederma are particularly advantageous: - to reinforce the activity of the active ingredient according to the invention such as BB-BIONT™ for radiance of the complexion and smoothing of the skin, CITYSTEM™, PACIFEEL™ and / or SYNCHROLIFE™ for their anti-pollution activity, AQUALANCE™ for its moisturizing action, CERAMIDE 2™, CERAMIDE A2™, CERAMIDE HO3™ and / or one or more products from the DS-CERAMIDE™ range for their skin barrier reinforcement activity; - in addition to the activity of the active ingredient according to the invention such as MEL[O]STEM™ for its anti-white and brown spot activity, CAL-MOSENSINE™ and / or PACIFEEL™ for their skin soothing activity, MAJESTEM™ DRYPURE™ for its anti-aging activity, one or more products from the MATRIXYL™ range in particular for their activity on the dermal ECM.

[0195] The "International cosmetic ingredient dictionary & handbook" published by "the Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, DC describes a wide variety, without limitation, of cosmetic and pharmaceutical ingredients commonly used in the skin care industry, which are suitable for use as additional ingredients in the compositions according to the present invention, as long as they are physically and chemically compatible with the other ingredients of the composition and especially with the active ingredients of the present invention. Furthermore, the nature of these additional ingredients must not unacceptably alter the benefits of the active ingredients of the invention. These additional ingredients may be synthetic or natural, such as plant extracts or come from a biofermentation process.

[0196] Other skin care active ingredients which are particularly useful in combination with the composition according to the invention can be found in the commercial documentation of Crodarom and Sederma, and on the website www.croda.fr.

[0197] Examples of commercial active ingredients include: betaine, glycerol, Actimoist Bio 2™ (Active organics), AquaCacteen™ (Mibelle AG Cosmetics), Aquaphyline™ (Silab), AquaregulK™ (Solabia), Carciline™ (Greentech), Codiavelane™ (Biotech Marine), Dermaflux™ (Arch Chemicals, Inc), Hydra'Flow™ (Sochibo), Hydromoist L™ (Symrise), RenovHyal™ (Soliance), Seamoss™ (Biotech Marine), Argireline™ (trade name for Lipotec's acetyl hexapeptide-3), spilanthol or an extract of Acmella oleracea known as Gatuline Expression™, an extract of Boswellia serrata known as Boswellin™, Deepaline PVB™ (Seppic), Syn-AKE™ (Pentapharm), Ameliox™, Bioxilift™ (Silab), Phy-toCellTec™Argan (Mibelle), Papilactyl D™ (Silab), Preventhelia™ (Lipotec), or one or more of the following active ingredients sold by Sederma: Subliskin™, Venuceane™, Moist 24™, Vegesome Moist 24™, Essenskin™, Juvinity™, Revidrat™, Resistem™, Chronodyn™, Kombuchka™, Chromocare™,Cal-mosensine™, Glycokin factor S™, Biobustyl™, Idealift™, Ceramide 2™, Ceramide A2™, Ceramide HO3™, Legance™, Intenslim™, Prodizia™, Beautifeye™, Pa-cifeelTM, Zingerslim™, Meiritage™, Sebuless™, Apiscalp™, Rubistem™, Citystem™, Neonyca™, NG Unsaponifiables Shea Butter™, Majestem™, Hydronesis™, Poretect™, Amberstem™, Synchrolife™, Sylverfree™, Feminage™, Ameyezing™, BB-Biont™, Revitalide™, Mel[o]stem™ or mixtures thereof. ,

[0198] Among the plant extracts (in the form of conventional extracts or prepared by an in vitro method) which can be used as additional active agents, mention may be made, in addition and in particular, of ivy extracts, for example climbing ivy (Hedera helix), Bupleurum chinensis, Bupleurum falcatum, arnica (Arnica montana L.), rosemary (Rosmarinus officinalis N.), marigold (Calendula officinalis), sage (Salvia officinalis L.), ginseng (Panax ginseng), ginko biloba, St. John's wort (Hyperycumperforation), butcher's broom (Ruscus aculeatus L.), ulm (Filipendula ulmaria L.), orthosiphon (Orthosiphon stamincus Benth.), artichoke (Cynara scolymus), algae (Fucus vesiculosus), birch (Betula alba), green tea, kola nut (Cola nipida), horse chestnut, bamboo, Centella asiatica, heather, fucus, willow, piloselle, the extracts of ash, the extracts of cangzhu, the extracts of chrysanthellum indicum, of plants of the genus Armeniacea, Atractylodis platicodon, Sinnomenum, pharbitidis, Flemingia, of Coleus as C. forskohlii, C. blum, C. esquiro. scutellaroides, C. xanthantus and C. barbatus, as an extract of roots of Coleus barbatus, extracts of Ballote, Guioa, Davallia, Terminalia, Barringtonia, Tréma, Antirobia, Cecropia, Argania, Dioscoreae as Dioscorea opposita or Mexican extract, of Ammi vivisnagain, Siegesbeckia, in particular Siegesbeckia orientalis, plant extracts of the family Ericaceae, in particular extracts of blueberries (Vaccinium angustifollium) of Arctostaphylos uva ursi, Aloe vera, of plants containing sterols (especially of phytostaphyls), of plants of the genus Mantéresthal Rubia, in particular Rubia cordifolia^, of Guggal (extract from plants of the genus Commiphora, in particular Commiphora mukul),an extract of kola, chamomile, purple clover, Piper methysticum (Kava Kava de Sederma), Bacopa monieri (Bacocalmine™, Sederma) and sea buckthorn, Glycyrrhiza glabra, mulberry, melaleuca (tea tree), Larrea divaricata, Euscene de Rabeugle, gracilis, of Fibraurea recisa hirudinea, of Chaparral sorghum, of fleur de sunflower, of Enantia chlorantha, of Mitracarpe of the genus Spermacocea, of Buchu barosma, of Lawsonia inermis L., of Adiantium capillus-veneris L., of Chelidonium majus, of Luffa Marican, of Japanese cylinder, Marican. (Citrus reticulata blanco var. unshiu), of Camelia sinensis, of Imperata cylindrical, of Glaucium flavum, of Cupressus sempervirens, of Polygonatum multiflorum, of Loveyly hemsleya, of Sambucus nigra, of Phaseolus lunatus, of Centaurium, of Macrocystis pyriferastis, of Tumerarahena, of Tumerarus asphodeloides, Portulaca pilosa, Humulus lupulus, Arabica coffee, Ilex paraguariensis, Globularia cordifolia, Oxydendron arboreum,Albizzia julibrissin, Zingiber zerumbet smith, As-tragalus membranaceus, Atractylodes macrocephalae, Plantago lanceolata, Leontopodium alpinum (or eldelweiss), Mirabilis jalapa, Apium graveolens, Marrubium vulgare, Buddleja davidii Franch., Monarda didyma, Lavandula an-gustifolia or orchids. ,

[0199] The compositions according to the present invention may comprise peptides, including, but not limited to, di-, tri-, tetra-, penta- and hexapeptides and their derivatives. According to a particular embodiment, the concentration of the additional peptide, in the composition, varies between 1x10 7% and 20%, preferably between 1x10 6% and 10%, preferentially between 1x10 5% and 5%, by weight.

[0200] In the context of the present invention, the term "peptide" herein refers to peptides containing 10 or fewer amino acids, their derivatives, isomers and complexes with other species such as a metal ion (eg copper, zinc, manganese, magnesium, and others). The term "peptides" refers to both natural peptides and (bio)synthetic peptides. It also refers to compositions containing peptides that occur in nature and / or are commercially available.

[0201] Non-limiting examples of dipeptides that can be used in the context of the present invention include Carnosine (beta-AH), YR, VW, NF, DF, KT, KC, CK, KP, KK, TT, PA, PM or PP.

[0202] Non-limiting examples of tripeptides include RKR, HGG, GHK, GGH, GHG, GKH, KPK, KFK, KavaK, K[3AK, KabuK, KacaK, KPK, KMOK, KM02K, PPL, PPR, SPR, QPA, LPA or SPA.

[0203] Non-limiting examples of tetrapeptides are KTFK (SEQ ID NO: 7), GQPR (SEQ ID NO: 8), RSRK (SEQ ID NO: 9), KTAK (SEQ ID NO: 10), KAYK (SEQ ID NO: 11), KFYK (SEQ ID NO: 12), TKPR (SEQ ID NO: 13), AVPG (SEQ ID NO: 14), VPGA (SEQ ID NO: 15), LKLE (SEQ ID NO: 16), ELED (SEQ ID NO: 17) or LLAN (SEQ ID NO: 18).

[0204] Non-limiting examples of pentapeptide are KTTKS (SEQ ID NO: 19) and KTSKS (SEQ ID NO: 20).

[0205] Non-limiting examples of hexapeptides are GKTTKS (SEQ ID NO: 21) or VGVAPG (SEQ ID NO: 22).

[0206] Other peptides that can be used in the context of the present invention can be chosen from, without this list being limiting: lipophilic derivatives of peptides, preferably palmitoyl and myristoyl derivatives, and complexes with the metal ions mentioned above (eg copper complex of the tripeptide HGG).

[0207] Preferred dipeptides include, for example, N-Palmitoyl-[3-Ala-His, Pal-KT, Pal-RT (Sederma). Preferred tripeptides include, in particular, the copper derivative of HGG (Lamin™, Sigma), Pal-GHK, lipospondin (N-Elaidoyl-KFK) and its conservatively substituted analogues, N-Acetyl-RKR-NH2 (Peptide CK+), Pal-KavaK, Pal-K[3AlaK, Pal-KabuK, Pal-KacaK, Pal-KMO2K, N-Biot-GHK (Sederma) and their derivatives.

[0208] Mention may also be made here of the anti-aging tripeptides of general formula X-Pro*-Pro*-Xaa-Y described in application WO2015 / 181688 with Xaa chosen from Leu, Arg, Lys, Ala, Ser, and Asp, at the N-terminal end, X chosen from H, -CO-Ri and -SO2-Ri and at the C-terminal end Y chosen from OH, ORb NH2, NHRi or NRiR2, Ri and R2 being, independently of one another, chosen from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and / or sulfur-containing, said group being able to have in its skeleton a heteroatom in particular O, S and / or N, and Pro* corresponding to Proline, an analog or a derivative of this one; including for example Myr-PPL-OH and Myr-PPR-OH.

[0209] Mention may also be made here of the pro-pigmenting and / or pro-MEC dipeptides and tripeptides of general formula X-(Xaal)n-Pro*-Xaa2-Y described in application WO2014 / 080376, with n=0, 1 or 2, Xaal a hydrophobic amino acid chosen from Ala, Val, Met, Leu, Iso, Phe, Pro, and analogues or derivatives thereof; or a polar amino acid chosen from Ser, Thr, Tyr, Asp, Glu and derivatives and analogues thereof; and when n=2 the two Xaal amino acids may be identical or different; Xaa2 a hydrophobic amino acid chosen from Ala, Val, Met, Leu, Iso, Phe, and analogues or derivatives thereof; a basic amino acid chosen from Arg, Lys, His, and derivatives and analogues thereof; at the N-terminus of the peptide, X is chosen from H, -CO-Ri and -SO2-Ri;at the C-terminal end of the peptide, Y is chosen from OH, ORi, NH2, NHRi or NRiR2, Ri and R2 being, independently of one another, chosen from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and / or sulfur-containing, said group being able to have in its skeleton a heteroatom in particular O, S and / or N; Pro* corresponding to Proline, an analog or a derivative thereof; including for example the peptides Pal-SPR-OH, Pal-PPR-OH, Pal-QPA-OH, Pal-LPA-OH, Myr-SPA-OH, Pal-PM-OH, Pal-PA-OH and Pal-PP-OH. ;

[0210] Tetrapeptide derivatives that may be used in the present invention include, but are not limited to, Ela-KTAK (SEQ ID NO: 23), Ela-KAYK (SEQ ID NO: 24), Ela-KFYK (SEQ ID NO: 25), Pal-GQPR (SEQ ID NO: 3) or Pal-KTFK (SEQ ID NO: 5).

[0211] Useful pentapeptide derivatives include, but are not limited to, Pal-KTTKS (SEQ ID NO: 2), Pal-KTSKS (SEQ ID NO: 4), Pal-YGGFXaa (SEQ ID NO: 6) with Xaa being Leu or Pro.

[0212] Useful hexapeptide derivatives include, but are not limited to, Pal-HLDIIXaa (SEQ ID NO: 26) with Xaa being Trp, Phe, Tyr, Tic, 7-hydroxy-Tic or Tpi, or a mixture thereof, Pal-GKTTKS (SEQ ID NO: 27), Pal-VGVAPG (SEQ ID NO: 1).

[0213] Preferred commercially available compositions offered by Sederma:

[0214] - tripeptides or derivatives include in particular Biopeptide-CL™, Maxilip™, or Procapil™ including GHK;

[0215] - tetrapeptides or derivatives include: RIGIN™, Eyeliss™, which contain Pal- GQPR (SEQ ID NO: 3) and an excipient;

[0216] - pentapeptides or derivative such as Matrixyl ™ source of Pal-KTTKS (SEQ ID NO: 2).

[0217] We can also cite:

[0218] - the mixture Pal-GHK and Pal-GQPR (SEQ ID NO: 3) (Matrixyl™ 3000), and

[0219] - the mixture Pal-GHK and Pal-VGVAPG (SEQ ID NO: 1) (Biobustyl™).

[0220] The following commercial peptides may also be mentioned as additional active ingredients:

[0221] - Vialox™, Syn-ake™ (|3-Ala-Pro-Dab-NH-Bzl) or Syn-Coll™ (Pal-Lys-Val-Lys-OH) sold by the company Pentapharm,

[0222] - Argireline™ (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH2 (SEQ ID NO: 28) (INCI Name = Acetyl hexapeptide-3), Leuphasyl™ (Tyr-D-Ala-Gly-Phe-Leu (SEQ ID NO: 29)), Aldenine™ (Gly-His-Lys), Trylagen™ (INCI Name = Pseudoalteromonas Ferment Extract, Hydrolyzed Wheat Protein, Hydrolyzed Soy Protein, Tripeptide-10 Citrulline (reaction product of citrulline and Tripeptide-10 (synthetic peptide consisting of aspartic acid, isoleucine and lysine)), Tripeptide-1), Eyeseryl™ (Ac-[3-Ala-His-Ser-His (SEQ ID NO: 30)), Serilesine™ (Ser-Ile-Lys-Val-Ala-Val (SEQ ID NO: 31)) or Decorinyl™ (INCI Name: Tripeptide-10 Citrulline = product of the reaction of Citrulline and Tripeptide-10 (synthetic peptide consisting of aspartic acid, isoleucine and lysine) sold by the company Lipotec,

[0223] - Collaxyl™ (Gly-Pro-Gln-Gly-Pro-Gln (SEQ ID NO: 32)) or Quintescine™ (Cys-Gly) sold by the company Vincience,

[0224] - Cytokinol™LS (casein hydrolyzate) sold by Laboratoires Serobio- logic / cognitive,

[0225] - Kollaren™ (Gly-His-Lys), 1TP2000™ (Pal-Val-Tyr-Val) or Meliprene™ sold by the European Institute of Cell Biology,

[0226] - Neutrazen™ (Pal-His-D-Phe-Arg-NH2) sold by the company Innovations, or

[0227] - BONT-L-Peptide™, Timp-Peptide™ or ECM Moduline™ (INCI Name = Palmitoyl Tripeptide-28: product of the reaction of palmitic acid and Tripeptide-28 (synthetic peptide consisting of arginine, lysine and phenylalanine) sold by the company Infinitec Activos.

[0228] It is also possible to combine the invention with one or more cyclic peptides, in particular those extracted from linseed oil described in the Applicant's patent application FRI850845. Serum formula

[0229] [Tables 1] Raw materials % INCI name Phase A Water qs Aqua / water Sodium benzoate qs Sodium benzoate Potassium sorbate qs Potassium sorbate Phase B Butylene glycol 5.50 Butylene Glycol Xanthan gum 0.25 Xanthan Gum Arlasolve DMI PC 2.50 Dimethyl Isosorbide Phase C Water 0.40 Aqua / water Lactic acid 0.04 Lactic acid Phase D Active ingredient according to the invention 3.0 Operating mode:

[0230] Weigh and homogenize A. Weigh and homogenize B. Add B to A while stirring, homogenize. Weigh and homogenize C. Add C to A+B, homogenize. Add D to A+B+C, homogenize. Milky Mist Formula

[0231] [Tables2] Raw materials % INCI name Phase A Water qs Aqua / water Potassium sorbate 0.10 Potassium sorbate Phase B Glycerin 5.00 Glycerin Microcare® Emollient DCP 1.50 Pentylene Glycol (and) Caprylyl Glycol (and) Decylene Glycol Amigum 0.20 Sclerotium Gum Phase C SP Span™ 20 MB AL 1.00 Sorbitan Laurate Phase D Active ingredient according to the invention 3.0 Operating mode:

[0232] Weigh and homogenize A. Weigh and homogenize B. Add B to A while stirring. Activate A+B by mixing them vigorously. Weigh and homogenize C. Add C to A+B, homogenize. Add D to A+B+C, homogenize. 5. Results of in vitro tests

[0233] The mixture of oligosaccharides obtained in point 3. was used for the in vitro tests carried out below, noted “active ingredient according to the invention”.

[0234] 5.1. Protection against loss of radiance of the complexion and the appearance of signs of v skin aging 5.1.1 Fight against oxidation

[0235] 5.1.1.1 Effect on reactive oxygen species Protocol

[0236] Human keratinocytes (KH) are cultured to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 28 hours and then receive a fluorescent probe intended to mark the Intracellular production of ROS. After incorporation, cells become sensitive to oxidants. Then, the cells receive or not an agent intended to create ROS (oxidative stress). A fluorescence reading (e.g.: 490 nm / em: 520 nm) allows the quantity of intracellular ROS to be estimated. The number of cells is estimated using the Hoechst method (DNA staining) to weight the data obtained. Results

[0237] Variation in ERO production in KH. Effect of 2% of the active ingredient according to the invention compared to the control (n=6):

[0238] [Tables3] Variation (%); significance Control case Reference 2% of the active ingredient according to the invention - 39.6%; p<0.01

[0239] The results show that 2% of the active ingredient according to the invention significantly reduces the production of EROs by -39.6% (p<0.01). 5.1.1.2. Effect on Nrf2 protein Protocol

[0240] KH cells are cultured to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 2 hours or 24 hours. Then, immunocytochemical labeling of the Nrf2 protein and its passage from the cytoplasm to the nucleus was followed by image analysis. Results

[0241] Variation of Nrf2 translocation as a function of time in a KH culture. Effect of 3% of the active ingredient according to the invention compared to the control case (n=4):

[0242] [Tables 4] Control case: Contact with 0.1% DMSO Contact with 3% of the active ingredient according to the invention Variation (%); significance After 2 hours of contact Reference 1 +41%; p<0.01 After 24 hours of contact Reference 2 +36%; p<0.05

[0243] The results show that 3% of the active ingredient according to the invention significantly increases the translocation of the Nrf2 protein by +41% (p<0.01) from 2 hours of contact and +36% (p<0.05) after 24 hours of contact. 5.1.1.3. NQO-1 Protocol

[0244] KH cells are grown to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 6 hours. Then the KH mats are rinsed and the cells are ground to extract their mRNA. These mRNAs are then converted into small DNA sequences which are analyzed after deposition on DNA chips and amplification by a method similar to QRT-PCR. The variations in mRNA due to the ingredient according to the invention are compared to the control case. Results

[0245] The results show that 3% of the active ingredient according to the invention significantly increases the production of the NQO-1 protein (+67%, p<0.01). 5.1.1.4. HMOX-1 Protocol

[0246] Firstly, KHs are grown to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 54 hours. The cells are rinsed, dried and stored frozen. The quantity of HMOX-1 is measured by ELIS A method, after extraction.

[0247] The amount of total protein was evaluated after extraction, by the bicinchonic acid method to weight the data obtained.

[0248] In a second step, the cells are placed in contact with 500 pM of Trolox instead of the ingredient according to the invention. Trolox is a close and hydrophilic analogue of vitamin E known for its antioxidant activity used to limit damage linked to oxidative stress. Results

[0249] Variation in the stimulation of HMOX-1 production. Effect of 3% of the active ingredient according to the invention at 54 hours compared to the control case (n=4):

[0250] [Tables5] Variation (%); significance Control case 1 Reference 1 3% of the active ingredient according to the invention + 65%; p<0.01 Control case 2 Reference 2 500 pM of Trolox + 24%; p<0.01

[0251] The results show that the stimulation of HMOX-1 production is significantly increased in both cases, but that 3% of the active ingredient according to the invention stimulates more HM0X1 production (+65%; p<0.01 vs its control) compared to 500 pm of Trolox (+24%; p<0.01 vs its control). 5.1.2. Combating cellular hypoxia

[0252] 5.1.2.1. Mitochondrial potential AT, promoting force Protocol

[0253] KHs are cultured to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 28 hours and then they are labeled with a fluorescent probe.

[0254] A reading of these probes by fluorescence makes it possible to measure the two forms and to establish the monomer / polymer ratio (520 nm / 590 nm). A high value of this ratio is less favorable to the survival of the cell than a lower value.

[0255] The number of cells is estimated using the Hoechst method (DNA staining) to weight the obtained data. Results

[0256] Variation of mitochondrial potential AT in KH. Effect of 3% of the active ingredient according to the invention compared to the control case (n=6):

[0257] [Tableauxô] Variation (%); significance Control case Reference 3% of the active ingredient according to the invention + 33.2%; p<0.01

[0258] The results show that 3% of the active ingredient according to the invention significantly increases the mitochondrial AT potential by +33.2% (p<0.01).

[0259] 5.1.2.2. Protection of respiration, oxygen consumption Protocol

[0260] KHs are cultured to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 24 hours. The KHs are then detached from their support and deposited in equivalent quantities in the wells of a plate having sensors which allow the measurement of oxygen consumption by these cells as a function of time. Results

[0261] Variation in oxygen consumption (oxygen pressure per minute: pO2) in KH. Effect of 3% of the active ingredient according to the invention compared to the control case (n=6):

[0262] [Tables?] Variation (%); significance Control case Reference 3% of the active ingredient according to the invention + 23.6%; p<0.05

[0263] The results show that 3% of the active ingredient according to the invention has a beneficial effect on cell respiration by stimulating it by +23.6% (p<0.05). 5.1.2.3. ATP synthesis Protocol

[0264] KH cells are grown to confluence in their culture medium. The cells are then placed in contact with the ingredient according to the invention for 26 hours. Intracellular ATP is extracted and measured by bioluminescence, the signal obtained being proportional to the quantity of ATP in the cells.

[0265] The number of cells is estimated using a propidium iodide fluorescent probe to weight the data obtained. Results

[0266] Variation of ATP synthesis in KH. Effect of 3% of the active ingredient according to the invention compared to the control case (n=6):

[0267] [Tables8] Variation (%); significance Control case Reference 3% of the active ingredient according to the invention + 84%; p<0.01

[0268] The results show that 3% of the active ingredient according to the invention significantly increases ATP synthesis by +84% (p<0.01) in a dose-dependent manner compared to the control case. 5.1.3. Strengthening the barrier function 5.1.3.1. Thickness of the stratum corneum Protocol

[0269] Firstly, equivalent skins are prepared and placed in contact with the ingredient according to the invention for 2 days. The skins are then marked with Hematoxylin and Eosin to visualize the macrostructure of the different layers of the skin. This marking is followed by image analysis. Results

[0270] Variation in the thickness of the stratum corneum. Effect of 3% of the active ingredient according to the invention compared to the control case (n=6):

[0271] [Tables9] Variation (%); significance Control case Reference 3% of the active ingredient according to the invention + 43.4%; p<0.01

[0272] The results show that 3% of the active ingredient according to the invention significantly increases the thickness of the stratum corneum by + 43.4% (p<0.01). 5.1.3.2. Involucrin and Corneodesmosine Protocol

[0273] Equivalent skins are prepared and placed in contact with the ingredient according to the invention for 2 days. The skins are then marked using antibodies directed against involucrin and corneodesmosine. This marking is followed by image analysis (N = 57 to 60 photos per case).

[0274] The number of cells is estimated using the Hoechst method (DNA staining) to weight the obtained data. Results

[0275] Variation in the synthesis of corneodesmosine and involucrin. Effect of 3% of the active ingredient according to the invention compared to the control case (n=5):

[0276] [TableauxlO] Involucrin Variation (%); significance Corneodesmosine Variation (%); significance Control case Reference 1 Reference 2 3% of the active ingredient according to the invention + 42.0%; p<0.01 + 46.0%; p<0.01

[0277] The results show that 3% of the active ingredient according to the invention significantly increases the production of involucrin (+ 42%; p<0.01) and corneodesmosine (+ 46%; p<0.01). The quality of the barrier function is therefore improved. 5.1.4. Strengthening the hydration function Protocol

[0278] Skin explants are prepared and the ingredient according to the invention is deposited on their surfaces for 3 days. Then, the explants are sectioned into 7 μm thin sections and labeled using antibodies directed against hydration markers: filaggrin and aquaporin-3. This labeling is followed by image analysis (N = 41 to 45 photos per case). Results

[0279] Variation in the synthesis of filaggrin and aquaporin-3 in skin explants. Effect of 3% of the active ingredient according to the invention compared to the control case (n=3):

[0280] [Tableauxll] Variation (%); significance Aquaporin-3 + 11.0%; p<0.01 Filaggrin + 10.0%; p<0.01

[0281] The results show that 3% of the active ingredient according to the invention significantly increases the two markers of hydration.

[0282] 5.1.5. Strengthening the function of the dermo-epidermal junction (DEJ) Protocol

[0283] The same protocol as for filaggrin and aquaporin-3 was used for claudin-1, collagen IV and collagen VIL. Labeling is followed by image analysis (N= 41 to 45 photos per case). Results

[0284] Variation in the synthesis of Claudin-1, collagen IV and collagen VII in skin explants. Effect of 3% of the active ingredient according to the invention compared to the control case (n=3):

[0285] [Tablesl2] Variation (%); significance Collagen IV + 26.0%; p<0.01 Collagen VII + 27.0%; p<0.01 Claudin-1 + 7.0%; p<0.01

[0286] The results show that 3% of the active ingredient according to the invention significantly increases the markers of the JDE (collagen IV and collagen VII) and the marker of tight junctions (claudin-1). 5.1.6. Strengthening microbiota function Protocol

[0287] The ingredient according to the invention is brought into contact with a culture of Staphylococcus epidermidis (ATCC 14990) at 4.105CFU* / mL, all at 37°C in a culture medium allowing the growth of this bacterium. The growth kinetics are monitored by optical density (OD) measurements at 600 nm. Results

[0288] Growth kinetics of S. epidermidis over time. Effect of 3% of the active ingredient according to the invention compared to the control case (n=3):

[0289] [Tables 13] Variation between T=0 and 12 hours (%); significance Variation between T=0 and 16 hours (%); significance Control case Reference 1 Reference 2 3% of the active ingredient according to the invention + 1120%; p<0.01 + 1497%; p<0.01

[0290] The results show that 3% of the active ingredient according to the invention significantly increases the growth of Staphylococcus epidermidis. 5.2. Protection against pollution

[0291] The tests were carried out in the presence of atmospheric MPUs to simulate the effect of pollution. 5.2.1. Fight against oxidation

[0292] 5.2.1.1. Action on reactive oxygen species Protocol

[0293] The effects of atmospheric pollutants on the formation of ROS were evaluated. The same protocol as in point 5.1.1.1. is used and an MPU solution is brought into contact with the cells cultured and labeled as before. Results

[0294] Variation of ERO production in KH (n=6) with effect of atmospheric pollutants. Effect of 2% of the active ingredient according to the invention compared to the control with MPU (n=6):

[0295] [Tablesl4] Variation (%); significance Control case without MPU Reference 1 Control case with MPU + 26.9%; p<0.01 Reference 2 2% of the active ingredient according to the invention, with MPU - 41.9%; p<0.01

[0296] The results show that the polluting microparticles increase the production of EROs by + 26.9% (p<0.01) and that 2% of the active ingredient according to the invention makes it possible to counteract the effects of pollution by significantly reducing the production of oxidative radical species by - 41.9% (p<0.01). 5.2.1.2. Nrf2 protein Protocol

[0297] The effects of air pollutants on the synthesis of Nrf2 protein have been evaluated. The same protocol as in point 5.1.1.2. is used and a solution of MPU or Benzo[a]pyrene is brought into contact with the cultured and labeled cells as before. Results

[0298] Variation of Nrf2 translocation as a function of time in a KH culture. Effect of 3% of the active ingredient according to the invention, atmospheric MPU solution and Benzo[a]pyrene (B[a]P):

[0299] [Tables 15] Control: Contact with 0.1% DMSO Contact with MPU; Contact with B[a]P; Contact with 3% of the active ingredient according to the invention Variation (%); significance Variation (%); significance Variation (%); significance After 2 hours of contact Reference 1 + 136%; p<0.01 + 58%; p<0.05 + 41%; p<0.01 After 24 hours of contact Reference 2 + 401%; p<0.01 + 199%; p<0.01 + 36%; p<0.05

[0300] The results show that the translocation of the Nrf2 protein from the cytoplasm to the nucleus is significantly increased in all three cases, but that there are large differences in both duration and intensity.

[0301] Concerning atmospheric MPUs, the results show a strong increase after 2 hours of contact (+ 136%; p<0.01 compared to the control case) and a very strong increase in translocation after 24 hours of contact (+401%; p<0.01).

[0302] Concerning Benzo[a]pyrene, one of the best known atmospheric pollutants which is carried by MPU, the results show a more moderate increase in translocation of + 58% (p<0.05) after 2 hours of contact and an exponential increase after 24 hours of contact (+ 199%; p<0.01).

[0303] Concerning 3% of the active ingredient according to the invention, the results show an increase in the translocation of the Nrf2 protein of + 41% (p<0.01) after 2 hours of contact and of + 36% (p<0.05) after 24 hours of contact. These results remain moderate compared to the other two cases (MPU and B[a]P), but significant compared to the control case. This underlines the controlled “start-up” of the cell detoxification system in order to combat pollutants. 5.2.2. Combating cellular hypoxia

[0304] 5.2.2.1. Mitochondrial potential A*P, promoting force Protocol

[0305] The effects of atmospheric pollutants on the mitochondrial potential AW were evaluated. The same protocol as in point 5.1.2.1 is used and a solution of atmospheric MPU is brought into contact with the cells cultured and labeled as before. Results

[0306] Variation of mitochondrial potential AW in KH. Effect of 3% of the active ingredient according to the invention compared to the control case with or without MPU (n=6):

[0307] [Tables 16] Variation (%); significance Control case without MPU Reference 1 Control case with MPU - 72.0%; p<0.01 Reference 2 3% of the active ingredient according to the invention, with MPU + 49.6%; p<0.01

[0308] The results show that the polluting microparticles reduce the mitochondrial AW potential by -72% (p<0.01) and that 3% of the active ingredient according to the invention makes it possible to counteract the effects of pollution by significantly increasing the mitochondrial AW potential by +49.6% (p<0.01).

[0309] 5.2.2.2. Protection of respiration, oxygen consumption Protocol

[0310] The effects of atmospheric pollutants on oxygen consumption by KH were evaluated. The same protocol as in point 5.1.2.2. is used and a solution of atmospheric MPU is brought into contact with the cells cultured as before. Results

[0311] Variation in oxygen consumption (oxygen pressure per minute: pO2) in KH. Effect of 3% of the active ingredient according to the invention compared to the control case with or without MPU (n=6):

[0312] [Tablesl7] Variation (%); significance Control case without MPU Reference 1 Control case with MPU - 29.8%; p<0.01 Reference 2 3% of the active ingredient according to the invention, with MPU + 24.2%; p<0.01

[0313] The results show that the polluting microparticles reduce oxygen consumption by cells by -29.8% (p<0.01) and that 3% of the active ingredient according to the invention makes it possible to counteract the effects of pollution by significantly increasing cellular respiration by +24.2% (p<0.01). 5.2.2.3. Protection of ATP production Protocol

[0314] The effects of atmospheric pollutants on ATP synthesis by KH were evaluated. The same protocol as in point 5.1.2.3. was used and a solution of atmospheric MPU was brought into contact with the cells cultured as before. Results

[0315] Variation of ATP synthesis in KH. Effect of 3% of the oligosaccharide mixture compared to the control case with or without MPU (n=6):

[0316] [Tables 18] Variation (%); significance Control case without MPU Reference 1 Control case with MPU - 21%; p<0.01 Reference 2 3% of the active ingredient according to the invention, with MPU + 74%; p<0.01

[0317] The results show that the polluting microparticles reduce ATP production by -21% (p<0.01) and that 3% of the active ingredient according to the invention makes it possible to counteract the effects of pollution by significantly increasing ATP synthesis by +74% (p<0.01). 5.2.2.4. Conclusion

[0318] All of the in vitro results presented above show that the mixture of oligosaccharides according to the invention protects cells against oxidation and hypoxia with the aim of detoxifying cells from the effects of micropollutants. 5.2.3. Strengthen the skin barrier 5.2.3.1. Thickness of the stratum corneum Protocol

[0319] The effects of atmospheric pollutants on the thickness of the stratum corneum are evaluated. The same protocol as in point 5.1.3.1. is used and a solution of atmospheric MPU is brought into contact with equivalent skins. Results

[0320] Variation in the thickness of the stratum corneum. Effect of 3% of the active ingredient according to the invention (n=4):

[0321] [Tablesl9] Variation (%); significance Control case without MPU Reference 1 Control case with MPU - 61.3%; p<0.01 Reference 2 3% of the active ingredient according to the invention, with MPU + 136.4%; p<0.01

[0322] The results show that the polluting microparticles reduce the thickness of the stratum corneum by -61.3% (p<0.01) and that 3% of the active ingredient according to the invention makes it possible to counteract the effects of pollution by significantly increasing the thickness of the stratum corneum by +136.4% (p<0.01). This demonstrates the positive effect of the ingredient according to the invention against the deleterious effects of pollution.

[0323] 5.3. Comparative results

[0324] The monosaccharides and monosaccharide acids of the oligosaccharide mixture, namely N-acetyl-quinovosamine, galacturonic acid, glucuronic acid and rhamnose were tested alone and in mixture to allow comparison with the active ingredient according to the invention. The quantities tested are:

[0325] - 50 ppm of rhamnose;

[0326] - 50 ppm of N-acetyl-quinovosamine;

[0327] - 50 ppm of galacturonic acid;

[0328] - 50 ppm of glucuronic acid;

[0329] - In the mixture of the 3 saccharides: 50 ppm of N-acetyl-quinovosamine, 50 ppm galacturonic acid and 50 ppm glucuronic acid;

[0330] - In the mixture of 4 saccharides: 50 ppm of N-acetyl-quinovosamine, 50 ppm rhamnose, 50 ppm galacturonic acid and 50 ppm glucuronic acid. 5.3.1. ATP synthesis

[0331] Protocol

[0332] The same protocol as in point 5.1.2.3. was used.

[0333] Results

[0334] Variation of ATP synthesis in KH. Comparison of the effects of saccharides alone or in mixture and of 3% of the mixture of oligosaccharides according to the invention compared to the control case (n=8):

[0335] [Tables20] Effect on ATP production Control case Reference 50 ppm rhamnose 0 50 ppm glucuronic acid 0 50 ppm galacturonic acid 0 50 ppm N-acetyl-quinovosamine 0 Mixture of the 3 saccharides 0 3% of the active ingredient according to the invention ++

[0336] The results show that the monosaccharides alone or in combination have no effect on the production of ATP, unlike the active ingredient according to the invention. It is therefore the oligosaccharides according to the invention, i.e. the monosaccharides linked to each other, which act on the production of ATP. 5.3.2. Kinetics of S. epidermidis

[0337] Protocol

[0338] The same protocol as in point 5.1.6. was used. Results

[0339] Growth kinetics of S. epidermidis over time. Comparison of the effects of saccharides alone or in mixture and of 3% of the oligosaccharide mixture compared to the control case (n= 7):

[0340] [Tables21] Effect on the production of Staphylococcus epidermidis Control case Reference 50 ppm rhamnose 0 50 ppm glucuronic acid 0 50 ppm galacturonic acid 0 50 ppm N-acetyl-quinovosamine 0 Mixture of the 4 saccharides 0 3% of the active ingredient according to the invention ++

[0341] The results show that the monosaccharides alone or in combination have no effect on the production of Staphylococcus epidermidis, unlike the active ingredient according to the invention. It is therefore the oligosaccharides according to the invention, i.e. the monosaccharides linked to each other, which act on the growth of S. epidermidis. 6. Results of in vivo tests Product tested

[0342] The serum described in Table 1 above vs. a placebo serum, i.e. with the same chassis as the serum without the active ingredient according to the invention. Principle

[0343] The evaluation of the effectiveness of the cosmetic treatment according to the invention was carried out on a total of 181 volunteers during two complementary studies: one on a Caucasian panel and one on a Chinese panel. The panelists had to have a dull complexion and an uneven skin texture.

[0344] For the evaluation of metrological means (measurements), expert analyses (judgments) and self-assessments (perceived effects) were used. Protocol

[0345] The study on Caucasian volunteers was conducted in France on a maximum of 54 female volunteers with an average age of 38 years (20 - 50 years).

[0346] The study on Chinese volunteers was conducted in China on a maximum of 127 volunteers, men and women, with an average age of 35 years (18 - 50 years). Type of study, duration, applications

[0347] The volunteers were not aware of the type of product (active according to the invention or placebo).

[0348] The panel of 54 Caucasian women was divided into two and different measurements and entries were made before and after the applications, according to Table 22 below:

[0349] [Tables22] T0 (D0) T = 28 days (D28) T = 56 days (D56) Radiance assessment Photos Photos / Microrelief assessment Fringe projection Fringe projection / Hydration assessment MoistureMeter-D TM MoistureMeter-D™ MoistureMeter-D™ Firmness assessment Cutometer® MPA580 / Cutometer® MPA580 Self-assessment / / Questionnaire

[0350] The panel of 127 Chinese volunteers applied four fabric masks impregnated with the serum according to the invention in 14 days for 15 minutes. The perception of the effect of the serum according to the invention in the mask was evaluated using a self-assessment questionnaire after the first and after the fourth and final application. Statistics

[0351] For instrumental parameters, statistical calculation is performed using a Student t-test or, if necessary, a non-parametric Wilcoxon or Mann-Whitney test. One-sided tests were performed on unpaired series.

[0352] For the questionnaire values, the statistical calculation is performed using a Chi2 test. The level of 0.05 is chosen as the minimum acceptance point for statistical significance. 6.1. Assessment of complexion radiance

[0353] The skin radiance index (SRI “Skin radiance Index”) results from a calculation which takes into account four areas of the face which are the forehead, the nose, the dark circles and the cheeks, and three colorimetric components which are L* which represents the brightness or clarity, H which represents the hue of a color and IWA (“Individual White Angle”) which is an index developed by the company Newtone Technologie to evaluate the whiteness of the skin by taking into account the parameters L*, a* and b* (CIELAB or CIELCH color space). Protocol

[0354] A Headscan V05™ camera bench (Orion Concept, France) is used to acquire photographs of the face in cross-polarized mode in order to remove natural shine.

[0355] For this evaluation, only 52 Caucasian volunteers participated: 26 applied the serum containing the active ingredient according to the invention and 26 applied the placebo serum. The results below are an average of the measurements taken on the different parts of the face. Results

[0356] Variation in skin radiance index after 28 days of application. Effect of 3% of the active ingredient according to the invention compared to the placebo (N=52):

[0357] [Tables23] Serum according to the invention Placebo serum Variation between D0 and D28 (%); significance +3.1%; p<0.05 -0.2%; dns* % improvement of the serum according to the invention vs. the placebo serum; significance vs. Placebo +3.3%; p<0.05

[0358] * dns: non-significant data

[0359] The results show that the skin radiance index is significantly increased with the serum according to the invention (+3.3%; p<0.05 vs. placebo). This shows that the ingredient according to the invention improves skin radiance. 6.2. Evaluation of the microrelief

[0360] Several parameters are studied to demonstrate the interest of the active ingredient according to the invention on the cutaneous microrelief.

[0361] The first parameter is called complexity. It represents the ratio between the real surface area and the ideal apparent surface area.

[0362] The second parameter is called maximum amplitude. It corresponds to the average height of the highest reliefs. It reflects skin alterations.

[0363] The third parameter is called roughness. It corresponds to the average roughness of the facial skin. The greater it is, the less smooth the skin is. Protocol

[0364] The projection of shadows of features (fringes) onto the face was carried out and their deformations, linked to the relief of the skin, measured by triangulation. This technology allows a three-dimensional reconstruction of the relief.

[0365] For this evaluation, only 53 Caucasian volunteers were included: 26 applied the serum containing the active ingredient according to the invention and 27 applied the placebo serum.

[0366] The measurements were taken on the cheek and a surface roughness analysis from the captures was carried out to qualify the more or less smooth appearance of the skin texture (microrelief, pores, micro-fine lines). The results below are an average of the different measurements taken. Results

[0367] Variation in complexity, maximum amplitude and roughness after 28 days of application. Effect of 3% of the active ingredient according to the invention compared to placebo (N=53):

[0368] [Tableaux24] % variation between D0 and D28; significance vs. D0 % improvement of the serum according to the invention vs. the placebo serum; significance vs. placebo Complexity Serum according to the invention -8%; p<0.01 -5.7%; p<0.05 Placebo serum -2.3%; dns Maximum amplitude Serum according to the invention -4.9%; p<0.01 -4.1%; p<0.05 Placebo serum -0.8%; dns Roughness Serum according to the invention -3.8%; p<0.01 -3.2%; p<0.05 Placebo serum -0.6%; dns

[0369] The results show that the serum according to the invention significantly reduces the complexity, maximum amplitude and roughness of the skin after 28 days of application. A reduction in these three parameters reflects smoothing and brighter, more radiant facial skin. 6.3. Hydration assessment

[0370] Skin hydration contributes to the formation and maintenance of an effective skin barrier against external aggressions, to fight against allergens, to limit water loss, to make it stronger, smoother, firmer and contributes to skin radiance. Protocol

[0371] The MoistureMeter-D™ is a device for measuring the quantity of skin water using the principle of impedancemetry.

[0372] During this study, 6 areas were studied in parallel: forehead, temples, cheekbones, cheeks, jowls, chin. The results below are an average of the different measurements taken and allow us to express the overall hydration of the volunteers' faces. The measurements were taken at least 12 hours after the last application.

[0373] For this evaluation, 54 Caucasian volunteers were included: 27 applied the serum containing the active ingredient according to the invention and 27 applied the placebo serum. Among the 54 volunteers, 24 had dry skin: 13 applied the serum containing the active ingredient according to the invention and 11 applied the placebo serum. Results

[0374] Variation in hydration after 28 days and 56 days of application. Effect of 3% of the active ingredient according to the invention compared to the placebo:

[0375] [Tables25] All skin types (N=54) Dry skin types (N=24) Serum according to the invention Placebo serum Serum according to the invention Placebo serum % variation between D28 and ; significance vs. D0 + 8.4%; p<0.01 0%; dns + 13.9%; p<0.01 + 4.5%; dns % improvement of the serum according to the invention vs. placebo serum; significance vs. placebo + 8.4%; p<0.01 + 9.4%; p<0.05 % variation between D56 and D0; significance vs. D0 + 15.0%; p<0.01 - 0.3%; dns + 21.4%; p<0.01 + 6.6%; p<0.05 % improvement of the serum according to the invention vs. placebo serum; significance vs. placebo + 15.3%; p<0.01 + 14.8%; p<0.01

[0376] The results show that the serum according to the invention significantly increases skin hydration after 28 days and 56 days of application compared to the placebo. Furthermore, the extraction of “dry skin” shows that the long-term moisturizing contribution of serum according to the invention also increases significantly.

[0377] The measurements having been carried out at least 12 hours after the last application of the product on the skin, the results show a real beneficial effect of skin hydration over the long term. 6.4. Evaluation of skin firmness

[0378] Skin firmness is one of the most sought-after parameters because it is linked to youth, skin resistance and tone. Protocol

[0379] The Cutometer® is used to measure the viscoelastic parameters of the skin. This device measures the deformation of a skin area, subjected to repeated mechanical suction stresses, as well as its recovery power. The device provides graphs of skin deformation as a function of time as shown in [Fig.l]. In this [Fig.l], the parameters Uf and RIO are indicated, representing respectively the extensibility and the "fatiguability" of the skin.

[0380] In this study, 3 measurements were taken on the right and left sides of the face. The results of extensibility are an average of the different measurements taken and the results of recovery power are an average of the last measurements taken on the right and left sides of the face.

[0381] For this evaluation, 54 Caucasian volunteers were included: 27 applied the serum containing the active ingredient according to the invention and 27 applied the placebo serum. Results

[0382] Variation in extensibility (Uf) and recovery power (RIO) after 56 days of application (N=54). Effect of 3% of the active ingredient according to the invention compared to the placebo:

[0383] [Tables26] Uf RIO Serum according to the invention Placebo serum Serum according to the invention Placebo serum % variation between D56 and D0; significance vs. D0 - 8.0%; p<0.01 - 2.3%; dns - 4.7%; p<0.05 - 0.2%; dns % improvement of the serum according to the invention vs. placebo serum; significance vs. placebo - 5.7%; p<0.05 - 4.5%; p<0.06

[0384] The results show that the serum according to the invention significantly reduces skin extensibility and fatigue, thus demonstrating an increase in skin firmness.

[0385] 6.5. Perception of the effects of the ingredient according to the invention 6.5.1. In the Caucasian panel Protocol

[0386] The volunteers' opinions were collected using a self-assessment questionnaire after 56 days of application of the serum containing the active ingredient according to the invention or the placebo serum. Results

[0387] Number of favorable opinions for each statement. Effect of 3% of the active ingredient according to the invention compared to the placebo:

[0388] [Tables27] % number of favorable opinions for the serum according to the invention; significance % number of favorable opinions for the placebo serum; significance % number of favorable opinions for the serum according to the invention vs. placebo serum; significance vs. placebo My skin is oxygenated and purified 67%; p<0.05 46%; dns + 11%; p<0.05 The product hydrates immediately and lastingly 83%; p<0.01 69%; p<0.005 + 14%; p<0.08 My skin looks more beautiful 89%; p<0.01 67%; p<0.005 + 22%; p<0.05

[0389] The results show that the serum according to the invention has significantly more favorable perceived effects compared to the placebo serum. The panelists find that their skin is more oxygenated, purified, immediately and lastingly hydrated, and looks more beautiful. 6.5.2. In the Chinese panel Protocol

[0390] The volunteers' opinions were collected using a self-assessment questionnaire after 56 days of application of a facial tissue mask impregnated with serum containing the active ingredient according to the invention or with placebo serum. Results

[0391] Number of favorable opinions for the serum according to the invention:

[0392] [Tables28] % of favorable opinions after the 1st application % of favorable opinions after the 4th application % of favorable opinions after the 1st application v5. the 4th application My complexion is less dull 78% 91% + 13% My complexion is clearer 82% 92% + 10% My skin is firmer 83% 89% + 6% My skin is softer 90% 94% + 4% My skin is more hydrated 90% 95% + 5% My skin is fresher 87% 93% + 6% The skin is revitalized / energized 87% 91% + 4% My skin is purified 87% 91% + 4% I look younger 77% 89% + 12% I look less tired 89% 96% + 7%

[0393] The results show that the perception of skin quality is improved after four close applications in time of a fabric mask impregnated with the serum according to the invention. 6.6. Conclusion

[0394] All of the in vivo results presented above show that the treatment according to the invention advantageously makes it possible to reinforce the radiant appearance of the skin, make it appear more luminous and invigorated, to hydrate it, smooth the relief, reinforce its firmness and reduce skin fatigue.

Claims

Claims

1. Exopolysaccharide characterized in that it comprises at least one monosaccharide unit which is N-acetyl-quinovosamine.

2. Exopolysaccharide according to claim 1, characterized in that it further comprises at least one monosaccharide acid unit chosen from glucuronic acid and galacturonic acid.

3. Exopolysaccharide according to claim 1 or 2, characterized in that it comprises at least one monosaccharide unit which is rhamnose.

4. Exopolysaccharide according to any one of claims 1 to 3, comprising between 15% and 55% of N-acetyl-quinovosamine expressed by weight relative to the total weight of the exopolysaccharide.

5. Exopolysaccharide according to any one of claims 1 to 4, comprising between 15% and 55% of glucuronic acid and / or between 15% and 55% of galacturonic acid, expressed by weight relative to the total weight of the exopolysaccharide.

6. Exopolysaccharide according to any one of claims 1 to 5, comprising between 15% and 55% of rhamnose expressed by weight relative to the total weight of the exopolysaccharide.

7. An exopolysaccharide according to any one of claims 1 to 6, composed of an equimolar mixture of N-acetyl-quinovosamine, glucuronic acid, galacturonic acid and rhamnose.

8. Exopolysaccharide according to any one of claims 1 to 7, characterized in that it comprises a chain of three units comprising N-acetyl-quinovosamine and two monosaccharide acid units chosen from glucuronic acid and galacturonic acid.

9. Exopolysaccharide according to one of the preceding claims, characterized by a molecular mass greater than or equal to 260,000 Da.

10. Exopolysaccharide according to any one of the preceding claims, characterized in that it is derived from the bacterial culture of the strain Pseudoalteromonas sp. deposited in the National Collection of Culture of Microorganisms (CNCM) of Paris under number 1-5893.

11. Oligosaccharide which can in particular be obtained by cleavage of the exopolysaccharide according to any one of the preceding claims, characterized in that it comprises N-acetyl-quinovosamine.

12. Oligosaccharide according to claim 11, characterized in that it has a degree of polymerization of between 2 and 12.

13. Oligosaccharide according to claim 11 or 12, characterized in that it further comprises at least two monosaccharide acid units chosen from glucuronic acid and galacturonic acid.

14. Oligosaccharide according to any one of claims 11 to 13, characterized in that it consists of N-acetyl-quinovosamine, glucuronic acid and galacturonic acid.

15. Oligosaccharide according to any one of claims 11 to 14, characterized in that it comprises the chain glucuronic acid, galacturonic acid and N-acetyl-quinovosamine.

16. A method for obtaining the exopolysaccharide according to any one of claims 1 to 10, comprising the following steps: - A first step of culturing a bacterial strain of the genus Pseudoalteromonas sp. deposited at the National Collection of Culture of Microorganisms (CNCM) in Paris under number 1-5893 in a culture medium suitable for the production of the bacterium excreting the exopolysaccharide in the culture medium; and - A second step of recovering the exopolysaccharide separated from the culture medium.

17. Method according to claim 16, characterized in that the exopolysaccharide present in the culture medium is separated from the solid biomass by solid-liquid phase separation.

18. Method according to claim 17, characterized in that the exopolysaccharide is separated from the culture medium by tangential filtration.

19. Method according to any one of claims 16 to 18, characterized by an additional step of cleaving the exopolysaccharide into at least one oligosaccharide according to any one of claims 11 to 15.

20. Method according to claim 19, characterized in that the cleavage is carried out by acid hydrolysis.

21. Mixture of oligosaccharides obtainable by the process according to claim 19 or 20 comprising mainly trisaccharides and / or heptasaccharides.

22. Mixture of oligosaccharides according to claim 21, characterized in that the trisaccharides have the following sequence: glucuronic acid, galacturonic acid and N-acetyl-quinovosamine (Glu-Gal-NAcQ).

23. Mixture of oligosaccharides according to claim 21, characterized in that the heptasaccharides have the following chain: glucuronic acid, galacturonic acid, N-acetyl-quinovosamine, rhamnose, glu-curonic acid, galacturonic acid and N-acetyl-quinovosamine (Glu-Gal-NAcQ-Rham-Glu-Gal-NAcQ).

24. Bacterial strain of Pseudoalteromonas sp. deposited in the National Collection of Culture of Microorganisms (CNCM) of Paris under number 1-5893.

25. Cosmetic or dermatological composition comprising an exopolysaccharide according to any one of claims 1 to 10 and a physiologically acceptable medium.

26. Cosmetic or dermatological composition comprising at least one oligosaccharide according to any one of claims 11 to 15 or a mixture of oligosaccharides according to any one of claims 21 to 23 and a physiologically acceptable medium.

27. ​​Use of the exopolysaccharide according to any one of claims 1 to 11 or of a composition containing it according to claim 26, for an application in the food, cosmetic, pharmaceutical, veterinary, agricultural, environmental or paint and coating fields.

28. Use of at least one oligosaccharide according to any one of claims 11 to 15 or a mixture of oligosaccharides according to any one of claims 21 to 23 or a composition containing them according to claim 26, for a non-therapeutic cosmetic treatment of the skin and its appendages.

29. Use according to claim 27, characterized in that the cosmetic treatment is topical.

30. Use according to any one of claims 27 or 29, characterized in that the cosmetic treatment is suitable for: - improving the radiance of the complexion; - improving the relief of the skin; - improving the firmness of the skin; and / or - improving the hydration of the skin.

31. Use according to any one of claims 27 to 29, characterized in that the cosmetic treatment is a treatment which protects the skin from aging and / or pollution, particularly atmospheric pollution.

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