FR3011248A1 - PROCESS FOR OBTAINING CLIOTED BIOACTIVE OLIGOSACCHARIDES, MICROORGANISM FOR ITS IMPLEMENTATION, OLIGOSACCHARIDES OBTAINED AND THEIR COSMETIC USE - Google Patents

Abstract

The method comprises providing a microorganism capable of producing at least one oside cleaving lyase enzyme, contacting polysaccharides of green algae, preferably of the genus Ulva, with said enzyme to cause said cleavage, and recovering said cleaved bioactive oligosaccharides. According to the invention, the microorganism is chosen from the group of bacteria of the Flavobacteriaceae family comprising the strain Jejuia pallidilutea, the strain Flavobacteriaceae bacterium, the strain Gaetbulibacter marinus and the strain referenced CNCM I-4797. The mixture of cleaved oligosaccharides obtained has a cosmetic activity on the cells of the skin, in particular anti-inflammatory, anti-radical, de-pigmenting effects, on molecules of the dermal extracellular matrix (such as collagen 1, collagen 4 and fibronectin), on molecules of the epidermal extracellular matrix (hyaluronic acid).

Description

DESCRIPTION The invention relates to a process for obtaining cleaved bioactive oligosaccharides obtained by enzymatic cleavage of polysaccharides derived from green algae, to a new microorganism for carrying out this process, to cleaved oligosaccharides obtained according to this method. process and their cosmetic use. The present invention thus relates in particular to the cosmetics industry which manufactures and / or uses products intended for the treatment of the skin, scalp, mucous membranes and integuments (such as hair, eyelashes, eyebrows, nails, hair) of mammals. animals or humans to improve their appearance and / or general condition.

The cosmetics industry is in growing demand for new products, in particular in demand for new active ingredients that are derived from plant material in order to be part of a sustainable development approach compared to a synthetic or chemical production route. Algae are thus an abundant source of raw material valued in many areas such as food, pharmacy and cosmetics.

In cosmetics, algae are already widely used but mainly for the gelling or thickening properties of their extracts which contain essentially polysaccharides. Polysaccharides also constitute a particularly interesting class of molecules, especially because they contain sequences comparable to certain bioactive oligomers. The Applicant has been particularly interested in green algae and more particularly in the genus ulva, and in the species ulva lactuca. It is known that polysaccharides derived from algae of the genus ulva contain, in particular, iduronic acid (IdopA) and rhamnose sulfate (Rhap3 sulfate) osidic units having an interesting potential for therapeutic or cosmetic applications (WO2009 / 016275). The Applicant is particularly interested in the two types of ulva polysaccharides that can be extracted from the walls of green algae of the genus ulva and containing such saccharide units: 1) the ulvan polysaccharide comprising two major osidic sequences, called "sequences of aldobiuronic acids ": a first (4) -13-D-GlcpA- (14) -aL-Rhap3 sulfate- (11) sequence and a second [4) -aL-IdopA- (14) - sequence. aL-Rhap3 sulfate- (11 ..

In these sequences, the "GlcpA" units correspond to glucuronic acid units. Inside this polysaccharide, one can also find sequences of the type "glucuronans", but in a minority. These sequences contain sequences of glucuronic acid units: [4] -13-D-GlcpA- (14) - (3-D-GlcpA- (11 ') 2) glucuronan polysaccharide which mainly comprises osidic sequences glucuronans containing chains of glucuronic acid units [4) - (3-D-GlcpA- (14) - (3-D-GlcpA- (11 ', as described above.

An enzyme of the class of lyases is able to cause the cleavage of the osidic bond linking the ose units according to a (3 elimination (or transelimination) reaction .An ulvan-lyase enzyme will be adapted to cleave more particularly the osidic bond between GlcpA and Rhap3 sulphate and between IdopA and Rhap3, while a glucuronan lyase enzyme will be adapted to cleave more particularly the glycosidic bond linking two GlcpA.The schematic mechanism of cleavage is as follows: This cleavage process reveals fragments of saccharide molecules oligosaccharides having a heterocycle having an ethylenic bond between carbon 4 and carbon 5 at the non-reducing terminal end thereof, this residue corresponding to a 4-deoxy- (hex-4-ene) pyranosyluronic acid. by 235 nm UV spectroscopy makes it possible to follow the appearance of this ethylene bond and the dosage of the products formed. cleavages consist of oligo-ulvans and oligo-glucuronans The proportion of each depends on the proportion of each of the glucuronan polysaccharides and ulvan starting contacted with the enzyme mixture.

Methods for cleaving ulva polysaccharides have been described comprising: providing a microorganism capable of producing an enzyme mixture of lyases; contacting polysaccharides extracted from algae of the genus ulva with said enzymatic mixture of lyases; to cause the cleavage of the saccharide bonds; and - recovering the cleaved oligosaccharides.

In the document WO2009 / 016275 mentioned above, a microorganism of the Ochrobactrum type trilled and deposited under the number I-3776 with the CNCM (National Collection of Microorganism Cultures, Institut Pasteur, Paris) is used to produce an enzymatic mixture having a double enzymatic activity, ulvan lyase and glucuronan lyase, and thus to obtain a mixture of oligo-glucuronans and oligo-ulvans having a degree of polymerization (DP) of less than 25 and in good yields for an industrial production. According to the present application, a DP corresponds to a diholoside unit (containing two monosaccharides). For example, a (4) - (3-D-GlcpA- (14) -αL-Rhap3 sulfate- (11n) sequence thus corresponds to a DP of 1, comprising a glucose / rhamnose diholoside unit The marine bacterium Nonlabens ulvanivorans of the family of flavobacteriaceae deposited under the number I-4324 at the CNCM has also been proposed to obtain such a saccharide cleavage The aim of the present invention is to propose an alternative method for obtaining mixtures of cleaved bioactive oligosaccharides cleaved enzymatic of polysaccharides derived from green algae H3C SO3Na O o H3C SO3Na O It thus proposes a process consisting in: - providing a microorganism capable of producing at least one oside cleavage lyase enzyme; - bringing said polysaccharides into contact with said enzyme to cause said cleavage; and - recovering said cleaved oligosaccharides, characterized in that said microorganism is selected from the group of bacteria of the family of Flavobacteriaceae comprising the strain Jejuia pallidilutea, the strain Flavobacteriaceae bacterium, the strain Gaetbulibacter marinus and the strain referenced CNCM I-4797. These different strains are related in terms of taxonomy. They make it possible to obtain cleaved oligosaccharides advantageously having particularly interesting cosmetic activities, as shown in particular by the results of in vitro tests given later as examples. The present invention more particularly covers the CNCM strain I-4797. It was filed in the name of the Applicant with the National Collection of Cultures of Microorganisms (CNCM) at the headquarters of the Pasteur Institute, 25 rue du Docteur Roux, F-75724 Paris cedex 15.

Preferably, the process according to the invention is carried out with this new strain CNCM I-4797. The strain Jejuia pallidilutea is in particular available from the DSMZ collection ("Deutsche Sammlung von Mikroorganismen und Zellkuturen GmbH", Inhoffen str 7B 38124 Braunschweig , Germany) under the reference DSMZ21165.

The Gaetbulibacter marinus strain is available in particular from the National Institute of Technology and Evaluation (NITE) collection, # 120, 2-5-8 Kazusakamatari, Kisarazu-shi, Chiba 292-0818, Japan), under the reference NBRC 102040 The microorganism according to the invention is capable of producing an ulvanase lyase cleavage enzyme and / or a glucuronan lyase enzyme, preferably both enzymes. Thus, the process according to the invention is preferably used to carry out the cleavage of sulphated polysaccharides derived from a green alga of the genus Ulva, in particular ulva lactuca, comprising sulphated polysaccharides of ulvans and / or glucuronans. high, such polysaccharides derived from green algae of the genus Ulva include: - aldobiuronic sequences [4) - (3-D-G1cpA- (14) -aL-Rhap3 sulfate- (11 and [4) -aL-IdopA - (14) - α-Rhap3 sulfate- (11) and - glucuronans [4) -13-D-G1cpA- (14) - (3-D-G1cpA- (11 'sequences, with in these sequences the GlcpA units corresponding to glucuronic acid units, Rhap3 sulfate rhamnose 3 sulfate units and IdopA units of iduronic acid The ulvan-lyase enzyme according to the invention is suitable for cleaving the saccharide bond between the GlcpA units and Rhap3 sulfate and between IdopA and Rhap3 sulfate, while the glucuronan lyase enzyme according to the invention is adapted to cleave the saccharide linkage. for two GlcpA units.

Preferably, according to the invention, the process is limited to cleaving ulvan polysaccharides previously cleared of glucuronan-type polysaccharides and thus mainly comprising aldobiuronic sequences, the resulting oligosaccharide mixture thus being enriched in oligo-ulvans.

The present invention also covers the cleaved bioactive oligosaccharides obtained according to the method of the invention, the present invention also covering non-therapeutic cosmetic use for the skin and its integuments of said cleaved oligosaccharides. The cosmetic treatment may be in particular a treatment chosen from a depigmenting treatment, anti-radical, hydrating (especially via the synthesis of hyaluronic acid of the epidermal matrix) or anti-aging via stimulation of the molecules of the extracellular dermal matrix (collagen 1, collagen 4 and fibronectin). According to the invention, it has been shown in particular that a mixture of cleaved oligosaccharides having a DP of between 2 and 10 makes it possible to obtain particularly interesting cosmetic activities as compared to other DPs. It goes without saying that other oligosaccharide mixture fractions can be obtained belonging to a different DP range depending on the operating conditions used for the process. In general, it will be sought to obtain oligosaccharides having a DP of less than 50, more preferably less than 20. According to the invention there is also provided a topical cosmetic composition comprising bioactive sulfated oligosaccharides according to the invention and a medium physiologically acceptable.

The degree of polymerization (DP) is measured by an ion chromatography method. The average weight of the fraction is 6 kD measured by CPC / DEDL (centrifugal partition chromatography) with a sulfation rate of 13 to 20%. According to other advantageous features, the oligosaccharides may be used in combination with one or more additional active ingredients, advantageously making it possible to offer a wider range of cosmetic properties. The additional active ingredients are used either as reinforcement of activity or in addition to activity. The additional active ingredients may for example be chosen from lightening active agents, anti-redness agents, stain removers, soothing agents, for the treatment of sensitive, reactive skin, UV sunscreens, moisturizing, moisturizing, exfoliating, smoothing and toning agents. firming, anti-aging, anti-wrinkles and fine lines, improving the mechanical and elastic properties, the radiance of the complexion, detoxifying active agents, anti-hair regrowth, acting on the cutaneous barrier, anti-acne, acting on the secretion of sebum , mattifying, unifying, anti-inflammatory, anti-oxidants, anti-free radicals, anti-glycation, eye contours (anti-dark circles and anti-pockets), promoting blood circulation, peptides, vitamins, etc. These active ingredients can be obtained from plant materials, such as plant extracts or vegetable or fermentation products. More specifically, the oligosaccharides of the present invention may advantageously be combined with oligo-glucuronans, especially the acetylated oligo-glucuronans described in the patent application WO2010067327 and sold in particular under the trade name SubliskinTM by Sederma. These are oligomeric compounds of D-glucuronic acid or D-glucuronate 13 (1-4) in the following formula (I): 0) O -0 0 OH

Preferably, each glucuronic acid cycle of formula (I) comprises a degree of acetylation of between 8.7 ± 0.5 and 9.2 ± 0, 5% by weight of O-CO-CH 3 groups relative to the weight of the glucuronic acid cycle and a degree of polymerization of between 18 and 19 ± 2. These oligomeric compounds act at the level of the elasticity of the dermis and the epidermis and increase the dermoepidermic cohesion in order to fight against the aging of the skin, the wrinkles, the wrinkles, the discontinuities of the visible and / or tactile skin, the loss of firmness, elasticity and tone, and to fight against the deformability of the cutaneous tissues. They also help moisturize the skin. Conventional actives such as vitamin B3 compounds, niacinamide or tocopherol, retinoid compounds such as retinol, hexamidine, α-lipoic acid, resveratrol or DHEA, peptides, especially N-acetyl- Tyr-Arg-O-hexadecyl, Pal-VGVAPG, Pal-KTTKS, Pal-GHK, Pal-KMO2K and Pal-GQPR can be combined with the oligosaccharides of the present invention. Other examples of additional ingredients are given later in the detailed description. The present invention also provides a method of topical treatment of the skin and its integuments, comprising the topical application of an effective amount of a composition according to the invention comprising oligosaccharides as defined above.

By "topical treatment" is meant an application that is intended to act at the place where it is applied: skin, mucosa, dander. Improvements in the appearance and the general condition of the skin of the mucous membranes and integuments can be obtained by topical application on a regular basis, for example daily.

The topical composition is preferably applied once daily for a period of at least one week, but may be applied for periods of 2, 4, 8 or 12 weeks. The topical composition is preferentially applied to the face and the neck, but may be applied to any part of skin requiring an aesthetic improvement, where the composition remains on the skin area to be treated, and preferably is not removed or rinsed from the skin. skin.

It should also be understood that, as used herein, the terms treat and treat include and include the reduction, amelioration, progression, alleviation, and / or elimination of dermatological effects, including aging. The compositions of the present invention as well as the methods are suitable for use in treating dermatological conditions of the skin in many areas of the skin, including but not limited to, the face, forehead, lips, neck, neck cleavage, arms, hands, body, legs, knees, feet, chest, back, buttocks, and others. One of the great advantages of the present invention resides in the possibility of being able to proceed, whenever necessary or desirable, to localized and selective "soft" treatments thanks to the mode of application by topical, non-invasive methods. An application can be made very localized using a syringe or a microcannula. The topical composition according to the invention is generally applied twice a day, morning and evening, but it can also be applied once a day, for a period of at least one week, and up to 12 weeks. By way of indication, for a cosmetic treatment of the figure, the European standard dosage in cream is 2.72 mg / cm 2 / day / person and for a cosmetic treatment of the body of 0.5 mg / cm 2 / day / person (according to data published by the European Scientific Committee for Consumer Safety (SCCS) "SCCS NOTES OF GUIDANCE FOR THE TESTING OF COSMETIC SUBSTANCES AND THEIR SAFETY EVALUATION 8TH REVISION", 11 December 2012)). According to other particularities, the cosmetic treatment method according to the invention may be associated with one or more other treatment methods aimed at the skin, such as, for example, treatments by light therapy, by heat or by aromatherapy. According to the invention, it is possible to propose devices with several compartments or kits intended for the implementation of the method described above, and which could include, by way of example, and without being limiting, in a first compartment containing a composition containing the oligosaccharides and in a second compartment a composition containing another active agent and / or excipient, the compositions contained in said first and second compartments being here considered as a combination composition for simultaneous, separate or spread over time especially in one of the treatments defined above. The present invention will be better understood in the light of the detailed description which follows. A. Preparation of oligosaccharides according to the invention Polysaccharide forming the starting substrate A mixture of polysaccharides contained in the walls of algae of the genus ulva lactuca is extracted while hot. After removal of the insolubles, the pH of the extract is lowered to 2 with dilute acid so that a precipitate including glucuronic acid residues is formed. The solution is then recovered, which is brought to a neutral pH close to 7, which is then purified, for example by ultrafiltration, and precipitated with alcohol. The glucuronan polysaccharide fraction is thus removed and an ulvan polysaccharide is obtained essentially including aldobiruonic sequences, the remaining glucuronane sequences being a minority and those integrated within the ulvan polysaccharide. The objective here is to subject only the ulvan polysaccharide to enzymatic cleavage in order essentially to recover oligo-ulvans (an enriched mixture of oligo-ulvans), but it goes without saying that the process can also alternatively be carried out on a mixture of ulvan and glucuronan polysaccharides The average molecular weight of the ulvan polysaccharide is determined by steric exclusion chromatography coupled to an in-line light scattering detector with a refractometer (SEC-MALLS Technology, for Size Exclusion). Chromatography Coupled to Laser-Light-Scattering Multi-angle, in English); this average mass is 6.105. Preparation of the bacterial strain From one of the bacterial strains according to the invention, here the strain referenced CNCM I-4797, an enzymatic substance having potentially a double enzymatic activity, ulvanolyase and glucuronan-lyase is prepared. The strain CNCM I-4797 according to the invention has the following characteristics: it is pigmented (orange / yellow color), gram negative and aerobic. The colonies on agar culture medium are creamy and opaque. In microscopy, medium-sized, non-motile bacilli are observed. The strain is cultured on a minimum mineral nutrient medium containing - NaCl: 22g; Na2SO4. 3.7 g KCl: 0.6 g; KBr: 0.1g; MgCl 2 · 6H 2 O: 10 g; CaCl2.2H2O: 2.94g; NaHCO3: 0.16 g; NaNO3: 25mg NaH3PO4,12H2O: 5mg and H20 QSP 1 liter; at a pH of 7.2. To this nutrient medium is added as substrate and carbon source ulvan polysaccharide prepared above (2g / L). The strain capable of degrading this ulvan polysaccharide is isolated on a petri dish containing the same nutritive medium supplemented with agar. After incubation, strain colonies which metabolize the substrate by degrading it in a mode of (3-elimination and which have been further developed are prepared.) Preparation of enzymatic activity The CNCM I-4797 strain is inoculated into a bioreactor 2 liter capacity, filled with 1.5 liters of mineral medium: NaCl: 22g, Na2SO4, 3.7g, KCl: 0.6g KBr: 0.1g, MgCl2.7H2O: 5mg NaNO3: 25mg, NaH3PO4.12H2O , H2O QSP 1 liter, at a pH of 7.2 and supplemented with yeast extract: 1 g / L, peptone: 5 g / L and ulvan polysaccharide (2 g / L) .The incubation is preferably carried out between 25 and 35 ° C. with stirring for an average duration of 48 hours During the incubation, the concentration of enzymatic mixture (ulvan-lyase and glucuronan-lyase) in the culture medium of the strain increases and consequently the enzymatic activity of the culture medium as well.The culture medium is then freed of bacteria after incubation or ammonium sulfate (60% w / v) is added to precipitate the proteins contained therein. The precipitated culture medium is then centrifuged and the protein pellet is recovered, which is then reintroduced into about 10 ml of a Tris / HC120 mM buffer solution at pH 7.5. Chromatography on anion exchange resin then makes it possible to recover exclusively the fraction corresponding to an enzymatic substance having predominantly ulvan lyase and minor glucuronan lyase activity. Enzymatic activity of 30 × 10 -3 μmol / ml / min is quantified by measuring the absorbance at 235 nm. Enzymatic cleavage The enzymatic substance obtained above is applied to a substrate solution containing 50 g of ulvan polysaccharide per 1 liter of enzyme solution. The degradation experiment is carried out at 20-30 ° C for an incubation period of 3 hours (theoretically ranging from 1 to 72 hours). The chromatographic profile according to SEC-MALLS technology described above makes it possible to follow the fraction of oligo-ulvans expected according to the invention (its average mass).

The analysis of the product resulting from the degradation by steric exclusion chromatography on Biogel P2 (detection by refractometry) shows a majority fraction analyzed by ESI-Q / TF type mass spectrometry (for electrospray-quadrupole-flight times). ). The oligosaccharides have a degree of polymerization DP of between 2 and 10, an average weight of 6 kD and a sulfation rate of 16.6%.

This process can be carried out on the other three bacterial strains according to the invention, namely Jejuia pallidilutea referenced DSMZ21165, the bacterial strain Flavobacteriaceae bacterium or the bacterial strain Gaetbulibacter marinus. B. In Vitro Tests The following various tests were carried out on the mixture of oligosaccharides obtained in point A above with the strain CNCM I-4797, to demonstrate the cosmetic activity. 1) Measurement of the anti-inflammatory effect on normal human fibroblasts (FHN): HHNs are cultivated until a confluent carpet is obtained. At this stage, they are put in contact with the products to be tested for 24 hours, then the carpets are irradiated (UVB 70 mJ / cm 2) and put back into contact with the products for 24 hours. The amounts of PGE2, IL-6 and IL-8 synthesized are measured in the culture supernatants by ELISA assay. 2) Measurement of the effect on the epidermal extracellular matrix (ECM): Human keratinocytes (HaCat) are cultured in plate for 24 hours. The cells are placed in contact or not with the test products for 3 days. The culture supernatants are removed and a determination of the amount of hyaluronic acid is carried out. Retinoic acid is used as a positive control. 3) Measurement of the anti-radical effect (intracellular ROS DCFH): FHNs are seeded in well plates. After 24 hours of attachment, the cells are brought into contact with the products for 24 hours. The products are eliminated and the carpets rinsed. The cells are loaded for 30 min with a probe "DCFH-DA" which becomes fluorescent in the presence of ROS 35 (oxygenated radical species). After rinsing, the cells are again brought into contact with the products and the stressor (H202 800um) for 2 hours. A fluorescence reading makes it possible to estimate the amount of ROS present in the cells. 4) Measurement of effect on dermal MEC (FHN): FHNs are cultured in plate for 24h. The cells are placed in contact or not with the test products for 3 days. The synthesis of collagen 1, collagen 4 and fibronectin is evaluated by the ELISA method. 5) Measurement of the depigmenting effect: Mouse melanomas (B16 cells) are seeded in plate in DMEMc medium ("DULBECCO'S Modified Eagle Medium") + 10 ° / 0SVF (fetal calf serum). After 24 hours of attachment, the cells are placed in contact with the products to be evaluated in the same medium for 48 hours. At the end of the contact, the culture media are eliminated and the total melanins are determined spectrophotometrically. The following tables 1 and 2 present the results of the various tests on the oligosaccharides according to the invention: Table 1: 1) Anti-inflammation 2) MEC 3) Anti-epidermal radical PGE2 IL-6 IL-8 Cleaved oligosaccharides according to the invention 0> 0> 0> 0 (hyaluronic acid)> 0 Table 2: 4) ECM dermal 5) Depigmentation Col 1 Fibronectin Col 4 Cleaved oligosaccharides according to the invention> 0> 0> 0 »0 These results show that the mixture of oligosaccharides according to the invention has a cosmetic activity. C. Preparation of a Composition According to the Invention By "physiologically acceptable medium" is meant according to the present invention, without being limiting, an aqueous or aqueous-alcoholic solution, a water-in-oil emulsion, an oil-in-oil emulsion. water, a microemulsion, an aqueous gel, an anhydrous gel, a serum, a dispersion of vesicles, a powder. "Physiologically acceptable" means for the present invention that the compositions are suitable for their topical use, in contact with the mucous membranes, superficial body growths (nails, hair, hair), scalp and mammalian skin and more particularly human, without risk of toxicity, incompatibility, instability, allergic response, and others. This "physiologically acceptable medium" forms what is conventionally called the excipient of the composition. The choice of the excipient of the composition is made according to the constraints related to the oligosaccharides (stability, solubilization, etc.) and, if appropriate, to the dosage form envisaged subsequently for the topical composition. The oligosaccharides may be incorporated into the composition by means of an aqueous solution, or may be solubilized by means of conventional physiologically acceptable solubilizers, for example and without limiting to this list: ethanol, propanol, isopropanol, propylene glycol, glycerin, butylene glycol, or polyethylene glycol or any combination thereof. It may also be advantageous to solubilize the oligosaccharides using emulsifiers. Preferably, according to the invention, an aqueous or hydro-alcoholic solution is used as the physiologically acceptable medium. According to the invention, the effective amount of oligosaccharides, i.e., their dosage, depends on various factors, such as the age, the condition of the patient's skin, etc. An effective amount means a non-toxic amount sufficient to achieve the desired effect that one skilled in the art is able to adjust. The topical compositions adapted to the present invention are generally prepared by usual methods well known to those skilled in the art. Such methods may involve mixing ingredients in one or more steps to achieve a uniform state, with or without heating, cooling, etc. The various galenic forms that may contain the oligosaccharides according to the invention are in any form namely creams, lotions, ointments milks or creams, gels, emulsions, dispersions, solutions, suspensions, cleaners, foundations, anhydrous preparations (sticks in particular lip balm, body and bath oils), shower and bath gels, shampoos and hair care lotions, milks or creams for the care of the skin or hair, lotions or make-up removers, lotions, milks or sunscreen creams, lotions, artificial tanning milks or creams, creams, mousses, gels or lotions of pre-shave, shaving, or aftershave, makeup, lipsticks, mascaras or nail polish, "essences" of skin, serums, adhesive or absorbent materials, patches, or powders, lotions, emollient milks or creams, sprays, body and bath oils, foundation bases, ointments, emulsions, colloids, compact suspensions or solids, pencils, spray formulations, blushes, reds, eyeliners, lipliners, lip glosses, face or body powders, styling foams or gels, nail conditioners, lip balms, skin conditioners, moisturizers , lacquers, soaps, exfoliants, astringents, depilatory permanent waving solutions, anti-dandruff formulations, antiperspirant or antiperspirant compositions, including sticks, "roll-on" deodorants, deodorants, sprays for nose and so on. These compositions may also be in the form of sticks for the lips intended either to color or avoid chapped skin, or eye makeup products or makeup and makeup for the face. The compositions according to the invention include beauty products, personal care products and pharmaceutical preparations. It is also possible to envisage a composition in the form of a foam or in the form of an aerosol composition also comprising a propellant under pressure. The oligosaccharides in the context of the present invention may be used in solution, dispersion, emulsion, paste or powder form, individually or in premix or be conveyed individually or in premix by vectors such as macrocapsules. , microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macroeponges, micro-sponges or nanoepongs, microemulsions or nanoemulsions, or adsorbed onto powdery organic polymers, talcs, bentonites, spores or exines and other inorganic or organic carriers. The oligosaccharides within the scope of the present invention may be used in any form, in a bound form, incorporated or adsorbed on macro-, micro-, and nanoparticles, or on macro-, micro- and nanocapsules, for the treatment textiles, natural or synthetic fibers, wool, and any material intended to come into contact with the skin and which may be used in clothing, night or daywear, handkerchiefs, or fabrics, in order to exert its cosmetic effect through this skin / textile contact and allow continuous topical delivery. Additional Ingredients CTFA ("International cosmetic ingredient dictionary & handbook" (13th Ed. 2010) published by "The Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, DC) describes a wide variety, without limitation, of cosmetic ingredients and pharmaceuticals customarily used in the skincare 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 oligosaccharides In addition, the nature of these additional ingredients must not unacceptably impair the benefits of the oligosaccharides of the invention.These additional ingredients may be synthetic or natural, such as plant extracts or from a process. Biofermentation Other skin care active ingredients pillar which are particularly useful in combination with the composition can be found in Sederma's commercial documentation and on the website www.sederma.fr. The following commercial active agents may also be mentioned as examples: betaine, glycerol, Actimoist Bio 2TM (Active Organics), AquaCacteenTM (Mibelle AG Cosmetics), AquaphylineTM (Silab), AquaregulKTM (Solabia), CarcilineTM (Greentech) ), CodiavelaneTM (Biotech Marine), DermafluxTM (Arch Chemicals, Inc.), Hydra'FlowTM (Sochibo), Hydromoist LTM (Symrise), RenovHyalTM (Soliance), SeamossTM (Biotech Marine), ArgirelineTM (trade name acetyl hexapeptide). 3 of Lipotec), spilanthol or an extract of Acmella oleracea known as Gatuline ExpressionTM, an extract of Boswellia serrata known as BoswellinTM, Deepaline PVBTM (Seppic), Syn-AKETM (Pentapharm), AmelioxTM, BioxiliftTM (Silab SubliskinTM (Sederma), VenuceaneTM (Sederma), Moist 24TM (Sederma), EssenskinTM (Sederma), JuvinityTM (Sederma), RevidratTM (Sederma), ResistemTM (Sederma), ChronodynTM (Sederma), KombuchkaTM (Sederma), ChromocareTM ( Sederma), Calmosensine ™ (Sederma), Glycokin factor STM (Sede) rma), Odawhite ™ (Sederma), Lumisphere ™ (Sederma) or mixtures thereof. Among the plant extracts that can be combined with the extract according to the invention, ivy extracts, for example climbing ivy (Hedera Helix), Bupleurum chinensis, Bupleurum Falcatum, arnica, may be mentioned in particular. (Arnica Montana L), rosemary (Rosmarinus officinalis / V), marigold (Calendula officinalis), sage (Salvia officinalis L), ginseng (Panax ginseng), Zingiber zerumbet sm., Ginko biloba, St. John's Wort ( Hyperycum Perforatum), European Butcher's Thistle (Ruscus aculeatus L), Ulmaria (Filipendula ulmaria L), Orthosiphon (Orthosiphon Stamincus Benth), Algae (Fucus Vesiculosus), Birch (Betula alba), Green Tea, cola nuts (Cola Nipida), horse chestnut, bamboo, Centella asiatica, heather, fucus, willow, piloselle, escin extracts, cangzhu extracts, chrysanthellum indicum chrysanthellum indicum extracts, plant extracts of the genus Armeniacea, Atractylodis Platicodon, Sinnomenum, Pharbitidis, Flemin gia, from Coleus as C. Forskohlii, C. blumei, C. esquirolii, C. scutellaroides, C. xanthantus and C. Barbatus, as an extract of raciness of Coleus barbatus, extracts of Ballote, Guioa, Davallia, Terminalia, Barringtonia , Trema, antirobia, cecropia, argania, dioscoreae such as Dioscorea opposita or Mexican, extracts of Ammi visnaga, Siegesbeckia, in particular Siegesbeckia orientalis, plant extracts of the Ericaceae family, in particular extracts of blueberries (Vaccinium angustifollium) from Arctostaphylos uva ursi, alpe vexa, plants containing sterols (in particular phytosterols), Manjistha (extract of plants of the genus Rubia, in particular Rubia Cordifolia), Guggal (extract of plants of the genus Commiphora, in particular Commiphora Mukul), cola extract, chamomile, purple clover, Piper methysticum (Kava Kava Sederma), Bacopa monieri (BacocalmineTM, Sederma) and sea whip, Glycyrrhiza glatira, mulberry, melaleuca (a tea), Larrea divaricata, Rabdosia rubescens, Euglena gracilis, Fibraurea recisa Hirudinea, Chaparral Sorghum, sunflower, Enantia chlorantha, Mitracarpe of the genus Spermacocea, Buchu barosma, Lawsonia inermis L ., Adiantium Capillus-Veneris L., Chelidonium malus, Luffa cylindrica, Japanese Mandarin (Citrus reticulata Blanco var. unshiu), Camelia sinensis, Imperata cylindrica, Glaucium Flavum, Cupressus Sempervirens, Polygonatum multiflorum, loveyly hemsleya, Sambucus Nigra, Phaseolus lunatus, Centaurium, Macrocystis Pyrifera, Turnera Diffusa, Anemarrhena asphodeloides, Portulaca pilosa, Humulus lupulus, Arabica coffee or Ilex Paraguariensis.

The compositions of the present invention may include peptides, including, but not limited to, di-, tri-, tetra-, penta- and hexapeptides and derivatives thereof. According to one particular embodiment, the concentration of the additional peptide in the composition varies between 1 × 10 -7% and 20%, preferably between 1 × 10 -6% and 10%, preferably between 1 × 10 -5% and 5%, by weight. . In the context of the present invention, the term "peptide" refers to peptides containing 10 amino acids or less, 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 synthetic peptides. It also refers to compositions which contain peptides and which are found in nature, and / or which are commercially available. Nonlimiting examples of dipeptides that may be used in the context of the present invention include Carnosine (beta-AH), YR, VW, NF, DF, KT, KC, CK, KP, KK or TT. Non-limiting examples of tripeptides include RKR, HGG, GHK, GKH, GGH, GHG, KFK, GKH, KPK, KMOK, KMO2K or KAvaK. Nonlimiting examples of tetrapeptides are RSRK, GQPR or KTFK. A non-limiting example of pentapeptide is KTTKS and hexapeptides GKTTKS and VGVAPG.

Other peptides that may be used in the context of the present invention may be chosen from, without this list being limitative: lipophilic derivatives of peptides, preferably palmitoyl derivatives, and complexes with the metal ions mentioned above (eg copper complex tripeptide HGG). Preferred dipeptides include, for example, N-Palmitoyl-beta-Ala-His, N-Acetyl-Tyr-Arghexadecylester (Calmosensine ™, Idealift ™, Sederma). Preferred tripeptides include Pal-GKH, Pal-GHK, the copper derivative of HGG (LaminTM, Sigma), lipospondin (N-Elaidoyl-KFK) and its conservative substitution analogues, N-Acetyl-RKR-NH2 ( Peptide CK +), NBiot-GHK (Sederma), Pal-KMO2K (Sederma Matrixyl Synthe6TM) and their derivatives. Tetrapeptide derivatives usable in the context of the present invention include, but are not limited to, NPal-GQPR (Sederma), useful pentapeptide derivatives are, but are not limited to, N-Pal-KTTKS (MATRIXYL ™, Sederma), N-Pal-Tyr-Gly-Gly-Phe-X with X being Met or Leu or their mixture. Useful hexapeptide derivatives include, but are not limited to: N-Pal-VGVAPG and derivatives. We can also mention the mixture Pal-GHK and Pal-GQPR (MatrixylTm 3000, Sederma). Preferred commercially available compositions containing a tripeptide or derivative include Biopeptide-CLTM, MaxilipTM or Sederma BiobustylTM. Preferred, commercially available sources of tetrapeptide sources include: RIGINTM, Eyeliss ™ Matrixyl ™ Reloaded and Matrixyl 3000 ™, which contain between 50 and 500 ppm of palmitoyl-GQPR and an excipient, provided by Sederma. The following commercial peptides may also be mentioned as additional active ingredients: VialoxTM, Syn-akeTM or Syn-CollTM (Pentapharm), Hydroxyprolisilane CNTM (Exsymol), Argireline TM, LeuphasylTM, AldenineTM, TrylgenTM, EyeserylTM, SerilesineTM or DecorinylTM (Lipotec) , CollaxylTM or QuintescineTM (Vincience), BONT-L-PeptideTM (Infinitec Activos), CytokinolTMLS (Serobiological Laboratories / Cognis), KollarenTM, IP2000TM or MelipreneTM (European Institute of Cell Biology), NeutrazenTM (Innovations), ECM-ProtectTm (Atrium Innovations) ), Timp-Peptide TM or ECM Modulin TM (Infinitec Activos).

Claims (14)

REVENDICATIONS1. A process for obtaining cleaved bioactive oligosaccharides obtained by enzymatic cleavage of polysaccharides derived from green algae, said process comprising: providing a microorganism capable of producing at least one oside-cleavage lyase enzyme; contacting said polysaccharides with said enzyme so as to cause said cleavage; and - recovering said cleaved oligosaccharides, characterized in that said microorganism is chosen from the group of bacteria of the family Flavobacteriaceae comprising the strain Jejuia pallidilutea, the strain Flavobacteriaceae bacterium, the strain Gaetbulibacter marinus and the strain referenced CNCM I- 4797. 2. Method according to claim 1, characterized in that said microorganism corresponds to the bacterial strain referenced CNCM I-4797. 3. Method according to claim 1 or 2, characterized in that said microorganism is capable of producing a cleavage enzyme ulvan lyase and / or a glucuronan lyase enzyme. 4. Method according to one of claims 1 to 3, characterized in that the polysaccharides are sulfated polysaccharides. 5. Process according to claim 4, characterized in that the sulphated polysaccharides are polysaccharides of ulvans and / or glucuronans 6. Method according to one of claims 1 to 5, characterized in that the green alga is of the genus Ulva. 7. Process according to claim 6, characterized in that the green alga is of the genus Ulva lactuca. 8. Micro-organism referenced CNCM I-4797 in particular for the implementation of the method according to one of claims 1 to 7. 9. Cleaved bioactive oligosaccharides obtained according to the process of any one of Claims 1 to 7. 10. Topical cosmetic composition comprising oligosaccharides according to claim 9 and a physiologically acceptable medium. 11. Non-therapeutic cosmetic use for the skin and its integuments of oligosaccharides according to claim 9 or a composition according to claim 10. 12. Non-therapeutic cosmetic use for the skin and its integuments of bioactive sulfated oligosaccharides obtained according to the method of claim 2 and 7, characterized in that the cleaved oligosaccharides have a degree of polymerization (DP) of between 2 and 10. 13. Use according to claim 11 or 12, characterized in that the cosmetic treatment is a treatment selected from a depigmenting, anti-radical, hydrating or anti-aging treatment via stimulation of the molecules of the dermal extracellular matrix. 14. Use according to one of claims 11 to 13, characterized in that the oligosaccharides are combined with a mixture of acetylated oligo-glucuronans oligomers Ducuronic acid or D-glucuronate 13 (1-4).