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Definitions

• This invention relates to an exopolysaccharide (EPS) excreted by bacterial strain CNCM 1-4150 of the Pseudoalteromonas sp species.
• This invention also relates to the use of this exopolysaccharide in cosmetic or dermopharmaceutical compositions for the treatment and/or care of the skin, mucous membranes, hair and/or nails.
• EPS exopolysaccharide
• the skin, mucous membranes, hair and/or nails constitute a physical barrier between the organism and its environment.
• the skin is composed of two tissues: the epidermis and the dermis.
• the epidermis is the outermost layer of the skin which is impermeable and therefore provides protection from external agents. It is a keratinized pluristratified epithelium which is continually renewing itself. Keratinocytes constitute the principal population of cells in the epidermis and are responsible for maintaining the epithelial structure due to their function as a barrier.
• the epidermis is composed of several cell layers; basal stratum which is the deepest layer connected to the dermis in the dermal-epidermal union and is composed of undifferentiated cells. Over time, these cells differentiate and migrate towards the surface of the epidermis, constituting the different layers.
• the uppermost layer formed is the stratum corneum which is composed by the corneocytes.
• the corneocytes are keratin, rich cells which are capable of retaining water, and are surrounded by a protein and a lipid shell. There are from 10 to 30 layers of stacked corneocytes, which are connected to each other by protein bridges called desmosomes.
• the resulting structure is a natural physical barrier of skin which retains water.
• Corneocytes are dead cells which are eliminated by desquamation, and which, in the absence of water, do not desquamate normally leading to a dense, dry and rough skin appearance.
• the loss of the superficial stratum caused by desquamation is compensated by the migration of cells from the basal stratum towards the surface of the epidermis. This is, therefore, a process of continual renewal of the skin which helps to keep it soft.
• the corneocytes contain a protein called filaggrin which binds to keratin proteins. Filaggrin is located in the outer part of the corneocytes, whilst keratin, which is capable of retaining water, remains in the inner part of the corneocytes.
• the dermis is the layer of skin located under the epidermis and firmly connected to it. It is an' elastic support tissue of mesodermal origin which is mainly constituted of fibroblasts and an extracellular matrix of fibrous proteins (collagen and elastin) and non-fibrous proteins (proteoglycans and glycoproteins).
• the dermis which is essentially rich in hyaluronic acid and polysaccharides, works as a reserve of water, retaining the water brought to it by the blood vessels. It stores water like a sponge and passes water to the epidermis when, is needed, together with other the nutritional substances the epidermis may also need. Therefore, the dermis plays a fundamental role in the development and differentiation of the epidermis..
• the fibroblasts and the extracellular matrix also influence on the mechanical properties of the skin, in particular, its elasticity, tone and firmness, as well as the skin's density.
• the skin can lose water in two ways: mainly through transpiration, which is an active phenomenon caused by the sweat glands to regulate the temperature of the skin, and also, although minimally, by passive evaporation of water through the epidermis. This passive evaporation or insensible water loss takes place with a kinetics that is the reflection of a balance between the water content of the epidermis and the relative humidity of the surroundings, and its measurement is the reflection of the integrity of the skin's barrier.
• insensible water loss is usually 5 g/m 2 /hour but in atopic children, and in areas of dry skin without eczema insensible water loss can reach 13 to 18 g/m 2 /hour.
• the integrity of the skin's barrier or the skin's barrier function also depends on the density of the corneum stratum.
• the corneum stratum has been compared to a brick wall in which the keratinocytes or corneocytes (bricks) are the essentially protein non- continuous portion, terminally differentiated, which are embedded in a continuous matrix of specialized lipids (mortar).
• the lipids provide the essential element of the barrier to water, and the corneocytes protect against continual abrasion by chemical or physical injuries.
• Hydration is an essential factor in the maintenance of the skin's youthfulness and vitality for any age group.
• the stratum corneum loses elasticity and experiments a sensation of tightness, a phenomenon which is usually referred to with the term "dry skin”.
• properly hydrated skin is soft, flexible and has a young, glowing look.
• Healthy skin is that which maintains ideal water concentration levels.
• the presence of water in the dermis and epidermis favors the group of regenerative mitotic reactions of the cutaneous cells, which contribute to the regeneration of our skin.
• An optimal water concentration is decisive for the flexibility of the skin and, as a consequence, for the prevention of the appearance of wrinkles caused by age and their treatment, and for the healing of small wounds.
• homeostasis of the skin can be affected by certain physiological factors (age, menopause, hormonal changes, lack of nourishment and lack of hydration, xerosis, etc.) or environmental factors (ultraviolet radiation, pollution, stress, hypoxia, infectious agents, dry weather conditions, irritants, etc.). These factors cause the decrease of an assimilation and fixation of water in the skin which quickly becomes obvious on the cutaneous surface through unmistakable signs such as dry skin or a tendency to irritation.
• hyaluronic acid an unsulfated glycosaminoglycan of the extracellular matrix formed by D-glucuronic acid and D-A/-acetylglucosamine.
• Hyaluronic acid is capable of retaining water in the skin, helping to maintain the skin more hydrated, elastic and with a more uniform cutaneous surface. The amount of hyaluronic acid which synthesizes the skin drastically reduces with age (Matuoka et al.
• Hyaluronic acid plays an important function in the prevention and decrease both wrinkles and expression lines; one of the more commonly employed strategies by the cosmetic and dermopharmaceutical industry for the treatment of wrinkles is the administration of hyaluronic acid both topically and subcutaneously due to its capacity of water absorption and therefore fill the wrinkle from inside the skin.
• Hyaluronic acid is found in the extracellular matrix of human and animal tissues, but it also exists in certain strains of bacteria such as those of the genus Streptococcus and Pasteurella, which produce it by emulating animal tissues as a way of protecting themselves against attack from the immune system of the animals they infect, as they are pathogenic microorganisms. Therefore, the production of hyaluronic acid is possible from the fermentation of bacteria which produce it naturally. In addition to this, it should be noted that its production is also possible through other genetically modified bacteria.
• exopolysaccharides described in the prior art which have been used for cosmetic and/or dermopharmaceutical purposes, such as the exopolysaccharide produced by a strain of bacteria of the genus Pseudomonas described in patent EP0534855 B1 which is used as a thickening, gelling and/or texturizing agent.
• patent application FR2871476A1 describes the GY785 strain of hydrothermal origin of the genus Alteromonas which produces an exopolysaccharide that can be used as a healing agent;
• patent EP0987010B1 describes an exopolysaccharide produced by a mesophilic bacterium of hydrothermal origin which improves the skin's defense system and
• patent application US2010/009931 describes the exopolysaccharide produced by a microalgae strain of the genus Porphyridium as a tensing agent, also improving the firmness, elasticity and tonicity of the skin.
• the American patent application US2009/069213A1 also describes the microalgae strain Porphyridium sp.
• Patent US6344346B1 also describes cosmetic compositions with hydrating properties caused by a polysaccharide of natural origin excreted by a bacterium of the genus Rhizobium.
• exopolysaccharide which has proven to have numerous advantageous properties for the skin is the exopolysaccharide described in application WO2009/127057, produced by strains of the bacterial species Staphylococcus epidermidis and Staphylococcus aureus. After applying a cosmetic composition of this exopolysaccharide the hydration and the morphology of the corneum extract improves, and the desquamation of the skin occurs.
• patent application JP2003-313131 should also be mentioned since it describes a polysaccharide sulfate produced by a strain of Alteromonas sp. SN-1009 (FERM BP- 5747) with anti-wrinkle properties.
• This invention relates to the cosmetic and/or dermopharmaceutical use of the exopolysaccharide excreted by the bacterial strain CNCM 1-4150 of the Pseudoalteromonas sp species.
• the inventors of this invention have found that the aforementioned exopolysaccharide is an alternative to hyaluronic acid which solves the problems caused by the lack of hydration of the skin, mucous membranes, hair and/or nails and evens out the skins surface.
• skin is understood to be the layers which comprise it, from the uppermost layer or stratum corneum to the lowermost layer or hypodermis, both inclusive. These layers are composed of different types of cells such as keratinocytes, fibroblasts, melanocytes and/or adipocytes among others.
• skin includes the scalp.
• care of the skin, mucous membranes, hair and/or nails comprises the prevention of disorders and/or diseases of the skin, mucous membranes, hair and/or nails.
• the term "aging” refers to the changes experienced by the skin with age (chronoaging) or through exposure to the sun (photoaging) or to environmental agents such as tobacco smoke, extreme climatic conditions of cold, heat, or wind, chemical contaminants or pollutants, and includes all the external visible and/or perceptible changes through touch, such as and not restricted to, the development of discontinuities on the skin such as wrinkles, fine lines, furrows, irregularities or roughness, increase in the size of pores, loss of elasticity, loss of firmness, loss of smoothness, loss of the capacity to recover from deformation, sagging of the skin such as sagging cheeks, the appearance of bags under the eyes or the appearance of a double chin, among others, changes to the color of the skin such as marks, reddening, bags under the eyes or the appearance of hyperpigmented areas such as age spots or freckles among others, anomalous differentiation, hyperkeratinization, elastosis, keratosis, hair loss, orange-peel skin, loss of
• photoaging groups together the set of processes due to the prolonged exposure of the skin to ultraviolet radiation which result in the premature aging of the skin, and present the same physical characteristics as aging, such as and not restricted to, flaccidity, sagging, changes to the color or irregularities in the pigmentation, abnormal and/or excessive keratinization.
• a first aspect of this invention relates to the exopolysaccharide of bacterial strain CNCM 1-4150 of Pseudoalteromonas sp. for the treatment and/or care of the skin, mucous membranes, hair and/or nails.
• the treatment and/or care of the skin, mucous membranes, hair and/or nails is a treatment and/or prevention of aging.
• the treatment and/or prevention of aging is a treatment and/or prevention of wrinkles on the skin and/or dryness of the skin.
• the treatment and/or care of the skin, mucous membranes, hair and/or nails is a treatment and/or care of conditions, disorders and/or diseases which are a result of the lack or decrease in hydration of the skin, mucous membranes, hair and/or nails.
• the conditions, disorders and/or diseases are selected from the group formed by dry skin, xerosis, hyperkeratosis, reactive hyperkeratosis, palmar and plantar hyperkeratosis, corns and calluses, actinic keratosis, non-actinic keratosis, atopic dermatitis, contact eczema, seborrheic dermatitis, dandruff, cradle cap on babies, acne, rosacea, nevus, ichthyosis, psoriasis, parakeratosis, pityriasis, lichen planus, palmoplantar keratoderma, chapped lips, vaginal dryness, ocular dryness, dry hair, brittle hair and nails.
• the treatment and/or care of the skin, mucous membranes, hair and/or nails is carried out by topical, transdermal, oral or parenteral application of the exopolysaccharide of the invention.
• parenteral includes nasal, auricular, ophthalmic, rectal, urethral, vaginal routes, subcutaneous, intradermal, intravascular injections, such as intravenous, intramuscular, intraocular, intravitreous, intracorneal, intraspinal, intramedullary, intracranial, intracervical, intracerebral, intrameningeal, intraarticular, intrahepatic, intrathoracic, intratracheal, intrathecal and intraperitoneal injections, as well as any another similar injection or infusion technique.
• the exopolysaccharide can be obtained through fermentation of the strain of Pseudoalteromonas sp. CNCM 1-4150 in a suitable culture medium, conventionally stirred and aired to synthesize and secrete the exopolysaccharide into the culture medium.
• Fermentation to produce the exopolysaccharide of this invention can be carried out in a medium stirred and aerated at a temperature between 20°C and 32°C, preferably at 29°C, the medium having a pH between 6.5 and 9, preferably around 7.5, adjusting it if necessary during fermentation.
• the duration of the fermentation is between 30 to 120 hours, preferably between 48 and 96 hours.
• exogenous sugars as a source of.carbon such as and not restricted to, galactose, glucose, mannose, amygdalin, cellobiose, maltose, starch, glycogen, lactose, mixtures thereof and/or extracts containing mixtures of these sugars.
• an exogenous supply of glucose of 2 to 40 g/L is provided, preferably from 15 to 25 g/L.
• Sugar incorporation methods to produce different polysaccharides are described in the prior art, for example and not restricted to, in documents: WO 98/38327, Raguenes et al. Int. J. Syst. Bad, 1997, 47:989-995 and Rougeaux er a/., Carboallos. Res., 1999, 322:40-45.
• mineral salts are provided for the fermentation culture of the bacterial strain CNCM 1-4150 of the species Pseudoalteromonas sp.
• they are selected from among salts which provide the ions Na + , K + , NH 4 + , Ca 2+ , Mg 2+ , P0 4 3" , S0 4 2" , CI " , C0 3 2” , or oligo elements such as Cu, Mn, Fe y Zn.
• the method of isolation and purification of the exopolysaccharide is carried out by the methods known by the person skilled in the art such as, centrifugation, filtration, ultrafiltration and dialysis.
• ultrafiltration and dialysis are carried out with a polyethersulfone membrane which retains molecules of a molecular weight greater than 100,000 Da.
• this invention relates to the native exopolysaccharide as well as to any chemical modification known by the person skilled in the art such sulfation as, methylation and/or acetylation, or the formation of exopolysaccharide- metal complexes.
• the molecular weight of the polysaccharide is modified by radical depolymerization resulting in a polymer with a molecular weight comprised between 100 and 800,000 Daltons, preferably a molecular weight of between 100 and 500,000 Daltons, and more preferably a molecular weight of between 100 and 100,000 Daltons.
• Depolymerization methods are known in the prior art, for example and not restricted to those described in Volpi et al. Anal. Biochem., 1992, 200:100-107.
• the exopolysaccharide excreted by the bacterial strain of the species of Pseudoalteromonas sp. CNCM 1-4150 is characterized by producing at least four different neutral monosaccharides and two acid monosaccharides.
• the neutral monosaccharides are preferably mannose, glucose, galactose and N- acetylglucosamine.
• the acid monosaccharides are preferably glucuronic acid and galacturonic acid.
• the exopolysaccharide of this invention produces a composition in weight of 3% to 12% of mannose, 12% to 34% of glucose, 12% to 34% of glucuronic acid, 2% to 20% of galacturonic acid, 12% to 34% of galactose and 2% to 18% of W-acetylglucosamine, with the condition that the sum of the percentages does not exceed 100%.
• the exopolysaccharide produces a composition in weight of 4% to 10% of mannose, 17% to 29% of glucose, 17% to 29% of glucuronic acid, 4% to 18% of galacturonic acid, 17% to 29% of galactose and 4% to 14% of W-acetylglucosamine. Even more preferably, the exopolysaccharide produces a composition in weight of 5% to 9% of mannose, 20% to 26% of glucose, 20% to 26% of glucuronic acid, 9% to 15% of galacturonic acid, 20% to 26% of galactose and 7% to 12% of /V-acetylglucosamine.
• the exopolysaccharide in addition contains rhamnose.
• a second aspect of this invention relates to a cosmetic or dermopharmaceutical composition characterized in that it comprises a cosmetically or dermopharmaceutically effective quantity of the exopolysaccharide of this invention and at least one excipient, adjuvant and/or cosmetically and/or dermopharmaceutically acceptable ingredient.
• the cosmetically or dermopharmaceutically effective quantity of the exopolysaccharide in the composition of the invention to be administered, as well as its dosage will depend on numerous factors, including age, condition of the patient, the nature or severity of the condition, disorder or disease to be treated and/or cared for, the route and frequency of administration and the nature, in particular, of the exopolysaccharides to be used.
• Cosmetically or dermopharmaceutically effective is understood to be a non-toxic but sufficient quantity of the exopolysaccharide to provide the desired effect.
• the exopolysaccharide of the invention is used in the cosmetic or dermopharmaceutical composition of this invention at cosmetically or dermopharmaceutically effective concentrations to achieve the desired effect; preferably, with regards to the total weight of the composition, between 0.00000001 % (in weight) and 20% (in weight); preferably between 0.000001% (in weight) and 20% (in weight), more preferably between 0.0001% (in weight) and 10% (in weight) and even more preferably between 0.0001% (in weight) and 5% (in weight).
• the exopolysaccharide of the invention can also be incorporated into cosmetic and/or dermopharmaceutical delivery systems and/or sustained release systems.
• delivery systems relates to a cosmetically and/or dermopharmaceutically acceptable carrier such as a diluent, adjuvant, excipient, vehicle or additives with which the exopolysaccharide of the invention is administered.
• a cosmetically and/or dermopharmaceutically acceptable carrier such as a diluent, adjuvant, excipient, vehicle or additives with which the exopolysaccharide of the invention is administered.
• These delivery systems are well known in the prior art and can be used for, example, to improve the definitive formulation with regards to organoleptic properties, penetration of the skin and the bioavailability of the active ingredient.
• These cosmetic and/or dermopharmaceutical vehicles can be liquids, such as water, oils or surfactants, including those of petroleum, animal, plant or synthetic origin, such as and not restricted to, peanut oil, soybean oil, mineral oil, sesame oil, castor oil, polysorbates, sorbitan esters, ether sulfates, sulfates, betaines, glycosides, maltosides, fatty alcohols, nonoxynols, poloxamers, polyoxyethylenes, polyethylene glycols, dextrose, glycerol, digitonin and similar.
• sustained release is used in a conventional sense relating to a delivery system of a compound which provides the gradual release of this compound during a period of time and preferably, although not necessarily, with relatively constant compound release levels over a period of time.
• Examples of delivery or sustained release systems are liposomes, mixed liposomes, oleosomes, niosomes, ethosomes, milliparticles, microparticles, nanoparticles and solid lipid nanoparticles, nanostructured lipid supports, sponges, cyclodextrins, vesicles, micelles, mixed micelles of surfactants, surfactant-phospholipid mixed micelles, millispheres, microspheres and nanospheres, lipospheres, millicapsules, microcapsules and nanocapsules, as well as microemulsions and nanoemulsions, which can be added to achieve a greater penetration of the active principle and/or improve its pharmacokinetic and pharmacodynamic properties.
• Preferred delivery or sustained release systems are liposomes, surfactant-phospholipid mixed micelles and microemulsions, more preferably water-in-oil microemulsions with an internal reverse micelle structure.
• the sustained release systems can be prepared by methods known in the prior art, and the compositions which contain them can be administered, for example, by topical or transdermal administration, including adhesive patches, non-adhesive patches, occlusive patches and microelectric patches, or by systemic administration, such as and not restricted to, orally or parenterally, including nasal, rectal or subcutaneous implantation or injection, or direct implantation or injection into a specific body part, and preferably should release a relatively constant quantity of the exopolysaccharide of the invention.
• topical or transdermal administration including adhesive patches, non-adhesive patches, occlusive patches and microelectric patches
• systemic administration such as and not restricted to, orally or parenterally, including nasal, rectal or subcutaneous implantation or injection, or direct implantation or injection into a specific body part, and preferably should release a relatively constant quantity of the exopolysaccharide of the invention.
• the amount of exopolysaccharide contained in the sustained release system will depend, for example, on where the composition is to be administered, the kinetics and duration of the release of the exopolysaccharide of the invention, as well as the nature of the condition, disorder and/or disease to be treated and/or cared for.
• composition containing the exopolysaccharide of this invention can also be adsorbed on solid organic polymers or solid mineral supports, such as and not restricted to, talc, bentonite, silica, starch or maltodextrin among others.
• compositions containing the exopolysaccharide of the invention can also be incorporated into fabrics, non-woven fabrics or medical devices which are in direct contact with the skin, thus freeing the exopolysaccharide of the invention whether by biodegradation of the binding system to the fabric, non-woven fabric or medical device, or due to the friction between them and the body, due to body moisture, the skin's pH or body temperature.
• the fabrics and non-woven fabrics can be used for making garments that are in direct contact with the body.
• the preferred fabrics, non-woven fabrics, garments and medical devices are bandages, gauzes, t-shirts, socks, tights, underwear, girdles, gloves, diapers, sanitary napkins, dressings, bedspreads, flannels, adhesive patches, non-adhesive patches, occlusive patches, microelectric patches and/or face masks.
• compositions containing the exopolysaccharide of this invention can be used in different types of compositions of topical or transdermal application, optionally including cosmetically and/or dermopharmaceutically acceptable excipients necessary for formulating the desired administration form.
• compositions of topical or transdermal application can be produced in any solid, liquid or semisolid formulation.
• these compositions of topical or transdermal application are, for example and not restricted to, creams, multiple emulsions, such as and not restricted to, oil and/or silicone in water emulsions, water-in-oil and/or silicone emulsions, water/oil/water or water/silicone/water type emulsions, and oil/water/oil or silicone/water/silicone type emulsions, microemulsions, emulsions and/or solutions, liquid crystals, anhydrous compositions, aqueous dispersions, oils, milks, balsams, foams, aqueous or oily lotions, aqueous or oily gels, cream, hydroalcoholic solutions, hydroglycolic solutions, hydrogels, liniments, sera, soaps, shampoos, conditioners, face masks, hairsprays, serums, polysaccharide films,
• formulations are topically or transdermal ⁇ applied on local areas of the skin, mucous membranes, hair and/or nails and can be incorporated using techniques known by the person skilled in the art into different types of solid accessories, such as and not restricted to, bandages, gauzes, t-shirts, socks, tights, underwear, girdles, gloves, diapers, sanitary napkins, dressings, bedspreads, flannels, adhesive patches, non-adhesive patches, occlusive patches, microelectric patches and/or face masks, or they can be incorporated into different make-up products such as make-up foundation, such as fluid foundations and compact foundations, lotions or make-up removers, make-up removal milks, under-eye concealers, eye shadows, lipsticks, lip protectors, lip gloss and powders among others.
• make-up foundation such as fluid foundations and compact foundations, lotions or make-up removers, make-up removal milks, under-eye concealers, eye shadows, lipsticks, lip protectors
• compositions of the invention may include agents which increase the percutaneous absorption of the exopolysaccharide of this invention, for example and not restricted to, dimethyl sulfoxide, dimethylacetamide, dimethylformamide, surfactants, azone (1-dodecylazacycloheptane-2-one), alcohol, urea, ethoxydiglycol, acetone, propylene glycol or polyethylene glycol, among others.
• agents which increase the percutaneous absorption of the exopolysaccharide of this invention for example and not restricted to, dimethyl sulfoxide, dimethylacetamide, dimethylformamide, surfactants, azone (1-dodecylazacycloheptane-2-one), alcohol, urea, ethoxydiglycol, acetone, propylene glycol or polyethylene glycol, among others.
• compositions of this invention can be applied to local areas to be treated by means of iontophoresis, sonophoresis, electroporation, microelectric patches, mechanical pressure, osmotic pressure gradient, occlusive cure, microinjections or needle-free injections by means of pressure, such as injections by oxygen pressure, or any combination thereof, to achieve a greater penetration of the exopolysaccharide of the invention.
• the application area will be determined by the nature of the condition, disorder and/or disease to be treated and/or cared for.
• the cosmetic or dermopharmaceutical compositions containing the exopolysaccharide of this invention can be used in different types of formulations for oral administration, preferably in the form of oral cosmetics or medication, such as and not restricted to, capsules, including gelatin capsules, soft capsules, hard capsules, tablets, including sugar coated tablets, powders, granules, chewing gum, solutions, suspensions, emulsions, syrups, polysaccharide films, jellies or gelatins, and any other form known by the person skilled in the art.
• the exopolysaccharide of the invention can be incorporated into any form of functional food or fortified food, such as and not restricted to, dietary bars or compact or non-compact powders.
• exopolysaccharide of this invention can be formulated with common excipients and adjuvants for oral compositions or food supplements, such as and not restricted to, fat components, aqueous components, humectants, preservatives, texturizing agents, flavors, aromas, antioxidants and dyes common in the food industry.
• compositions containing the exopolysaccharide of the invention can also be administered by topical or transdermal route, as well as by any other appropriate route, for example oral or parenteral route, for which they will include the pharmaceutically acceptable excipients necessary for the formulation of the desired administration of the exopolysaccharide.
• adjuvants and/or ingredients contained in the cosmetically or dermopharmaceutically acceptable compositions described in this invention are additional ingredients commonly used in compositions for the treatment and/or care of the skin, mucous membranes, hair and/or nails such as and not restricted to, agents inhibiting acetylcholine receptor clustering, muscle contraction inhibiting agents, anticholinergic agents, elastase inhibiting agents, matrix metalloprotease inhibiting agents, melanin synthesis stimulating or inhibiting agents, whitening or depigmenting agents, propigmenting agents, self-tanning agents, antiaging agents, NO-synthase inhibiting agents, 5a-reductase inhibiting agents, lysyl- and/or prolyl hydroxylase inhibiting agents, antioxidants, free radical scavengers and/or agents against atmospheric pollution, reactive carbonyl species scavengers, anti- glycation agents, antihistamine agents
• these additional ingredients should not unacceptably alter the benefits of the exopolysaccharide of this invention.
• the nature of these additional ingredients can be synthetic or natural, such as vegetable extracts, or obtained by a biofermentation process or from a combination of a synthetic process and a biotechnological process. Additional examples can be found in the CTFA International Cosmetic Ingredient Dictionary & Handbook, 12th Edition (2008).
• biotechnological process is understood to be any process which produces the active ingredient, or part of it, in an organism, or in one part of it.
• the anti-wrinkle and/or antiaging agent is selected, for example and not restricted to, from the group formed by extracts of Vitis vinifera, Rosa canina, Curcuma longa, Iris pallida, Theobroma cacao, Ginkgo biloba, Leontopodium Alpinum or Dunaliella salina, Matrixyl ® [INCI: Palmitoyl Pentapeptide-4], Matrixyl 3000 ® [INCI: PalmitoyI Tetrapeptide-7, PalmitoyI Oligopeptide], Essenskin [INCI: Calcium Hydroxymethionine], Renovage [INCI: Teprenone] or Dermaxyl ® [INCI: PalmitoyI Oligopeptide] marketed by Sederma/Croda, Vialox ® [INCI: Pentapeptide-3], Syn ® -Ake ® [INCI: Dipeptide Diaminobutyroyl Benzylamide Diacetate], Syn ®
• the humectant or moisture retaining substance, moisturizer or emollient is selected, for example and not restricted to, from the group formed by polyols and polyethers such as glycerin, ethylhexylglycerin, caprylyl glycol, pentylene glycol, butylene glycol, propylene glycol and its derivatives, triethylene glycol, polyethylene glycol, Glycereth-26, Sorbeth-30; panthenol; pyroglutamic acid and its salts and derivatives; amino acids, such as serine, proline, alanine, glutamate or arginine; ectoin and its derivatives; A/-(2-hydroxyethyl)acetamide; /V-lauryl-pyrrolidone carboxylic acid; A/-lauryl-L-lysine; /V-alpha-benzoyl-L-arginine; urea; creatine; a-
• the agent stimulating healing, coadjuvant healing agent, agent stimulating reepithelialization and/or coadjuvant reepithelialization agent is selected, for example and not restricted to, the group formed by extracts of Aristoloquia clematis, Centella asiatica, Rosa moschata, Echinacea angustifolia, Symphytum officinale, Equisetum arvense, Hypericum perforatum, Mimosa tenuiflora, Persea gratisima, Prunus africanum, Tormentilla erectea, Aloe vera, Polyplant ® Epithelizing [INCI: Calendula Officinalis, Hypericum Perforatum, Chamomilla Recutita, Rosmarinus Officinalis] marketed by Provital, Cytokinol ® LS 9028 [INCI: Hydrolyzed Casein, Hydrolyzed Yeast Protein, Lysine HCI] marketed by Laboratories
• the agent stimulating the synthesis of dermal or epidermal macromolecules is selected, for example and not restricted to, from the group formed by collagen synthesis-stimulating agent, elastin synthesis-stimulation agent, decorin synthesis-stimulation agent, laminin synthesis-stimulation agent, chaperone synthesis- stimulating agent, sirtuin synthesis-stimulating agent, hyaluronic acid synthesis- stimulating agent, aquaporin synthesis-stimulating agent, fibronectin synthesis- stimulating agent, agents that inhibit collagen degradation, agents that inhibit serine proteases such as leukocyte elastase or cathepsin G, agents stimulating fibroblast proliferation, agents stimulating adipocyte proliferation, agents stimulating adipocyte differentiation, glycosaminoglycan synthesis-stimulating agents, and DNA repairing agents and/or DNA protecting agents, such as and not restricted to extracts of Centella asiatica, Saccharomyces cerevisiae, Solanum tuberosum, Rosmarin
• the agent inhibiting elastin degradation is selected, for example and not restricted to, from the group formed by Elhibin ® [INCI: Glycine Soja (Soybean) Protein], Preregen ® [INCI: Glycine Soja (soybean) Protein, Oxido Reductases] or Regu ® -Age [INCLHydrolyzed Rice Bran Protein, Glycine Soja (Soybean) Protein, Oxido Reductases] marketed by Pentapharm/DSM, Juvenesce [INCI: Ethoxydiglicol and Caprylic Triglycerid, Retinol, Ursolic Acid, Phytonadione, llomastat], MicromerolTM [INCI: Pyrus Malus Extract], Heather Extract [INCI: Calluna Vulgaris Extract], Extracellium ® [INCI: Hydrolyzed Potato Protein] or FlavagrumTM PEG [INCI: PEG-6 Isostearate,
• the matrix metalloproteinase inhibitory agent is selected, for example and not restricted to, from the group formed by ursolic acid, isoflavones such as genistein, quercetin, carotenoids, lycopene, soya extract, cranberry extract, rosemary extract, extract of Trifolium pratense (red clover), extract of Phormium tenax (New Zealand flax), kakkon-to extract, sage extract, retinoi and its derivatives, retinoic acid and its derivatives, sapogenins such as diosgenin, hecogenin, smilagenin, sarsapogenin, tigogenin, yamogenin and yucagenin, among others, Collalift ® [INCI: Hydrolyzed Malt Extract ], Juvenesce [INCI: Ethoxydiglicol and Caprylic Triglyceride, Retinoi, Ursolic Acid, Phytonadione,
• the firming and/or redensifying agent is selected, for example and not restricted to, from the group formed by extracts of Malpighia punicitolia, Cynara scolymus, Gossypium herbaceum.
• the desquamating agent and/or keratolytic agent and/or exfoliating agent is selected, for example and not restricted to, from the group formed by hydroxyacids and their derivatives, ?-hydroxyacids, in particular salicylic acid and its derivatives, or gentisic acid; ⁇ -hydroxyacids and its salts, such as glycolic acid, ammonium glycolate, lactic acid, 2-hydroxyoctanoic acid, ⁇ -hydroxycaprylic acid, mandelic acid, citric acid, malic acid or tartaric acid; a- and ?-hydroxybutyric acids; polyhydroxy acids such as gluconic acid, glucuronic acid or saccharic acid; keto acids such as pyruvic acid, glyoxylic acid; carboxylic pyrrolidine acid; cysteic acid and derivatives; aldobionic acids; azelaic acid and its derivatives such as azeloyl diglycinate; ascorbic acid and its derivatives such as 6-O-
• a third aspect of this invention refers to the use of the exopolysaccharide of the invention in the preparation of a cosmetic or dermopharmaceutical composition for the treatment and/or care of the skin, mucous membranes, hair and/or nails.
• this invention refers to the use of the exopolysaccharide in the preparation of a cosmetic or dermopharmaceutical composition for the treatment and/or prevention of aging.
• the treatment and/or prevention of aging is a treatment and/or prevention of wrinkles on the skin and/or dry skin.
• this invention refers to the use of the exopolysaccharide in the preparation of a cosmetic .
• the conditions, disorders and/or diseases are selected from the group formed by dry skin, xerosis, hyperkeratosis, hyperkeratosis response, palmar and plantar hyperkeratosis, corns and calluses, actinic keratosis, non-actinic keratosis, atopic dermatitis, contact eczema, seborrheic dermatitis, dandruff, cradle cap on babies, acne, rosacea, nevus, ichthyosis, psoriasis, parakeratosis, pityriasis, lichen planus, palmoplantar keratoderma, chapped lips, vaginal dryness, ocular dryness, dry
• cosmetic or dermopharmaceutical compositions for the treatment and/or care of the skin, mucous membranes, hair and/or nails include creams, multiple emulsions, such as and not restricted to, oil and/or silicone in water emulsions, water- in-oil and/or silicone emulsions, water/oil/water or water/silicone/water type emulsions, and oil/water/oil or silicone/water/silicone type emulsions, anhydrous compositions, aqueous dispersions, oils, milks, balsams, foams, lotions, gels, cream gels, hydroalcoholic solutions, hydroglycolic solutions, hydrogels, liniments, sera, soaps, shampoos, conditioners, serums, polysaccharide films, ointments, mousses, pomades, powders, bars, pencils and sprays or aerosols (sprays), including leave-on and rinse- off formulations, bandages, gauzes, t
• compositions containing the exopolysaccharide of this invention can also be incorporated into products for the treatment, care and/or cleaning of nails and cuticles such as nail varnishes, nail varnish remover and cuticle remover lotions, among others.
• the compositions containing the exopolysaccharide of this invention can be applied to the skin, mucous membranes, hair and/or nails or can be administered orally or parenterally depending on the requirements to treat and/or care for a condition, disorder and/or disease.
• compositions of this invention can be applied to the skin by means of iontophoresis, sonophoresis, electroporation, microelectric patches, mechanical pressure, osmotic pressure gradient, occlusive cure, microinjections or needle-free injections by means of pressure, such as injections by oxygen pressure, or any combination thereof, to achieve a greater penetration of the exopolysaccharide of the invention.
• An additional aspect of this invention refers to a method of treatment and/or care of the skin, mucous membranes, hair and/or nails which comprises the administration of a cosmetically and/or dermopharmaceutically effective quantity of the exopolysaccharide, preferably in the form of a cosmetic or dermopharmaceutical composition containing it.
• Another additional aspect of this invention refers to a method for the treatment and/or care of any conditions, disorders and/or diseases of mammals, preferably of humans, which are a consequence of a lack or decrease in the hydration of the skin, mucous membranes, hair and/or nails, which comprises the administration of an effective quantity of the exopolysaccharide, preferably in the form of a cosmetic or dermopharmaceutical composition containing them.
• the conditions, disorders and/or diseases which are a consequence of a lack or decrease in the hydration of the skin, mucous membranes, hair and/or nails are selected from the group formed by dry skin, xerosis, hyperkeratosis, hyperkeratosis response, palmar and plantar hyperkeratosis, corns and calluses, actinic keratosis, non-actinic keratosis, atopic dermatitis, contact eczema, seborrheic dermatitis, dandruff, cradle cap on babies, acne, rosacea, nevus, ichthyosis, psoriasis, parakeratosis, pityriasis, lichen planus, palmoplantar keratoderma, chapped lips, vaginal dryness, ocular dryness, dry hair, brittle hair and nails.
• this invention refers to the treatment and/or care which reduces, delays and/or prevents the signs of aging and which comprises the administration of an effective quantity of the exopolysaccharide, preferably in the form of a cosmetic or dermopharmaceutical composition containing it.
• the treatment and/or care which reduces, delays and/or prevents the signs of aging is a treatment and/or prevention of wrinkles on the skin and/or dryness of the skin.
• the treatment and/or care of this invention is carried out by topicai or transdermal application, preferably, the topicai or transdermal application is carried out by means of iontophoresis, sonophoresis, electroporation, mechanical pressure, osmotic pressure gradient, occlusive cure, microinjections, with needle-free injections by means of pressure, microelectric patches or any combination thereof.
• the treatment and/or care is carried out by oral administration.
• the treatment and/or care is carried out by parenteral application.
• the frequency of the application or administration can vary widely, depending on the needs of each subject and severity of the condition, disorder or disease to be treated or cared for, suggesting a range of application or administration from once per month to tenth times per day, preferably from once per week to four times per day, more preferably from three times per week to three times per day, even more preferably once or twice per day.
• Figure 1 shows a comparative study of water retention between the exopolysaccharide of this invention and the hyaluronic acid using the dynamic vapor sorption technique.
• Example 1 Preparation and isolation of the exopolysaccharide secreted by strain CNCM 1-4150 corresponding to the species Pseudoalteromonas sp. a) Method of cultivation of strain CNCM 1-4150 corresponding to the species Pseudoalteromonas.
• Strain CNCM 1-4150 was cultivated in a fermenter, at 29°C and at a pH of 7.5, whose broth contained 2216E medium (ZoBell C.E. J. Mar. Res., 1941 , 4:42.) enriched with glucose (20 g/l).
• An inoculum was prepared with 10% (v/v) of a previous crop and the duration of the fermentation was extended to 72 hours. The speed of aeration and stirring was 2 wm and 250 rpm, respectively.
• the bacteria were separated from the broth by centrifugation at 12,000 g for 45 mins.
• the polysaccharide was purified with distilled water by ultrafiltration with a polyethersulfone membrane for polysaccharides of over 100 KDa in molecular weight. Once purified, the polysaccharide was depolymerized by radical depolymerization (Volpi N. et al. Anal. Biochem., 1992, 200:100-107) resulting in a polymer with a molecular weight between 3,000 and 40,000 Da.
• Example 2 Physical-chemical characterization of the exopolysaccharide produced by bacterial strain CNCMI-4150 corresponding to the species Pseudoalteromonas sp. a) Chemical analysis
• the content of neutral and acid monosaccharides of the exopolysaccharide obtained was determined according to that described in example 1 by hydrolysis and chromatography of gases according to the method described by Kamerling et al. Biochem. J, 1975 151 :491-495, and modified by Montreuil et al. in 1986, Glycoproteins. In Carbohydrate analysis: a practical approach. Eds Chaplin et Kennedy, I.R.L Press, Oxford, Washington D.C., pp143-204.
• Example 3 Preparation of a cosmetic composition of the exopolysaccharide excreted by bacterial strain CNCM 1-4150.
• phase A ingredients water [INCI: Water (Aqua)], Phenonip [INCI: Phenoxyethanol, Methylparaben, Ethylparaben, Butylparaben, Propylparaben, Isobutylparaben], Abiol [INCI: Imidazolidinyl Urea] and Propylene Glycol USP/EP [INCI: Propylene Glycol] (phase A ingredients).
• the mixture of ingredients in phase A was subjected to constant stirring and subsequently Carbopol ETD 2020 [INCI: Acrylates/C10-30 Alkyl Acrylate Crosspolymer] was added (phase B). The resulting mixture was heated in a microwave to 65°C.
• Lipomulse 165 Glyceryl Stearate, PEG-100 Stearate
• Alcohol CO-1695 INCI: Cetyl Alcohol
• Edenor L2SM INCI: Stearic Acid, Palmitic Acid
• Caprylic Capric Triglycerides INCI: Caprylic/Capric Triglyceride
• Massocare HD Isohexadecane
• Example 4 Preparation of a cosmetic composition of the exopolysaccharide excreted by bacterial strain CNCM 1-4150.
• the cosmetic composition of this example was prepared following the instructions for the preparation of the composition of example 3 with the same ingredients, but using the quantities in table 2.
• Example 5 Comparative study of water retention between the hyaluronic acid and the exopolysaccharide of strain CNCM 1-4150.
• the experiments were carried out with the dynamic vapor sorption technique (DVS), with a TA Instruments Q5000 SA thermogravimetric analyzer (TGA), treating the values obtained with the Universal Analysis 2000 version 4.5A (TA Instruments).
• the protocol used considers an initial equilibrium step at 60°C, establishing humidity at 0.0%, and a subsequent equilibrium at 33°C, from which the relative humidity is begun to be raised in 10% stages. Once 95% has been reached successive stages of lowering the relative humidity are carried out. Throughout the whole period of variation of the relative humidity the weight of the exopolysaccharide is recorded. Afterwards the same experiment was carried out under the same conditions with hyaluronic acid.
• Example 6 In vivo study of skin hydration.
• the measurements of this study were carried out in a bioclimatic room (24 ⁇ 2 °C; 50 ⁇ 10% relative humidity) with the purpose of maintaining the temperature and humidity constant during the measuring.
• the measurements of skin hydration were carried out on the cheeks using a Corneometer CM 825 (Courage & Khazaka). Twenty women with an average age of 44.3 participated in the study; they were instructed not to apply any cosmetic or dermopharmaceutical composition other than those used in the study during its duration, or during the 24 hours prior to the beginning of the study.
• the skin hydration measurements were carried out 2 and 8 hours after the first application of the previous compositions as well as 20 days after the beginning of the study.
• Table 3 shows the average percentages of improvement to the skin's hydration of the placebo composition and the cosmetic composition from example 4 containing the exopolysaccharide of strain CNCM 1-4150.
• Placebo composition 12.1% 7.1 %
• Example 7 In vivo study of reduction of the skin's roughness.
• the measurements for this study were carried out in a bioclimatic room (24 ⁇ 2 °C; 50 ⁇ 10% relative humidity) in order to maintain a constant temperature and humidity during the measurements.
• the measurements of the skin's roughness were carried out through silicon replicas of the skin using adhesive discs (3M, 24x40) and a quick- setting synthetic polymer (SILFLO, Flexico Ltd).
• the silicon replicas of the skin were analyzed using image processing software (Quantilines, Monaderm) which enabled maximum value of roughness to be determined (depth of the wrinkle, referred to in the study as Rz).
• the anti-wrinkle effectiveness is shown by a decrease in the Rz value. Twenty women with an average age of 41 participated in the study; they were instructed not to apply any cosmetic or dermopharmaceutical composition other than those used in the study during its duration, or during the 24 hours prior to the beginning of the study.
• Table 4 shows the average percentages of percentage reduction of the maximum roughness (Rz) of the skin from the placebo composition and the cosmetic composition from example 3 containing the exopoiysaccharide of strain CNCM 1-4150. 1 * 2 hours — To e hours — To 20 days — To
• Example 8 Preparation of a cosmetic composition of the exopolysaccharide excreted by bacterial stain CNCM 1-4150 and Antarcticine ® .
• the cosmetic composition from this example was prepared by following the instructions for the preparation of the composition from example 3 with the ingredients and the quantities from table 5.
• phase D Antarcticine ® [INCI: Pseudoalteromonas Ferment Extract] was also added with sodium salicylate [INCI: Sodium Salicylate].
• Example 9 Preparation of a cosmetic composition of the exopolysaccharide excreted by bacterial strain CNCM 1-4150 and Serilesine ® .
• the cosmetic composition from this example was prepared by following the instructions for the preparation of the composition from example 3 with the ingredients and quantities of table 6.
• phase D Serilesine ® [INCI:Hexapeptide-10] was also added.
• the sample was passed through a microfluidifier for one cycle at an entrance pressure of 80 bar and 12500 psi on exit.
• the liposomes obtained were added to Luviquat ® HM 552 [INCI: Polyquaternium-16] in a liposome:cationic polymer ratio of 1.5:1 under light stirring.
• Example 11 Preparation of coacervation capsules of lipid nanoparticles containing a microemulsion of the exopolysaccharide excreted by bacterial strain CNCM 1-4150
• Phase B was slowly added to phase A under stirring.
• the mixture of phases A and B was added to the phase C ingredients, refined soybean oil IP Ph.
• Eur [INCI: Glycine Soja (Soybean) Oil], Arlacel 83V [INCI: Sorbitan Sesquioleate], and Massocare HD [INCI: Isohexadecane] (phase C) and a microemulsion was obtained.
• the mixture was homogenized under pressure in a microfluidifier for 3 cycles with an entrance pressure of 80 bar and pressure on exit of 15000 psi. Throughout the whole process the sample was maintained thermostated at 25°C using a water/glycol refrigeration circuit.

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