FR2813789A1 - Composition used for cosmetic treatment of skin comprises vitamin C and/or vitamin A and fucose component - Google Patents

Abstract

Composition comprises vitamin C, vitamin A and/or their derivatives and at least one fucose component comprising fucose, or a polysaccharide or oligosaccharide containing fucose and at least one excipient.

Description

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Title: Novel cosmetic or pharmaceutical composition comprising a combination of vitamin C and / or A with a fucose component, and use of this combination in cosmetics or pharmacy.

 The present invention relates to a new cosmetic or pharmaceutical composition comprising a combination of vitamin C and / or A with a fucose component, and the use of this combination, especially in products with topical application, for which an activity on the epithelial or connective tissue is sought, in particular anti-aging products, such as pharmaceutical or veterinary products, and more particularly, in cosmetics.

 Collagen, a major constituent of the dermis underwent, according to previous work (BRANCHET et al., Arch Gerontol Geriatr., 1991, 13: 1-14), a quantitative decrease with age. The regulation of its biosynthesis is therefore a very important step to consider in the fight against skin aging.

 Although it is well known that ascorbic acid (or vitamin C) and its salts (in particular sodium) have a stimulating effect on the biosynthesis of collagen, it was possible to demonstrate its cytotoxic effect on the other hand. concentration of about 0.01% by weight.

 The activation effect of collagen biosynthesis, which is very favorable, can be observed on electron microscopy slides, by a strong dilation of the vesicles of the rough endoplasmic reticulum, filled with neosynthesized proteins.

 The adverse cytotoxic effect, observable at millimolar concentrations of ascorbate (approximately 2.5 mM), is manifested by the detachment of cells from their substrate, the slowing down of their proliferation and cell viability, and then by cell death. .

 Furthermore, retinol (or vitamin A) and other retinoids such as retinyl palmitate are preferred compounds especially in the field of cosmetics for their favorable biological activities including the fight against skin aging. These activities revealed by topical use of vitamin A and its derivatives are presented for example in the article by Wade Cheng, PhD and Shirley De Petris, Vitamin A Complex, Skin Inc., March / April 1998.

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 However, the use of pure vitamin A or of one of its derivatives has the disadvantage of a toxicity such that the dosage must be limited or add components, in particular to minimize the discomfort of the irritation of the skin. resulting skin.

 In particular, it has been reported that retinol induces inhibition of cell proliferation on fibroblasts in conventional culture (Lacroix A., Anderson GDL, Lippman ME, Retinoids and cultured human fibroblasts, Exp Cell Res, 1980, 130: 339- 344, Harper RA, Burgoon T., Differential effects of retinoic acid on the growth of normal fibroblast-like cells in vitro from human, swine and rabbit skin, Cell Biol Int Rep, 1982, 6: 163-170, or Stumpenhausen G , Hein R., Kulozik M., Mauch C., Bryce GF, Oono T., Krieg Th., The influence of retinoids on fibroblast functions, in Saurat JH ed., Retinoids: 10 years On, 1991, pp.

 139-150 Karger, Basel). This is a true toxic effect of retinol on fibroblasts.

 Since vitamins C and A and the salts and derivatives thereof are vitamin components which are very often present in particular in cosmetics, in particular anti-aging cosmetics, it was therefore highly desirable to be able to overcome the abovementioned disadvantages in order to be able to increase the contents and thus the favorable effects of these components, while reducing their toxic effects.

 It has now been found, quite surprisingly and unexpectedly, that the combination of fucose or a polysaccharide or oligosaccharide containing fucose with vitamin C and / or vitamin A makes it possible to reduce significantly, by a true synergistic effect, the toxic effects of these vitamins.

 The present invention thus relates to a cosmetic or pharmaceutical composition characterized in that it comprises at least one vitamin component selected from the group consisting of vitamin C and its derivatives, vitamin A (or retinol) and its derivatives, and mixtures thereof, in combination with at least one fucose component selected from the group consisting of fucose, fucose-containing polysaccharides and oligosaccharides, and mixtures thereof, and at least one cosmetically or pharmaceutically acceptable excipient .

 By derivatives of vitamin C is meant in particular according to the invention salts, such as sodium ascorbate, and esters such as ascorbyl phosphate or ascorbyl palmitate.

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 The term retinol (or vitamin A) should be understood to include hydrogenated and non-hydrogenated isomers, such as 9-cis-retinol and didehydroretinol.

 By derivatives of vitamin A is meant in particular according to the invention retinoids other than retinol, in particular the esters obtained with retinol and acetic acid, propionic acid, palmitic acid or stearic acid. , and more particularly retinoic acid, retinaldehyde (or retinal) and retinyl palmitate.

 The term retinaldehyde should be understood to include the 4 trans, 13-cis, 11-cis and 9-cis stereoisomeric forms.

 The monosaccharide fucose is a deoxyhexose close to galactose which it has the steric conformation. However, the structure of fucose differs essentially from that of galactose in that the carbon-6 atom has a methyl group (-CH 3) and not a primary alcohol group (-CH 2 OH). This methyl group indeed confers on the fucose molecule an interesting partial hydrophobic character, compensated by the other hydroxyl groups on the other four carbon atoms present.

 The fucose monosaccharide that may be used in the composition according to the invention may be L-fucose, D-fucose or a mixture thereof. L-fucose and D-fucose can each be in the form alpha, beta or a mixture of these forms alpha and beta. These products are in particular marketed by SIGMA.

 Fucose appears early in phylogeny, polysaccharides of some algae and fungi contain relatively large amounts, alone or in combination with other chemical compounds. Fucose is also widespread in the plant kingdom and some bacteria synthesize too. Fucose can also appear in sulphated form as in fucans.

 Thus, the expression "polysaccharides and oligosaccharides containing fucose" means any polysaccharide or oligosaccharide comprising at least one fucose unit, including sulphated fucanated polysaccharides and oligosaccharides.

 Fucans are sulphated polysaccharides, forming part of the cellular walls of brown algae thalli (Pheophyceae). They are also present in certain marine animals, such as sea urchin and sea cucumber. The raw fucan, also called fucoidans, obtained by acid extraction from the cell walls of brown algae thalli, consists of a population heterogeneous molecules, which mainly comprises sulfated L-fucose polymers, mass

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 high average molar (100,000 to 800,000 g / mol). Fucan preparations of lower average molecular weight (less than 20 000 to 10 000 g / mol) were obtained, for example by mild hydrolysis of high molecular weight fucan as described in EP-0 403 377 or by radical depolymerization such as described in WO 97/08206, which may be mentioned in particular the product called Fucoidane, marketed by SIGMA.

 Among the fucose-containing polysaccharides and oligosaccharides that can be used in the composition according to the invention, other than fucans, mention may be made in particular of those sold by Solabia, such as the Fucogel 1000 polysaccharide.

The application WO 96/23057 describes the process for preparing such a polysaccharide, comprising the fucose-galactose-galacturonic acid repeat unit.

 We have also discovered a novel mixture of oligosaccharides containing fucose, hereinafter called Oligofucose mixture particularly suitable for the present invention.

 It is a mixture of non-sulphated oligosaccharides based on fucose, characterized in that it comprises oligosaccharides of less than 13 saccharide units comprising at least one non-reducing terminal fucose unit, and in that it can be obtained by a process comprising at least one step of degradation of a polysaccharide derived from a microorganism of the genus Klebsiella pneumoniae subsp. pneumoniae.

sulfate -O(S03)- . Les fucanes sont en particulier exclus de cette définition. According to the invention, the term "non-sulphated oligosaccharide based on fucose" means, in accordance with the general knowledge of a person skilled in the art, an oligosaccharide containing at least one saccharide unit fucose and having no group.

sulfate -O (SO 3) -. Fucans are in particular excluded from this definition.

 By "oligosaccharide comprising at least one non-reducing terminal fucose unit" is meant according to the invention, in agreement with the general knowledge of a person skilled in the art, an oligosaccharide containing at least one saccharide unit fucose in the terminal position of the chain. of the oligosaccharide, this fucose unit being connected to the following saccharide unit of the rest of the oligosaccharide by an acetal-type bond.

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 The numbers of saccharide units can be measured using techniques well known to those skilled in the art, in particular using the HPLC chromatography technique as described in the examples below.

 Preferably, the mixture of oligofucoses comprises, relative to the total weight of the mixture, at least 15% by weight, and more particularly preferably from 20 to 50% by weight, of oligosaccharides of less than 13 saccharide units comprising at least a fucose unit in a non-reducing end position.

 More particularly, the mixture of oligofucoses is characterized in that it further comprises, relative to the total weight of the mixture, from 25 to 45% by weight of oligosaccharides having from 13 to 24 saccharide units comprising at least one fucose unit in a non-reducing end position.

 More particularly, the Oligofucose Mixture is characterized in that it additionally comprises, relative to the total weight of the mixture, from 15 to 35% by weight of oligosaccharides of more than 54 saccharide units comprising at least one fucose unit. in a non-reducing end position.

 The Oligofucose Mixture being capable of being obtained by a process comprising at least one step of degradation of a polysaccharide derived from a microorganism of the genus Klebsiella pneumoniae subsp. pneumoniae, the oligosaccharides preferably comprise, at least in part, the fucose-galactose-galacturonic acid repeating unit.

 In particular, the oligofucose mixture is obtainable by the process comprising the steps of: a) growing the microorganism of the genus Klebsiella pneumoniae subsp. pneumoniae in an aqueous nutrient medium by aerobic fermentation of an assimilable carbohydrate source; b) recovering the polysaccharide formed from the fermentation must; c) subjecting the polysaccharide to moderate hydrolysis; d) subjecting the hydrolysis product of step c) to enzymatic hydrolysis; and e) deactivating the enzyme and then recovering the oligofucose mixture thus formed.

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 More particularly, these steps a) to e) can be described as follows.

Etapea)
The microorganism Klebsiella pneumoniae subsp. pneumoniae is the microorganism deposited in the National Collection of Cultures of Microorganisms under the number 1-1507, or a mutant of the latter. This microorganism is also described in detail in the application WO 96/23057.

 The aqueous nutrient medium can be any aqueous medium known to those skilled in the art, containing sources of carbon, nitrogen and inorganic salts such as those described in application WO 96/23057.

 The fermentation can be carried out at temperatures of the order of 25 to 35 C, at a pH of about 6.0 to 7.5, under conditions of aeration and stirring, for durations of the order from 2 to 4 days.

 The fermentation can be carried out in a conventional fermenter by inoculating sterilized nutrient medium, for example by heating to a temperature of about 120 C or by sterilizing filtration.

Step b)
At the end of the fermentation period, the fermentation broth is recovered from which a polysaccharide rich in fucose is isolated in the following manner.

 The fermentation broth is subjected to a heat treatment at a temperature in particular of between approximately 100 and approximately 130 ° C., preferably between approximately 115 and approximately 125 ° C. for a period of time in particular of approximately 30 minutes to approximately 2 hours and preferably of about 40 minutes to about 1 hour and a pH in particular between about 2 and about 5.5 and preferably between about 3 and about 5.5.

 The heat treatment product is filtered by conventional means such as a plate filter press.

 A clear, viscous and cell-free polysaccharide is thus obtained.

 A precipitation is then carried out in an alcohol solvent, preferably an alcohol solvent selected from ethanol, isopropanol and mixtures thereof. In particular, about 1 to about 3.0 volumes of solvent per 1 volume of polysaccharide and preferably about 1.3 to about 2.0 volumes of solvent per 1 volume of polysaccharide are used.

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 Vacuum drying is then carried out at a temperature in particular of between approximately 20 and approximately 60 [deg.] C. and preferably between approximately 30 and approximately 50 [deg.] C. until a powder is obtained.

Stages c) and d): polysaccharide
This is an essential combination of steps. It has surprisingly and unexpectedly been found that the combination of a moderate hydrolysis step, preferably by irradiation with gamma rays and / or by protolysis, with an enzymatic hydrolysis step makes it possible to advantageously obtain a yield of overall hydrolysis sufficient, similar to that of a conventional hydrolysis such as acid hydrolysis, but with the advantage of specific cuts of enzymatic hydrolysis. In particular, a conventional acidic hydrolysis does not make it possible to obtain the mixture of specific oligofucoses insofar as it leads to the obtaining of statistical cuts, prohibitive as to their randomness. The compounds resulting from a conventional acid hydrolysis have also proved to be biologically inactive.

Step c): Moderate polysaccharide
Moderate hydrolysis is carried out by a gamma ray treatment, a protolysis treatment or by these two successive treatments. Preferably, a gamma ray treatment is carried out successively and then a protolysis treatment.

 The treatment with gamma rays proves to cause a significant decrease in the viscosity due to a limited degradation, attributable to the action of free radicals. It can be achieved with irradiation means well known to those skilled in the art.

 This treatment with gamma rays, very penetrating rays, has the further advantage of sterilizing the polysaccharide by killing the present germs that could induce inflammation or even cause a granuloma. Thus, any bacterial attack is prevented without having to add antiseptic products to the medium which could undesirably interfere with the biological activities of the final product.

 The polysaccharide powder obtained in step b), optionally irradiated with gamma rays, can therefore also be subjected to a protolysis treatment. For this purpose, it is dissolved in aqueous solution in particular from 1 to 20% by weight and preferably from 2 to 10% by weight, relative to the total weight of the aqueous solution.

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 The aqueous solution is subjected to a heat treatment, that is to say to a heating at a temperature in particular between about 75 and about 120 C and preferably between about 90 and about 100 C, for a time in particular between 1 and 6 hours, in the presence of a proton generating resin such as those marketed and well known to those skilled in the art, that is to say a resin generating protons which result in cutting of glycosidic bonds with fixation of a molecule of water.

Step d): enzymatic hydrolysis
An acidic buffer such as citric acid buffer (4.15 g / kg) disodium hydrogen phosphate (about 10.75 g / kg) is introduced into the hydrolyzate obtained in step b). The temperature of the solution is adjusted to a temperature in particular of between approximately 25 and approximately 45 ° C. and preferably between approximately 30 and approximately 40 ° C.

 An enzyme preparation is introduced comprising at least one endofucosidase, preferably Fermizyme HCP as marketed by the company Gist Brocades, in a content of, in particular, between about 2 and about 20% by weight, and preferably between about 5 and about 15%. by weight, relative to the initial weight of polysaccharide powder used.

 The mixture thus obtained is kept under agitation for a time, in particular between about 8 and about 24 hours and preferably between about 10 and about 20 hours, at a temperature in particular between about 25 and about 45 ° C. and preferably between about 30 and about 30 ° C. and about 40 ° C., the pH being adjusted to 6 by the presence of the buffer mixture.

step e)
The hydrolysis product obtained after step d) is filtered by conventional means such as a filter press with plates.

 The solution collected is then heat-treated at a temperature in particular of between approximately 75 and approximately 120 [deg.] C. and preferably between approximately 90 and approximately 105 [deg.] C. for a time in particular of between approximately 10 and approximately 45 minutes and preferably between approximately 20 and approximately 35 minutes, in order to deactivate the enzyme and more particularly the fucosidase activity of this specific enzyme.

 It is allowed to cool to a temperature in particular of between approximately 20 and approximately 40 ° C.

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 During cooling, preservatives can be added to the solution.

 The whole is then filtered under sterile conditions and then the conditioning is carried out.

 The Oligofucose Mixture thus obtained can be characterized using techniques well known to those skilled in the art, in particular by HPLC, thin layer chromatography and other methods and chemical assays.

 It can thus be seen that the oligosaccharides of the oligofucose mixture are such that fucose is found mostly at the end of the chain, in a non-reducing end position.

 The favorable effects of the composition according to the invention are manifested at low concentrations of fucose component, preferably at a concentration of between about 0.001 and about 20% by weight, and more preferably between about 0.01 and about 10%. by weight, the concentration of vitamin component being preferably between about 0.001 and about 90% by weight, and more preferably between about 0.1 and about 10% by weight, based on the total weight of the composition.

 By appropriately selecting the respective concentrations of the vitamin component and the fucose component, it is thus possible to significantly reduce, or even eliminate, the toxic effect of the vitamin component and thus to use in the composition according to the invention vitamin component greater than or equal to those already existing products in commerce, without risk to the user.

 Preferably, the vitamin component: fucose component weight ratio is from about 800: 1 to about 1: 2, and more preferably from about 600: to about 1: 1.

 The cosmetically or pharmaceutically acceptable excipient may be any excipient among those known to those skilled in the art in order to obtain a composition according to the invention in the form of a cream, a lotion, a gel, an ointment, etc., optionally in the form of an emulsion with further components known to those skilled in the art, for improving, modifying or stabilizing the composition from a cosmetic or cosmetic point of view.

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 The term pharmaceutically acceptable excipient includes excipients suitable for veterinary use of the composition according to the invention.

 The composition according to the invention may in particular contain other additives and formulation aids, such as antioxidants to combat free radicals. It may be mentioned in particular pure vitamin E or di-alpha-tocopherol and its derivatives, and 2,6-di-tert-buthyl-p-cresol (BHT).

 According to a particular embodiment of the present invention, the composition further comprises a vector, such as microspheres containing in particular the vitamin component, for example the Talaspheres described in US-5,395,620 or in our patent application PI 9706994. -7.

 The composition according to the invention may thus for example comprise a plurality of dispersed microspheres, comprising a first vitamin C component in a first group of microspheres and a second vitamin A component in a second group of microspheres, the fucose component being outer microspheres, in the rest of the composition. Of course, a variant of this embodiment may consist in that the vitamin C and / or A component is included in a single group of microspheres.

 Advantageously, the composition according to the invention may in particular also comprise at least one cosmetically or pharmaceutically acceptable additive, selected from the group consisting of structuring agents of the skin (such as squalane and sphigolipids), humectants (such as glycerin and hydroxy prosilan C), emollients (such as butylene glycol and ethyl lactate), silicones (such as cyclomethicone), sunscreens (such as Parsol 1789 and Eusolex 6300) emulsifiers (especially Carbopol 1342 combined with triethanolamine and soy lecithin), thickeners (especially xanthan gum), sequestering agents (especially EDTA), antioxidants (such as the BHT described above) ) fragrances, preservatives, and mixtures thereof.

 Of course, the operating conditions for preparing the composition according to the invention are part of the general knowledge of those skilled in the art.

 The subject of the present invention is also the use, in a cosmetic or pharmaceutical composition, of at least one vitamin component as defined above.

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 above in combination with at least one fucose component as defined above, to reduce the toxic effects of the vitamin component.

 Preferably, a vitamin component weight ratio is used: the fucose component is from about 800: 1 to about 1: and more preferably from about 600: to about 1: 1.

 Finally, the subject of the present invention is also a method for the cosmetic or pharmaceutical treatment of the skin, characterized in that a cosmetic or pharmaceutical composition as defined above is applied to the skin.

 In particular, the subject of the present invention is a method for the cosmetic treatment of the skin, characterized in that a cosmetic composition as defined above is applied to the skin.

 The following examples illustrate a real synergy between fucose, fucose oligosaccharides with retinol and ascorbate and justify their association especially in a "anti-aging" cosmetological preparation.

 The following examples, however, are intended only to illustrate the present invention and should in no way be interpreted as limiting its scope.

 Figure 1 is a histogram showing the results presented in Example 4.b.l, in terms of percent efficiency of fucose and Oligofucose Mix-1 for stimulation of collagen synthesis by fibroblasts.

 FIG. 2 is a histogram showing the results presented in example 4.b.2, in terms of the percentage efficiency of fucose and Oligofucose Mix-1 for the stimulation of collagen biosynthesis by fibroblasts in the presence sodium ascorbate.

 Figure 3 is a histogram showing the results presented in Example 4.b.3, in terms of percent efficiency of fucose and Oligofucose Mix-1 for the stimulation of collagen synthesis in the presence of retinol.

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Example 1 Preparation of a Mixture of Oligofucoses a) Fermentation
The microorganism Klebsiella pneumoniae subsp. pneumoniae is the microorganism deposited in the National Collection of Cultures of Microorganisms under number 1-1507. The nutrient medium and other conditions of fermentation are as follows.

Preparation of inoculums Culture medium: Neosorb 70-07 (sorbitol content: 70% MS; sold by ROQUETTE FRERES, Lille / France): 17.90 g / 1 (12.5 g / 1 of sorbitol) Peptone Biokar 104003 (Protein hydrolyzate, sold by SOLABIA-BIOKAR, Pantin, France): 4.50 g / 1 yeast extract: 0.05 g / l KH 2 PO 4: 1.50 g / l K 2 HPO 4: 4.50 g / l MGSO 4, 7H20 0.20 g / l Pluronic # PE 61000 (antifoaming agent, sold by BASF, D-6700 Ludwigshafen, Germany): 0.50 g / 1 Solution dissolved in water.

Culture condition: Sterilization at 121 C for 30 minutes Culture temperature: 30 C Inoculation rate: 5 to 10% Aeration: 1 VVM unregulated PH (pH around 7.00) Culture time: 24 hours Medium of production Culture medium: Neosorb # 70-07: 54.00 g / 1 (ie 38 g / I sorbitol) Peptone Biokar #b 104003: 4.50 g / l Yeast extract: 0.05 g / l KH2PO 4: 1.50 g / 1

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 MgSO 4, 7H 2 O: 0.20 g / l Pluronic # PE 61000: 0.50 g / l Dissolution in water.

Culture conditions (Chemap fermenter with a useful volume of 350 liters): Sterilization at 120 ° C. for 45 minutes Culture temperature: 30 ° C Inoculation rate: about 5% Agitation: 300 rpm (Rushton type stirrer) Aeration: 1 VVM PH regulated at 7.0 with 7 N NaOH Pressure: 100 to 200 mbar Crop time: 60 to 65 hours Average values obtained in production: Viscosity at the end of the cycle: 40000 Mpa. s (Brookfield DV-II Viscometers + LV model, mobile SP 31, chamber SC4-34 / 13R, 30 C) Concentration of the polysaccharide produced in the medium, calculated in L-fucose: 2 g / l (Dische and Shettles method) Sorbitol consumed:> 35 g / 1 (in sorbitol) 20% NaOH in weight consumed: 15 liters / m3 Start of pH regulation: 16 to 17 hours after fermentor inoculation Final dry extract of fermentation medium: # 20 g / lb ) Recovery of formed polysacharide
The fermentation must is subjected to a heat treatment at a temperature of 120 ° C. for 45 minutes and at a pH of 5.5. The product of the heat treatment is filtered using a filter press with Seitz type plates. A clear, viscous and cell-free polysaccharide is thus obtained. Precipitation is then carried out in a 1.5 volume of ethanol per 1 volume of polysaccharide. Drying is then carried out under vacuum at a temperature of 25 ° C. until a powder is obtained. In view of the microorganism used, this polysaccharide is composed of fucose-galactose-galacturonic acid trisaccharide repeating units and thus has the following structure:

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c) Hydrolyse modérée du polysaccharide
La poudre de polysaccharide est mise en solution aqueuse à raison de 5 % en poids, par rapport au poids total de la solution aqueuse. La solution aqueuse est soumise à un traitement thermique, c'est à dire à un chauffage à 100 C, pendant 3 heures, en présence d'une résine génératrice de protons. d) Hydrolyse enzymatique
On introduit le mélange tampon acide citrique (4,15 g/kg)-hydrogénophosphate disodique (environ 10,75 g/kg) dans l'hydrolysat. On règle la température de la solution à 37 C. On introduit la préparation enzymatique Fermizyme HCP telle que commercialisée par la société Gist Brocades, selon une teneur de 10 % en poids, par rapport au poids initial de poudre de polysaccharide utilisée, c'est à dire 0,05% en poids par rapport à la masse totale de la solution aqueuse après remise en eau de la poudre comme décrit ci-dessus pour l'hydrolyse modérée par protolyse.

c) Moderate hydrolysis of the polysaccharide
The polysaccharide powder is dissolved in aqueous solution at a rate of 5% by weight, relative to the total weight of the aqueous solution. The aqueous solution is subjected to a heat treatment, that is to say to a heating at 100 C, for 3 hours, in the presence of a proton generating resin. d) Enzymatic hydrolysis
The citric acid (4.15 g / kg) disodium hydrogen phosphate (about 10.75 g / kg) buffer mixture is introduced into the hydrolyzate. The temperature of the solution is adjusted to 37 ° C. The fermizyme HCP enzyme preparation is introduced, as sold by the company Gist Brocades, at a content of 10% by weight, relative to the initial weight of polysaccharide powder used. ie 0.05% by weight based on the total mass of the aqueous solution after re-watering the powder as described above for moderate hydrolysis by protolysis.

The mixture thus obtained is stirred for 15 hours at a temperature of 37 ° C., the pH being adjusted to 6 by the presence of the buffer mixture. e) Deactivation of the enzyme and recovery of the oligosaccharide mixture
The hydrolysis product is filtered with a press filter with Seitz type plates.

The collected solution is then heat-treated at 100 ° C. for 30 minutes to deactivate the enzyme. It is allowed to cool to a temperature of 25 ° C. During cooling, the preservatives phenoxy ethanol (1% by weight) and phenonip (0.3% by weight) are added to the solution. The whole is then filtered under sterile conditions.

 The oligosaccharide mixture thus obtained is referred to as "Oligofucose Mixture-1".

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Exemple 2 : caractérisation HPLC du Mélange d'oligofucoses-1
Le fractionnement du Mélange d'oligofucoses-1 obtenu à l'exemple 1 a été réalisé dans le dessin de déterminer la proportion d'oligo- et polysaccharides dans sa composition. a) Fractionnement par chromatographie d'exclusion préparative sur la colonne " Xk 50/60 Superdex 75 prepgrade "
Nous avons passé 50ml du Mélange d'oligofucoses-1 concentré à 50mg/ml, sur une colonne préparative " XK 50/60 Superdex 75 prepgrade " (chromatographie d'exclusion) et nous avons collecté 95 fractions, passées ensuite en HPLC .

Example 2: HPLC Characterization of Oligofucose Mixture-1
Fractionation of the Oligofucose Mixture-1 obtained in Example 1 was carried out in the drawing to determine the proportion of oligo- and polysaccharides in its composition. a) Fractionation by preparative exclusion chromatography on the column "Xk 50/60 Superdex 75 prepgrade"
We passed 50ml of concentrated Oligofucose-1 Blend at 50mg / ml, on a preparative column "XK 50/60 Superdex 75 prepgrade" (exclusion chromatography) and we collected 95 fractions, then passed in HPLC.

Table 1: Technical information about the preparative column
XK 50/60 Superdex 75 prepgrade

<Tb>
<tb> Filling <SEP> Superdex <SEP> 75 <SEP> prepgrade <SEP> (34 <SEP> m)
<tb> Size <SEP> of <SEP><SEP> column <SEP> Height <SEP>: <SEP> cm
<tb> Diameter <SEP>: <SEP> 60 <SEP> mm
<tb> Type <SEP> of <SEP> column <SEP> XK <SEP> 50/60 <SEP>
<tb> Usable <SEP> Interval <SEP> of <SEP> 5 <SEP> x <SEP> 102 <SEP> Da <SEP> - <SEP> 3 <SEP> x <SEP> 104 <SEP> Da
<tb> splitting
<tb> Sample <SEP> Injection <SEP>: <SEP> 50 <SEP> ml <SEP> of <SEP><SEP> Blend of Oligofucoses-1 <SEP> to <SEP>
<tb> concentration <SEP> of <SEP> 50 <SEP> mg / ml. <SEP> (at <SEP> total <SEP>: <SEP> = <SEP>
<tb> 2.50 <SEP> g)
<tb> Elution rate <SEP><SEP> 1 <SEP> ml / <SEP> minute
<tb> Number <SEP> of <SEP> fractions <SEP> 95 <SEP> fractions <SEP> of <SEP> 12.5 <SEP> ml <SEP> each
<tb> collected
<tb> Mobile <SEP> Phase <SEP> PBS
<Tb>

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After fractionation of the Oligofucose-1 mixture on the "XK 50/60 Superdex 75 prepgrade" column, 95 fractions are collected, 45 of which contain osides b) Characterization of the fractions obtained by HPLC (exclusion chromatography) ultrahydrogel 120 and ultrahydrogel columns 250
The objective of this second part of our study was to pass on an HPLC exclusion column (Utrahydrogel columns 120 and 250) every 95 fractions of Oligofucose Mix-1 to analyze the molecular weights and the concentration of these components. fractions.

 We worked for this study with a Waters HPLC system, the description of which follows.

Table 2: Technical characteristics of the HPLC chromatography system used

<Tb>
<tb> Device <SEP> HPLC <SEP> Waters <SEP> 600
<tb> Columns <SEP> Ultrahydrogel <SEP> 120 <SEP> (size <SEP> of <SEP> pores <SEP>: <SEP> 120 <SEP>) <SEP> and <SEP> Ultrahydrogel <SEP> 250
<tb> (size <SEP> of <SEP> pores <SEP>: <SEP> 250 <SEP>) <SEP> of <SEP> Waters. <SEP> Size <SEP>: <SEP> 7.8 <SEP> mm <SEP> x <SEP> 300 <SEP> mm,
<tb> containing <SEP> the <SEP> gel <SEP> of <SEP> polymethacrylate <SEP> hydroxylated
<tb> Samples <SEP> injected <SEP> 20 <SEP> l <SEP> by <SEP> automatic <SEP> injector
<tb> Elution <SEP> 0.10 <SEP> M <SEP> NaN03 <SEP>
<tb> Time <SEP> of <SEP> elution <SEP> 50 <SEP> minutes / <SEP> samples
<tb> Speed <SEP> of <SEP> elution <SEP> 0.5 <SEP> ml / <SEP> minute.
<Tb>

Detection <SEP> By <SEP> measurement <SEP> of <SEP><SEP> index <SEP> refraction <SEP> with <SEP> a <SEP> refractometer
<tb> Waters <SEP> 410.
<Tb>

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Table 3: Molecular Weight Standards of Polyethylene Glycol (Fluka) Used for HPLC Exclusion Chromatography

<Tb>
<tb> Weight <SEP> molecular <SEP> SEP <SEP> TIME <SEP> (MINUTES)
<tb> 400 <SEP> 37,892
<tb> 600 <SEP> 36,026
<tb> 1000 <SEP> 33,940
<tb> 2000 <SEP> 31,154
<tb> 4000 <SEP> 28,896
<tb> 6000 <SEP> 27,868
<tb> 8000 <SEP> 26.946
<tb> 12000 <SEP> 26,192
<tb> 20000 <SEP> 25.308
<tb> 35000 <SEP> 24,216
<Tb>
c) Results
The molecular weights of the components of the fractions studied were calculated using the following equation, obtained with the Fluka molecular weight standards described in Table 3:
Molecular weight of the components: = 55290000 * 10 ^ (- 0.13942 * x) R '= 0.982 x = elution time (minutes)

<Desc / Clms Page number 18>

The first fraction containing components of Oligofucose Mix-1 is fraction No. 44 and the last fraction is No. 89, which means that even the lowest molecular weight component is removed after 89 fractions collected (after elution of 89 x 12.5 ml = 1112.5 ml). The collected fractions contain 184Da mono-, oligo- and polysaccharides (mixture of monosaccharides) up to about 21 kDa. This latter fraction therefore contains polysaccharides composed on average of 117 monosaccharide units or 39 trisaccharide units.

 The majority of the fractions, except fractions n 77, 78, 79, 81, 82, 83, 84, 85 and 86, contain a single saccharide peak (separation limited by the sensitivity of the separation method used).

 The approximate concentration of the components of the different fractions could be determined using a fucose standard range at increasing concentrations. This kind of "mono-compositional" standard range could be used thanks to the detection system (measurement of the refractive index with a refractometer).

According to these results, a solution of 1 g / ml of fucose gives on average a 29409 surface peak (arbitrary units of the system). By knowing the surfaces of the peaks analyzed, we were able to calculate their apparent concentrations.

 The results obtained show that the mixture of oligofucoses-1 contains about 26% small osids (up to 2 Kda, about 4 trisaccharide units), about 36% oligosaccharides (up to 4 Kda, 8 trisaccharide units) and about 23% % of polysaccharides of molecular weight greater than 10 kDa (18 trisaccharide units).

In view of the microorganism and the specific enzyme (endo-fucosidase) used for the preparation of Oligofucose Mix-1, it appears that the oligosaccharides of the Oligofucose-1 mixture comprise a non-reducing terminal fucose unit.

EXAMPLE 3 Action of the Association of Olixiducose Mixture with Sodium Ascorbate and / or Retinol on Human Skin Fibroblasts
The mixture of oligofucoses-1 as prepared in Example 1 is tested at concentrations of 1 μg / ml and 10 μg / ml, alone and in the simultaneous presence of 375 μg / ml of sodium ascorbate and / or 20 μg / ml of retinol.

<Desc / Clms Page number 19>

a) Methodology al) Study of cell proliferation
The human skin fibroblasts used in this study come from a skin sample of a 20-year-old woman (28th passage). The cells were cultured in 12-well plates, in DMEM culture medium with 10% fetal calf serum (FCS), 1% antibiotics and antifungals (FFP), and 1 Ci / ml. [3H] -thymidine (ICN) for 72 hours in the presence of the test products in the final concentrations of 1 g / ml and 10 g / ml.

After 72 hours of incubation (5% (v / v) CO2, 95% (v / v) air) at 37 ° C. in the presence of the tested products, the cells were washed four times with PBS and then the cell layer. detached by 0.05% trypsin. Three ml of scintillation fluid is then added per sample, and the radioactivity incorporated into the cells is read in a scintillation counter. at. 2) Synergy with retinol
We incubated the cells with 20 g / ml retinol (69.72 μM). Retinol induces, in the absence of oligosaccharides, a decrease in cell proliferation of approximately 45% (Table 2 below). Two concentrations were tested: 1 g / ml and 10 μg / ml. at. 3) Study of synergy with Na ascorbate
The cells were incubated with 375 g / ml Na ascorbate (1.875mM). Vitamin C induces, in the absence of fucose or Oligofucose Mix-1, a decrease in cell proliferation of approximately 60% (Table 3 below). Two concentrations were tested: 1 g / ml and 10 g / ml. at. 4) Synergy with Na Ascorbate and Retinol
Cells were incubated simultaneously with 375 g / ml Na ascorbate and 20 μg / ml retinol. This combination produces a strong inhibition of proliferation, partially lifted by the tested products.

<Desc / Clms Page number 20>

b) Results bl) Action on cell proliferation
The 72 hour incubation of the fibroblasts with Oligofucose Mix-1a stimulated cell proliferation compared to untreated cells (Table 1 below). b. 2) Synergy with retinol
The 72 hour incubation of the fibroblasts with 20 μg / ml retinol caused a 42.5% decrease in cell proliferation (Table 2 below) relative to the control without retinol.

 In the presence of 20 g / ml of retinol and different concentrations of the product tested, added simultaneously to the fibroblast culture media, cell proliferation is significantly higher than in the presence of 20ug / ml retinol alone. This is a protective effect demonstrating a synergy between Retinol and Oligofucose Mix-1.

de Na ont diminué l'incorporation de la [3 H]-thymidine, donc la prolifération cellulaire, d'environ 36 % par rapport au contrôle (tableau 3 ci-après). 1 μg / ml and 10 μg / ml of Oligofucose Mix-1 significantly increased cell proliferation (+ 24% and + 27%, respectively) compared to the control (20 g / ml retinol alone, Table 2 below). b. 3) Synergy with Na ascorbate
72 hours incubation of fibroblasts with 375 g / ml (1.875 mM) ascorbate

Na decreased the incorporation of [3H] -thymidine, thus cell proliferation, by about 36% compared to control (Table 3 below).

In the simultaneous presence of 375 g / ml of Na ascorbate and different concentrations of the test product, cell proliferation of fibroblasts increased significantly compared to ascorbate alone (Table 3 below). Oligofucose Mix-1 is effective at both 1 and 10 g / ml. b.4) Synergy with Na ascorbate and retinol
In the simultaneous presence of 375 μg / ml of Na ascorbate and 20 μg / ml of retinol, cell proliferation of fibroblasts decreased by about 88% compared with ascorbate alone (Table 4 below).

<Desc / Clms Page number 21>

TABLE 4: EFFECT OF THE DIFFERENT CONCENTRATIONS OF MIXTURE OF OLIGOFUCOSES-1 ON THE CELL PROLIFERATION OF HUMAN SKIN FIBROBLASTS (PROLIFERATION IN RELATION TO CONTROL)

<Tb>
<tb> Efficiency <SEP> by <SEP> Sign. <SEP> statistics
<tb> report <SEP> to <SEP> (p <SEP> by <SEP> report <SEP> to
<tb> Product <SEP> Concentration <SEP> control <SEP> control)
<tb> Control
<tb> Mixture <SEP> of oligofucoses-1 <SEP> 1 <SEP> g / ml <SEP> +4,4 <SEP> NS
<Tb>
<Tb>

Mixture <SEP> of oligofucoses-1 <SEP> 10 <SEP> g / ml <SEP> + 20.6 <SEP> 0.019
<Tb>
TABLE 5 Effect of the Different Concentrations of Oligofucose Mix-1 on the Cytotoxicity of 20 g / ml Retinol (Proliferation Versus Control and Compared with 20 g / ml Retinol)

<Tb>
<tb>% <SEP> vs. <SEP> Sign. <SEP> Sign.
<Tb>

Treatment <SEP> Concentration <SEP> 20 <SEP> g / ml <SEP> of <SEP>% <SEP> vs. <SEP> statistics <SEP> statistics
<tb> Retinol <SEP> Control <SEP> (p <SEP> vs. <SEP> Retinol) <SEP>
<tb> (Pvs <SEP> control)
<tb> Retinol <SEP> 20 <SEP> g / ml <SEP> -42.5 <SEP> 0.019
<Tb>
<tb><SEP> Mixture of Oligofucoses-1 <SEP> 1 <SEP> g / ml <SEP> + <SEP> 24.2 <SEP> -29.3 <SEP> 0.047 <SEP> 58
<tb> + <SEP> ascorbate <SEP> 375 <SEP> g / ml
<tb> *
<tb> Mixture <SEP> of Oligofucoses-1 <SEP> 10 <SEP> g / ml <SEP> + <SEP> 26.9 <SEP> -27.1 <SEP> 0.046 <SEP> 54
<tb> + <SEP> ascorbate <SEP> 375 <SEP> g / ml
<Tb>

TABLEAU 6 : Effet des différentes concentrations du Mélange d'olizofucoses-1 sur la cytotoxicité de 375 g/ml d'ascorbate de Na (prolifération par rapport au contrôle et par rapport au 375 g/ml d'ascorbate)

TABLE 6 Effect of the Different Concentrations of the Olizofucose Mix-1 on the Cytotoxicity of 375 g / ml Na Ascorbate (Proliferation versus Control and Compared to 375 g / ml Ascorbate)

<Tb>
<tb>% <SEP> vs. <SEP> Sign. <SEP> Sign.
<Tb>

Treatment <SEP> Concentration <SEP> 375 <SEP> g / ml <SEP>% <SEP> vs. <SEP> statistics <SEP> statistics
<tb> ascorbate <SEP> control <SEP> (pvs.
<tb> e <SEP> ascorbate) <SEP> (pvs.
<tb> control)
<tb> Ascorbate <SEP> of <SEP> Na <SEP> 375 <SEP> g / ml <SEP> -64.4 <SEP> 05
<tb> **
<tb> Mixture <SEP> of Oligofucoses-1 <SEP> 1 <SEP> g / ml <SEP> +96.9 <SEP> - <SEP> 30.0 <SEP> 0.003 <SEP> 51
<tb> + <SEP> ascorbate <SEP> 375 <SEP> g / ml
<Tb>
<tb> Mixture <SEP> of oligofucoses-1 <SEP> 10 <SEP> g / ml <SEP> + <SEP> 80.8 <SEP> - <SEP> 35.7 <SEP> 0.015 <SEP> 33
<Tb>

+ ascorbate 375 tg/ml 1 1 1 1 1

+ ascorbate 375 tg / ml 1 1 1 1 1

<Desc / Clms Page number 22>

TABLE 7 Effect of Different Concentrations of Oligofucose Mix-1 on the Cytotoxicity of 375 g / ml Na Ascorbate and 20 g / ml Retinol (Proliferation versus Control versus 375 g / ml ascorbate + 20 g / ml retinol)

<Tb>
<tb>% <SEP> vs. <SEP> Sign. <SEP> Sign.
<Tb>

Treatment <SEP> Concentration <SEP> ascorbate <SEP>% <SEP> vs. <SEP> statistics <SEP> statistics
<tb> + <SEP> control <SEP> (p <SEP> vs.
<tb> retinol <SEP> + retinol <SEP> (pvs.
<Tb>

+ ascorbate) <SEP> control)
<tb> Ascorbate <SEP> of <SEP> Na <SEP> 375 <SEP> g / ml <SEP> -64.4 <SEP> 05
<tb> Retinol <SEP> 20 <SEP> g / ml <SEP> -42.5 <SEP> 19
<tb> Ascorbate <SEP> of <SEP> Na <SEP> 375 <SEP> g / ml
<tb> + <SEP> Retinol <SEP> + <SEP> 20 <SEP> g / ml <SEP> - <SEP> 87.54 <SEP> 03
<tb> *
<tb> Mixture <SEP> of Oligofucoses-1 <SEP> 1 <SEP> g / ml <SEP> +95.2 <SEP> -75.7 <SEP> 0.010 <SEP> 04
<Tb>

+ ascorbate 375 jg/ml~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

+ ascorbate 375 μg / ml ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

<Tb>
<tb> **
<tb><SEP> Mixture of Oligofucoses-1 <SEP> 10 <SEP> g / ml <SEP> + 102.8-74.7 <SEP> 0.007 <SEP> 04
<Tb>

+ ascorbate 375 tg/ml 1 1 1 1 1

Exemple 4: effet du fucose et du Mélange d'oligofucoses-1 en présence d'ascorbate de sodium et/ou de rétinol
Nous avons étudié l'influence du fucose et du Mélange d'oligofucoses-1, tel que préparé à l'exemple 1, sur l'effet cytoxique de l'ascorbate de sodium ainsi que l'influence de ces deux composants fucose sur l'effet du rétinol. a) Méthodologie
Les fibroblastes de plastie mammaire d'une femme de 45 ans, au passage 14 ont été ensemencés dans les plaques de 12 puits à raison de 0,5. 105 cellules par puits. Les cellules sont mises en culture pendant 48 heures en présence du milieu de culture DMEM à 10% de serum de veau f#tal (SVF), à 1' étuve (5 % (v/v) CO2, 95% (v/v) air) à 37 C.

+ ascorbate 375 tg / ml 1 1 1 1 1

Example 4 Effect of Fucose and Oligofucose Mixture-1 in the Presence of Sodium Ascorbate and / or Retinol
We studied the influence of fucose and Oligofucose Mix-1, as prepared in Example 1, on the cytotoxic effect of sodium ascorbate and the influence of these two fucose components on the effect of retinol. a) Methodology
Mammoplasty fibroblasts of a 45-year-old woman at passage 14 were seeded in the 12-well plates at a rate of 0.5. 105 cells per well. The cells are cultured for 48 hours in the presence of DMEM culture medium containing 10% fetal calf serum (FCS), 1 oven (5% (v / v) CO2, 95% (v / v). ) air) at 37 C.

<Desc / Clms Page number 23>

 After rinsing with PBS, they are treated and re-cultured for 48 hours in the presence of test products and DMEM without FBS.

 The method is based on a specific colouration of collagen by Sirius red. The cells are fixed directly by Bouin's liquid (1 ml / well) for 1 hour, after thorough rinsing with PBS. The fixative is then aspirated and the plates are rinsed with running water by immersion for 15 minutes.

 Staining is done with stirring for 1 hour (1 ml / well) and the plates are then rinsed with 0.0N hydrochloric acid. The material is then dissolved in 200 l of 0.1 N sodium hydroxide prior to transfer to microtiter plates (Nunc). The optical density is measured at 550 nm against sodium hydroxide as white.

 Cells were counted on 4 wells of each plate, detaching the cells with 0.05% trypsin.

We studied the effect of fucose and Oligofucose Mix-1 (at 10 g / ml each) in the presence or absence of sodium ascorbate at 500 g / ml. Then in the presence and absence of retinol (10 g / ml). b) Results bl) Effects of Fucose and Oligofucose Mix-1
We found a decrease (about 19%) in the amount of collagen synthesized by fibroblasts in the presence of fucose. This decrease is much greater with the concentrations 1 g / ml and 10 g / ml than with 100 g / ml. Oligofucose Mixture-1 has no significant effect at 10 g / ml. These results are shown in Figure 1. b. 2) Effects of Fucose and Oligofucose Mix-1 in the Presence of Sodium Ascorbate
We have seen an increase in the amount of collagen synthesized by fibroblasts. Sodium ascorbate causes not only a disappearance of the inhibition expected by fucose but it even induces a stimulation of the

<Desc / Clms Page number 24>

 collagen biosynthesis. The same stimulatory effect of ascorbate is observed in the presence of Oligofucose Mix-1, but the observed stimulation is lower.

Sodium ascorbate at 500 g / ml only very weakly activates collagen synthesis compared to the effect of 50 μg / ml. By adding fucose, there is an increase in this stimulation, which is more modest with Oligofucose Mixture-1. These results are shown in Figure 2. b. 3) Effects of Fucose and Oligofucose Mix-1 in the Presence of Retinol
Retinol alone has an inhibitory effect on collagen synthesis. In the presence of fucose this inhibition is largely lifted, whereas it is not only lifted but there is moreover a slight stimulation in the presence of the Oligofucose-1 mixture.

These results are shown in Figure 3. c) Conclusion
Free fucose exerts a slight inhibition on collagen biosynthesis by cultured fibroblasts while Oligofucose Mix-1 does not exert this effect.

 In the presence of sodium ascorbate at 500 g / ml, the inhibition by fucose disappears and there is even the existence of stronger stimulation in the presence of fucose. In the presence of Oligofucose Mix-1, we also find this stimulation although a little more modest.

 Retinol alone at 10 μg / ml inhibits the biosynthesis of collagen. This inhibition is completely removed by the addition of Oligofucose Mixture-1.

 These results are shown in Figures 1 to 3.

<Desc / Clms Page number 25>

Example 5: anti-aging cream

<Tb>
<tb> Component <SEP>% <SEP> (in <SEP> weight)
<tb> Water <SEP> qsp <SEP> 100 <SEP>% <SEP>
<tb> Sodium <SEP> benzoate <SEP> 0.2
<tb> Disodium <SEP> EDTA <SEP> 0.08
<tb> Glycerin <SEP> 2.00
<tb> Butylene <SEP> Glycol <SEP> 4.00
<tb> Carbomer <SEP> ETD <SEP> 2020 <SEP> 0.20
<tb> Ceteareth-20 <SEP> 1.00
<tb> Mineral <SEP> Oil <SEP> 3.00
<tb> Squalane <SEP> 2.00
<tb> Octyl <SEP> Palmitate <SEP> 6,00
<tb> Shea <SEP> Butter <SEP> 2.50
<tb> Cetearyl <SEP> Alcohol <SEP> 1.00
<tb> Rosa <SEP> SEA <SEP> Rubiginosa <SEP> Seed <SEA> Oil <SEP> 0.20
<tb> Decyl <SEP> Oleate <SEP> 0.50
<Tb>

Octyl 1 Methox cinnamate 5,00

Octyl 1 Methox cinnamate 5.00

<Tb>
<tb> Butyl <SEP> Methoxy-dibenzoylmethane <SEP> 0.50
<tb> BHA <SEP> 0.01
<tb> Cyclomethicone <SEP> 5.00
<tb> Cyclomethicone <SEP>&<SEP> Dimethiconol <SEP> 2.00
<tb> Dimethicone <SEP> 2.00
<tb> Fragrance <SEP> (Crematest <SEP> Feno) <SEP> 0.09
<tb> Fragrance <SEP> (Chemoderm) <SEP> 0.09
<tb> Triethanolamine <SEP> 0.30
<Tb>

2-Bromo-2-Nitropropane- 1,3-Diol 0,02

2-Bromo-2-Nitropropane-1,3-Diol 0.02

<Tb>
<tb><SEP> Blend of Oligofucoses-1 <SEP> 0.50
<tb> Vitamin <SEP> A <SEP> 3.50
<tb> Vitamin <SEP> C <SEP> 1.00
<Tb>

<Desc / Clms Page number 26>

Example 6: anti-aging cream

<Tb>
<tb> Component <SEP>% <SEP> (in <SEP> weight)
<tb> Water <SEP> qsp <SEP> 100 <SEP>% <SEP>
<tb> Sodium <SEP> benzoate <SEP> 0.2
<tb> Disodium <SEP> EDTA <SEP> 0.08
<tb> Glycerin <SEP> 2.00
<tb> Butylene <SEP> Glycol <SEP> 4.00
<tb> Carbomer <SEP> ETD <SEP> 2020 <SEP> 0.20
<tb> Ceteareth-20 <SEP> 1.00
<tb> Ore <SEP> Oil <SEP> 3.00
<tb> Squalane <SEP> 2.00
<tb> Octyl <SEP> Palmitate <SEP> 6,00
<tb> Shea <SEP> Butter <SEP> 2.50
<tb> Cetearyl <SEP> Alcohol <SEP> 1.00
<tb> Rosa <SEP> SEA <SEP> Rubiginosa <SEP> Seed <SEA> Oil <SEP> 0.20
<tb> Decyl <SEP> Oleate <SEP> 0.50
<tb> BHA <SEP> 0.01
<tb> Cyclomethicone <SEP> 5.00
<tb> Cyclomethicone <SEP>&<SEP> Dimethiconol <SEP> 2.00
<tb> Dimethicone <SEP> 2.00
<tb> Fragrance <SEP> (Crematest <SEP> Feno) <SEP> 0.09
<tb> Fragrance <SEP> (Chemoderm) <SEP> 0.09
<tb> Triethanolamine <SEP> 0.30
<tb> 2-Bromo-2-Nitropropane-1,3-Diol <SEP> 0.02
<tb><SEP> Blend of Oligofucoses-1 <SEP> 0.50
<tb> Vitamin <SEP> A <SEP> 3.50
<tb> Vitamin <SEP> C <SEP> 1.00
<Tb>

Claims (22)

claims Cosmetic or pharmaceutical composition, characterized in that it comprises at least one vitamin component selected from the group consisting of vitamin C and its derivatives, vitamin A (or retinol) and its derivatives, and mixtures thereof, in combination with at least one fucose component selected from the group consisting of fucose, fucose-containing polysaccharides and oligosaccharides, and mixtures thereof, as well as at least one cosmetically or pharmaceutically acceptable excipient.  2. Composition according to claim 1, characterized in that the vitamin component is selected from the group consisting of ascorbic acid, salts and esters thereof, retinol, retinoids other than retinol, and mixtures of those -this.  3. Composition according to claim 1 or 2, characterized in that the vitamin component is selected from the group consisting of ascorbic acid, sodium ascorbate, ascorbyl phosphate, ascorbyl palmitate, retinol, retinoic acid. retinaldehyde, retinyl palmitate, and mixtures thereof.  4. Composition according to any one of the preceding claims, characterized in that the fucose component is selected from the group consisting of Lfucose, D-fucose, alpha, beta or a mixture of these alpha and beta forms. , fucans, polysaccharides and oligosaccharides comprising the fucose-galactose-galacturonic acid repeating unit, and mixtures thereof.  5. Composition according to any one of the preceding claims, characterized in that the fucose component is a mixture of non-sulphated oligosaccharides based on fucose, comprising oligosaccharides of less than 13 saccharide units comprising at least one terminal fucose unit. non-reducing, and in that it is obtainable by a process comprising at least one step of degradation of a polysaccharide derived from a microorganism of the genus Klebsiella pneumoniae subsp. pneumoniae.  6. Composition according to Claim 5, characterized in that the mixture of non-sulphated oligosaccharides based on fucose comprises, relative to the total weight of the mixture, at least 15% by weight of oligosaccharides of less than 13 units. <Desc / Clms Page number 28>  saccharides comprising at least one terminal non-reducing terminal fucose unit.  7. Composition according to Claim 5 or 6, characterized in that the mixture of non-sulphated oligosaccharides based on fucose comprises, relative to the total weight of the mixture, from 20 to 50% by weight of oligosaccharides of less than 13 units. saccharides comprising at least one terminal non-reducing terminal fucose unit.  8. Composition according to any one of claims 5 to 7, characterized in that the mixture of non-sulphated oligosaccharides based on fucose additionally comprises, relative to the total weight of the mixture, from 25 to 45% by weight of oligosaccharides having from 13 to 24 saccharide units comprising at least one non-reducing terminal fucose unit.  9. Composition according to any one of Claims 5 to 8, characterized in that the mixture of non-sulphated oligosaccharides based on fucose additionally comprises, relative to the total weight of the mixture, from 15 to 35% by weight of oligosaccharides of more than 54 saccharide units comprising at least one non-reducing terminal fucose unit.  10. Composition according to any one of claims 5 to 9, characterized in that the oligosaccharides of the mixture of non-sulphated oligosaccharides based on fucose comprise, at least in part, the fucose-galactose-galacturonic acid repeat unit.  11. Composition according to any one of claims 5 to 10, characterized in that the mixture of non-sulphated oligosaccharides based on fucose is obtainable by the process comprising the steps of: a) growing the microorganism of the genus Klebsiella pneumoniae subsp. pneumoniae in an aqueous nutrient medium by aerobic fermentation of an assimilable carbohydrate source; b) recovering the polysaccharide formed from the fermentation must; c) subjecting the polysaccharide to moderate hydrolysis; d) subjecting the hydrolysis product of step c) to enzymatic hydrolysis; and e) deactivating the enzyme and then recovering the oligosaccharide mixture thus formed. <Desc / Clms Page number 29>  12. Composition according to any one of claims 5 to 11, characterized in that the microorganism Klebsiella pneumoniae subsp. pneumoniae is the microorganism deposited in the National Collection of Cultures of Microorganisms under the number 1-1507, or a mutant of the latter.  13. Composition according to claim 11 or 12, characterized in that moderate hydrolysis, for the preparation of the mixture of non-sulphated oligosaccharides based on fucose, is carried out by a treatment selected from the group consisting of gamma ray treatments, protolysis treatments and combinations of these treatments.  14. Composition according to any one of claims 11 to 13, characterized in that the enzymatic hydrolysis, for the preparation of the mixture of non-sulphated oligosaccharides based on fucose, is carried out with at least one endo-fucosidase 15. Mixture according to claim 14, characterized in that the endo-fucosidase is Fermizyme HCP.  16. A composition according to any one of the preceding claims, characterized in that the concentration of the fucose component is between about 0.001 and about 20% by weight, and that the concentration of vitamin component is between about 0.001 and about 90%. % by weight, relative to the total weight of the composition.  17. Composition according to any one of the preceding claims, characterized in that the vitamin component: fucose component weight ratio is between about 800: 1 and about 1: 2.  18. Composition according to any one of the preceding claims, characterized in that it further comprises a vector in the form of microspheres containing the vitamin component.  19. Composition according to any one of the preceding claims, characterized in that it further comprises at least one cosmetically or pharmaceutically acceptable additive, selected from the group consisting of structuring agents of the skin, humectants, emollients, silicones, sunscreens, emulsifiers, thickeners, sequestering agents, antioxidants, fragrances, preservatives, and mixtures thereof.  20. Use in a cosmetic or pharmaceutical composition of at least one vitamin component as defined in any one of claims 1 to <Desc / Clms Page number 30>  3, in combination with at least one fucose component as defined in any one of claims 1 and 4 to 15, for reducing the toxic effects of the vitamin component.  21. Use according to claim 20, characterized in that the vitamin component: fucose component weight ratio is between about 800: 1 and about 1: 2.  22. Cosmetic treatment method of the skin, characterized in that a cosmetic composition as defined in any one of Claims 1 to 19 is applied to the skin.