# Patent

## Title
Combination of monosaccharides with ascorbic acid and use thereof

## Abstract

Composition containing a combination of at least one monosaccharide chosen from mannose, rhamnose and a mixture thereof, and of at least one additional compound chosen from ascorbic acid, an analogue thereof and a mixture thereof. The present invention also relates to the use of such a composition for combating the signs of ageing, and also to a device containing the composition.

## Background
REFERENCE TO PRIOR APPLICATIONS
This application claims priority to U.S. provisional application Ser. No. 61/145,119, filed Jan. 16, 2009; and to French patent application 08 59152, filed Dec. 30, 2008, both incorporated herein by reference.


FIELD OF THE INVENTION
The present invention relates to a composition, especially a cosmetic and/or dermatological composition, comprising, in a physiologically acceptable medium, a combination of at least one monosaccharide selected from mannose, rhamnose and a mixture thereof, and of at least one additional compound selected from ascorbic acid, an analogue thereof and a mixture thereof. The present invention also relates to the use of such a composition, and to a device containing it.
Additional advantages and other features of the present invention will be set forth in part in the description that follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from the practice of the present invention. The advantages of the present invention may be realized and obtained as particularly pointed out in the appended claims. As will be realized, the present invention is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the present invention. The description is to be regarded as illustrative in nature, and not as restrictive.
BACKGROUND OF THE INVENTION
Human skin is made up mainly of two main layers, namely the dermis and the epidermis that superficially covers the dermis. The dermis provides the epidermis with a solid support. It is also its nourishing element. It is made up mainly of fibroblasts and an extracellular matrix composed mainly of collagen, elastin and a substance known as ground substance. These components are synthesized by the fibroblasts.
The cohesion between the epidermis and the dermis is provided by the dermo-epidermal junction. This is a complex region about 100 nm thick, which comprises the basal pole of the basal keratinocytes, the epidermal membrane and the sub-basal zone of the superficial dermis. From a structural viewpoint, hemidesmosomes, into which are inserted keratin filaments (hemidesmosome-tonofilament complex), are distributed on the plasma membrane of the basal keratinocytes. Facing these hemidesmosome-tonofilament complexes are anchoring filaments that cross the epidermal basal membrane. The anchoring filaments are attached to laminin V on the epidermal side. Finally, anchoring fibrils constitute the sub-basal network. These are curvilinear structures that arise from and end on the deep face of is the basal membrane and in which are engaged fibres of collagen I, III and V.
It has been shown that these anchoring fibrils, which are entirely visible by electron microscopy, are composed of type VII collagen (referred to as collagen VII hereinbelow). The collagen VII is synthesized by keratinocytes and fibroblasts, but more substantially by keratinocytes (Aumailley M., Rousselle P. Laminins of the dermoepidermal junction. Matrix Biology, 1999, 18: 19-28; Nievers M., Schaapveld R., Sonnenberg A. Biology and function of hemidesmosomes. Matrix Biology, 1999, 18: 5-17).
Collagens are the major proteins of the extracellular matrices of the skin. To date, 20 types of collagen have been identified, and are noted from I to XX. The collagens predominantly present throughout the epidermis are collagens of the type I and III that form the extracellular matrix of the entire dermis (these collagens constitute 70-80% of the dry weight of the dermis). Moreover, collagens are not all synthesized by the same cell types: collagens of type I and III are essentially produced by the dermal fibroblasts, whereas type VII collagen is produced by two categories of cell, keratinocytes and fibroblasts. Regulation of their expression differs from one collagen to another, for example collagens I and VII are not regulated in the same way by certain cytokines; specifically, TNF-α and leukoregulin stimulate collagen VII and negatively regulate collagen I. Among the other types of collagens involved especially in ageing, mention may be made of collagens XII and VI. The collagen XII binds the fibrils of collagen Ito the other matrix compounds in the papillary dermis, and thus regulates the biomechanical properties of skin tissue: deformability and contraction of the collagen fibres by promoting sliding of the fibres relative to each other. The collagen VI facilitates, like proteoglycans, the three-dimensional arrangement of the collagen fibres. The special feature of collagen VI is its multiplicity of actions. Specifically, it binds with a large number of cells via receptors of integrin type, and with many matrix molecules (collagen IV, fibronectin, biglycan, MAGP-1). Finally, all collagen molecules are variants of a common precursor, which is the a chain of procollagen.
The dermoepidermal junction is a structure that conditions the surface state of the skin. Thus, a dermoepidermal junction with intact anchoring structures is maintained folded, thus making it possible to increase the surface area of the contact zone between the dermis and the epidermis, to promote exchanges of diffusible factors, especially between these two tissues, to reinforce their cohesion and to improve the appearance of the epidermis. In cases where the anchoring structures are impaired, in particular due to a deficiency in the synthesis of collagen VII or tenascin and/or due to ageing, this causes flattening of the dermoepidermal junction. Fewer exchanges take place, the two tissues are less solidly connected, the epidermis folds, and, as the skin is less firm and less taut, wrinkles appear and the fragility of the skin with respect to mechanical attack is increased.
Tenascin is a major constituent of the dermoepidermal junction. It is a an extracellular matrix glycoprotein, also known as tenascin-C. Its essential function lies in epithelium-mesenchyme interactions, especially during embryogenesis. In the skin, tenascin is found at the sub-epidermal level in the papillary dermis, but also around vessels and appendices. Its expression is greatly increased in situations of hyperproliferation, for instance psoriasis and tumours, but also in cicatrization. Tenascin is produced in the skin by two major cell types, the keratinocytes and the fibroblasts. One of its functions lies in cell adhesion. Specifically, the strong upregulation of tenascin in the migrating keratinocytes during cicatrization strongly suggests an essential role of adhesion of the keratinocytes to connective tissue, ensuring good dermoepidermal cohesion (Crossin K L. Tenascin: a multifunctional extracellular matrix protein with a restricted distribution in development and disease. J. Cell. Biochem. 1996, 61: 592-598; Latijnhouwers M., Bergers M., Ponec M., Dijkman H., Andriessen M., Schlkwijk J. Human epidermal to keratinocytes are a source of tenascin-C during wound healing. J. Invest. Dermatol., 1997, 108: 776-783; Steijlen P. M., Maessen E., Kresse H., Van Vlijmen I. M. J. J., Verstraeten A. A., Traupe H., Schalkwijk J. Expression of tenascin, biglycan and decorin in disorders of keratinization. Br. J. Dermatol., 1994, 130: 564-568).
With age, collagen becomes thinner and disorganised, skin cell renewal decreases, wrinkles appear on the surface of the skin, and the skin is duller and less firm. Cutaneous ageing is conditioned by genetic characteristics. Moreover, certain environmental factors such as smoking and above all exposure to sunlight accelerate it. The skin thus has a much more aged appearance on the areas exposed to sunlight, such as the back of the hands or the face. Thus, these other factors also have a negative impact on the natural collagen of the skin.
Consequently, given the important role of collagen in the integrity of the skin and in its resistance to external attacking factors of mechanical type, stimulation of the synthesis of these collagens, and in particular of procollagen I and collagens VI, VII and XII, appears to be an effective means for overcoming the signs of ageing of the skin.
To overcome the abovementioned drawbacks, to improve the appearance of the skin, to improve its mechanical properties and avoid pathologies associated with insufficiency or cell deficiency, deficiency in cell renewal or deficiency in certain compounds of the dermis or of the dermoepidermal junction, the inventors consider it to be important to develop products that are directed towards reinforcing or maintaining the role of the dermis as a support and nourishing element, the cohesion between the various layers of the skin, and more particularly the cohesion between the dermis and the epidermis, by increasing keratinocyte proliferation, stimulating fibroblasts proliferation and metabolism and stimulating collagen synthesis, in particular the synthesis of procollagen I and collagens VI, VII and XII, and increasing the synthesis of tenascin.
The epidermis, which covers the dermis and is in direct contact with the external environment, has the main role of protecting the body against the dehydration and external attack. Natural human epidermis is composed mainly of three types of cell, namely keratinocytes, which form the vast majority, melanocytes and Langerhans cells. Each of these cell types contributes, by virtue of its intrinsic functions, towards the essential role played in the body by the skin, especially the role of protecting the body against external attacking factors (the weather, ultraviolet rays, tobacco, etc.), which is known as the “barrier function”.
The epidermis is a keratinized, stratified pavement epithelium 90% made up of keratinocytes. The gradual differentiation of the cells of the basal membrane, which separates the dermis from the epidermis, towards the surface of the epidermis especially includes the differentiation of keratinocytes, which migrate towards the surface of the skin, where they desquamate.
Ageing of the epidermis is manifested mainly by a reduction in its thickness. Atrophy of the epidermis is the consequence of the slowing down of keratinocyte proliferation and of the accumulation of senescent keratinocytes. The horny layer becomes dull.
The cells constituting the epidermis are delimited by a lipid domain. In the course of differentiation, phospholipids, the role of which consists in producing the fluid structure of the cell membranes of the living layers of the epidermis, are gradually replaced by a mixture composed predominantly of fatty acids, cholesterol and ceramides (sphingolipids).
These lipids are organized in specific lamellar structures whose integrity depends not only on the quality of the fractions present, but also on their respective proportion. This lamellar structure of the lipids of the lipid domain of the epidermis is responsible for the fluidity and thus the suppleness of the skin. The lipids are also responsible for the “barrier” properties of the epidermis, particularly of the stratum corneum.
The epidermal lipids are mainly synthesized in living epidermis. They are made up mainly of phospholipids, sphingolipids, cholesterol, free fatty is acids, triglycerides, cholesterol esters and alkanes. The phospholipids are essential for the constitution of cell membranes. They play an important role in the mediation of extracellular signals and the formation of free aliphatic chains used for energy production. They constitute a reservoir of free fatty acids necessary for the constitution of the sphingolipids. The cholesterol plays a fundamental role in moisturization of the skin and in the “barrier” function of the epidermis. Free fatty acids play a major role in maintaining the lamellar structure of the lipids of the stratum corneum, and also in the constitution of cell membranes, where they are responsible for the membrane fluidity, but also for physiological processes such as the functioning of receptors or enzymatic activity.
Ceramides, which are other lipids playing a paramount role in the metabolism of the epidermis, are necessary for maintaining the multilamellar structure of the intercorneocytic lipids. They are also essential for the “barrier” function of the epidermis and for water exchanges, especially for overcoming age-related moisturization problems.
It is known practice from the literature to use agents such as ascorbic acid (vitamin C) or certain derivatives thereof to enable in particular an increase in the synthesis of ceramides (J. Invest. Dermatol. 109: 348-355, 1997; EP-1 145 706; EP-1 145 710). In vitro tests have also demonstrated, independently, that it is possible to increase the synthesis of tenascin and of collagen VII by adding vitamin C to the culture medium (EP-1 334 714).
Given the ever-increasing demand from users for improved solutions for combating the signs of biological or actinic ageing of the skin, there is a need to develop more efficient care methods.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows results obtained for keratinocyte proliferation under certain conditions, described in detail below.
is FIG. 2 shows results obtained for keratinocyte proliferation under certain conditions, described in detail below.
FIG. 3 shows the number of fibroblasts measured between an untreated control whole reconstructed skin, on the left, and a whole reconstructed skin treated with 5 mM of rhamnose, on the right.
FIG. 4 shows photographs of frozen sections of reconstructed.
FIG. 5 shows the amount of collagens VI and XII synthesized by dermal fibroblasts in the presence or absence of rhamnose and/or vitamin C.



DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
In this context, the present invention has shown, surprisingly, that a combination of at least one specific monosaccharide and of ascorbic acid or an analogue thereof makes it possible to obtain synergistic action both on the lipogenesis of the epidermis, and more particularly on the synthesis of the epidermal ceramides, but also on the synthesis of tenascin, collagen VI, XII and/or VII and/or on stimulation of the synthesis of procollagen I and on activation of keratinocytes and/or fibroblasts proliferation. Thus, this combination can efficiently combat the signs of ageing of the skin, in the dermis, the dermoepidermal junction and the epidermis. In particular, it is possible to counter age-related epidermal and/or dermal atrophy and/or to reinforce the cohesion between the dermis and the epidermis.
The term synergism and its deriviatives (synergistic, etc.) as used herein means a greater than additive effect.
The present application demonstrates the activation of keratinocyte and fibroblast proliferation and the stimulation of procollagen I synthesis by mannose or rhamnose. The use of compositions containing them thus makes it possible to counter the signs of ageing of the skin, and in particular age-related dermal and/or epidermal atrophy.
The use of these monosaccharides for the direct biological effects outlined above was hitherto unknown. Patent application WO 2007/128939 mentions, however, anti-ageing activity obtained via a biomechanical effect of a tensioning agent in combination with saccharide compounds, which make it possible to increase the expression of the skin cell mechanoreceptors. This increase in the expression of mechanoreceptors is described as increasing the sensitization of skin cells to respond to the effects of tensioning agents.
Patent application WO 2005/063194 describes a galenical base with very high tolerance especially comprising mannose or rhamnose. It is specified that such a galenical base can function only in combination with an active agent whose galenical base is only the vehicle. The dermal and/or cosmetic galenical bases disclosed are based essentially on the presence of the two polyols, namely mannitol and xylitol.
The present invention thus relates in one embodiment to a composition, especially a cosmetic and/or dermatological composition, comprising a combination of at least one monosaccharide chosen from mannose, rhamnose and a mixture thereof, with at least one additional compound chosen from ascorbic acid, an analogue thereof and a mixture thereof.
Use may be made of ascorbic acid, which is generally but not necessarily in L form since it is usually extracted from natural products, or analogues thereof.
On account of its chemical structure (α-keto lactone), which makes it very sensitive to certain environmental parameters such as light, the heat and aqueous media, it may be advantageous to use ascorbic acid in the form of a saccharide ester of ascorbic acid or a metal salt of phosphorylated ascorbic acid.
The saccharide esters of ascorbic acid that may be used in the invention are especially glycosyl, mannosyl, fructosyl, fucosyl, galactosyl, N-acetylglucosamine, N-acetylmuramic derivatives of ascorbic acid and mixtures thereof, and more especially ascorbyl-2 glucoside or 2-O-α-D glucopyranosyl of L-ascorbic acid or 6-O-D galactopyranosyl of L-ascorbic acid. The latter compounds and processes for preparing them are described in particular in documents EP-A-487 404, EP-A-425 066 and J-05 213 736.
For its part, the metal salt of phosphorylated ascorbic acid may be chosen from alkali metal ascorbyl phosphates, alkaline-earth metal ascorbyl phosphates and transition metal ascorbyl phosphates.
The ascorbic acid analogues are more particularly the salts thereof, such as, especially, sodium ascorbate, sodium or magnesium ascorbyl phosphate, the acetic ester of ascorbic acid, or sugars thereof, including saccharide esters and especially such as glycosyl ascorbic acid.
If necessary, stabilization of ascorbic acid towards oxidation may be obtained by combining it with maleic anhydride derivatives, as described in patent application EP 1 374 852, or with imidazole polymers, as described in patent application FR 2 832 630.
According to one preferred embodiment of the invention, the additional compound present in the composition according to the invention is not ascorbyl palmitate.
According to one particular embodiment of the invention, the composition according to the invention does not comprise a combination of xylitol and mannitol.
Mannose is a hexose that is the C2 epimer of glucose. Rhamnose (or 6-deoxymannose) formally constitutes the product of deoxygenation of mannose at C6. The monosaccharides according to the invention are in the D or L form of mannose and/or rhamnose or a mixture thereof, each form itself possibly being the alpha and/or beta anomer. The forms that are preferred according to the invention are D-mannose or L-rhamnose.
D-Mannose is present in plants, in particular certain fruit, including cranberries, or in hardwood (beech and birch). Rhamnose is found in nature in L form. D-Mannose and L-rhamnose are commercially available, for example from the company Danisco Sweeteners® and from the company Symrise.
In the present invention, the monosaccharide is preferably present as a monomer.
The present invention also relates in another embodiment to the use, especially the cosmetic or dermatological use, of a composition according to the invention as defined previously, administered orally, topically or via cutaneous injection, especially for caring for the skin and/or the scalp.
A composition in accordance with the invention as defined previously may especially be a cosmetic haircare composition, in particular for stimulating hair growth, for combating hair loss, for slowing down hair loss or for reinforcing the radiance of the hair.
Another embodiment of the present invention is a treatment method, in particular a cosmetic or therapeutic method, for reducing or preventing the signs of ageing of the skin or its integuments (hair, eyelashes, nails, etc.), by administration to an individual, preferably a human being, of an effective amount of at least one monosaccharide as defined previously in combination with an effective amount of at least one ascorbic acid analogue and/or of ascorbic acid itself.
The composition or combination according to the invention is more particularly intended to be applied to the areas of the body or face of individuals showing signs of ageing of the skin, for instance wrinkles.
The present invention also relates to the use, especially the cosmetic or dermatological use, of the composition or combination according to the invention for reducing and/or preventing the signs of ageing of the skin and/or its integuments.
According to one particular mode, the composition used in the context of the present invention does not comprise a combination of xylitol and mannitol.
The composition or combination according to the invention also makes it possible in particular to stimulate the regeneration of epidermal and dermal cells, in the skin or the integuments, in particular keratinocytes and fibroblasts, especially by increasing their proliferation. This therefore provides a method, especially a cosmetic method, which is especially effective for combating the signs of chronological ageing and/or photoageing.
The signs of photoageing correspond to internal degradations of the skin due to exposure to ultraviolet radiation (actinic ageing). The signs of chronological ageing correspond to internal degradations of the skin due to the intrinsic ageing of the individuals.
According to one preferred embodiment, the use according to the present invention is intended for improving the radiance of the complexion, for reducing and/or preventing the characteristics of wrinkles and/or fine lines, for improving and/or reducing the microrelief of the skin, and/or for improving the mechanical properties of the skin and/or for increasing the resistance of the skin to mechanical attack, such as rubbing, tensions or frictions and/or for promoting skin repair.
According to another aspect of the invention, the use of the composition or of the combination according to the invention makes it possible to improve the density of the skin, its firmness and/or the cohesion of its various compartments, in particular the cohesion of the dermis with the epidermis.
The present invention also relates to the use of the composition or combination according to the invention for preventively or curatively treating is wrinkles and/or fine lines, withered skin, lack of skin elasticity and/or tonicity, thinning of the dermis, degradation of collagen fibres, flaccid skin, thinned skin and/or any internal degradation of the skin caused by exposure to ultraviolet radiation.
The present invention also relates to the use of the composition or combination according to the invention for stimulating skin regeneration, in particular of the epidermis and/or the dermis, by means of better skin cell renewal, in particular of the epidermis and/or the dermis.
By acting on the dermoepidermal junction, keeping it folded, thus making it possible to increase the area of the contact zone between the dermis and the epidermis, to promote exchanges between these two tissues, to reinforce their cohesion and to improve the appearance of the epidermis, the compositions according to the invention make it possible to attenuate wrinkles and/or to make the skin firm.
The composition or combination according to the present invention has the effect of improving the lipid profile of the epidermis by modifying lipogenesis and reinforcing the integrity of the skin lipids, and thus of improving the barrier function of the skin and/or the suppleness of the skin.
One particularly preferred object of the present invention is the use of the composition or combination according to the invention for improving the lipid profile of the epidermis by modifying lipogenesis, and in particular causing an increase in the synthesis of ceramides.
Another preferred object of the present invention is the use of the composition or combination according to the invention for increasing keratinocyte and/or fibroblast proliferation, or for stimulating collagen synthesis, in particular the synthesis of procollagen I, collagen VII and collagen VI and/or XII.
Another particularly preferred object of the present invention is the use of the composition or combination according to the invention for increasing the synthesis of tenascin and/or collagen VII.
The amount of the active ingredients, selected from the monosaccharides and ascorbic acid and analogues as defined previously, to be used according to the invention depends on the desired cosmetic or therapeutic effect, and can thus vary within a wide range. A person skilled in the art can, on the basis of his general knowledge, readily determine the appropriate amounts.
Thus, and according to one preferred embodiment, the composition according to the invention comprises at least one monosaccharide as defined above in an amount of between 0.001% and 30% by weight relative to the total weight of the composition, in particular between 0.1% and 10% by weight and more particularly between 0.5% and 6% by weight relative to the total weight of the composition. According to one embodiment, and in particular when the additional compound is ascorbic acid, the composition according to the invention comprises at least one monosaccharide as defined above in an amount of between 0.4% and 30% by weight relative to the total weight of the composition, in particular between 0.4% and 10% by weight and more particularly between 0.4% and 6% by weight relative to the total weight of the composition.
According to one preferred embodiment, the composition according to the invention comprises vitamin C and/or at least one analogue thereof in an amount of between 0.001% and 30% by weight relative to the total weight of the composition, in particular between 0.1% and 10% by weight and more particularly between 0.5% and 6% by weight relative to the total weight of the composition.
The composition according to the invention is suitable for topical administration to the skin or its integuments, oral administration or cutaneous injection, in particular in the form of a sterile solution.
Preferably, the topical administrations according to the invention are in the form of a cream, a gel, a lotion, a milk, an oil, an ointment, a wax, a mousse, a paste, a serum, a pomade or a shampoo.
Preferably also, the oral administrations according to the invention are in the form of a gel capsule, a tablet or pills.
The monosaccharide according to the invention and the ascorbic acid or analogue thereof are more particularly present in the composition according to the invention as active agents (or active ingredients), in particular as sole active agents.
According to the invention, the terms “active agent” and “active ingredient” more specifically mean a compound which, when administered to an individual, in particular to a human being, plays a direct biological role on the body, in particular on the skin or its integuments, in particular without improving the biological or mechanical effect of another compound present in the composition according to the invention.
In general, the medium in which the active principles of the composition as defined previously are included is a physiologically acceptable medium, in particular a cosmetically or pharmaceutically acceptable medium, and may be anhydrous or aqueous. It may thus comprise at least one aqueous phase and/or at least one fatty phase.
The physiologically acceptable medium in which the compounds according to the invention may be employed, and also the constituents thereof, their amount, the galenical form of the composition, its mode of preparation and its mode of administration, may be chosen by a person skilled in the art on the basis of his general knowledge, as a function of the desired type of composition.
When the composition is a composition intended for topical administration, it may advantageously be in the form of aqueous or aqueous-alcoholic solutions, oil-in-water O/W) or water-in-oil (W/O) emulsions or multiple emulsions (triple: W/O/W or O/W/O), nanoemulsions, in particular O/W nanoemulsions, in which the size of the drops is less than 100 nm, aqueous gels, or dispersions of a fatty phase in an aqueous phase with the aid of spherules, these spherules possibly being polymer nanoparticles such as nanospheres and nanocapsules or lipid vesicles of ionic and/or nonionic type (liposomes, niosomes or oleosomes (as described in patent applications FR 2 709 666 and FR 2 725 369)).
These compositions are prepared according to the usual methods.
In addition, the compositions that may be used according to the invention may be more or less fluid and may have the appearance of a white or coloured cream, a pomade, a milk, a lotion, a serum, a paste or a mousse. They may optionally be applied to the skin in aerosol form. They may also be in solid form, for example in stick form.
For local application to the hair or the scalp, the composition may be in the form of aqueous, alcoholic or aqueous-alcoholic solutions; in the form of creams, gels, emulsions or mousses; in the form of aerosol compositions also comprising a propellant under pressure.
When the composition is in aqueous form, especially in the form of an aqueous dispersion, emulsion or solution, it may comprise an aqueous phase, which may comprise water, a floral water and/or a mineral water.
When the composition is an emulsion, the proportion of the fatty phase may range from about 5% to 80% by weight and preferably from about 2% to 50% by weight relative to the total weight of the composition. The oils, waxes, emulsifiers and co-emulsifiers used in the composition in emulsion form are chosen from those conventionally used in cosmetics. The emulsifier and the co-emulsifier are present in the composition in a proportion ranging from 0.3% to 30% by weight and preferably from 0.5% to 20% by weight relative to the total weight of the composition. The emulsion may also contain lipid vesicles.
When the composition is an oily solution or gel, the fatty phase may represent more than 90% of the total weight of the composition.
The oily phase may also comprise any common liposoluble or lipodispersible additive, as indicated hereinbelow.
It may especially comprise fatty substances such as waxes, pasty compounds, fatty alcohols or fatty acids. The oily phase contains at least one oil, more particularly at least one cosmetic oil. The term “oil” means a fatty substance that is liquid at room temperature (25° C.).
As oils that may be used in the composition of the invention, examples that may be mentioned include:

    
    
        hydrocarbon-based oils of animal origin, such as perhydrosqualene;
        hydrocarbon-based oils of plant origin, such as liquid triglycerides of fatty acids containing from 4 to 10 carbon atoms, for instance heptanoic or octanoic acid triglycerides, or alternatively, for example, sunflower oil, corn oil, soybean oil, marrow oil, grapeseed oil, sesameseed oil, hazelnut oil, apricot oil, macadamia oil, arara oil, coriander oil, castor oil, avocado oil, caprylic/capric acid triglycerides, for instance those sold by the company Stearineries Dubois or those sold under the names Miglyol 810, 812 and 818 by the company Dynamit Nobel, jojoba oil, shea butter oil and caprylyl glycol;
        synthetic esters and ethers, especially of fatty acids, for instance the oils of formulae R1COOR2 and R1OR2 in which R1 represents a fatty acid or a fatty alcohol residue containing from 8 to 29 carbon atoms and R2 represents a branched or unbranched hydrocarbon-based chain containing from 3 to 30 carbon atoms, for instance Purcellin oil, 2-octyldodecyl stearate, 2-octyldodecyl erucate, isostearyl isostearate; hydroxylated esters, for to instance isostearyl lactate, octyl hydroxystearate, octyldodecyl hydroxystearate, diisostearyl malate or triisocetyl citrate; fatty alcohol heptanoates, octanoates or decanoates; polyol esters, for instance propylene glycol dioctanoate, neopentyl glycol diheptanoate and diethylene glycol diisononanoate; and pentaerythritol esters, for instance pentaerythrityl tetraisostearate, or isopropyl lauroyl sarcosinate, sold especially under the trade name Eldew SL 205 by the company Ajinomoto;
        linear or branched hydrocarbons, of mineral or synthetic origin, such as volatile or non-volatile liquid paraffins, and derivatives thereof, petroleum jelly, polydecenes, isohexadecane, isododecane, hydrogenated polyisobutene such as Parleam oil, or the mixture of n-undecane (C11) and of n-tridecane (C13) sold under the reference Cetiol UT by the company Cognis;
        fluoro oils that are partially hydrocarbon-based and/or silicone-based, for instance those described in document JP-A-2 295 912;
        silicone oils, for instance volatile or non-volatile polymethylsiloxanes (PDMS) with a linear or cyclic silicone chain, which are liquid or pasty at room temperature, in particular volatile silicone oils, especially cyclopolydimethylsiloxanes (cyclomethicones) such as cyclohexadimethylsiloxane and cyclopentadimethylsiloxane; polydimethylsiloxanes comprising alkyl, alkoxy or phenyl groups, which are pendent or at the end of a silicone chain, these groups containing from 2 to 24 carbon atoms; phenyl silicones, for instance phenyl trimethicones, phenyl dimethicones, phenyltrimethylsiloxydiphenyl-siloxanes, diphenyl dimethicones, diphenylmethyldiphenyltrisiloxanes or 2-phenylethyl trimethylsiloxy silicates, and polymethylphenylsiloxanes; or
        mixtures thereof.
    
    


In the list of oils mentioned above, the term “hydrocarbon-based oil” means any oil mainly comprising carbon and hydrogen atoms, and possibly ester, ether, fluoro, carboxylic acid and/or alcohol groups.
The other fatty substances that may be present in the oily phase are, for example, fatty acids containing from 8 to 30 carbon atoms, for instance stearic acid, lauric acid, palmitic acid and oleic acid; waxes, for instance lanolin wax, beeswax, carnauba wax or candelilla wax, paraffin wax, lignite wax or microcrystalline waxes, ceresin or ozokerite, and synthetic waxes, for instance polyethylene waxes and Fischer-Tropsch waxes; silicone resins such as trifluoromethyl-C1-4-alkyl dimethicone and trifluoropropyl dimethicone; and silicone elastomers, for instance the products sold under the name KSG by the company Shin-Etsu, under the name Trefil, BY29 and EPSX by the company Dow Corning, or under the name Gransil by the company Grant Industries.
These fatty substances may be chosen in a varied manner by a person skilled in the art so as to prepare a composition having the desired properties, for example in terms of consistency or texture.
The emulsions generally contain at least one emulsifier chosen from amphoteric, anionic, cationic and nonionic emulsifiers, used alone or as a mixture, and optionally a co-emulsifier. The emulsifiers are chosen in an appropriate manner according to the emulsion to be obtained (W/O or O/W). The emulsifier and the co-emulsifier are generally present in the composition in a proportion ranging from 0.3% to 30% by weight and preferably from 0.5% to 20% by weight relative to the total weight of the composition.
For W/O emulsions, examples of emulsifiers that may be mentioned include dimethicone copolyols, such as the mixture of cyclomethicone and dimethicone copolyol sold under the trade name DC 5225 C by the company Dow Corning, and alkyl dimethicone copolyols such as the lauryl dimethicone copolyol sold under the name Dow Corning 5200 Formulation Aid by the company Dow Corning, and the cetyl dimethicone copolyol sold under the name Abil EM 90® by the company Goldschmidt. A crosslinked elastomeric solid organopolysiloxane comprising at least one oxyalkylene group, such as those obtained according to the procedure of Examples 3, 4 and 8 of patent US-A-5 412 004 and of the examples of patent US-A-5 811 487, especially the product of Example 3 (synthesis example) of patent US-A-5 412 004, such as the product sold under the reference KSG 21 by the company Shin-Etsu, may also be used as surfactants for W/O emulsions.
For O/W emulsions, examples of emulsifiers that may be mentioned include nonionic emulsifiers such as oxyalkylenated (more particularly polyoxyethylenated) fatty acid esters of glycerol; oxyalkylenated fatty acid esters of sorbitan; oxyalkylenated (oxyethylenated and/or oxypropylenated) fatty acid esters; oxyalkylenated (oxyethylenated and/or oxypropylenated) fatty alcohol ethers; sugar esters such as sucrose stearate; and mixtures thereof, such as the mixture of glyceryl stearate and PEG-40 stearate.
These compositions may also be O/W emulsions stabilized with particles, for instance the polymer particles described in patent FR 2 760 641, zo or crosslinked or non-crosslinked amphiphilic polymers, as described in patent applications FR 2 853 543 and FR 2 819 175.
In a known manner, the cosmetic composition may also contain adjuvants that are common in cosmetics, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preserving agents, antioxidants, solvents, fragrances, fillers, odour absorbers and dyestuffs. The amounts of these various adjuvants are those conventionally used in the cosmetics field, and range, for example, from about 0.01% to 10% of the total weight of the composition. Depending on their nature, these adjuvants may be introduced into the fatty phase, into the aqueous phase and/or into lipid spherules.
As solvents that may be used in the invention, mention may be made of lower alcohols, for instance ethanol, isopropanol, dipropylene glycol, butylene glycol and propylene glycol.
As hydrophilic gelling agents that may be used in the invention, non-limiting examples that may be mentioned include carboxyvinyl polymers (Carbomer®), acrylic copolymers such as acrylate/alkylacrylate copolymers, polyacrylamides, polysaccharides such as hydroxypropylcellulose, natural gums and clays, and lipophilic gelling agents that may be mentioned include modified clays such as bentones, metal salts of fatty acids, for instance aluminium stearates, hydrophobic silica, ethylcellulose and polyethylene.
When the monosaccharide is administered orally, the composition containing it may advantageously be in the form of a gel capsule, a tablet or pills. When the monosaccharide is administered via cutaneous injection, the composition containing it may be in particular in the form of a sterile solution.
The compositions of the invention may contain other hydrophilic or lipophilic active agents. These active agents are chosen especially from antioxidants, dermo-relaxing or dermo-decontracting agents, anti-ageing agents, anti-glycation agents, agents for stimulating the synthesis of dermal or epidermal macromolecules and/or for preventing their degradation, agents for stimulating fibroblast or keratinocyte proliferation and/or keratinocyte differentiation, agents for promoting maturation of the horny envelope, NO-synthase inhibitors, and agents for stimulating the energy metabolism of cells. Lists of these active agents are given hereinbelow for illustrative purposes, and should not in any way the considered as limiting.
Anti-ageing Agents:
Among the active agents that are known for combating the signs of ageing, especially ageing of the skin, mention may be made especially of:
vitamin B3, coenzyme Q10 (or ubiquinone), vitamin B9, vitamin E, vitamin E derivatives, such as the phosphate derivative, for instance TPNA® sold by the company Showa Denko, resveratrol or derivatives thereof, for instance Resveratrate® sold by the company Estee Lauder, retinol or derivatives thereof, and a mixture thereof.
Anti-glycation Agents:
The term “anti-glycation agent” means a compound that prevents and/or reduces the glycation of skin proteins, in particular dermal proteins such as collagen.
Anti-glycation agents that may especially be mentioned include extracts of plants of the Ericacea family, such as an extract of blueberry (Vaccinium angustifolium or Vaccinium myrtillus), for example the product sold under the name Blueberry Herbasol Extract PG by the company Cosmetochem, ergothioneine and derivatives thereof, hydroxystilbenes and derivatives thereof, such as resveratrol and 3,3′,5,5′-tetrahydroxystilbene (these anti-glycation agents are described in patent applications FR 2 802 425, FR 2 810 548, FR 2 796 278 and FR 2 802 420, respectively), dihydroxystilbenes and derivatives thereof, polypeptides of arginine and of lysine such as the product sold under the name Amadorine® by the company Solabia, carcinine hydrochloride (sold by Exsymol under the name Alistin®), an extract of Helianthus annuus, for instance Antiglyskin® from Silab, wine extracts such as the extract of powdered white wine on a maltodextrin support sold under the name Vin blanc déshydraté 2F by the company Givaudan, thioctic acid (or alpha-lipoic acid), a mixture of extract of bearberry and of marine glycogen, for instance Aglycal LS 8777® from Laboratoires Serobiologiques, and an extract of black tea, for instance Kombuchka® from Sederma, and mixtures thereof.
Preferred anti-glycation agents that will be mentioned include extracts of blueberry (Vaccinium myrtillus) and extracts of black tea.
Agents for Stimulating the Synthesis of Dermal and/or Epidermal Macromolecules and/or for Preventing their Degradation
Among the active agents for stimulating the dermal macromolecules or for preventing their degradation, mention may be made of those acting:

    
    
        either on collagen synthesis, such as extracts of Centella asiatica, asiaticosides and derivatives thereof; synthetic peptides such as iamin, biopeptide CL or palmitoyl oligopeptide sold by the company Sederma; peptides extracted from plants, such as the soybean hydrolysate sold by the company Coletica under the trade name Phytokine®; rice peptides such as Nutripeptide® from Silab, methylsilanol mannuronate such as Algisium C® sold by Exsymol; plant hormones such as auxins and lignans; folic acid; and an extract of Medicago sativa (alfalfa) such as the product sold by Silab under the name Vitanol®; a peptide extract of hazelnut such as the product sold by to the company Solabia under the name Nuteline C®; and arginine;
        or on the inhibition of collagen degradation, in particular agents acting on the inhibition of metalloproteases (MMP) more particularly such as MMP 1, 2, 3 and 9. Mention may be made of: retinoids and derivatives, extracts of Medicago sativa such as Vitanol® from Silab, an extract of Aphanizomenon flos-aquae (Cyanophyceae) sold under the name Lanablue® by Atrium Biotechnologies, oligopeptides and lipopeptides, lipoamino acids, the malt extract sold by the company Coletica under the trade name Collalift®; blueberry or rosemary extracts; lycopene; isoflavones, derivatives thereof or plant extracts containing them, in particular extracts of soybean (sold, for example, by the company Ichimaru Pharcos under the trade name Flavosterone SB®), of red clover, of flax or of kakkon; an extract of lychee; Dipalmitoyl Hydroxyproline sold by SEPPIC under the name Sepilift DPHP®: Baccharis genistelloides or Baccharine sold by Silab, an extract of moringa such as Arganyl LS 9781® from Cognis; the sage extract described in patent application FR-A-2 812 544 from the Labiatae family (Salvia officinalis from the company Flacksmann), an extract of rhododendron, a blueberry extract, and an extract of Vaccinium myrtillus such as those described in patent application FR-A-2 814 950;
        or on the synthesis of molecules belonging to the elastin family (elastin and fibrillin), such as: retinol and derivatives, in particular retinyl palmitate; the extract of Saccharomyces cerevisiae sold by the company LSN under the trade name Cytovitin®; and the extract of the alga Macrocystis pyrifera sold by the company Secma under the trade name Kelpadelie®; a peptide extract of hazelnut such as the product sold by the company Solabia under the trade name Nuteline C®;
        or on inhibition of elastin degradation, such as the peptide extract of seeds of Pisum sativum sold by the company LSN under the trade name Parelastyl®; heparinoids; and the N-acylamino amide compounds described in patent application WO 01/94381, such as {2-[acetyl(3-trifluoromethylphenyl)amino]-3-methylbutyrylamino}acetic acid, also known as N—[N-acetyl,N′-(3-trifluoromethyl)phenylvalyl]glycine, or N-acetyl-N-[3-(trifluoromethyl)phenyl]valylglycine or acetyl trifluoromethylphenylvalylglycine, or an ester thereof with a C1-C6 alcohol; an extract of rice peptides such as Colhibin® from Pentapharm, or an extract of Phyllanthus emblica such as Emblica® from Rona;
        or on the synthesis of glycosaminoglycans, such as the product of fermentation of milk with Lactobacillus vulgaris, sold by the company Brooks under the trade name Biomin Yoghurt®; the extract of the brown alga Padina pavonica sold by the company Alban Müller under the trade name HSP3®; the Saccharomyces cerevisiae extract available especially from the company Silab under the trade name Firmalift® or from the company LSN under the trade name Cytovitin®; an extract of Laminaria ochroleuca such as Laminaine® from Secma; essence of Mamaku from Lucas Meyer, and an extract of Cress (Odraline® from Silab);
        or on the synthesis of fibronectin, such as the extract of the zooplankton Salina sold by the company Seporga under the trade name GP4G®; the yeast extract available especially from the company Alban Müller under the trade name Drieline®; and the palmitoyl pentapeptide sold by the company Sederma under the trade name Matrixyl®.
    
    


Among the active agents for stimulating epidermal macromolecules, such as fillagrin and keratins, mention may be made especially of the extract of lupin sold by the company Silab under the trade name Structurine®; the extract of Fagus sylvatica beech buds sold by the company Gattefosse under the trade name Gatuline® RC; and the extract of the zooplankton Salina sold by the company Seporga under the trade name GP4G®; the copper tripeptide from Procyte; a peptide extract of Voandzeia substerranea such as the product sold by the company Laboratoires Sérobiologiques under the trade name Filladyn LS 9397®.
Preferably, an active agent that stimulates the synthesis of dermal and/or epidermal macromolecules and/or that prevents their degradation, chosen from agents for stimulating the synthesis of glycosaminoglycans, agents for inhibiting elastin degradation, agents for stimulating fibronectin synthesis, agents for stimulating the synthesis of epidermal macromolecules, and mixtures thereof, will be used.
Even more preferentially, an active agent that stimulates the synthesis of the glycosaminoglycans, chosen from an extract of the brown alga Padina pavonica, an extract of Saccharomyces cerevisiae, an extract of Laminaria ochroleuca, essence of Mamaku, and an extract of cress, and mixtures thereof, will be used.
As preferred active agents for stimulating the synthesis of dermal and/or epidermal macromolecules and/or for preventing their degradation, mention may be made of:
synthetic peptides such as iamin, the biopeptide CL or palmitoyloligopeptide sold by the company Sederma; peptides extracted from plants, such as the soybean hydrolysate sold by the company Coletica under the trade name Phytokine®; rice peptides such as Nutripeptide® from Silab, methylsilanol mannuronate such as Algisium C® sold by Exsymol; folic acid; an extract of Medicago sativa (alfalfa), such as the product sold by Silab under the name Vitanol®; a peptide extract of hazelnut, such as the product sold by the company Solabia under the name Nuteline C®; arginine; an extract of Aphanizomenon flos-aquae (Cyanophyceae) sold under the name Lanablue® by Atrium Biotechnologies, the malt extract sold by the company Coletica under the trade name Collalift®, lycopene; an extract of lychee; an extract of moringa such as Arganyl LS 9781® from Cognis; an extract of Vaccinium myrtillus such as those described in patent application FR-A-2 814 950; retinol and derivatives thereof, in particular retinyl palmitate; the extract of Saccharomyces cerevisiae sold by the company LSN under the trade name Cytovitin®; a peptide extract of hazelnut such as the product sold by the company Solabia under the name Nuteline C®; {2-[acetyl(3-trifluoromethylphenyl)amino]-3-methylbutyrylamino}acetic acid, also known as N—[N-acetyl,N′-(3-trifluoromethyl)phenylvalyl]glycine, or N-acetyl-N-[3-(trifluoromethyl)phenyl]valylglycine or acetyl trifluoromethylphenylvalylglycine, or an ester thereof with a C1-C6 alcohol; an extract of rice peptides such as Colhibin® from Pentapharm, or an extract of Phyllanthus emblica such as Emblica® from Rona; the extract of the brown alga Padina pavonica sold by the company Alban Müller under the trade name HSP3®; the extract of Saccharomyces cerevisiae available especially from the company Silab under the trade name Firmalift® or from the company LSN under the trade name Cytovitin®; an extract of Laminaria ochroleuca such as Laminaine® from Secma; the essence of Mamaku from Lucas Meyer, the extract of lupin sold by the company Silab under the trade name Structurine®; the extract of Fagus sylvatica beech buds sold by the company Gattefosse under the trade name Gatuline® RC.
Agents for Stimulating Fibroblast or Keratinocyte Proliferation and/or Keratinocyte Differentiation
The agents for stimulating fibroblast proliferation that may be used in the composition according to the invention may be chosen, for example, from plant proteins or polypeptides, extracted especially from soybean (for example a soybean extract sold by the company LSN under the name Eleseryl SH-VEG 8® or sold by the company Silab under the trade name Raffermine®); an extract of hydrolysed soybean proteins such as Ridulisse® from Silab; and plant hormones such as gibberellins and cytokinins; a peptide extract of hazelnut such as the product sold by the company Solabia under the name Nuteline C®.
Preferably, an agent that promotes keratinocyte proliferation and/or differentiation will be used.
The agents for stimulating keratinocyte proliferation that may be used in the composition according to the invention especially comprise: phloroglucinol, the extract of Hydrangea macrophylla leaves, for instance Amacha Liquid E® from Ichimaru Pharcos, a yeast extract such as Stimoderm® from CLR; the extract of Larrea divaricata such as Capislow® from Sederma, mixtures of extract of papaya, of olive leaves and of lemon, such as Xyleine® from Vincience, the extract of hydrangea macrophylla leaf such as Amacha liquid E® from Ichimaru Pharcos, retinol and esters thereof, including retinyl palmitate, the nut cake extracts sold by the Gattefosse and the extracts of Solanum tuberosum such as Dermolectine® sold by Sederma.
Among the agents for stimulating keratinocyte differentiation are, for example, minerals such as calcium; a peptide extract of lupin, such as the product sold by the company Silab under the trade name Structurine®; sodium beta-sitosteryl sulfate, such as the product sold by the company Seporga under the trade name Phytocohesine®; and a water-soluble extract is of corn, such as the product sold by the company Solabia under the trade name Phytovityl®; a peptide extract of Voandzeia substerranea such as the product sold by the company Laboratoires Sérobiologiques under the trade name Filladyn LS 9397®; and lignans such as secoisolariciresinol, and retinol and esters thereof, including retinyl palmitate.
As agents for stimulating keratinocyte proliferation and/or differentiation, mention may also be made of oestrogens such as oestradiol and homologues or cytokines.
As preferred active agents for stimulating fibroblast or keratinocyte proliferation and/or keratinocyte differentiation, mention will be made of plant proteins or polypeptides, extracted especially from soybean (for example a soybean extract sold by the company LSN under the name Eleseryl SH-VEG 8® or sold by the company Silab under the trade name Raffermine®); an extract of hydrolysed soybean proteins such as Ridulisse® from Silab; a peptide extract of hazelnut such as the product sold by the company Solabia under the name Nuteline C®; phloroglucinol, a yeast extract such as Stimoderm® from CLR; a peptide extract of lupin such as the product sold by the company Silab under the trade name Structurine®; a water-soluble corn extract, such as the product sold by the company Solabia under the trade name Phytovityl®; a peptide extract of Voandzeia substerranea, such as the product sold by the company Laboratoires Sérobiologiques under the trade name Filladyn LS 9397®; retinol and esters thereof, including retinyl palmitate.
Agents for Promoting the Maturation of the Horny Envelope
Agents that participate in the maturation of the horny envelope, which becomes impaired with age and induces a decrease in transglutaminase activity, may be used in the compositions of the invention. Examples that may be mentioned include urea and derivatives thereof and in particular Hydrovance® from National Starch and the other active agents mentioned in L'Oréal patent application FR 2 877 220.
NO-synthase Inhibitors
The agent with an inhibitory action on NO synthase may be chosen from OPCs (procyannidol oligomers); plant extracts of the species Vitis vinifera sold especially by the company Euromed under the name “Leucocyanidines de raisins extra”, or by the company Indena under the name Leucoselect®, or finally by the company Hansen under the name “Extrait de marc de raisin”; plant extracts of the species Olea europaea preferably obtained from olive tree leaves and sold especially by the company Vinyals in the form of a dry extract, or by the company Biologia & Technologia under the trade name Eurol® BT; and plant extracts of the species Gingko biloba, preferably a dry aqueous extract of this plant sold by the company Beaufour under the trade name “Ginkgo biloba extrait standard”, and mixtures thereof.
Agents for Stimulating the Energy Metabolism of Cells
The active agent for stimulating the energy metabolism of cells may be chosen, for example, from biotin, an extract of Saccharomyces cerevisiae such as Phosphovital® from Sederma, the mixture of sodium, manganese, zinc and magnesium salts of pyrrolidonecarboxylic acid, for instance Physiogenyl® from Solabia, a mixture of zinc, copper and magnesium gluconate, such as Sepitonic M3® from SEPPIC, and mixtures thereof; a beta-glucan derived from Saccharomyces cerevisiae, such as the product sold by the company Mibelle AG Biochemistry.
The invention also relates to a cosmetic skin treatment process for reducing or preventing the signs of ageing of the skin or its integuments (hair, eyelashes, nails, etc.), comprising at least one step that consists in applying to the skin at least one composition as defined previously.
The process according to the invention more specifically comprises at least one step that consists in applying, to the skin of individuals whose skin shows at least one of the signs of cutaneous ageing recalled previously, at least one composition as defined previously.
More particularly, it comprises at least one step that consists in applying a composition as defined previously to the skin of individuals having skin or an area of skin that is aged, wrinkled, flabby and/or flaccid, or to areas of the body showing a lack of elasticity and/or firmness and/or tonicity.
The composition according to the invention may be applied to the part of the skin or integuments to be treated, in particular to the face, the body, the neck, the hands, the hair or the scalp, preferably daily or several times a day. The application may, for example, be repeated every day over a variable period according to the desired effects, generally from 3 to 6 weeks, but may be prolonged or pursued continuously.
According to one alternative, the composition according to the invention may be administered by injection optionally in combination with filling products. Specifically, one of the solutions adopted for combating wrinkles and/or the loss of volume of soft tissue is the use of filling products (or filler). This filling may be achieved by using non-resorbable products, such as polyacrylamide gels or polymethyl methacrylate (PMMA) particles. However, these compounds may lead to intolerance reactions of the type such as inflammation or hypersensitivity.
The use of resorbable components, such as proteins, fats, collagen or hyaluronic acid, is preferred. However, these compounds are degraded relatively quickly in the body, which reduces their efficacy. To overcome this, more or less expensive crosslinking of these components must be performed. At the present time, the hyaluronic acid used in pharmaceutical forms or medical devices is in the form of a sodium hyaluronate gel. The monosaccharide according to the invention or the compositions containing it may also be applied by mesotherapy. Mesotherapy is a technique of treatment via intraepidermal and/or intradermal and/or subcutaneous injection of active product(s), for instance micronutrients, vitamins and/or hyaluronic acid. The compositions are administered according to this technique via injection in the form of multiple small droplets into the epidermis, the dermo-epidermal junction and/or the dermis in order especially to perform subcutaneous layering. The mesotherapy technique is especially described in the publication “Traité de mésothérapie” by Jacques Le Coz, published by Masson, 2004. Mesotherapy performed on the face is also referred to as a mesolift or a mesoglow.
Thus, another subject of the present invention may be a device, in particular a medical device, comprising an effective amount of at least one monosaccharide as defined previously, in combination with an effective amount of at least one ascorbic acid analogue, or ascorbic acid itself. This device may be suitable for intraepidermal and/or intradermal and/or subcutaneous injection. The combination of active agents as defined above is dissolved in a sterile medium. The said device may comprise at least one other compound, for instance at least one resorbable or non-resorbable product, such as those mentioned above, which is optionally crosslinked.
The said device may be, for example, a syringe with a needle or an injection device without a needle, such as those used in the care technique known as mesotherapy. A kit comprising a device may also be envisaged, the said kit comprising a device, in particular a syringe or an injection device, and at least a combination of active agents, monosaccharide(s) and ascorbic acid or analogue(s), as defined above. The said kit may also comprise a needle. The said device may be in ready-to-use form, i.e. prefilled, or may need to be filled before use. In the latter case, a composition or another device (such as a vial) comprises the said combination of active agents, monosaccharide(s) and ascorbic acid or analogue(s), optionally in combination with at least one other active compound, for instance at least one resorbable or non-resorbable product, such as the filling products mentioned above, which is optionally crosslinked.
The injection of the combination according to the invention may be performed simultaneously with, or before or after, the application to the skin or the integuments of another cosmetic or pharmaceutical composition, preferably a dermatological composition, comprising, in a physiologically acceptable support, at least one other active agents, as mentioned above.
According to another aspect, the invention also relates to a cosmetic assembly comprising: i) a container delimiting at least one compartment, the said container being closed by a closing member; and ii) a composition as defined previously, placed inside the said compartment.
The container may be in any suitable form. It may especially be in the form of a bottle, a tube, a jar, a case, a can, a sachet or a box. The closing member may be in the form of a removable stopper, a lid, a cover, a tear-off strip or a cap, especially of the type comprising a body fixed to the container and a cap articulated on the body. It may also be in the form of a member ensuring the selective closure of the container, especially a pump, a valve or a clapper.
The container may be combined with an applicator. The applicator may be in the form of a fine brush, as described, for example, in patent FR 2 722 380. The product may be contained directly in the container, or indirectly. By way of example, the product may be arranged on an impregnated support, especially in the form of a wipe or a pad, and arranged (individually or in plurality) in a box or in a sachet. Such a support incorporating the product is described, for example, in patent application WO 01/03538.
The closing member may be coupled to the container by screwing.
Alternatively, the coupling between the closing member and the container is done other than by screwing, especially via a bayonet mechanism, by click-fastening, gripping, welding, bonding or by magnetic to attraction. The term “click-fastening” in particular means any system involving the crossing of a bead or cord of material by elastic deformation of a portion, especially of the closing member, followed by return to the elastically unconstrained position of the said portion after the crossing of the bead or cord.
The container may be at least partially made of thermoplastic material such as polypropylene or polyethylene.
Alternatively, the container is made of non-thermoplastic material, especially glass or metal (or alloy).
The container may have rigid or deformable walls, especially in the form of a tube or a tube bottle. The container may comprise means for initiating or facilitating the distribution of the composition. By way of example, the container may have deformable walls so as to allow the composition to exit in response to a positive pressure inside the container, this positive pressure being caused by elastic (or non-elastic) squeezing of the walls of the container.
The contents of the patents or patent applications mentioned previously are incorporated by reference into the present patent application.
According to one particular mode, the invention relates to a cosmetic assembly comprising:

    
    
        a composition A containing at least one compound chosen from ascorbic acid, an analogue thereof and mixtures thereof,
        a composition B, conditioned separately from composition A, comprising at least one monosaccharide chosen from mannose, rhamnose and a mixture thereof.
    
    


Finally, the invention relates to a cosmetic or dermatological treatment process comprising at least one step of administration, in particular of topical application, to the skin and/or its integuments, of composition A and at least one step of administration, in particular of topical application to the skin and/or its integuments, of composition B.
The administration of composition A according to the invention may be performed simultaneously with, or before or after, the administration of composition B. As specified previously, the administration of composition A and of composition B may be performed topically, orally or via injection.
According to one alternative, composition A is administered first and composition B is administered second. According to another alternative, composition B is administered first and composition A is administered second.
Compositions A and B may be conditioned separately inside two compartments, formed either by two separate containers, or inside a single device. The term “single device” means a device via which the two compartments are solidly attached. Such a device may be obtained via a process of monobloc moulding of the two compartments, especially made of a thermoplastic material. It may also result from any form of assembly, especially by bonding, welding or other click-fastening.
According to a first embodiment, the two containers are independent of each other. Such containers may be in various forms. They may especially be tubes, bottles or drums.
One and/or the other of the containers may be fitted with a manually operated pump on which is mounted a push button for actuating the pump and dispensing the composition via at least one dispensing orifice.
Alternatively, one and/or the other of the containers is pressurized, especially by means of a propellant, in particular a propellant gas. In this case, the container(s) is (are) equipped with a valve on which is mounted a push-button equipped with a nozzle or any other diffusion means for dispensing the product.
The propellant may be in a mixture with the composition to be dispensed or separated, especially via a piston that can slide inside the container, or via the flexible walls of a bag inside which the composition is placed.
The containers may be made of various materials: plastic, glass or metal.
Alternatively also, the two compartments are formed from two concentric compartments formed inside a tube, and mounted thereon is a pump with no air reuptake, and equipped with a push button with one or two dispensing orifices. Provided inside the tube is a piston that rises in the direction of the pump as and when the compositions are withdrawn from inside the containers. Such dispensing modes are especially used for dispensing toothpastes.
Key to the Figures
FIG. 1: Diagram schematically representing the results obtained for the keratinocyte proliferation, in the presence of a control, in the presence of different markers, in medium deficient in growth factors, and with addition of different concentrations of L-rhamnose reported on the x-axis. The values reported on the y-axis correspond to the percentages of labelled cells measured relative to the control.
FIG. 2: Diagram schematically representing the results obtained for the keratinocyte proliferation, in the presence of a control, in the presence of different markers, in medium deficient in growth factors, and with addition of different concentrations of D-mannose reported on the x-axis. The values reported on the y-axis correspond to the percentages of labelled cells measured relative to the control.
FIG. 3: Diagram representing the number of fibroblasts measured between an untreated control whole reconstructed skin, on the left, and a whole reconstructed skin treated with 5 mM of rhamnose, on the right. The fibroblasts are counted at different stages of the treatment. Thus, for each skin type, the left-hand column corresponds to the count obtained at 48 hours and the right-hand column corresponds to the count obtained at 120 hours of treatment.
FIG. 4: Photographs of frozen sections of reconstructed skin 7 μm thick. The level of fluorescence is materialized by the white marks on the black and white photograph; it is proportional to the amount of type I procollagen. The control skin is on the left, and skin treated with 1 mM of rhamnose is on the right.
FIG. 5: Diagram representing the amount of collagens VI and XII synthesized by dermal fibroblasts in the presence or absence of rhamnose and/or vitamin C.
The invention is illustrated in greater detail in the examples that follow, which are given as non-limiting illustrations of the field of the invention.
EXAMPLES
Example 1
Proliferation of Keratinocytes
Protocol
The keratinocytes (HaCat line) are cultured under two conditions: whole defined culture medium (standard condition) and culture medium deficient in growth factors. This deficient medium gives rise to a controlled delay in cell proliferation. Under these conditions, it is then possible to measure the effects of compounds capable of compensating for the deficiency in growth factors of the culture medium and thus of relaunching the cell multiplication and/or of stimulating cell metabolism.
The keratinocyte proliferation is measured by means of three markers on the same cell population: the level of DNA, which is proportional to the number of cells (Cyquant probe), the level of constituent polar lipids of cell membranes (Nile red probe) and the mitochondrial respiration, which reflects the general cell metabolism (XTT probe).
Results
The results are given in FIGS. 1 and 2.
The two monosaccharides rhamnose and mannose demonstrate their capacity to activate keratinocyte proliferation when the keratinocytes are cultured in medium depleted in growth factors, a culturing condition that significantly delays their cell growth.
This activation of cell proliferation by the two compounds is manifested by a higher number of cells when compared with the untreated control.
This increased number of cells is materialized by a level of DNA (Cyquant), a level of polar lipids (Nile red signal) and a mitochondrial respiration (XTT signal) that are significantly increased when the monosaccharides are evaluated at 1 mM. At 500 μM, the two molecules already show efficacy. The two monosaccharides mannose and rhamnose thus exert an influence on keratinocyte proliferation. They activate the proliferation of keratinocytes cultured in medium depleted in growth factor, which is manifested by a higher number of cells when compared with an untreated control.
Rhamnose and mannose thus show anti-ageing efficacy by boosting epidermal renewal and combating age-related epidermal atrophy.
Example 2
Proliferation of Fibroblasts
Protocol
Rhamnose was studied on a model of whole reconstructed skin in order to measure its anti-ageing efficacy on the dermal compartment.
Briefly, the model of reconstructed skin used is that described by Bell et al. (Bell E. et al., The reconstitution of living skin, J. Invest. Dermatol., 1983, July; 81): it includes a dermal equivalent on which is reconstructed a multistratified epidermis; the dermal equivalent is manufactured from acid-soluble collagen, culture medium containing serum and normal adult human fibroblasts. After 5 days of shrinkage, this equivalent is inoculated with keratinocytes and then cultured for 6 days in immersion and for 7 days in emersion in order to obtain a multistratified and differentiated epidermis having a horny layer.
The reconstructed skin is treated with 5 mM rhamnose for 2 days and 5 days in the culture medium; after the treatment, the reconstructed skins are included in Tissue Tek in order to produce frozen sections 7 μm thick with a cryostat. The sections produced are then stained with propidium iodide to label the DNA of the nuclei of the fibroblasts in order to count them. Three frozen sections are prepared at random on each reconstructed skin; on each section, two microscopic fields (25× objective lens) are analysed by fluorescence microscopy and photographed. The dermal fibroblasts are thus counted for each reconstructed skin on six images in total representing the six microscopic fields considered. The number of dermal fibroblasts is compared between the control skin and that treated with rhamnose at the two kinetic stages.
Results
The results are given in FIG. 3.
It was found that rhamnose induces stimulation of growth of the dermal fibroblasts of the reconstructed skin within 48 hours of treatment, this stimulation being confirmed at 120 hours of treatment, with between 30% and 35% additional cells (see FIG. 3). It should be noted that this stimulation is accompanied by a stimulation of procollagen 1 synthesis at 5 mM, and also at 1 mM, which may also result from the increased number of fibroblasts responsible for the secretion of this major protein of the extracellular matrix.
These two effects complement the anti-ageing activity of rhamnose already measured on the epidermal compartment, by stimulating the proliferation and metabolism of the fibroblast, which is a major cell of the dermal compartment.
Example 3
Synthesis of Procollagen 1
Conventional detection via indirect immunofluorescence of type I procollagen in the dermis of the reconstructed skin was also performed on other series of frozen sections (anti-procoll 1 antibody (MAB 1912 Millipore)+FTIC-coupled conjugate (112-095-068 Jackson Immunoresearch)). In order to obtain bearings within the cutaneous architecture during the microscopic examination of the sections, the cell nuclei of the keratinocytes and fibroblasts are localized by staining them with propidium iodide, as described above. Three frozen sections are prepared at random on each reconstructed skin and on each section, and two microscopic fields (25× objective lens) are analysed by fluorescence microscopy and photographed. The levels of fluorescence proportional to the amount of type I procollagen are compared between the control skin and the skin treated with rhamnose.
In image 1, FIG. 4, corresponding to a section of control reconstructed skin at 120 hours of culture, the presence of type 1 procollagen synthesized by the dermal fibroblasts is materialized by the green fluorescence located in the bottom part of the image. The basal part of the epidermis, highly cellular tissue, which may be visualized by the numerous keratinocyte nuclei, can be made out in the top part of the image. The dermis, much less cellular tissue, also reveals the random distribution of the fibroblasts within the dermal extracellular matrix. In image 2, FIG. 4, corresponding, for example, to a section of reconstructed skin treated with 1 mM rhamnose for 120 hours, a marked increase in green fluorescence is noted when compared with that observed for the control skin (image 1), and also a distribution of the fluorescent signal clearly materializing the fibrillar aspect of the newly synthesized type I procollagen. This increase in general fluorescence indicates that the rhamnose treatment has greatly stimulated the synthesis of type I procollagen by the fibroblasts.
These results clearly show the capacity of rhamnose to stimulate fibroblast metabolism, which metabolism, in the course of ageing, becomes more imbalanced towards degradation of the extracellular matrix than towards its renewal.
By stimulating both the metabolism and growth of dermal fibroblasts, rhamnose clearly demonstrates its anti-ageing efficacy on the dermis, this efficacy being complementary to that measured with respect to the epidermal compartment.
Example 4
Combination of Rhamnose and Ascorbic Acid, Demonstration of the Synergism of Action of Vitamin C and Rhamnose on the Synthesis of Epidermal Ceramides
Protocol
For Ascorbic Acid Alone (without Rhamnose):
Ascorbic acid is tested on a skin equivalent sold by the company Episkin (Lyons, France) after culturing it for 7 days. The procedure and the amounts of ascorbic acid used are identical to the protocol detailed hereinbelow for the combination with rhamnose.
For the Combination of Active Agents According to the Invention:
The combination of ascorbic acid and rhamnose is tested on a skin equivalent sold by the company Episkin (Lyons, France), after culturing it for 7 days. The culture media and test media are those included in the kit sold by the supplier. The ascorbic acid was tested at 100 and/or 200 μg/ml in combination with rhamnose at 5 mM in the culture medium. The samples of reconstructed epidermis are treated for 24 hours. The control is constituted by an identical epidermal equivalent not subjected to any treatment. The reconstructed epidermis is incubated overnight with 14C acetate (2 μCi/ml), to monitor the lipid synthesis. At the end of the incubation, the epidermal equivalent is detached from its collagen support.
The preparation of the lipids from the epidermal equivalent and their analysis by HPTLC (high-performance thin layer chromatography) are performed according to the technique and with the buffers described by M. Ponec (1991, Adv. Lipid Res. 24: 83-117).
At the end of migration, densitometric analysis of the autoradiography is performed using a model 1800, Fuji brand densitometer. The amounts of total epidermal lipids between the samples of control epidermis and those treated with the combination are compared. As a supplement, the modulation of the specific classes of lipids, for instance the phospholipids, free fatty acids, ceramides and cerebrosides, and cholesterol, may also be analysed more particularly.
Results
A production of epidermal lipids of superior amount is noted when the samples of reconstructed epidermis are treated with the combination of ascorbic acid and rhamnose, compared with ascorbic acid alone.
Example 5
Combination of Rhamnose and Ascorbic Acid, Demonstration of the Synergism of Action of Vitamin C and Rhamnose on the Synthesis of Collagens VI and XII by Fibroblasts
This example describes the synergic effect of the association of vitamin C with L-rhamnose on the expression of the alpha 1-chain of collagens VI and XII of the extracellular matrix.
Protocol
Human dermal fibroblasts (used at the ninth passage) are obtained from mammary or abdominal surgeries. The culture medium implemented is DMEM, supplemented with:


    
    
        fetal calf serum (FCS), 10%
        L-Glutamine, 2 mM (Invitrogen 25030024),
        Penicillin (50 UI/mL), streptomycin (50 μg/mL), (Invitrogen 15070063),
        Sodium pyruvate (Invitrogen 11360039), and
        Non essential amino acids (Invitrogen 11140035).

Fibroblasts are cultured (D0) in supplemented DMEM medium as described above, incubated at 37° C. znd 5% CO2 in an atmosphere saturated with water. After adhesion of the cells (D1), the culture medium is removed and replaced with medium containing vitamin C, L-rhamnose or their association. The different stimulation conditions (vitamin C 25 μM, 50 μM, Rhamnose 1 mM and the different associations) were performed in triplicate (12 well dish). Studies of the expression of the different collagenes were performed on D3 and D5.

Extraction and Purification of RNAs:

For each condition and at each time (D3 and D5), the supernatants are removed and the cell lawns are lysed. Total RNAs are then extracted and purified. All these operations are performed by implementing the Qiagen to n° 79254 kit.

Quantification and Qualification of RNAs:

RNAs are then quantified in fluorescence according to the Ribogreen protocol (Quan-it, Ribogreen RNA assay kit, invitrogen R11490) and reading with Tecan Safire. The quality of RNAs is appreciated by following the electrophoretic profile.

Final tested concentrations of vitamin C, rhamnose and their associations are the following: Vitamin C 50 μM, L-Rhamnose 1 mM, and association: Vitamin C 50 μM+L-Rhamnose 1 mM.

    
    


Results














Collagen VI
Collagen XII



Col6A1
Cola12A1


















Control
100
100


Rhamnose 1 mM
109
115


Vitamin C 50 μM
154
124


Rhamnose 1 mM + Vitamin C 50 μM
174
152









The table above presents the intensity of the biosynthesis of collagenes VI and XII in fibroblasts in presence of rhamnose, vitamin C or their association compared to the control that is at 100.
The results are also represented in FIG. 5. They demonstrate a synergism of action of rhamnose and vitamin C in regulating the transcription of genes coding for collagens VI and XII by dermal fibroblasts.
Example 6
Example of Preparation of a Cosmetic Composition According to the Invention










Epidermal and dermal regenerating creams: oil-in-water emulsion
















Ammonium Polyacryldimethyltauramide (Hostacerin AMPS
1.00%


from Clariant)


Cyclohexasiloxane
 5.0%


Apricot kernel oil
  7%


Isononyl isononanoate
  7%


Stearyl alcohol
0.30%


Glyceryl stearate/PEG-100 stearate
0.70%


Dimyristyl tartrate/cetearyl alcohol/C12-15 pareth-7/PPG-
0.50%


25 laureth-25


Xanthan gum
0.20%


Rhamnose
  5%


Ascorbic acid
  3%


Preserving agents
0.50%


Water
qs 100









Example 7
Example of Preparation of a Cosmetic Composition According to the Invention










Epidermal regeneration creams: oil-in-water emulsion


















Ammonium Polyacryldimethyltauramide (Hostacerin
1.00%



AMPS from Clariant)



Cyclohexasiloxane
 5.0%



Glycerol
1.70%



Stearyl alcohol
0.30%



Apricot kernel oil
  7%



Isononyl isononanoate
  7%



Dimyristyl tartrate/cetearyl alcohol/C12-15
0.50%



pareth-7/PPG-25 laureth-25



Xanthan gum
0.20%



Ascorbic acid
  5%



SMA 1000 HNa
  1%



Mannose
  5%



Reserving agents
0.50%



Water
qs 100










Example 8
Example of Preparation of a Cosmetic Composition According to the Invention










Epidermal regeneration creams: oil-in-water emulsion


















Ammonium Polyacryldimethyltauramide (Hostacerin
1.00%



AMPS from Clariant)



Cyclohexasiloxane
 5.0%



Glycerol
1.70%



Stearyl alcohol
0.30%



Apricot kernel oil
  7%



Isononyl isononanoate
  7%



Dimyristyl tartrate/cetearyl alcohol/C12-15
0.50%



pareth-7/PPG-25 laureth-25



Xanthan gum
0.20%



Mannose
 2.5%



Rhamnose
 2.5%



Magnesium ascorbyl phosphate
  1%



Preserving agents
0.50%



Water
qs 100










Example 9
Example of Preparation of a Cosmetic Composition According to the Invention
Anti-ageing Facial Day Cream












Phase A1:











Sucrose distearate sold by the company
1.75%




Stéarinerie Dubois



Sorbitan stearate oxyethylenated with 4 mol



of ethylene oxide, sold by the company



ICl under the name Tween 61
1.15%



Stearic acid
0.75%



Stearyl heptanoate
4.00%



Petroleum jelly codex
1.50%



Avocado oil
3.20%



Jojoba oil
3.00%



Volatile silicone oil
2.70%



Vitamin E acetate
1.00%



Vitamin F glycerides
3.00%







Phase A2:











Silicone gum sold by Dow Corning under
3.00%




the name Q2-1403 Fluid



Propyl paraben
0.2%



Fragrance
0.3%







Phase B:











Glycerol
3.00%




Hydroxyproline
1.00%



D-Panthenol
1.00%



Triethanolamine
0.35%



Rhamnose
3.00%



Glycosyl vitamin C
1.00%



Methyl paraben
0.3%



Demineralized water
qs 100%







Phase C:











Ammonium Polyacryldimethyltauramide
1%




(Hostacerin AMPS from Clariant)










The above written description of the invention provides a manner and process of making and using it such that any person skilled in this art is enabled to make and use the same, this enablement being provided in particular for the subject matter of the appended claims, which make up a part of the original description.
As used herein, the phrases “selected from the group consisting of,” “chosen from,” and the like include mixtures of the specified materials. Terms such as “contain(s)” and the like as used herein are open terms meaning ‘including at least’ unless otherwise specifically noted. The term “mentioned” notes exemplary embodiments, and is not limiting to certain species. As used herein the words “a” and “an” and the like carry the meaning of “one or more.”
All references, patents, applications, tests, standards, documents, publications, brochures, texts, articles, etc. mentioned herein are incorporated herein by reference. Where a numerical limit or range is stated, the endpoints are included. Also, all values and subranges within a numerical limit or range are specifically included as if explicitly written out.
The above description is presented to enable a person skilled in the art to make and use the invention, and is provided in the context of a particular application and its requirements. Various modifications to the preferred embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments and applications without departing from the spirit and scope of the invention. Thus, this invention is not intended to be limited to the embodiments shown, but is to be accorded the widest scope consistent with the principles and features disclosed herein. In this regard, certain embodiments within the invention may not show every benefit of the invention, considered broadly.

## Claims

1. A method, comprising applying to the skin or its integuments a composition comprising, in a physiologically acceptable medium, a combination of the following two active agents: 1) at least one monosaccharide, wherein the monosaccharide is rhamnose, and wherein the at least one monosaccharide is present as a monomer, and 2) at least one additional compound selected from the group consisting of ascorbic acid, an ascorbic acid salt, the acetic ester of ascorbic acid, a saccharide ester of ascorbic acid, and a metal salt of phosphoryl ascorbic acid, and in which the amount of the monosaccharide(s) is 0.4%-30% by weight relative to the total weight of the composition, wherein said method is a method for at least one selected from the group consisting of:
reducing the signs of ageing of skin, and
reducing wrinkles and/or fine lines, and
wherein said composition is applied to human skin in need thereof in an amount effective to increase an epidermal lipid profile by modifying lipogenesis and to increase synthesis of collagen VI and/or collagen XII.

2. The method according to claim 1, wherein said method is a method for treating wrinkles and/or fine lines, wherein said composition is applied to human skin in need thereof.

3. The method of claim 1, wherein the additional compound is at least one selected from the group consisting of an ascorbic acid salt, the acetic ester of ascorbic acid, a saccharide ester of ascorbic acid, and a metal salt of phosphoryl ascorbic acid.

4. The method of claim 3, wherein the additional compound is magnesium ascorbyl phosphate.

5. The method of claim 1, wherein the additional compound is at least one saccharide ester of ascorbic acid selected from the group consisting of glycosyl, mannosyl, fructosyl, fucosyl, galactosyl, N-acetylglucosamine and N-acetylmuramic esters of ascorbic acid.

6. The method of claim 1, wherein the additional compound is at least one saccharide ester of ascorbic acid selected from the group consisting of ascorbyl-2 glucoside, 2-O-α-D-glucopyranosyl of L-ascorbic acid and 6-O-D-galactopyranosyl of L-ascorbic acid.

7. The method of claim 1, wherein the additional compound is at least one metal salt of phosphoryl ascorbic acid selected from the group consisting of alkali metal ascorbyl phosphates, alkaline-earth metal ascorbyl phosphates, and transition metal ascorbyl phosphates.

8. The method of claim 1, wherein the amount of additional compound present in the composition is 0.001%-30% by weight relative to the total weight of the composition.

9. The method of claim 1, wherein the composition is suitable for topical administration to the skin or its integuments, oral administration, and/or cutaneous injection.

10. The method of claim 1, wherein the additional compound is ascorbic acid.

11. The method of claim 1, wherein the increase in epidermal lipid profile by modifying lipogenesis and increase in synthesis of collagen VI and/or collagen XII is a material increase as compared to the amount of epidermal lipid profile by modifying lipogenesis and synthesis of collagen VI and/or collagen XII in a composition containing the additional compound but lacking the monosaccharide.