FR3007982A1 - STABILIZATION OF DIFFERENTIATED CELLS OF ROSE. - Google Patents

Abstract

Use of a metal ion chelating agent as a stabilizing agent in a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof.

Description

FIELD OF THE INVENTION The present invention relates to the use of chelating agents for the stabilization of a composition comprising dedifferentiated cells of a plant of the species Rosa sp.

More particularly, the use of a metal ion chelating agent makes it possible to stabilize a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or a lyophilizate thereof ART PRIOR ART The use of dedifferentiated plant cells for cosmetic purposes is a concept that has emerged over the last twenty years or so. The dedifferentiated plant cells were born from Haberland's work in 1902. During the last 40 years, plant cell cultures have been used for the production of metabolites of interest or for the propagation of plants in the same way (somatic embryogenesis). This plant biotechnology is based on the concept of "cell totipotency", which proposes that any plant cell be able to dedifferentiate and regenerate another individual identical to the one from which it originates. A dedifferentiated plant cell can be obtained from a plant cell from a differentiated organ, such as, for example, leaf, stem, root, petal. When this differentiated cell is cultured, it can return to a dedifferentiated state, that is to say that it loses its cellular specificity of leaf, stem, root or petal, and eventually becomes capable of generating the whole plant. . A dedifferentiated plant cell is the equivalent of a true plant stem cell derived from meristematic cells of plants, and having no specific biological past of an organ. WO 2009/151302 and KR 2009-0118877 disclose anti-aging or antioxidant compositions containing undifferentiated plant cells, derived from the cambium of Panax ginseng or a plant of the genus Taxus.

EP 1 985 280 describes the use of dedifferentiated plant cells derived from a plant of the family Rosaceae, and in particular apple tree Malus domestica, to protect the stem cells of the skin against various intrinsic and extrinsic stresses, and in particular to treat skin disorders or age-related hair.

EP 1 699 423 describes the use of a dedifferentiated plant cell lyophilizate from a halophilic plant to rejuvenate the appearance of the skin. DE-A-102 009 027 361 and EP 2 266 529 describe the use of dedifferentiated plant cells, in particular derived from a plant of the genus Malus, for treating the keratin fibers of the hair, and in particular protecting them against UV radiation, or for the treatment of aged skin. Finally, EP 0 909 556 describes the use of an extract of a plant of the Rosaceae family, in particular a rose, optionally obtained from undifferentiated plant cells, as a bradykinin antagonist for the treatment of various diseases. , especially cutaneous. The inventors have observed that dedifferentiated rose plant cells, or an extract or lyophilizate thereof, have proved particularly suitable and effective for treating aged skin or hair. However, the inventors have observed that compositions comprising dedifferentiated rose plant cells, and in particular compositions comprising dedifferentiated rose cells obtained by a process comprising a freezing then thawing step or a lyophilization step, have a reduced stability at course of time. The Applicant has observed that with storage time, a composition comprising plant cells dedifferentiated with pink or comprising an extract or a lyophilizate thereof, has a yellowish coloration or brownish coloration, and may contain color aggregates black. These characteristics, which materialize an alteration of the composition over time, appear a few hours to a few days after obtaining the composition. In particular, these features of yellowing of the composition, of browning of the composition, or of the formation of black-colored aggregates, appear a few hours to a few days after the composition is obtained by a process comprising a step of freezing then thawing. These visible changes in the composition over time mean reduced stability of the latter, with a deterioration which increases with the storage time.

This degradation of the visual appearance, which would result from a change of color, a change in texture that would result or not the presence of aggregates, or a combination of several of these alterations, is incompatible with a formulation of the composition in a cosmetic product.

In particular, these changes in color and / or texture of a composition comprising dedifferentiated cells of a plant of the species Rosa sp. or an extract or a lyophilizate thereof, signify the occurrence of a physicochemical alteration of the composition, these changes being able to reduce the cosmetic effect (s) thereof with the increase of the storage duration . There is therefore a need to identify technical solutions for increasing the stability properties of compositions comprising dedifferentiated cells of a plant of the Rosa sp species, or comprising an extract or lyophilizate thereof. More particularly, there is a need to confer a long-term preservation of the texture homogeneity of a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof. ci, that is to say a need to delay, or to reduce, or to block the formation of aggregates of plant material. There is also a need to maintain over time the visual appearance of a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or a lyophilizate thereof, it is that is, a need to retard, or to reduce, or to block the yellowing or browning of the composition. SUMMARY OF THE INVENTION The present invention relates to the use of a metal ion chelating agent as a stabilizing agent in a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof. In another aspect, the invention relates to a method for stabilizing a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof, which method comprises at least a step of contacting said dedifferentiated cells, said extract or said lyophilizate thereof with a metal ion chelating agent. Another aspect of the invention relates to a stable composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof, said composition comprising, as an agent stabilizer, a metal ion chelating agent.

According to another aspect, the invention relates to the cosmetic use of a stable composition as defined in the present description, as an active agent for the care of aged skin or aged hair.

LEGEND OF THE FIGURE Figure 1: Photon microscopy photos of dedifferentiated plant cell compositions of rose after storage at room temperature. FIG. 1A: magnification (x10) microscopy plate in unpolarized light of a Lancôme Rose cell composition obtained according to a process comprising a step of freezing then thawing. FIG. 1B: magnification (x10) microscope slide in polarized light of a composition of Lancôme Rose cells obtained according to a process comprising a step of freezing then thawing.

DETAILED DESCRIPTION OF THE INVENTION Surprisingly, the inventors have shown that the addition of a metal ion chelating agent, such as, for example, EDTA or an EDTA salt, makes it possible to stabilize a composition comprising cells. dedifferentiated from a plant of the species Rosa sp., or comprising an extract or lyophilizate thereof. It is thus shown according to the invention that the addition of a metal ion chelating agent, such as, for example, EDTA or an EDTA salt, makes it possible to maintain a homogeneous texture, without visible deterioration of the appearance of origin of said composition during storage time. For the purposes of the invention, the term "stabilization" means that the composition, during its storage or storage, retains a visual appearance, especially its color, and a texture identical or similar to that of the freshly obtained composition by the method, that is to say a homogeneous texture and a colorless or slightly whitish visual appearance. The presence of a metal ion chelating agent in the composition has the effect of retarding, reducing, or blocking the formation of aggregates of plant material, which includes exposed visible aggregates and aggregates visible only by observation by light microscopy. Thus, with a composition comprising a metal ion chelating agent as defined in the present description, there is an absence of aggregates of plant material, or at least the presence of a reduced amount of aggregates is visible after observation by optical microscopy.

The presence of a metal ion chelating agent in the composition also has the effect of delaying, reducing, or blocking the yellowing or browning of the composition, as can be observed directly by naked or photon microscopy.

Due to the absence of aggregates of plant material, or because of the presence of a reduced amount of aggregates of plant material, and because of the absence of detectable yellowing or browning, the visual appearance and texture of the composition is identical or similar to the original appearance of the composition, that is to say to the visual appearance and texture of the composition such as immediately resulting from the process used to prepare it.

For the purposes of the invention, the term "dedifferentiated plant cell" means any cell strain derived from organs of a rose bush of the species Rosa sp. and obtained by specific in vitro culture conditions, no longer having any character of specialization and capable, under the effect of an induction, to any differentiation according to their genome and to generate by itself an entire plant of plant of which it comes from. Such cells are able to live on their own and not in dependence on other cells. Dedifferentiated plant cells are therefore distinct from undifferentiated plant cells that exist naturally in plants. Under normal conditions the plant cells express about 20% of their genome, the remaining 80% being expressed only in response to particular environmental conditions. In vitro culturing of these cells under particular culture conditions makes it possible to "reprogram" the cells and thus to access a part of this non-expressed genome. Some compounds, difficult to obtain by plant extractions, become more accessible in cell cultures. Thus, advantageously, the plant dedifferentiated cells allow access to new compounds not present in the whole plant or significantly increase the expression of known molecules but found in very small amounts in the whole plant. Dedifferentiated plant cells can be obtained from any plant of the species Rosa sp. These cells can be obtained from plant material derived from whole plants or parts of plants such as leaves, stems, flowers, petals, sepals, or roots grown in vivo or in vitro. By in vivo culture is meant any culture of conventional type, ie in soil in the open air or in the greenhouse or above ground.

By in vitro culture is meant all the techniques known to those skilled in the art which artificially allows the production of a plant or part of a plant. The selection pressure imposed by the physicochemical conditions during the growth of the plant cells in vitro makes it possible to obtain a standardized plant material that is available throughout the year, unlike plants grown in vivo. Preferably, according to the invention, a plant derived from an in vivo culture is used. Preferably, dedifferentiated plant cells of the invention are obtained from at least one leaf of a plant of the species Rosa sp. The Rosa genus contains more than 1000 species, among which Rosa alba, Rosa alpina, Rosa canina, Rosa cinnamonea, Rosa gallica, Rosa repens, Rosa rubrifolia, Rosa rubiginosa, Rosa sempervirens, Rosa spinosissima, Rosa stylosa, Rosa tomentosa, or Rosa villosa. Still more preferably, the dedifferentiated plant cells of the invention are obtained from a Rose Lancôme rosebush, and more particularly from a Rose Lancôme delboip rosebush. Such a rosebush is commercially available, for example, from Delbard. The Lancôme rosebush is a tea hybrid with a female hybrid, a cross between Dr. Albert Schweitzer x [Michèle Meilland x Bayadèrel and cross-pollinated by the variety Melmet. According to the culture method chosen, and in particular according to the selected culture medium, it is possible to obtain, from the same explant, dedifferentiated plant cells having different characters. A culture medium suitable for obtaining dedifferentiated plant cells of the invention is well known to those skilled in the art and can include NH4NO3; KNO3C12Cl2.2H2O; MgSO4; KH2PO4; MnSO4.4H2O; ZnSO4.7H2O; KI; Na2MoO4.2H2O; CuSO4.5H2O; FeSO4,7H2O; myo-inositol; nicotinic acid; pyridoxine HC1; thiamine HC1; sucrose; kinetin; Na 2 EDTA, 2H 2 O; naphthalene acetic acid and mixtures thereof. The culture is carried out at a temperature of 24 to 30 ° C, for a partial pressure of O 2 of about 10%. The culture is done in batches for 7 to 10 days. It can be carried out in batch fed fermentation or continuously.

After culturing in a suitable medium, the dedifferentiated plant cells of the invention are harvested by filtration. The dedifferentiated plant cells thus harvested can then be contacted with a metal ion chelating agent. At this stage, the dedifferentiated plant cells may undergo, prior to any subsequent treatment step, a freezing step followed by a defrosting step. Defrosted defrosted plant cells can then be contacted with a metal ion chelating agent. The dedifferentiated plant cells can then be lyophilized or subjected to an extraction process, these methods being well known to those skilled in the art.

According to the present invention, fresh dedifferentiated plant cells, or freeze-dried cells, or their extracts, or formulated in stable compositions, ie in the presence of a metal ion chelating agent, are used. The freeze-dried dedifferentiated plant cells may be re-extracted in a mixture comprising a metal ion chelator. The lyophilization of the dedifferentiated plant cells advantageously allows the active ingredients to be separated from the biopolymer charges, such as the cell wall polysaccharides. The dedifferentiated cells may be lyophilized and then resuspended in a mixture comprising a metal ion chelator. Cell debris likely to be present in the suspension obtained are removed, for example by passing through a syringe and centrifugation. The suspension freed from cellular debris may be used as an asset derived from dedifferentiated plant cells according to the invention. Any extraction method known to those skilled in the art can be used to prepare a dedifferentiated plant cell extract according to the invention. An "extract" according to the present invention refers to all or part of a fraction comprising the cellular content of dedifferentiated cells of a plant of the species Rosa sp. The cellular content includes in a non-limiting manner the membrane fraction, the cytoplasmic fraction, the cytosolic fraction, the organelles such as mitochondria, chloroplasts, protoplasts and their mixture. An extract may be total, that is to say it contains all the cell fractions, and may be obtained by any method well known to those skilled in the art, for example, by grinding the vegetable matter by means of beads or grains of sand, freezing / thawing, sonication, French press, enzymatic or chemical lysis of the plant wall, etc.

As an extract that is suitable for the invention, mention may be made of aqueous, alcoholic or organic solvent extracts, advantageously an alcoholic organic solvent. By aqueous solvent is meant any solvent constituted totally or by water.

We can thus mention the water itself, hydro-alcoholic solvents in any proportion or the solvents consisting of water and a compound such as propylene glycol in any proportion. Among the alcoholic solvents that may be mentioned include methanol, ethanol, propanol, propanediol, butanol, butanediol. An extraction method that is suitable for the invention may comprise a first step of grinding the dedifferentiated plant cells, for example in a cold aqueous solution, and then a second step of removing particles in suspension from the aqueous solution resulting from the first step, and a third sterilization step of the aqueous solution from the second step. This aqueous solution corresponds to an extract. The extract obtained can then be lyophilized by any conventional freeze-drying method. A powder is thus obtained which can be used directly or mixed in a suitable solvent before use. The dedifferentiated plant cells of the invention may also be used in the form of a lyophilizate. Such lyophilizate can be obtained by any method of lyophilization known to those skilled in the art.

It is understood that these different treatment methods involve the presence of a metal ion chelator, in particular EDTA or a salt thereof, as a stabilizing agent for a solution comprising dedifferentiated cells of a plant of the species Rosa sp.

Uses and Methods The present invention relates, in a first aspect, to the use of a metal ion chelating agent as a stabilizing agent in a composition comprising dedifferentiated cells of a plant of the species Rosa sp. or comprising an extract or lyophilizate thereof.

According to a preferred embodiment of the invention, the method for stabilizing a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof, comprises at least one step contacting said dedifferentiated cells, said extract or said lyophilizate thereof with a metal ion chelating agent.

This contacting of the dedifferentiated plant cells with the metal ion chelating agent is carried out at any stage of a process for obtaining said composition, said extract or lyophilizate. In some embodiments, the contacting of the metal ion chelating agent with the dedifferentiated plant cells is performed at the final stage of a process for obtaining an unstabilized dedifferentiated cell composition of the invention. species Rosa sp., or an extract or lyophilizate thereof. By way of illustration, the metal ion chelating agent may be added to the final unstabilized dedifferentiated cell composition of a plant of the species Rosa sp., Or to an extract or lyophilizate thereof, in order to have a stable or stabilized composition as defined in the present invention. In some embodiments, the metal ion chelating agent is contacted with the dedifferentiated plant cells of Rosa sp. at a plurality of steps, among the steps included in the process for obtaining the composition of said dedifferentiated cells, or an extract or lyophilizate thereof.

In certain embodiments, the method comprises at least the following steps: a) providing a composition, in liquid form, comprising dedifferentiated plant cells of a plant of the species Rosa sp, or comprising an extract or lyophilizate of those and b) adding a metal ion chelating agent to the liquid form composition provided in step a). In general, the composition in liquid form can be chosen from a suspension and a solution. In a preferred embodiment, dedifferentiated plant cells of a plant of the species Rosa sp., An extract or lyophilizate thereof may be in the form of a suspension. In another preferred embodiment, an extract or lyophilizate of dedifferentiated plant cells of a plant of the species Rosa sp. can be implemented in the form of a solution.

Advantageously, the metal ion chelating agent comprises a mono-, di-, tri- or tetra-amine group, to which is covalently attached one or more group (s) derived (s) of the acid acetic.

Thus, advantageously, the chelating agent is chosen from the compounds of general formula (I): NRN - ((CH 2) 2 -NRR) x (formula I), for which x is equal to 1, 2, 3, 4; and for which R is advantageously CH 2 -COOR ', for which R' is chosen from H and Na. In some embodiments, the metal ion chelating agent, for carrying out the uses or processes according to the present invention, is selected from EDTA (ethylenediamine tetraacetic acid), DTPA (diethylene triamine penta acetic acid), MGDA (diacetic methylglycine acid), GLDA (diacetic glutamic acid), NTA (nitrilotriacetic acid), TTHA (triethylene tetraamine hexaacetic acid), EDDS (ethylenediamine disuccinic acid) and their salts. In an advantageous embodiment, the metal ion chelating agent is a sodium salt. In some embodiments, the metal ion chelating agent is an EDTA derivative, for example an EDTA derivative selected from the disodium salt of EDTA and a tetrasodium salt of EDTA (e.g. tetrasodium salt EDTA marketed under the designation Versene 220 Crystals Chelating Agent® by Dow Chemicals). In certain embodiments, the metal ion-chelating agent is selected from the salt of tetrasodium glutamate diacetate (C9H13NO8, for example sold under the designation Dissolvine GL-47® by the company Akzo Nobel), the trisodium ethylenediamine disuccinate compound. For example, in a preferred embodiment, the metal ion chelating agent is used at a concentration of 0.01% w / w (C 10 H 13 N 2 O 8, for example sold under the designation Natrlquest E30® by the company Innospecactive Chemicals). (weight / weight) at 10% w / w, preferably at a concentration ranging from 0.1% w / w to 5% w / w, based on the total weight of said composition. Preferably, the metal ion chelating agent is EDTA, or a salt thereof, and is used at a concentration ranging from 0.01% w / w (w / w) to 10% w / w, preferably at a concentration ranging from 0.1% w / w to 5% w / w, relative to the total weight of said composition. Compositions The present invention relates to a composition, especially a composition for topical use comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof, said composition comprising an agent metal ion chelator. Such a composition is preferably a cosmetic composition, for example a cosmetic composition for topical use. The present invention also relates to a stable dedifferentiated cell composition of a plant of the species Rosa sp., An extract or lyophilizate thereof, comprising, as stabilizing agent, an agent metal ion chelator. Advantageously, the metal ion chelating agent is chosen from EDTA (ethylenediamine tetraacetic acid), DTPA (diethylene triamine penta acetic acid), MGDA (methylglycine diacetic acid), GLDA (diacetic glutamic acid), NTA (nitrilotriacetic acid), TTHA (triethylene tetraamine hexaacetic acid), EDDS (ethylenediamine disuccinic acid) and their salts. According to a particular embodiment of the invention, the metal ion chelating agent may be EDTA or a salt thereof. Advantageously, the chelating agent, and in particular EDTA, or a salt thereof, may be present at a concentration ranging from 0.01% w / w (w / w) to 10% w / w, preferably at a concentration ranging from 0.1% w / w to 5% w / w, relative to the total weight of said composition. Dedifferentiated plant cells of the invention, a lyophilizate or an extract thereof, may be formulated in any compositions suitable for cosmetic or pharmaceutical use. A composition according to the invention comprises a physiologically or pharmaceutically acceptable medium. After culturing in a suitable medium, the dedifferentiated plant cells of the invention are harvested by filtration and can be lyophilized or extracted or dissolved. Advantageously, the dedifferentiated plant cells of the invention, an extract or a lyophilizate thereof can be formulated or put (es) in solution in water or a water-soluble organic solvent, or a mixture of those -this. A water-soluble organic solvent suitable for the invention may be chosen from C 2 to C 6 and preferably C 3 to C 6 hydrocarbon compounds, and comprising from 2 to 6 hydroxyl groups, preferably from 3 to 5 hydroxyl groups, and mixtures thereof. . Among the water-soluble organic solvents that are suitable for the invention, mention may be made in particular of glycols having from 2 to 8 carbon atoms, such as ethylene glycol, propylene glycol or 1,3-propanediol, 1,3-butylene glycol, dipropylene glycol, glycerol, sorbitol, and mixtures thereof. Propylene glycol or 1,3-propanediol is particularly suitable for the invention. A water-soluble organic solvent can constitute from 20 to 99.9% by weight of the composition containing it, preferably from 30 to 90%, preferably from 40 to 80%, and more preferably from 50 to 70% by weight of the composition containing it. A water suitable for the invention may be a thermal and / or mineral water, in particular chosen from Vittel water, the waters of the Vichy basin and the Roche Posay water. The water may constitute from 20 to 99.9% by weight of the composition containing it, preferably from 30 to 90%, preferably from 40 to 80%, and more preferably from 50 to 70% by weight of the composition containing it. Advantageously, the water constitutes up to 50% by weight of the composition containing it. According to a preferred embodiment, a preparation comprising dedifferentiated plant cells of the invention, or comprising an extract or lyophilizate thereof, may comprise at least one hydrophilic gelling agent. Such an agent makes it possible to avoid or reduce the settling of the cells. A hydrophilic gelling agent that is suitable for the invention may be chosen from polysaccharide polymers, and mixtures thereof. For the purposes of the present invention, the term "polysaccharide polymer" means any carbohydrate macromolecule formed by the sequence of a large number of elemental sugars, such as, for example, xylose, glucose, galactose, rhamnose, mannose, fucose, arabinose and their respective acids. The polysaccharide polymers particularly suitable for the present invention are preferably chosen from polymers with molecular weights ranging from 10 to 250 kDa. In particular, polysaccharide polymers suitable for the invention may be cited as pectin; guar gum; cellulose; dextrin; maltodextrin; starch; the gum of Tara; the carob gum; inulin; acacia gum; gum arabic; fucose-rich polymers, such as Fucogel; carrageenans; Konjac gum; xanthan gum; dextran; chitosan; gum tragacanth; ghatti gum; Karaya gum; tamarind gum; agar-agar; alginate; Gellane's gum; and their mixtures. Xanthan gum, locust bean gum, guar gum, gellan gum, agar-agar, alginate, and mixtures thereof are preferably used.

According to a preferred embodiment, a preparation comprising dedifferentiated plant cells of the invention, or comprising an extract or lyophilizate thereof, may comprise a xanthan gum. A gelling agent may be present in a proportion of from 0.01% to 10% by weight, and preferably from 0.1% to 5% by weight of polysaccharide polymer (s), relative to the total weight of the composition . According to another preferred embodiment, a preparation comprising dedifferentiated plant cells of the invention, or comprising an extract or lyophilizate thereof, may comprise at least one sugar.

A sugar suitable for the invention may be a C4 to C6 monosaccharide, and may especially be chosen from inositol, mannitol, glucose, sucrose, trehalose, maltose, xylitol and fructose, and mixtures thereof. . The sugar content in a composition of the invention may vary from 1 to 80% by weight, for example from 5 to 75% by weight, in particular from 10 to 70% by weight, relative to the total weight of the composition. According to a preferred embodiment, the dedifferentiated plant cells of the invention, an extract or a lyophilizate thereof can be formulated in the presence of water, 1,3-propanediol, a chelating agent. metal ions, and / or xanthan gum. The dedifferentiated plant cells of the invention, an extract or a lyophilizate thereof, may be used in an amount representing from 0.001% to 50% by weight of dry matter relative to the total weight of the composition. containing them, and preferably in an amount representing from 0.01% to 30% by weight of dry matter relative to the total weight of the composition. In a preferred embodiment, the dedifferentiated plant cells of the invention, an extract or lyophilizate thereof, may be used in an amount of from 0.1% to 30% by weight of material. dry relative to the total weight of the composition containing them, and advantageously in an amount representing from 0.1% to 20% by weight of dry matter relative to the total weight of the composition. Dedifferentiated plant cells of Rosa sp. of the invention, a lyophilizate or an extract thereof, can be formulated in any cosmetic composition, in particular intended to be ingested, injected or applied to the skin, hair, nails or mucous membranes (oral) , jugal, gingival, genital, conjunctival). Depending on the method of administration chosen, a composition of the invention may be in any of the galenical forms normally used.

For a topical application to the skin, a composition may have the form in particular of aqueous or oily solution or of dispersion of the lotion or serum type, of emulsions of liquid or semi-liquid consistency of the milk type, obtained by dispersion of a phase fat in an aqueous phase (oil-in-water or O / W emulsion) or conversely (water-in-oil or W / O emulsion), or suspensions or emulsions of soft consistency of the cream or gel type, aqueous or anhydrous, or microcapsules or microparticles, or vesicular dispersions of ionic and / or nonionic type. These compositions are prepared according to the usual methods. These compositions may also consist of solid preparations constituting soaps or cleaning bars or may be packaged in the form of an aerosol composition also comprising a propellant under pressure. A composition intended to be applied to the hair may be in the form of aqueous, alcoholic or aqueous-alcoholic solutions, or in the form of creams, gels, emulsions, foams or in the form of aerosol compositions also comprising a propellant under pressure. Such a composition may be presented in the form of a composition for the hair care, such as a shampoo, a setting lotion, a treatment lotion, a cream or a styling gel, a dye composition (especially dyes). oxidation), a coloring shampoo, a restructuring lotion, a permanent composition (especially a composition for the first time of a perm), a lotion or a fall protection gel. For subcutaneous or intradermal administration, a composition may be in the form of an aqueous, oily or serum lotion. A composition suitable for ingestion may be in the form of capsules, granules, syrups or tablets.

The amounts of the various constituents of the compositions according to the invention are those conventionally used in the fields under consideration. When a composition is an emulsion, the proportion of the fatty phase can range from 5% to 80% by weight, and preferably from 5% to 50% by weight relative to the total weight of the composition. The oils, the waxes, the emulsifiers and the co-emulsifiers used in a composition in emulsion form are chosen from those conventionally used in the cosmetics field. The emulsifier and the co-emulsifier may be present in a 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. composition.

When a composition is an oily solution or gel, the fatty phase may represent more than 90% of the total weight of the composition. In known manner, a cosmetic composition as defined by the present invention may also comprise adjuvants conventionally used in the cosmetics field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic additives, preservatives, antioxidants, solvents, perfumes, fillers, filters, odor absorbers and coloring matter. The amounts of these various adjuvants are those conventionally used in the cosmetics field, and are, for example, between 0.01% and 10% of the total weight of the composition. These adjuvants, depending on their nature, can be introduced into the fatty phase, into the aqueous phase and / or into the lipid spherules. As oils or waxes that can be used in the invention, mention may be made of mineral oils (vaseline oil), vegetable oils (liquid fraction of shea butter, sunflower oil), animal oils (perhydrosqualene), synthetic oils ( Purcellin oil), silicone oils or waxes (cyclomethicone) and fluorinated oils (perfluoropolyethers), bees, carnauba or paraffin waxes. To these oils can be added fatty alcohols and fatty acids (stearic acid). As emulsifiers that may be used in the invention, mention may be made, for example, of glycerol stearate, polysorbate 60 and the PEG-6 / PEG-32 / glycol stearate mixture sold under the name Tefose® 63 by the company Gattefosse. As solvents that can be used in the invention, mention may be made of lower alcohols, in particular ethanol and isopropanol, and propylene glycol. As hydrophilic gelling agents that can be used in the invention, mention may be made of carboxyvinyl polymers (carbomer), acrylic copolymers such as acrylate / alkylacrylate copolymers, polyacrylamides, polysaccharides such as hydroxypropylcellulose, natural gums, preferably xanthan, and clays, and lipophilic gelling agents include modified clays such as bentones, metal salts of fatty acids such as aluminum stearates and hydrophobic silica, ethylcellulose, polyethylene.

Cosmetic Use According to a preferred embodiment, a composition according to the present invention can be used as a cosmetic active agent for the care of aged skin or aged hair.

For example, a composition according to the present invention may be used to treat and / or prevent an aesthetic defect resulting from a defective tissue renewal of the aged skin or aged hair in an individual in need thereof; to strengthen, improve or even restore a youthful appearance of the skin and / or the hair.

This composition can be administered, in particular, orally, subcutaneously, intradermally or topically, and preferably by topical. This composition may be administered daily for example, for example, for example, a single administration per day or administration twice a day, for example once in the morning and once in the evening.

A composition according to the present invention may be formulated, for example, in a form suitable for topical administration, such as in the form of a gel, lotion, cream, mousse. Furthermore, a composition according to the invention may be formulated, for example, in a form suitable for oral administration, for example, in the form of capsules, dragees, emulsions, tablets, capsules or capsules. drinkable ampoule. An effective amount of an active ingredient of the invention may be administered in a single dose per day or in divided doses over the day, for example two to three times daily. A cosmetic composition according to the invention may advantageously be used over a period of time varying from one week to several weeks, or even several months, this period being able to be repeated after periods of non-treatment for several months or even several months. years. For example, the administration of an active ingredient according to the invention may be repeated, for example, 2 to 3 times a day, or more, and generally over an extended period of at least 4 weeks, or even 4 to 15 weeks, with if necessary one or more periods of interruption.

For the purposes of the invention, the term "skin" means the entire cutaneous surface of an individual, including the scalp. For the purposes of the invention, the term "hair" means an assembly comprising the hair shaft and the hair follicle. By "aged skin" is meant a general aesthetic state of the skin resulting from chronological aging and / or photoinduced aging. More particularly, the composition according to the present invention aims to prevent and / or reduce and / or treat cutaneous signs of aging.

"Cutaneous signs of aging" means all changes in the external appearance of the skin due to aging whether of chronological origin and / or photo-induced. By way of example of these modifications considered in the invention, there may be mentioned a less homogeneous and less smooth surface, a thinned epidermis and / or less effective in terms of barrier function (which repairs less quickly following aggression), wrinkles and fine lines, wilted skin, lack of elasticity and / or tone of the skin, thinning of the dermis and / or degradation of collagen fibers, resulting in the appearance of soft, wrinkled skin. It also includes all internal changes in the skin that do not always result in a modified external appearance, such as all internal skin damage, and more particularly the degradation of elastin fibers, or elastic fibers, consecutive to exposure to ultraviolet radiation. In particular, the cutaneous signs of aging targeted by the invention are chosen from thinning of the skin, loss of firmness, loss of elasticity, loss of density, or loss of tone of the skin, dryness of the skin. skin, an alteration of the surface appearance of the skin, an appearance of a marked microrelief of the skin, an appearance of roughness, formation and / or presence of fine lines and / or wrinkles, an alteration of the skin, dullness of the skin, papery appearance of the skin, discolouration of the skin, sagging of the skin, or wilting of the skin.

The alterations of the quality of the hair appear with the age consist in particular of a change of aspect of the fiber (fine hair, dull, without holding, soft), without brilliance, and / or graying, more easily brittle, as well as a loss of capillary density. The present composition thus aims to restore / maintain a good capillary density, and / or to improve the quality of the hair and / or the hair fibers, and in particular the aged hair. More particularly, the dedifferentiated plant cells of the invention, an extract or a lyophilizate thereof, make it possible to prevent a loss of capillary density and to protect, reinforce or improve the quality of the aged hair in order to impart a young and dense appearance. EXAMPLES EXAMPLE 1: Stability of a composition comprising a plant extract of dedifferentiated rose cells in the presence of a metal ion chelator 1) Cultivation of dedifferency plant cells of rose (Lancôme rose) from tissue vegetal. The leaves of a rose bush (Rose Lancôme) were decontaminated by the usual techniques of those skilled in the art, and placed on the following culture medium: MICRO NUTRIENTS mg / 1 NH4NO3 1650 KNO3 1900 CaCl2, 2H2O (CX41) 440 MgSO 4 180.8 KH 2 PO 4 170 MICRO NUTRIENTS mg / 1 MnSO 4, 4H 2 O 22.3 ZnSO 4, 7H 2 O 8.6 KI 0.83 Na 2 MoO 4, 2H 2 O 0.25 CuSO 4, 5H 2 O 0.025 Na 2 EDTA, 2H 2 O 37.3 FeSO 4, 7H 2 O 27 VITAMINS mg / 1 MYO-INOSITOL 100 NICOTINIC ACID 0.5 PYRIDOXIN HCl (B6) 0.5 THIAMINE HCl (B1) 0.1 HORMONES mg / 1 naphthalene acetic acid 10 Kinetin 0.06 CARBON SOURCE mg / 1 Sucrose 30000 hormones used are naturally occurring hormones in plants.

Following successive subcultures, dedifferentiated plant cells cultivable fermentereur and therefore industrializable were obtained. The parameters used for the control of the bioreactor culture are as follows: Temperature: from 24 ° C to 30 ° C, and preferably 26-27 ° C Aeration: 10% oxygen partial pressure (p02), regulated by sterile air entering (VVM: Volume of air / volume of fermenter / Minute) and / or stirring avoiding any stress due to shear forces on the cells. The production by batch culture (batch) lasts from 8 to 14 days. The cells are then harvested by filtration on 80-μm mesh of porosity. 2) Treatment of the different lots of dedifferentiated rose plant cells The dedifferentiated Rose Lancôme cells obtained in part 1) are separated into 2 distinct lots. Lot # 1 ("bl") of dedifferentiated cells is subjected to a freezing step at -20 ° C (on dry ice) followed by a first slow thawing step at 4 ° C overnight (12 hours) then a second defrosting step in a water bath at 3040 ° C for 8 hours. After these steps of freezing / thawing the dedifferentiated plant cells are subjected to a step of re-suspension of the thawed batch in a mixture comprising water, in the presence of 1,3 propane-diol (60% by weight of the mixture) and xanthan gum (1% by weight of the mixture), at a rate of 10% by weight of fresh biomass relative to the total weight of the composition. In the sense of the process, the combination of the steps of freezing and thawing can also be referred to as "lyophilization by freezing then thawing" stage. Lot 2 ("b2") of dedifferentiated cells, not having undergone freezing / thawing, is formulated in a mixture of water, in the presence of 1,3 propane diol (60% by weight of the mixture), xanthan gum (1% by weight of the mixture) and a disodium EDTA salt (0.25% by weight of the mixture), at a rate of 10% by weight of fresh biomass relative to the total weight of the mixture. composition. EDTA is known to be a very powerful chelator of calcium, zinc, magnesium, copper and manganese ions, as well as certain ferric species. 3) Analysis of the Visual Appearance and the Texture of the Solutions Obtained in Batches Nos. 1 and 2 The appearance of the solutions was visually performed (staining) and by optical microscopy (aggregation) on solutions at TO (immediately after re-suspension of the lyophilizate), 1 day (T-1j), 15 days (T-15j), 1 month (T-30j) and 2 months (T-60j) after resuspension of the lyophilisate and storage at room temperature at room temperature. shelter from the light. The storage step was also tested at other temperatures, such as 4 ° C, 40 ° C or 45 ° C.

Optical microscopy observations were made in unpolarized and polarized light (Leica DM LB2, Leica Microsystems, Germany). 4) Results 4.1) Aspects of the solutions comprising dedifferentiated cells of roses at room temperature A composition comprising dedifferentiated cells of roses according to batch No. 1 is, just after its preparation, in the form of a translucent gel in which are observed beige / brown cells in suspension. This composition remains stable until T-15j after its preparation and exhibits substantial yellowing at T-30j and T-60j, when stored at room temperature (see Table 1). On the other hand, in the presence of the disodium salt of EDTA, the composition according to batch No. 2 is in the form of a translucent gel with white cells, or even of off-white color, just after manufacture. The occurrence of yellowing is both less intense and delayed over time, since a slight yellowish discolouration is observed only after T-30j (see Table 1). Table 1: Evaluation of the visual appearance (coloration) of the solutions of batches n ° 1 and n ° 2 at room temperature. Lot No T-Oj T-lj T-15j T-30j T-60j 1 Translucent gel with Idem T-Oj Same T-Oj Jaunit Jaunit beige / brown cells in suspension 2 Translucent gel with Idem T-Oj Idem T- Oj Jaunit very yellow / white cells slightly slightly broken off Suspended observation of the dedifferentiated cell compositions of roses according to batches # 1 and # 2 are free of T-Oj aggregates (see Table 2). Some microaggregates are formed in the dedifferentiated # 2 rose cell pool from the first day of storage (T-1j) at room temperature (see Table 2), however, while the number of aggregates increases significantly in the composition according to batch No. 1 after 15, 30 and 60 days of storage (T-15j, T-30j and T-60j), the composition according to batch No. 2, which contains the metal ion chelating agent Disodium EDTA, does not show any aggregates, even after 60 days of storage at room temperature (T-60j) (see Table 2) Table 2: Evaluation of the texture, by the presence of aggregates, of the solutions of the batches n ° 1 and No. 2 at room temperature Lot No. Aggregates T-Oj T-lj T-15j T-30j T-60j 1 None Some Some Some aggregates aggregates aggregates aggregates 2 None None None None None Optical microscopy observation confirms the observations made nude.In effect, a composition according to lot n 1, that is to say comprising rose dedifferentiated cells having undergone freeze / thaw steps without addition of metal ion chelator, shows the presence of aggregates (Figure 1). Note that these aggregates are visible both by light microscopy unpolarized (Figure 1A) and polarized light (Figure 1B). In contrast, the aggregates are non-existent when the dedifferentiated rose plant cells are suspended in a solution comprising a metal ion chelating agent (disodium EDTA salt, not shown). 4.2) Observations of aggregates in solutions comprising dedifferentiated rose cells at temperatures of 4 ° C., 40 ° C. and 45 ° C. Aggregate formation is observed in batch no. 1, at all temperatures tested, from one day after manufacture (T-1j).

For batch 2, no aggregates were observed for the three storage temperatures tested at the times tested (T-lj and T-15j). 5) Conclusions It thus appears that the disodium EDTA salt is capable of conferring a stability on a composition comprising de-planting rose plant cells. The presence of disodium EDTA in the composition thus improves the homogeneity, preventing the formation of aggregates of plant material, and furthermore makes it possible to maintain the initial white / off-white color of the cells of roses, during the process of preservation or storage of the composition over time. EXAMPLE 2 Formulation Examples 1) Day cream Ingredients% (wt / wt) Sodium hydroxide 0.16995 Adenosine 0.04 Disodium EDTA 0.025 Preservative 0.7 Mineral oil 2 Shea butter (Butyrospermum parkii) 1.5 Extract pink according to batch No. 2 of Example 1 Hydrogenated polyisobutene 3 Xanthan gum 0.2 Sodium polyacrylate 0.6 Dimethicone 4 Water Qs 100 Glycerin 5 Stearic acid 1.5 2) Lotion Ingredients% (wt / wt) Sodium hydroxide 0.114 Disodium EDTA 0.1 Preservative 0.7 Rose extract according to Lot No. 2 of Example 1 1 Xanthan gum 0.02 Water Qs 100 Glycerin 3 Propylene glycol 5 Hydrogenated castor oil PEG-60 0 , 6 Topical application of a day cream or a lotion according to the present invention provides a beneficial effect in the treatment and / or prevention of an aesthetic defect due to a defective tissue renewal of the aged skin or aged hair, and / or in strengthening, has improving or even restoring a young aspect of the skin and / or the hair.

Claims (11)

REVENDICATIONS1. Use of a metal ion chelating agent as a stabilizing agent in a composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof. A method for stabilizing a composition comprising dedifferentiated cells of a plant of the species Rosa sp., An extract or lyophilizate thereof, which method comprises at least one step of contacting said dedifferentiated cells, said extract or said lyophilizate thereof, with a metal ion chelating agent. 3. Process according to claim 2, comprising at least the following steps: a) providing a composition, in liquid or gelled form, of dedifferentiated plant cells of a plant of the Rosa sp species, an extract or a lyophilisate of these, and b) adding a metal ion chelating agent to the composition in liquid or gelled form provided in step a). 4. Use according to claim 1 or method according to one of claims 2 and 3, wherein the metal ion chelating agent is selected from EDTA (ethylenediamine tetraacetic acid), DTPA (diethylene triamine penta acetic acid) , MGDA (diacetic methylglycine acid), GLDA (diacetic glutamic acid), NTA (nitrilotriacetic acid), TTHA (triethylene tetraamine hexaacetic acid), EDDS (ethylenediamine disuccinic acid) and their salts. 5. Use according to claim 1 or method according to one of claims 2 to 4, wherein the metal ion chelating agent is EDTA or a salt thereof. 6. Use according to claim 1 or method according to one of claims 2 to 5, wherein the metal ion chelating agent, preferably EDTA or a salt thereof, is used at a concentration ranging from 0, 01% w / w at 10% w / w, preferably at a concentration ranging from 0.1% w / w to 5% w / w, relative to the total weight of said composition. A composition comprising dedifferentiated cells of a plant of the species Rosa sp., Or comprising an extract or lyophilizate thereof, said composition comprising a metal ion chelating agent. The composition of claim 7, wherein the metal ion chelating agent is selected from EDTA (ethylenediamine tetraacetic acid), DTPA (diethylene triamine penta acetic acid), MGDA (methylglycine diacetic acid), GLDA (Diacetic glutamic acid), NTA (nitrilotriacetic acid), TTHA (triethylene tetraamine hexaacetic acid), EDDS (ethylenediamine disuccinic acid) and their salts. The composition of claim 7, wherein the metal ion chelating agent is EDTA or a salt thereof. 10. Composition according to one of claims 7 to 9, wherein the metal ion chelating agent, preferably EDTA or a salt thereof, is present at a concentration ranging from 0.01% w / w to 10% w / w, preferably at a concentration ranging from 0.1% w / w to 5% w / w, relative to the total weight of said composition. 11. Cosmetic use of a composition according to one of claims 7 to 10 as an active agent for the care of aged skin or aged hair.