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    • 专利摘要:
    According to the invention, it is proposed the use of undifferentiated or dedifferentiated plant cells of Leontopodium alpinium obtained by cell culture in vitro, for a non-therapeutic cosmetic treatment to restore the homeostasis of the cells of the aged skin and to increase their metabolic activity and energy. Thanks to this treatment one obtains a restorative and smoothing effect on an already aged skin, notably pronounced on the drooping surface of the neck.
    公开号:FR3031454A1
    申请号:FR1550239
    申请日:2015-01-13
    公开日:2016-07-15
    发明作者:Olga Gracioso；Caroline Ringenbach；Emmanuel Doridot；Philippe Mondon
    申请人:Sederma SA；
    IPC主号:
    专利说明:

    [0001] The present invention relates to the use of the plant Leontopodium nivale subsp. alpinum Cass, hereinafter called Leontopodium alpinum, for a cosmetic treatment (topically or orally). The present invention thus relates to the industry of cosmetics and personal care and hygiene products, applying to the skin and its integuments (such as hair, eyelashes, eyebrows, nails, hair) of mammals, animals or humans. . Leontopodium alpinium, also known as Edelweiss, is a herbaceous plant of the family Asteraceae. It grows in altitude beyond 1500 m on the mountain ranges of the Pyrenees, the Alps and the Himalayas, in hospitable areas such as ravines, rocky areas, cold and very exposed to UV. As a result, it is a plant that has excellent adaptation to these extreme conditions through a variety of molecules of interest and through protective hairs on its flower and leaves. On the other hand, it is relatively little distributed and thus rather rare. Products derived from the plant can be obtained by conventional extraction methods directly from the whole plant or its parts or by in vitro culture methods either by cell culture or by tissue culture from cell lines. or tissues from different organs of the plant. The present invention relates more particularly to the products obtained following the in vitro culture of cells or tissues. Obtaining by in vitro culture extracts of plant origin has many advantages over the agro-industrial pathway (plant culture in the field and subsequent extraction in the plant). Because of the complete control of the culture conditions, extracts obtained by in vitro culture are free of toxic substances (herbicides, pesticides, fertilizers, heavy metals and other contaminants, such as those that can come from plant pests). Moreover, the strict control of the in vitro culture conditions reduces the risk of spontaneous variation of the strain and guarantees a reproducible profile of secondary metabolites which correspond to the molecules of interest sought, unlike the field crop where the problem of variability, related to climatic, meteorological and geographical conditions and their hazards. Moreover, this technology overcomes obstacles such as the natural biological cycle of the plant and the seasonality of production of secondary metabolites, allowing a better security and rapidity of supply. In addition, the environmental impact is minimal because it substantially limits water consumption, avoids the consumption of arable land, and prevents soil pollution. In addition, the biodiversity is preserved since it is enough of a plant or even of a seed, to initiate a new culture in vitro. Finally, this technology offers the possibility of orienting cellular metabolism towards the production of the molecules of interest (elicitation of cultures) and of carrying out controlled and relatively rapid protocols with a view to increasing the yields of certain molecules, in particular those produced in vitro. low amount in the plant. Among the techniques that exist to date in the field of in vitro culture of plants, it is possible to use according to the invention: the culture of undifferentiated or dedifferentiated cells: this type of method firstly comprises the creation of cell lines strongly proliferating in agar medium either from meristematic cells that are undifferentiated cells, or from dedifferentiated cells (growing in the form of calli, following the removal of a fragment of plant, leaf, stem, root or other). These lines are then cultured in a liquid medium so as to substantially increase the biomass. At the end of the growth cycle and under environmental conditions to be defined and optimized (search for the right elicitation medium), the cell biomass will synthesize the molecules of interest. The culture is then stopped and extracted at the optimum time to obtain a maximum amount of molecules of interest. Originally, one can also use existing cell lines already commercially available. - tissue culture or organ: this type of culture may concern the root part ("root culture" in English), the aerial part ("shoot culture") or somatic embryos ("somatic embryo"). In this type of method, we distinguish between cultures that have undergone genomic transformation by Agrobacterium rhizogens (roots) or Agrobacterium tumefasciens bacteria (stems). The crops thus transformed of roots or aerial parts have a high growth rate and are genetically very stable. They are used to synthesize the molecules of interest after optimization of the elicitation parameters. These cultures are then extracted by conventional routes. Preferably, the present invention is more particularly directed to products derived from the in vitro culture of undifferentiated or dedifferentiated cells, hereinafter referred to as plant cell culture.
    [0002] The in vitro methods of cell culture schematically consist of: - if necessary in a first step, to establish cell lines from calli (clusters of undifferentiated or dedifferentiated cells) obtained on cuts of parts of plants (leaf, root, stem , buds ....); - select a cell line capable of producing a large scale biomass of cells according to pre-established criteria (constant phenotype and optimal and constant production of selected metabolites, ability to proliferate); and then from this selected line, to generate said cellular biomass, optionally with an elicitation step, preferably at the end of the proliferation phase; and thirdly, to treat the cell biomass obtained to recover the whole cells, to break up the cell aggregates by homogenization under high pressure or to lyse these cells and, if appropriate, to extract the contents of said cells.
    [0003] It is these extracts and / or whole or lysed cells which can then be used in cosmetic compositions as an active ingredient with a physiologically acceptable excipient and optionally other complementary active ingredients. Patent Application EP2319914 describes this in vitro technique to obtain undifferentiated cells in culture with a high yield of caffeic acid derivatives for a theoretical list of 33 species of plants including Leontopodium alpinum. The caffeic acid derivatives concerned include phenylpropanoid glycosides as well as caffeoylquinic acids. Inhibitory activity of hyaluronidase is presented for Leontopodium alpinum. Protecting hyaluronic acid from digestion with hyaluronidase makes it possible, as a preventive measure, to preserve the integrity of the dermis. The present invention aims to propose a new use of Leontopodium alpinum cells obtained by an in vitro cell culture method for a cosmetic treatment. For this purpose, it proposes the use of undifferentiated or dedifferentiated Leontopodium alpinium cells obtained by an in vitro cell culture method for a non-therapeutic cosmetic treatment to restore the homeostasis of the aged skin cells and to increase their metabolic activity. energy. The homeostasis of the cells of the elderly dermis results in: 1) a rebalancing between MMP ("Matrix Metallo Proteases": proteases of the dermal matrix) and TIMP ("Tissue Inhibitor of Metallo Proteinases" - tissue inhibitor of the proteases of the matrix 20 dermal), in favor of the latter. MMPs are numerous (more than 20), they are produced in a latent form (pro-MMP) in order to avoid an uncontrolled runaway. Their production increases with age, but acute or chronic stress amplifies this phenomenon. Their excessive production leads to the reduction of the behavior of the dermis, to the loss of its density, to its refinement. Thus the MMP 25 lead to age the skin faster and worse. While the dermal fibers in the young skin are long, intact and dense, those of the aged skin are on the contrary fragmented and less dense. This lower quality of the fibers reduces the dynamic tension of the dermis, the interactions between the MEC and the fibroblasts being reduced sometimes by 80%. MMPs are naturally controlled in tissues by their TIMP inhibitors which avoids blind activity. It is the balance of the TIMPs compared to the MMPs which determines the level of activity of the latter. TIMPs are small glycoproteins whose production is associated with the reduction of chronic pathologies linked to MMPs and a reduction in UV-related photo-damage. By associating with MMPs, TIMPs neutralize them and thus limit the fragmentation of the dermal matrix, which thus retains its elasticity. Furthermore, TIMP-1 is considered to be a favorable survival factor for fibroblasts and keratinocytes.
    [0004] The fall in production of TIMP-1 is observed under the effect of UV irradiation but also during the senescence of fibroblasts. This results in increased activity of MMPs. 2) Stimulation of thrombospondin production (TSP). They are matrix glycoproteins produced by the epidermis and the dermis whose scientific interest is increasing. TSP-1, once overexpressed experimentally in the mouse epidermis effectively reduces the harmful effects of UVB irradiation. Interestingly, there has been a marked improvement in the organization of the collagen and elastin fibers of the underlying dermis as well as the reduction of formation of wrinkles on the skin. This would be related to the control of proangiogenic and protease factors. Its counterpart, the TSP-2, also has a significant potential anti-aging role. Indeed, mice that can not produce this protein, show dermas with collagen fibers less organized, disordered and their skin is more fragile to the extension. Their fibroblasts provide artificial dermas that are also looser and less dense that do not contract normally. TSP-1 and TSP-2 appear to act on several key points of aging, in particular by inhibiting the activity of MMPs, including that of MMP-9. Moreover, the increase in production of TSP-1 is also known to restore the synthesis of pro-collagen decreased by UVB irradiation. This recovery involves the activation of well-known TGF-f3 to stimulate collagen synthesis. In contrast, fibroblasts that can not produce TSP-2 overproduce MMP-9. Thus, it is clear that the expression of TSPs, including by keratinocytes, represents a favorable asset for the quality of the dermis and the skin and, alongside the TIMPs, limit its aging. Increased metabolic activities of dermal fibroblasts result in improved contractile capabilities and increased production of dermal macromolecules. Regarding the energetic activity of fibroblasts: each cell of the body produces, thanks to oxygen from the air, its own energy from production units, called the mitochondria, in greater or lesser numbers depending on the energy requirements of the fabric. With age the mitochondria lose efficiency and convert less oxygen than before. Fibroblasts possess filament-shaped mitochondria of 1 to 10um unlike other cell types, with a high multiplicative power, where they are rather ovoid. However, the two forms, isolated ovoid (fission) or filamentous network (fusion), coexist in each cell type. Preserving the dynamics of passage from one state to another is crucial for the maintenance of mitochondrial functions. If fission is essential for cell division by distributing the pool of mitochondria in the daughter cells, the fusion allows to pool genes and molecules necessary for the maintenance of cellular respiration.
    [0005] It has been observed that the senescent cells are rich in fused form, which again allows an extension of their survival.
    [0006] According to the invention, the undifferentiated or dedifferentiated cells of Leontopodium alpinium can be used after having been mixed under high pressure in order to homogenize the medium and to break up the cell aggregates. According to the invention, the undifferentiated or dedifferentiated cells of Leontopodium alpinium can be used whole or lysed, or else in the form of a cell extract made from these cells. According to the invention, the undifferentiated or dedifferentiated cells of Leontopodium alpinium can be extracted by any physiologically acceptable solvent, or by any mixture of these solvents. The extraction can be carried out according to the various known processes that can be combined: hot, by maceration, decoction, infusion, pressure, leaching, with ultrasound, with microwaves, by lysing the cells by any appropriate chemical or physical process. Phase separation can be by filtration or centrifugation. Alternatively, it is possible to extract the biomass with a supercritical or subcritical fluid. According to the invention, it is also possible to envisage a thorough purification of the cell extract by all commercially available methods, by liquid-liquid partitioning or chromatography, in particular by means of an adsorbent resin, in order to concentrate the molecules. Of interest, such as the two lipopic acids A and / or B. According to the invention, the undifferentiated or dedifferentiated cells of Leontopodium alpinium can also be used in a spray-dried or preferably lyophilized form. This allows their long-term storage and preserves their biological activity. In the remainder of the description, the expression "plant cells" will encompass the undifferentiated or dedifferentiated Leontopodium alpinium cells obtained by an in vitro cell culture process, whether they are whole or lysed, whether or not they have been homogenized. at high pressure, whether fresh or dry, as well as extracts from these cells.
    [0007] According to the invention, the plant cells may be incorporated (suspended or solubilized) in a physiologically acceptable medium and used to produce a cosmetic composition intended to restore the homeostasis of the aged skin cells and to increase their metabolic and energetic activity. . Preferably, according to the invention, the physiologically acceptable medium is a hydrophilic matrix.
    [0008] More preferably, the cosmetic treatment according to the invention is topical. Surprisingly, the Applicant has shown by in vivo tests that the cosmetic treatment of aged skins by the application of a composition containing undifferentiated or dedifferentiated Leontopodium alpinium cells obtained by an in vitro cell culture method induced on these skins a curative or restorative effect, tensor and / or smoothing, and in particular on the drooping surface of the neck and the fold of the valley of tears.
    [0009] The results of these tests are presented in more detail below in the description. The invention finally proposes the use of plant cells, as defined above, for the manufacture of a non-therapeutic cosmetic active ingredient, as well as the use of a composition comprising said suspended active ingredient and / or solubilized in a physiologically acceptable medium, which may be a hydrophilic matrix, to restore the homeostasis of the aged skin cells and increase their metabolic and energetic activity. According to the invention, plant cells rich in leontopodic acids A and / or B, preferably A and B, are used. Proteins, amino acids, phytosterols, lipids and polysaccharides have also been identified as categories of compounds in the plant cells used according to the invention. According to other features, the present invention therefore proposes a cosmetic active ingredient, for a use according to the invention, comprising undifferentiated or dedifferentiated plant cells of Leontopodium alpinium obtained by an in vitro cell culture process and comprising at least 0, 04% of lactic acid A and B in a physiologically acceptable matrix. Preferably, in this ingredient and as developed above, the plant cells are whole and / or lysed, or the cells are in the form of a cell extract of said whole cells and / or lysed. In order to obtain the dedifferentiated or undifferentiated cells which can be used according to the invention, the following process can be used: 1) from a selected line of Leontopodium alpinum, to produce a critical pre-biomass by successive pre-cultures and to growing sizes; 2) producing a biomass of said undifferentiated cells in a bioreactor from said pre-biomass and a suitable culture medium; and 3) separating said biomass enriched in leontopodic acids from said culture medium and thereby recovering said undifferentiated cells. According to other optional features: 1) the bioreactor production step may comprise an elicitation step, which advantageously makes it possible to increase the levels of the A and / or B luteopodic acids or to vary the relative proportion thereof; and / or 2) the biomass from the reactor is collected by filtration after a culture time between 7 and 21 days; preferably between 10 and 14 days, which advantageously makes it possible to produce the highest amount of biomass, with a high viability; and / or 3) the biomass can be subjected to a homogenization step under high pressure, in order to break the cell aggregates; and / or 3031454 7 4) an additional step of drying the cell biomass may be added, so as to preserve it in the long term; and / or 5) The cells may be extracted in particular for the purpose of enrichment of leprosyic acids A and B.
    [0010] In general, the elicitation of the compounds of interest can be done by the addition to the culture of microbial fractions (especially saccharomyces yeasts): the addition to the culture of molecules of biological origin, for example chitosan, methyljasmonate, jasmonic acid, salicylic acid; the addition to the culture of molecules of non-biological origin, for example paclobutrazol; the application to culture of a change in temperature, pH or osmotic stress induced by a non-metabolizable sugar, such as for example mannitol; the use of an even more drastic impoverishment of the environment in macroelements and sugar; the addition to the culture of adsorbent resins which in addition to elicit the production of the compounds of interest can trap them. Preferably according to the invention, the elicitation is carried out by modifying the culture medium, especially the nutrient levels.
    [0011] Preparation of compositions for carrying out the invention A cosmetic composition, in particular a topical composition, comprises plant cells in a physiologically acceptable medium. Depending on the excipient and the dosage in plant cells, this composition will constitute a concentrated active ingredient or a less concentrated final composition directly intended for the end user.
    [0012] By "physiologically acceptable medium" is meant according to the present invention, without being limiting, an aqueous or hydroalcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion, an aqueous gel, a gel anhydrous, a serum, a dispersion of vesicles, a powder. "Physiologically acceptable" means that the compositions are suitable for topical or oral use, in contact with mucous membranes, nails, scalp, hair, hair and mammalian skin and more particularly human skin, compositions be ingested or injected into the skin, without risk of toxicity, incompatibility, instability, allergic response, and others. This "physiologically acceptable medium" forms what is conventionally called the excipient of the composition. The plant cells may be combined with other active ingredients at effective concentrations that can act synergistically or in a reinforcing manner to achieve the desired effects described for the invention, such as the following: radiation filtering agents, in particular UVA and / or or UVB, moisturizing, humectant, calming, myorelaxant, slimming, restructuring, firming, plumping, tensor, acting on the microcirculation, acting on inflammation, free radicals, anti-wrinkles, lightening, acting on radiance, anti -glycation, pro-pigmenting, acting on the stratum corneum, on the dermal-epidermis junction, on the production of protein HSPs, on firmness, elasticity, skin tone, regrowth of peptide hairs, vitamins etc. The plant cells may be applied according to the invention to the face, body, décolleté, scalp, hair, eyelashes, hairs, in any form or vehicle known to the man of the art, especially in the form of solution, dispersion, emulsion, paste or powder, individually or premixed or be conveyed individually or pre-mixed by vectors such as macrocapsules, microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macro-sponges, micro-sponges or nanoepongs, microemulsions or nanoemulsions, or adsorbed onto powdery organic polymers, talcs, bentonites, spores or exines and other mineral or organic substrates. In cosmetics in particular, applications can be proposed especially in the skincare ranges of the face, body, hair and body hair and makeup-care ranges, including eyelashes and eyebrows.
    [0013] In general, the plant cells according to the present invention may be used in any form, in a bound form, incorporated or adsorbed on macro-, micro-, and nanoparticles, or on macro-, micro-, and nanocapsules, for the treatment of textiles, natural or synthetic fibers, wools, and any materials intended to come into contact with the skin and which may be used in clothing, underwear, daytime or nightwear, tissues, or tissues, to exert its cosmetic effect through this skin / textile contact and allow continuous topical delivery. The CTFA ("International cosmetic ingredient dictionary & handbook" (13th edition, 2010) published by "The Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, DC) describes a wide variety, without limitation, of cosmetic ingredients and Pharmaceuticals commonly used in the skincare industry, which are suitable for use as additional ingredients in the compositions of the present invention. Other additional skin care actives that are particularly useful can be found in Sederma's commercial literature and at www.sederma.com. The following commercial active agents may also be mentioned as examples: betaine, glycerol, Actimoist Bio 2TM (Active Organics), AquaCacteenTM (Mibelle AG Cosmetics), AquaphylineTM (Silab), AquaregulKTM (Solabia), CarcilineTM ( Greentech), CodiavelaneTM (Biotech Marine), DermafluxTM (Arch Chemicals, Inc.), Hydra'FlowTM (Sochibo), Hydromoist LTM (Symrise), RenovHyalTM (Soliance), SeamossTM (Biotech Marine), ArgirelineTM (trade name acetyl hexapeptide) Lipotec-3), spilanthol or an Acmella oleracea extract known as Gatuline ExpressionTM, an extract of Boswellia serrata known as BoswellinTM, Deepaline PVBTM (Seppic), Syn-AKETM 3031454 (Pentapharm), AmelioxTM , BioxiliftTM (Silab), PhytoCellTecTMArgan (Mibelle), Papilactyl DTM (Silab), PreventheliaTM (Lipotec), SubliskinTM (Sederma), VenuceaneTM (Sederma), Moist 24TM (Sederma), Vegesome Moist 24TM (Sederma), EssenskinTM (Sederma), JuvinityTM (Sederma), RevidratTM (Sederma), ResistemTM (Sed erma), ChronodynTM (Sederma), KombuchkaTM (Sederma), ChromocareTM (Sederma), CalmosensineTM (Sederma), Glycokin factor STM (Sederma), BiobustylTM (Sederma), IdealiftTM (Sederma), Ceramide 2TM, Ceramide A2TM and Ceramide HO3TM ( Sederma), LeganceTM (Sederma), IntenslimTM (Sederma), ProdiziaTM (Sederma), SenestemTM (Sederma), BeautifeyeTM (Sederma), PacifeelTM (Sederma), SebulessTM (Sederma), NG Unsaponifiables of Shea ButterTM (Sederma) or blends of these.
    [0014] Among the plant extracts which can be combined with the plant cells according to the invention, ivy extracts, for example climbing ivy (Hedera Helix), Bupleurum chinensis, Bupleurum Falcatum, can also be mentioned in particular. arnica (Arnica Montana L), rosemary (Rosmarinus officinalis / V), marigold (Calendula officinalis), sage (Salvia officinalis L), ginseng (Panax ginseng), ginko biloba, St. John's wort (Hyperycum Perforatum), frog (Ruscus 15 aculeatus L), water-mantle (Filipendula ulmaria L), orthosiphon (Orthosiphon Stamincus Benth), seaweed (Fucus Vesiculosus), birch (Betula alba), green tea, kola nut ( Cola Nipida), horse chestnut, bamboo, Centella asiatica, heather, fucus, willow, pilosella, escin extracts, cangzhu extracts, chrysanthellum indicum extracts, plants of the genus Armeniacea, Atractylodis Platicodon , Sinnomenum, Pharbitidis, Flemingia, Coleus as C.
    [0015] Forskohlii, C. blumei, C. esquirolii, C. scutellaroides, C. xanthantus and C. barbatus, as a raciness extract of Coleus barbatus, extracts of Ballot, Guioa, Davallia, Terminalia, Barringtonia, Trema, antirobia, cecropia , argania, dioscoreae as Dioscorea opposita or mexico, extracts of Ammi visnaga, Siegesbeckia, in particular Siegesbeckia orientalis, plant extracts of the family Ericaceae, in particular extracts of blueberries (Vaccinium angustifollium) from Arctostaphylos uva ursi, alpe vexa, plants containing sterols (in particular phytosterols), Manjistha (extract of plants of the genus Rubia, in particular Rubia Cordifolia), Guggal (extract of plants of the genus Commiphora, in particular Commiphora Mukul), a kola extract, chamomile, violet clover, Piper methysticum (Sederma Kava Kava), Bacopa monieri (BacocalmineTM Sederma) and sea whip, Glycyrrhiza glatira, mulberry, tea tree, Larrea 30 divaricata, Rabdosia rubescens, Euglena gracilis, Fibraurea recisa Hirudinea, Chaparral Sorghum, sunflower, Enantia chlorantha, Mitracarpe of the genus Spermacocea, Buchu barosma, Lawsonia inermis L., Adiantium Capillus-Veneris L., Chelidonium malus, Luffa cylindrical, Japanese Mandarin (Citrus reticulata Blanco var. unshiu), Camelia sinensis, Imperata cylindrical, Glaucium Flavum, Cupressus Sempervirens, Polygonatum multiflorum, 35 loveyly hemsleya, Sambucus Nigra, Phaseolus lunatus, Centaurium, Macrocystis Pyrifera, Turnera Diffusa, Anemarrhena asphodeloides , Portulaca pilosa, Humulus 3031454 10 lupulus, Arabica coffee, Ilex Paraguariensis, Globularia Cordifolia, Oxydendron arboreum, Albizzia julibrissin and Zingiber zerumbet smith. The compositions according to the present invention may comprise one or more peptides, including, but not limited to, di-, tri-, tetra-, penta- and hexapeptides and derivatives thereof. According to a particular embodiment, the concentration of the additional peptide in the composition varies between 1 × 10 -7% and 20%, preferably between 1 × 10 -6% and 10%, preferably between 1 × 10 -5% and 5%, weight. The term "peptide" herein refers to peptides containing 10 amino acids or less, their derivatives, isomers and complexes with other species such as a metal ion (e.g. copper, zinc, manganese, magnesium, and others). The term "peptides" refers to both natural peptides and synthetic peptides. It also refers to compositions which contain peptides and which are found in nature, and / or which are commercially available. Nonlimiting examples of dipeptides that may be used in the context of the present invention include Carnosine (beta-AH), YR, VW, NF, DF, KT, KC, CK, KP, KK or TT. Non-limiting examples of tripeptides include RKR, HGG, GKH, GGH, GHG, KFK, KPK, KMOK, KMO2K or KAvaK. Non-limiting examples of tetrapeptide are RSRK (SEQ ID NO: 1), GQPR (SEQ ID NO: 2) or KTFK (SEQ ID NO: 3). A non-limiting example of pentapeptide is KTTKS (SEQ ID NO: 4) and hexapeptides GKTTKS (SEQ ID NO: 5) and VGVAPG (SEQ ID NO: 6). Other peptides which may be used in the context of the present invention may be chosen from, without this list being limiting: the lipophilic derivatives of peptides, preferably the palmitoyl derivatives, and the complexes with the metal ions mentioned above (eg complex tripeptide copper HGG). Preferred dipeptides include, for example, N-Palmitoyl-beta-Ala-His, N-Acetyl-Tyr-Arghexadecylester (Calmosensine ™, Idealift ™, Sederma), Pal-KT, Pal-RT (Sederma). The preferred tripeptides include N-Pal-Gly-Lys-His, (Pal-GKH, Sederma), the copper derivative of HGG (LaminTM, Sigma), lipospondin (N-Elaidoyl-KFK) and its substitution analogues. conservative, N-Acetyl-RKR-NH2 (Peptide CK +), N-Biot-GHK (Sederma), Pal-KMO2K (Sederma) and their derivatives. Tetrapeptide derivatives which may be used in the context of the present invention include, but are not limited to, N-Pal-GQPR (Sederma) (SEQ ID NO: 7), Ela-KTFK (SEQ ID NO: 8). Useful pentapeptide derivatives are, but are not limited to, N-Pal-KTTKS (SEQ ID NO: 9) (MATRIXYL ™, Sederma), N-Pal-Tyr-Gly-Gly-Phe-X (SEQ ID NO: 10). ) with X being Met or Leu or their mixture. Useful hexapeptide derivatives include, but are not limited to: N-Pal-VGVAPG (SEQ ID NO: 11), Pal-GKTTKS (SEQ ID NO: 12), and derivatives. We can also mention the mixture Pal-GHK and Pal-GQPR (SEQ ID NO: 7) (MatrixylTm 3000, Sederma). Preferred commercially available compositions containing a tripeptide or derivative include Biopeptide-CLTM, MaxilipTM or Sederma BiobustylTM. Preferred, commercially available sources of tetrapeptide sources include: RIGINTM, Eyeliss ™ Matrixyl ™ Reloaded and Matrixyl 3000 ™, which contain between 50 and 500 ppm of palmitoyl-GQPR (SEQ ID NO: 7) and an excipient, proposed by Sederma. The following commercial peptides can also be mentioned as additional active ingredients: Vialox ™ (INCI name = Pentapeptide-3 (synthetic peptide including alanine, arginine, isoleucine, glycine and proline)), Syn-ake ™ ((3-Ala) Pro-Dab-NH-Bz1) or Syn-Coll ™ (Pal-LysVal-Lys-OH) sold by Pentapharm, - Argireline ™ (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH 2 (Nom INCI = Acetyl hexapeptide-3) (SEQ ID NO: 13), Leuphasyl ™ (Tyr-D-Ala-Gly-Phe-Leu) (SEQ ID NO: 14), Aldenine ™ (Gly-His-Lys), TrylagenTM (INCI name = Pseudoalteromonas Ferment Extract, Hydro lyzed Wheat Protein, Hydro Lyzed Soy Protein, Tripeptide-10 Citrulline (product of the reaction of Citrulline and Tripeptide-10 (synthetic peptide consisting of aspartic acid, isoleucine and lysine)), Tripeptide-1), EyeserylTM (Ac- (3-Ala-His-Ser-His) (SEQ ID NO: 15), SerilesineTM (Ser-IleLys-Val-Ala-Val) (SEQ ID No. 16) or DecorinylTM (INCI name: Tripeptide-10 Citrulline = 1 5 product of the reaction of Citrulline and Tripeptide-10 (synthetic peptide consisting of aspartic acid, isoleucine and lysine) sold by Lipotec, - CollaxylTM (Gly-Pro-Gln-Gly-Pro-Gln ( SEQ ID No. 17)) or QuintescineTM (Cys-Gly) sold by Vincience, - CytokinolTMLS (casein hydrolyzate) sold by Serobiological Laboratories / Cognis, 20 - KollarenTM (Gly-His-Lys), IP2000TM (Pal-Val-Tyr-Val) or MelipreneTM (INCI name = Monofluoroheptapeptide-1: product of the reaction of acetic acid and a synthetic peptide containing arginine, glycine, glutamic acid, histidine, norleucine, p- fluorophenylalanine and tryptophan) sold by the European Institute of Cell Biology, - NeutrazenTM (Pal-His-D-Phe-Arg-NH2) sold by the company Innovations, or 25 - the BONT-L-PeptideTM (INCI name = Palmitoyl Hexapeptide-19: product of the reaction of palmitic acid and Hexapeptide-19 (synthetic peptide consisting of asparagine, Aspartic acid, lysine and methionine), Timp-Peptide ™ (INCI name = Acetyl Hexapeptide-20: product obtained by acetylation of Hexapeptide-20 (synthetic peptide consisting of alanine, glycine, lysine, valine and proline) or ECM Modulin TM (INCI name = Palmitoyl Tripeptide-28: product of the reaction of palmitic acid and Tripeptide-28 (synthetic peptide consisting of arginine, lysine and phenylalanine) sold by the company Infinite Activos. More specifically, according to the invention the plant cells can be combined with at least one of the compounds chosen from vitamin B3 compounds, compounds such as niacinamide or tocopherol, retinoid compounds such as retinol, hexamidine, α-lipoic acid, resveratrol or DHEA, peptides, especially N-acetyl-Tyr-Arg-O-hexadecyl, Pal-VGVAPG (SEQ ID NO: 11), Pal-KTTKS (SEQ ID NO: 9), Pal-GHK, Pal-KMO2K and Pal-GQPR (SEQ ID NO: 7), which are conventional active ingredients used in topical cosmetic or dermopharmaceutical compositions.
    [0016] The present invention also provides a topical cosmetic or dermatological treatment method for improving the appearance and general condition of the skin and its integuments, including topical application to the skin of a subject who in need of an effective amount of plant cells or a composition comprising them, in a physiologically acceptable excipient.
    [0017] By "topical treatment" or "topical use" is meant an application that is intended to act at the place where it is applied: skin, mucous membrane, superficial body growths. The composition comprising the plant cells according to the invention may be applied locally to the targeted zones. The "effective" amount depends on a variety of factors, such as age, condition of the patient, severity of the disorder or condition, and mode of administration. An effective amount means a non-toxic amount sufficient to achieve the desired effect. According to other features of the invention: the physiologically acceptable medium is a hydrophilic matrix in which said plant cells are suspended and / or solubilized; and / or the active ingredient comprises a thickener; and / or - the active ingredient comprises at least 0.04% of lontopodic acids A and B relative to the total weight of the ingredient. This ingredient can then be used in cosmetic formulations between 0.1 and 10%, preferably between 1 and 5%, more preferably between 2 and 3% and generally 2% by weight of said formulation; which corresponds to levels of lontopodic acids A and B in the cosmetic formulations of between 0.00004% and 0.004%, preferably between 0.0004% and 0.002%, more preferably between 0.0008% and 0.0008%. , 0012% and generally at least 0.0008% relative to the total weight of the composition. All percentages and ratios used in this application are by weight of the total composition and all measurements are made at 25 ° C unless otherwise specified.
    [0018] By way of example, for a cosmetic facial treatment, the European Cosmetics Directive has fixed a standard application amount of a cream of 2.72 mg / cm 2 / day / person and for a body lotion 0.5 mg / cm2 / day / person. According to other particularities, the cosmetic treatment method according to the invention may be associated with one or more other treatment methods aimed at the skin, such as, for example, treatments by light therapy, by heat or by aromatherapy.
    [0019] According to the invention, it is possible to propose devices with several compartments or kits intended for the implementation of the method described above, and which could include, by way of example, and without being limiting, in a first compartment a composition containing active cells according to the invention and in a second compartment an additional excipient and / or active, the compositions contained in said first and second compartments being here considered as a combination composition for simultaneous, separate use or spread over time in particular in one of the treatments defined above. The treatment method according to the invention is more particularly adapted to a restorative cosmetic repair and smoothing treatment on an aged skin, in particular at the level of the falling surface of the neck 10 and the fold of the valley of tears. The present invention will be better understood and other advantages will become apparent in light of the following detailed description of an example of plant cell preparation, and in vitro and in vivo tests performed on these cells. A) Example of plant cell preparation Creation of a cell line Selected pieces of leaves of the genus Leontopodium alpinum are removed, washed and cut into small pieces of a few mm, so as to make 200 to 1500 explants. After a series of decontamination and sterilization treatments, the pieces are placed on an agar culture medium in the presence of a nutrient medium containing plant growth hormones in order to induce callogenesis (callus formation). After a suitable period of time, a cluster of dedifferentiated or cal cells is formed, which is transferred over a larger area and into a fresh culture medium in order to multiply. A number of subcultures (transfers on a fresh culture medium) are carried out to stabilize the cell line, i.e. until it exhibits a high and constant proliferation rate, conservation. phenotype, a constant content of bioactive compounds of interest (primary and secondary metabolites). The cell line is then subjected to a selection step which consists in culturing the cells for a suitable duration, taking the aggregates of cells formed and inoculating them in a liquid culture medium for a duration allowing to obtain the multiplication of the cells. cell aggregate.
    [0020] The best cell line will be that to obtain as rapidly as possible and reproducibly a large biomass having an optimal content of selected metabolites, the best biological activity and a homogeneous phenotype. This was also chosen for its ability to produce lactic acid A and B in an amount of about 5% by weight, based on the dry weight of the cells, as measured by HPLC.
    [0021] INDUSTRIAL PROCESS FOR THE PRODUCTION OF AN INDIFFERENTIATED OR DEDIFFERENCED CAPITAL OF LEONTOPODO ALIMINUM AND THE TREATMENT OF THIS BIOMASS Starting from a cell line prepared as described above or from an existing line.
    [0022] The Leontopodium alpinum line is initially multiplied to obtain a sufficient amount of dedifferentiated cell biomass in order to perform the large scale production step. The following steps are carried out: a) Inoculation of the selected line in a liquid medium and culture for a time sufficient to obtain a biomass increase of at least 300%; b) Optionally, transfer of the suspension obtained in a) into a fresh liquid medium and again culture sufficient time to obtain a biomass increase of at least 300%; c) optionally, repeating step b); D) Transfer the cell suspensions obtained in steps a) to c) into a bioreactor with fresh liquid medium, and conduct the culture under such conditions and for a time sufficient to obtain a cellular biomass containing the metabolites of interest c that is, phenylpropanoides glycosides and leontopodic acids in sufficient amounts, this bioreactor production step comprising an elicitation step performed by modifying the nutrient levels of the culture medium. Bioreactor: Volume: 5 to 50 times larger than the volume of biomass used as inoculum; internal surface of the bioreactor smooth and uniform (no edges or angles can cause rupture of cell walls).
    [0023] Culture conditions: Culture medium: medium comprising inorganic salts (solution of macroelements and microelements), vitamins, plant hormones and sucrose. Vegetable agar is added in the solid media. Temperature: between 15 ° C and 35 ° C, preferably between 20 ° C and 30 ° C and even more preferably at 25 ° C. Duration: between 7 and 21 days, preferably between 10 and 14 days. Stirring of biomass: it is important that the biomass is aerated optimally, and that at the same time, it is kept agitated either by an internal means, or by an external means. It is necessary that the agitation, although low, be effective, especially in the final stages, when the biomass is in large quantity. For the purposes of the present invention, suitable internal stirring means are propellers rotating between 20 and 120 rpm, preferably at 60 rpm, or externally orbital stirring means preferably rotating. between 40 and 200 rpm and preferably about 120 rpm. Oxygenation: normally performed using sterile air, at a rate of 0.5 to 4 liters per minute, preferably between 2 and 2.5 liters per minute, for a volume of 10 liters of biomass. Alternatively, gas mixtures containing from 10% to 100% v / v of oxygen may be used. It will be preferable to use means for diffusing air or oxygen with a nozzle having a flow rate of between 10 ml / min and 600 ml / min and preferably between 50 ml / min and 350 ml. / min. Treatment of the biomass obtained Filtration to eliminate the culture medium and recover the biomass of cells. This biomass can be characterized by its equivalent level of lyophilized cells. Homogenization under high pressure of the cellular biomass: allows a reduction of the size of the cellular aggregates; some cells may be exploded and can then be in the presence of a mixture of whole cells and ground cells, preferably keeping 10% of whole cells. Characterization of the active compounds contained in the cells by analytical determination of 10 primary and secondary metabolites produced by the culture, including the protein content, phenylpropanoid glycosides, of which lenopodic acids A and B. Optionally: 1) drying of the cells, in particular by lyophilization or atomization to enable greater stability of the compounds of interest, improve long-term storage without having to add preservatives. 2) extraction of the cell contents by grinding / lysis / bursting of the cells and separation of the liquid and solid phases (by centrifugation or filtration or other), in order to obtain a particular cell extract. 3) Purification of the cell extract to increase the content of leontopodic acids.
    [0024] B) Preparation of an active ingredient for use according to the invention The biomass of cells, either as obtained above after filtration, or in dried form, or the resulting extract, can be mixed with one another. physiologically acceptable medium forming the excipient. As a preferred example, this physiologically acceptable medium is a hydrophilic matrix in which the cells are suspended, for example in the case of a cosmetic composition of glycerol and / or butylene glycol. Additives may also be added if necessary, such as antimicrobial agents, antioxidants, stabilizing agents, agents acting on the pH, emulsifying agents or thickeners, especially a thickener such as xanthan gum which will promote the keeping the cells in suspension.
    [0025] An active ingredient for cosmetic use can thus be formed for the implementation of the invention, comprising, for example, 20% by weight of fresh biomass of whole dedifferentiated cells (corresponding to about 1% of dry cells), in an excipient mixture. physiologically acceptable composition consisting of glycerol (about 80%) and xanthan gum (0.3% by weight), said ingredient having a final level of about 0.05% of luteopodic acids A and B (about 15%). A acid and 85% B). This ingredient is then usable to prepare cosmetic formulations as set forth below in Galenic point F).
    [0026] It goes without saying that, according to the invention, it is possible to use plant cells comprising a different level of leontopodic acids, in particular higher, or obtained directly by means of the in vitro method (for example by means of an appropriate elicitation which makes it possible to increase the rate), or obtained by a purification / concentration phase of the cells obtained (for example by a concentration step after extraction of the cell contents). By way of example, it is possible to manufacture plant cells in the form of a purified extract comprising a high level of leotopodic acids, for example a level of 25% relative to the dry matter, said cells being themselves used to make an active ingredient as explained above.
    [0027] C) In vitro test results The in vitro tests were carried out using an Ethanol / Water extract (70/30) of lyophilized cells from the Leontopodium Alpinium cell culture (21% of lyophilized cells obtained according to the example). above in the solvent mixture). This solution is the mother solution. If 0.2% of this solution is taken in a test medium, the resulting solution will be 420ppm cell equivalent. Hereinafter, we will speak of a cell extract with X equivalent cells. 1) Restoration of dermal cell homeostasis The tests below were carried out using, if necessary, experimental models of accelerated aging (by UVA or UVB irradiation) in order to demonstrate the interest of the product according to the invention for the restoration of homeostasis of dermal cells. A) Synthesis of thrombospondins (TSP) Principle: Normal human keratinocytes (KHN) in culture were brought into contact with the cell extract according to the invention at 140 and 280 ppm of cell-equivalent for 48 h. The syntheses of TSP-1 and -2 were evaluated by an ELISA method in the cell culture medium. Table 1: Production of TSP-2 by KHN +/- the cell extract according to the invention (n = 3). TSP-2 (ng / 106 cell.) Variation (%) Control 5.87 ± 0.28 Reference 140ppm 7.57 ± 1.37 + 29%; dns 280ppm 9.2 ± 0.32 + 57%; These results show that the cell extract according to the invention has 280 ppm of cell equivalent significantly induces the production of TSP-2 in the KHN (+ 57%, p <0.01). This increase in TSP makes it possible to limit the activity of MMPs with the consequence of improving the quality of the dermis. b) Reduction of the synthesis of stress-induced MMPs Principle: Human dermal fibroblasts (FHD) in culture were brought into contact with the cell extract according to the invention (at 70; 140 or 280 ppm cell equivalent) for 24 hours then, after rinsing, the carpets were exposed to UVA irradiation in order to increase the pool of MMP production by these cells and thus to model accelerated aging of the cells. The cells were then put back into the culture medium in contact with the extract according to the invention for 24 hours. The production of MMPs in the culture medium was monitored using Multiplex / ELISA methods. Table 2: Variation of MMP production following UVA irradiation on fibroblasts in contact with the cell extract according to the invention (n = 4). MMP-1 Variation MMP-7 Variation MMP-9 Variation (pg / mL / 106cc) (%) (pg / mL / 106cc) (%) (pg / mL / 10 6 (%) cel) Control 1622 ± 47 Reference 657 ± 196 Reference 22.5 ± 3.1 Reference 7Oppm ± -42%; 388 ± 136 -41%; 18.8 ± 2.2-17%; 942 82 p <0.01 p <0.07 dns 140ppm 794 ± 55 -51%; 266 ± 134 p-5 <90%, 02; 16.8 ± 1.9-25%; p <0.01 p = 0.02 280ppm 762 ± 40 -53%; 289 ± 159-56%; 17.8 ± 18-21%; p <0.01 p <0.03 'p <0.04 UVA stress, modeling aging, applied to fibroblasts, significantly increased the production of MMP-1 (+ 292%, p <0.01), MMP-7 (+ 48%, p = 0.08) and MMP-9 (+ 51%, p <0.01). The contact of the cells with the extract according to the invention with 280 ppm of equivalent-cells made it possible to strongly reduce the induction of MMP-1 (-53%, p <0.01), MMP-7 (-59% , p <0.02) and MMP-9 (-25%, p = 0.02). Thus, the extract according to the invention makes it possible to limit the formation of proteases that fragment the extracellular matrix of the dermis. c) Increased synthesis of induced stress TIMPs Principle: In parallel with the previous trial, the production of TIMP was monitored using Multiplex / ELISA methods in the culture medium.
    [0028] Table 3: Variation of the TIMP production following UVA irradiation on fibroblasts in contact with the cell extract according to the invention (n = 4). TIMP-1 Variation (%) TIMP-2 Variation (%) (ng / mL / 106cell.) (Ng / mL / 106 cell) Control 34.5 ± 9.0 Reference 10.3 ± 2.4 Reference 140ppm 87 , 6 ± 17.6 + 154%; p <0.01 22.1 ± 2.9 + 115%; p <0.01 280ppm 93.0 ± 2.5 + 170%; p <0.01 24.8 ± 0.9 + 141%; p <0.01 TIMP-3 Variation (TIMP-4 Variation (ng / mL / 106 cell) (%) (pg / mL / 106 cell) (%) Control 16.2 ± 3.2 Reference 294 ± 45 Reference 140ppm 26.1 ± 5.3 + 61%, p <0.02 388 ± 35 + 32%, p <0.02 280ppm 28.4 ± 3.7 + 75%, p <0.01 402 ± 11 + 37%; 0.01 The contact of the cells with the cell extract according to the invention at 280 ppm of cell equivalent made it possible to reactivate the induction of TIMP-1 (+ 170%, p <0.01), TIMP-2 ( + 141%, p <0.01), TIMP-3 3031454 (+ 75%, p <0.01), and TIMP-4 (+ 37%, p <0.01). MMP-specific inhibitors are strongly induced by the invention d) Effect according to the invention of the extract on the basal production of dermal macromolecules 5 - Collagen I Human dermal fibroblasts (FHD) are inoculated. Once the confluence is reached, the cells are brought into contact with the VitC +/- extract according to the invention at 140 ppm equivalent cells and put back into culture for 72 hours. The supernatants are removed and the cell mats extracted to quantify the number of cells. The collagen I assay is performed on the supernatants by ELISA method. No toxicity was noted with respect to the control. Table 4: Synthesis of Collagen I on FHD (n = 5) [Co111] ng / 10E6 cells Variation (%) Control 11994.4 ± 347.8 Reference 140ppm 16385.8 ± 487.1 36.6; p <0.01 - Collagen IV Human dermal fibroblasts (HDF) are inoculated. Once the confluence is reached, the cells are brought into contact with the VitC +/- extract according to the invention at 140 ppm equivalent cells and put back in culture for 72 hours. The supernatants are removed and the cell mats extracted to quantify the number of cells. The collagen IV assay is performed on the supernatants by ELISA method. No toxicity was noted with respect to the control. Table 5: Synthesis of Collagen IV on FHD (n = 5) [Co114] ng / 10E6 cells Variation (%) Control 7.8 ± 0.5 Reference 140ppm 12.6 ± 1.7 61.5; p <0.01 - Hyaluronic acid Dermal human fibroblasts (FHD) are inoculated. Once the confluence is reached, the cells are brought into contact with the cell extract according to the invention at 210 ppm and 420 ppm of equivalent cells, then put back into culture for 72 hours. The supernatants are removed and the cell mats extracted to quantify the number of cells. The determination of hyaluronic acid is carried out on the supernatants by ELISA method. No toxicity was noted with respect to the control.
    [0029] Table 6: Synthesis of hyaluronic acid (HA) by FHD (n = 5) [HA] ng / 10E6 cell. Variation (%) Control 2546 ± 373 Reference 210ppm 3068 ± 190 20.5; p <0.05 420ppm 4067 ± 1123 59.7; p <0.05 3031454 19 For the 3 macromolecules tested, there is a stimulation of their production by fibroblasts in the presence of the cell extract according to the invention. 2) Improvement of cellular and energetic dynamism a) Contraction of modeled dermas Principle: HDF in culture were accelerated with H202. The cells were then put back into their culture medium for 24 hours. Then, these cells were included in a dermis model and, after polymerization, received the cell extract according to the invention (140 or 280ppm of equivalent cells). The contraction of the dermis model was followed for 4 days, photographs allowed to quantify the differences by image analysis.
    [0030] Table 7: Modulation of contraction of a dermal model by FHD +/- cell extract according to the invention (n = 6). Contraction Variation (%) Variation (%) (arbitrary units) Without H202 5143 ± 1157 Reference H202 3459 ± 488 -33%; p <0.01 Reference H202 + 140ppm cell equivalent 4114 ± 346 - + 19%; p <0.05 H202 + 280ppm cell equivalent 5319 ± 925 - + 54%; p <0.01 These results confirm that the contact of the cells with H2O2 makes it possible to reduce the contractile capacities of these cells (-33%, p <0.01). The extract according to the invention at 280 ppm of cell equivalent makes it possible to reduce, in a dose-dependent manner, the effect of accelerated aging on the contractile capacity of the cells (+ 54% contraction compared to control; , 01). b) Mitochondrial dynamics Principle: Older cells are more retracted than young cells. The preservation of the shape of the cell as well as the appearance of its mitochondrial network are therefore good indicators of their dynamism. Human dermal fibroblasts (HDF) in culture were stressed using UVB exposure to obtain a retracted phenotype and a fragmented mitochondrial network. Just after irradiation, the cells were then returned to their culture medium with the cell extract according to the invention (280 ppm of equivalent cells) and fixed after 18 hours. The cells were then labeled with the Mitotraker specific dye and the network was quantified from photos by first extracting the network connections using a specific computer tool. The photos show that exposure, even minor HDF UVB, causes their retraction and fragmentation of their mitochondrial network (less mitochondria filaments, more punctate structures). The contact with the cell extract according to the invention, placed just after the irradiation, makes it possible to normalize the appearance of the cells by maintaining their non-retracted phenotype and to give the mitochondrial network a less fragmented aspect.
    [0031] Table 8: Modulation of mitochondrial connections by FHD +/- cell extract according to the invention (n = 28). Connections Variation (%) Variation (%) mitochondrial (arbitrary units) Not irradiated 133 ± 45 UVB reference 104 ± 41 -22%; p <0.02 UVB reference + 280ppm cell equivalent 136 ± 70 + 31%; p <0.05 These results show that the mitochondrial network is strongly disturbed several hours after UVB irradiation, even weak (22% fewer connections, p <0.02). The contact with the cell extract according to the invention placed after the irradiation makes it possible to maintain the network and the connections at the non-irradiated control level (+ 31%, p <0.05 vs UVB alone). The cell extract according to the invention therefore makes it possible to restore cellular and mitochondrial dynamism. D) Galenic Various cosmetic formulations are described below. Additional active ingredients, optionally in support of and / or in addition to the activity of the active ingredient according to the invention based on undifferentiated or dedifferentiated Leontopodium alpinum cells, may be added in the appropriate phase according to their hydrophobic or hydrophilic nature. These ingredients can be of any category depending on their function (s), the place of application (body, face, neck, bust, hands, etc.), the desired end effect and the targeted consumer, for example specific anti-wrinkle, moisturizing, concealer, firming, anti-glycation, volumizing, soothing, myo-relaxing, anti-redness, detoxifying, etc. Active ingredient used in the galenical formulations given below: 20% by weight, based on the total weight of the composition of the ingredient, of undifferentiated or dedifferentiated fresh cells (biomass) according to the invention (corresponding to approximately 1% of dry cells), the ingredient having a final content of about 0.05% liposome A and B in a physiologically acceptable excipient mixture of glycerol, xanthan gum thickener and citric acid to adjust the pH of the optionally. This ingredient is recommended between 0.1 and 10%, preferably between 1 and 5%, more preferably between 2 and 3% and usually 2%. 1) "Cream" form, well suited in particular for the neck PRODUCT% INCI NAME Phase A Qsp100 Water Carbomer 0.30 H20 Optasense G83TM 25 3031454 21 Phase B Brij S2-SS- (RB) TM 0.40 Steareth-2 Brij S10 -S0- (RB) TM 1.20 Steareth-10 Crodafos CES-PA- (RB) TM 4.00 Cetearyl Alcohol & Dicetyl Phosphate & Ceteth-Phosphate Crodacol CS 90-PA- (RB) TM 1.50 Cetearyl Alcohol Laurocapram 2.50 Laurocapram BRB CM 56TM 2.00 Cyclopentasiloxane & Cyclohexasiloxane Crodamol OSU-LQ- (RB) TM 7.00 Diethylhexyl Succinate Phase C Glycerin 4.00 Glycerin Octanediol 0.50 Caprylyl Glycol Phase D Phenoxyethanol qs Phenoxyethanol Phase E Sorbate potassium qs Potassium Sorbate Phase F H20 4.00 Water NaOH 30% 0.40 Sodium Hydroxide Phase G Active ingredient according to the invention 1.00 to 5.00 / Phase H Fragrance 0.10 Fragrance Protocol: weigh phase A and leave swell without stirring for 30 min. Put phase A to heat at 75 ° C in a water bath. Weigh phase B and heat to 75 ° C in a water bath. Mix well. Weigh and melt phase C at 45 ° C. Add phase D to phase C, previously cooled. Pour phase C + D in phase A, with stirring staro v = 500 tlmn. Well homogenize.
    [0032] Pour phase B into the previous phase, with stirring staro v = 1000 rpm. Well homogenize. Extemporaneously, add phase E, well homogenize. Add phase F, mix well. Add phase G, below 45 ° C, mix well, 1 hour. Add phase H, mix well. Examples of ingredients that can be added to this formulation: 10-MATRIXYL synthe'6TM: peptide-based anti-wrinkle active ingredient marketed by Sederma (WO2010 / 082175), which helps repair skin damage caused by aging. - MATRIXYL 3O00TM: peptide-based anti-wrinkle active ingredient marketed by Sederma (WO2005 / 048968), which helps repair the skin damage caused by aging. - PRODIZIATM: active ingredient marketed by Sederma, which combats the cutaneous signs of fatigue caused by glycation and glycoxidation. 2) Form "Serum" PRODUCT% INCI NAME Phase A Qsp100 Water qs Potassium Sorbate H20 Potassium Sorbate 3031454 22 Phase B Butylene Glycol 3.00 Butylene Glycol Phenoxyethanol qs Phenoxyethanol Keltrol CG-SFTTM 0.30 Xanthan Gum Satiagel VPC 614TM 0.20 Chondrus Crispus (Carrageenan) Extract Fragrance 0.10 Fragrance Phase C H20 0.20 Water Lactic Acid 0.02 Lactic Acid Phase D Active Ingredient according to the invention 1.00 to 5.00 / Protocol: Weigh phase A and put under propeller stirring v = 300 rpm. Weigh and homogenise phase B. Add phase B to phase A with rapid stirring v = 600 rpm. Leave to homogenize for 1 hour. Adjust the pH to 6.00 ± 0.10 with phase C. Add phase D in the previous phase and mix thoroughly.
    [0033] Examples of ingredients that can be added to this formulation: - KOMBUCHKATm: active ingredient marketed by Sederma (WO2004 / 012650), having an effect on the radiance of the complexion, anti-glycation, redensifying the adipocyte population, and improving the quality skin (smoothing, shine). - REVIDRATTm: active marketed by Sederma, which in particular improves the cohesion of the epidermis and its hydration. - IDEALIFTTm: active ingredient marketed by Sederma (W02010 / 136965) that fights flaccidity of the face and improves resistance to gravity. 3) "Lotion" form, well adapted especially for the neck PRODUCT% INCI NAME Phase A H20 Qsp100 Water Potassium Sorbate 0.10 Potassium Sorbate Phase B Glycerine 2.00 Glycerin Phenoxyethanol qs Phenoxyethanol Keltrol CG-SFTTM 0.60 Xanthan Gum Vivapur CS 032TM 0.25 Microcrystalline Cellulose (and) Xanthan Gum Phase C Span 40-PW- (MV) TM 1.00 Sorbitan Palmitate Span 60-PW- (MV) TM 1.50 Sorbitan Stearate Crodamol GTCC-LQ- (MV) TM 4.00 Caprylic / Capric Triglyceride Prisorine 3505TM 4.00 Isostearic Acid Phase D H20 0.30 Water Lactic Acid 0.03 Lactic Acid Phase E Active Ingredient according to the invention 1.00 to 5.00 / Phase F Perfume 0, 10 Fragrance 3031454 23 Protocol: Weigh phase A. Weigh and mix phase B. Add phase B to phase A, with propeller stirring v = 800 rpm. Well homogenize. Put phase A + B to heat at 75 ° C in a water bath. Weigh and put phase C to heat to 75 ° C in a water bath. Pour phase C into phase A + B, with very strong staro stirring v = 1000 rpm; well homogenize. Adjust the pH to 5.60 ± 0.10 with phase D 5 below 35 ° C. Add phase E; well homogenize. Add phase F; well homogenize. Examples of ingredients that can be added to this formulation: - PACIFEELTM: Soothing active ingredient marketed by Sederma including an extract of the plant Mirabilis Jalapa, which soothes and improves the comfort of sensitive and reactive skin. - RESISTEMTm: anti-aging ingredient marketed by Sederma (WO2012 / 104774), helping skin 10 to build its own anti-aging defense system, based on an extract obtained by cell culture of the plant Globularia cordifolia. - MEIRITAGETm: anti-aging active ingredient based on three plant extracts (Astragalus membranaceus (Huang Qi), Bupleurum falcatum (Chai Hu) and Atractylodes macrocephala (Bai Zhu), which improves the uniformity and radiance of the complexion. 4) Form "Gel" for the eye contour, especially suitable for the "valley of tears" PRODUCT% INCI NAME Phase A H20 Qsp100 Water Potassium Sorbate 0.10 Potassium Sorbate Sodium Disulfite 0.01 Sodium Sulfite Phase B Keltrol GC-SFTTM 0.50 Xanthan Gum N-HANCE HP4OTM 0.25 Hydroxypropyl Guar ZemeaTM 1.90 Propanediol Phenoxyethanol qs Phenoxyethanol Phase C Crodamol ISIS-LQ- (MV) TM 2.90 Isostearyl Isostearate Span 20-LQ- (SG) TM 0.70 Sorbitan Laurate Ethanol 4.80 Ethanol Fragrance 0.10 Fragrance Phase D H20 2.00 Water Lactic Acid 0.02 Lactic Acid Phase E Active Ingredient according to the invention 1.00 to 5.00 / Protocol: Weigh the phase A and homogenize well. Weigh phase B and mix well. Add phase B to phase A slowly with propeller stirring v = 600rpm, for 30 min. Weigh phase C and mix. Add phase C to phase A + B with vigorous stirring Staro v = 3000rpm. Adjust the pH to 5.50 ± 0.10 with phase D. Add phase E in the previous phase and mix thoroughly.
    [0034] Examples of ingredients that can be added to this formulation: MATRIXYL synthe'6TM: peptide-based anti-wrinkle active ingredient marketed by Sederma (W02010 / 082175) which helps repair the skin damage caused by aging. - HALOXYLTM: active commercialized by Sederma (W02005 / 102266), which improves the contour of the 5 eyes by resorbing the rings. 3% of this ingredient can for example be added at the end of the formulation. - EYELISSTm: is an active commercialized by Sederma (W02003 / 068141), which helps to prevent and fight the appearance of the bags under the eyes. 3% of this ingredient can for example be added at the end of the formulation. 5) Form serum "foundation" with sunscreen PRODUCT% NAME INCI Phase A H20 Qsp100 Water Potassium sorbate 0.10 Potassium Sorbate Phase B Glycerin 2.00 Glycerin Phenoxyethanol qs Phenoxyethanol Keltrol CG-SFTTM 0.60 Xanthan Gum Vivapur CS 032TM 0.25 Microcrystalline Cellulose & Xanthan Gum Phase C Eusolex 4360TM 1.80 Benzophenone-3 Eusolex 2292TM 3.50 Ethylhexyl Methoxycinnamate & BHT Span 40-PW- (MV) TM 1.00 Sorbitan Palmitate Span 60-PW- (MV ) TM 1.50 Sorbitan Stearate Crodamol GTCC-LQ- (RB) TM 1.35 Caprylic / Capric Triglyceride Prisorine 3505TM 1.35 Isostearic Acid Phase D H20 0.20 Water Lactic Acid 0.02 Lactic Acid Phase E Active Ingredient invention 1.00 to 5.00 Phase H Fragrance 0.10 Fragrance Protocol: Weigh and apply phase A with propeller stirring v = 500rpm. Weigh phase B and mix well. Add phase B to phase A with v-propeller stirring; well homogenize. Put phase A + B to be heated to 75 ° C in a water bath. Weigh phase C and heat to a water bath at 75 ° C. Add phase C to phase A + B with staro stirring v = 1000 rpm. Homogenize 30 min. Adjust the pH to 5.80 ± 0.1 with phase D, below 35 ° C. Add phase E, mix well. Add phase F, homogenize well. Eusolex 4360TM is a UVA filter and Eusolex 2292TM a UVB filter. Examples of ingredients that can be added to this formulation: DERMAXYL ™: anti-aging active ingredient marketed by Sederma (W02004 / 101609) which smoothes wrinkles and repairs the cutaneous barrier. - VENUCEANETM: active ingredient marketed by Sederma (WO2002 / 066668) which prevents visible signs of photoaging (spots, wrinkles, dryness, etc.), protects cell structures from damage caused by UV radiation and reinforces the integrity of the skin. skin. - MELASLOW ™: active marketed by Sederma which promotes the lightening of the complexion and the depigmentation of the spots (Mandarin extract of Japan Citrus reticulata Blanco var.nshiu). 6) Form acidic pH cream PRODUCT% INCI NAME Phase A H20 Qsp100 Water Potassium Sorbate 0.10 Potassium Sorbate Phase B Glycerin 5.00 Glycerin Phenoxyethanol qs Phenoxyethanol Keltrol CG-SFTTM 0.60 Xanthan Gum Supercol GF 0.25 Cyamopsis Tetragonoloba ( Guar) Gum Phase C Span 40-PW- (MV) TM 1.00 Sorbitan Stearate Span 60-PW- (MV) TM 1.50 Sorbitan Palmitate Crodamol IPIS-LQ- (MV) Tm 4.00 Isopropyl Isostearate Prisorine 3505TM 4 , 00 Isostearic Acid D Phase H20 Qsp100 Water Lactic Acid 0.04 Lactic Acid Phase E Active Ingredient according to the invention 1.00 to 5.00 Phase F Fragrance 0.10 Fragrance Phase G FruitbioTM 0.90 Water (Aqua) - Lactic Acid - Camellia sinensis leaf extract - Glycerin - Citric Acid - Malic Acid Protocol: Weigh phase A and put under stirring propeller v = 130rpm. Weigh and homogenize phase B. Add phase B to phase A with stirring v = 400 rpm, mix well for 30 min. Heat the phase A + B to 75 ° C in a water bath. Weigh phase C and heat to 75 ° C in a water bath. Add phase C in phase A + B, with stirring staro v = 1000rpm; homogenize 30 min. Adjust the pH to 5.60 ± 0.10 with phase D, below 35 ° C. Add phase E in the previous phase, mix well. Add phase F, homogenize well. Adjust the pH to 3.50 ± 0.10 with phase G. FruitbioTM is an active ingredient marketed by Sederma consisting of a complex of ahydroxy acids associated with a green tea extract.
    [0035] Examples of ingredients that can be added to this formulation: CALMOSENSINETm: sedative active ingredient marketed by Sederma (W01998 / 07744) containing Tyr-Arg lipo-dipeptide. It reduces feelings of discomfort. - PHYTOTONINETM: Active ingredient marketed by Sederma comprising a synergistic combination of three plant active ingredients, Arnica montana flower flavonoids, 3031454 Polygonatum multiflorum rhyzome saponosides (Solomon seal) and Cupressus sempervirens cone proanthocyanidins (Cypress) ; significantly improves the appearance of "blotchy" skin. CHROMOCARETM: an anti-aging active combining an extract of rabdosia rubescens rich in oridonine and a Siegesbeckia orientalis extract rich in darutoside, marketed by Sederma 5 (WO2010 / 119423) which unifies and rejuvenates the complexion. 7) Thick cream form, especially for a night cream PRODUCT% INCI NAME Phase A H20 Qsp100 Water Magnesium Sulfate 1.10 Magnesium Sulfate Phase B Glycerine 5.00 Glycerin Phenoxyethanol qs Phenoxyethanol Phase C Beeswax BP-EP 139901TM 2.00 Cera Alba Crodamol MM-PA- (RB) TM 3.00 Myristyl Myristate Arlacel 986-S0- (MV) TM 3.50 Sorbitan Isostearate & Hydrogenated Castor Oil & Cera Alba & Stearic Acid Cithrol PG32IS-LQ- (MV) TM 1, 50 Polyglyceryl-3 Diisostearate Crodamol GTCC-LQ- (MV) TM 7.00 Caprylic / Capric Triglyceride Crodamol GTIS-LQ- (MV) TM 2.00 Triisostearin Squalane 6.00 Phase D Ceramide 2TM Ceramide NG Crodamol ISIS-LQ- ( MV) TM Isostearyl Isostearate Phase E Active Ingredient according to the invention 1.00 to 5.00 / Phase F Fragrance 0.10 Fragrance Protocol: Weigh phase A. Weigh and mix phase B. Add phase B to phase A and mix well. Put phase A + B to heat at 75 ° C in a water bath. Weigh phase C and put it to heat at 75 ° C in a water bath. Well homogenize. Heat phase D at 90 ° C on a heating plate while stirring with a spatula. Add phase D to phase C at 75 ° C in a water bath. Add phase E to phase A + B at 75 ° C in a water bath. Slowly add phase A + B + E to phase C + D with strong stirring (s = 2500 rpm, then 1500 rpm without water bath). Add phase F. CERAMIDE 2TM is an active ingredient marketed by Sederma, consisting of a pure substance identical to the ceramides present in the skin, which constitute 40 to 50% of the cutaneous lipids.
    [0036] This ingredient has a restructuring function, maintains the hydration and integrity of the skin barrier. E) In Vivo Tests The evaluation of the in vivo efficacy of the treatment according to the invention was carried out on the neck and on the fold of the valley of tears.
    [0037] Principle The study was carried out on women of 59 years of age (41 - 71 years old) with several visible signs of age: - on the neck with a measurement of the falling surface, by image analysis on standardized photographs, as well as an expert evaluation on these same photographs. - on the fold of the valley of tears ("tear trough" in English, hollow in which the tears flow before rolling on the cheeks), with a measurement of its surface by projection of fringes. For the neck, a panel of 31 middle-aged 60-year-olds (aged 45-71) was selected on the basis of the visible importance of the falling surface.
    [0038] 10 For the valley of tears, 21 people, over 55 years old, were included on the basis of the visible importance of this type of fold (average age 64 years (56 - 71 years)). Protocol Specific inclusion criteria Volunteers had to have a certain falling surface of their neck skin and present a clearly visible fold in the area known as the Valley of Tears. Hormonal consistency during the 3 months preceding the test and during the test was requested. A wash-out period of 7 days was required, during which the volunteer used only a moisturizer on the face. During the test, moderate solar exposure and the exclusive use of the supplied products was requested. Type of study and duration The study was conducted in single blind on the neck and face. Each volunteer applied a cream according to the invention (formula of Example 1 of the galenic at 2%) and a contra-lateral placebo cream on his face, except for the neck where, for practical reasons, they do not applied that 2% cream according to the invention. The 2 creams were applied in bi-daily massage for 6 weeks. The synopsis of the study can be summarized according to the diagram below.
    [0039] 25 TO T 3 weeks T 6 weeks Dermatop, fold of the Dermatop Valley, fold of the valley tears tears Photos face & neck Photos neck Photos face & neck Statistical studies were performed using Student's t test or if necessary, with a non-parametric Wilcoxon test. Unilateral tests were performed on matched series. Study of the falling surface of the neck and smoothing The effectiveness of the product according to the invention on the neck consisted in measuring the falling part of the neck by an analysis of standardized photographic images, and in an expert evaluation on these same photographs.
    [0040] 35 Photos were taken at TO and T3weekly using a photographic bench (Société Orion Concept, France) equipped with a high-definition digital camera, specific lighting and a system for restraining volunteers. The position of the face, the photo and lighting parameters were standardized and controlled in order to be reproduced over time.The falling surface of the neck was measured on a photo taken in profile, by image analysis. immovable (scars, spots, etc.) were used to precisely define the contours of the neck surface including the falling surface.
    [0041] 3031454 28 Table 9: Area (in mm2) TO T3sem. Mean ± E-type 101.2 ± 40 90.5 ± 42% variation vs.TO reference -10.6% Significance vs. TO Responders p <0.01 77% These results show a decrease of 3 weeks from the falling surface of the neck; this decrease reached about 11% (p <0.01) of the area measured at TO. In parallel, a high responder rate was observed (77%). Regarding the smoothing effect of the skin of the neck, it was appreciated by a panel of 7 expert judges 5 who visualized the photos of the 28 volunteers with folds on the neck. Experts found a smoothing effect in 56% of cases after 6 weeks. Study of the fold of the valley of tears The valley of tears ("tear trough" in English) is the hollow in which the tears flow before rolling on the cheeks. This sub-orbital hollow is continuous with the lower eyelid and will sometimes extend between the cheek and the cheekbone. It widens significantly with the age from which the increase of its volume. The factors generally implicated are fatty melt and flaccidity. In order to test the effectiveness of the treatment according to the invention on this site, a FOITS contactless system ("Fast Optical In vivo Topometry System") was used. This system is based on the analysis of the projection of optical fringes on the cutaneous zone studied, here the valley of tears. The apparatus used (Société Dermatop, Eotech, France) consists of a projector and a camera that are integral and form a precise angle, allowing triangulation. The study of the deformation of the fringes by the relief of the zone allows a 3D reconstruction of the relief. The objectification of the effect according to the invention was made, using Aeva ™ software (Eotech, France), by analyzing the data on the extent of the valley of tears measured by its circumference. In parallel, the volume parameter of this valley was obtained by the same technique. Table 10: Variation in the extent of the valley of tears and volume after application according to the invention. (N = 21 volunteers,> 55 years) Extent of the Valley of tears Volume of the valley of tears (mm) (mm3) According to the invention Placebo According to the invention Placebo TO T6 TO T6 TO T6 TO T6 Mean ± E-type 55.6 50.6 51.6 51.6 6.8 6.2 7.4 7.8 ± 15.1 ± 14.0 ± 13.0 ± 14.1 ± 5.2 ± 4.9 ± 6 , 9 ± 6.7 variation vs. TO x -9% x 0.0% X -8.8% x + 5.4% 3031454 29 Significance vs. TO p <0.01 dns p = 0.082 dns -33% X -84% X Maximum 86% x 62% x Responders Significance vs. Placebo x p <0.01 x x X p <0.01 x x dns: not significant difference. The results show that the treatment according to the invention leads to a 9% reduction in the extent of the valley of tears (measured in circumference), this reduction being significant compared to placebo (p <0.01). In contralateral application of a placebo produces no significant change. In addition, the volume of the valley of tears of those over 55 years old, is reduced by 8.8%, this reduction being significant (p <0.01) compared to placebo.
    权利要求:
    Claims (14)
    [0001]
    REVENDICATIONS1. Use of undifferentiated or dedifferentiated plant cells of Leontopodium alpinium obtained by an in vitro cell culture method for a non-therapeutic cosmetic treatment to restore the homeostasis of aged skin cells and increase their metabolic and energetic activity.
    [0002]
    2. Use according to claim 1 of whole plant cells and / or lysed.
    [0003]
    3. Use according to claim 2 of a cell extract of said whole cells and / or lysed.
    [0004]
    4. Use according to one of claims 1 to 3, plant cells in dried form.
    [0005]
    5. Use according to claim any one of claims 1 to 4, a cosmetic composition comprising said plant cells suspended and / or solubilized in a physiologically acceptable medium.
    [0006]
    6. Use according to claim 5, characterized in that the physiologically acceptable medium is a hydrophilic matrix.
    [0007]
    7. Use according to one of claims 1 to 6, characterized in that the treatment is topical.
    [0008]
    8. Use according to one of claims 1 to 7, for a tensor repair treatment and smoothing on aged skin.
    [0009]
    9. Use according to claim 8, for a cosmetic treatment of the falling surface of the neck and / or the fold of the valley of tears.
    [0010]
    10. Use of undifferentiated or dedifferentiated plant cells of Leontopodium alpinium obtained by an in vitro cell culture method for the manufacture of a non-therapeutic cosmetic active ingredient for restoring homeostasis of aged skin cells and increasing their metabolic and energetic activity .
    [0011]
    11. Use according to claim 10, characterized in that the active ingredient comprises at least 0.04% of léontopodiques acids A and B.
    [0012]
    12. Cosmetic active ingredient, for use according to one of claims 1 to 9, comprising undifferentiated or dedifferentiated plant cells of Leontopodium alpinium obtained by an in vitro cell culture method and comprising at least 0.04% of leprosy acids. A and B in a physiologically acceptable matrix.
    [0013]
    13. Ingredient according to claim 12, characterized in that the plant cells are whole and / or lysed.
    [0014]
    14. Ingredient according to claim 13, characterized in that it comprises a cell extract of said whole and / or lysed cells.
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    EP2319914A1|2009-11-09|2011-05-11|I.R.B. Istituto Di Ricerche Biotecnologiche S.r.l.|Preparation and use of plant meristem cells with a high content of caffeic acid derivatives|EP3607997A1|2018-08-10|2020-02-12|Société De Recherche Cosmétique S.à.r.L.|Cosmetic composition comprising an extract of passion flower and edelweiss cells and uses|
    WO2021064173A1|2019-10-04|2021-04-08|Sederma|Use of leontodium alpinum plant cells for an anti-glycation anti-ageing skin treatment|
    WO2022023406A1|2020-07-30|2022-02-03|Sederma|Cosmetic or dermatological peptide-based treatment of the skin and its integuments|WO1998007744A1|1996-08-23|1998-02-26|Sederma S.A.|Synthetic peptides and their use in cosmetic or dermopharmaceutical compositions|
    FR2802769B1|1999-12-23|2004-01-16|Oreal|LEONTOPODIUM GENE PLANT EXTRACT AND COMPOSITIONS CONTAINING THE SAME|
    FR2821086B1|2001-02-21|2003-12-12|Sederma Sa|PROCESS FOR THE MANUFACTURE OF PROTEINS BY FERMENTATION OF MICROORGANISMS OF THE THERMUS FAMILY, MIXTURE OF PROTEINS THUS OBTAINED AND COSMETIC COMPOSITION CONTAINING THEM|
    FR2836042B1|2002-02-15|2004-04-02|Sederma Sa|COSMETIC OR DERMOPHARMACEUTICAL COMPOSITIONS FOR REDUCING POCKETS AND DARK CIRCLES UNDER EYES|
    FR2843023B1|2002-07-30|2004-09-24|Sederma Sa|COSMETIC OR DERMOPHARMACEUTICAL COMPOSITIONS CONTAINING KOMBUCHA.|
    FR2854897B1|2003-05-12|2007-05-04|Sederma Sa|COSMETIC OR DERMOPHARMACEUTICAL COMPOSITIONS FOR REDUCING THE SIGNS OF SKIN AGING.|
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    FR2869229B1|2004-04-26|2006-08-25|Sederma Soc Par Actions Simpli|USE OF A INDUCER OF UGT BY TOPIC|
    FR2941231B1|2009-01-16|2016-04-01|Sederma Sa|NOVEL PEPTIDES, COMPOSITIONS COMPRISING THEM AND COSMETIC AND DERMO-PHARMACEUTICAL USES|
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    FR2970868A1|2011-01-31|2012-08-03|Sederma Sa|PLANT ORIGIN EXTRACT, COMPOSITION CONTAINING THE SAME, PROCESS FOR OBTAINING BY VEGETABLE CULTURE AND USES IN THE COSMETIC, PHARMACEUTICAL AND COSMECEUTICAL FIELDS|
    FR2980362B1|2011-09-27|2013-10-04|Sederma Sa|NOVEL COSMETIC USE OF ALBIZIA JULIBRISSIN EXTRACT AND CORRESPONDING TOPICAL COMPOSITION|
    EP2623094A1|2012-02-02|2013-08-07|DSM IP Assets B.V.|Use of an edelweiss extract|CA3063229A1|2017-05-18|2018-11-22|Medena Ag|Use of naturally glycosylated polyphenols as protective agents against the effects of ultraviolet irradiation|
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    法律状态:
    2015-12-11| PLFP| Fee payment|Year of fee payment: 2 |
    2016-07-15| PLSC| Publication of the preliminary search report|Effective date: 20160715 |
    2016-12-19| PLFP| Fee payment|Year of fee payment: 3 |
    2017-12-18| PLFP| Fee payment|Year of fee payment: 4 |
    2019-12-20| PLFP| Fee payment|Year of fee payment: 6 |
    2020-12-10| PLFP| Fee payment|Year of fee payment: 7 |
    2022-01-19| PLFP| Fee payment|Year of fee payment: 8 |
    优先权:
    申请号 | 申请日 | 专利标题
    FR1550239A|FR3031454B1|2015-01-13|2015-01-13|USE OF LEONTOPODIUM ALPINUM VEGETABLE CELLS FOR COSMETIC TREATMENT AND CORRESPONDING ACTIVE COSMETIC INGREDIENT|
    FR1550239|2015-01-13|FR1550239A| FR3031454B1|2015-01-13|2015-01-13|USE OF LEONTOPODIUM ALPINUM VEGETABLE CELLS FOR COSMETIC TREATMENT AND CORRESPONDING ACTIVE COSMETIC INGREDIENT|
    EP16701200.4A| EP3244971A1|2015-01-13|2016-01-11|Use of plant cells of leontopodium alpinum for a cosmetic treatment and corresponding active ingredient|
    CN201680005652.1A| CN107106478A|2015-01-13|2016-01-11|The plant cell of edelweiss is used for the purposes and corresponding active component of cosmetic treatments|
    PCT/IB2016/050102| WO2016113659A1|2015-01-13|2016-01-11|Use of plant cells of leontopodium alpinum for a cosmetic treatment and corresponding active ingredient|
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