Water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa and its cosmetic uses.

Abstract

Cosmetic use of extracts of seeds and / or delipidated calyces of Hibiscus sabdariffa of the Malvaceae family with multifunctional effects and intended to be placed in contact with the various superficial parts of the human body.

Description

Title of the invention: Water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdanffa and its cosmetic uses.

[0001] The present invention relates to a cosmetic composition intended to be placed in contact with the various superficial parts of the human body (epidermis, hair and scalp, nails, lips and external genital organs) or with the teeth and oral mucosa, with a view exclusively or mainly to cleaning them, perfuming them, changing their appearance, protecting them, keeping them in good condition or correcting body odors, and characterized in that it comprises at least one aqueous, glycerinated or hydroalcoholic extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family, with the aim of advantageously utilizing a multifunctional active ingredient allowing: to reduce localized fat deposits, to stimulate the skin's natural antimicrobial defenses, its microbiota and microbiome, to promote its proper maturation and differentiation of the skin allowing it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, to activate the overall response of the skin to oxidative stress and to promote a better response to xenobiotics.

[0002] Xenobiotics are understood to mean the different categories of xenobiotics that the skin, the first barrier of our body, is able to detoxify and which include alcohol, medicines, drugs and endocrine disruptors as well as a very large number of products contained in the atmosphere or coming into contact with the skin surface such as pesticides, building materials or furniture and other industrial pollutants.

[0003] It is known that more and more cosmetics are trying to claim a more organic, more ecological, more natural mode of action with the new ISO 16128 standard, vegan, and even a slow cosmetics approach (ecological, healthy, intelligent, and reasonable) that takes into account the skin's real needs, advocating the use of fewer ingredients (skinimalism) to achieve better results, even frugality, positive contributions for the skin, and realistic promises. One response to this concern could then be plant extracts with multiple, multifunctional, and complementary properties that are sufficient on their own and therefore often do not require the addition of other active substances in cosmetic formulations.

[0004] In the context of the invention, a multifunctional active ingredient or multifunctional extract will be called an active ingredient or extract having several different and complementary efficacies at several levels of the skin structure in order to allow the creation of a cosmetic composition with the least amount of extract and active ingredient possible.

[0005] Many people believe that the skin surface to which cosmetics are applied is a dead and inert structure. In fact, this is not the case, and the epidermis, including its outermost stratum corneum, is the site of intense biochemical reactions where activators, inhibitors, and inhibitors of inhibitors maintain a balance and homeostasis that varies with the time of day, the seasons, and age. Each epidermal cell, and particularly the keratinocytes, which are the predominant cells of the epidermis, exhibits a complex and intricate metabolism leading to cell maturation and differentiation under the control of the expression of the 36,000 genes and variants that comprise them. The epidermis, located above the dermis and of ectodermal origin like the brain, can be compared to a conveyor belt. Keratinocytes divide by cell proliferation only at its base, then migrate towards the surface, progressively differentiating to form the stratum corneum, which forms the surface before normally desquamating imperceptibly. During this migration, a barrier is established, protecting the skin and the rest of the body from external aggressions and reducing transepidermal water loss.

[0006] It is then easy to imagine the interest of being able to select and test plant substances and extracts on the expression of all these genes in order to be able to select the most interesting extracts and develop the most efficient extraction and production methods.

[0007] Among all the plants studied, Hibiscus sabdariffa, from the Malvaceae family, has numerous and well-known properties when used as a whole, and particularly its oil. Indeed, the oil of this plant is traditionally used in West Africa to restore softness and elasticity to the skin, to relieve and soothe superficial burns, and is often applied to brittle, dry, dull, or split-ended hair to restore its shine and beauty.

[0008] This oil is produced from the seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family and its different varieties.

[0009] However, the use of oil is not always appreciated by users in cosmetics. Therefore, there is a need to provide other non-lipid captive forms of Hibiscus sabdariffa for both therapeutic and cosmetic purposes.

[0010] One of the aims of the invention is to provide a non-greasy composition.

[0011] Another object of the invention is to propose the use of such a composition in the context of a cosmetic application.

[0012] For the purposes of this invention, a seed and / or calyx extract shall be understood to mean an extract made from seeds and / or calyces, or an extract made exclusively from seeds, or an extract made exclusively from calyces. The calyces of the Hibiscus sabdariffa varieties referred to here correspond to the set of sepals that become fleshy—a remarkable botanical phenomenon—after the flower and seeds have separated from the plant. It is therefore important to distinguish between calyces and flowers, which are not botanically equivalent.

[0013] It was based on the observation that plants, and particularly their seeds, very often contain a significant amount of lipids that hinder the release of substances of interest and necessitate the use of complex solvents other than simple water, hydroethanolic solvents, or vegetable glycerin that the idea arose to separate these lipids beforehand from the rest of the plant to be extracted in order to facilitate the extraction of the non-lipid components of these seeds and / or calyces. Extraction is easier, yields are improved, and certain compounds are present that would not be present at all if this prior delipidation had not been undertaken.

[0014] It was by studying the properties of different water-soluble extracts of Hibiscus sabdariffa seeds after delipidation obtained after one or more successive cold pressings, on the expression of genes of normal human keratinocytes that the applicant discovered quite unexpectedly that these extracts of Hibiscus sabdariffa had many properties of interest to healthy skin and therefore to cosmetics particularly those concerned with minimalist formulas using multifunctional active ingredients since a given extract could have many complementary properties and reduce the use of many different active substances.

[0015] The invention therefore relates to a cosmetic composition comprising a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family and a cosmetically acceptable vehicle.

[0016] In the context of the invention, a water-soluble extract of a plant or part of a plant, such as seeds, is understood to be the result of any extraction in a water-soluble solvent, such as water itself, a hydroethanolic solution, or glycerin, and which can serve as a cosmetically acceptable vehicle and therefore be directly formulatable to produce a final cosmetic composition. These extracts can be formulated directly at concentrations of approximately 0.1% to 5% or stored in its dry state after evaporation of water or water plus ethanol or even in its current state in the case of a glycerinated solvent.

[0017] Also these cosmetic compositions are characterized in that said water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family is an extract from the seeds of one or more varieties of Hibiscus chosen from the group formed by Hibiscus sabdariffa white flower variety, Hibiscus sabdariffa vimto variety, Hibiscus sabdariffa koor red variety, Hibiscus sabdariffa CTL 92 variety, Hibiscus sabdariffa Thai variety, Hibiscus sabdariffa sabdariffa variety, Hibiscus sabdariffa altissima variety.

[0018] In the context of the invention and to facilitate its drafting, Hibiscus sabdariffa of the Malvaceae family shall therefore mean any of its different varieties, namely Hibiscus sabdariffa white flower variety, Hibiscus sabdariffa vimto variety, Hibiscus sabdariffa koor red variety, Hibiscus sabdariffa CTL 92 variety, Hibiscus sabdariffa Thai variety, Hibiscus sabdariffa sabdariffa variety, Hibiscus sabdariffa altissima variety.

[0019] Therefore, a cosmetic or pharmaceutical composition intended to be placed in contact with the various superficial parts of the human body (epidermis, hair and scalp, nails, lips and external genital organs) or with the teeth and oral mucosa, with a view exclusively or mainly to cleaning them, perfuming them, changing their appearance, protecting them, keeping them in good condition or correcting body odors, and characterized in that it comprises at least one water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family, with the aim of advantageously utilizing a multifunctional active ingredient that allows both: to reduce localized fat deposits, to stimulate the skin's natural antimicrobial defenses, its microbiota and microbiome, to promote its proper maturation and differentiation of the skin allowing it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, to activate the overall response of the skin to oxidative stress and to promote a better response to xenobiotics.

[0020] The applicant has indeed tested several aqueous extracts with or without enzymes, or hydroethanolic extracts of Hibiscus sabdariffasw, the complete genome of normal human keratinocytes, and particularly by genome-wide gene expression analysis (36,000 transcribed genes and variants) of normal human epidermal keratinocytes following the extraction of total transcribed Ribonucleic Acids (RNA) followed by Deoxyribonucleic Acid microarray hybridization (DNA). The principle of the DNA microarray is based on the property of denatured DNA (single strand) to spontaneously reform its double helix when in the presence of a complementary strand (hybridization reaction). Since it is possible to attach up to one million probes to a biochip, DNA microarrays thus constitute a mass-market approach that the inventor has used, as they allow, in a single experiment, for an estimation of the expression of tens of thousands of genes.

[0021] Advantageously, said pharmaceutically acceptable vehicle is also that used as solvent during extraction and is chosen from water with or without enzyme, a hydroethanolic solution or a water / glycerol mixture.

[0022] Cosmetic compositions according to claims 1 and 2 characterized in that the water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family is an extract from the seeds and / or calyces of one or more varieties of Hibiscus selected from the group formed by Hibiscus sabdariffa white flower variety, Hibiscus sabdariffa vimto variety, Hibiscus sabdariffa koor red variety, Hibiscus sabdariffa CTL 92 variety, Hibiscus sabdariffa Thai variety, Hibiscus sabdariffa sabdariffa variety, Hibiscus sabdariffa altissima variety.

[0023] Also advantageously, said water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family is made from seeds and / or calyces previously delipidated by cold pressing, one to three times successively, in order to obtain a content of 10% or less by mass of lipids in relation to the total mass of the seeds and / or calyces so that in the end the extract comprises less than 1% by mass of lipids in relation to the total mass of the extract, in particular from 0% to 1% by mass of lipids in relation to the total mass of the extract.

[0024] In the context of the invention, "from 0% to 1% by mass of lipids" means that the extract may comprise 0, 0.25, 0.5, 0.75, 1% by mass of lipid relative to the mass of the extract.

[0025] Also within the scope of the invention, said extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family has been made water-soluble by solubilization in a solvent chosen from water with or without enzyme, a hydroethanolic solution or a water / glycerol mixture before being able to be used as is in the composition or stored after complete drying before being resolubilized in these same cosmetically acceptable solvents before use.

[0026] Advantageously, the cosmetic composition is characterized in that said water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family represents from 0.1% to 5% by mass relative to the total mass of the composition.

[0027] In the context of the invention, "from 0.1% to 5% by mass" means that the composition may comprise 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5% by mass of delipidated extract relative to the total mass of the composition.

[0028] The effects of different plant extracts on gene expression in Normal Human Epidermal Keratinocytes (NHEK) were investigated. More specifically, a full transcriptome analysis, i.e., the analysis of all genes expressed at a given time, was performed using the Affymetrix GeneAtlas platform and the U219 "full human transcriptome" chip containing 36,000 so-called transcribed genes and variants.

[0029] This method, described in more detail in the examples below, allows us to choose, based on the genes stimulated, i.e. whose expression is significantly increased, or inhibited, i.e. whose expression is significantly reduced, the most interesting plants for cosmetic use and which has shown the interest of Hibiscus sabdariffa from the Malvaceae family among other plants thus tested.

[0030] Then, by taking the most interesting gene or genes, this method makes it possible to select the optimum protocol(s) in order to obtain the best possible results. Here, for Hibiscus sabdariffa of the Malvaceae family, the ANGPTL4 gene for ANGgioPoietin-Like 4 is involved in lipid metabolism as an inhibitor of lipoprotein lipase and pancreatic lipase. Lipoprotein lipase is a key enzyme in the penetration of lipids into adipocytes in adipose tissue and is responsible for localized fat deposits that many cosmetics seek to reduce through topical application. This gene was used to determine that it was the delipidated seeds and / or calyces extracted in a simple solvent that resulted in an increase in gene expression of more than 1212% upon contact with normal human keratinocytes (NHEK). The relative expression of this ANGPTL4 gene in control keratinocytes had a signal intensity of 30.25, while in keratinocytes exposed to the thus-selected water-soluble extract of Hibiscus sabdariffa, the signal intensity was 396.90, representing a 13.12-fold increase in gene expression, known as a fold change, under the experimental conditions.

[0031] This made it possible to determine an optimum when the extraction was carried out in simple solvents such as water, hydroethanolic mixtures in which the ethanol is then recycled after complete evaporation or mixtures of water and glycerin, whose moisturizing properties for the skin are well established. These tests demonstrate the value of a cosmetic composition characterized by its composition of a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa from the Malvaceae family. This extract is obtained by mechanically separating the seeds from the fruits, which are then ground and cold-pressed several times successively, with or without prior roasting. This is followed by oven drying for 1 to 5 hours at a temperature between 50°C and 60°C to prevent the proliferation of microorganisms. The product is then extracted or macerated in water with or without added enzymes, a hydroethanolic solution, a water / glycerol mixture, or macerated in glycerol alone. It can then be filtered or centrifuged and stored and used directly in its respective solvent, or dried again, freeze-dried, or even zeodrated, thus enabling...Using a single multifunctional extract: to reduce localized fat deposits, to stimulate the skin's natural antimicrobial defenses, its microbiota and microbiome, to promote proper skin maturation and differentiation, resulting in a more beautiful surface, greater resistance to the harmful effects of external aggressions and aging due to a superior barrier effect, to activate the skin's overall response to oxidative stress, and to promote a better response to xenobiotics.

[0032] One way according to the invention to reduce localized weight gain, also called cellulite, using a single multifunctional extract is to activate the expression of the genes for Angiopoietin (ANGPTL4), Perilipin 2 (PLIN2), GO / G1 Switch (GOS2), Camitin palmitoyl transferase (CPT1A); Indeed, ANGPTL4 inhibits lipoprotein lipase and pancreatic lipase, the latter promoting the storage of fatty acids in adipocytes, which is the cause of the so-called cellulite. PLIN2, perilipin or adipophilin ensures the maintenance of adipose tissue. GOS2 adds a circadian effect to fat storage and is highly present in subcutaneous adipose tissue. And CPT1A promotes lipid degradation in mitochondria.

[0033] According to the invention, a way to stimulate the skin's natural antimicrobial defenses, its microbiota and its microbiome, using a single multifunctional extract, is to activate the expression of antimicrobial peptides and their genes (S100A7, 8, 9) and that of Defensin B1 (DEFB1). Indeed, the S100 proteins (S100A7, S100A8 and S100A9) as well as DEFB1 are antimicrobial proteins and peptides, both bacterial and fungal, ensuring the innate immune defense of the skin and effectively regulating the cutaneous microbiota and its microbiome.

[0034] According to the invention, one way to promote, with the help of a single multifunctional extract, the proper maturation and differentiation of the skin, allowing it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, is to stimulate the expression of the genes of Claudin 4 (CLDN4), Occludin (OCLN), Small Proline Rich Proteins 2A (SPRR2A), Sciellin (SCEL), Epiregulin (EREG), Serpine B1 and B2 (SERPIN B1 and B2) and to reduce the expression of the genes of Matrix Metalloproteinases 1 and 10 (MMP3, MMP10) and Chemokine 14 (CXCL14). Indeed, CLDNA codes for Claudin4, an essential component of tight junctions in the deep epidermis, which plays an important role in the skin's barrier effect and the control of its insensible water loss. Similarly, OCLN codes for Occludin, which is also located in these tight junctions, ensuring their formation, maintenance, and functions. PPRR2, EREG, Serpines G1 and G2 and SCEL, participate in the terminal differentiation of the epidermis and allow the stratum corneum and the corneal envelopes to finalize their overall barrier effect and protection against aggressions of all types. The reduction of the expression of matrix metaloproteinases such as MMP3 and MMP10, which in some situations destroy the extracellular matrix, and of CXCL14, a pro-inflammatory chemokine, contributes to maintaining this functional barrier effect in association with the antiproteinase effect of Serpins B1 and B2, which are activated elsewhere as described above.

[0035] A way according to the invention of activating the overall response of the skin to oxidative stresses and enabling it to have a better response to xenobiotics, using a single multifunctional extract, is to activate the so-called Nrf2 signaling pathway which, when activated, plays a cytoprotective role against oxidative stresses and those arising from certain exposures to xenobiotics. This cytoprotective signaling pathway is particularly activated by stimulation of the expression of cytochrome P450 (CYP1B1), Aldo-ketoreductase 1 AKR1C1, superoxide dismutase 2 (SOD2) and Insulin-like growth factor binding protein 3 (IGFBP3) genes, which together play a cytoprotective role.

[0036] According to the invention, said water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffade from the Malvaceae family strongly reduces the expression of the WNT4 gene which inhibits hair regrowth and would make it an extract favorable to regrowth at the beginning of baldness.

[0037] The invention also relates to a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family, characterized in that it includes water with or without enzyme, a hydroethanolic solution or a water / glycerol mixture and is prepared from 10% by weight of delipidated and then dried seeds and / or calyces.

[0038] This allows obtaining a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family, characterized in that it is intended to be applied to healthy skin to strengthen and promote on the one hand the reduction of localized fat deposits and on the other hand: stimulate the skin's natural antimicrobial defenses, its microbiota and its microbiome, promote its proper maturation and differentiation, allowing it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, activate the overall response of the skin to oxidative stress and promote a better response to xenobiotics.

[0039] Or, alternatively, a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family for the treatment or prevention of the negative effects caused by xenobiotics, molecules foreign to the body that often require detoxification, oxidative and cellular stress, microorganisms, their microbiota and microbiome, localized fat overload, oxygen free radicals, and the slowing of proper keratinocyte maturation and differentiation, all linked to an under-expression of Angiopoietin (ANGPTL4), Perilipin 2 (PLIN2), GO / G1 Switch (GOS2), Carnitine palmitoyltransferase (CPT1A), antimicrobial peptides (S100A7, 8, and 9), Defensin B1 (DEFB1), Claudin 4 (CLDN4), and Occludin (OCLN), Small Proline Rich Proteins 2A (SPRR2A), sciellin (SCEL), Epiregulin (EREG), Serpin B1 and B2 (SERPIN B1 and B2), cytochrome P450 (CYP1B1),of Aldo-ketoreductase 1 AKR1C1, superoxide dismutase 2 (SOD2) and insulin-like growth factor binding protein 3 (IGFBP3), or, conversely, in the context of the overexpression of genes promoting skin problems, the onset of baldness, cell hyperproliferation following UV irradiation or its inflammation, such as matrix metalloproteinases 1 and 10 (MMP3, MMP10), chemokine 14 (CXCL14) and wingless type 4 (WNT4).

[0040] Advantageously, the invention relates to the non-therapeutic cosmetic use of a composition for topical application to healthy skin, with the aim of advantageously utilizing a multifunctional extract that simultaneously reduces localized fat deposits, stimulates the skin's natural antimicrobial defenses, microbiota, and microbiome, and promotes its proper maturation and differentiation, thus making it more beautiful on the surface and more resilient. to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, to activate the overall response of the skin to oxidative stress and to promote a better response to xenobiotics.

[0041] For cosmetic use, the interest then becomes apparent in a composition comprising a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family, characterized in that the composition is in the form of an oil-in-water (O / W) or water-in-oil (W / O) emulsion, a multiple emulsion (O / W / W, W / O / W), a microemulsion, a twin-phase emulsion, a PIT emulsion, a nanoemulsion, a pseudoemulsion, an aqueous gel, a fatty gel, a hydroalcoholic gel, a suspension, a lyophilized gel or emulsion, or one of the preceding forms, containing microcapsules, nanocapsules, liposomes, ethosomes, and which can be presented as creams, serums, liquids, pastes, lotions, emulsions, gels, solids, powders, masks, sticks, sprays, aerosols.

[0042] As well as a composition comprising a water-soluble extract of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family characterized in that it is in the form of a two-phase micellar makeup remover, a foaming gel, a jelly lotion, an exfoliant, a moisturizing cream, a moisturizing serum, a wrinkle-enhancing serum, a moisturizing body lotion, a regenerating mask, a regenerating cream, a slimming body cream, an anti-aging cream, a regenerating serum, a balm, a mist, an eye treatment, a concentrate, or a detoxifying mask.

[0043] A cosmetic skincare method for healthy skin to advantageously utilize a multifunctional active ingredient that simultaneously reduces localized fatty deposits, stimulates the skin's natural antimicrobial defenses, its microbiota and microbiome, promotes proper skin maturation and differentiation, resulting in a more beautiful surface, greater resistance to the harmful effects of external aggressions and aging due to a superior barrier effect.to activate the skin's overall response to oxidative stress and to promote a better response to xenobiotics, including the topical administration of a cosmetic composition of a water-soluble extract of delipidated Hibiscus sabdariffa seeds and / or calyces, optimized by the use of whole-genome expression analysis of cultured human keratinocytes (NHEK) in order to determine all its properties as well as the optimization of its preparations, the association of enzymes or not during the extraction or maceration steps, the benefit or not of prior roasting, the percentage of delipidated and dried seeds and / or calyces or the nature of the final drying and characterized both by an increase in the expression of certain genes of interest and by a decrease in the expression of other genes of interest.

[0044] Examples are then given without being limiting in order to illustrate the invention with two different extraction situations, the first in examples 1 and 2 with just water as a solvent and the second in examples 3 and 4 where 5% of enzymes have been added to the water used as a solvent.

[0045] Example 1. Obtaining delipidated water-soluble extracts of seeds and / or calyces of Hibiscus sabdariffa in a pure water solvent. Extracts of delipidated seeds and / or calyces of Hibiscus sabdariffa of the Malvaceae family are made by mechanical separation of the seeds from the fruits followed by a minimum drying of 1 day and then by one or more cold pressings of these seeds, after or without prior roasting, followed by drying in an oven for 1 to 5 hours and at a temperature between 50°C and 60°C in order to avoid the proliferation of microorganisms, then by extraction or maceration in water or again dried or freeze-dried or even zeodrated. It therefore includes pure water and is prepared from 10% by weight of delipidated and then dried seeds and / or calyces. These extracts can be tested directly or they can be dried, dehydrated, freeze-dried or zeodrated before testing. In the latter cases they are rehydrated or diluted with purified water acting as a solvent. We will call the extracts obtained and used in the following examples EHS (for Hibiscus Sabdariffa Extract).

[0046] Example 2. Tests and examples of results. The biological model: Third-pass normal human epidermal keratinocytes (NHEK) cultured at 37°C and 5% CO2. Culture medium supplemented with EGF at 0.25 ng / ml and EP at 25 µg plus gentamicin 25 µg. The test medium consisted of SFM keratinocytes with only gentamicin 25 µg / ml. Experimental conditions: Keratinocytes were seeded in 24-well plates and cultured in culture medium for 24 hours, then in test medium for an additional 24 hours. The medium was then replaced with test medium containing or not (control) the compounds under investigation, and the cells were incubated for 24 hours. All experimental conditions were performed in triplicate (n=3). At the end of incubation, the culture supernatants were removed and the cell mats were rinsed with a PBS solution. The plate was immediately frozen dry at -80°C. Prior to extraction, the culture replicates were pooled. Total RNA from each sample was extracted using TriPure Isolation Reagent® according to the protocol recommended by the supplier. The quantity and quality of RNA were assessed by capillary electrophoresis (Bioanalyzer 2100, Agilent). The synthesis of biotinylated antisense RNAs (aRNAs) was carried out using the "GeneChip 3'IVT Express" kit (Affymetrix®). For each biotinylated RNA sample, an electrophoretic profile was performed (Bioanalyzer 2100, Agilent) before and after fragmentation. The hybridization of labeled and fragmented RNAs on the Affymetrix® U219 chip (36,000 transcripts and variants) was carried out on the GeneAtlas™ fluidics Affymetrix ® hybridization station for 20 hours at 45 °C. The U219 chips were then scanned using the GeneAtlas™ Imaging station (Affymetrix® - 2 pm resolution) to generate signal intensity data as shown in the result examples below. Signal intensity data were normalized using Expression Console software (Affymetrix®), based on the RMA algorithm. Quality control of the labeling and hybridization was then performed. Quality controls of the hybridization and labeling steps validated the experimental process Once normalized with Expression Console software, the data were transferred and formatted in a Microsoft Excel file. Calculations and filters have been added to these files to sort the data and facilitate its use. The "fold change" thresholds or rate of change (value corresponding to the ratio: signal intensity value of a probe corresponding to the treated sample / signal intensity value of a probe corresponding to the control) were defined and applied to the normalized data. The following abbreviations are used in the analysis files: - UR: Upregulated - overexpression or stimulation of the probe or gene when the fold change is greater than 2, - DR: Downregulated - underexpression or inhibition of the probe or gene when the fold change is less than 0.5. Each analysis file consists of: - The signal intensity value (RE: Relative Expression) corresponding to each of the samples, - Calculations of "fold change" or rate of change for each comparison, - Information relating to each gene. The filters inserted into these files allow the selection of significantly modulated genes. The example of gene probes corresponding to the variation in expression of ANGPTL4, CLDN4, PLIN2, S100A7, OCLN, SERPIN B2, IGFBP3, GOS2, EREG, CPT1A, DEFB1, SCEL and of MMP3, CXCL14, WNT4 thus obtained with this methodology and with EHS for the extract of delipidated seeds and / or calyces of Hibiscus Sabdariffa, the numbers indicated correspond to the intensity of the signal obtained for the control and for the EHS assay: Signal intensity for Angiopoietin 4 (ANGPTL4): - Witness: 30.65 - EHS: 396.90 - Change rate: 13.12 - Regulation: UR Signal intensity for Claudine 4 (CLDN4): - Witness: 42.57 - EHS: 387.44 - Change rate: 9.10 - Regulation: UR Signal intensity for Perilipin 2 (PLIN2): - Witness: 85.98 - EHS: 475.20 - Change rate: 5.53 - Regulation: UR Signal intensity for Calcium Binding Protein A7 (S100A7): - Witness: 33.99 - EHS: 133.36 - Change rate: 4.17 - Regulation: UR Signal intensity for Occludine (OCLN): - Witness: 86.51 - EHS: 320.06 - Change rate: 3.70 - Regulation: UR Signal intensity for Serine protease B2 (Serpine B2): - Witness: 417.19 - EHS: 856.55 - Change rate: 2.05 - Regulation: UR Signal intensity for Insulin-like Growth Factor Binding Protein 3 (IGFBP3): - Control: 40.00 -EHS: 119.06 - Change rate: 2.98 - Regulation: UR Signal strength for the GO / G1 (GOS2) switch: - Witness: 20.00 - EHS: 56.77 - Change rate: 2.84 - Regulation: UR Signal intensity for Epiregulin (EREG): - Witness: 167.00 -EHS: 451.53 - Change rate: 2.70 - Regulation: UR Signal intensity for Camitine palmitoyltransferase (CPT1A): - Witness: 351.39 - EHS: 887.17 - Change rate: 2.52 - Regulation: UR Signal intensity for Defensin (DEFB1): - Witness: 42.76 - EHS: 89.76 - Change rate: 2.10 - Regulation: UR Signal intensity for the Scielline (SCEL): - Witness: 65.00 -EHS: 118.91 - Change rate: 1.83 - Regulation: UR Signal intensity for Metalloproteinase 3, MMP3: - Witness: 290.23 - EHS: 37.05 - Change rate: 0.13 - Regulation: DR Signal intensity for Chemokine 14 (CXCL14): - Witness: 2113.61 - EHS: 356.24 - Change rate: 0.17 - Regulation: DR Signal intensity for Wingless type 4 (WNT4): - Indicator: 99.42 - EHS: 27.09 - Change rate: 0.27 - Regulation: DR

[0047] Example 3. Obtaining delipidated water-soluble extracts of Hibiscus sabdariffa seeds and / or calyces in a water solvent containing 5% enzymes. The delipidated extracts of Hibiscus sabdariffa seeds and / or calyces from the Malvaceae family are obtained by mechanical separation of the seeds from the fruits followed by a minimum drying of 1 day and then by one or more cold pressings of these seeds, after or without prior roasting, followed by oven drying for 1 to 5 hours at a temperature between 50°C and 60°C in order to avoid the proliferation of microorganisms, then by extraction or maceration in water with enzymes, or again dried or freeze-dried or even zeodrated. It therefore includes water with 5% enzymes and is prepared from 10% by weight of delipidated and then dried seeds and / or calyces. These extracts can be tested directly or they can be dried, dehydrated, freeze-dried or zeodrated before testing. In the latter cases they are rehydrated or diluted with purified water acting as a solvent. A control was also prepared containing water and 5% of these same Enzymes in order to serve as a control (control E). We will call the extracts obtained and used in the following examples EHS (for Hibiscus Sabdariffa Extract).

[0048] Example 4. Tests and examples of results. The biological model: Third-pass normal human epidermal keratinocytes (NHEK) cultured at 37°C and 5% CO2. Culture medium supplemented with EGF at 0.25 ng / ml and EP at 25 µg plus gentamicin 25 µg. The test medium consisted of SFM keratinocytes with only gentamicin 25 µg / ml. Experimental conditions: Keratinocytes were seeded in 24-well plates and cultured in culture medium for 24 hours, then in test medium for an additional 24 hours. The medium was then replaced with test medium containing or not (control) the compounds under investigation, and the cells were incubated for 24 hours. All experimental conditions were performed in triplicate (n=3). At the end of incubation, the culture supernatants were removed and the cell mats were rinsed with a PBS solution. The plate was immediately frozen dry at -80°C. Prior to extraction, the culture replicates were pooled. Total RNA from each sample was extracted using TriPure Isolation Reagent® according to the protocol recommended by the supplier. The quantity and quality of RNA were assessed by capillary electrophoresis (Bioanalyzer 2100, Agilent). The synthesis of biotinylated antisense RNAs (aRNAs) was carried out using the "GeneChip 3'IVT Express" kit (Affymetrix®). For each biotinylated RNA sample, an electrophoretic profile was performed (Bioanalyzer 2100, Agilent) before and after fragmentation. The hybridization of labeled and fragmented RNAs on the Affymetrix® U219 chip (36,000 transcripts and variants) was carried out on the GeneAtlas™ fluidics Affymetrix ® hybridization station for 20 hours at 45 °C. The U219 chips were then scanned using the GeneAtlas™ Imaging station (Affymetrix® - 2 pm resolution) to generate signal intensity data as shown in the result examples below. Signal intensity data were normalized using Expression Console software (Affymetrix®), based on the RMA algorithm. Quality control of the labeling and hybridization was then performed. Quality controls of the hybridization and labeling steps validated the experimental process. Once normalized with Expression Console software, the data were transferred and formatted in a Microsoft Excel file. Calculations and filters have been added to these files to sort the data and facilitate its use. The "fold change" thresholds or rate of change (value corresponding to the ratio: signal intensity value of a probe corresponding to the treated sample / signal intensity value of the probes corresponding to the signals of the mixture of enzymes alone serving as control, controls E) were defined and applied to the normalized data. The following abbreviations are used in the analysis files: - UR: Upregulated - overexpression or stimulation of the probe or gene when the fold change is greater than 2, - DR: Downregulated - underexpression or inhibition of the probe or gene when the fold change is less than 0.5. Each analysis file consists of: - The signal intensity value (RE: Relative Expression) corresponding to each of the samples, - Calculations of "fold change" or rate of change for each comparison, - Information relating to each gene. The filters inserted into these files allow the selection of significantly modulated genes. The example of gene probes corresponding to the variation in expression of CYP1B1, IGFBP3, AKR1C1, S100A8, SERPIN Blet B, SOD2, S100A8, SPRR2A, EREG and MMP 10, thus obtained with this methodology and with EHS for the extract of delipidated seeds and / or calyces of Hibiscus Sabdariffa, the numbers indicated correspond to the intensity of the signal obtained for the control E and for the EHS assay: Signal intensity for Cytochrome P450 (CYP1B1): - Witness E: 40.00 - EHS: 484.65 - Change rate: 12.12 - Regulation: UR Signal intensity for Insulin-like Growth Factor Binding Protein 3 (IGFBP3): - Witness E: 40.00 - EHS: 356.26 - Change rate: 6.41 - Regulation: UR Signal intensity for Aldo-ketoreductase 1 (AKR1C1): - Witness E: 214 - EHS: 1269 - Change rate: 5.94 - Regulation: UR Signal intensity for Small Protein Rich in Proline (SPRR2A): - Witness E: 53.48 - EHS: 150.09 - Change rate: 2.81 - Regulation: UR Signal intensity for Calcium Binding Protein A8 (S100A8): - Witness E: 509.76 - EHS: 1556.79 - Change rate: 3.05 - Regulation: UR Signal intensity for Serine protease B1 (Serpin Bl): - Witness E: 70.21 - EHS: 237.73 - Change rate: 3.39 - Regulation: UR Signal intensity for Superoxide Dismutase 2 (SOD2): - Witness E: 403.63 - EHS: 1268.52 - Change rate: 3.14 - Regulation: UR Signal intensity for Serine protease B2 (Serpin B2): - Witness E: 478.05 - EHS: 1018.98 - Change rate: 2.13 - Regulation: UR Signal intensity for Epiregulin (EREG): - Witness E: 186.51 - EHS: 322.23 - Change rate: 1.73 - Regulation: UR Signal intensity for Matrix Metalloproteinase 10 (MMP 10): - Witness E: 139.71 -EHS: 41.71 - Change rate: 0.30 - Regulation: DR

[0049] The presence of enzymes in the water of the extraction medium allows the genes of the Nrf2 signaling pathway to be expressed, for example the genes CYP1B1, AKR1C1 and SOD2, whereas extraction with pure water favors the expression of genes involved in epidermal differentiation and its barrier effect, such as CLDN4, OCLN, DEFB and SCEL. The expression of metalloproteinases is partially inhibited in both situations.

[0050] In the examples below, which give examples of the realization of these compositions, without this list being exhaustive, the following abbreviations will be used: EHS for the extract of delipidated seeds and / or calyces of Hibiscus Sabdariffa and Hibiscus Oil for the oil of seeds and / or calyces of the corresponding variety.

[0051] Examples. According to a first embodiment of the invention, the composition comprises: Carboxypolymethylene or carboxylic polymer..................0.40 g Diethanolamine cetylphosphate............................................1.50g Octyl dodecanol............................................................5.50 g Cetyl alcohol................................................................1.00 g Mineral oil...............................................................2.00 g Stearic acid..............................................................1.00 g Sterylic alcohol ethoxylate (20 E).........................................0.30 g Ketostearyl alcohol....................................................1.80 g Preservative......................................................................0.15 Hibiscus Oil..........................................................0.50 g Triethanolamine..........................................................0.60 g Cyclomethicone.............................................................12.00 g Dimethicone copolyol........................................................3.00 g Phenyl trimeticone...........................................................2.00 g EHS........................................................................3.00 g Dextran........................................................................1.00 g Glycerin..........................................................................2.00 g Demineralized water qsp........................................................100 g After 4 weeks of use on 25 volunteers, skin appeared smoother just 2 hours after each application. At the end of the treatment, the number, length, and width of wrinkles had significantly decreased, and 92% of users reported more hydrated, more beautiful, and less stressed skin.

[0052] Example 6. According to a second embodiment of the invention, the composition comprises: - Acrylate C10-30........................................................0.50 g - Arginine............................................................0.001g - Glycerin..................................................................................1.00 g - EHS..........................................................................................0.20 g - Alcohol..................................................................2.00 g - Carbomer................................................................................0.126 g - EDTA..............................................................................................0.005 g - Preservative......................................................................0.867 g - Hibiscus Oil.........................................................................0.5 g - Hydrogenated lecithin.....................................................0.09 g - Hyaluronic acid....................................................................0.20 g - TEA 99%...................................................................................0.62 g - Colorings........................................................................0.0002 g - Demineralized water qsp.................................................................100 g After 4 weeks of use for 20 volunteers aged 20 to 70, facial skin appeared significantly more hydrated and was like purified and more resistant to occasional infections and dehydration.

[0053] Example 7. Oil-in-water emulsion: - Liquid paraffin....................................................6 g - Liquid lanolin................................................................3 g - Arlacel 165................................................................6 g - Tween 60...................................................................2 g - Cetyl alcohol................................................................1.2 g - Stearic acid.................................................2.5 g - Arginine........................................................................0.5 g - Volatile silicone oil...................................................10 g - Triethanolamine................................................................0.1 g - Preservative....................................................................0.3 g - EHS........................................................................5 g - Hyaluronic acid.......................................................0.5 g - Demineralized water qsp.....................................................100 g After 4 weeks of twice-daily use for 20 volunteers aged 33 to 50 years, localized excess weight in the saddlebag area was significantly reduced.

[0054] Example 8. Liposome cream: -Cetyl alcohol.............................................................................4 g -B-sitosterol...................................................................................4 g -Dicetyl phosphate..................................................................................0.5 g -Preservative..............................................................................0.3 g -Carbopol 981 ...................................................................0.2 g -Triethanolamine............................................................................0.2 g -Phospholipid.............................................................................0.05 g -EHS..................................................................................1.5g -Demineralized water qsp................................................................100 g After use on a panel of 30 volunteers aged 18 to 70, the skin appeared significantly hydrated during the 3 hours of the first application, softer and younger-looking for 85% of users after 4 weeks of daily use.

[0055] Example 9. Water / Oil Emulsion: - Protein.......................................................................19g - Glycerin....................................................................................3 g - Vaseline oil.........................................................................8 g - Mg sulfate.............................................................................0.5 g -EHS...................................................................................0.5g - Hyaluronic acid.................................................................0.02 g - Preservative...........................................................2 g - Demineralized water qsp.............................................................100 g

[0056] Example 10. O / W gel emulsion - Carbopol 981 ..............................................................0.6 g - Ethyl alcohol..........................................................................15 g - Volatile silicone oil...............................................................3 g - Hibiscus Oil..............................................................................7 g - Preservative....................................................................0.3 g - Perfume........................................................................0.4 g -EHS........................................................................................0.1g - Triethanolamine..............................................................................0.2 g - Demineralized water qsp....................................................................100 g

[0057] Example 11. Glycerin concentrate -Glycerin...........................................................65 g -EHS..........................................................................................5 g - Arginine............................................................................1 g - Hyaluronic acid................................................ 1 g - Perfume........................................................0.4 g - Demineralized water qsp......................................................100 g

Claims

Demands

1. Cosmetic composition comprising a water-soluble seed extract obtained by cold pressing one to three times successively, of Hibiscus sabdariffa of the Malvaceae family representing 0.1% to 5% by mass relative to the total mass of this composition and a cosmetically acceptable vehicle which advantageously may be that used as a solvent during the extraction and is chosen from water with or without enzyme, a hydroethanolic solution or a water / glycerol mixture.

2. Cosmetic composition according to claim 1, wherein said cosmetically acceptable vehicle is also that used as solvent during extraction and is selected from water with or without enzyme, a hydroethanolic solution or a water / glycerol mixture.

3. Cosmetic compositions according to claims 1 and 2 characterized in that said water-soluble extract of delipidated Hibiscus sabdariffa seeds of the Malvaceae family is an extract from the seeds of one or more varieties of Hibiscus selected from the group formed by Hibiscus sabdariffa white flower variety, Hibiscus sabdariffa vimto variety, Hibiscus sabdariffa koor red variety, Hibiscus sabdariffa CTL 92 variety, Hibiscus sabdariffa Thai variety, Hibiscus sabdariffa sabdariffa variety, Hibiscus sabdariffa altissima variety.

4. Cosmetic composition according to any one of claims 1 to 3 characterized in that said water-soluble extract of delipidated Hibiscus sabdariffa seeds of the Malvaceae family is made from seeds previously delipidated by cold pressing, one to three times successively, in order to obtain a content of 10% or less by mass of lipids relative to the total mass of the seeds so that in the end the extract comprises less than 1% by mass of lipids relative to the total mass of the extract, in particular from 0% to 1% by mass of lipids relative to the total mass of the extract.

5. A cosmetic composition according to any one of claims 1 to 4, characterized in that said defatted seed extract of Hibiscus sabdariffa of the Malvaceae family has been rendered water-soluble by solubilization in a solvent selected from water with or without an enzyme, a hydroethanolic solution, or a water / glycerol mixture before being usable as is in the composition or stored after complete drying before being resolubilized in these same cosmetically acceptable solvents prior to use.

6. Composition according to any one of claims 1 to 5 characterized in that said water-soluble extract of delipidated Hibiscus sabdariffa seeds of the Malvaceae family represents from 0.1% to 5% by mass relative to the total mass of the composition.

7. Composition according to any one of claims 1 to 6, wherein the composition is in the form of an oil-in-water (O / W) or water-in-oil (W / O) emulsion, multiple emulsion (O / W / O, W / O / O), microemulsion, twin-phase emulsion, PIT emulsion, nanoemulsion, pseudoemulsion, aqueous gel, oily gel, hydroalcoholic gel, suspension, lyophilized gel or emulsion or any of the preceding forms containing microcapsules, nanocapsules, liposomes, ethosomes and may be in the form of creams, serums, liquids, pastes, lotions, emulsions, gels, solids, powders, masks, sticks, sprays, aerosols.

8. Composition according to any one of claims 1 to 6, said composition being in the form of a two-phase micellar makeup remover, a foaming gel, a jelly lotion, an exfoliant, a moisturizing cream, a hydrating serum, a wrinkle-enhancing serum, a moisturizing body lotion, a regenerating mask, a regenerating cream, a slimming body cream, an anti-aging cream, a regenerating serum, a balm, a mist, an eye treatment, a concentrate, or a detoxifying mask.

9. Non-therapeutic cosmetic use of a composition as defined in any one of claims 1 to 8 for the purpose of advantageously using a multifunctional extract that allows both the reduction of localized fat deposits, the stimulation of its natural antimicrobial defenses, its microbiota and its microbiome, the promotion of its proper maturation and differentiation, thus enabling it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because it presents a better quality barrier effect, the activation of the skin's overall response to oxidative stress and the promotion of a better response to xenobiotics.

10. Composition according to any one of claims 1 to 8 to act significantly, using a single multifunctional extract, on the proper maturation and differentiation of the skin, allowing it to be more beautiful on the surface, more resistant to the harmful effects of external aggressions and aging because presenting a better quality barrier effect is, thanks to these water-soluble extracts of Hibiscus sabdariffa seeds, to stimulate the expression of the genes of Claudin 4 (CLDN4), Occludin (OCLN), Small Proline Rich Proteins 2A (SPRR2A), Sciellin (SCEL), G0 / G1 Switch (GOS2), Epiregulin (EREG), Serpine B1 and B2 (SERPIN B1 and B2) and to reduce the expression of the genes of Matrix Metalloproteinases 1 and 10 (MMP3, MMP10) and Chemokine 14 (CXCL14).