Anhydrous extract of stem-free leaves of Hippophae rhamnoides or a composition containing said extract for use in maintaining and / or improving skin microcirculation

Abstract

The use of an anhydrous extract of stem-free leaves of Hippophae rhamnoides or a cosmetic composition comprising said extract for use in reducing dark circles and / or hollows in the eye contour and / or in maintaining and / or providing a lighter skin tone is disclosed.

Description

[Technical Field]

[0001] The present invention relates to the cosmetic and / or dermatological use of an anhydrous liquid extract of stemless leaves of Hippophae rhamnoides or a cosmetic and / or dermatological composition comprising said extract for maintaining and / or improving microcirculation in the skin. [Background technology]

[0002] The under-eye area, the zone bordering the lower eyelid above and the cheekbone below, is of greatest concern in cosmetics worldwide, as it is the first zone on the face to show signs of fatigue and stress.

[0003] Anatomically, the skin under the eyes is very thin, approximately 0.5 mm thick (one-third the thickness of the rest of the face), and is therefore extremely sensitive to environmental stressors (UV, air pollution, alcohol, stress, lack of sleep). As a result, the skin under the orbit is highly susceptible to free radical attack and can easily become inflamed.

[0004] Thus, the surrounding darkening and swelling, which corresponds to localized inflammation in the healthy infraorbital zone, is a response to stress (UV, pollution, fatigue, etc.).

[0005] The cutaneous microcirculation, particularly in the infraorbital zone, is organized into two parallel plexuses located less than 1 mm below the skin surface. The upper plexus, located in the papillary dermis, consists of small arterioles and venules, with capillary loops extending perpendicular to the skin surface. Capillaries are the site of oxygen and nutrient exchange in the skin tissue, while small venules play a role in leukocyte extravasation. The lower plexus is located at the dermal-subcutaneous tissue boundary. It consists of arteries and veins originating from adipose tissue and underlying muscle, which penetrate the fascia to form ascending arterioles and descending venules, which connect to the superficial plexus.

[0006] Microvascular endothelial cells are the major component of dermal blood vessels. Endothelial cells are interconnected by intercellular junctions, forming a barrier between the blood and the surrounding dermal tissue. The endothelial barrier is a dynamic structure that controls the exchange of fluids and solutes, including plasma proteins and cells, particularly leukocytes. Under basic homeostatic conditions, the intercellular junctions are poorly permeable to fluids and solutes. Furthermore, multiple regulatory processes operate in endothelial cells to maintain endothelial barrier function.

[0007] Under inflammatory conditions caused by various factors (UV, air pollution, fatigue, stress), various pro-inflammatory mediators, including TNF-α, act on endothelial cells to increase vascular permeability and open intercellular junctions, ultimately leading to extravasation of leukocytes and the leakage of red blood cells and other nutrients that make up plasma. Therefore, skin microcirculation deteriorates, and the accumulation of white blood cells in the skin tissue causes swelling under the eyes. At the same time, the accumulation of red blood cells in the extracellular space leads to purple pigmentation of the skin around the eyes, characteristic of peripheral darkening.

[0008] Hemoglobin is the main component of red blood cells and is decomposed into pigmented degradation products, such as heme (red pigment), which are released and accumulate in the dermis and epidermis. Nevertheless, the decomposition of free heme also leads to the release and accumulation of iron molecules in skin tissue. Free ferrous ions oxidize and produce ROS (reactive oxygen species), which leads to increased skin oxidation and inflammation. For this reason, chelation of ferrous ions constitutes a complementary strategy for not only decomposing heme and combating the formation of peripheral dark spots, but also improving skin radiance.

[0009] The Applicant is interested in sea buckthorn in the context of research into skin microcirculation.

[0010] Described by Linnaeus in 1753, Hippophae rhamnoides L. (APG IV 2016 Plant Classification) is a spiny, dioecious shrub native to the temperate zones of Europe and Asia. The species was introduced to Canada by Russian immigrants in the early 20th century. Recommended for preventing soil erosion, the species is now distributed throughout the territory and is cultivated for its fruit in Saskatchewan, British Columbia, and Quebec.

[0011] Hippophae rhamnoides, a shrub restricted to river banks or dune massifs, as well as the edges of sandy forests, can reach a height of 1 to 5 m and has slender, pointed, deciduous leaves with silvery undersides. The flowers are petalous, very small, and greenish, and the leaves grow after they bloom. The female stems produce fruit in the form of fleshy berries 6 to 8 mm in diameter, which cover the branches in dense clusters. The berries ripen in early autumn and have a beautiful orange color and a strong, fragrant flavor. The mature fruits remain on the twigs throughout the winter.

[0012] The morphological characteristics of sea buckthorn vary greatly in response to the wide range of climatic conditions across the taxon's distribution range. Varietal selection (ease of harvesting, high oil content, disease resistance, etc.), numerous crosses between subspecies, and genetic modification programs mean that it is no longer identifiable as a subspecies. Only horticultural cultivars, such as Leikora or Orange Energy, have defined names.

[0013] Hippophae rhamnoides, which is very widespread on the continents of the Northern Hemisphere, has several common names that refer to its morphology or its host environment: argasse, grisset, glossy thorn, purging thorn, thorny willow, false buckthorn, seaberry, olive, or Siberian pineapple. In other languages, these are sea buckthorn (English), Sanddorn (German), or espino amarillo (Spanish).

[0014] Due to limitations caused by heavy snowfall in winter, fruit harvesting in North America occurs in the fall before the leaves fall. The fruiting branches are cut, frozen, and then mechanically sorted to separate the fruit from the branches and leaves. The resulting leaf by-product is dried and used as a tea substitute.

[0015] The food, medicinal, horticultural and environmental uses of Hippophae rhamnoides date back to ancient times: the Greeks used the leaves and young twigs in supplementary feed to promote weight gain in horses and to give their coats a lustrous sheen, hence its Latin name, hippophae, which comes from "hippos" = horse and "phaos" = sheen.

[0016] In traditional Chinese, Japanese, or Tibetan medicine, as well as in Ayurvedic medicine (India), sea buckthorn is still widely used to treat all kinds of skin and mucous membrane conditions, especially as a tonic. It is also used for digestive disorders, lung inflammation, and menstrual pain or irregularities. In Tibet, in particular, the seeds, fruits, or leaves can be used to treat oral diseases, regenerate tissue, and treat inflammation and bacterial infections, while in Turkey, the fruits and leaves are used as antiseptics for healing and treating ulcers.

[0017] It is also used in combination with other plants to treat certain cardiovascular disorders (especially platelet aggregation disorders), digestive disorders, indigestion, lung inflammation, and menstrual irregularities or pain. Virtually all parts of the sea buckthorn are used in traditional medicine; in addition to the berries and seeds, leaf and bark extracts are also prepared.

[0018] Sea buckthorn leaf extract is used to prepare teas, tinctures, and other decoctions in the following situations: - Prevention and treatment of colds, tonsillitis, bronchitis and ARVI (strengthens immunity and fights seasonal diseases); - Prevention and treatment of hypertension, has sedative and hypotensive effects, is a blood pressure stabilizer; - Treatment of cardiovascular diseases; - Treatment of diabetes (hypoglycemic effect); - Treatment of inflammatory diseases of the joints; - Treatment of liver diseases (hepatoprotective effect, restoration of normal liver cells in inflammatory diseases); - Treatment of colitis, gastric ulcers and duodenal ulcers by using an infusion or powder of dried leaves.

[0019] The document FR2943255 describes an extract of sea buckthorn seeds (Hippophae rhamnoides), rich in fatty acids and sterols obtained by supercritical CO2 extraction, for stimulating 5-alpha reductase activity and sebum production to treat menopause-related cutaneous hormonal imbalances, such as loss of skin radiance. Biological activity has been demonstrated in terms of stimulation of 5-alpha reductase activity by normal human fibroblasts (NHFs), collagen I / NHF synthesis, glycosaminoglycan and hyaluronic acid / NHF synthesis, and integrin synthesis by normal human keratinocytes.

[0020] The document FR2971940 describes the aqueous, alcoholic, or glycolic extracts of winter twigs of Hippophae rhamnoides obtained for their depigmenting effect on skin, body hair, and scalp hair. "Winter twigs" simply refers to the leafless stem part of sea buckthorn, which contains indole compounds. The biological mechanism responsible for the depigmenting effect is based on the stimulation of melanin biosynthesis produced by melanocytes.

[0021] Prior art describes extracts of sea buckthorn leaves, the chemical composition of which varies qualitatively and quantitatively depending on the extraction process / operational parameters used. For example, in 2012 and 2014, Jayashankar described an extraction method using supercritical CO2 and ethanol as a cosolvent at 200 bar pressure and 50°C, producing an extract containing isorhamnetin and exhibiting anti-inflammatory effects on several targets, including IL-6. A 2017 study by Enkhtaivan et al. described a methanol extraction method that produced an extract containing aglycone-type flavonoids, such as isorhamnetin. A 2018 study by Tanwar et al. described extracts from sea buckthorn leaves using anhydrous solvents, such as 70% ethanol, which has a change in polarity compared to 96°C ethanol. This change in polarity leads to a change in the solvent's extractive power, ultimately resulting in sea buckthorn extracts with specific phytochemical compositions. A study by Sadowska et al. in 2020 described fractionated extracts obtained from sea buckthorn leaves, stems and fruits using a complex method involving different stages of evaporation and partial solubilization of the concentrate using 80% methanol, and the resulting extracts had specific qualitative and quantitative phytochemical compositions.

[0022] Documents KR20140148141 and KR101121590 describe the ethanol extraction of sea buckthorn leaves under specific operating conditions, particularly with regard to temperature and extraction time. The phytochemical composition therefore varies qualitatively and quantitatively depending on the operating parameters applied. For example, in document KR101121590, the extraction is carried out at room temperature with C1-C4 alcohols, allowing the extraction of isorhamnetin-3-O-glucoside-7-O-rhamnoside, which is responsible for the therapeutic effects, e.g., cancer prevention. [Prior art documents] [Patent documents]

[0023] [Patent Document 1] FR2943255 [Patent Document 2] FR2971940 [Patent Document 3] KR20140148141 [Patent Document 4] KR101121590 Summary of the Invention [Problem to be solved by the invention]

[0024] The present application demonstrates, quite surprisingly, that an extract of the stem-free leaves of Hippophae rhamnoides has an effect on the microcirculation of the skin and can be used to reduce the darkness and / or puffiness around the eye contour and / or to improve the radiance of the complexion. [Means for solving the problem]

[0025] More precisely, according to a first aspect, the object of the invention is the cosmetic and / or dermatological use of an anhydrous extract of Hippophae rhamnoides leaves or a cosmetic composition comprising said extract, for maintaining and / or improving cutaneous microcirculation, ultimately for reducing the darkness and / or puffiness around the eye contour and / or for maintaining and / or increasing the radiance of the skin complexion, with the aim of healthy skin.

[0026] As will be seen below, the present application demonstrates that the extract according to the invention makes it possible to: - reducing the expression of the VCAM-1 adhesion protein and / or - reducing the adhesion of monocytes to the endothelial cell membrane; and - increasing transendothelial electrical resistance, and / or - increasing the expression of the enzyme HMOX-1, and / or - increasing chelation of ferrous ions, and / or - increasing the expression of the antioxidant enzymes GPX2 and / or GPX3 and / or TXN, and / or - Increasing radical scavenging activity.

[0027] The VCAM-1 adhesion protein is responsible for the extravasation of leukocytes and erythrocytes into the extracellular space by migration between two endothelial cells, which is facilitated by disruption of the endothelial contacts that form the intercellular spaces through which the cells pass.

[0028] More precisely, leukocytes circulate in the bloodstream but must cross the endothelial barrier to reach inflamed tissue. This rapid migration of blood to the site of infection is essential for tissue repair in response to acute inflammation. Leukocyte extravasation is a highly regulated process involving complex interactions between leukocytes and the endothelium, particularly via selectins, integrins, intercellular adhesion molecule (ICAM1), vascular adhesion molecule (VCAM1), junctional adhesion molecule (JAM-1 / A / C), and platelet-endothelial cell adhesion molecule (PECAM1).

[0029] The first step in the inflammatory response is the reorganization of the endothelial cell surface to capture circulating leukocytes. The release of inflammatory cytokines stimulates the synthesis of adhesion molecules (P-selectin, E-selectin) on the endothelial cell surface, which locally promotes weak and transient adhesive interactions between leukocytes and the endothelium. The deposition of chemokines on the endothelial surface then triggers the activation of leukocyte integrin (ICAM-β2), which leads to firm adhesion of leukocytes and their arrest through interaction with surface receptors (ICAM1, VCAM1).

[0030] In other words, reducing the expression of the VCAM-1 adhesion protein can reduce the inflammatory response.

[0031] Transendothelial electrical resistance then makes it possible to measure the membrane permeability of the endothelial cells and therefore their barrier function: the higher the resistance, the more effective the barrier function.

[0032] In other words, an increase in transendothelial electrical resistance is synonymous with an enhancement of endothelial barrier function.

[0033] Regarding the HMOX-1 gene, it encodes an enzyme that breaks down hemoglobin, the main component of red blood cells and the source of the dark pigmentation that surrounds them.

[0034] As already mentioned, the accumulation of red blood cells in the extracellular space leads to purple pigmentation of the skin around the eyes, characteristic of peripheral darkening.

[0035] Hemoglobin is the main component of red blood cells, which breaks down and releases pigmented breakdown products, such as heme, that accumulate in the dermis and epidermis. Free heme is toxic when uncomplexed. Consequently, it is essential to eliminate free heme. The enzyme heme oxygenase type 1 (HMOX-1) catabolizes heme by breaking it down into biliverdin, which is then converted to bilirubin by the action of biliverdin reductase A. These catabolic products are known for their antioxidant role and help reduce the appearance of peripheral dark spots.

[0036] In other words, increasing the expression of the HMOX-1 gene makes it possible to increase the degradation of heme.

[0037] Finally, increasing the transcriptome expression of GPX2 and / or GPX3 and / or TXN oxidative defense genes makes it possible to reduce the oxidation of hemoglobin to ROS (reactive oxygen species), particularly the oxidation of ferrous ions, which accumulate in the extracellular space due to hemoglobin degradation.

[0038] According to the present invention, the extract comprises flavonoids, gall derivatives and triterpenes.

[0039] especially, - the flavonoids include glycosylated flavonols, advantageously including isorhamnetin-3-O-glucoside and narcissine; - gall derivatives include gallic acid and ellagic acid; - Triterpenes include ursolic acid and maslinic acid.

[0040] According to the present invention, - The concentration of glycosylated flavonols in the extract ranges from 20 mg / kg to 5 g / 100 g; - The concentration of gall derivatives in the extract is between 2 mg / kg and 1 g / 100 g; - The concentration of triterpenes in the extract ranges from 80 mg / kg to 7 g / 100 g.

[0041] In another aspect, the present invention relates to an anhydrous extract of stem-free leaves of Hippophae rhamnoides, comprising flavonoids, gall derivatives and triterpenes.

[0042] especially - the flavonoids include glycosylated flavonols, advantageously including isorhamnetin-3-O-glucoside and narcissine; - gall derivatives include gallic acid and ellagic acid; - Triterpenes include ursolic acid and maslinic acid.

[0043] According to the present invention, - The concentration of glycosylated flavonols in the extract ranges from 20 mg / kg to 5 g / 100 g; - The concentration of gall derivatives in the extract is between 2 mg / kg and 1 g / 100 g; - The concentration of triterpenes in the extract ranges from 80 mg / kg to 7 g / 100 g.

[0044] According to another embodiment, the extract of the invention does not contain isorhamnetin and / or isorhamnetin-7-O-glucoside and / or isorhamnetin-3-O-glucoside-7-O-rhamnoside.

[0045] In practice, the extract is obtained by a first solid extraction / solvent extraction step, followed by a second solid separation / solvent extraction step, then a third step for recovering the extract in liquid or pasty form in the presence of an anhydrous solvent advantageously chosen from the group comprising 96° ethanol or a mixture of ethanol and supercritical CO2.

[0046] The present invention also relates to an extract obtainable by the above process.

[0047] The present invention also relates to cosmetic and / or dermatological compositions comprising said extract.

[0048] In particular, solid extraction / solvent extraction may be carried out using different techniques well known to those skilled in the art, such as maceration, re-maceration, digestion, dynamic maceration, decoction, fluidized bed extraction, microwave assisted extraction, ultrasound assisted extraction, countercurrent extraction, percolation, re-percolation, percolation, vacuum extraction, shaking, supercritical fluid extraction, subcritical water extraction, reflux extraction.

[0049] In another aspect, the solid / solvent extraction is carried out from stem-free leaves in fresh, fresh-frozen or dried form, and the leaves may be in whole, crushed, ground or freeze-ground form.

[0050] Advantageously, the leaves are collected during the summer / autumn fruit harvest period.

[0051] According to the present invention, the extraction solvent is a non-polar or anhydrous solvent of intermediate polarity (i.e., containing less than 5% water). Thus, according to the present invention, the extraction solvent may be selected from the intermediate polarity group including alcohols, e.g., ethanol, glycols, e.g., propylene glycol, 1,3-propanediol and butylene glycol, glycerin, ethyl acetate, low transition temperature mixtures (LTTM), or anhydrous natural deep eutectic solvents (NaDES). Also, a range of non-polar solvents, e.g., supercritical CO2, vegetable oils, C8-C8 10They may be selected from medium chain triglycerides, fatty acid esters such as octyldodecyl myristate or 2-methyltetrahydrofuran. These solvents may be used alone or as a mixture.

[0052] In an advantageous embodiment, ethanol, advantageously at 96°, or supercritical CO2, or a mixture of the two, is used as the extraction solvent.

[0053] In an advantageous embodiment, the extraction solvent is 96° ethanol or a mixture of ethanol and supercritical CO2.

[0054] When the extraction solvent is a mixture of ethanol and supercritical CO2, the ethanol / supercritical CO2 mass ratio is advantageously comprised between 1:5 and 1:50, advantageously between 1 / 10 and 1 / 20.

[0055] In practice, the plant / solvent ratio applied in the extraction process is comprised between 1 / 99 and 80 / 20, advantageously between 2 / 98 and 20 / 80.

[0056] When the extraction solvent is a mixture of ethanol and supercritical CO2, the plant / ethanol mass ratio is advantageously comprised between 10 / 90 and 50 / 50, the plant / supercritical CO2 mass ratio is advantageously comprised between 0.5 / 99.5 and 15 / 85, and the plant / supercritical CO2-ethanol mass ratio is advantageously comprised between 2 / 98 and 10 / 90.

[0057] When the extraction solvent is 96° ethanol, the plant / 96° ethanol mass ratio is advantageously between 1 / 99 and 20 / 80.

[0058] When the extraction solvent is a mixture of ethanol and supercritical CO2, the extraction is carried out at a temperature comprised between 40°C and 60°C, preferably between 45°C and 55°C, at an absolute pressure comprised between 220 bar and 350 bar, preferably between 270 bar and 290 bar, for a time comprised between 1 hour and 5 hours, preferably between 2 hours and 4 hours.

[0059] When the extraction solvent is 96° ethanol, the extraction is carried out at a temperature comprised between 60°C and 90°C, preferably between 70°C and 85°C, at atmospheric pressure, for a time comprised between 1 hour and 5 hours, preferably between 2 hours and 4 hours.

[0060] According to the present invention, a solid extraction / solvent extraction step is carried out, followed by a solid separation / solvent extraction step, and then a step for recovering the liquid or paste phase containing the active material. This separation may be carried out using any technique known to those skilled in the art, in particular draining, pressing, dewatering, decanting (by gravity or centrifugation) or filtration.

[0061] According to another embodiment, a step for recovering the liquid or pasty phase is carried out, followed by a concentration step, which makes it possible to obtain a concentrated liquid to a pasty form according to a concentration factor, in practice carried out by evaporation at atmospheric or reduced pressure or by membrane separation.

[0062] After or simultaneously with the concentration step, the extract could be dissolved in a recovery solvent selected from the group including glycol, glycerin, ethanol, anhydrous LTTM, anhydrous NaDES, medium chain triglycerides, fatty acid esters and vegetable oils.

[0063] Advantageously, - when the extraction solvent is a mixture of ethanol and supercritical CO2, the recovery solvent is selected from the group consisting of octyldodecyl myristate, capric triglyceride and caprylic triglyceride, vegetable oils and mixtures thereof; preferably octyldodecyl myristate; When the extraction solvent is 96° ethanol, the recovery solvent is advantageously chosen from the group constituted by 1,3-propanediol, propylene glycol, butylene glycol, anhydrous LTTM and mixtures thereof; 1,3-propanediol.

[0064] In embodiments where the extraction solvent is a mixture of supercritical CO2 and ethanol, the ethanol is evaporated and the concentrated plant extract is dissolved in a recovery solvent, which is advantageously octyldodecyl myristate, advantageously in a proportion of 0.1 to 5% by weight of the dried plant extract, preferably between 0.3 and 1%, and indeed as much as 0.5%.

[0065] In embodiments where the extraction solvent is 96° ethanol, the 96° ethanol is evaporated and the plant extract is dissolved in a recovery solvent, which is advantageously a glycol, preferably 1,3-propanediol, advantageously in a proportion of the order of 0.5 to 5% by weight of the dried plant extract, preferably between 1 and 3%, preferably 1.5% by weight.

[0066] Finally, a step may be carried out to dissolve the extract for sterile or non-sterile packaging, followed by one or more filtration steps. Prior to the final filtration step, additives such as preservatives and antioxidants known to those skilled in the art may be incorporated into the liquid extract to ensure its stability.

[0067] Depending on the nature of the solvent used, the extract obtained may prove to be too dark in color, in which case the process for obtaining the extract comprises an additional step of decolorizing the extract, preferably by adsorption, advantageously on activated carbon or a decolorizing earth.

[0068] The extract is therefore intended for use in the cosmetic and / or dermatological field, advantageously in cosmetics, and is presented in a form suitable for topical administration.

[0069] The cosmetic and / or dermatological extract or the composition in which it is incorporated is therefore in a form intended for use in the cosmetic and / or dermatological field by the topical route, for reducing the darkness and / or puffiness around the eye contour and / or for maintaining and / or increasing the radiance of the skin complexion by maintaining and / or improving the cutaneous microcirculation of healthy skin.

[0070] The extract, which can be advantageously obtained directly by one of the methods described above, is suitable for use in the cosmetic field, in particular in the form of a composition used in the cosmetic and / or dermatological field.

[0071] In practice, the extract represents between 0.1% and 10% by weight of the cosmetic and / or dermatological composition, preferably between 0.5% and 5% by weight.

[0072] The cosmetic and / or dermatological compositions according to the invention may be in any of the galenical forms usually used for topical application to the skin, for example in anhydrous form, in the form of an oil-in-water emulsion, a water-in-oil emulsion, a multiple emulsion, a silicone emulsion, a microemulsion, a nanoemulsion, a gel, an aqueous solution or a hydroalcoholic solution.

[0073] The composition may be more or less fluid and may be in the form of a white or colored cream, ointment, emulsion, lotion, serum, or gel.

[0074] The cosmetic and / or dermatological compositions may contain excipients commonly used in the cosmetic and / or dermatological fields, such as fats, cleansing and / or conditioning surfactants, emulsifying and co-emulsifying agents, hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, exfoliants, fragrances, fillers, hydrophilic and lipophilic sunscreens, colorants, neutralizing agents, penetration enhancers, and polymers. These types of excipients are well known to those skilled in the art.

[0075] In practice, the amounts of these various excipients are those customarily used in the field under consideration, the sum of the excipients preferably being from 0.01% to 30% of the total mass of the composition.

[0076] Suitable fats that may be mentioned are mineral oils, oils of animal origin (e.g., lanolin), vegetable oils, synthetic oils (e.g., isopropyl myristate, octyldodecyl, isostearyl isostearate, decyl oleate, isopropyl palmitate) and silicone oils (cyclomethicone, dimethicone).Fats that may be used are fatty alcohols, fatty acids, waxes and gums, especially silicone elastomers.

[0077] Examples of suitable cleansing and / or conditioning surfactants that may be mentioned are nonionic, anionic, cationic or amphoteric surfactants, and mixtures thereof, such as alkyl sulfates, alkyl ether sulfates, for example sodium lauryl ether sulfate, alkyl betaines, for example cocamidopropyl betaine, or quaternary ammonium salts.

[0078] Examples of suitable emulsifiers and co-emulsifiers that may be mentioned are esters of polyglycerol and fatty acids, esters of sucrose and fatty acids, esters of sorbitan and fatty acids, esters of oxyethylenated fatty acids and sorbitan, ethers of fatty alcohols and PEG, esters of glycerol and fatty acids, alkyl sulfates, alkyl ether sulfates, alkyl phosphates, alkyl polyglucosides, alkyl polypentosides, dimethicone copolyols.

[0079] Examples of suitable hydrophilic gelling agents that may be mentioned are carboxyvinyl polymers, acrylic copolymers (carbomers), such as acrylate / alkyl acrylate copolymers, polyacrylamides, polysaccharides, such as xanthan gum, guar gum, natural gums, such as cellulose gum and derivatives, starch and its derivatives, clays and 2-acrylamido-2-methylpropanoic acid copolymers.

[0080] Examples of suitable lipophilic gelling agents that may be mentioned are modified clays, such as bentones, fatty acid metal salts, hydrophobic silica, and ethylcellulose.

[0081] Examples of suitable preservatives that may be mentioned are benzoic acid, sorbic acid, propionic acid, salicylic acid, dehydroacetic acid, and their salts, benzyl alcohol, ethylhexylglycerin, parabens, their salts and esters, triclosan, imidazolidinyl urea, 5-phenoxyethanol, DMDM hydantoin, diazolidinyl urea, and chlorphenesin.

[0082] Examples of suitable antioxidants that may be mentioned are chelating agents such as EDTA and its salts, sodium metabisulfite, salicylic acid, ascorbic acid, and citric acid and their salts, sodium tartrate, sodium gluconate, carotenoids, and tocopherols.

[0083] Examples of solvents (different from the extraction solvent) that may be used in the cosmetic composition that may be mentioned are water, ethanol, glycerin, propylene glycol, propanediol, butylene glycol, and sorbitol.

[0084] Examples of suitable exfoliants that may be mentioned are chemical exfoliants, such as AHAs, and physical exfoliants, such as natural or synthetic powders.

[0085] Examples of suitable fillers that may be mentioned are talc, kaolin, mica, sericite, magnesium carbonate, aluminum silicate, magnesium silicate, and organic powders such as nylon.

[0086] Examples of suitable colorants that may be mentioned are lipophilic colorants, hydrophilic colorants, pigments and pearls commonly used in cosmetic or dermatological compositions, and mixtures thereof.

[0087] Examples of suitable neutralizing agents that may be mentioned are sodium hydroxide, triethanolamine, aminomethylpropanol, and potassium hydroxide.

[0088] Examples of suitable penetration enhancers that may be mentioned are alcohols and glycols (ethanol, propylene glycol), ethoxydiglycol, alcohols and fatty acids (oleic acid), fatty acid esters, and dimethyl isosorbide.

[0089] The compositions of the invention may also comprise active ingredients other than the extract according to the invention. Examples of suitable active ingredients that may be mentioned are free radical scavengers, or more generally antioxidants, whitening agents, pigments, emollients, moisturizers, antiseborrheic agents, anti-inflammatory agents, anti-acne agents, keratolytic and / or desquamating agents, anti-wrinkle and toner agents, exfoliants, anti-irritants, soothing agents, vitamins and mixtures thereof, mattifying agents, anti-ageing active ingredients such as retinol, healing agents, disinfectants, and essential oils.

[0090] The invention and the advantages achieved will become more apparent from the following embodiments with reference to the accompanying drawings. [Brief explanation of the drawings]

[0091] [Figure 1] This is a photograph showing the colorimetric evaluation of chelation of ferrous ions (Fe2+) in solutions containing Extract 1C at concentrations of 0.1% to 2% relative to the NT solution. DETAILED DESCRIPTION OF THE INVENTION [Example]

[0092] Preparation and Phytochemical Analysis of Supercritical CO2 and Ethanol Co-Solvent Extracts (Extracts 1, 1A, 1B, and 1C) Dried leaves of Hippophae rhamnoides were crushed. The crushed and dried leaves and 96°C ethanol were introduced into a supercritical CO2 extractor in a 1:2 mass ratio. Solid / solvent extraction was performed by continuously diffusing supercritical CO2 into the extractor at a temperature of 50°C and a pressure of 285 bar, with a total supercritical CO2 charge of 30 kg CO2 / kg plant, using either a 1:15 ethanol / supercritical CO2 ratio or a 3:97 plant / solvent (supercritical CO2 and ethanol) ratio. At the extractor outlet, the fluid expanded, releasing CO2 in gaseous form, and the concentrated ethanol extract was collected in liquid form. The ethanol was then evaporated at 70°C under a reduced pressure of 50 mbar to obtain a pure, concentrated plant extract (designated Extract 1). The resulting Extract 1 was a greenish-brown paste.

[0093] This concentrated extract 1 could then be dissolved in different solvents as needed.

[0094] Extract 1A: To make this concentrated lipophilic extract bioavailable in aqueous media for biological efficacy testing, Extract 1 was completely dissolved in 5 g / 100 mL of DMSO and stirred at 60° C. for 15 minutes (Extract 1A). The resulting Extract 1A was a greenish-brown liquid.

[0095] Extract 1B: To obtain an oily liquid that can be easily formulated into cosmetics, Extract 1 was completely dissolved in octyldodecyl myristate (MOD) at a concentration of 0.5g / 100g for 1 hour at 60°C with mechanical stirring, and then filtered through a cellulose plate with a cutoff threshold between 1.5 and 3μm (Extract 1B). The resulting Extract 1B was a clear green liquid.

[0096] Extract 1C: Extract 1B may be further decolorized using activated carbon to render the oily liquid extract colorless or nearly colorless, containing only green chlorophyll pigments. 0.8% powdered activated carbon was added to Extract 1B at 50°C for 1 hour with mechanical stirring. The mixture was then filtered through a cellulose plate with a cutoff threshold between 2.5 and 4.2 μm (Extract 1C). The resulting Extract 1C was in the form of a transparent yellow liquid.

[0097] Triterpenoids were studied and quantified for all extracts by HPLC-CAD (CAD: charged aerosol detector). The presence of three important triterpenoid acids was detected: ursolic acid, oleanolic acid, and maslinic acid. These three triterpenoid acids were quantified against a calibration curve obtained with ursolic acid molecular standard. The concentration of these three triterpenoid acids in extract 1 was 5%.

[0098] All results are reported in Table 1.

[0099] [Table 1]

[0100] The study and quantification of phenolic compounds was carried out by UHPLC-UV on extracts 1 and 1A alone. The presence of MOD prevented the analysis in extracts 1B and 1C. The presence of flavonoids, e.g., glycosylated flavonols (7 molecules) and gall derivatives (3 molecules), was detected in the extracts. Their concentrations were determined against calibration curves established with narcissin molecular standards for glycosylated flavonols and ellagic acid for gall derivatives. The concentrations in extracts 1B and 1C were determined by calculation according to the dilution factor. In extract 1, the concentration was around 1%.

[0101] All the results are shown in Table 2.

[0102] [Table 2] [Example]

[0103] Preparation and phytochemical analysis of ethanol extracts (extracts 2, 2A, 2B, and 2C) Dried leaves of Hippophae rhamnoides were crushed. Crushed and dried Hippophae rhamnoides leaves and 96°C ethanol were introduced into an extractor in a 1:9 mass ratio. Solid / liquid extraction was carried out for 3 hours at atmospheric pressure and the mixture's reflux temperature with continuous mechanical stirring. Upon completion of the extraction, solid / liquid separation was carried out by filtration through a filter cloth. The crude liquid extract was then filtered through a cellulose plate to a 0.8 to 0.5 μm grade, and chlorophyll was removed by adding 0.2% activated carbon at room temperature with mechanical stirring for 1 hour. The ethanol was then evaporated at 80°C under a 50 mbar reduced pressure to obtain a concentrated, pure plant extract (designated Extract 2). The resulting Extract 2 was a brown paste.

[0104] This concentrated extract 2 could then be dissolved in different solvents as needed.

[0105] Extracts 2A and 2B: To enable the bioavailability of this concentrated extract with intermediate polarity in aqueous media for biological efficacy testing, Extract 2 was completely dissolved in DMSO at two concentrations: 10 g / 100 mL (Extract 2A) and 1.4 g / 100 mL (Extract 2B) with stirring for 15 minutes at 60° C. The extracts thus obtained were in the form of a brown liquid (Extract 2A) and a clear amber to yellow liquid (Extract 2B).

[0106] Extract 2C: To obtain an anhydrous liquid form that can be easily used by cosmetic formulators, Extract 2 was completely dissolved in 1,3-propanediol (PDO) at a concentration of 1.4 g / 100 g with mechanical stirring at 60° C. for 1 hour, and then filtered through cellulose plates to a final cut-off threshold comprised between 0.3 and 0.1 μm (Extract 2C). The resulting Extract 2C was a clear, amber to yellow liquid.

[0107] The phytochemical characterization of the extracts is shown in Tables 3 and 4.

[0108] The study and quantification of phenolic compounds in the extract was carried out by UHPLC-UV. Similar to extract 1, the presence of glycosylated flavonol-type flavonoids (7 molecules) as well as gall derivatives (3 molecules) was detected in extract 2. Their concentrations were determined against calibration curves established with narcissine molecular standard for glycosylated flavonols and ellagic acid for gall derivatives. The total concentration of identified phenolic compounds in extract 2 was approximately 4%.

[0109] All the results are shown in Table 3.

[0110] [Table 3]

[0111] The study and quantification of triterpenoids was carried out for extract 2 by HPLC-CAD (CAD: charged aerosol detector). As in extract 1, the presence of three important triterpenoid acids was also detected: ursolic acid, oleanolic acid, and maslinic acid. These three triterpenoid acids were quantified against a calibration curve obtained with a molecular standard of ursolic acid. The concentrations in extracts 2A, 2B, and 2C were determined by calculation depending on the dilution factor. The concentration of these three triterpenoid acids in extract 2 was 1%.

[0112] All results are shown in Table 4.

[0113] [Table 4] [Example]

[0114] Effect of extracts 1A and 2A according to the invention on the synthesis of the firm adhesion protein VCAM-1 in inflammatory conditions in 2D cultures of human dermal microvascular endothelial cells Principle of the method: In situ immunolabeling combined with image analysis was used to measure the synthesis of the firm adhesion protein VCAM-1 in monolayer cultures of inflamed dermal microvascular endothelial cells treated or not with Extract 1A or Extract 2A.

[0115] Protocol: Human dermal microvascular endothelial cells were pretreated for 18 hours with Extract 1A or Extract 2A at final concentrations of 0.025% and 0.07%, respectively. The cells were then stimulated with 0.2 ng / mL TNF-α and simultaneously treated with Extract 1A or Extract 2A at final concentrations of 0.025% and 0.07%, respectively, for 6 hours. Non-treated cells (NT) were used as a control. Cells were washed with phosphate-buffered saline, fixed, permeabilized with 0.1% Triton for 5 minutes, and saturated with 1% BSA (bovine serum albumin) for 1 hour. Cells were incubated overnight with primary antibody (anti-VCAM-1), washed in PBS buffer, and incubated with secondary antibody conjugated to the fluorescent dye Alexa 594 for 1 hour. Fluorescence was read using appropriate filters on a Cytation 5 imaging spectrophotometer (Biotek). Fluorescence measurements by image analysis were performed using standard acquisition parameters.

[0116] The study was based on three independent experiments (n=3). The statistical test was a Mann-Whitney non-parametric t-test to compare the synthesis of VCAM-1 in NT dermal microvascular endothelial cell cultures containing endothelial cell extracts treated with 1A or 2A.

[0117] result The results are expressed as % of NT cells and are shown in Tables 5 and 6. Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0118] [Table 5]

[0119] [Table 6]

[0120] The extracts according to the invention induced a significant decrease in the synthesis of the protein VCAM-1 in dermal microvascular endothelial cells under inflammatory conditions compared to those not treated (-82% for extract 1A vs. NT, -55% for extract 2A vs. NT). The extracting solvent alone did not induce a decrease in the synthesis of VCAM-1 (data not shown). Extracts 1A and 2A reduced vascular permeability under inflammatory conditions, i.e., the leakage of leukocytes and erythrocytes into the extracellular space, and as a result, extracts 1A and 2A induced an improvement in endothelial barrier function. [Example]

[0121] Effect of Extract 1A and Extract 2A according to the invention on reducing adult mononuclear blood cell (MNC) adhesion to human dermal microvascular endothelial cell membranes under inflamed conditions in 2D monolayer cultures Principle of the method: In situ fluorescent labeling combined with image analysis was used to measure the adhesion of MNCs to the dermal microvascular endothelial cell membrane in inflamed monolayer cultures treated or not with Extract 1A or Extract 2A.

[0122] Protocol: Human dermal microvascular endothelial cells were pretreated for 18 hours with Extract 1A or Extract 2A at final concentrations of 0.025% and 0.05%, respectively. The endothelial cells were then stimulated with 0.2 ng / mL TNF-α and simultaneously treated with Extract 1A or Extract 2A at final concentrations of 0.025% and 0.05%, respectively, for 4 hours. Non-treated cells (NT) were used as a control. In parallel with the treatment, MNCs were labeled with calcein for 1 hour and then contacted with the pre-stimulated and treated endothelial cells. After 1 hour of MNC adhesion, the endothelial cell monolayer was washed with phosphate-buffered saline, and the number of adherent MNCs was determined by reading fluorescence using appropriate filters on a Cytation 5 imaging spectrophotometer (Biotek). Fluorescence measurements by image analysis were performed using standard acquisition parameters.

[0123] The study was based on three independent experiments (n=3). The statistical test was a Mann-Whitney non-parametric t-test to compare the adhesion of MNCs to NT dermal microvascular endothelial cell membranes with that to endothelial cells treated with Extract 1A or Extract 2A.

[0124] result: The results are expressed as % of NT cells and are shown in Tables 7 and 8. Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0125] [Table 7]

[0126] [Table 8]

[0127] The extract according to the invention induced a significant decrease in the adhesion of MNCs to the dermal microvascular endothelial cell membrane in monolayer cultures under inflammatory conditions compared to those without treatment (-35% vs. N for extract 1A, -43% vs. NT for extract 2A). The extracting solvent alone did not induce a decrease in the number of adherent MNCs (data not shown). Extracts 1A and 2A reduce vascular permeability under inflammatory conditions, i.e., the leakage of leukocytes and erythrocytes into the extracellular space, and as a result, they induce an improvement in endothelial barrier function. [Example]

[0128] Effect of Extract 1A and Extract 2A according to the invention on the increase in transendothelial electrical resistance (TEER) in inflammatory conditions in 2D cultures of human dermal microvascular endothelial cells Principle of the method: The TEER (Transendothelial Electrical Resistance) technique was used to measure the in vitro membrane permeability of monolayers of dermal microvascular endothelial cells in an inflamed state, treated with or without Extract 1A or Extract 2A. TEER is a quantitative technique that measures the integrity of the endothelial cell barrier using two electrodes placed in two compartments: an electric current is passed through the monolayer and the electrical resistance of the cell barrier is measured in ohms. Thus, in a non-inflamed state, when the cells are adherent, the electrical resistance of the cells is high (enhanced endothelial barrier function), whereas in an inflamed state, when the cells are dissociated, the electrical resistance of the cells is low (defective endothelial barrier function).

[0129] Protocol: Human dermal microvascular endothelial cells were pretreated with Extract 1A or Extract 2A at final concentrations of 0.01% and 0.07%, respectively, for 24 hours. The endothelial cells were then stimulated with 0.2 ng / mL TNF-α and simultaneously treated with Extract 1A or Extract 2A at final concentrations of 0.01% and 0.07%, respectively, for 24 hours. Non-treated cells (NT) were used as a control. Endothelial barrier integrity was assessed by measuring the transendothelial electrical resistance (in ohms) of the cells in real time using a calibrated system consisting of two electrodes.

[0130] The study was based on three independent experiments (n=3).The statistical test was a Mann-Whitney non-parametric t-test comparing the transendothelial electrical resistance of NT dermal microvascular endothelial cells with that of endothelial cells treated with Extract 1A or Extract 2A.

[0131] result: The results are expressed as % of NT cells and are shown in Tables 9 and 10. Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0132] [Table 9]

[0133] [Table 10]

[0134] The extracts according to the invention induced a significant increase in transendothelial electrical resistance in monolayer cultures under inflamed conditions compared to those without treatment (+111% vs. NT for extract 1A, +160% vs. NT for extract 2A). The extracting solvent alone did not induce an increase in transendothelial electrical resistance (data not shown). Extracts 1A and 2A increased endothelial barrier function under inflamed conditions, and therefore induced a decrease in vascular permeability. [Example]

[0135] Effect of extract 1A on increasing transcriptome expression of the HMOX-1 gene encoding a hemoglobin-degrading enzyme in 2D cultures of human dermal fibroblasts Principle of the method: Real-time PCR (polymerase chain reaction) technology was used to measure the expression of the HMOX-1 (heme oxygenase 1) gene in monolayer cultures of human dermal fibroblasts treated or not with Extract 1A.

[0136] Protocol: Human dermal fibroblasts from the eyelid and under-eye bags were treated with Extract 1A at a final concentration of 0.025% for 6 hours. Non-treated cells (NT) served as a control. Cells were washed with phosphate-buffered saline and then lysed for RNA extraction. RNA was assayed to verify quality, and the relative expression of the HMOX-1 gene was assessed by real-time PCR.

[0137] The study was based on three independent experiments (n=3). The statistical test was a Mann-Whitney nonparametric t-test to compare the expression of the HMOX-1 gene in cultures of NT human dermal fibroblasts with that in cultures of dermal fibroblasts treated with Extract 1A.

[0138] result: The expression of the HMOX-1 gene was quantified in human dermal fibroblasts treated or not with Extract 1A. The results are expressed as % of NT cells and are shown in Table 11. Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0139] [Table 11]

[0140] Extract 1A induced a significant increase in HMOX-1 gene expression in human skin fibroblasts compared to untreated ones (+119% vs. NT). The extractant alone did not induce an increase in HMOX-1 expression (data not shown). Thus, Extract 1A induced the degradation of hemoglobin present in red blood cells, which is the origin of the characteristic pigmentation surrounding dark spots. [Example]

[0141] Ferrous ion (Fe) in in tubo test 2+ Effect of Extract 1C on the Chelation of Principle of the method: Ferrozine was used to evaluate the chelating power of extract 1C in an in tubular test. Ferrozine reacts with ferrous ions present in the reaction medium to form an intense purple colored Ferrozine-Fe 2+ A complex is formed. Quantification of this complex by spectrophotometry at 562 nm in a medium of known iron concentration provides information on the amount of unchelated iron and therefore on the chelating power of Extract 1C. Thus, the lighter the color of the solution containing Extract 1C, the greater the chelating power of the extract tested.

[0142] Protocol: Extract 1C was contacted with a solution of iron chloride (FeCl2) according to the concentration range of 0.1% to 2% for 10 minutes. Then, ferrozine was added to the mixture for 10 minutes to evaluate the chelating power of the extract. The absorbance of the solution was read at 562 nm by a spectrophotometer. The lower the absorbance of the solution, the higher the Fe content. 2+ The chelating power of extract 1C for ions is increased.

[0143] result: The results of solutions containing 0.1% to 2% of Extract 1C compared with the NT solution are shown in Figure 1.

[0144] Extract 1C obtained according to the present invention induced chelation of ferrous ions in a dose-effect relationship. The extraction solvent alone did not induce chelation of ferrous ions (data not shown). Thus, Extract 1C reduces pigmentation and skin tone characteristic of peripheral dark spots by chelating ferrous ions that accumulate in the extracellular space due to the degradation of hemoglobin, the main component of red blood cells. [Example]

[0145] Effect of extract 2B on radical scavenging activity by measuring DPPH in tubo Principle of the method: The antioxidant capacity of extract 2B was evaluated using an in tubular colorimetric test for DPPH (reagent name: 1,1-diphenyl-2-picrylhydrazyl). DPPH is a highly stable nitrogen radical characterized by an intense purple color that decolorizes when reduced in the presence of antioxidant molecules. The reducing power of the extract was quantified by spectrophotometrically measuring the change in absorbance of the DPPH solution after reaction with extract 2B.

[0146] Protocol: Extract 2B was contacted with solutions containing DPPH at concentrations ranging from 0.1% to 2% for 30 minutes. The absorbance of the solution was read at 518 nm using a spectrophotometer. The lower the absorbance of the solution, the greater the radical scavenging activity of Extract 2B.

[0147] The study was based on three independent experiments (n=3). The statistical test was a Mann-Whitney non-parametric t-test to compare the radical scavenging activity of NT conditions containing extract 2B.

[0148] result: The results are expressed as % of the NT condition and are shown in Table 12. Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0149] [Table 12]

[0150] Extract 2B obtained according to the present invention induced an increase in radical scavenging activity in a dose-effect relationship. The extraction solvent alone did not induce an increase in radical scavenging activity (data not shown). Extract 2B has antioxidant capacity by reducing the oxidation of iron that accumulates in the extracellular space, which is responsible for the characteristic pigmentation of peripheral dark spots and skin radiance resulting from the degradation of hemoglobin, the main component of red blood cells. [Example]

[0151] Effect of extract 2C on transcriptome expression of oxidative defense genes in 2D cultures of normal human primary keratinocytes Principle of the method: This was the same as Example 5, except for the GPX2, GPX3 and TXN genes of interest, which encode glutathione peroxidase 2, glutathione peroxidase 3 and thioredoxin, respectively.

[0152] Protocol:

[0153] Normal human primary keratinocytes were treated with Extract 2C at concentrations ranging from 0.1% to 0.5% for 24 hours. Non-treated cells (NT) served as a control. Cells were washed with phosphate-buffered saline and then lysed for RNA extraction. RNA was assayed to verify quality, and the relative expression of GPX2, GPX3, and TXN genes was assessed by real-time PCR.

[0154] The study was based on three independent experiments (n=3). The statistical test was a Mann-Whitney non-parametric t-test to compare the radical scavenging activity of NT conditions containing extract 2C.

[0155] result: The results are expressed as % of NT cells and are shown in Table 13 (GPX2), Table 14 (GPX3), and Table 15 (TXN). Values are expressed as the mean ± standard deviation across three experiments (n=3).

[0156] [Table 13]

[0157] [Table 14]

[0158] [Table 15]

[0159] Extract 2C obtained according to the present invention induced a significant increase in the transcriptome expression of GPX2, GPX3, and TXN genes in a dose-effect relationship in 2D cultures of normal human primary keratinocytes. The extractant alone did not induce an increase in the expression of GPX2, GPX3, and TXN genes (data not shown). Extract 2C has antioxidant capacity by reducing the oxidation of iron that accumulates in the extracellular space, which is responsible for the characteristic pigmentation and skin tone of dark circles resulting from the degradation of hemoglobin, the main component of red blood cells.

Claims

1. Use of an anhydrous extract of stem-free leaves of Hippophae rhamnoides or a cosmetic composition comprising said extract for use in reducing the darkness and / or puffiness around the eye contour and / or maintaining and / or increasing the radiance of skin complexion.

2. 2. Use according to claim 1, characterized in that the extract comprises flavonoids, gall derivatives and triterpenes.

3. - the flavonoids include glycosylated flavonols, advantageously including isorhamnetin-3-O-glucoside and narcissine; - gall derivatives include gallic acid and ellagic acid; - Triterpenes include ursolic acid and maslinic acid 3. Use according to claim 2, characterized in that

4. - the concentration of glycosylated flavonols in the extract is between 20 mg / kg and 5 g / 100 g; - the concentration of gall derivatives in the extract is between 2 mg / kg and 1 g / 100 g; - The concentration of triterpenes in the extract ranges from 80mg / kg to 7g / 100g 4. The use according to claim 3, characterized in that

5. An anhydrous extract of stem-free leaves of Hippophae rhamnoides containing flavonoids, gall derivatives and triterpenes.

6. - the flavonoids include glycosylated flavonols, advantageously including isorhamnetin-3-O-glucoside and narcissine; - gall derivatives include gallic acid and ellagic acid; - Triterpenes include ursolic acid and maslinic acid The extract according to claim 5, characterized in that

7. - the concentration of glycosylated flavonols in the extract is between 20 mg / kg and 5 g / 100 g; - the concentration of gall derivatives in the extract is between 2 mg / kg and 1 g / 100 g; - The concentration of triterpenes in the extract ranges from 80mg / kg to 7g / 100g The extract according to claim 6, characterized in that

8. A first solid extraction / solvent extraction step, followed by a second solid separation / solvent extraction step, then 96° ethanol or ethanol and supercritical CO 2 8. The extract according to any one of claims 5 to 7, characterized in that it may be obtained by a process comprising a third step for recovering the extract in liquid or pasty form in the presence of an anhydrous solvent selected from the group comprising mixtures of

9. The extraction solvent is ethanol and supercritical CO 2 9. Extract according to claim 8, characterized in that when the extract is a mixture of , the extract is carried out at a temperature comprised between 40°C and 60°C, preferably between 45°C and 55°C, at an absolute pressure comprised between 220 bar and 350 bar, preferably between 270 bar and 290 bar, for a time comprised between 1 hour and 5 hours, preferably between 2 hours and 4 hours.

10. The extraction solvent is ethanol and supercritical CO 2 When the mixture is with the plant / ethanol mass ratio between 10 / 90 and 50 / 50, the plant / supercritical CO 2 The mass ratio is between 0.5 / 99.5 and 15 / 85, and the plant / supercritical CO 2 10. Extract according to claim 8 or 9, characterized in that the -ethanol mass ratio is advantageously comprised between 2 / 98 and 10 / 90.

11. Anhydrous extraction solvent is supercritical CO 2 and ethanol, the ethanol is evaporated and the extract is dissolved in a recovery solvent selected from the group consisting of octyldodecyl myristate, capric triglyceride and caprylic triglyceride, vegetable oils and mixtures thereof; preferably octyldodecyl myristate.

12. 2. Extract according to claim 1, characterized in that when the anhydrous extraction solvent is 96° ethanol, the extraction is continuous and is carried out at a temperature comprised between 60°C and 90°C, preferably between 70°C and 85°C, at atmospheric pressure, for a time comprised between 1 hour and 5 hours, preferably between 2 hours and 4 hours.

13. 13. Extract according to claim 8 or 12, characterized in that when the extraction solvent is 96° ethanol, the plant / 96° ethanol mass ratio is comprised between 1 / 99 and 20 / 80.

14. 14. Extract according to any one of claims 8 or 12 to 13, characterized in that when the anhydrous extraction solvent is 96° ethanol, the ethanol is evaporated and the extract is dissolved in a recovery solvent selected from the group constituted by 1,3-propanediol, propylene glycol, butylene glycol, anhydrous LTTM and mixtures thereof; advantageously 1,3-propanediol.

15. Extract according to any one of claims 8 to 14, characterized in that the process further comprises a step for decolorizing the extract, preferably by adsorption, advantageously on activated carbon or a decolorizing earth.

16. A cosmetic composition comprising an extract according to any one of claims 5 to 15, characterized in that the extract represents between 0.1% and 10% by weight of the composition, preferably between 0.5% and 5% by weight.

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