HYDRO-ALCOHOLIC EXTRACT OF SCHINUS MOLLE, COSMETIC COMPOSITIONS COMPRISING SAME AND THEIR COSMETIC U

专利摘要:
The invention relates to a hydro-alcoholic extract of the aerial parts of Schinus molle. The invention is characterized in that it comprises mainly quercitrin and miquelianin among the polyphenols extracted, in a concentration of at least 0.04% of quercitrine by weight of the total weight of the extract and at least 0.02% miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently of each other being present in the form of traces. The invention also relates to a process for the preparation of a Schinus soft extract according to the invention as well as to cosmetic compositions comprising an effective quantity of the extract according to the invention as protective active agent, and the cosmetic use of such compositions. to improve the barrier function of the skin and protect against atmospheric pollutants.
公开号:FR3034664A1
申请号:FR1553082
申请日:2015-04-09
公开日:2016-10-14
发明作者:Isabelle Imbert；Nouha Domloge；Jean-Marie Botto；Remi Laville；Sebastien Garnier
申请人:Jafer；ISP Investments LLC；
IPC主号:

专利说明:

[0001] The invention relates to an aqueous-alcoholic extract of Schinus molle, its process of preparation, cosmetic compositions comprising it and their cosmetic uses. BACKGROUND OF THE INVENTION Schinus soft, also commonly known as "false pepper" or "wild pepper" or "California pepper tree" or "pepper tree of Peru", is a tree of the family Anacardiaceae originating from the arid zone of north of South America and the Andean deserts of Peru, and can be found as far as Argentina and Chile. It has also been introduced in most tropical and subtropical regions of the world. 15 Schinus molle is a fast-growing, evergreen tree, 15 meters high and 5 to 10 meters wide. The upper branches are particularly inclined to fall. The leaves are arranged alternately on the branches. They are generally 8-25 cm long and 4-9 cm wide, are finely cut, imparipinnate in shape, composed of numerous leaflets with full outline and pinnately venation, and have a strong pepper odor when wrinkled. . It is spring-flowering with small, pendulous, white-colored flowers at the ends of the branches that give rise to 5-7 mm diameter pink berries with a peppery odor. Its crooked, gnarled trunk has a rough greyish bark that also has a peppery smell. In traditional medicine, Schinus molle has in particular been used for its antibacterial and antiseptic properties in the treatment of wounds and infections (Ferreroa et al., "Acute and subacute toxicity evaluation of ethanolic extract from fruits of Schinus molle in rats", 2007). ). It also appears to have been used as an antidepressant (Machado et al., "Antidepressant-like effect of rutin isolated from ethanolic extract from Schinus molle L. in mice: 3034664 2 Evidence for the involvement of the serotonergic and noradrenergic systems", 2008 ), diuretic, against toothache, rheumatism and menstrual disorders. The fruits, leaves and sap of Schinus soft have been used for their antihypertensive, diuretic, antispasmodic and analgesic properties (Ambasata et al., "The useful plants of India", 1986, Bello et al. in vitro pharmacological evaluation of the dichloromethanol extract from Schinus molle L. ", 1998).
[0002] The insecticidal properties of Schinus soft can be considered as an alternative to synthetic chemicals in pest control (Ferreroa et al., "Acute and subacute toxicity evaluation of ethanolic extract from fruits of Schinus soft in rats", 2007). For this purpose, the insecticidal activity of aqueous and ethanolic extracts of soft Schinus leaves was tested on the elm galley (Huerta et al., "Toxicity and repellence of aqueous and ethanolic extracts from Schinus molle on elm leaf"). beetle Xanthogaleruca luteola ", 2010). It should be noted that the aqueous extract and the ethanolic extract have strictly different effects with a repellent effect for the aqueous extract whereas the ethanolic extract has an insecticidal effect. Other publications relate to the antioxidant, antimicrobial and toxicological properties of Schinus molle essential oil (eg Martins Mdo et al., "Antimicrobial and toxicological properties of Schinus molle L. essential oils", 2013). In particular, ethanolic and hexane extracts of Schinus soft leaves have been tested for toxicity in rats to evaluate their cutaneous toxicity (Bras et al., Evaluation of the acute dermal exposure of the ethanolic and hexanic extracts from the leaves of Schinus molle var areira in rats ", 2011). A mild and reversible skin irritation has been observed.
[0003] The main compounds which have been isolated from Schinus molle are triterpene ketoacids (Pozzo-Balbi et al., "Triterpenoid acids of Schinus molle", 1978). A number of mono-, di-, sesqui- and triterpenes, flavonoids, gallotannins and fatty acids have been reported in Schinus molle (Mnsel et al., "Hagers Handbuch der pharmazeutischen praxis", 1994; Terhune et al. al, "13-spathulene: a new sesquiterpene in Schinus molle or l", 1974; Dominguez et al., "Lignoceric acid and other compounds of Schinus molle", 1971). Another study led to the isolation of 3 anti-inflammatory agents, 2 triterpenic acids and one biflavanone (Yueqin et al., "Isolation of two triterpenes and a biflavanone with an antiinflammatory activity of Schinus soft fruit", 2003). 2D-PC screening of an extract obtained by extraction in a water and 80% methanol mixture of soft Schinus leaves revealed a complex mixture of glycosylated flavonols and more particularly glycosylated derivatives of quercetin (Hiermann et al. , "Die Untersuchung Potentier Wirkstoffe in Epilobium Arten.", 1983). The publication Marzouk et al. ("Antioxidant flavonol glycosides from Schinus molle", 2006) has more particularly demonstrated from an aqueous extract 80% methanol soft Schinus leaves by chromatographic separation, 2 new glycosides quercetin acyl: isoquercitrin 6 "- Op-hydroxybenzoate and 2 "-O-alpha-L-rhamnopyranosyl-hyperin 6" -O-gallate, as well as 12 known polyphenol metabolites in this species, namely gallic acid, methyl gallate, chlorogenic acid, , 2 "-alpha-L-rhamnopyranosyl-hyperin, quercetin 3-0-betaD-neohesperidoside, miquelianine, quercetin 3-O-beta-D-galacturonopyranoside, isoquercitrin, hyperine, isoquercitrin 6 "-gallate, 6" -0-gallate hyperine and (+) - catechine. Numerous publications thus relate to various extracts of Schinus molle but none seems to disclose or even suggest cosmetic use, and more particularly to improve the barrier function of the skin and to protect against atmospheric pollutants, and thus, for example, to fight against appearance of signs of skin aging. The patent document JP04176912 is also known. It describes a composition comprising one or more extracts of 5 plants selected from Aguaje (Mauritia flexuosa L. f., Achira), (Canna edulis Ker Gawl.), Algarrobo (Prosopis pallida DC.), Eight ° (Genipa americana L. ), Oca (Oxalis tuberosa Mo 1.), 011uco (Ullucus tuberousus Caldas), Cupuazu (Theobroma grandiflorum K. Schu m.), Tarwi (Lupinus mutabilis 10 Sweet), Maca (Lepidium meyenii Walpers), Mashua (Too aolol tuberosum Ruizet Pay.), Molle (Schinus molle L.) and Yacon (Polymnia sonchi folia Poepp., And Endl.). Such compositions have an effect on water retention to prevent against, for example, skin dryness, to limit itching and to impart shine to the skin and hair. However, the solvents used for maceration at room temperature for 3-7 days are those usually used by those skilled in the art and are more particularly selected from water, ethanol, 1,3-butylene glycol, propylene glycol, glycerol, and polyethylene glycol. Considering the foregoing, a problem to be solved by the invention is to develop products based on a Schinus soft extract which can be used in cosmetics, which are easy to implement, by optimizing the physico-chemical constants exhibiting beneficial properties for the skin while limiting the undesirable effects (toxicity) observed in the prior art.
[0004] In particular, the Applicants have developed a Schinus molle extraction process which has been optimized to select and guarantee in the extract obtained a high content of target molecules of interest, and specifically 2 polyphenols of interest, one of which has never been described in Schinus molle. This development was carried out according to ecoextraction criteria (biosourced solvents, optimization of heating temperature and stirring time, ratio of amount of plant material / amount and type of solvent) to obtain the best compromise polyphenolic content / energy cost ( the solvent being advantageously preserved in the liquid extract), and in order to increase the concentration of compounds of interest relative to the other polyphenols extracted with regard to the extracts known from the prior art. Of the types of extraction cited in the prior art, namely aqueous, methanolic, ethanolic, hexane or steam (essential oil) extraction or supercritical CO2 extraction of Schinus molle, none allow the selective extraction of the phenolic compounds of interest according to the invention. Indeed, the supercritical CO2 extraction allows the extraction of the oily phase and not the polyphenols of interest. Likewise, an aqueous extraction mainly extracts polar compounds such as low molecular weight sugars, proteins and polar metabolites, whereas the polyphenols of interest are present only in trace form because of their polarity. Finally, alcoholic solvents such as methanol and ethanol non-selectively extract polar and apolar molecules present in Schinus molle such as flavonols glycosides (isoquercitrin-6 "-Op-hydroxybenzoate, 2" -0-aL- rhamnopyranosyl-hyperin-6 "-O-gallate, gallic acid, methyl gallate, chlorogenic acid, 2" -αH-rhamnopyranosyl-hyperin, quercetin-3-0-13-D-neohesperidoside, miquelianin, quercetin-3-0-13 -D-galacturonopyranoside, isoquercitrin, hyperine, isoquercitrin 6 "-gallate, hyperine 30 6" -O-gallate, (+) - catechine), essential oil, steroid triterpene, preisocalamenediol, fatty acids (linoleic, linolenic, behenic, lignoceric acids ), unsaponifiable (α-amyrin, 13-sitosterol), sugars (laccase: arabinose, xylose, mannose, galactose, glucose, glucosamine).
[0005] The invention therefore firstly relates to a hydroalcoholic extract of the aerial parts of Schinus molle characterized in that it comprises mainly quercitrin and miquelianin among the polyphenols extracted, in a concentration of at least 0.04% of quercitrin by weight of the total weight of the extract and at least 0.02% of miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently of each other being present in the form of traces. In addition, a second object of the invention is a process for obtaining a soft Schinus extract according to the invention, comprising the following steps according to which: a) the aerial parts of Schinus molle are collected; b) the aerial parts of Schinus soft collected, fresh or dried, are dispersed in a proportion of 0.1% to 20% by weight of solid matter relative to the total weight used, in a hydro-alcoholic solvent, alcohol being chosen from ethanol, glycerol or propanediol, in a proportion of between 10% and 90% by weight of alcohol relative to the total weight of the solvent; c) a solid-liquid extraction is carried out, with stirring, at a temperature between 4 and 100 ° C for a period of between 10 minutes and 4 hours; d) separating the liquid and solid phases so as to eliminate the solid phase and recovering a hydroalcoholic liquid Schinus molle extract comprising mainly quercitrin and miquelianin among the extracted polyphenols, in a concentration of at least 0, 04% quercitrin by weight of the total weight of the extract and at least 0.02% of miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently from each other being present in the form of traces; and e) optionally, when the alcohol is ethanol, the liquid Schinus soft extract obtained is dried so as to obtain a solid Schinus soft extract. The third subject of the invention is a cosmetic composition, characterized in that it comprises, as protective active agent, an effective amount of a Schinus soft extract according to the invention, and a physiologically acceptable excipient. . Finally, the fourth object of the invention is the cosmetic use of a composition according to the invention for improving the barrier function of the skin and protecting against atmospheric pollutants. The invention and the advantages which follow from it will be better understood on reading the description and the non-limiting embodiments which follow, written with reference to the appended figures in which: FIG. 1 represents the evaluation of the effect protecting a soft Schinus extract according to the invention on keratinocytes subjected to environmental stress by PM10-like polluting particles; FIG. 2 represents the evaluation of the protective effect of a Schinus soft extract according to the invention on keratinocytes subjected to environmental stress by PM2.5-like polluting particles; 3 represents the effect of a soft Schinus extract according to the invention on the regulation of the sebum secretion by measuring the surface of the spots; - Figure 4 represents the evolution of the secretion of sebum by measuring the number of spots after application of a Schinus soft extract according to the invention; Figure 5 shows the effect of a Schinus soft extract according to the invention on the evolution of the number of wrinkles; - Figure 6 shows the effect of a Schinus soft extract according to the invention on the evolution of the average depth of wrinkles; Figure 7 shows the effect of a Schinus soft extract according to the invention on the evolution of skin hydration; and - Figure 8 shows the moisturizing effect of a Schinus soft extract according to the invention.
[0006] In this description, unless otherwise specified, it will be understood that, when an interval is given, it includes the upper and lower limits of said interval. The invention relates to a hydro-alcoholic extract of the aerial parts of Schinus molle comprising mainly quercitrin and miquelianin among the polyphenols extracted, in a concentration of at least 0.04% of quercitrine by weight of the total weight of the extract and at least 0.02% of miquelianin by weight of the total weight of the extract, the other extracted polyphenols taken independently of each other being present in trace form. Of the 2 polyphenols predominantly extracted according to the invention, quercitrin, which is a known flavonol glycoside, has never been mentioned in the composition of Schinus molle, and more particularly the aerial parts.
[0007] Other polyphenols extracted include those commonly present in Schinus molle such as for example isoquercitrin. Trace means a concentration for each extracted polyphenol taken independently of less than 0.01% (100 ppm) by weight of the total weight of the extract. The aerial parts of soft Schinus refer not only to the leaves themselves, but also advantageously to the unadjusted leaves of the small stems carrying the leaves.
[0008] The Schinus soft extracts according to the invention are, however, very difficult to characterize as such since their composition varies, for the same species, according to various factors such as the parts of Schinus molle used, the place of harvest, the method of harvesting. the climatic conditions, or the year of harvesting. Advantageously, the soft Schinus preferably used according to the invention is cultivated in areas remote from urban pollution, for example at altitude in South America, the extract obtained according to US Pat. The invention does not include micropollutants such as phthalates, pesticides, polycyclic aromatic hydrocarbons or heavy metals. It is particularly harvested from June to September. The extract according to the invention is obtained from the fresh or dried aerial parts of Schinus molle by solid-liquid extraction, with stirring, of 0.1% to 20% by weight of solid matter relative to the total weight put into in a hydro-alcoholic solvent, the alcohol being chosen from ethanol, glycerol or propanediol, in a proportion of between 10% and 90% by weight of alcohol relative to the total weight of the solvent, at a concentration of a temperature of between 4 and 100 ° C. for a period of between 10 minutes and 4 hours, under vacuum, at atmospheric pressure or under pressure, advantageously at atmospheric pressure, and by separation of the liquid and solid phases so as to eliminate the solid phase and recover a liquid Schinus soft extract. By fresh is meant the phase after collection during which the plant has a moisture similar to that of the plant before picking.
[0009] By drying is meant the phase after collection during which the plant is dehydrated to a so-called backup moisture. This desiccation is intended to sufficiently reduce the water content including the aerial parts (leaves and / or stems) to ensure favorable conditions for storage or subsequent processing of the plant. By way of example, the aerial parts can be naturally dried by exposure to air (in the sun or in the shade) by controlling the relative humidity for a period of not more than 10 to 20 days or, advantageously, gently drying artificially with air blown by a dryer, hot air generator or fan, at a temperature between 20 ° C and 40 ° C. Preferably, after having been advantageously washed, the aerial parts are dried, used whole or ground, preferentially crushed or even cryobrilled.
[0010] After the collection step, optionally washing, and advantageously drying and grinding of the aerial parts of Schinus molle, a dispersion and extraction step in a hydro-alcoholic solvent is carried out.
[0011] The extraction is advantageously carried out always with stirring, thus allowing dispersion and homogenization of the solid in the liquid, effectively improving the diffusion of the solute in the solvent. The water used for the extraction is a distilled water, demineralized or a water rich in mineral salts and / or trace elements, preferably distilled water. To extract quercitrin and miquelianin of interest mainly from polyphenols extracted from Schinus molle, an alcoholic co-solvent chosen from ethanol, glycerol or propanediol, for example 1,2-propanediol or 1,3-propanediol. , preferably 1,3propanediol, in a proportion of between 10% and 90% by weight of alcohol relative to the total weight of the solvent is used.
[0012] Preferably, the hydro-alcoholic solvent is a water / alcohol mixture in a proportion of between 50% and 80% by weight of alcohol relative to the total weight of the solvent, more preferably between 60% and 80% when the alcohol is ethanol or propanediol.
[0013] The solid-liquid extraction is preferably carried out at a temperature between 10 and 80 ° C, more preferably between 20 and 70 ° C, for a period of between 30 minutes and 3 hours, more preferably between 1 hour and 2 hours.
[0014] The extract according to the invention is preferably obtained from 1% to 15%, more preferably from 5% to 10%, by weight of solid material relative to the total weight used. At the end of the extraction, the plant material, which is residual and is depleted of compounds of interest, is advantageously separated from the liquid phase, for example by decantation, by centrifugation or by clarifying filtration. The liquid phase obtained can be microfiltered using filter media of suitable porosity in order to obtain a clear solution free of plant solid particles. This liquid-solid separation step may advantageously be followed by one or more purification steps, for example by microfiltration, ultrafiltration and / or nanofiltration, making it possible to concentrate the compounds of interest at the expense of other extracted compounds such as proteins. and polysaccharides, as well as solvent. The duration and mode of extraction thus play an essential role on the extracted substances, namely mainly quercitrin and miquelianin among the extracted polyphenols, the other extracted polyphenols being present in the form of traces. The hydroalcoholic extraction for a necessary and sufficient time advantageously makes it possible to solubilize quercitrin and miquelianin in an amount effective in the extract for their anti-aging protective properties. Although the alcoholic co-solvents used according to the invention have the same solvent power, the alcoholic co-solvent preferentially used is glycerol or propanediol, more preferably propanediol, still more preferably 1,3-propanediol. According to a first advantageous embodiment of the invention, such alcoholic co-solvents (glycerol or propanediol) make it possible to obtain a liquid extract in which the co-solvent is preserved. The resulting extract is thus physiologically acceptable and directly usable for cosmetic applications. According to a second embodiment of the invention, when the alcohol is ethanol, the liquid Schinus soft extract obtained is preferably dried, for example by atomization, lyophilization or zeodration so as to obtain an extract of Schinus molle solid, the ethanol being evaporated. Drying can be carried out in the presence of a carrier such as maltodextrin.
[0015] To this end, according to another aspect, the invention relates to a process for obtaining a Schinus soft extract according to the invention, according to the steps as described above, advantageously according to the first or the second mode of production.
[0016] Preferably, the process for obtaining a Schinus soft extract according to the invention comprises the following steps according to which: a) the aerial parts of Schinus molle are collected; b) the collected aerial parts of Schinus soft 15, fresh or dried, are dispersed in a proportion of 0.1% to 20% by weight of solid matter relative to the total weight used, in a hydro-alcoholic solvent, the alcohol being selected from ethanol, glycerol or propanediol in a proportion of between 10% and 90% by weight of alcohol relative to the total weight of the solvent; c) a solid-liquid extraction is carried out, with stirring, at a temperature between 4 and 100 ° C for a period of between 10 minutes and 4 hours, under vacuum, at atmospheric pressure or under pressure, preferably at atmospheric pressure; d) separating, for example by decantation, centrifugation or clarifying filtration, the liquid and solid phases so as to eliminate the solid phase and recovering a liquid aqueous Schinus molle alcoholic extract comprising mainly quercitrin and miquelianin from the polyphenols extracted in a concentration of at least 0.04% of quercitrin by weight of the total weight of the extract and at least 0.02% of miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently each other being present in the form of traces; and e) optionally, when the alcohol is ethanol, the liquid Schinus soft extract obtained in such a manner as to obtain a solid Schinus soft extract is dried, for example by atomization, lyophilization or zeodration.
[0017] The liquid Schinus soft extract thus obtained can be purified by microfiltration, ultrafiltration and / or nanofiltration to concentrate the extract into quercitrin and miquelianin relative to the proteins and polysaccharides also extracted.
[0018] By way of illustration, preferred examples of embodiments of the invention are described below. EXAMPLE 1 Preparation of a Hydro-Glycolic Extract from the Aerial Parts of Soft Schinus The aerial parts of Schinus molle, previously dried, are cryobrilled in a knife mill having a 1 mm grid, which makes it possible to obtain a powder whose particle size is predominantly around 500 μm.
[0019] The powder (150 g) thus obtained is then placed in a glass reactor provided with stirring and heating by a double jacket using a heat transfer fluid. The solvent used (1350 g) for maceration is a mixture of 70% 1,3-propanediol (DuPont Tate & Lyle Bio Products Company) and 30% distilled water. The powder of aerial parts is dispersed in the solvent for 2 hours at 50 ° C and the mixture is filtered. The extract in a solvent medium thus obtained, ie 945 g, is then determined by the Folin-Ciocalteu method which makes it possible to assay all the polyphenols. The method uses gallic acid as standard and the results are expressed in gallic acid equivalent weight%. This extract therefore contains 0.43% w / w gallic acid equivalent of total polyphenols, and more particularly 0.1% w / w of quercitrin and 0.06% w / w of miquelianine for a solids content of 2.4%. m / m, ie 18% of total polyphenols, including 4.2% quercitrin and 2.5% miquelianin in the dry extract. EXAMPLE 2 Preparation of a Hydroethanolic Extract from the Aerial Parts of Soft Schinus The aerial parts of Schinus molle, previously dried, are cryobrilled in a knife mill having a 1 mm grid, which makes it possible to obtain a powder whose particle size is predominantly 10 to 500 μm. The powder (150 g) thus obtained is placed in a glass reactor, provided with stirring and heating by double jacket using a heat transfer fluid. The solvent used (1350 g) for maceration is 70% ethanol and 30% distilled water. The powder of aerial parts is dispersed in the solvent for 2 hours at 50 ° C and the mixture is filtered. The ethanol extract is evaporated in a rotary evaporator to remove most of the solvent and then freeze-dried to obtain a solid which will then be milled. 13 g of dry extract of aerial parts of Schinus molle, assayed by the Folin-Ciocalteu method and containing 14% (gallic acid equivalent) of total polyphenols, and more particularly 4% of quercitrin 25 and 2% of miquelianine in the dry extract. In comparison, the Applicants have carried out other extraction methods for obtaining a Schinus soft extract.
[0020] For example, an ethanol extract, obtained by extraction in pure ethanol, of aerial parts of Schinus molle made under conditions comparable to those of Example 2 above makes it possible to obtain a solids content containing only 8% of total polyphenols, and more particularly 3% quercitrin and 1.2% miquelianin in the dry extract.
[0021] The process according to the invention, and in particular the choice of the hydro-alcoholic solvent, makes it possible to limit substantially or even to avoid extracting the fatty fraction extracted from soft Schinus leaves with the only alcoholic solvent (100%) and thus to concentrate the extracted polyphenols. From these results, it can be noted that the extraction mode, which is easy to implement, is essential for obtaining a soft Schinus extract comprising mainly quercitrin and miquelianin among the extracted polyphenols, in one embodiment. concentration of at least 0.04% of quercitrin by weight of the total weight of the extract and at least 0.02% of miquelianine, the other extracted polyphenols taken independently of each other being present in the form of traces, and thus be used in cosmetics. The polyphenols predominantly extracted contained in the Schinus soft extract according to the invention are more particularly anti-aging active agents which act in particular to improve the barrier function of the skin and to protect against atmospheric pollutants. Indeed, the tissues in contact with the external environment such as the skin, the hair, the nails and the lungs are directly and constantly exposed to pollutants present in the environment and potentially harmful.
[0022] In the context of the invention, atmospheric pollutants denote pollutants present in the environment, which are harmful to health and are in particular in the form of gases and respirable particles. They may be present both externally, for example diesel engine particles, ozone, or heavy metals, and / or inside houses where pollution may be due to cigarette or solvent released by paints, glues or wallpapers, such as toluene, styrene, xylene, benzaldehyde.
[0023] Among the pollutants present in the environment, there are the primary pollutants that are directly derived from pollution sources (road traffic, industries, heating, agriculture, etc.) and the secondary pollutants that come from chemical reactions of gas between them. Among the primary pollutants, mention may be made, for example, of carbon oxides, sulfur oxides, nitrogen oxides, light hydrocarbons, volatile organic compounds (VOCs), metals (lead, mercury, cadmium, etc.). ), and fine or ultrafine particles, such as PM10 (less than 10 μm in diameter) and PM2.5 (less than 2.5 μm in diameter) which can be particularly toxic because of their ability to penetrate deeply into the body. Respiratory, cardiovascular and cutaneous systems, their size being smaller than the pore size of human skin. Secondary pollutants include secondary particles, ozone and oxides of nitrogen.
[0024] The extraction according to the invention makes it possible to obtain an extract enriched in anti-aging active compounds, limpid and free from plant solid particles, advantageously stable in terms of appearance, color and odor, which can be used in the formulation of cosmetic compositions.
[0025] Advantageously, according to a third aspect of the invention, the Schinus soft extract is used in the composition of a cosmetic product comprising, as a protective active agent, an effective amount of a soft Schinus extract according to US Pat. invention, and a physiologically acceptable excipient. The term "effective amount" refers to the minimum amount of extract according to the invention which is necessary to obtain the activity especially of barrier function and protection of the skin against atmospheric pollutants, without this amount being toxic. Advantageously, the Schinus soft extract is present in the composition at a concentration of 0.001 to 1% by weight relative to the total weight of the composition, preferably from 0.01 to 1%.
[0026] Even more preferably, the composition according to the invention comprises mainly quercitrin and miquelianin, and does not substantially comprise other polyphenols or only in the form of traces, among the polyphenols extracted. An external carrier vehicle of the present physiologically acceptable step means a contact with the mucosal layers, i.e., which does not and does not cause irritation, skin or toxicity. still intolerance reaction. A physiologically acceptable excipient may comprise one or more compounds. The composition that can be used according to the invention can be applied by any appropriate route, in particular oral or external topical, and the formulation of the compositions will be adapted by those skilled in the art.
[0027] Preferably, the compositions according to the invention are in a form suitable for topical application. These compositions must therefore contain a physiologically acceptable medium, that is to say, compatible with the skin and integuments, without risk of discomfort during their application and cover all suitable cosmetic forms. By topical application is meant the application or spreading of the soft Schinus extract according to the invention, and more particularly a composition containing it, on the surface of the skin or mucosa. The skin refers more particularly to the skin of the face, in particular the outline of the eyes and the mouth, the nose, the forehead, the neck, the hands, as well as the skin of the rest of the body.
[0028] The compositions for carrying out the invention may in particular be in the form of an aqueous, hydro-alcoholic or oily solution, an oil-in-water emulsion, water-in-oil or multiple emulsions; they may also be in the form of suspensions, or even powders, adapted to an application on the skin, the mucous membranes, the lips and / or the hair.
[0029] These compositions may be more or less fluid and also have the appearance of a cream, a lotion, a milk, a serum, an ointment, a gel or a paste. or foam. They may also be in solid form, as a stick or applied to the skin as an aerosol. As a physiologically acceptable excipient commonly used in the intended field of application, there may be mentioned for example adjuvants necessary for the formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens. , self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc. In all cases, those skilled in the art will ensure that these adjuvants and their proportions are chosen in such a way as not to impair the desirable properties of the composition according to the invention. These adjuvants may, for example, correspond to 0.01 to 20% of the total weight of the composition. When the composition according to the invention is an emulsion, the fatty phase may represent from 5 to 80% by weight and preferably from 5 to 50% by weight relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the composition are chosen from those conventionally used in the field under consideration. For example, they can be used in a proportion ranging from 0.3 to 30% by weight, relative to the total weight of the composition.
[0030] According to another advantageous embodiment of the invention, the Schinus soft extract according to the invention may be encapsulated or included in a cosmetic vector such as liposomes or any other nanocapsule or microcapsule used in the field of cosmetics or adsorbed onto powdery organic polymers, inorganic carriers such as talcs and bentonites.
[0031] Advantageously, the composition according to the invention may comprise, in addition to the active agent according to the invention, at least one other active agent having cosmetic effects similar to and / or complementary to those of the invention. According to the invention, this active agent is defined as an "additional active agent". For example, the additional active agent (s) may be chosen from: anti-aging, firming, lightening, moisturizing, draining, microcirculation promoting agents, pharmaceutical agents, exfoliants, desquamating agents, stimulating the extracellular matrix, activating energy metabolism, antibacterial, antifungal, soothing, anti-radical, anti-UV, anti-acne, anti-inflammatory, anesthetic, providing a feeling of heat, providing a sensation of freshness, slimming. Such additional agents may be chosen from the groups comprising: vitamin A and especially retinoic acid, retinol, retinolpropionate, retinol palmitate; Vitamin B3 and more particularly niacinamide, tocopherol nicotinate; - vitamin B5, vitamin B6, vitamin B12, panthenol; vitamin C, in particular ascorbic acid, ascorbyl glucoside, ascorbyl tetrapalmitate, magnesium and sodium ascorbyl phosphate; vitamins E, F, H, K, PP, coenzyme Q10; metalloproteinase inhibitors, or a TIMP activator; DHEA, its precursors and derivatives; amino acids such as arginine, ornithine, hydroxyproline, hydroxyproline dipalmitate, palmitoylglycine, hydroxylysine, methionine and its derivatives, N-acyl amino acid compounds; Natural or synthetic peptides, including di-, tri-, tetra-, penta- and hexapeptides and their lipophilic derivatives, isomeric and complexed with other species such as a metal ion (eg copper, zinc) , manganese, magnesium, and others). By way of examples, mention may be made of the peptides commercially known under the name MATRIXYL®, ARGIRELINE®, CHRONOGENTM, LAMINIXYL ISTM, PEPTIDE Q1OTM, COLLAXYL TM (patent FR2827170, ASHLANDO), PEPTIDE VINCI O1TM (patent FR2837098, ASHLANDO), PEPTIDE VINCI O2TM (patent FR2841781, ASHLANDO), AlPeptideTM (patent FR2846883, ASHLANDO) or the peptide of synthesis of Arg-Gly-Ser-NH2 sequence, marketed under the name AlPeptideTM by ASHLANDC,; - Artemia sauna extract, marketed under the name GP4GTM (FR2817748, ASHLANDO); plant peptide extracts such as linseed extracts (Lipigenin TM, patent FR2956818, ASHLANDO), extracts of soya, spelled, vine, rapeseed, flax, rice, maize, pea; yeast extracts, for example DynagenTM (patent FR2951946, ASHLANDO) or ActopontineTM (patent FR2944526, ASHLANDO); Dehydroacetic acid (DHA); phystosterols of synthetic or natural origin; salicylic acid and its derivatives, alpha- and beta-hydroxy acids, silanols; amino sugars, glucosamine, D-glucosamine, N-acetyl glucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine; extracts of polyphenols, isoflavones, flavonoids, such as grape extracts, pine extracts, olive extracts; Lipids such as ceramides or phospholipids, oils of animal origin, such as squalene or squalane; vegetable oils, such as sweet almond, coconut, castor oil, jojoba, olive, rapeseed, peanut, sunflower, wheat germ, corn germ, soybean oil , of cotton, alfalfa, poppy, pumpkin, evening primrose, millet, barley, rye, safflower, 3034664 21 of passionflower, hazelnut, palm, apricot kernel, avocado of calendula; ethoxylated vegetable oils, shea butter; - all UV screens and sunscreens; Cyclic AMP and its derivatives, the activating agents of the adenylate cyclase enzyme and the phosphodiesterase enzyme inhibiting agents, the centella asiatica extract, the asiaticoside and the Asian acid, the methyl xanthines, the theine, caffeine and its derivatives, theophylline, theobromine, forskolin, esculin and esculoside, ACE inhibitors, Val-Trp peptide, neuropeptide Y inhibitors, enkephalin, gingko biloba extract, dioscorea extract, rutin, yerba mate extract, guarana extract, oligosaccharides, polysaccharides, carnitine, ivy extract, fucus extract, the hydrolyzed extract of Prunella vulgaris, the hydrolysed extract of Celosia cri stata, the extract of Anogeissus leiocarpus, the extract of leaves of Manihot u tilissima, the palmitoylcarnitine, the carnosine, the taurine, the extract of elderberry , extracts of algae such as the extract of Palmaria Palmata. By way of illustration, an example of a formulation of a cosmetic composition containing a Schinus soft extract obtained according to the invention is given below: Example 3: Care Cream (Reference Formula # TCAAE08) Ingredients (Name of brand) INCI mass Purified water QSP 100 Aqua Phase A Tetrasodium EDTA Glycerin (and) Glyceryl Acrylate / Acrylic Acid Copolymer (and) Laureth-23 (and) Dimethicone Na4 EDTA LubrasilTM II DM hydrogel 0.05 3.00 Phenoxyethanol (and) Methylparaben (and) Ethylparaben (and) Propylparaben Liquapar / RokonsalTM MEP Preservative 1.00 3034664 22 Phase B UltraThix ™ P-100 Acrylic Acid / VP polymer Crosspolymer 0.60 Phase C Sodium Hydroxide Sodium Hydroxide 0.02 Aqua Purified Water 0.50 PhaseD Belsil * W3230 Bis-Stearoxydimethylsilane, 2.00 stearyl alcohol, Dimethicone Simulsol * 165 PEG-100 Stearate (and) Glyceryl 2.00 Stearate Refined Shea Butter Butyrospermum Parkii (Shea) 2.00 Butter CeraphylTM 28 Cetyl Lactate 1.50 CeraphylTM 791 th Sterocetyl Stearoyl Stearate 2.00 CeraphylTM ODS Ester Octyldodecyl Stearate 3.00 CeraphylTM 368 Ester Ethylhexyl Palmitate 4.00 Phase E Sodium hydroxide Sodium Hydroxide 0.03 Purified water Aqua 0.50 Phase F Soft Schinus extract according to Example 1 1.00 Process for preparing the composition of Example 3: 1. Add phase A to the main container and begin homogenizing. Heat to 70-75 ° C. 2. Sprinkle in an Ultralhix ™ P-100 and mix well for about 30 minutes. 3. Add the ingredients from phase D to a separate beaker and heat to 70-75 ° C. 4. Add the pre-mixed phase C to phase A and mix until homogeneous. 5. Maintain at 70-75 ° C and add phase D to the main container and mix well. The emulsion must be homogeneous. 6. Start cooling. 7. At about 50 ° C, add the pre-mixed phase E and mix well. 8. At room temperature, add phase F and mix until uniform. Stop at 25 ° C.
[0032] The composition according to Example 3 is thus in the form of a stable white cream with a pH of between 5.20 and 5.80 and a viscosity (OD) of 25,000-45,000 (Brookfield RVT / Spindle B). RPM / 1 minute / 25 ° C) A so-called "placebo" composition is made in exactly the same way with the exception that the ingredient "Schinus soft extract according to Example 1" is replaced by purified water. This "placebo" composition is more particularly used as a control in certain activity tests hereinafter. According to another aspect, the invention relates to the cosmetic use of a composition comprising in particular an effective amount of a Schinus soft extract obtained according to the invention, to improve the barrier function of the skin and to protect against atmospheric pollutants. . The skin is an organ whose upper part, the stratum corneum and the hydrolipidic film which covers it, has a protective physical barrier role vis-à-vis the external environment. This barrier role, more commonly known as "skin barrier function", is of major importance in tissue homeostasis and more particularly in the protection against pathogenic or non-pathogenic external agents, such as environmental pollutants. However, it is known that alterations of the barrier function result from external stresses such as UV. The consequences are major in the loss of cellular cohesion and mechanical integrity. Some enzymes involved in the terminal phases of keratinocyte differentiation play a key role in UV protection. Changes in the expression and / or activity of these enzymes have important consequences on the integrity of the barrier function and homeostasis of the skin.
[0033] Barrier function alterations are also observed in cases of pathological skin and in cases of healthy older skin due to abnormalities in lipid synthesis most often creating a lack of cell cohesion in the surface layers of the skin. the epidermis may cause an increase in skin permeability. Thus, for example, the lack of cohesion of the cells of the epidermis that occurs during skin aging can result in angular micro-fractures and enlargement of the pores of the skin. Alterations in the barrier function may be due to chemical stress. In this last category, it is necessary to take into consideration environmental air pollutants and more particularly PM2.5 microparticles produced by diesel engines, coal combustion, cigarette smoke. These microparticles of small size are indeed responsible for the majority of the negative effects attributed to pollution due to their capacity to penetrate deeply (Pan TL et al., The impact of urban particulate pollution on skin barrier function and the subsequent drug absorption. Dermatol Sci, 2015). In addition, some of these environmental air pollutants have irritating properties for the skin or can cause inflammatory reactions. However, inflammation may be responsible for increased secretion of sebum (Lefebvre M.A. et al., "Evaluation of the Impact of Urban Pollution on the Quality of Skin: A Multicentre Study in Mexico, Int J Cosmet Sci.
[0034] 2015 Feb 6). On a daily basis, the skin is thus regularly subjected to environmental aggressions (UV, pollution) which can alter the physiological functioning of this barrier organ (Pan TL et al. J Dermatol Sci, 2015).
[0035] The composition according to the invention comprising an effective amount of a Schinus soft extract makes it possible to fight against these external aggressions by promoting an optimal differentiation and cohesion of the cutaneous layers making it possible to maintain a functional histological structure and permeability of the skin. physiological order necessary to maintain this essential barrier function that the skin represents at the level of the tissue and beyond, as a whole. The composition of the organism according to the invention thus contributes to the hydration of the skin and the fight against the appearance of the 10 signs of skin aging. "Signs of skin aging" means any changes in the external appearance of the skin due to aging, such as, for example, wrinkles and fine lines, dark circles, and the tone of internal skin defects that do not translate. systematically by a modified external appearance such as, for example, thinning of the skin, or any internal damage to the skin due to exposure to UV radiation. The composition according to the invention is thus also used to limit the secretion of sebum and reduce the size of the pores of the skin.
[0036] As such, the invention is illustrated hereinafter by the various test results performed. EXAMPLE 4 Effects of a hydro-ethanol extract of Schinus Molle obtained according to Example 2 on the expression of markers of the differentiation, hydration and synthesis of lipids in keratinocytes (in vitro models) Potential effects of a hydro-ethanolic Schinus Molle extract obtained according to Example 2 on cutaneous hydration and barrier function were investigated in a model of normal human epidermal keratinocytes (NHEK), crevices, bags under the eyes , wilting, loss of elasticity and / or skin, dullness or lack of brightness, pigmentary but also any modifications by analyzing the expression profile of specific genes for the differentiation of keratinocytes and the synthesis of lipids by a reverse transcription quantitative polymerase chain reaction (RT-qPCR) method.
[0037] 5 Biological models: Normal human epidermal keratinocytes (NHEK) Cell type: NHEK, used at 3rd pass Conditions of 37 ° C, 5% CO2 culture: Culture medium: KeratinocyteSFM (Serum free medium) supplemented with Epidermal Growth Factor (EGF) 0 25 μg / ml Pituitary extract (EP) 25 μg / ml Gentamycin 25 μg / ml Test medium: Keratinocyte SFM supplemented with Gentamycin 25 μg / ml Compounds tested 10 hydro-ethanol extract of Schinus soft obtained according to Example 2, under powder form, stored at room temperature; 10% stock solution (100 mg / ml) in DMSO (dimethyl sulfoxide); Concentrations tested: 0.0001%; 0.0003% and 0.001% on NHEK. Culture and Treatment Normal human epidermal keratinocytes (NHEK) were inoculated and cultured in culture medium for 24 hours.
[0038] The medium was then replaced with the test medium containing or not (control) the hydroethanolic extract of Schinus soft obtained according to Example 2 tested or the positive reference of differentiation (calcium chloride at 1.5 mM ) and the cells were incubated for 24 hours. All conditions were realized in n = 3. At the end of the incubation, the culture supernatants were removed and the cell mats were rinsed with PBS (phosphate buffered saline) solution. The plates were immediately frozen dry at -80 ° C. Analysis of the differential expression of the markers by PCR array: The expression of the markers was evaluated by RT-qPCR on the mRNAs (messenger RNAs) extracted from the cell mats of each treatment (the replicates were pooled before the extraction of RNA).
[0039] Gene expression analysis was performed in n = 2 using 2 "quantitative PCR array Marker" containing 16 genes, including 2 Housekeeping genes, dedicated to research and adapted to the format. "Screening" (mQPA-NHEKBARRIER-16, produced by BIOalternatives0).
[0040] Reverse Transcription The total RNAs of each sample were extracted using TriPure Isolation Reagent® according to the protocol recommended by the supplier. The quantity and quality of the RNAs were evaluated by capillary electrophoresis (Bioanalyzer 2100, Agilent). Traces of potentially contaminating DNA were removed by treatment with the DNA-free system (Ambion). CDNAs (Complementary Deoxyribonucleic Acids) were synthesized by reverse transcription of total RNA (ribonucleic acids) in the presence of oligo (dT) and the enzyme "Transcriptor Reverse Transcriptase" (Roche ()). The obtained cDNA was quantified by spectrophotometry (Nanovue, GE Healthcare0), and then cDNA amounts were adjusted.
[0041] PCR Quantitative PCR reactions (polymerase chain reactions) were performed by quantitative PCR (Light Cycler, Roche Molecular Systems Inc.) and according to the procedures recommended by the supplier. The reaction mixture (10 μl final) for each sample contained: 2.5 μl of cDNA, the primers of the different markers used, the reaction mixture containing the enzyme Taq DNA polymerase, and the SYBR Green I marker and MgCl 2.
[0042] The raw quantitative PCR data was transferred and processed under Microsoft Excel® software. Fluorescence incorporation into the amplified DNA is measured continuously during PCR cycles. These measurements make it possible to obtain fluorescence intensity curves as a function of PCR cycles and thus to evaluate a relative expression value (ER) for each marker. The number of cycles is determined from the "outlets" of the fluorescence curves. For the same tag analyzed, the later a sample leaves (appears late, after a high number of cycles), the lower the initial copy number of the mRNA. The ER value (relative expression) is expressed in arbitrary units (AU) according to the following formula: (1/2 number of cycles) x 106 The PCR array "mQPA-NHEK-BARRIER-16" contains 2 reference genes (housekeeping genes) used for data normalization. These 2 housekeeping genes, the RP528 (Ribosomal protein S28) and GAPDH (Glyceraldehyde-3-phosphate) genes, are expressed constitutively and their level of expression is not or only slightly affected by the treatments. Thus, the level of expression of each marker is compared to the average expression of these 2 reference genes. Data processing The raw data has been transferred and processed using Microsoft Excel® software.
[0043] 10 Formulas used in this report: Standard error of the mean: esm = standard deviation (Sd) / - Vn The standard error of the mean (esm) represents the deviation of the sample mean from the mean of the true population. The esm is calculated by dividing the standard deviation (Sd) by the square root of the sample size. Results The results are shown in Tables 1 and 2 below. - "CaCl2" complete results (number of cycles and percentage of the control) and effects of the reference (CaC12) on the expression of the different markers studied sorted by "cluster" - "hydro-ethanol extract of Schinus soft obtained according to 25 l Example 2: Complete results (number of cycles and percent of the control) and effects of this compound on gene expression, sorted by "cluster." Tables 1 and 2: Effects of a Schinus hydroethanolic extract According to Example 2, the expression of markers of differentiation, hydration and lipid synthesis in keratinocytes evaluated by a method of RTqPCR was compared with that of mQPA-NHEK-genes. ethanolic soft Schinus obtained according to Example 2 BARRIER-16 n 0.0001% 0.0003% 0.001% Cycle Cycles% Cycles% Indicator Cycles% Indicators s Indicator (Average (Average n HK) HK) (Average HK) Name Complete and 100 100 100 Abbreviation Genes of Household (HK) Ribosomal p rotein S28 RPS28 18.79 19.29 86 19.34 89 19.52 80 18.81 19.29 19.22 19.50 Glyceraldeh yde-3 phosphate dehydrogena se GAPDH 16.77 17.02 103 17.10 103 17.22 105 16.91 17.12 17.10 17.11 Hydration Aquaporin 3 (Gill blood group) AQP3 22.95 23.75 73 23.24 111 22.66 172, transport of solutes 23.06 23.73 23.08 22.58 Extracellular matrix Hyaluronan synthase 3 HAS3 25.15 25.82 78 26.43 59 26.50 54 25.28 25.87 26.15 26.52 Synthesis of lipids Acyl-00A synthetase short- chain family member 2 ACSS2 23.42 23.78 100 23.83 101 23.85 106 23.57 23.78 23.73 23.76 Fatty Acid Synthase FASN 21.30 22.00 77 21.98 80 22.25 69 21.32 21.93 21.88 22.22 Serine palmitoyl transferase SPTLC1 24.19 24.65 85 24.10 129 24.29 115, long chain base subunit 1 24.15 24.71 24.11 24.45 LAG1 homolog, ceramide synthase 6 LASS6 27.52 27.68 110 27.70 108 27.25 157 27.65 27.77 27.87 27.41 UDP-glucose ceramide glucosyltra nsferase UGCG 25.91 25.71 150 26, 19 112 25.96 133 25.93 25.52 25.93 25.86 Sphingomyel in phosphodiests Terase 1, lysosomal acid SMPD1 26.81 28.34 40 28.57 36 28.15 53 26.89 28.60 28.66 28.19 Glucosidase GBA 24.97 25.18 110 25.33 97 25.43 94 , beta; acid (includes glucosylceridase) 25.09 25.15 25.41 25.62 Sulfotransferase family, cytosolic, 2B, member 1 SULT2B1 25.42 26.59 59 25.72 102 25.08 160 25.52 26.43 25.77 25.30 Arachidonate e 12- lipoxygenx ALOX12 27.73 28.93 50 29.31 45 28.59 64 27.57 28.92 28.94 28.79 3034664 31 Differentiation of keratinocytes Claudin 1 CLDN1 22.78 23.50 75 22.71 122 22.21 195 22.84 23.50 22.96 22.28 Filaggrin FLG 25.12 25.51 95 25.21 113 24.94 158 24.98 25 29 25.14 24.64 Involucrin IVI 24.86 25.70 67 25.13 94 25.54 87 24.95 25.82 25.47 25.47 Transglutam TGM1 28.06 29.03 74 28.29 119 27 , 81 182 inase 1 (K epidermal polypeptide type I, proteinglutaminegammaglutamyl transferase) 28.28 28.74 28.15 27.60 mQPA-NHEK-BARRIER-16 Control Genes Table 2: CaC12 - 1.5 mM Cycles Cycles% Control (RPS28)% Control (GAPDH)% Control (Medium HK) Complete name and 100 Abreviation Household genes (HK) Ribosomal protein S28 RPS28 18.79 19.13 100 96 97 18.81 19.13 Glyceraldehyde-3-phosphate dehydrogenase GAPDH 16.77 17.16 104 100 101 16.91 17.06 Hydration, solute transport Aquaporin 3 (Gill blood group) AQP3 22.95 23.18 115 110 111 23.06 23.09 Matrix Hyaluronan synthase 3 HAS3 25 Extracellular synthesis 25,34 25,28 25,42 Synthesis of lipids Acyl-CoA synthetase sport chair family member 2 ACSS2 23,42 23,34 132 127 128 23,57 23,51 Fatty acid synthase FASN 21,30 21.51 109 105 106 21.31 21.51 Serine SPTLC1 24.19 24.58 99 95 96 palmitoyltransfase, long flesh base subunit 1 24.15 LAG1 homolog 24.46, ceramide synthase 6 LASS6 27.52 27.80 107 103 104 27.65 27.83 UDP-glucose ceramide glucosyltransferase UGCG 25.91 25.96 123 118 119 25.93 25.94 Sphingomyelin phosphodiesteras SMPD1 26.81 27.01 113 108 109 e 1, lysosomal acid 26, 89 27.00 Glucosidase, beta; acid (includes glucosylceramidase) GBA 24.97 24.96 128 123 124 25.09 25.04 Sulfotransferase family, SULT2B1 25.42 24.44 254 244 246 cytosolic, 2B, 25.52 24.47 3034664 32 member 1 Arachidonate 12- lipoxygenase ALOX12 27.73 27.18 158 151 152 27.57 27.48 22.78 22.55 Claudin 1 CLDN1 152 145 147 22.84 22.53 25.12 24.79 Filaggrin FLG 157 151 152 24, 98 24.66 24.86 24.42 Differentiation Involucrin IVI 183 175 177 24.95 24.31 keratinocytes Transglutaminase 28.06 27.01 1 (K epidermal polypeptide type I, protein-TGM1 285 274 276 glutamine-gamma-glutamyltrans iron ase) 28,28 26,96 In Tables 1 and 2, we consider that there is stimulation, when the% is greater than or equal to 150. On the contrary, we consider that there is inhibition, when the% is less than or equal to 65%. Incubation of the NHEKs with the 1.5 mM CaC12 positive differentiation reference, increased the expression of the mRNAs encoding markers of lipid synthesis (SULT2B1 and ALOX12) and keratinocyte differentiation (FLG, IVL). and TGM1). These results were expected and validated the trial. The hydroethanolic extract of Schinus molle obtained according to Example 2, at the highest concentration tested (0.001%), stimulated the expression of the CLDN1, FLG and TGM1 keratinocyte differentiation markers. In addition, an increase in the expression of the AQP3 hydration marker and the lipid synthesis markers LASS6 and SULT2B1 was also observed. This has been associated with a decrease in the expression of certain genes involved in lipid synthesis (SMPD1 and ALOX12). At the two lowest tested concentrations (0.0001% and 0.0003%), only the inhibition of SMPD1 and ALOX12 expression was observed.
[0044] Conclusion In the experimental conditions of this study, the results obtained suggest that the hydroethanolic extract of Schinus soft obtained according to Example 2, at the highest concentration tested (0.001%), would have a positive effect on the hydration and the barrier function of the epidermis. EXAMPLE 5 Evaluation of the protective effect of Schinus soft extract according to Example 1 on keratinocytes subjected to environmental stress by PM10-like polluting particles. The purpose of this study is to determine the protective effect of the Schinus soft extract according to Example 1 at 0.1%, on keratinocytes subjected to environmental stress. Air pollution is associated with the presence of fine particles having a size smaller than 10 pM (PM10-like). Measurement of cellular stress is performed by assaying the enzymatic activity of lactact dehydrogenase (LDH) released into the extracellular medium by the cells. LDH is an oxidoreductase enzyme that catalyzes the conversion of pyruvate to lactate. The cells release this enzyme into the extracellular medium when there is an impairment of cellular integrity.
[0045] This enzyme test has been widely used to assess the presence of lesions and toxicity of tissues and cells. Protocol: The normal human keratinocytes are cultured in serum-free medium and then treated with a Schinus soft extract according to Example 1 at 0.1% diluted in the culture medium for 48 hours at a rate of 2 applications per day. The untreated control is carried out under the same conditions. After 48 hours, PM10-like polluting particles are applied to the cells at two concentrations 250 μg / ml or 500 μg / ml for 24 hours with a single application.
[0046] The following day, the extracellular medium is removed in order to measure the enzymatic activity of lactate dehydrogenase using the precise and detailed protocol of the Lactate Dehydrogenase Activity Assay Kit Kit (Sigma-Aldrich, MAK066).
[0047] Each condition is performed in triplicate. Results: The results presented in FIG. 1 obtained in vitro show that the Schinus soft extract according to Example 1 at 0.1% protects, in a highly significant manner, the keratinocytes subjected to stress at the PM10-like particles. doses of 250 μg / ml (-29.6% of released LDH compared to PM10-like treatment 250 μg / ml) or 500 μg / ml (-38.5% of released LDH compared to PM10-like treatment 500 μg / ml ).
[0048] EXAMPLE 6 Evaluation of the protective effect of Schinus soft extract according to Example 1 on keratinocytes subjected to environmental stress by PM2.5-like polluting particles.
[0049] The purpose of this study is to determine the protective effect of the Schinus soft extract according to Example 1 at 0.1%, on keratinocytes subjected to environmental stress. Measurement of cellular stress is achieved by assaying the enzymatic activity of lactact dehydrogenase (LDH) released into the extracellular medium by the cells. LDH is an oxidoreductase enzyme that catalyzes the conversion of pyruvate to lactate. Cells release this enzyme into the extracellular medium when there is an impairment of cell integrity. Air pollution is associated with the presence of fine particles having a size of less than 10 pM (PM10-like) but also the presence of small particles smaller than or equal to 2.5 pM (PM2.5like). Since their sizes are smaller than those of the pores of the skin, PM2.5-like can directly induce damage.
[0050] Protocol: The normal human keratinocytes are cultured in serum-free medium and then treated with a Schinus soft extract according to Example 1 at 0.1% diluted in the culture medium for 48 hours at 2 applications per day. The untreated control is carried out under the same conditions. After 48 hours, PM2.5-like polluting particles are applied to the cells at a concentration of 100 μg / ml for 24 hours with a single application.
[0051] The following day, the extracellular medium is removed to measure the enzymatic activity of lactate dehydrogenase using the precise and detailed protocol of the Lactate Dehydrogenase Activity Assay Kit kit (Sigma-Aldrich, MAK066). Each condition is performed in triplicate.
[0052] Results: The results presented in FIG. 2 obtained in vitro show that the Schinus soft extract according to Example 1 at 0.1% protects, in a highly significant manner, the keratinocytes subjected to PM2.5 particles stress. -like at a dose of 100 μg / ml (-11% released LDH compared to PM2.5-like treatment 100 μg / ml). Example 7: Evaluation of the effect of the soft Schinus extract according to Example 1 on the level of expression of epigenetic markers and mRNAs involved in the skin barrier function. The purpose of this study is to determine the effect of the Schinus soft extract according to Example 1 at 0.1%, on the expression levels of two long non-coding RNAs (epigenetic modulators of the skin): TINCR (Tissue Differentiation-Inducing Non-Protein Coding RNA) and DANCR (Differentiation Antagonizing Non-Protein Coding RNA) as well as the expression levels of mRNA encoding proteins involved in the barrier function of the skin As: E-cadherin (CDH1), Involucrin (IVL), Transglutaminase-1 (TGM1) and Caspase 14 (CASP14) in normal human keratinocytes.
[0053] Protocol: The normal human keratinocytes are cultured in serum-free medium and then treated with a Schinus soft extract according to Example 1 at 0.1% diluted in the culture medium for 48 hours at a rate of 2 applications per day. The untreated control 10 is carried out under the same conditions. The total RNAs are extracted using the miRVana extraction kit (Ambion, AM1561), then reverse transcription is performed using the High Capacity cDNA reverse transcription kit (Applied Biosystem, 4374966) for each of the 15 conditions. Real-time quantitative PCR is performed using DANCR-specific TaqManC, Gene Expression Assays (Applied Biosystem: 4426961; Hs03653830 g1), TINCR, TGM1, IVL and CASP14 (Applied Biosystem: 4331182; Hs00542141 m1, Hs01070310 m1, Hs00846307 s1). , Hs00201637 m1) and the TaqManC reaction buffer, Gene Expression Master Mix (Applied Biosystem: 4369514), in the StepOne Plus thermal cycler (Applied Biosystem). TaqManC, 18S-specific Gene Expression Assays is used as endogenous control (Applied Biosystem: 4331182; Hs99999901 s1). The relative quantification of the expression of the different mRNAs is carried out by the comparative method of Ct. The results are given as a percentage of increase of the level of expression relative to the control condition.
[0054] Results (Table) and Conclusions: Skin Barrier Function Biological Markers Biological Markers: Epigenetic MRNA DANCR TINCR E-Involucrin Transglutaminase-Caspase Cadherin 1 14 -26% + 65% + 34% + 283% + 185% + 152% 3034664 The results listed in the Table above show that the Schinus soft extract according to Example 1 at 0.1% positively modulates epidermal differentiation, in vitro. The soft Schinus extract according to Example 1 at 0.1% increases, in vitro, the expression of the mRNAs coding for E-cadherin, involucrin, transglutaminase-1 and Caspase 14, proteins involved in the barrier function of the skin. EXAMPLE 8 Evaluation of the Effect of the Soft Schinus Extract According to Example 1 on the Cellular Cohesion of the Cutaneous Barrier by the Study of Claudine 1. The purpose of this study is to evaluate the effect from Schinus soft extract according to Example 1 at 1% on the expression of the Claudine 1 protein involved in the tight junctions of the epidermis. Claudine 1 forms a gradient of expression in the epidermis. Tight junctions are dynamic structures that ensure cell cohesion and barrier function of the skin.
[0055] Protocol: Human skin biopsies 6 mm in diameter are maintained in culture ex vivo in the presence of a specific medium (DMEM lg / L, HAMF12, SVF and antibiotics) on inserts deposited in 6-well plates. The biopsies are cultured for 48 hours and receive 2 applications per day of Schinus soft extract according to Example 1 diluted to 1% in 1X PBS or 1X PBS for the control condition. The biopsies are then fixed in formaldehyde and then embedded in paraffin. Skin sections of 4 μm thick are then made. Immunolabeling was performed using a rabbit specific polyclonal rabbit antibody (Abcam, ab15098) followed by a fluorochrome-coupled anti-rabbit secondary antibody (Invitrogen, A21206). The biopsies are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Fluorescence quantification using the Volocity® image analysis software (PerkinElmer, Inc.) was performed from the resulting photographs.
[0056] Results and Conclusions: Treatment with Schinus soft extract according to Example 1 at 1% allows to observe a highly significant increase in Claudine 1 expression of 128.2%, compared with the condition control treated. with 1X PBS, for the ex vivo study (Student's t-test, n = 3 +/- wk (wk: standard error of the mean)). The treatment with Schinus soft extract according to Example 1 at 1% would cause a better cohesion of the most differentiated layers of the epidermis and thus enhance the skin barrier function. Example 9 Evaluation of the effect of the Schinus soft extract according to Example 1 on cellular cohesion undergoing the cutaneous barrier function by the study of hyaluronic acid on biopsies ex vivo. The purpose of this study is to evaluate the effect of the Schinus soft extract according to Example 1 at 1% on the expression of hyaluronic acid involved in the establishment of the cutaneous barrier. Hyaluronic acid is abundant in extracellular matrices and also contributes to the hydrodynamics of the epidermis by modulating epidermal differentiation, and improving skin repair.
[0057] Protocol: Human skin biopsies 6 mm in diameter are maintained in culture ex vivo in the presence of a specific medium (DMEM 1 g / L, HAMF12, SVF and antibiotics) on inserts deposited in 6-well plates. The biopsies are cultured for 48 hours and receive 2 applications per day of Schinus soft extract according to Example 1 diluted 1% in 1X PBS or 1X PBS for the control condition. The biopsies are then fixed in formaldehyde and then included in the paraffin. Skin sections of 4 μm thick are then made. The detection of hyaluronic acid is carried out using a biotinylated hyaluronic acid binding protein (Coger-Seikagaki america, 400-763-1A) and then a streptavidin coupled to a fluorochrome (Invitrogen, S32354). The biopsies are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Quantitation of the fluorescence, using the software Volocity® image analysis software (PerkinElmer, Inc.) was performed from the photographs obtained. Results and Conclusions: Treatment with Schinus soft extract according to Example 1 at 1% shows a very significant increase of hyaluronic acid in the dermis and epidermis by 58.9%, compared with the control condition treated with 1X PBS, for the ex vivo study (Student's t-test, n = 3 +/- wk).
[0058] Treatment with Schinus soft extract according to Example 1 at 1% would allow an improvement of the cutaneous barrier function. EXAMPLE 10 Evaluation of the protective effect of the Schinus soft extract according to Example 1 on the lipid content of the keratinocytes in culture. The purpose of this study is to determine the effect of the Schinus soft extract according to Example 1 at 0.1%, on the lipid content of keratinocytes, which underlies the cutaneous barrier function. Protocol: The normal human keratinocytes are cultured in serum-free medium and then treated with a Schinus soft extract according to Example 1 at 0.1% diluted in the culture medium for 48 hours at 2 applications per day. The untreated control is carried out under the same conditions. After 48 hours, the cells are fixed with 3.7% formaldehyde for 10 minutes (Sigma-Aldrich, F1635). After rinsing with PBS, the cells are contacted with Nile Red at 100 nM for 10 minutes (Sigma-Aldrich, N3013) before being rinsed with 1X PBS. The cells are then examined under an Epifluorescence microscope (Zeiss Axiovert 200M microscope). Quantitation of the fluorescence using the Volocity® image analysis software (PerkinElmer, Inc.) was performed from the resulting photographs. Results: The microscopic observations show a highly significant fluorescence with the Schinus soft extract according to Example 1 at 0.1%, on the synthesis of the lipids of the keratinocytes (+ 102% compared to the untreated (according to the t test of Student, n = 3 +/- sem)). Treatment with Schinus soft extract according to Example 1 at 0.1% would make it possible to strengthen the cutaneous barrier function. Example 11: Evaluation of the effect of Schinus soft extract according to Example 1 on the cellular cohesion of the cutaneous barrier by the study of lipids of the epidermis on ex vivo biopsies. The purpose of this study is to evaluate the effect of Schinus soft extract according to Example 1 to 1% on the lipid content of the epidermis. The lipids composing the epidermis have an important role in maintaining the barrier function and in protecting it against external aggressions.
[0059] Protocol: Human skin biopsies 6 mm in diameter are maintained in culture ex vivo in the presence of a specific medium (DMEM 1 g / L, HAMF12, SVF and antibiotics) on inserts deposited in plates 6. well. The biopsies are cultured for 48 hours and receive 2 applications per day of Schinus soft extract according to Example 1 diluted to 1% in 1X PBS or 1X PBS for the control condition. The biopsies are then fixed in formaldehyde and then included. in paraffin. Skin sections of 4 μm thick are then made. Detection of the lipid content of the epidermis is carried out using a solution of 100 nM Nile Red (Sigma-Aldrich, N3013) and then incubation in an alkaline buffer. The biopsies are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Quantitation of the fluorescence, using the software Volocity® image analysis software (PerkinElmer, Inc.) was performed from the photographs obtained.
[0060] Results and Conclusions: Treatment with Schinus soft extract according to Example 1 at 1% shows a very significant increase in lipids in the epidermis by 41%, compared to the control condition treated with PBS. 1X, for ex vivo study (Student's t-test, n = 3 +/- wk). Treatment with Schinus soft extract according to Example 1 at 1% would strengthen the skin barrier function.
[0061] Example 12: Evaluation of the effect of Schinus soft extract according to Example 1 on the barrier function of the skin after SDS stress.
[0062] The purpose of this study is to evaluate, in vitro, on reconstructed tissues, the effect of Schinus soft extract according to Example 1 at 1%, on the permeability of the skin after SDS stress. using a fluorescent dye. Reconstructed human epidermis (RHE) is treated, from day 10 post-reconstruction, topically with a Schinus soft extract according to Example 1 diluted to 1% in 1X PBS for 48 hours at 2 applications per day. day. The control is treated with 1X PBS under the same conditions.
[0063] The RHEs are then treated, on the 12th day post-reconstruction, with 0.15% SDS for 3 hours and then rinsed 5 times with 1X PBS. The RHEs are then treated with 1 mM lucifer yellow (Fluka, 62644), fluorescent dye, for 2 hours. The RHEs are rinsed with 1X PBS and then fixed in formaldehyde and then embedded in paraffin. Skin sections of 4 μm in thickness are made and deposited on a slide. The slides are then deparaffinized and passed through several successive alcohol baths. Penetration of the dye through the barrier and into the tissue is observed under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Diffusion of fluorescence across the epidermis is evaluated from photographs obtained using the software Volocity® image analysis software (PerkinElmer, Inc.).
[0064] Results and Conclusions: The RHE pre-treated with a Schinus soft extract according to Example 1 at 1% and stressed with 0.15% SDS, leave less penetrate the dye. A 16% significant decrease in dye diffusion in the epidermis compared to the 1X PBS condition and stressed with 0.15% SDS was observed (Student's t-test, "SDS 0.15%" n = 4). , "1% + SDS" n = 7, mean ± sem). Treatment with Schinus soft extract according to Example 1 at 1% would make it possible to strengthen the cutaneous barrier function.
[0065] EXAMPLE 13 Effect of a Soft Schinus Extract Formulated According to Example 3 on the Regulation of Sebum Secretion - In Vivo Study Objective: In vivo study of the effect of a Schinus soft extract formulated according to Example 3 Example 3 on the regulation of the secretion of sebum against the placebo composition. Number of volunteers: 10 (24 to 46 years old).
[0066] Study: double blind placebo study. Products tested: Schinus soft extract, formulated at 1% in a composition (Reference Formula # TCAAE08) according to Example 3 versus the placebo composition.
[0067] Application number: 2 applications per day, morning and evening, at a dose of 2 mg / cm 2. Duration of the test: 3 weeks. Control visits: OJ, J0-t1h, J7 and J21.
[0068] Measurements: The secretion of sebum is evaluated by the application of SebutapeO, an opaque white adhesive permeable to sebum, on the skin of the volunteers. The areas that are in contact with the sebum form transparent "spots". The quantitative analysis of the surface and the number of spots present on Sebutapes is carried out using Quantiseb® software. Pore size is independently assessed on photos using a DermaTop®.
[0069] Statistical Analyzes: Student's "t" test. Results: After application of the composition (reference # TCAAE08) comprising a Schinus soft extract formulated at 1% according to Example 3, the analysis shows a statistically significant decrease at J7 and J21 of the surface of the spots (FIG. 3 and Table 3) and the number of spots / cm 2 (Figure 4 and Table 4). Table 3: Evolution of sebum secretion (measurement of spot area) after application of the composition containing 1% Schinus soft extract according to Example 3 Treatment Mean Time (mm 2) Sem P% of% change volunteers having decreased sebum secretion Placebo J7-J0 3.26 0.97 0.0004 *** -81% 100 Formula reference J7-J0 -3.59 1.1 # TCAAE08 (Schinus soft extract at 1% according to Example 3) Placebo J21-J0 3.04 1.19 0.0051 ** -72.6% 90 Reference Formula J21-J0 -3.09 1.46 # TCAAE08 (Schinus molle extract at 1 % according to example 3) **: very significant; ***: highly significant with the Student's test; 10 mean n = 10 +/- wk (standard error of the mean). Table 4: Evolution of sebum secretion (measurement of the number of spots) after application of the composition containing the 1% Schinus soft extract according to Example 3 Treatment Time Average sem P% of% of (number / cm 2 ) change-willingly having had a decrease in the number of spots Placebo J7-J0 50.32 15.09 0.0061 ** -40% 90 Formula reference J7-J0 -24.06 14.58 # TCAAE08 ( 1% Schinus soft extract according to example 3) Placebo J21-J0 45.68 11.23 0.0033 *** -37.2% 80 Reference formula J21-J0 -22.96 16.86 # TCAAE08 (Extract. of Schinus soft to 1% according to example 3) **: very significant; ***: highly significant with the Student's test; average n = 10 +/- wk (standard error of the mean).
[0070] Furthermore, the photos of DermaTop (not shown) show a visible effect on the reduction of the pore size and of their number as early as 1 hour after application (J0-t1h) of the composition (formula # TCAAE08) comprising a Schinus soft extract according to Example 3 compared to placebo. EXAMPLE 14 Effect of a Schinus Soft Extract Formulated According to Example 3 on the Appearance of Wrinkles - In Vivo Study Purpose: Study of the Effect of a Schinus Soft Extract Formulated According to Example 3 on the appearance of wrinkles against placebo composition. Number of volunteers: 14 including 4 test outings. The test was analyzed on 10 volunteers aged 52 to 64 years.
[0071] Study: double blind placebo study. Products tested: Schinus soft extract, formulated at 1% in a composition (Reference Formula # TCAAE08) according to Example 3 versus the placebo composition.
[0072] Application number: 2 applications per day, morning and evening, at a dose of 2 mg / cm 2. Duration of the test: 8 weeks. Control visits: OJ and J56.
[0073] Measurements: The number and depth of wrinkles are analyzed on silicone replicas made on the crow's feet using Quantirides® software. The visual appearance is assessed by color photos of the crow's feet using the HeadScanC photo bench. Statistical analyzes: depending on the normality of the data distribution, Student's "t" test or Wilcoxon test were used.
[0074] Results: After application of the composition (reference # TCAAE08) comprising a Schinus soft extract formulated at 1% according to Example 3, there was a statistically significant decrease at J56 in the number of wrinkles (FIG. 5 and Table 5). ) and mean wrinkle depth (Figure 6 and Table 5) compared to placebo. Table 5: Study of the effect of a Schinus soft extract formulated according to Example 3 on the number of wrinkles and their average depth Treatment Time Average wk% of% change voluntarily improved Number of Placebo J56 -J0 13.9 10.39 0.0304 * -21.16 70% (7/10) wrinkles Formula J56-J0 -10.8 15.35 reference # TCAAE08 comprising an extract of Schinus soft according to Example 3, formulated in 1% Depth Placebo J56-J0 10.28 8.33 0.0068 ** -22.29 80% (8/10) average (Pm) Formula J56-J0 -6.59 4.4 reference # TCAAE08 including an extract soft Schinus according to Example 3, formulated at 1% *: significant; **: very significant; with the Student's test or the Wilcoxon test (test selected according to the normality of data distribution); average n = 10 +/- wk (standard error of the mean). In addition, the color photos (not shown) show an apparent reduction in wrinkles at day 56 after application of the composition (reference # TCAAE08) comprising a Schinus soft extract according to Example 3, formulated at 1% compared to placebo.
[0075] EXAMPLE 15 Test protocol of a Schinus soft extract according to Example 3, formulated at 1%: Moisturizing effect - In vivo study Objective: study of the moisturizing effect of a Schinus soft extract according to US Pat. Example 3, formulated at 1% against the placebo composition. Number of volunteers: 10 (aged 24 to 40 years). Study: double blind study against placebo.
[0076] 10 Products tested: Schinus soft extract, formulated at 1% in a composition (Reference Formula # TCAAE08) according to Example 3 versus the placebo composition. Number of applications: 1 application at a dose of 2 mg / cm2.
[0077] Test duration: 3h. Visits of control: TO, T3Omin, 11h, 11h30, 12h, 12h30, 13h. Measurements: quantitative measurement of hydration using a corneometer® CM825 corneometer and visual evaluation by expert and volunteers. Statistical analyzes: depending on the normality of the data distribution, Student's "t" test or Wilcoxon test were used.
[0078] Results: A significant increase in hydration (Figure 7 and Table 6) is observed as early as 30 minutes after the application of the composition (reference # TCAAE08) comprising a Schinus soft extract according to Example 3, formulated at 1 %, compared to placebo.
[0079] Table 6: Study of the evolution of skin hydration after application of a Schinus soft extract formulated according to Example 3 Treatment Time Average wk% of (UA) improved volunteers placebo T3Omin-T0 10, 43 1.26 0.0107 * 80% (8/10) Reference Formula 14.43 1.17 # TCAAE08 (Example 3) Placebo Tlh-T0 9.6 1.38 0.0190 * 70% (7/10) Reference Formula 13.53 1.69 #TC AAE08 (Example 3) Placebo T1h30-T0 9.2 1.16 0.0263 * 80% (8/10) Reference Formula 12.97 1.44 # TCAAE08 (Example 3) Placebo T2h-T0 9.4 1.44 0.0352 * 70% (7/10) Formula reference 12.7 1.23 # TCAAE08 (example 3) Placebo T2h30-T0 9.1 1.02 0.0103 * 80 % (8/10) Formula reference 12.87 1.22 # TCAAE08 (example 3) Placebo T3h-T0 8.53 1.22 0.0534n '70% (7/10) Formula reference 11.03 1.64 # TCAAE08 (example 3) 5 ns: not significant; *: significant with the Student's test or the Wilcoxon test (test selected according to the normality of data distribution); average n = 10 +/- wk (standard error of the mean).
[0080] In addition, the calculation of the areas under curves (AUC) shows a significant moisturizing effect over the duration of the study (Figure 8 and Table 7). Table 7: Study of the Moisturizing Effect of a Soft Schinus Extract Formulated According to Example 3 Treatment AUC Sem P% of Percent Enhanced Voluntary Pos Placebo 1560 189.24 Reference Formula 0.0129 * 38.49% 70 % (7/10) # TCAAE08 2160.5 211.43 *: significant with the Student's test; AUC n = 10 +/- wk (standard error of the mean).
[0081] Of course, the invention is not limited to the embodiments and the examples presented above and the person skilled in the art, by means of routine operations, may be led to carry out other non-conventional embodiments. described explicitly, which fall within the broad scope of the invention.

权利要求:
Claims (14)
[0001]
REVENDICATIONS1. A hydro-alcoholic extract of the aerial parts of Schinus molle characterized in that it comprises mainly quercitrin and miquelianin among the polyphenols extracted, in a concentration of at least 0.04% of quercitrine by weight of the total weight of the extract and at least 0.02% of miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently of each other being present in the form of traces.
[0002]
2. Soft Schinus extract according to claim 1, characterized in that it is obtained from the fresh or dried aerial parts of soft Schinus by solid-liquid extraction, with stirring, of 0.1% to 20% by weight of Solid material relative to the total weight used in a hydro-alcoholic solvent, the alcohol being chosen from ethanol, glycerol or propanediol, in a proportion of between 10% and 90% by weight of alcohol per relative to the total weight of the solvent, at a temperature of between 4 and 100 ° C. for a period of between 10 minutes and 4 hours, and by separation of the liquid and solid phases so as to eliminate the solid phase and recover a Schinus extract soft liquid.
[0003]
3. Extract according to claim 2, characterized in that, when the alcohol is ethanol, the liquid Schinus soft extract obtained is dried so as to obtain a solid Schinus soft extract.
[0004]
4. Extract according to one of the preceding claims, characterized in that the aerial parts are dried and crushed. 3034664 51
[0005]
5. Extract according to one of the preceding claims, characterized in that the hydro-alcoholic solvent is a water / alcohol mixture in a proportion of between 50% and 80% by weight of alcohol relative to the total weight of the solvent.
[0006]
6. Process for obtaining a Schinus soft extract according to one of claims 1 to 5, characterized in that it comprises the following steps according to which: a) the aerial parts of Schinus molle are collected; b) the aerial parts of Schinus soft collected, fresh or dried, are dispersed in a proportion of 0.1% to 20% by weight of solid matter relative to the total weight used, in a hydro-alcoholic solvent, an alcohol being chosen from ethanol, glycerol or propanediol, in a proportion of between 10% and 90% by weight of alcohol relative to the total weight of the solvent; c) a solid-liquid extraction is carried out, with stirring, at a temperature between 4 and 100 ° C for a period of between 10 minutes and 4 hours; D) separating the liquid and solid phases so as to eliminate the solid phase and recovering a liquid alcoholic Schinus molle extract comprising predominantly quercitrin and miquelianin among the extracted polyphenols, in a concentration of 60%, respectively; less than 0.04% of quercitrin by weight of the total weight of the extract and at least 0.02% of miquelianine by weight of the total weight of the extract, the other extracted polyphenols taken independently of each other being present in the form of traces; and 65 e) optionally, when the alcohol is ethanol, the liquid Schinus soft extract obtained is dried so as to obtain a solid Schinus soft extract. 3034664 52
[0007]
7. Process according to claim 6, characterized in that the liquid Schinus soft extract obtained is purified by microfiltration, ultrafiltration and / or nanofiltration to concentrate the extract in quercitrin and miquelianine with respect to proteins and polysaccharides also extracted. 75
[0008]
8. Cosmetic composition, characterized in that it comprises, as protective active agent, an effective amount of a Schinus soft extract according to one of claims 1 to 5, and a physiologically acceptable excipient.
[0009]
9. Composition according to claim 8, characterized in that the Schinus soft extract is present in the composition at a concentration of 0.001 to 1% by weight relative to the total weight of the composition, preferably from 0.01 to 1. %.
[0010]
10. Composition according to one of claims 8 or 9, characterized in that it is formulated to be applied topically to the skin.
[0011]
11.The cosmetic use of a composition according to one of claims 8 to 10, for improving the barrier function of the skin and protect against atmospheric pollutants. 95
[0012]
12.Use according to claim 11, for moisturizing the skin and to fight against the appearance of the signs of skin aging. 100
[0013]
13.Use according to claim 11 for reducing the pore size of the skin. 3034664 53
[0014]
14.Composition according to one of claims 8 to 10 for its use to limit the secretion of sebum.

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优先权:

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申请号 | 申请日 | 专利标题

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FR1553082A|FR3034664B1|2015-04-09|2015-04-09|HYDRO-ALCOHOLIC EXTRACT OF SCHINUS MOLLE, COSMETIC COMPOSITIONS COMPRISING SAME AND THEIR COSMETIC USES|

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FR1553082|2015-04-09|FR1553082A| FR3034664B1|2015-04-09|2015-04-09|HYDRO-ALCOHOLIC EXTRACT OF SCHINUS MOLLE, COSMETIC COMPOSITIONS COMPRISING SAME AND THEIR COSMETIC USES|

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EP16713951.8A| EP3280497B1|2015-04-09|2016-04-05|Hydroalcoholic extract ofschinus molle, cosmetic compositions comprising the same and cosmetic uses thereof|

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US15/564,537| US11045669B2|2015-04-09|2016-04-05|Hydroalcoholic extract of Schinus molle, cosmetic compositions comprising the same and cosmetic uses thereof|

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PCT/EP2016/057457| WO2016162343A1|2015-04-09|2016-04-05|Hydroalcoholic extract of schinus molle, cosmetic compositions comprising the same and cosmetic uses thereof|

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CN201680020103.1A| CN107809998B|2015-04-09|2016-04-05|Hydroalcoholic extract of mastic from the animal of the genus mastic, cosmetic composition containing it and cosmetic use thereof|

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HK18104904.7A| HK1245631A1|2015-04-09|2018-04-16|Hydroalcoholic extract of schinus molle, cosmetic compositions comprising the same and cosmetic uses thereof|