Extract of the top growth part of holy basil and cosmetic or dermatological composition containing same

Abstract

The present invention relates to an extract of the top growth of holy basil, in particular of Ocimum sanctum, a method for preparing said extract and an extract obtained by said method. The present invention also relates to a composition comprising such an extract, advantageously a cosmetic, pharmaceutical or dermatological composition. The present invention also relates to such a composition or such an extract for use in the prevention or treatment of disorders or diseases of the skin, mucous membranes or skin appendages, and for use in the prevention or treatment of vascular disorders. Finally, the present invention relates to a method for the cosmetic care of the skin, skin appendages or mucous membranes, with the aim of improving their condition or appearance, which method comprises administering such a composition or such an extract.

Description

[Technical field]

[0001] The present invention relates to a composition comprising an extract of the aerial parts of green or purple holy basil (Ocimum sanctum). The composition is advantageously for cosmetic, pharmaceutical or dermatological purposes. The present invention also relates to a method for extracting an extract of the aerial parts of holy basil, and to the extract obtained by said method. The present invention also relates to such a composition or extract for use in the prevention or treatment of disorders or lesions of the skin, mucous membranes or skin appendages, and in the prevention or treatment of vascular disorders. Finally, the present invention relates to a method for the cosmetic care of the skin, skin appendages or mucous membranes, with the aim of improving their condition or appearance, which comprises administering such a composition or such an extract.

[0002] Holy Basil Plant description Ocimum sanctum (or Ocimum tenuiflorum), commonly known as "holy basil" or "holy basil", is an aromatic plant in the Lamiaceae family. Ocimum sanctum grows to a height of 30-60 cm and has hairy stems. Depending on the variety, the leaves are green or purple. The leaves are simple, have petioles, and are up to 5 cm long. The small, purplish flowers are arranged in elongated racemes. There are two varieties of holy basil: green basil and purple basil. Both varieties have a similar phytochemical composition.

[0003] Holy basil is cultivated in Asia for religious and medicinal uses, particularly in Ayurvedic medicine where it is considered an elixir of life. Holy basil leaves are used in Thai cooking and as an insect repellent.

[0004] phytochemistry Composition of Ocimum sanctum: Protein: 174.5g / kg dry plant Fiber: 90.7g / kg dry plant Ash: 135.6g / kg dry plant

[0005] Many molecules have been identified in the aqueous and alcoholic extracts of Ocimum sanctum. Polyphenols are widely present, with the main compound being rosmarinic acid. Ursolic acid, apigenin derivatives and phenolic acids have also been widely reported.

[0006] The sugars consist of xylose and polysaccharides. The stems and leaves contain saponins, flavonoids, triterpenoids and tannins.

[0007] The mineral composition shows that potassium dominates (see Table 1). [Table 1] Summary of the Invention

[0008] The subject of the present invention is a polyphenol-rich extract of holy basil, in particular a polyphenol-rich extract of the aerial parts of holy basil.

[0009] In the context of the present invention, the terms "holy basil" and "ocimum sanctum" have the same meaning and can be used interchangeably. There are two varieties of holy basil: green and purple. Unless otherwise specified, for the purposes of the present invention, "holy basil" or "ocimum sanctum" refers to the two existing varieties, namely, the green and purple varieties.

[0010] In the context of the present invention, the "aerial parts" of Ocimum sanctum include leaves, stems and flowers, preferably leaves and stems.

[0011] "Polyphenol-rich extract" refers to an extract that contains primarily or essentially polyphenols.

[0012] Thus, the extract of the invention contains at least 6% by weight of polyphenols relative to the total weight of the dry extract, i.e. at least 1.2 mg of polyphenols (calculated as gallic acid) per mL of liquid extract. Advantageously, the extract of the invention contains at least 9% by weight, more advantageously at least 11% by weight, more advantageously at least 15% by weight of polyphenols relative to the total weight of the dry extract. Advantageously, the extract of the invention contains between 6% and 17% by weight, more advantageously between 9% and 17% by weight, more advantageously between 11% and 17% by weight, more advantageously between 15% and 17% by weight of polyphenols. The percentages are expressed relative to the total weight of said dry extract (before the addition of the drying medium) and are determined according to the Folin Ciocalteu method.

[0013] For the purposes of the present invention, polyphenols are molecules that have at least two hydroxyl functional groups carried by one or more benzene rings. Particularly advantageously, polyphenols according to the present invention comprise or consist of hydroxycinnamic derivatives and flavonoids.

[0014] Advantageously, the extract of the invention comprises at least 5% by weight of hydroxycinnamic derivatives, more advantageously at least 7% by weight of hydroxycinnamic derivatives, i.e. at least 1.0 mg of hydroxycinnamic derivatives per mL of liquid extract. More advantageously, the extract of the invention comprises at least 10% by weight, more advantageously at least 15% by weight of hydroxycinnamic derivatives. Advantageously, the extract of the invention comprises between 5% and 20% by weight, more advantageously between 7% and 20% by weight, more advantageously between 10% and 20% by weight, more advantageously between 15% and 20% by weight of hydroxycinnamic derivatives. The percentages are expressed as chlorogenic acid equivalents relative to the total weight of said dry extract (before possible addition of drying medium) and are determined according to the Arnow method.

[0015] Advantageously, the extract of the invention comprises at least 2.5% by weight, expressed as rutin equivalent, and more advantageously at least 3.5% by weight, of flavonoids, relative to the total weight of said dry extract. Advantageously, the extract of the invention comprises between 2.5% and 6% by weight, and more advantageously between 3.5% and 6% by weight, expressed as rutin equivalent, of flavonoids, relative to the total weight of said dry extract. The flavonoid content is determined by the concentration of Loots (AlCl 3 ) method is used to measure.

[0016] Particularly advantageously, the extract according to the invention contains at least 6% by weight of polyphenols relative to the total weight of the dry extract, i.e. at least 1.2 mg of polyphenols (expressed as gallic acid) per ml of liquid extract, in particular at least 5% by weight of hydroxycinnamic derivatives relative to the total weight of the dry extract, and at least 2.5% by weight of flavonoids, expressed as rutin equivalents, relative to the total weight of the dry extract.

[0017] In the context of the present invention, hydroxycinnamic derivatives are molecules which contain the following motif in their structure: [ka] It is understood that the hydrogens of the above motifs may be replaced by any other groups.Hydroxycinnamic derivatives within the meaning of the present invention include in particular hydroxycinnamic acids, esters and glycosylated derivatives.

[0018] Examples of hydroxycinnamic derivatives according to the present invention include cichoric acid, caffeic acid, rosmarinic acid or caffeic acid, and mixtures thereof.

[0019] In the context of the present invention, flavonoids are defined as molecules that contain the following motif in their structure: [ka] It is understood that the hydrogen of this moiety may be replaced with any other group. The dotted lines in the above formula represent single or double bonds.

[0020] Examples of flavonoids according to the present invention include apigenin, apigenin-7-glucuronide or luteolin-7-glucuronide, and mixtures thereof.

[0021] For the purposes of the present invention, the flavonoids and hydroxycinnamic derivatives are typically polyphenols.

[0022] Particularly advantageously, the extract of the invention is essentially free of eugenol and methyleugenol, which are allergenic compounds and which are phenols, not polyphenols, in the sense of the invention. Eugenol and methyleugenol, in particular due to their physicochemical properties, are not classified in the polyphenol family. In fact, these compounds are one of the main components of holy basil essential oil and, unlike the polyphenols of the invention, are steam entrainable compounds.

[0023] Thus, advantageously, the extract of the invention is free of eugenol and methyleugenol or contains less than 50 ppm of eugenol and less than 100 ppm of methyleugenol.

[0024] Advantageously, according to the invention, the extract is not an essential oil, since the components of the extract are not steam entrainable.

[0025] In the context of the present invention, advantageously, the above-mentioned polyphenol-rich extract is obtained by solid / liquid extraction of the aerial parts of Holy Basil in a solvent selected from water, glycol or glycerol, and mixtures thereof. The extraction solvent in the present invention is a non-toxic solvent, more particularly selected from a binary mixture of water / glycol or water / glycerol, advantageously with a proportion of glycol or glycerol in water of 30% to 90%, typically 50% to 80%, more advantageously 60% to 70%, by volume relative to the total volume of the solvent used (water+glycol or water+glycerol). In particular, the extraction solvent is selected from a binary mixture of water / propanediol, water / propylene glycol, water / glycerin, in particular water / propanediol, more particularly water / 1,3-propanediol or water / glycerin.

[0026] Advantageously, the abovementioned glycols are glycols of vegetable origin, in particular propanediol and / or propylene glycol, more particularly propanediol, in particular 1,3-propanediol.

[0027] Advantageously, the abovementioned glycerol is of vegetable origin.

[0028] Another object of the present invention is a process for preparing an extract of the aerial parts of Holy Basil rich in polyphenols, comprising at least one step of solid / liquid extraction in a suitable solvent mixture, in particular as defined in the paragraph above, under optimal conditions of pH, time and temperature known to the person skilled in the art.

[0029] Advantageously, according to the invention, the method for preparing an extract of the aerial parts of Holy Basil rich in polyphenols comprises the following successive steps: a) crushing the above-ground parts of holy basil; b) extracting the ground aerial parts in a solvent selected from water, glycol, glycerol and mixtures thereof, advantageously in a binary mixture of water / glycol or water / glycerol, under optimal conditions of time, pH and temperature; c) separating the solid phase from the liquid phase by decantation and / or centrifugation and / or continuous filtration; and d) Optionally, drying the extract obtained in step c).

[0030] Particularly advantageously, according to the invention, the process for the preparation of the extract comprises a step of removing eugenol or methyleugenol or substantially reducing the eugenol and methyleugenol content, typically by vacuum evaporation or deodorization.

[0031] The crushing of the plant in step a) can be carried out by methods known to those skilled in the art, in particular using a cutter mill, a hammer mill, or the like.

[0032] In step b), the extraction is carried out in the presence of an extraction solvent, preferably chosen from a binary mixture of water / glycol or water / glycerol. Advantageously, the extraction solvent used in step b) is chosen from a binary mixture of water / glycol or water / glycerol, in which the proportion of glycol or glycerol in water is between 30% and 90%, typically between 50% and 80%, more advantageously between 60% and 70%, by volume relative to the total volume of the solvent used (water+glycol or water+glycerol). In particular, the extraction solvent is chosen from a binary mixture of water / propanediol, water / propylene glycol or water / glycerin, in particular water / propanediol, more particularly water / 1,3-propanediol or water / glycerin.

[0033] Advantageously, the glycol is a glycol of vegetable origin, in particular propanediol and / or propylene glycol, more particularly propanediol, in particular 1,3-propanediol. Advantageously, the glycerol is of vegetable origin.

[0034] Furthermore, the solid / liquid extraction in step b) is preferably carried out at a temperature between 20°C and 90°C, in particular between 30°C and 80°C, more in particular between 30°C and 75°C, typically at 60°C.

[0035] In contrast to other prior art methods, the advantage is that the extraction period is short, advantageously between 30 minutes and 4 hours, in particular between 1 and 3 hours, advantageously about 2 hours.

[0036] The separation of the solid phase from the liquid phase in step c) is carried out by methods known to those skilled in the art, in particular by decantation, centrifugation and / or successive filtration, preferably by filtration. During step c), the liquid phase obtained is advantageously purified and concentrated, for example by ultrafiltration and / or sterile filtration. Step c) is carried out until the liquid is completely clear and microbiologically clean to the extent of less than 100 CFU / g total bacteria.

[0037] Advantageously, the polyphenol-rich extract of the invention can be stabilized by a drying step d) by methods known to those skilled in the art.

[0038] The drying in step d) can be carried out in the presence of a carrier, such as, for example, maltodextrin or acacia fibre (Fibregum® from CNI), the carrier content typically varying in a ratio ranging from 0 to 80% of carrier relative to the proportion of dry matter obtained in the extract in liquid form.

[0039] The extract can be dried by lyophilization to obtain a final powder, which advantageously contains 30-70% by weight of dry matter from the extract, with the remainder being 100% lyophilized carrier. More advantageously, the final powder contains 50% extract dry matter and 50% lyophilized carrier.

[0040] Advantageously, the method of the invention does not include step d), and therefore the extract obtained is preserved in an extraction solvent, the solvent being in an amount sufficient to exhibit bacteriostatic / bactericidal activity, in particular the solvent being in an amount of 50% by weight or more relative to the total weight of the extract and the solvent.

[0041] The extraction method of the present invention advantageously allows for a simple implementation of an extract enriched in polyphenols in a single main extraction step, i.e. without the need for multiple successive extractions as in fractionation.

[0042] Preferably, and by way of example, the polyphenol-rich extract of the present invention can be obtained according to the following method: i. Crush the above-ground parts of holy basil; ii. 10% (w / w) of the aerial parts was dissolved in a 1,3-propanediol / water mixture (60 / 40) or a glycerin / water mixture (60 / 40); iii. Extraction at 60°C for 2 hours with stirring; iv. Purified by successive filtration steps; v. Advantageously, eugenol and methyleugenol are removed, typically by vacuum evaporation or deodorization; and vi. Sterile filter.

[0043] In the following description, the expression "extract of the invention" is used to refer to the extract as defined above or to an extract obtained by the method of the invention described above, which has the same composition as the extract as defined above.

[0044] Another object of the invention is a composition comprising an extract according to the invention and a water / solvent mixture with a water / solvent ratio (v / v) of 50 / 50 to 0 / 100, advantageously 30 / 70 to 10 / 90, more advantageously 20 / 80, said solvent being chosen from glycols, vegetable glycerins and mixtures thereof, preferably glycols of vegetable origin, and preferably 1,3-propanediol.

[0045] Advantageously, the composition comprises, relative to the total weight of the composition, 0.001 to 30% by weight, advantageously 0.001 to 10% by weight (expressed as the weight of the dry extract relative to the total weight of the composition) of the extract of the invention, and 70 to 99.999% by weight, advantageously 90 to 99.999% by weight of a water / solvent mixture, the water / solvent ratio being 50 / 50 to 0 / 100, advantageously 30 / 70 to 10 / 90, more advantageously 20 / 80, the solvent being chosen from glycols, vegetable glycerins and mixtures thereof, preferably from glycols of vegetable origin, preferably from 1,3-propanediol. According to this aspect of the invention, the solvent is in an amount effective for the physical and microbiological stabilizing action of the composition of the invention, in particular of the extract of the invention.

[0046] Another object of the invention is a composition comprising as active ingredient the polyphenol-rich extract of the aerial parts of Ocimum sanctum according to the invention, optionally including suitable excipients, as defined in the above paragraphs relating to the extract itself and to the extract obtained by the method of the invention.

[0047] Advantageously, the composition is for cosmetic, pharmaceutical or dermatological purposes. Said composition is preferably formulated for external topical administration.

[0048] Advantageously, the composition of the invention comprises between 0.001 and 10% by weight, typically between 0.01 and 5% by weight, of the extract of the invention relative to the total weight of the composition, the weight of the extract being expressed as dry extract.

[0049] The compositions of the present invention may also contain one or more other active ingredients.

[0050] The compositions of the present invention can be formulated in a variety of formulations suitable for topical administration, including, inter alia, creams, emulsions, milks, ointments, lotions, oils, aqueous solutions, hydroalcoholic solutions, glycolic solutions, powders, patches, sprays, shampoos, varnishes or other products for external administration.

[0051] Depending on its nature (cosmetic, pharmaceutical or dermatological), the composition of the invention may contain at least one cosmetically, pharma- ceutical or dermatologically acceptable excipient. In particular, the composition of the invention may also contain at least one cosmetically, pharma- ceutical or dermatologically acceptable adjuvant known to those skilled in the art, selected from surfactants, thickeners, preservatives, fragrances, colorants, chemical or mineral filters, moisturizers, warming agents, etc. Those skilled in the art know how to adapt the formulation of the composition of the invention using their general knowledge.

[0052] The optimal dosage and galenical form of the composition of the present invention can be determined according to criteria generally considered when establishing a pharmacological, dermatological or cosmetic treatment adapted to a patient or animal, such as the age or weight of the patient or animal, the severity of the general condition, tolerance to the treatment, observed side effects, skin type, etc.

[0053] Another object of the invention is the extract according to the invention or the composition according to the invention for use in the prevention and / or treatment of disorders or lesions of the skin and / or mucosa (gums, periodontal tissues, genital mucosa) and / or skin appendages (hair and nails), whether immature, normal or mature / aged, advantageously inflammatory reactions, oxidative reactions, disorders related to endogenous or exogenous stress (psychological stress, stress related to anxiety states, stress related to radical insults related or not related to pollution (in particular chemical or air pollution) and / or stress related to UV or IR exposure), barrier or homeostasis disorders, ageing, in particular chronological ageing and / or photoageing, photosensitized skin and / or mechanical or thermal insults.

[0054] Another object of the invention is the extract according to the invention or the composition according to the invention for use in the prevention and / or treatment of vascular disorders, in particular redness and couperose.

[0055] Another object of the present invention is the extract of the present invention or the composition of the present invention for use in the prevention and / or treatment of disorders related to imbalance and / or general imbalance of the microbiota of the skin and / or mucosa (gingiva, periodontium, genital mucosa) and / or skin appendages, whether immature, normal or mature / aged.Indeed, the extract of the present invention has a protective activity against mechanical, microbial, thermal and free radical insults in the skin and / or mucosa (gingiva, periodontium, genital mucosa) and / or skin appendages and / or their appendages, whether immature, normal or mature / aged.The extract of the present invention acts to protect the microbiota and thus helps to prevent imbalance of the microbiota.The extract of the present invention also helps to stimulate the immune defense and antioxidant system of the skin.

[0056] Another object of the invention is the use of the extract according to the invention or the composition according to the invention for the manufacture of a cosmetic, pharmaceutical or dermatological composition for the prevention and / or treatment of disorders or lesions of the skin and / or mucosa (gums, periodontal tissues, genital mucosa) and / or skin appendages (hair and nails), whether immature, normal or mature / aged, advantageously inflammatory reactions, oxidative reactions, disorders related to endogenous or exogenous stress (psychological stress, stress related to anxiety states, stress related to radical insults related or not related to pollution (in particular chemical or air pollution) and / or stress related to UV or IR exposure), barrier or homeostasis disorders, ageing, in particular chronological and / or photoaging, photosensitized skin and / or mechanical or thermal insults.

[0057] Another object of the present invention is the use of the extract according to the invention or the composition according to the invention for the manufacture of a pharmaceutical, cosmetic or dermatological composition for the prevention and / or treatment of vascular disorders, in particular redness and couperose.

[0058] Another object of the present invention is the use of the extract according to the invention or the composition according to the invention for the manufacture of a pharmaceutical, cosmetic or dermatological composition for preventing and / or treating an imbalance in the microbiota of the skin and / or mucosa (gingiva, periodontal tissue, genital mucosa) and / or the skin appendages and / or of said appendages, whether immature, normal or mature / aged, and / or disorders associated with said imbalance.

[0059] A further object of the present invention is a method for the prevention and / or treatment of disorders or lesions of the immature, normal or mature / aged skin and / or mucosa (gingiva, periodontitis, genital mucosa) and / or skin appendages (hair and nails), advantageously inflammatory reactions, oxidative reactions, disorders related to endogenous or exogenous stress (psychological stress, stress related to anxiety states, stress related to radical insults related or not related to pollution (especially chemical or air pollution) and / or stress related to UV or IR exposure), barrier or homeostasis disorders, ageing, in particular chronological ageing and / or photoageing, photosensitized skin and / or mechanical or thermal insults, which comprises administering, in particular topically, an effective amount of an extract according to the invention or a composition according to the invention to a subject in need thereof.

[0060] Another object of the present invention is a method for preventing and / or treating vascular disorders, in particular redness and couperoses, which comprises administering, in particular topically administering, to a subject in need thereof an effective amount of an extract according to the invention or a composition according to the invention.

[0061] Another object of the present invention is a method for preventing and / or treating imbalances in the microbiota of the skin and / or mucosa (gingiva, periodontal tissues, genital mucosa) and / or skin appendages and / or of said appendages, whether immature, normal or mature / aged, and / or disorders related to said imbalances, comprising administering, in particular topically, an effective amount of an extract according to the invention or a composition according to the invention to a subject in need thereof.

[0062] In particular, the compositions or extracts of the invention are intended to prevent and / or treat inflammatory or irritant reactions, or barrier or homeostasis conditions or disorders, of the immature, normal or mature / aged skin, skin appendages (hair and nails) and / or mucosa (gums, periodontal tissues, genital mucosa).

[0063] Advantageously, the inflammatory or irritant reaction, the disorder or pathology or disorder of the skin barrier or homeostasis is: acne, rosacea erythrocouperose, vascular disorders, in particular redness and couperose, diaper dermatitis, atopic dermatitis, eczema, contact dermatitis, irritant dermatitis, allergic dermatitis, seborrheic dermatitis (scalp dermatitis in newborns), sensitive skin, reactive skin, dry skin (xeroderma), dry skin, reddened skin, erythema of the skin, aged skin or photoaged skin, photosensitized skin, pigmented skin (melasma, post-inflammatory pigmentation, ...), skin with linear skin atrophy, sunburn, chemicals, substances causing physical changes (e.g. stress in pregnant women), irritation by bacterial or fungal substances, ageing of the skin, in particular photoageing, disorders related to radical attack by chemicals or air pollution and / or disorders related to exposure to UV or IR.

[0064] Advantageously, the inflammatory or irritant reaction, disturbance or lesion or disorder of the mucosal barrier or homeostasis is gingivitis (sensitive gums in newborns, hygiene problems due to smoking, etc.) and inflammation of the external or internal male or female genital spheres.

[0065] Advantageously, the inflammatory reaction, irritant reaction, disturbance or lesion or disorder of the skin appendage barrier or homeostasis is brittle nails, fragile nails, weak hair, brittle hair, dry hair, alopecia, seborrheic dermatitis and folliculitis dermatitis.

[0066] Particularly advantageously, the composition or extract of the invention is intended as an anti-ageing agent against skin ageing, in particular against chrono-ageing and photo-ageing, for example linked to exposure to sunlight or to inflammatory phenomena, or acts on inflamed skin.

[0067] In a preferred embodiment, the extract of the present invention comprises - disorders or lesions of the skin and / or mucous membranes and / or skin appendages, preferably disorders related to endogenous or exogenous stress, such as inflammatory reactions, psychological stress, stress related to anxiety states, stress related to radical insults with or without chemical or air pollution, and / or stress related to UV or IR exposure, preferably disorders or lesions selected from sensitive skin, reactive skin and / or reactive mucous membranes, rough skin, inflamed skin, barrier or homeostasis disorders, photosensitized skin, mechanical and / or thermal insults; and / or - Vascular disorders, preferably redness and couperose; and / or Advantageously, the treatment of atopic dermatitis, eczema, occurrence of unpleasant axillary odor, weakening of the skin barrier, acne, psoriasis, sweat gland abscess, folliculitis, neonatal scalp dermatitis, dandruff, pruritus, especially itchy scalp, candidiasis and bacterial vaginosis, as well as imbalances of the microflora. and / or as a therapeutic agent for the prevention and / or treatment of

[0068] Another object of the present invention is the cosmetic use of the extract according to the invention or of the composition according to the invention in the treatment and / or prevention of dehydrated skin; red skin; aged skin; skin aging.

[0069] Another object of the invention is the cosmetic use of an extract according to the invention or of a composition according to the invention for the care of the skin appendages, in particular for the prevention of brittle nails; brittle nails; weak hair; brittle hair; and dry hair.

[0070] The present invention also relates to a method for the cosmetic care of the skin and / or skin appendages and / or mucous membranes, aimed at improving their condition and / or appearance, which consists of administering a composition or extract according to the invention; in particular improving the firmness, elasticity or length of the skin, strengthening the mechanical properties of the skin and mucous membranes, in particular preventing withered, limp, distended and / or sagging skin and / or strengthening and / or restoring the elasticity or firmness of the skin, or treating fine lines and wrinkles.

[0071] In particular, the present invention relates to a method for the cosmetic care of the skin and / or mucous membranes and / or skin appendages, aimed at preventing damage to the skin barrier and its dehydration or at moisturizing the skin and / or mucous membranes, which comprises applying a composition or an extract according to the invention to the skin and / or mucous membranes and / or skin appendages.

[0072] The present invention also relates to a method for the cosmetic care of the skin and / or skin appendages, aimed at preventing ageing, which comprises applying to the skin and / or skin appendages a composition or extract according to the invention.

[0073] The present invention also relates to the cosmetic use of a composition or an extract or a composition according to the invention for moisturizing the skin and / or mucous membranes, for affecting skin elasticity or firmness, in particular for acting on sensitive skin as a tensile or anti-wrinkle agent.

[0074] In particular, the present invention relates to the cosmetic use of the composition or extract of the present invention in the treatment and / or prevention of dehydrated skin; red skin; aged skin; evening out skin tone; reducing skin blemishes or skin aging, especially reducing chronoaging and / or photoaging.

[0075] The cosmetic uses and beauty care methods of the present invention are typically applied to healthy skin or healthy parts of the body and are not therapeutic. [Brief description of the drawings]

[0076] [Figure 1] FIG. 1 shows the measurement of NO production by keratinocytes under cortisol stress. [Diagram 2] Figure 2 shows the effect of Holy Basil extract on cellular bioenergetics of fibroblasts exposed to pollutant stress, specifically Fig. 2A shows the effect on oxygen consumption, Fig. 2B shows the effect on ATP production, Fig. 2C shows the effect on lactate production, and Fig. 2D shows the effect on NAD / NADH ratio (*p<0.05; **p<0.01 vs. untreated control; #p<0.05; ##p<0.01 vs. pollutants). [Diagram 3] FIG. 3 shows the percentage of damaged mitochondria per cell (###p<0.0001 vs. contaminant). [Figure 4] FIG. 4 shows lysosome density (intensity / μm2) (***p<0.0001 vs. control; #p<0.05; ###p<0.0001 vs. contaminant). [Diagram 5] Figure 5. Mitophagy (percentage of lysosomes co-localized with damaged mitochondria) (***p<0.0001 vs. control; ###p<0.0001 vs. contaminant). [Figure 6] FIG. 6 shows the percentage of fibroblast migration between T0 and T40h. EXAMPLES

[0077] Example 1: Extract of the invention The extract of holy basil was obtained by the following method. a) 10% (w / w) of ground aerial parts of Ocimum sanctum (purple cultivar) was dissolved in a 40 / 60 w / w water / propanediol mixture. b) Extracted at 60°C for 2 hours c) Solid / liquid separation was performed by continuous filtration.

[0078] The polyphenol-rich liquid extract thus obtained had the following characteristics (% by weight of molecules / dry extract of the extract obtained): Dry extract (2 hours, 105°C, ventilated oven): 1.0% Total polyphenols: 11.4% Flavonoids: 3.6% Hydroxycinnamic derivatives: 7.6%

[0079] Example 2: Extract of the invention The extract of holy basil was obtained by the following method. a) 10% (w / w) of ground aerial parts of Ocimum sanctum (green variety) was dissolved in a 40 / 60 w / w water / propanediol mixture. b) Extraction was performed at 60°C for 3 hours. c) Solid / liquid separation was performed by continuous filtration.

[0080] The polyphenol-rich liquid extract thus obtained had the following characteristics (% by weight of molecules / dry extract of the extract obtained): Dry extract (2 hours, 105°C, ventilated oven): 1.4% Total polyphenols (measured by Folin Ciocalteu method): 16% Flavonoids: 3.5% Hydroxycinnamic derivatives: 20%.

[0081] Example 3: Biological activity test of the extract of the present invention A. Antioxidant effect A.1. Antioxidant effect A.1.1 H in normal human keratinocytes 2 O 2 Activity against inducible ROS production H 2 O 2 The antioxidant activity of holy basil extract was investigated by assessing the amount of ROS (reactive oxygen species) produced by keratinocytes exposed to induced oxidative stress.

[0082] A. Materials and Methods Normal human keratinocytes were treated for 24 h with six concentrations (0.00032% to 0.01% dm) of holy basil extract obtained according to Example 1 or with antioxidant reference molecules: 10 μM quercetin or 500 μM vitamin C, prior to the uptake of the H2DCF-DA probe.

[0083] The keratinocytes were then soaked in 100 μM hydrogen peroxide (H 2 O 2 ) for 20 min.

[0084] The production of ROS was assessed by measuring the fluorescence emitted by the probe contacted with ROS.

[0085] The results were statistically analyzed using one-way analysis of variance followed by Tukery's test.

[0086] b.Results The results are summarized in Table 2. The holy basil extract dose-dependently inhibited H 2 O 2 It significantly inhibited the production of ROS by keratinocytes in response to induced oxidative stress, thus exhibiting antioxidant activity.

[0087] [Table 2]

[0088] A.1.2. Activity against UVA-induced ROS production in reconstructed human epidermis The protective effect against oxidative stress induced by UVA irradiation was examined by ROS assay on reconstituted human epidermis.

[0089] A. Materials and Methods The reconstructed human epidermis was pretreated for 24 hours with a formulation containing 3% basil extract obtained according to Example 1 or with its placebo. The epidermis was then treated with 30 J / cm 2 The specimens were exposed to UVA.

[0090] The production of ROS was assessed by measuring the fluorescence emitted by the H2DCF-DA probe contacted with ROS.

[0091] b.Results The results are summarized in Table 3. The holy basil extract formulated at 3% inhibited UVA-induced ROS generation, demonstrating a protective effect against oxidative stress induced by UVA irradiation.

[0092] [Table 3]

[0093] 1.3. Activity against menadione-induced ROS production in normal human keratinocytes and fibroblasts The antioxidant activity of holy basil extract was investigated by assessing the amount of ROS produced by keratinocytes or fibroblasts exposed to menadione-induced oxidative stress.

[0094] A. Materials and Methods Normal human keratinocytes on the one hand and normal human fibroblasts on the other hand were treated for 24 hours with 0.001% and 0.005% of dried matter (dm) of the extract of Holy Basil obtained according to Example 1 or with 10 mM N-acetylcysteine ​​(NAC), a ROS scavenger molecule used as an antioxidant reference. Oxidative stress was then induced by incubation for 1 hour in the presence of 50 μM or 20 μM menadione (2-methyl-1,4-naphthoquinone).

[0095] ROS were labeled with CellRox® probes; intracellular fluorescence, which is proportional to the amount of ROS, was measured.

[0096] The results were statistically analyzed using one-way analysis of variance followed by Dunnett's test.

[0097] b.Results The results are shown in Tables 4 and 5. Holy basil extract significantly inhibited menadione stress-induced ROS production in keratinocytes (Table 4) and fibroblasts (Table 5), confirming its antioxidant activity.

[0098] [Table 4]

[0099] [Table 5]

[0100] A.2. Inhibition of lipid peroxidation ROS are the main activators of lipid peroxidation, mainly targeting polyunsaturated lipids bound to cell membranes. The anti-lipid peroxidation activity of oasis extracts was investigated in keratinocytes or fibroblasts exposed to oxidative stress induced by cumene hydroperoxide.

[0101] A. Materials and Methods Normal human keratinocytes on the one hand and normal human fibroblasts on the other hand were treated for 24 hours with 0.001% and 0.005% of dried extract of Holy Basil obtained according to Example 1, or with 200 μM of vitamin E, an antioxidant reference. Oxidative stress was then induced by incubation for 1 hour in the presence of 200 μM or 50 μM of cumene hydroperoxide.

[0102] Lipid peroxidation was revealed by labeling with BODIPY® 581 / 591 C11; intracellular fluorescence was measured.

[0103] The results were statistically analyzed using one-way analysis of variance followed by Tukey's test.

[0104] b.Results The results are summarized in Tables 6 and 7. The oasis extract significantly inhibited oxidative stress-induced lipid peroxidation in keratinocytes (Table 6) and fibroblasts (Table 7), confirming its antioxidant activity.

[0105] [Table 6]

[0106] [Table 7]

[0107] A.3. Effects on antioxidant defense Glutathione (GSH) is the major endogenous antioxidant peptide that can neutralize ROS. Depletion of intracellular GSH contributes to oxidative stress-induced damage associated with mitochondrial toxicity.

[0108] The effect of oasis extract on intracellular glutathione stores was evaluated in fibroblasts exposed to menadione-induced oxidative stress.

[0109] A. Materials and Methods Normal human fibroblasts were treated for 24 hours with 0.001% and 0.005% of the dried extract of Holy Basil obtained according to Example 1, or with 10 mM NAC, an antioxidant reference, and then oxidative stress was induced by incubation for 4 hours in the presence of 25 μM menadione.

[0110] GSH stocks were monitored using the fluorescent probe monochlorobimane (Mbi); intracellular fluorescence was measured, which is proportional to the amount of GSH.

[0111] b.Results The results are summarized in Table 8. The holy basil extract significantly restored intracellular glutathione depleted under menadione-induced oxidative stress compared to the antioxidant reference (10 mM NAC), confirming the antioxidant activity of the extract.

[0112] [Table 8]

[0113] A.4. Conclusion Taken together, these results demonstrate the antioxidant activity of holy basil extract.

[0114] B. Anti-inflammatory effect The anti-inflammatory activity of holy basil extract was investigated by assaying interleukin 8 (IL8) produced by keratinocytes in response to PMA-induced inflammatory stress.

[0115] A. Materials and Methods Normal human keratinocytes were preincubated for 24 hours in the presence of 0.001%, 0.005% or 0.01% of dried ocitrus extract obtained according to Example 1, or 0.1 μM dexamethasone, an anti-inflammatory reference. The cells were then stimulated by treatment for 24 hours in the presence of 10 μg / mL PMA (phorbol-myristate-acetate).

[0116] At the end of treatment, produced IL8 was quantified by ELISA assay in culture supernatants.

[0117] The results were statistically analyzed using the Student's t test.

[0118] b.Results The results are summarized in Table 9. The oasis extract significantly inhibited PMA-induced IL8 production in keratinocytes, in some cases more significantly than the reference dexamethasone; thus demonstrating anti-inflammatory activity.

[0119] [Table 9]

[0120] C. anti-aging activity Skin ageing is characterized by changes in the dermal matrix, involving a decrease in the quality and quantity of the matrix fibres, primarily collagen and elastin.

[0121] The anti-aging activity of holy basil was assessed by its ability to stimulate the expression and production of dermal matrix markers under standard conditions and under conditions of inflammatory stress that modeled chronic inflammation, also known as "inflammagenesis" or low-grade inflammation.

[0122] C.1. Effects on the dermal matrix The effect of oasis extract on the expression of dermal matrix markers was evaluated in normal human dermal fibroblasts in culture or in reconstructed human skin.

[0123] C.1.1 Stimulation of gene expression of dermal matrix markers in normal human fibroblasts A. Materials and Methods Normal human dermal fibroblasts were incubated for 48 hours in the presence of 0.001% and 0.005% of the dried extract of Holy Basil obtained according to Example 1 or 5 ng / mL of TGFβ1, a reference for a positive assay.

[0124] Gene expression of collagen I, elastin, dermatopontin, and MMP1 (matrix-metalloproteinase 1) was assessed by real-time quantitative PCR.

[0125] Results were statistically analyzed using one-way analysis of variance (ANOVA) followed by Dunnett's post-hoc test.

[0126] b.Results The results are summarized in Table 10. Holy basil extract significantly stimulated the gene expression of collagen I, elastin, and dermatopontin, which are important markers involved in the density of the dermal matrix. Furthermore, holy basil extract significantly inhibited the gene expression of MMP1, an enzyme involved in the degradation of the dermal matrix.

[0127] These results are Densification of the dermal matrix ,therefore anti-aging activity This demonstrates that.

[0128] [Table 10]

[0129] C.1.2 Stimulation of protein expression of dermal matrix markers in normal human fibroblasts A. Materials and Methods Human dermal fibroblasts were treated for 48 hours with 0.001% and 0.005% of the dried extract of Holy Basil obtained according to Example 1, or with a mixture of 10 μM vitamin C and 5 ng / mL TGFβ used as a positive reference.

[0130] Collagen I production was assessed by immunofluorescence labeling and image analysis.

[0131] b.Results The results are summarized in Table 11. Holy basil extract significantly stimulated collagen I labeling in fibroblasts. The results showed that Densification of the dermal matrix This supports the activity of the extract against

[0132] [Table 11]

[0133] C.1.3 Stimulation of protein expression of dermal matrix markers in normal human fibroblasts A. Materials and Methods A preparation (cream) containing 3% basil extract obtained according to Example 1 or its placebo was applied to the surface of reconstructed human skin. After 24 hours of incubation, gene expression of markers of interest for skin effects was evaluated by real-time quantitative PCR.

[0134] b.Results The results are summarized in Table 12. Holy basil extract significantly increased the expression of CLN3 and PXN genes in reconstructed skin.

[0135] CLN3 encodes a lysosomal / endosomal transmembrane protein (ceroid-lipofuscinosis neuron 3) involved in the regulation of the autophagy process. Autophagy is an intracellular detoxification process that allows the degradation and recycling of damaged proteins and organelles. This process is essential to maintain cellular function and homeostasis. Stimulation of autophagy is of interest to delay aging and extend cellular lifespan.

[0136] PXN encodes a focal adhesion molecule (paxillin), whose expression declines with age, making it a target for anti-aging activities. Because paxillin plays a key role in cell-matrix binding and mechanical signaling between fibroblasts and their matrix, its loss with age may impair cellular function and thus initiate or exacerbate skin aging. Inducing its expression may help restore age-related collagen loss and dermal contractility.

[0137] [Table 12]

[0138] C.2. Effects in inflammation-aging models As we age, a subclinical, mild and chronic inflammation can develop in skin tissues. This is the result of cumulative exposure to external stresses throughout life. This inflammation causes tissue damage and the production of reactive oxygen species, resulting in the release of inflammatory cytokines (IL1, IL6, TNF, CRP...) that can accelerate the skin ageing process. This phenomenon of skin ageing associated with inflammatory processes is: inflammation aging It is known as.

[0139] The Ocimum sanctum extract was evaluated in an in vitro model recapitulating the inflammatory process; inflammation, as represented by the stimulation of IL1α production, was induced in reconstituted epidermis. Conditioned medium from these reconstituted epidermis was then applied to fibroblasts and the expression of dermal markers, which are altered by IL1α produced by the epidermis, was evaluated.

[0140] A. Materials and Methods The reconstructed epidermis was treated topically with a preparation (cream) containing 3% Ocimum sanctum extract obtained according to Example 1 or its placebo, or treated globally with 1 μM dexamethasone, an anti-inflammatory reference. After 24 hours of preincubation, the epidermis was stimulated with 0.5 μg / mL PMA and incubated again for 24 hours.

[0141] Culture supernatants were collected and on the one hand IL1α assays were performed and on the other hand these supernatants (conditioned media) were plated onto cultures of normal human dermal fibroblasts.

[0142] Fibroblasts were incubated in the presence of conditioned medium for 24 h, after which expression of markers of interest was assessed by real-time quantitative PCR.

[0143] The results were statistically analyzed using ANOVA followed by Tukey's test.

[0144] b.Results The results are shown in Tables 13 and 14. The 3% formulation of the basil extract significantly inhibited PMA-induced IL1α release on the reconstructed epidermis (Table 13). This effect was significantly higher than that of the placebo.

[0145] Application of conditioned medium from PMA-stimulated reconstituted epidermis to fibroblast cultures induced alterations in gene expression of inflammatory markers, oxidative stress, and dermal matrix synthesis and degradation (Table 14). These results substantiate the inflammation-aging model.

[0146] Under these conditions, the oasis extract modulated the overexpression of cytokines and chemokines (CXCL1, IL8, IL6, CCL2), oxidative stress markers (MT1G, SOD2), matrix degrading enzymes (MMP1, MMP3) and ICAM1. The extract also restored the expression of matrix assembly markers (HAS2, DPT).

[0147] These results indicate that holy basil extract can modulate the inflammatory aging process.

[0148] [Table 13]

[0149] [Table 14]

[0150] D. Defensive activity against various stresses Every day, the skin is exposed to different kinds of exogenous and / or endogenous stresses that can alter its function and quality and ultimately lead to irreversible damage.

[0151] The protective effect of holy basil extract was evaluated in different models representing different external (pollution) or internal (psychological) stresses.

[0152] D.1. Protection against skin damage caused by psychological stress Nitric oxide (NO) is a reactive species involved in the regulation of skin homeostasis. This highly labile molecule is synthesized and released by numerous cell types to regulate physiological and homeostatic responses such as melanogenesis, wound healing, inflammatory regulation, immune regulation, and vasodilation.

[0153] Low levels of NO are important for cell signaling, whereas release of high levels of NO is generally associated with oxidative stress.

[0154] Psychological stress is characterized by activation of the hypothalamic-pituitary-adrenal (HPA) axis, which links the central nervous system and the endocrine system, and an increase in endogenous glucocorticoids such as cortisol.

[0155] Numerous studies have demonstrated that psychological stress negatively affects skin homeostasis, leading to altered immune and inflammatory responses, including the activation of free radicals, including NO.

[0156] Holy basil extract was evaluated in a model reproducing the effects of psychological stress (emotional disorders / anxiety) on the skin by treating keratinocytes with cortisol and quantifying NO as a biomarker of induced stress.

[0157] A. Materials and Methods NO production by normal human skin keratinocytes was measured by continuous amperometry for 10 min after treatment with 1 μM cortisol in the presence of 0.001% and 0.005% dried extract of Holy Basil obtained according to Example 1, or 1 mM ascorbic acid (AA) (positive reference).

[0158] b.Results The results are shown in Figure 1. Holy basil extract significantly inhibited NO production induced by cortisol stress in keratinocytes, thus antagonizing the adverse effects of emotional / psychological stress on skin homeostasis.

[0159] D.2. Protection against DNA damage Air pollutants have adverse effects on many tissues, including the skin. Repeated exposure to pollutants can cause premature aging, depigmentation and DNA damage.

[0160] The protective effect of holy basil extract was evaluated against DNA damage (double strand breaks) induced in keratinocytes by pollutant stress.

[0161] A. Materials and Methods Normal human epidermal keratinocytes were pre-incubated for 24 hours in the presence of 0.001% and 0.005% of the dried extract of Holy Basil obtained in Example 1 or nicotinamide as a reference, before being exposed to camptothecin for 1 hour. Camptothecin is a genotoxic compound and a topoisomerase I inhibitor used to induce DNA double strand breaks.

[0162] Double-strand breaks were detected by immunostaining for γH2AX (phosphorylation of histone H2AX, an early intervening protein in response to double-strand breaks) and quantified by image analysis.

[0163] The results were statistically analyzed using ANOVA followed by Tukey's test.

[0164] b.Results The results are shown in Table 15. The oasis extract significantly inhibited the double-stranded DNA breaks induced by camptothecin treatment. Therefore, this extract Protection against genotoxic pollutant stress Shows activity.

[0165] [Table 15]

[0166] D.3. Protection of mitochondrial function against pollutant stress The protective effect of holy basil extract against urban pollution was investigated in normal human fibroblasts. Cells were exposed to urban pollutants in the presence of the extract and cellular bioenergetics and the occurrence of autophagy / mitophagy were analyzed using BBS+ technology.

[0167] Alterations in mitochondrial function play a central role in the aging process; 90% of age-related ROS production originates from mitochondria.

[0168] Environmental factors (UV radiation, pollution) can impair cellular function and increase oxidative stress, contributing to premature aging.

[0169] Intracellular ROS are primarily formed by dysfunction of the mitochondrial electron transport chain.

[0170] Mitochondrial DNA is particularly sensitive to ROS, a phenomenon that ultimately contributes to the aging process: Decreasing energy production, Increased oxidative stress, Increased mitochondrial function.

[0171] D.3.1 Cellular bioenergetics A. Materials and Methods Normal human skin fibroblasts were incubated for 120 minutes in the presence of 0.1% of the urban pollutant mixture and 0.001% and 0.005% of the dried matter of the Holy Basil extract obtained in Example 1.

[0172] Cellular bioenergetics was assessed using the Bioenergetic Balance Screen Plus (BBS+) method, which includes multiplex quantification of oxygen consumption, ATP production, extracellular lactate levels and the NAD+ / NADH ratio.

[0173] The results were statistically analyzed using Student's t-test.

[0174] b.Results The results are shown in Figure 2. Exposure of cells to urban pollutants induced changes in the respiratory electron transport chain, decreasing oxygen consumption and ATP levels.

[0175] Under these conditions, the holy basil extract dose-dependently increased the oxygen consumption rate and stimulated ATP production. These observations indicate a direct effect of the extract on stimulating and normalizing Krebs cycle function and energy production. The extract is also able to resolve the alterations in the mitochondrial respiratory chain caused by pollutants. Finally, the reduction in lactate production by the extract indicates a reduction in the contribution of de novo glycolysis to ATP production, re-engaging respiratory chain activity in cellular bioenergetics.

[0176] Holy basil extract helps normalize energy balance altered by pollution-induced stress.

[0177] D.3.2 Autophagy / Mitophagy Research A. Materials and Methods Normal human skin fibroblasts were incubated for 120 minutes in the presence of 0.1% of the urban pollutant mixture and 0.001% and 0.005% of the dried matter of the Holy Basil extract obtained in Example 1.

[0178] Analysis of autophagy and mitophagy was assessed via dyes specific for lysosomal markers and damaged mitochondria; damaged mitochondria were visualized by red fluorescence, which is emitted when mitochondrial membrane damage occurs and is generated by the reaction of a fluorescent dye covalently bound to the mitochondrial membrane. Lysosomal dyes were used to label lysosomes and colocalization of damaged mitochondria to lysosomes was assessed.

[0179] Three parameters were analyzed: - percentage of damaged mitochondria per cell; - density of lysosomes per cell (lysosomal index, which represents autophagy); - The percentage of lysosomes containing damaged mitochondria (mitophagy index, which represents mitophagy).

[0180] The results were statistically analyzed using Student's t-test.

[0181] b.Results The results are shown in Figures 3, 4 and 5. When cells were exposed to urban pollutants, the number of damaged mitochondria increased. Holy basil extract potently and significantly suppressed the number of damaged mitochondria (Figure 3).

[0182] During macroautophagy, autophagosomes were formed to transport recycled damaged mitochondrial membranes to the lysosomal compartment. Pollutant stress significantly increased the density of lysosomes per cell, indicating an increase in the autophagy process. Holy basil extract reduced and normalized lysosomal density (Figure 4).

[0183] Mitophagy is expressed as the percentage of lysosomes colocalized with damaged mitochondrial signals. Contamination increased the frequency of lysosomes containing damaged mitochondria, demonstrating an increase in the mitophagy process. Contamination increased mitochondrial and endoplasmic reticulum stress, both of which cause mitochondrial membrane damage. The increase in mitophagy observed under stress is related to the cells' attempt to remove damaged mitochondria and / or malformed proteins or lipids. Holy basil extract reduced and normalized pollutant stress-induced mitophagy (Figure 5).

[0184] Overall, the holy basil extract reversed the effects of pollution on mitochondrial damage and normalized the processes of autophagy and mitophagy. These results indicate a protective activity of the mitochondrial network against external stress.

[0185] E. Healing promotion activities The pro-healing activity of oasis extract was investigated by assessing fibroblast migration in a scratch assay model.

[0186] A. Materials and Methods Mechanical scratches were made on confluent mats of normal human dermal fibroblasts. Immediately after the "scar" was made, the cells were treated with 0.001% and 0.005% dried extract of Holy Basil obtained in Example 1, or 10% dried extract of Holy Basil, a positive control. -5 M insulin was added to the medium and cell migration was monitored over time by immunofluorescence labeling of cell nuclei. The migration rate was calculated.

[0187] b.Results The results are shown in Figure 6. 0.005% holy basil extract stimulated fibroblast migration, indicating the pro-healing activity of the extract.

[0188] F. Moisturizing effect The effect of holy basil extract was evaluated for its ability to induce aquaporin-3 production in keratinocytes.

[0189] Hydration is an important criterion to prevent damage to the skin's barrier function. Aquaporin-3 plays a key role in skin hydration; this transmembrane protein is involved in the transport of water and glycerol into keratinocytes to maintain epidermal hydration.

[0190] A. Materials and Methods Normal human epidermal keratinocytes were incubated for 24 hours in the presence of 0.001% and 0.005% of the dried extract of Holy Basil obtained in Example 1 or retinoic acid as a positive reference. The expression of aquaporin-3 was evaluated by immunostaining and quantified by image analysis.

[0191] b.Results The results are shown in Table 16. 0.001% of the dried extract of Holy Basil significantly increased the expression of aquaporin-3 in keratinocytes, demonstrating the moisturizing activity of the extract.

[0192] [Table 16]

[0193] G. Activity against skin microbiota The effect of holy basil extract was investigated on various bacterial strains representative of the skin microbiome, more specifically on their ability to form biofilms.

[0194] A. Materials and Methods Bacterial strains studied: -Staphylococcus epidermidis MFP04; -Staphylococcus aureus MFP03; - Corynebacterium xerosis CIP100653T / ATCC373; -Micrococcus luteus 0116.

[0195] A1. Study of the effect of extracts on bacterial growth The effect of a 1:50 dilution of the holy basil extract obtained in Example 1 on the growth of six bacterial models was investigated by monitoring the growth for 24 hours (Staphylococcus epidermidis, Staphylococcus aureus) or 48 hours (Micrococcus luteus, Corynebacterium xerosis) under continuous agitation using a multi-well incubator / plate reader. The generation time of each bacterial strain was measured using the wild-type strain as a reference. Statistical differences were determined using the Mann-Whitney test.

[0196] a2. Research into the effects on biofilm formation Biofilm formation of each bacterial species was studied in multi-well plates using crystal violet staining technique. Results were normalized and expressed as a percentage of biofilm formation values ​​in control medium. Statistical differences were determined using the Mann-Whitney test.

[0197] b.Results The results are shown in Tables 17 and 18. Holy basil extract extended the generation time of the pathogenic strain (Staphylococcus aureus) without significantly altering the growth of the symbiotic strain (Table 17).

[0198] Furthermore, holy basil extract strongly and significantly altered the biofilm formation ability of Staphylococcus aureus without altering the biofilm formation ability of the other strains studied (Table 18).

[0199] These results demonstrate the ability of the extract to maintain the balance of the skin microflora.

[0200] [Table 17]

[0201] [Table 18]

[0202] Example 4: Clinical study of the extract of the present invention The clinical anti-aging effect of the holy basil extract of the present invention was evaluated by an in vivo clinical study.For this purpose, a preparation (cream) containing 3% holy basil extract (hereinafter called active) obtained according to Example 1, or its placebo, was evaluated.

[0203] A. Protocol The study was a double-blind, placebo-controlled study. Subjects applied a cream containing holy basil extract (active substance) or placebo (randomized at inclusion) twice daily (morning and evening) for 56 days.

[0204] The composition of the placebo is as follows: [Table 19]

[0205] Inclusion criteria defined for recruited subjects were as follows: ·Gender: Female Age: 40~60 years old Photo type: I~III Race: White Subjects with wrinkles (crow's feet, forehead) Do not use cosmetics on days 6-0

[0206] Efficacy was assessed using the following procedures (measured at D0, D28 and D56): Image Analysis - Topography Measurement of biomechanical skin properties Cross-polarized photography

[0207] B. Evaluation of anti-aging effects on wrinkles a. Results for crow's feet The results obtained by topographical analysis at D0, D28 and D56 are shown in the table below.

[0208] The active ingredient significantly reduced the area and length of the wrinkles detected at D56, showing a significant difference compared to placebo.

[0209] [Table 20]

[0210] b. Effect on forehead wrinkles The results obtained by topographical analysis at D0, D28 and D56 are shown in the table below.

[0211] Regarding the overall area: the active ingredient significantly improved overall forehead wrinkles at D28, but no change at D56. The placebo caused no significant change at D28, but an overall significant worsening at D56. Overall, significant differences in favor of the active ingredient were found at D28 and D56 for all anatomical parameters evaluated.

[0212] [Table 21]

[0213] d. Conclusion – Anti-wrinkle effect These results showed anti-wrinkle effects significantly different from placebo on crow's feet (improvement in wrinkle area and length) and forehead wrinkles (improvement in anatomical parameters).

[0214] C. Evaluation of anti-aging effects on skin biomechanical properties The results obtained by Cutometry at D0, D28 and D56 are shown in the table below. [Table 22]

[0215] The active ingredient improved elasticity (Ur and Ua) at D56, whereas the placebo caused a significant deterioration. The difference between the active and the placebo was significant in favor of the active for both parameters.

[0216] The active ingredient improved Ur / Uf total elasticity at D56, the difference was significant when compared to placebo, which caused a deterioration.

[0217] The active ingredient significantly improved Ua / Uf biological elasticity at D28 and D56. At D56, the difference was significant when compared with placebo, which caused a deterioration.

[0218] The active ingredient improved the net elasticity Ur / Ue at D56. The difference was significant when compared with placebo, which caused a deterioration at D56.

[0219] The active ingredient improved Q1 viscoelastic recovery at D28 and significantly improved it at D56, the difference being more pronounced when compared with placebo, which caused a deterioration at D56.

[0220] The active ingredient improved Q2 elastic resilience at D56, the difference being significant when compared with placebo, which caused a deterioration at D56.

[0221] The active ingredient significantly improved Q3 viscosity recovery on D28 and D56. On D56, the difference was significant when compared to placebo, which showed no significant change.

[0222] Conclusion – Biomechanical properties: The active ingredient significantly improved the biomechanical properties of the skin compared to placebo, improving skin turgor and elasticity.

[0223] D. Evaluation of anti-aging effects on skin tone: color and uniformity measured by photography The results obtained by image analysis at D0, D28 and D56 are shown in the table below. [Table 23]

[0224] The active ingredient is D28 and D56 for redness a * This difference favours the active ingredient on D28 and D56 compared to placebo, which also showed a significant reduction in redness on D28 and D56.

[0225] The active ingredient reduced hemoglobin at D28 and D56.

[0226] The active ingredient significantly improved skin tone evenness from H76 to D56.

[0227] Conclusion – Skin Color Evaluation The active ingredient significantly improved skin brightness, redness and evenness.

[0228] E. Summary of Clinical Trial Results · Anti-aging effect against wrinkles: The active ingredient was instrumentally monitored for anti-aging effects on upper facial wrinkles, i.e. crow's feet and forehead wrinkles. Significant differences were observed over time in favor of the active ingredient compared to placebo.

[0229] Anti-aging effects on biomechanical properties: The active ingredient significantly improved skin turgor and elasticity at D28 and D56, with significant differences in favour of the active ingredient compared to placebo, depending on the parameter studied.

[0230] Skin tone and uniformity impact: The active ingredient improved skin tone evenness on D56, with the difference favoring the active ingredient compared to placebo.

Claims

1. A polyphenol-rich extract of the aerial parts of holy basil, Ocimum sanctum, comprising at least 6% by weight of polyphenols based on the total weight of the dry extract.

2. 2. The extract according to claim 1, wherein the extract comprises at least 5% by weight of hydroxycinnamic derivatives based on the total weight of the dry extract.

3. 2. The extract according to claim 1, wherein the extract comprises at least 2.5% by weight of flavonoids, expressed as rutin equivalents, relative to the total weight of the dry extract.

4. The extract described in claim 1, wherein the extract is obtained by solid / liquid extraction of the aerial parts of the holy basil in a solvent selected from a binary mixture of water / glycol or water / glycerol.

5. The extract described in claim 4, wherein the extract is obtained by solid / liquid extraction of the aerial parts of the holy basil in a solvent selected from a binary mixture of water / glycol or water / glycerol, wherein the proportion of glycol or glycerol in water is between 30% and 90%.

6. A method for preparing a polyphenol-rich extract of the aerial parts of Holy Basil according to any one of claims 1 to 3, said method comprising at least one step of solid / liquid extraction in a solvent selected from the binary mixtures water / glycol or water / glycerol.

7. The method of claim 6, wherein the solvent is selected from a binary mixture of water / glycol or water / glycerol, the proportion of glycol in water or glycerol in water being between 30% and 90%.

8. The method of claim 7, further comprising the steps of: a) grinding the dried aerial parts of holy basil; b) extracting said aerial parts in a solvent selected from a binary mixture of water / glycol or water / glycerol; c) separating the solid phase from the liquid phase by decantation, centrifugation, or continuous filtration; and d) Optionally, drying the extract obtained in step c). The method of claim 6, comprising:

9. The method of claim 6, wherein the method further comprises a step of vacuum evaporation or deodorization.

10. The method of claim 6, wherein the glycol is propanediol or propylene glycol.

11. A composition comprising an extract of the aerial parts of Holy Basil according to any one of claims 1 to 5 or an extract obtained by the method according to any one of claims 6 to 10 as an active ingredient, and also comprising an excipient.

12. - disorders or lesions of the skin, mucous membranes or skin appendages; - Vascular disorders; or - Microbiome imbalance An extract according to any one of claims 1 to 5, or an extract obtainable by a method according to any one of claims 6 to 10, or a composition according to claim 11, for use in the prevention or treatment of

13. - the skin, mucosa or skin appendage disorder or lesion is selected from sensitive skin, reactive skin, reactive mucosa, rough skin, inflamed skin, barrier or homeostasis disorders, photosensitized skin, mechanical or thermal insults; - the vascular disorder is selected from redness and couperose; and the imbalance of the microbiota is selected from atopic dermatitis, eczema, occurrence of unpleasant axillary odor, weakening of the skin barrier, acne, psoriasis, sweat gland abscess, folliculitis, neonatal scalp dermatitis, dandruff, pruritus, itching, candidiasis and bacterial vaginosis, 13. An extract or composition for use as claimed in claim 12.

14. A method for the cosmetic care of the skin, the skin appendages or the mucous membranes, aimed at improving their condition or appearance, which comprises administering an extract according to any one of claims 1 to 5, or an extract obtainable by a method according to any one of claims 6 to 10, or a composition according to claim 11.

15. - For the purpose of moisturizing the skin or mucous membranes, or preventing damage to the skin barrier and its dehydration, or preventing skin aging, - for the purpose of improving the firmness, elasticity or length of the skin or enhancing the mechanical properties of the skin and mucous membranes, or - To improve the evenness of skin tone or reduce skin blemishes, 15. A cosmetic care method according to claim 14.

16. A cosmetic care method as described in claim 15 for the purpose of preventing dry, shriveled, bloated or saggy skin, or for enhancing or restoring skin elasticity or firmness, or for the treatment of fine lines and wrinkles.

Images