Camellia oleifera tree oil concentrate and application thereof

Abstract

The present invention relates to a lipid extract of camellia oleifera oil concentrated into an unsaponifiable fraction, a cosmetic, pharmaceutical or dermatological composition comprising suitable excipients and said lipid extract, and uses thereof.

Description

Technical Field

[0001] The present invention relates to a lipid extract of Camellia oleifera tea oil concentrated into an unsaponifiable fraction, and a cosmetic composition, a nutritional composition, a pharmaceutical composition or a dermatological composition comprising a suitable excipient and the lipid extract.

[0002] Another object of the present invention is a method for preparing a lipid extract of Camellia oleifera oil concentrated into an unsaponifiable fraction. The present invention also relates to the use of the composition or extract for preventing or treating diseases or conditions of the skin, mucous membranes, or appendages. Finally, the present invention relates to a method for cosmetic treatment of the skin, appendages, or mucous membranes to improve their condition or appearance, comprising applying the composition or extract to improve, in particular, the user's physical / physiological and psychological well-being. Background Art

[0003] Camellia (Camellia) originates in East Asia and includes more than 200 kinds of evergreen woody plant species.Some species have significant economic value, such as tea tree (C.sinensis), camellia (C.japonica) and oil tea tree (C.oleifera). Tea tree is mainly planted in tropical and subtropical regions, for making tea, while camellia is planted in temperate regions, as an ornamental plant, and its oil is commonly used as a cosmetic in Japan. Oil tea tree (also referred to as oil-tea camelia) can be grown on poor land without fertilizer, starts to bear fruit after initial planting six years, and keeps high yield for 80 years. This species is mainly planted in China, for producing edible oil (" camellia oil", "tea seed oil") (Yang et al., 2016).

[0004] The amount of oil and fatty acid composition in seeds varies depending on species, soil, and climate (Hu & Yang, 2018). Wen et al. (2018) estimated that light intensity during fruit development and ripening (July to October) affects fruit yield and quality.

[0005] Regarding species diversity, Yang et al. (2016) compared seeds from 10 different varieties and several wild individuals and concluded that the oil content and fatty acid profiles of their extracts were not affected by artificial cultivation and variety selection.

[0006] Tea tree oil is used in cooking. It's also commonly used to treat stomach aches and burns. Because its monounsaturated fatty acid content (primarily C18:1) is similar to olive oil, several studies have been conducted to determine whether it offers the same health benefits, particularly in reducing cardiovascular risk. Studies have shown that tea oil can lower cholesterol, triglyceride, and low-density lipoprotein (LDL) levels and reduce thromboxane B2 and lipid peroxidation products. It also increases the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Some of this activity may be attributed to the presence of saponins.

[0007] The applicant has discovered that a lipid extract of Camellia oleifera oil concentrated into an unsaponifiable fraction has hitherto undescribed cosmetic and dermatological properties.

[0008] In particular, this is the first time that such lipid extracts have been used in this way for their specific properties. Thus, surprisingly, the applicant has discovered that a lipid extract of Camellia oleifera oil concentrated into an unsaponifiable fraction can be used to stimulate, restore or regulate the metabolism of skin and mucosal cells and / or for the prevention and / or treatment of diseases associated with skin tissue. Summary of the Invention

[0009] According to a first aspect, the present invention relates to a lipid extract of tea oil, which is tea tree oil concentrated into an unsaponifiable fraction, containing 1.5% to 100% by weight, preferably 1.8% to 80% by weight, and preferably 2% to 60% by weight of unsaponifiable matter relative to the total weight of the extract.

[0010] According to a second aspect, the present invention relates to a method for preparing said extract, comprising the following consecutive steps:

[0011] a) molecular distilling the crude oil or refined oil of Camellia oleifera;

[0012] b) extraction of unsaponifiable matter, where applicable;

[0013] c) recovering the concentrated unsaponifiable oil obtained after step a) or the unsaponifiable matter obtained after step b);

[0014] d) optionally carrying out a step of deodorizing and / or decolorizing the concentrated unsaponifiable oil or the unsaponifiable matter.

[0015] Advantageously, step b) comprises the following consecutive steps:

[0016] i. after step a) the tea oil concentrated into the unsaponifiable fraction obtained is saponified,

[0017] ii. The unsaponifiable matter is then extracted using a suitable solvent such as ethyl acetate and hexane.

[0018] According to a third aspect, the present invention relates to a composition comprising the lipid extract of tea oil and a suitable excipient, wherein the lipid extract is an active ingredient. The composition may be a cosmetic composition, a nutritional composition, a pharmaceutical composition or a dermatological composition.

[0019] According to a fourth aspect, the present invention relates to an extract according to the invention or a composition comprising an extract according to the invention or an extract obtainable by a process according to the invention, for use in a dermatological or pharmaceutical composition.

[0020] According to a fifth aspect, the present invention relates to the use of an extract according to the invention or a composition comprising an extract according to the invention or an extract obtainable by a process according to the invention in a nutraceutical composition or as a food supplement.

[0021] According to a sixth aspect, the present invention relates to a method for the cosmetic care of the skin and / or appendages and / or mucous membranes in order to improve their condition and / or appearance, comprising the topical or oral administration of an extract according to the invention or a composition according to the invention.

[0022] According to a seventh aspect, the present invention relates to a method for the cosmetic treatment of dry skin, xerosis, sensitive skin, irritated skin, comprising the topical or oral administration of an extract according to the invention or a composition according to the invention.

[0023] Detailed description of the invention

[0024] The present invention relates to a lipid extract of tea oil, which is tea oil from the oleifera Camellia sinensis plant concentrated into an unsaponifiable fraction containing from 1.5% to 100% by weight, preferably from 1.8% to 80% by weight, preferably from 2% to 60% by weight of unsaponifiable matter relative to the total weight of the extract.

[0025] Unsaponifiable matter is the fraction of fatty matter that remains insoluble in water after prolonged exposure to alkali and can be extracted with organic solvents. Five major groups of substances are present in the majority of the unsaponifiable matter of vegetable oils: saturated or unsaturated hydrocarbons, aliphatic or terpene alcohols, sterols, tocopherols, tocotrienols, carotenoid pigments and xanthophylls.

[0026] The oil is concentrated into an unsaponifiable fraction comprising, in particular, greater than 1.5% by weight of unsaponifiable matter, advantageously greater than 2% by weight and preferably greater than 2.5% by weight of unsaponifiable matter relative to the total weight of the extract.

[0027] The lipid extract of the tea tree camellia oil that is condensed into the unsaponifiable matter fraction advantageously comprises the fatty acid with 12 to 22 carbon atoms, more advantageously 16 to 20 carbon atoms.These fatty acids can be saturated, monounsaturated or polyunsaturated.This concentrate advantageously is available from refined tea oil.Oil can be refined according to method well known to persons skilled in the art, for example physical refining (degumming with water, by stripping deodorization deacidification) and chemical refining (degumming by water or acid treatment to remove phosphatide, use alkaline solution and free fatty acids, decolouring, refrigeration and deodorization), particularly by stripping deodorization.The feature example of the tea tree camellia oil of refining is provided in the following table.

[0028] Table 1

[0029]

[0030] Starting from this oil, the Camellia oleifera oil is advantageously concentrated into an unsaponifiable fraction - a "concentrate", usually by molecular distillation. The oil concentrated into the unsaponifiable fraction advantageously has the following detailed description.

[0031] Table 2

[0032]

[0033]

[0034] Advantageously, the unsaponifiable matter fraction is as described below.

[0035] Table 3

[0036]

[0037] Relative percentage: relative to the total weight of sterols

[0038] According to a preferred embodiment of the present invention, the α-tocopherol content is greater than 60% by weight, preferably 80% by weight, preferably 95% by weight relative to the total weight of tocopherol.

[0039] According to another preferred embodiment, the sterol content of the extract according to the invention is greater than 1.5% by weight, preferably 2% by weight, relative to the total weight of the extract.

[0040] The lipid extract according to the invention advantageously contains preferably 10% to 50% by weight, preferably 15% to 40% by weight, more preferably 15% to 35% by weight, typically 20% to 35% by weight of β-amyrin and Δ5-avenasterol, relative to the total weight of sterols, and / or preferably 10% to 50% by weight, preferably 15% to 40% by weight, more preferably 15% to 35% by weight, typically 20% to 35% by weight of lupeol and Δ7-stigmasterol, relative to the total weight of sterols.

[0041] Advantageously, the lipid extract according to the invention comprises:

[0042] Table 4

[0043]

[0044]

[0045] The extract according to the invention is advantageously obtained by a process comprising the following successive steps:

[0046] a) molecular distilling the crude oil or refined oil of Camellia oleifera;

[0047] b) extraction of unsaponifiable matter, where applicable;

[0048] c) recovering the concentrated unsaponifiable oil obtained after step a) or the unsaponifiable matter obtained after step b); and

[0049] d) optionally carrying out a step of deodorizing and / or decolorizing the concentrated unsaponifiable oil or the unsaponifiable matter.

[0050] When an oil enriched in (concentrated in) unsaponifiable matter is to be recovered, step b) is not carried out. When an unsaponifiable matter fraction is to be recovered, step b) is carried out.

[0051] This step b) advantageously comprises the following successive steps:

[0052] i. after step a) the tea oil concentrated into the unsaponifiable fraction obtained is saponified,

[0053] ii. The unsaponifiable matter is then extracted using a suitable solvent such as ethyl acetate and hexane.

[0054] The present invention also relates to a process for preparing an extract according to the invention comprising the following successive steps:

[0055] a) molecular distilling the crude oil or refined oil of Camellia oleifera;

[0056] b) extraction of unsaponifiable matter, where applicable;

[0057] c) recovering the concentrated unsaponifiable oil obtained after step a) or the unsaponifiable matter obtained after step b);

[0058] d) optionally carrying out a step of deodorizing and / or decolorizing the concentrated unsaponifiable oil or the unsaponifiable matter.

[0059] The molecular distillation step a) is preferably carried out using an apparatus selected from a centrifugal molecular still and a wiped film molecular still.

[0060] Centrifugal molecular stills are well known to those skilled in the art. For example, application EP 493 144 has described this type of molecular still. Usually, the product to be distilled is applied to the heated surface (hot surface) of a conical rotor rotating at high speed in the form of a thin layer. The still is placed under vacuum. Under these conditions, the composition of the oil, such as unsaponifiable matter, evaporates from the hot surface rather than boiling, and the advantage is that oil and its composition, particularly unsaponifiable matter (these products are considered to be vulnerable), can not degrade in the evaporation process.

[0061] Wiped film molecular stills are also known to those skilled in the art. Typically, they include a still chamber equipped with a rotating scraper that allows continuous application of the product to be distilled on the evaporation surface (hot surface). The product vapor is condensed by a freezing finger placed in the center of the still chamber. The peripheral feed and vacuum systems are very similar to those of centrifugal stills (feed pumps, blades, and oil diffusion vacuum pumps, etc.). The recovery of residue and distillate is accomplished by gravity flow into a suitable container.

[0062] At the end of the fractionation step, the distillation fraction rich in unsaponifiables advantageously represents from 3% to 15% by weight of the initial oil and the distillation fraction rich in triglycerides advantageously represents from 85% to 97% by weight of the initial oil.

[0063] Furthermore, it was confirmed that this method does not cause any chemical modification or alteration of the unsaponifiable matter compounds and retains the highly unsaturated fraction. As a result, the fatty acid distribution of the concentrated tea oil is the same as that of the tea oil before concentration.

[0064] After this molecular distillation step, the product obtained can optionally be deodorized and / or decolorized by methods known to those skilled in the art, for example by stripping or molecular distillation, or by treatment on activated carbon.

[0065] The tea oil thus obtained, concentrated into the unsaponifiable fraction, has the above-mentioned characteristics.

[0066] When it is desired to recover an oil rich in unsaponifiable matter, step b) is not carried out.

[0067] When the unsaponifiable matter fraction is to be recovered, step b) is carried out. This step advantageously comprises the following successive steps:

[0068] i. after step a) the tea oil concentrated into the unsaponifiable fraction obtained is saponified,

[0069] ii. The unsaponifiable matter is then extracted using a suitable solvent such as ethyl acetate and hexane.

[0070] The unsaponifiable matter of tea oil can be obtained by methods known to those skilled in the art. For example, it can be obtained by saponifying tea oil concentrated into an unsaponifiable matter fraction and then extracting the unsaponifiable matter using a suitable solvent. Typically, the unsaponifiable matter is then washed until the saponifiable matter is completely removed, and the solvent is then evaporated. Finally, the unsaponifiable matter is advantageously deodorized with steam and then stripped with nitrogen to eliminate traces of solvent.

[0071] The unsaponifiable matter of tea oil thus obtained advantageously has the characteristics described above.

[0072] In the remainder of the description, "an extract according to the invention" will be understood to mean the extract as defined above per se or an extract obtainable by a process according to the invention as described above.

[0073] Another object of the present invention relates to a composition comprising the lipid extract of Camellia oleifera oil according to the present invention as an active ingredient.

[0074] The composition according to the invention advantageously comprises from 0.01% to 20% by weight, advantageously from 0.05% to 15% by weight and more advantageously from 0.1% to 10% by weight of said lipid extract relative to the total weight of the composition.

[0075] The lipid extract according to the invention is advantageously used as active agent in a composition such as a cosmetic, nutraceutical, dermatological or pharmaceutical composition, which may comprise one or more suitable excipients.

[0076] The composition according to the present invention can be formulated in the form of different preparations suitable for topical or oral administration.

[0077] Alternatively, various formulations are suitable for topical administration and include creams, emulsions, lotions, ointments, lotions, oils, aqueous or hydroalcoholic or glycolic solutions, powders, patches, sprays, shampoos, lacquers, or any other product for external use.

[0078] In the case of cosmetic, pharmaceutical or dermatological use, the composition is advantageously formulated in the form of a preparation suitable for topical administration.

[0079] In the case of nutritional supplement use, the composition is advantageously formulated in the form of a preparation suitable for oral administration. The extract according to the present invention may be included in a food supplement or a nutritional composition. The food supplement may be in the form of the extract according to the present invention itself, or in the form of a gelatin or vegetable capsule or softgel. The food supplement may then contain 10% to 100% by weight of the extract according to the present invention.

[0080] The composition of the present invention can be directly incorporated into nutritional products, dietary products (especially high protein products) or beverages using techniques such as mixing, pouring, injecting, compounding, absorbing, kneading and spraying without further modification.

[0081] Composition according to the present invention can also contain adjuvant commonly used in medicine, dermatology, beauty or nutrition product field, for example hydrophilic or lipophilic gelling agent, hydrophilic or lipophilic active ingredient, thickening agent, preservative, antioxidant, solvent, spices, chelating agent, odor absorbent, chemical or mineral filter, mineral pigment, surfactant, polymer, silicone oil and coloring matter.The amount of these different adjuvants is the amount commonly used in the field under consideration, for example, is 0.01% to 20% of the composition total weight.These adjuvants can be incorporated in fat phase, aqueous phase, lipid vesicle body and / or nanoparticle according to their property.Those skilled in the art can select suitable adjuvant according to expected application.

[0082] As oils that can be used in the compositions for implementing the invention, mention may be made of mineral oils, oils of vegetable origin (almond oil, sunflower oil, plum oil), oils of animal origin, synthetic oils, silicone oils and fluorinated oils (perfluoropolyethers). Fatty alcohols (cetyl alcohol), fatty acids and waxes (beeswax) may also be used as fats.

[0083] As emulsifiers and co-emulsifiers that can be used in the present invention, for example, esters of fatty acids with polyethylene glycols such as PEG-40 stearate and PEG-100 stearate, and esters of fatty acids with polyols such as glyceryl stearate and sorbitan tristearate may be mentioned.

[0084] As hydrophilic gelling agents, mention may in particular be made of carboxyvinyl polymers (carbomers), acrylic acid copolymers such as acrylate / alkyl acrylate copolymers, polyacrylamides, polysaccharides, natural gums and clays, while as lipophilic gelling agents, mention may be made of modified clays such as bentonite, metal salts of fatty acids, hydrophobic silica and polyethylene.

[0085] The mode of administration, dosage and optimal galenic form of the composition according to the invention can be determined according to the criteria usually taken into account when establishing a medical, dermatological or cosmetic treatment suitable for a patient, such as the extent of the skin area to be treated, tolerance to treatment, skin type.

[0086] The invention also relates to the use of an extract according to the invention or a composition according to the invention as a food supplement or in a nutraceutical composition.

[0087] The invention also relates to an extract according to the invention or a composition according to the invention for use as or in a dermatological or pharmaceutical composition.

[0088] The compositions or extracts according to the present invention can be used to stimulate, restore or regulate the metabolism of skin and mucosal cells and / or prevent and / or treat diseases associated with skin tissue. Advantageously, the compositions or extracts according to the present invention are used to prevent and / or treat diseases or conditions of the skin and / or mucosa and / or appendages.

[0089] Preferably, the diseases or conditions of the skin and / or mucous membranes and / or appendages are selected from inflammatory reactions, diseases associated with endogenous or exogenous stress (e.g. psychological stress, stress associated with anxiety states, stress associated with free radical attack with or without chemical or atmospheric pollution and / or exposure to UV or IR), disorders of the barrier or homeostasis, photosensitive skin, mechanical and / or thermal insults, or as healing agents. Diseases or conditions of the skin and / or mucous membranes and / or appendages can also be selected from diseases or conditions caused by an imbalance of the microbiota; they are advantageously selected from atopic dermatitis, eczema, the development of unpleasant axillary odor, a weakening of the skin barrier, acne, psoriasis, hidradenitis suppurativa, folliculitis, cradle cap, dandruff, itching, in particular itching of the scalp, irritation, candidiasis and bacterial vaginosis.

[0090] Advantageously, the inflammation, irritant reaction, disease or condition or skin barrier disorder or homeostasis disorder is: acne, rosacea or lupus erythematosus, vascular diseases, in particular redness and redness, diaper dermatitis, atopic dermatitis, eczema, contact dermatitis, irritant dermatitis, allergic dermatitis, seborrheic dermatitis (cradle cap), sensitive skin, reactive skin, dry skin (xerosis), dehydrated skin, skin redness, skin erythema, aged or photoaged skin, photosensitive skin, pigmented skin (melasma, post-inflammatory hyperpigmentation, etc.), skin with stretch marks, sunburn, irritation caused by chemical agents or physical factors (e.g. stress in pregnant women), bacteria, fungi, skin aging, in particular photoaging and diseases associated with free radical attack involving chemical or atmospheric pollution and / or involving exposure to UV or IR.

[0091] Particularly advantageously, the disease or condition of the skin and / or mucous membranes and / or appendages is selected from the group consisting of dry skin, xerosis, sensitive skin and irritation.

[0092] Advantageously, the present invention also relates to an extract according to the invention or a composition according to the invention for use in preventing and / or treating dry skin, xerosis, sensitive skin, irritation, delicate lips, cheilitis, atopic dermatitis, eczema and psoriasis. These diseases may be skin diseases associated with endogenous or exogenous stress, such as psychological stress.

[0093] The composition or extract according to the invention is advantageously able to improve skin comfort.

[0094] According to a specific embodiment, the extract or composition according to the invention can be used as an anti-aging and / or healing agent; for preventing skin damage and / or maintaining homeostasis of the skin or mucous membranes or as an antioxidant and / or anti-inflammatory agent. Advantageously, the composition or extract according to the invention can be used to prevent or delay premature skin aging, in particular photo-induced skin aging, and advantageously for preventing, reducing and / or treating wrinkles, fine lines, or micro-undulations.

[0095] The composition or extract according to the invention may also be used to promote healing.

[0096] According to a particular technical feature, the composition or extract according to the invention can also be used for preventing and / or treating conditions or diseases chosen from epidermal scars, delicate lips and cheilitis, stretch marks, skin after bites, skin abrasions, spots and / or skin scabs, and delicate and sensitive skin, advantageously chosen from delicate and sensitive skin.

[0097] The present invention also relates to the use of an extract according to the invention or a composition according to the invention for the preparation of a cosmetic, pharmaceutical or dermatological composition for the prevention and / or treatment of diseases or conditions of the skin and / or mucous membranes and / or appendages. Preferably, the diseases or conditions of the skin and / or mucous membranes and / or appendages are selected from inflammatory reactions, diseases associated with endogenous or exogenous stress (e.g. psychological stress, stress associated with anxiety states, stress associated with free radical attacks with or without chemical or atmospheric pollution and / or exposure to UV or IR), disorders of barrier or homeostasis, photosensitive skin, mechanical and / or thermal aggression.

[0098] The present invention further relates to a method for preventing and / or treating diseases or conditions of the skin and / or mucous membranes and / or appendages, comprising administering, in particular topically, to a subject in need thereof an effective amount of an extract according to the invention or a composition according to the invention. Preferably, the disease or condition of the skin and / or mucous membranes and / or appendages is as defined above.

[0099] Another object of the invention relates to a non-therapeutic cosmetic care method for the skin and / or appendages and / or mucous membranes to improve their condition and / or appearance, advantageously to improve the firmness, elasticity or firmness of the skin, comprising the topical or oral administration of an extract or composition according to the invention.

[0100] The cosmetic care method is preferably carried out to improve skin comfort, generally on healthy skin, healthy appendages or healthy mucous membranes, and is particularly advantageous for the treatment and / or prevention of dry skin, xerosis, sensitive skin and irritated skin.

[0101] The cosmetic use and cosmetic care method according to the invention are generally carried out on healthy skin or healthy parts of the body and are non-therapeutic.

[0102] Another object of the present invention relates to a method for the cosmetic treatment of dry skin, xerosis, sensitive skin, irritated skin, comprising the topical or oral administration of an extract or composition according to the invention.

[0103] Dry skin, xerosis, sensitive and irritated skin can lead to a feeling of tightness and a lack of comfort and softness. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] Figure 1 Active versus placebo results for each of the ten parameters in the "Sensitivity Scale" (left: active, right: placebo). Significance of D0 versus D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of active versus placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1).

[0105] Figure 2 = Overall active ingredient vs. placebo score in the "Sensitivity Scale" (left: active ingredient, right: placebo). Significance of D0 vs. D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of active ingredient vs. placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1).

[0106] Figure 3 : Changes in acoustic stress response (left: active ingredient, right: placebo). Significance of D0 versus D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of active ingredient versus placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1).

[0107] Figure 4 : Changes in cardiac coherence (left: active ingredient, right: placebo). Significance of D0 versus D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of active ingredient versus placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1).

[0108] Figure 5: Changes in galvanic skin response (left: active ingredient, right: placebo). Significance of D0 versus D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of active ingredient versus placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1).

[0109] Figure 6 : Evaluation of the skin moisture test (left: active ingredient, right: placebo) and hydration (left: active ingredient, right: placebo). Significance of D0 versus D28: * = significant (α < 0.05), ° = borderline significant (0.05 < α < 0.1). Significance of the active ingredient versus placebo: solid line = significant (α < 0.05), hatched line = borderline significant (0.05 < α < 0.1). DETAILED DESCRIPTION

[0110] Example

[0111] Example 1: Concentrate according to the invention

[0112] An example of a concentrate according to the invention is a camellia oleifera refined oil concentrated into an unsaponifiable fraction. The extract is prepared by molecular distillation.

[0113] Vacuum conditions: 10 -2 to 10 -3 millibar

[0114] The distillate temperature is between 250℃ and 280℃

[0115] The mass distribution is shown in the table below.

[0116] Table 5

[0117]

[0118] The mass distribution of tocopherol is shown in the table below.

[0119] Table 6

[0120] Percentage of α-tocopherol 100 Percentage of beta-tocopherol 0 Percentage of delta-tocopherol 0 Percentage of gamma-tocopherol 0

[0121] The mass distribution of sterols is shown in the table below.

[0122] Table 7

[0123]

[0124] Example 2: In vitro biological activity test of the extract according to the present invention

[0125] The extract tested was the concentrate from Example 1. It was referred to in the study as "tea oil concentrate." Unless otherwise stated, percentages are expressed as the weight of the concentrate relative to the total weight of the composition tested.

[0126] Anti-inflammatory activity

[0127] The anti-inflammatory activity of tea oil concentrate (THE) was investigated by measuring interleukin-1α (IL-1α) production by keratinocytes in response to PMA (phorbol myristate acetate)-induced pro-inflammatory stress in normal human keratinocyte models, reconstructed epidermis models subjected to PMA (phorbol myristate acetate) stress, and reconstructed epidermis models subjected to SDS (sodium dodecyl sulfate) stress.

[0128] Effects in normal human keratinocyte models

[0129] Materials and methods

[0130] Normal human keratinocytes were pre-incubated for 24 hours in the presence of 0.001%, 0.005%, or 0.01% tea tree oil concentrate or 0.1 μM dexamethasone (anti-inflammatory reference) and then stimulated by overnight treatment with 10 μg / mL PMA (phorbol myristate acetate).

[0131] At the end of the treatment, IL-1α produced in the culture supernatants was quantified by ELISA assay.

[0132] The results were statistically analyzed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test.

[0133] result

[0134] Tea oil concentrate significantly inhibited PMA-induced IL-1α production in keratinocytes; therefore, it has anti-inflammatory potential.

[0135] Table 8: Amount of interleukin-1α produced by keratinocytes subjected to PMA-induced inflammatory stress

[0136]

[0137] Effects in a reconstructed epidermis model subjected to PMA stress

[0138] Materials and methods

[0139] The reconstructed epidermis was topically treated with a formulation (cream) containing 1% tea tree oil concentrate or a placebo, or systemically treated with 1 μM dexamethasone (anti-inflammatory reference). After a 24-hour preincubation, the epidermis was stimulated with 0.5 μg / mL PMA and incubated again for 24 hours.

[0140] The culture supernatant was collected and then subjected to IL-1α assay.

[0141] The results were statistically analyzed using ANOVA followed by Tukey's test.

[0142] result

[0143] Tea oil concentrate formulated at 1% significantly inhibited the release of IL-1α from PMA-induced reconstructed epidermis, an effect that was significantly different from that of placebo.

[0144] Table 9: Amount of IL-1α produced by reconstructed epidermis stimulated with PMA

[0145]

[0146] Effects in a reconstructed epidermis model subjected to SDS stress

[0147] Materials and methods

[0148] The reconstructed epidermis was topically treated with a formulation (cream) containing 1% tea tree oil concentrate or placebo. After a 24-hour pre-incubation, the epidermis was stimulated with 0.1% SDS for 30 minutes, then treated with the product for another 24 hours and incubated for another 24 hours.

[0149] The culture supernatants were collected and then IL-1α and IL8 were measured by ELISA assay.

[0150] The results were statistically analyzed using ANOVA followed by Dunnett's test.

[0151] result

[0152] Tea tree oil concentrate formulated at 1% inhibited SDS-induced IL-1α and IL-8 release.

[0153] Table 10: Amount of IL-1α produced by reconstructed epidermis stimulated with PMA

[0154]

[0155] These diverse results demonstrate the anti-inflammatory potential of tea oil concentrate.

[0156] Anti-aging activity

[0157] Skin aging is characterized by alterations in the dermal matrix, including a decrease in the quality and quantity of matrix fibers.

[0158] The anti-aging activity of tea oil concentrate was evaluated by its ability to stimulate the expression and production of dermal matrix markers under standard conditions and under inflammatory stress conditions simulating "inflammaging."

[0159] Stimulation of gene expression of dermal matrix markers in normal human fibroblasts

[0160] The effect of tea oil concentrate on the expression of dermal matrix markers in cultured normal human dermal fibroblasts was evaluated.

[0161] Materials and methods

[0162] Normal human dermal fibroblasts were incubated for 48 hours in the presence of 0.001% and 0.005% of tea oil concentrate or 5 ng / mL of TGFβ1 (positive reference of the test).

[0163] Collagen III and fibrillin 1 gene expression was evaluated by quantitative real-time PCR.

[0164] The results were statistically analyzed by one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test.

[0165] a. result

[0166] Tea oil concentrate significantly stimulated the gene expression of collagen III and fibrillin 1.

[0167] These results suggest a densification of the dermal matrix due to skin anti-aging effects.

[0168] Table 11: Gene expression of dermal matrix markers in fibroblasts

[0169]

[0170] Role in the "Inflammatory Aging" Model

[0171] As we age, low-grade, asymptomatic, but chronic inflammation may develop within the skin. This is the result of cumulative exposure to external stressors throughout life. This inflammation leads to the release of inflammatory cytokines (such as IL1, IL6, TNF, and CRP) (which can cause tissue damage) and the production of reactive oxygen species, accelerating the skin aging process. This skin aging phenomenon associated with inflammatory processes is known as "inflammatory aging."

[0172] Tea oil concentrate was evaluated in an in vitro model recapitulating the "inflammatory aging" process: inflammation, characterized by stimulation of IL-1α production, was induced in reconstructed epidermis. Conditioned culture medium from these reconstructed epidermis was then applied to fibroblasts, and the expression of dermal markers altered by epidermal IL-1α production was evaluated.

[0173] Materials and methods

[0174] The reconstructed epidermis was topically treated with a formulation (cream) containing 1% tea tree oil concentrate or a placebo, or systemically treated with 1 μM dexamethasone (anti-inflammatory reference). After a 24-hour preincubation, the epidermis was stimulated with 0.5 μg / mL PMA and incubated again for 24 hours.

[0175] The culture supernatants were collected and subjected to IL-1α assays. On the other hand, these supernatants (conditioned medium) were placed on cultures of normal human dermal fibroblasts.

[0176] After incubation of fibroblasts in the presence of conditioned medium for 24 h, the expression of markers of interest was assessed by real-time quantitative PCR.

[0177] The results were statistically analyzed using ANOVA followed by Tukey's test.

[0178] result

[0179] Tea oil concentrate formulated at 1% significantly inhibited the release of IL-1α from PMA-induced reconstructed epidermis, an effect that was significantly different from that of placebo.

[0180] Administration of conditioned medium from PMA-stimulated reconstructed epidermis to fibroblast cultures induced changes in gene expression of inflammatory markers, oxidative stress, and synthesis and degradation of the dermal matrix. These results validate the "inflammatory aging" model.

[0181] Under these conditions, tea oil concentrate modulated the overexpression of cytokines and chemokines (CXCL1, IL8, IL6, CCL2) and matrix-degrading enzymes (MMP1, MMP3), and increased the gene expression level of HAS2.

[0182] These results suggest that tea oil concentrate can modulate the process of "inflammatory aging".

[0183] Table 12: Amount of IL-1α produced by reconstructed epidermis stimulated with PMA

[0184]

[0185] Table 13: Genes regulated by 1% tea oil concentrate formulation in "inflammatory senescence": Gene expression in fibroblasts incubated in the presence of conditioned medium

[0186] (Relative quantitative expression level; % protection compared to PMA)

[0187]

[0188] Active screening in skin reconstruction

[0189] Materials and methods

[0190] A formulation containing 1% tea tree oil concentrate (cream) or a placebo was applied to the surface of reconstructed human skin. After 24 hours of incubation, gene expression of markers of interest for dermal benefit was assessed by quantitative real-time PCR.

[0191] The results were statistically analyzed by Student's t test.

[0192] result

[0193] The 1% tea oil concentrate significantly increased the gene expression levels of the markers MKI67 and PSMD1. These effects were significantly different from those of placebo.

[0194] The two genes are involved in two different cellular functions: proliferation, marked by MKI67, and the proteasome, marked by PSMD1.

[0195] Aging is associated with a decrease in cell proliferation. Decreased expression of the marker MKI67 has been observed in aged cells. By increasing the expression of the MKI67 gene, tea oil concentrate can promote cell proliferation, demonstrating its anti-aging potential.

[0196] PSMD1 is a proteasome subunit that plays a key structural role by serving as a docking site for ubiquitinated substrates destined for clearance. Decreased levels of this marker have been observed in aged fibroblasts. A decreased ability of cells to clear damaged proteins is a hallmark of aging. By increasing PSMD1 gene expression, tea oil concentrate promotes the clearance of damaged proteins, demonstrating anti-aging potential.

[0197] Table 14: Genes significantly regulated by 1% tea oil concentrate formulation compared to placebo (set at 1.00)

[0198]

[0199] Barrier-promoting potential

[0200] Materials and methods

[0201] A formulation containing 1% tea tree oil concentrate (cream) or a placebo was applied to the surface of the reconstructed epidermis. After 24 hours of incubation, gene expression of markers of interest for epidermal benefit was assessed by quantitative real-time PCR.

[0202] The results were statistically analyzed by Student's t test.

[0203] result

[0204] Tea oil concentrate formulated at 1% significantly increased the gene expression levels of barrier markers (TJP1, DSC1, LOR, ZNF750, CERS3). These effects were significantly different from placebo, demonstrating the barrier-promoting potential of tea oil concentrate.

[0205] Table 15: Genes significantly regulated by 1% tea oil concentrate formulation compared to placebo (set at 1.00)

[0206]

[0207] Different types of cell junctions exist within the epidermis that ensure cohesion between keratinocytes: tight junctions, desmosomes, and gap junctions. These different junctions contribute to the skin's barrier function.

[0208] Tight junctions are intercellular connections that seal keratinocytes together. TJP1 protein is a marker of tight junctions. It is involved in controlling the paracellular diffusion of ions and solutes.

[0209] Desmosomes are localized connections between keratinocytes in the stratum basale, stratum spinosum, and stratum granulosum. Desmosomes consist of three parts: an intracellular portion, a transmembrane portion, and an extracellular portion. Desmocollin-1 is a component of desmosomes. It is a transmembrane glycoprotein belonging to the cadherin family.

[0210] Loricrin, ZNF50, and CERS3 markers are involved in the formation of the cornified envelope. During cornification, the plasma membrane of differentiating keratinocytes is replaced by the cornified envelope, which consists of insoluble, structured extracellular proteins covalently bound to each other and to remodeled lipids. This prevents excessive loss of body fluids and provides an effective barrier against pathogens.

[0211] About 80% of the cornified envelope is composed of loricrin.

[0212] Keratinocyte differentiation can be stimulated by various transcription factors such as ZNF50, a transcription factor known to regulate several genes encoding components of the cornified envelope as well as terminal differentiation of keratinocytes.

[0213] Ceramides are the main lipids in the skin and constitute approximately 50% of the intercellular lipids by mass. They are involved in the barrier function of the skin and the maintenance of hydration. Ceramide synthases (CERS) 3 and 4 are prominent in the skin.

[0214] Tea oil concentrate contributes to the barrier-promoting effect by significantly increasing the gene expression levels of these different markers.

[0215] Activity in in vitro models relevant to skin comfort and health

[0216] Psychological stress has multiple physiological and clinical effects on the skin. The physiological effects are diverse (inflammation, impaired barrier function, impaired healing, immunosuppression) and can lead to clinical effects (dry / sensitive skin phenotype, redness, irritation, itching, blemishes, acne, eczema, etc.).

[0217] There are two main axes of response to stress: the sympathetic nervous system, which induces the release of neuropeptides (such as substance P) and catecholamines, and the hypothalamic-pituitary-adrenal axis (HPA axis), which induces the release of cortisol.

[0218] Acute psychological stress can alter the permeability of the skin barrier and the integrity of the stratum corneum.

[0219] To evaluate the effects of tea oil concentrate on the biocomfort and well-being axis, different in vitro stress models involving cortisol and substance P were implemented.

[0220] Effects of tea oil concentrate on cortisol release in a skin explant model

[0221] Materials and methods

[0222] Skin explants were topically treated with a preparation (cream) containing 1% tea oil concentrate or a placebo thereof, or were systemically treated (in culture medium) with 0.001% tea oil concentrate and 1 mM of the positive reference metyrapone. The explants were incubated for 24 hours and then stressed with 0.6 μM ACTH (adrenocorticotropic hormone, a hormone that induces cortisol production). This stress was combined with explant cultivation in 42°C incubators to enable enhanced cortisol production. Explants were then systemically treated with a topical product or active ingredient and a positive reference. After incubation for 24 hours, the culture medium was recovered and an ELISA assay for cortisol was performed.

[0223] The results were statistically analyzed by Student's t test.

[0224] result

[0225] Tea oil concentrate tested systemically in culture media reduced Cortisol in the supernatant of explant cultures (Cortisol induced by ACTH treatment combined with explant culture in a 42°C incubator).

[0226] A 1% tea tree oil concentrate suppressed cortisol compared to a placebo, which increased cortisol release.

[0227] Table 16: Amount of Cortisol Produced by Skin Explants Stimulated with ACTH + T°C 42°C (Tea Oil Concentrate Tested Systemically in Culture Medium)

[0228]

[0229] Table 17: Amount of Cortisol Produced by Skin Explants Stimulated with ACTH + T°C 42°C (Tea Oil Concentrate Topically Tested in Formulations Compared to Placebo)

[0230]

[0231] Effects of tea oil concentrate in an exogenous / endogenous stress model related to skin barrier protection

[0232] Tea oil concentrate was evaluated in a keratinocyte model (HaCaT) that was stressed with cortisone and then UVA. UVA stress enhances the conversion of cortisone (an inactive molecule) to cortisol (an active molecule). Cortisol is known to alter the skin barrier.

[0233] Materials and methods

[0234] HaCaT keratinocytes were treated with 0.0005% and 0.001% tea oil concentrate and 100 nM cortisone. After 24 hours of incubation, the cells were exposed to 3 J / cm 2 Cells were stressed by irradiation with 100 μg of UVA. Cell lysates were collected immediately after irradiation for analysis of carbonylated proteins by immunostaining or 6 h after irradiation for analysis of barrier and inflammatory markers by RT-qPCR.

[0235] The results were statistically analyzed using ANOVA followed by Dunnett's test.

[0236] result

[0237] Cortisol stress combined with UVA increased IL-1α gene expression, reflecting inflammatory activation. It also increased the number of carbonylated proteins. It suppressed gene expression of barrier markers (loricrin, filaggrin, and keratin 10), demonstrating the negative impact of cortisol on the skin barrier.

[0238] Tea oil concentrate showed a pro-barrier protective effect by increasing the markers of cortisol / UVA stress suppression: filaggrin, loricrin, and keratin 10. Tea oil concentrate also showed an anti-inflammatory effect on IL1a (gene expression) and reduced the number of stress-induced protein carbonyls (damaged proteins).

[0239] This latter result can be correlated with those observed during the active screening on reconstructed skin, where tea oil concentrate significantly increased the gene expression levels of the marker PSMD1 (26S proteasome non-ATPase regulatory subunit 1, a proteasome marker involved in the degradation of damaged proteins).

[0240] Table 18: Genes significantly regulated by tea oil concentrate (QR = relative quantification - untreated control set as 100)

[0241]

[0242] Table 19: Quantification of carbonylated proteins (QR = relative quantification)

[0243]

[0244] Effects of tea oil concentrate on the cortisol stress model affecting the skin barrier

[0245] The studies were conducted on a skin explant model in which the barrier was weakened by tape stripping and subsequently stressed by cortisol, which damages the skin barrier and delays its repair.

[0246] Materials and methods

[0247] The stratum corneum of skin explants was altered by tape stripping and subsequent 0.001% cortisol stress. After 6 hours of incubation, the surface of the explants was treated with a formulation (cream) containing 1% tea tree oil concentrate or a placebo thereof. 24 hours after the first application (D1), the explants were treated again with cortisol and then 6 hours later with the product. These treatments (cortisol + product after 6 hours) were repeated for a total of 5 treatments (D0 to D4). 24 hours after the last treatment (D5), the explants were recovered for different markings:

[0248] - Hematoxylin / eosin labeling for stratum corneum thickness analysis,

[0249] - Safranin-O-red marker for analyzing the number of stratum corneum layers,

[0250] -Lucifer Yellow labeling for skin barrier integrity analysis.

[0251] The results were statistically analyzed using ANOVA followed by Tukey or Dunnett's test.

[0252] result

[0253] Alterations in the stratum corneum by tape stripping followed by repeated treatment with cortisol over several days significantly reduced stratum corneum thickness and skin barrier integrity.

[0254] Compared to placebo, the tea oil concentrate tested at 1% showed a significant effect on the thickness of the stratum corneum and the number of stratum corneum layers (histological markers), with the stratum corneum being thicker and having more layers in the case of treatment with the active ingredient. It also showed a significant effect on the integrity of the skin barrier (penetration of the fluorescein marker).

[0255] Therefore, tea oil concentrate promotes the repair of the skin barrier and protects against psychological stress that alters skin barrier repair.

[0256] Table 20: Quantification of stratum corneum thickness

[0257]

[0258] Table 21: Quantification of the number of stratum corneum layers

[0259]

[0260]

[0261] Table 22: Analysis of skin barrier integrity by permeability of fluorescent Lucifer yellow marker

[0262]

[0263] These diverse results suggest that tea oil concentrate is highly beneficial for the comfort of dry and sensitive skin that may be caused and aggravated by psychological stress.

[0264] Effects of tea oil concentrate in a model of epidermal re-innervation

[0265] Materials and methods

[0266] Human iPS (induced pluripotent stem) cells are differentiated into sensory neurons in culture inserts. Collagen gel is then applied to the sensory neurons. After 3 weeks of culture, the explanted epidermis separated from the dermis by dispase treatment is placed on the collagen gel. The model is incubated for 5 days (the time it takes for the epidermis to be re-innervated by sensory neurons). The epidermis is then topically treated with 0.5% and 1% tea oil concentrate formulated in paraffin. After incubation for 24 hours, the epidermis is topically treated with 10% lactic acid for 15 minutes. The culture medium is then recovered and an ELISA assay for substance P is performed.

[0267] The results were statistically analyzed using the Mann-Whitney U test.

[0268] b. result

[0269] Tea oil concentrate tested at 0.5% and 1% significantly inhibited the release of substance P, a neuropeptide involved in pain perception, induced by lactate in a re-innervated epidermis model.

[0270] Therefore, tea oil concentrate is beneficial for the comfort of sensitive skin.

[0271] Table 23: Amounts of Substance P Released by Reinnervated Epidermis Stressed with Lactic Acid

[0272]

[0273]

[0274] By reducing the release of cortisol and substance P and by protecting the skin from the harmful effects of cortisol, tea oil concentrate brings comfort and well-being to dry and sensitive skin.

[0275] Example 3: In vivo bioactivity test of the extract according to the present invention

[0276] Concept and plan

[0277] Conception

[0278] People with dry and / or sensitive skin experience skin symptoms (stinging, tightness, etc.), which negatively impact their skin comfort and well-being, potentially leading to psychological stress or worsening emotional states and also to a decrease in their quality of life. A vicious cycle can then begin, with psychological stress itself becoming the root cause of skin problems (Farage, 2022).

[0279] In the context of this problem, the benefit of cosmetic or dermatological active ingredients is to act on the symptoms associated with dry and / or sensitive skin, to limit the negative impact on skin discomfort and on the quality of life regarding people's psychological state, and to improve their emotional state and well-being.

[0280] plan

[0281] Purpose

[0282] The aim of this study was to evaluate the efficacy and emotional benefits of the extract according to the invention compared to placebo in people experiencing psychological discomfort / stress / bad mood due to dry and / or sensitive skin.

[0283] product

[0284] The extract tested was the concentrate from Example 1. The active ingredient was formulated with 1% tea oil concentrate. Details of the composition are shown in the table below.

[0285] Table 24:

[0286]

[0287] c. parameter

[0288] - Application for 28 days, twice a day (morning and evening), face and body (assessment on the face).

[0289] - Active comparison study (n=22, 18 dry skin / 19 sensitive skin, 51.3±14.7 years [19 to 70]) versus placebo (n=22, 19 dry skin / 20 sensitive skin, 45.3±13.8 years [23 to 68]), double-blind, randomized.

[0290] - Healthy (no skin or other unstable pathologies that could interfere with the study, no symptoms during the exploratory assessment period) female subjects, older than 18 years - Phototype I to IV (Caucasian).

[0291] - Subjects with dry and / or sensitive skin type on the face (associated with functional and / or clinical signs) (evenly distributed between dry and sensitive skin).

[0292] - Subjects who experience psychological discomfort / stress / bad mood due to dry skin and / or sensitive skin.

[0293] -Evaluated using a combination of clinical assessment, subjective questionnaire evaluation, and objective physiological measurements.

[0294] result

[0295] Skin sensitivity ("Sensitivity Scale")

[0296] The "Sensitivity Scale" described in the literature assesses the degree of overall skin irritation and the severity of skin conditions (Misery et al., 2014). This scale consists of ten items corresponding to the sensations (eight items) or appearances (two items) of signs associated with skin discomfort: stinging, burning, warmth, tightness, itching, pain, general discomfort, hot flashes, redness, and peeling.

[0297] Subjects can rate the intensity of each parameter from 0 to 10.

[0298] For each of the ten parameters, the improvement between T0 and T28 was significant with the active ingredient, whereas only six parameters were significantly improved with placebo (see Figure 1 ).

[0299] The active ingredient showed significantly improved results compared to placebo. In fact, the following three parameters were at the threshold of p < 0.05: stinging, burning and itching. In addition, the following three parameters and the total score were at the threshold of p < 0.1 (see Figure 2 ): A feeling of warmth, tightness, and general discomfort.

[0300] Rhythm and Heart Coherence

[0301] Prosody corresponds to the emotional content of the sound and can be assessed by studying the sound spectrum, revealing variations in several parameters. Algorithmic processing is used to quantify the sound stress load.

[0302] After 28 days, the active ingredient significantly reduced sound stress, with the change being significantly better than placebo (see Figure 3 ).

[0303] Heart rate variability (HRV) is a cardiac measure of the interaction between the sympathetic and parasympathetic branches of the autonomic nervous system (ANS). Slight variations in HRV are often used in psychosocial studies of stress because they provide one of the most stable parameters of a subject's emotional strain. Stress and negative emotions are known to disrupt cardiac coherence and lead to irregularities in the HRV signal. Therefore, more positive emotions should increase HRV index values.

[0304] After 28 days, the active ingredient significantly improved HRV cardiac coherence, with the difference being significantly better than placebo (see Figure 4 ).

[0305] Galvanic skin response

[0306] The galvanic skin response (EDR) corresponds to a change in skin resistance when an electric current is passed through it. It is a reaction that occurs after sensory or emotional arousal. This change in skin resistance is the result of physiological changes that may be unconscious or imperceptible to the naked eye. These autonomic nervous system changes are the result of activation of the sympathetic nervous system. Indeed, after psychological (i.e., emotional) arousal, sweat gland activity increases, which in turn increases skin conductance, causing a very rapid EDR. This reaction occurs primarily in the plantar and palmar regions. Sweat glands are controlled by the sympathetic nervous system via fibers from cholinergic ganglia. In this way, the galvanic skin response can be used as a physiological sign of emotional arousal.

[0307] In this study, after attaching electrodes for galvanic skin response (EDR) to their fingers, technicians asked the subjects to cover their eyes with an eye mask. Before heating, a patch inducing temperature stimulation was placed on the subject's cheek. After a 2-minute rest period to stabilize parameters, technicians triggered the patch to heat for 30 seconds to achieve temperature stimulation and record the galvanic skin response (EDR). The cheek patch was then removed (the subject did not see the patch at any time).

[0308] After 28 days, both the active ingredient and the placebo significantly reduced the galvanic skin response after thermal stimulation, with the changes by the active ingredient being significantly better compared to the placebo (see Figure 5 ).

[0309] Example 4: In vivo bioactivity test of the extract according to the present invention

[0310] Concept and plan

[0311] 2) Conception

[0312] The purpose of this study is to evaluate the efficacy and benefits of an active ingredient compared to placebo on skin conditions in people with dry and / or sensitive skin conditions.

[0313] The skin barrier plays a regulatory role in the skin's water balance. When the skin barrier is damaged, the regulation of water exchange is disrupted. Water then migrates more easily to the external environment, increasing transepidermal water loss (TEWL). On the other hand, if the skin barrier condition improves, water loss values ​​decrease because proper regulation of water exchange is ensured.

[0314] plan

[0315] Purpose

[0316] The purpose of this study is to evaluate the efficacy and benefits of an active ingredient compared to placebo on skin conditions in people with dry and / or sensitive skin conditions.

[0317] product

[0318] The extract tested was the concentrate from Example 1. The active ingredient was formulated from 1% tea oil concentrate. Details of the composition are shown in the table below.

[0319] Table 25:

[0320]

[0321]

[0322] parameter

[0323] - Application for 28 days, twice a day (morning and evening), face and body (assessment on the face).

[0324] - Comparative study of active ingredient (n=24, 19 obviously dry skin / 23 sensitive skin, 50.0±16.7 years [24 to 71]) versus placebo (n=24, 22 obviously dry skin / 22 sensitive skin, 54.2±16.8 years [24 to 74]), double-blind, randomized.

[0325] - Healthy (no skin or other unstable pathologies that could interfere with the study, no symptoms during the exploratory assessment period) female subjects, older than 18 years - Phototype I to IV (Caucasian).

[0326] - At least 50% of the subjects had noticeable dry skin on the face and at least 50% of the subjects had sensitive skin (assessed according to the questionnaire in the literature: Boyer et al., 2021), with one clinical sign and one subjective symptom under at least two stressors.

[0327] - The subject experienced stinging and / or warming sensations, burning and / or itching.

[0328] - Subjects exhibiting skin complaints ≥ 4 / 10.

[0329] -Evaluation: Using Tewameter™ Transepidermal water loss is measured. A probe consisting of two sensors is passed through a stream of water vapor. The partial pressure difference between the two sensors is measured. This value corresponds to the evaporation rate of the volatile substance (in this case, water).

[0330] Hydration and barrier function results

[0331] Hydration was significantly improved by both the active ingredient and the placebo.

[0332] The skin barrier, as assessed by TEWL, was significantly improved by the active ingredient and placebo (see Figure 6 The difference between the active ingredient and the placebo was significant in favor of the active ingredient (Δ% (Δ active ingredient / Δ placebo) = -34.8%, p < 0.05 (Mann-Whitney test)).

[0333] References:

[0334] Yang C., Liu

[0335] Hu JB,Yang GLPhysiochemical characteristics,fatty acid profileand tocopherol composition of the oil from Camellia oleifera Abel cultivatedin Henan,China,Grasas y aceites 2018,69(2)

[0336] Wen Y.,Su S.-C.,Lv-yi M.,Yang S.-Y.,Wang Y.-W.,Wang X.-N., Effects of canopy microclimate on fruit yield and quality of Camelliaoleifera.ScientiaHorticulturae,2018,235.https: / / doi.org / 10.1016 / j.scienta.2017.12.042.

[0337] Farage MAPsychological Aspects of Sensitive Skin:AViciousCycle.Cosmetics2022,9,78.https: / / doi.org / 10.3390 / cosmetics9040078

[0338] Misery L.,Jean-Decoster C.,Mery S.,Georgescu V.,Sibaud V.Anew ten-itemquestionnaire for assessing sensitive skin:the Sensitive Scale-10.ActaDermVenereol.2014,94(6):635-9.doi:10.2340 / 00015555-1870.

[0339] Boyer G.,Belilovsky C.D.,BrédifS.,Baudouin C.,Misery L.,BellemèreG.Clinical and Instrumental Exploration of Sensitive Skin in aPediatricPopulation.Cosmetics 2021,8,43.https: / / doi.org / 10.3390 / cosmetics8020043。

Claims

1. A lipid extract of tea oil, characterized in that The lipid extract is Camellia oleifera tea oil concentrated into an unsaponifiable fraction, which contains 1.5 to 100 weight percent, preferably 1.8 to 80 weight percent, and preferably 2 to 60 weight percent of unsaponifiable matter relative to the total weight of the extract.

2. The extract according to claim 1, characterized in that The lipid extract has a sterol content greater than 1.5% by weight, preferably 2% by weight, relative to the total weight of the extract.

3. The extract according to claim 1 or 2, characterized in that The lipid extract preferably comprises 10% to 50% by weight, preferably 15% to 40% by weight, more preferably 15% to 35% by weight of β-amyrin and Δ5-avenasterol, relative to the total weight of sterols, and / or preferably 10% to 50% by weight, preferably 15% to 40% by weight, more preferably 15% to 35% by weight of lupeol and Δ7-stigmasterol, relative to the total weight of sterols.

4. A method for preparing the extract according to any one of claims 1 to 3, comprising the following consecutive steps: a) molecular distilling the crude oil or refined oil of Camellia oleifera; b) extraction of unsaponifiable matter, where applicable; c) recovering the concentrated unsaponifiable oil obtained after step a) or the unsaponifiable matter obtained after step b); and d) optionally a step of deodorizing and / or decolorizing the concentrated unsaponifiable oil or the unsaponifiable matter.

5. The method according to claim 4, characterized in that Step b) comprises the following consecutive steps: i. after step a) the tea oil concentrated into the unsaponifiable fraction obtained is saponified, ii. The unsaponifiable matter is then extracted using a suitable solvent such as ethyl acetate and hexane.

6. A composition comprising as an active ingredient the lipid extract of Camellia oleifera oil according to any one of claims 1 to 3, and a suitable excipient.

7. The composition according to claim 6, characterized in that It comprises from 0.01% to 20% by weight, advantageously from 0.05% to 15% by weight and more advantageously from 0.1% to 10% by weight of said lipid extract relative to the total weight of the composition.

8. A composition according to claim 6 or 7, or an extract obtainable by the process according to claim 4 or 5, or an extract according to any one of claims 1 to 3, for use in a dermatological composition, a pharmaceutical composition, a nutraceutical composition or as a food supplement.

9. Composition or extract according to claim 8, for use in the prevention and / or treatment of diseases or conditions of the skin and / or mucous membranes and / or appendages, or as a healing agent, said diseases or conditions being advantageously inflammatory reactions, diseases associated with endogenous or exogenous stress, such as psychological stress, stress associated with anxiety states, stress associated with free radical attacks with or without chemical or atmospheric pollution and / or exposure to UV or IR, disorders of the barrier or homeostasis, photosensitive skin, mechanical and / or thermal aggression.

10. The composition or extract according to claim 9, for use in preventing and / or treating dry skin, xerosis, sensitive skin and irritation.

11. Composition or extract according to claim 9, for use in the prevention and / or treatment of conditions or diseases selected from the group consisting of epidermal scars, delicate lips and cheilitis, stretch marks, skin after bites, skin abrasions, spots and / or skin scabs, and delicate and sensitive skin, advantageously selected from the group consisting of delicate and sensitive skin.

12. Cosmetic use of an extract according to any one of claims 1 to 3 or an extract obtainable by a process according to claim 4 or 5 or a composition according to any one of claims 6 or 7, in a form suitable for topical or oral administration, for improving the condition and / or appearance of healthy skin and / or healthy appendages and / or healthy mucous membranes, advantageously for improving the firmness, elasticity or firmness of healthy skin and / or for improving skin comfort.

Images