Cosmetic uses of a hydrolyzed extract of Silybum marianum seed cake

Abstract

Cosmetic Uses of a Hydrolyzed Extract of Silybum marianum Seed Cake: The invention relates to the cosmetic use of a hydrolyzed extract of Silybum marianum seed cake to maintain and / or enhance the biomechanical properties of skin appendages. Another object of the invention is a cosmetic treatment method comprising the topical application of the hydrolyzed seed cake extract or a composition containing it.

Description

Title of the invention: Cosmetic uses of a hydrolyzed extract of Silybum marianum seed cake. Technical field

[0001] The invention relates to cosmetic uses of a hydrolyzed extract of Silybum marianum seed cake. Previous technique

[0002] The care of skin appendages such as nails, hair, eyelashes, eyebrows, involves a global treatment, that is to say a direct treatment of the keratin fiber but also of the corresponding skin area, including the scalp and hair follicles (or hair bulbs and hair fibers).

[0003] Furthermore, direct hair fiber care involves treating the proteins that constitute it. Hair proteins, such as keratins, are particularly sensitive to damage induced by lifestyle, exposure to hygiene chemicals (including their type and frequency of use), styling habits, and environmental conditions (sun, pollution, salt, wind, climatic variations), general lifestyle habits, and age. These proteins become denatured, making the hair follicles less resistant, less supple, more brittle, and duller, resulting in split ends. These follicles then lose their biomechanical properties.

[0004] One solution is the application of exogenous hydrolyzed proteins to compensate for the damage to keratin proteins and to improve the hydration and elasticity of keratin fibers, particularly hair fibers. These proteins thus give the fibers more softness, shine, bounce, and body.

[0005] The hydrolysis (chemical, thermal, or enzymatic) of these proteins is a necessary preliminary process to fragment them into low molecular weights, thereby promoting their water solubility and penetration into the keratin fiber, and facilitating their formulation. Only molecules with a low average molecular weight (less than 7 kDa) can penetrate the keratin fiber, particularly damaged hair fibers. They strengthen the keratin fiber from within, making it stronger, more elastic, less brittle, and, in particular, reducing split ends in the hair fiber.

[0006] High-performance hydrolyzed proteins exhibit adhesion properties to the hair fiber (weak bonds with hair keratins) due to ionic charges and polar sites (interactions via van der Waals forces). They can also form protective films on the surface of the hair fiber.

[0007] Products based on hydrolyzed proteins already exist on the cosmetics market. These proteins are of plant or animal origin. The most common are hydrolyzed proteins of wheat, silk, keratin, collagen, elastin, milk, or almonds. However, there is a constant need for high-performance alternative ingredients in this field.

[0008] Quite unexpectedly, the Applicant discovered that a hydrolyzed extract of Silybum marianum seed cake had the ability to penetrate the keratin fiber, thereby enhancing its biomechanical properties. This hydrolyzed extract proved capable of improving the quality of skin appendages, increasing their resistance and reducing breakage, particularly that of hair, making it shinier, more radiant, and fuller. Furthermore, the hydrolyzed extract of Silybum marianum seed cake possesses skin appendage repair properties, as will be demonstrated later in this description.

[0009] One advantage of the hydrolyzed extract according to the present invention is that it is a co-product derived from an industrial extraction residue that is usually not recovered and is considered industrial waste, thus making this invention part of an eco-responsible approach. Another advantage is that it is an ingredient that provides comprehensive care, as it has effects on both the keratin fiber, particularly the hair, and the hair follicles. Furthermore, the hydrolyzed extract according to the invention can be easily produced on an industrial scale. Finally, enzymatic hydrolysis makes it possible to fractionate the proteins in the press cake to recover low molecular weight peptides, precisely those capable of penetrating the keratin fibers to ensure the cosmetic effects described herein.

[0010] The hydrolyzed extract according to the invention is derived from the press cake of the plant Silybum marianum. This plant, also known as milk thistle, has been known for millennia for its liver-supporting properties. It is still used in Asia and Africa to treat kidney problems, gastrointestinal disorders, heart disease, rheumatism, and fever. It is primarily the seeds that are used for these therapeutic purposes.

[0011] The young stems, seeds, and fleshy shoots are still traditionally consumed in several countries of the Middle East, North Africa, Sardinia, and Spain. The fruits of Silybum marianum can be used either for oil extraction or as flour, mixed with wheat flour, for the preparation of various baked goods or as food products.

[0012] Silybum marianum is also used generally in cosmetics, in particular the use of the flowers (without the seeds) to improve the elasticity, firmness, barrier effect and hydration of the skin, as well as to reduce hair loss.

[0013] Application DE 4323614 discloses a Milk Thistle fruit extract used as a hair growth and regeneration agent.

[0014] Application CN103446021 discloses an extract of Silybum marianum seeds obtained by heating and propylene glycol extraction of whole seeds protecting hair from UV damage with an antioxidant action that protects hair color but also improves firmness and tensile strength.

[0015] French patent FR3053253 describes a process for preparing an extract of Silybum marianum achenes comprising less than 0.2% silymarin. The process comprises oil extraction with a solvent including hydrotropic aqueous solution, subcritical water, ethanol, isopropanol, or mixtures thereof.

[0016] Thus, to the applicant's knowledge, no prior art discloses the cosmetic uses of the hydrolyzed extract according to the present invention. Nor does any document, alone or in combination, suggest them. Description of the invention

[0017] A first object thus relates to the non-therapeutic cosmetic use of a hydrolyzed extract of Silybum marianum seed cake to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

[0018] A second object relates to the non-therapeutic cosmetic use of the hydrolyzed extract according to the invention in a cosmetic composition.

[0019] A third object relates to a non-therapeutic cosmetic care process comprising the topical application of the hydrolyzed extract according to the invention or of a cosmetic composition comprising it to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

[0020] A first object therefore relates to the non-therapeutic cosmetic use of a hydrolyzed extract of Silybum marianum seed cake to maintain and / or increase the biomechanical properties of skin appendages advantageously of keratin fibers, preferably of hair.

[0021] The term "hydrolyzed extract" means the product obtained after enzymatic or chemical hydrolysis in an aqueous solvent of the seed cake of Silybum marianum. In a preferred embodiment, the hydrolyzed extract is obtained by enzymatic hydrolysis.

[0022] The term "oilcake" refers to the co-product of seed extraction, that is, the residue obtained after oil extraction. Preferably, the oilcake of seeds Silybum marianum used is a delipidated cake of Silybum marianum seeds.

[0023] The term “delipidated” means an oil content of the cake strictly less than 10% by weight, preferably strictly less than 8% by weight, relative to the total weight of the cake.

[0024] Generally speaking, "oil" means all lipophilic compounds such as fat-soluble vitamins, fatty acids, mono-, di- and triglycerides and phospholipids contained in the seed of Silybum marianum

[0025] The term "seeds" refers to the fruit or achene without its covering. This covering, also called the pericarp, is therefore not contained within the seeds according to the invention.

[0026] “Cosmetic use” means a non-therapeutic, non-pharmaceutical and non-dermatological use, that is to say, one which does not require and does not involve therapeutic treatment and is intended for healthy skin appendages and / or healthy skin with skin appendages.

[0027] The term “healthy” means skin appendages and / or skin that are described as non-pathological by a specialist in the field, a dermatologist, that is to say, that do not present candidiasis, diseases or conditions, infection, inflammation, scar, skin disease or condition such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, in particular seborrheic dermatitis, dandruff, or sores or injuries and / or other dermatoses and / or alopecia and / or baldness and / or alopecia areata.

[0028] Thus, for the purposes of the invention, "damaged" skin appendages are those classified as "non-pathological" by a specialist in the field. They are therefore healthy despite being damaged. In particular, damaged skin appendages are those that have lost their suppleness and / or plasticity and / or resistance and / or strength and / or shine and / or color and / or volume and are, consequently, particularly dehydrated. "Damaged" skin appendages have thus lost their biomechanical properties. As a result, they become dull, dry, less resistant, friable, fragile, brittle, split, curl, and / or are forked.Damaged skin appendages can result from exposure to stressors such as environmental conditions like wind, cold, salt, chlorine, pollution, sun, lifestyle or age, but also from exposure to chemical or mechanical aggressions due to daily treatments of the appendages such as friction, styling, curling, coloring, bleaching, straightening, hair drying, cosmetic or hygiene chemicals such as nail polish, makeup and / or makeup remover, shampoos, conditioners or creams.

[0029] Keratin fibers, particularly hair, are also more difficult to Styling and shaping are difficult. This loss of surface quality in the hair is visible and unsightly. The hair reflects less light and is therefore noticeably less shiny, less colored, and less luminous. It is also thinner and less thick. This is particularly true for colored or highlighted hair.

[0030] The term "skin appendages" herein refers to hair, eyelashes, eyebrows, body hair, including beard hair, and / or nails. According to the invention, the beard includes the mustache. Preferably, it refers to hair.

[0031] The term "keratin fibers" refers to all the fibers constituting the human hair system, including head hair, eyelashes, eyebrows, and body hair, particularly beard hair, including the moustache. Preferably, this refers to head hair.

[0032] The hydrolyzed extract according to the invention is a topically acceptable ingredient.

[0033] The term "topically acceptable" means an ingredient suitable for topical application, non-toxic, non-irritating to the skin, in particular the scalp or skin appendages, not inducing an allergic or inflammatory response, and not chemically unstable.

[0034] The hydrolyzed extract according to the invention can be used topically.

[0035] The term “topical route” means the direct local application and / or spraying of the ingredient onto the surface of the skin, in particular the scalp, or skin appendages, in particular the hair.

[0036] The hydrolyzed extract can be applied topically to all or part of the skin surface including skin appendages, in particular the scalp, or to all or part of the skin appendages, advantageously on nails, hair, body hair, in particular beard hair, eyelashes and / or eyebrows, more advantageously on keratin fibers, even more advantageously on hair.

[0037] An object of the invention thus relates to the non-therapeutic cosmetic use of a hydrolyzed extract of Silybum marianum seed cake to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, more advantageously hair and / or to maintain and / or increase their biomechanical properties.

[0038] For the purposes of the present invention, "repairing biomechanical properties" means improving and / or at least partially increasing the biomechanical properties of skin appendages damaged by exposure to environmental, chemical, biological, and / or mechanical stresses, preferably keratin fibers, advantageously hair. In particular, repaired skin appendages can regain the biomechanical properties they had before being damaged, that is, those of undamaged skin appendages. It is therefore possible to compare the effect of the hydrolyzed extract according to the invention on these biomechanical properties by comparing, for example, the biomechanical properties of skin appendages before and after treatment with the hydrolyzed extract according to the invention or by comparing the biomechanical properties of damaged skin appendages after treatment with the hydrolyzed extract according to the invention to the biomechanical properties of undamaged skin appendages.

[0039] For the purposes of this invention, "maintaining biomechanical properties" means preventing and / or avoiding the degradation of the biomechanical properties of skin appendages, preferably keratin fibers, advantageously hair, particularly when exposed to environmental, chemical, biological and / or mechanical aggressions, notably by preventing skin appendages from becoming damaged skin appendages and / or by preventing these skin appendages from becoming even more damaged.It is therefore possible to compare the effect of the hydrolyzed extract according to the invention on these biomechanical properties by comparing, for example, the biomechanical properties of skin appendages treated with the hydrolyzed extract according to the invention and exposed to environmental, chemical, biological and / or mechanical aggressions to the biomechanical properties of skin appendages not treated with the hydrolyzed extract according to the invention and exposed to environmental, chemical, biological and / or mechanical aggressions.

[0040] For the purposes of this invention, "increasing biomechanical properties" means improving the biomechanical properties of skin appendages, preferably keratin fibers, advantageously hair, particularly skin appendages damaged by exposure to environmental, chemical, biological, and / or mechanical stresses, but also undamaged skin appendages. It is therefore possible to compare the effect of the hydrolyzed extract according to the invention on these biomechanical properties by comparing, for example, the biomechanical properties of the skin appendages before and after treatment with the hydrolyzed extract according to the invention.

[0041] For the purposes of this invention, "biomechanical properties" means the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e. their non-brittle appearance, and / or flexibility of skin appendages, preferably keratin fibers, advantageously hair.

[0042] Advantageously, the use of a hydrolyzed extract of Silybum marianum seed cake is to repair skin appendages, preferentially keratin fibers, advantageously hair.

[0043] Preferably, the use of a hydrolyzed extract of Silybum marianum seed cake is for maintaining and / or increasing the resistance and / or strength and / or volume and / or color and / or shine and / or plasticity, i.e. the non-brittle appearance, and / or flexibility of skin appendages, preferably keratin fibers, advantageously hair.

[0044] These properties can be evaluated ex-vivo by mechanical tests where is measured The resistance of skin appendages, preferentially keratin fibers, and more specifically hair, to tensile, stretching, bending, frictional, and torsional forces. These biomechanical parameters, particularly those of hair, can be evaluated in response to stretching and can be measured, for example, by the tensile test (Dia-Stron). The measured parameters can be the elastic modulus (Pa), elongation at break (%), breaking force (gmf), and the gradient of the plastic deformation region (gmf / % elongation), and are normalized with respect to the diameter of the skin appendages. This last parameter (gradient) allows the measurement of the plasticity of the material under study just before breakage. The technique makes it possible to obtain a stress (or force) curve as a function of the characteristic elongation of the material under study.It is known that an increase in elongation at break and a decrease in the plastic deformation gradient can be observed on capillary fibers (skin appendages) damaged by oxidation, reduction or ultraviolet irradiation following the breaking of disulfide bonds in free groups in the cortical domain.

[0045] In one embodiment of the invention, "maintaining and / or increasing the biomechanical properties" of skin appendages means reducing the advanced glycation end products (AGEs) that appear during chrono-induced or photo-induced aging, in particular, but also those that appear daily in response to environmental conditions, upon contact with oxidative chemical or physical agents. These glycation products reduce the biomechanical properties of skin appendages.

[0046] Environmental stressors include, in particular, smoke, especially from cigarettes, pollution, especially metals, PM2.5 and PM10 fine particles, temperature, especially heat and cold and their abrupt variations, water content, especially humidity or dryness, solar radiation, especially visible spectrum, UV and / or gamma rays and / or blue light, rain, wind, dust, and sea salt, swimming pool water, especially chlorine and transition metals present in the water, but also intrinsic and / or chrono-induced aging. The latter is indeed also responsible for the loss of the structural and / or functional qualities of skin appendages, preferentially keratin fibers, in particular eyelashes, eyebrows and hair, more particularly hair.

[0047] Chemical aggressions include chemical agents such as aggressive convenience, hygiene and beauty products, in particular aggressive shampoos and hair care and / or treatments, especially for styling, shaping such as straightening and / or perms and / or for coloring and / or bleaching, chlorine and transition metals present in swimming pool water, varnishes and / or solvents, solvents, makeup products.

[0048] Mechanical aggressions include, in particular, friction such as brushing and / or friction on fabrics (pillows, clothes) and / or particles such as dust and / or sand, heat from hair dryers and straighteners, and / or styling, including exposure to tensile, stretching and / or twisting forces.

[0049] Thus, in one embodiment of the invention, the ability of the extract according to the invention to "repair" damaged skin appendages can be evaluated using existing techniques in the field. Conventional methods for measuring a repair effect make it possible to measure the ability of the evaluated product to restore a visual, structural, and / or functional condition of a damaged skin appendage, preferably damaged hair, comparable to the condition of an undamaged skin appendage, preferably undamaged hair. These methods are performed on damaged (or damaged) skin appendages, preferably hair, and the repair effect is measured by comparison to undamaged (or damaged) skin appendages, preferably hair. Advantageously the repairing effect is a repairing effect of skin appendages damaged (or damaged) in vitro by an oxidizing agent, which induces a denaturation of the proteins of the appendages.Advantageously, this refers to a repairing effect of the hydrolyzed extract according to the invention on hair, and preferably the oxidizing agent is hydrogen peroxide. Further advantageously, this protein denaturation is evaluated by measuring the protein denaturation temperature (°C) by differential calorimetry (Wortmann and Deutz, Appl. Polym. Sci., 48, 137-150 (1993)) using a differential enthalpy analyzer (DSC Q100, TA Instruments), under the conditions described in Example 10.

[0050] To evaluate this repair effect of skin appendages in vivo, advantageously hair, several methods can be used, including video microscopy, confocal microscopy, FTIR or Raman microscopy, X-rays, and electron microscopy, which aims in particular to observe the condition and / or quality of the cuticle, the protective covering of the keratin fiber of skin appendages. Microscopy studies on whole hair allow the visualization and quantification of detached scales as an indicator of surface damage. These methods can also be used on hair sections to observe the condition and / or quality of the keratin fiber cortex.Physical surface quantification methods can be used to assess the morphology of skin appendages, particularly hair, such as Atomic Force Microscopy or White-light interferometric pro-filometry, and to evaluate chemistry (XPS), charge (Streaming potential) or energy (inverse Gas Chromatography).

[0051] To evaluate the repair effect of the hydrolyzed extract according to the invention on the internal properties of skin appendages, in particular hair, digital scan calorimetry (DSC) can be used.

[0052] For the purposes of the present invention, "maintain and / or increase volume" means maintaining and / or increasing the thickness of the skin appendages along their entire length, preferably maintaining and / or increasing the diameter of the keratin fibers, advantageously the diameter of the hair.

[0053] For the purposes of this invention, "maintaining and / or increasing the biomechanical properties" of skin appendages means maintaining and / or increasing their resistance, gloss, color, volume, strength, and / or plasticity, i.e., their non-brittle appearance and / or flexibility. These properties can be measured by the following methods and described in the following paragraphs and corresponding examples: - by measuring anti-radical activity, particularly in-tubo as described in example 3; - by measuring the chelating activity of transition metals such as iron (Fe), particularly in-tubo as described in example 4; - by measuring the chelating activity of transition metals such as copper (Cu), particularly in-tubo as described in example 5; - by measuring anti-glycation activity on proteins of skin appendages, particularly in vitro as described in example 6; - by measuring the carbonylation of keratins and associated proteins of skin appendages, particularly ex vivo, as described in examples 7a and b); - ex vivo by measuring the hair protection score (SPC) as described in example 7d; - ex vivo by measuring the stabilization of cysteine ​​groups as in example 9; - ex vivo by tensile strength tests (Diastron), fatigue tests, or repeated hair styling tests to assess breakage such as in examples 11 and 13; - ex vivo by measuring color variations in skin appendages such as in example 12; - in vitro by measuring the amount of ATP in the fibroblasts of the papilla of hair follicles as in example 14.

[0054] These properties can be evaluated in-tubo by measuring the anti-radical activity, for example by the DPPH test, measured in the presence of the hydrolyzed extract according to the invention. Thus, in a particularly preferred embodiment, the hydrolyzed extract according to the invention is effective in the quantity required to "maintain and / or increase the biomechanical properties" of skin appendages when the percentage of inhibition of DPPH° free radical oxidation in the presence of the hydrolyzed extract according to the invention, under the conditions described in Example 3, is greater than 50%. Advantageously greater than 65%, more advantageously greater than 80%, preferably greater than 90%, relative to the percentage of inhibition of DPPH° free radical oxidation, measured in the absence of the hydrolyzed extract according to the invention. Advantageously, the skin appendages are the keratin fibers, preferably the hair.

[0055] These properties can also be evaluated in-tubo by measuring the chelating activity of transition metals such as iron (Fe), for example with tryptophan degradation tests and / or tests of formation of dityrosine and / or pentosidine and / or fluorescent glycation products, measured in the presence of the hydrolyzed extract according to the invention, following the oxidation of albumin, used as a protein model, in the presence of hydrogen peroxide and iron as an oxidation catalyst.

[0056] In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the biomechanical properties of skin appendages when the protection index calculated under the conditions of Example 4 for the tryptophan degradation test is greater than 10%, advantageously greater than 15%, more advantageously greater than 20%, particularly greater than 25%, more particularly greater than 30%, and even more particularly greater than 35%. Advantageously, the skin appendages are keratin fibers, preferably hair.

[0057] In an alternative embodiment, the hydrolyzed extract according to the invention is in an effective quantity for "maintaining and / or increasing the biomechanical properties" of skin appendages when the percentage of dityrosine formation is decreased in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 4, by at least 10%, advantageously by at least 15%, more advantageously by at least 20%, preferably by at least 25% compared to the percentage of dityrosine formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index calculated under the conditions of Example 4 for the dityrosine formation test is greater than 30%, advantageously greater than 35%, more advantageously greater than 60%, in particular greater than 65%. Advantageously, the skin appendages are the keratin fibers, preferentially the hair.

[0058] In an alternative embodiment, the hydrolyzed extract according to the invention is, in sufficient quantity to "maintain and / or enhance the biomechanical properties" of skin appendages, when the percentage of pentosidine formation in the presence of the hydrolyzed extract according to the invention is reduced, preferably under the conditions described in Example 4, by at least 10%, advantageously by at least 25%, compared to the percentage of pentosidine formation measured in the absence of the extract. hydrolyzed according to the invention, and / or when the protection index calculated under the conditions of Example 4 for the pentosidine formation test is greater than 50%, advantageously greater than 60%. Advantageously, the skin appendages are the keratin fibers, preferably the hair.

[0059] In an alternative embodiment, the hydrolyzed extract according to the invention is effective in "maintaining and / or increasing the biomechanical properties" of skin appendages when the percentage of formation of fluorescent glycation products in the presence of the hydrolyzed extract according to the invention is reduced, preferably under the conditions described in Example 4, by at least 5%, advantageously by at least 10%, more advantageously by at least 13%, compared to the percentage of formation of glycation products measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index calculated under the conditions of Example 4 for the test of formation of fluorescent glycation products is greater than 10%, advantageously greater than 15%, more advantageously greater than 20%, in particular greater than 25%. Advantageously, the skin appendages are the keratin fibers, preferentially the hair.

[0060] The biomechanical properties according to the invention can also be evaluated in-tubo by measuring the chelating activity of transition metals such as copper (Cu), for example with the lipid oxidation test evaluated by the formation of ma-londialdehyde (MDA) in the presence of copper (Cu) (oxidation catalysts) and thiobarbituric acid (TBA) in the presence of the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in "maintaining and / or increasing the biomechanical properties" of skin appendages when the percentage of lipids oxidized by the MDA assay in the presence of the hydrolyzed extract according to the invention is reduced, preferably under the conditions described in Example 5, by at least 20%, advantageously by at least 40%, more advantageously by at least 60%, preferably by at least 75%, compared to the percentage of lipids oxidized by the MDA assay measured in the absence of the hydrolyzed extract according to the invention and / or when the protection index calculated under the conditions of Example 5 for the test of formation of fluorescent glycation products is greater than 60%, advantageously greater than 75%, more advantageously greater than 90%, in particular greater than 95%. Advantageously, skin appendages are keratin fibers, preferentially hair.

[0061] These properties can also be evaluated in vitro by measuring the anti-glycation activity on the proteins of skin appendages, for example with the dityrosine and / or pentosidine formation test measured after incubation of said appendages in a glucose solution in the presence of the hydrolyzed extract according to the invention. In an alternative embodiment, the hydrolyzed extract according to the invention is in effective quantity to "maintain and / or enhance the biomechanical properties" of skin appendages, when the percentage of dityrosine formation is reduced in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 6, by at least 5%, advantageously by at least 10%, more advantageously by at least 15%, compared to the percentage of dityrosine formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index calculated under the conditions of Example 6 for the dityrosine formation test is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, in particular greater than 55%. Advantageously, the skin appendages are the keratin fibers, preferably hair.

[0062] In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the biomechanical properties of skin appendages when the percentage of pentosidine formation in the presence of the hydrolyzed extract according to the invention is reduced, preferably under the conditions described in Example 6, by at least 5%, advantageously by at least 10%, more advantageously by at least 15%, compared to the percentage of pentosidine formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index calculated under the conditions of Example 6 for the pentosidine formation test is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and in particular greater than 55%. Advantageously, the skin appendages are keratin fibers, preferably hair.

[0063] Biomechanical properties can also be evaluated ex-vivo by measuring the carbonylation of keratins and their associated proteins, for example by the particle (pollution) and UV test, measured in the presence of the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the biomechanical properties of skin appendages when the percentage of carbonyl proteins (oxidation products) is reduced in the presence of the hydrolyzed extract according to the invention, particularly in the cuticle, and even more particularly in the cortex, preferably under the conditions described in Example 7, by at least 5%, advantageously by at least 15%, more advantageously by at least 20%, preferably by at least 25%, and more advantageously by at least 30% in the cortex, compared to the percentage of carbonyl proteins measured in the absence of the hydrolyzed extract according to the invention, and / or when the sun protection factor calculated under the conditions of Example 7 is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and in particular greater than 55%.Advantageously, skin appendages are keratin fibers, preferentially hair.

[0064] The biomechanical properties can also be evaluated in-vitro by measuring the oxidative stress of the skin appendages, for example with the optical measurement of the birefringence properties, in particular the birefringence index (Kindex) of the skin appendages treated with the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the biomechanical properties of skin appendages when the Kindex value in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 7c, increases by at least 5%, advantageously by at least 10%, more advantageously by at least 13%, compared to the Kindex value measured in the absence of the hydrolyzed extract according to the invention, and / or when the improvement index calculated under the conditions of Example 7c is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and in particular greater than 55%. Advantageously, the skin appendages are the keratin fibers, preferably hair. Further advantageously, this refers to the hydrolyzed extract as prepared according to Example 1a).

[0065] Biomechanical properties can also be evaluated ex vivo by measuring the hair protection score (SPC) to classify hair care products on damaged skin appendages (UVA irradiation and particles) treated with the hydrolyzed extract according to the invention. In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or increasing the biomechanical properties of the skin appendages, preferably under the conditions described in Example 7d, when the SPC is greater than 30, preferably equal to or greater than 50. Advantageously, the skin appendages are the keratin fibers, preferably the hair.

[0066] These properties can also be evaluated ex-vivo by measuring the stabilization of cysteine ​​groups, for example by measuring the vibrational spectra of keratins from damaged skin appendages treated with the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the biomechanical properties of skin appendages when the absorption bands of 1174, 1116, 1150, and / or 840 cm¹ of damaged skin appendages decrease in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 9, by at least 5%, advantageously by at least 15%, more advantageously by at least 20%, and even more advantageously by at least 35%, compared to the absorption bands of 1174, 1116, 1150, and / or 840 cm¹ measured in the absence of the hydrolyzed extract according to the invention on skin appendages damaged by bleaching, and / or when the efficacy index calculated under the conditions of Example 9 is greater than 5%, advantageously greater than 10%, plus. advantageously greater than 13%. Advantageously, the cutaneous annexes are the keratin fibers, preferentially the hair.

[0067] The biomechanical properties, in particular of strength and / or plasticity and / or resistance and / or flexibility, can also be evaluated ex-vivo by measuring the stability of the damaged skin appendages, for example by measuring the denaturation temperature of the proteins of the damaged skin appendages using hydrogen peroxide, and then treated with the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or increasing the strength, plasticity, resistance, and / or elasticity of skin appendages when the denaturation temperature increases in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 10, by at least 2°C, advantageously by at least 5°C, compared to the denaturation temperature measured in the absence of the hydrolyzed extract according to the invention, and / or when the repair index compared to skin appendages treated with placebo or untreated, calculated under the conditions of Example 10, is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and in particular greater than 60%. Advantageously, the skin appendages are keratin fibers, preferably hair.

[0068] These properties can also be evaluated by tensile strength tests (Diastron), fatigue tests, or repeated hair-styling tests to assess breakage. It is also possible to quantify hair fibers with split ends. Furthermore, these biomechanical properties can be evaluated by measuring the styling force required on dry or wet hair. In particular, these properties can be evaluated ex vivo by measuring the rupture of skin appendages, for example, by a fatigue test on damaged skin appendages, preferably treated with hydrogen peroxide and then treated with the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or increasing the strength, plasticity, resistance, and / or flexibility of skin appendages when the number of cycles before rupture of the skin appendages or the hair fiber strengthening index in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 11, is greater than 10%, preferably greater than 20%, and more preferably greater than 30%, compared to the number of cycles before rupture of the skin appendages or the strengthening index measured in the absence of treatment with the hydrolyzed extract according to the invention. Advantageously, the skin appendages are the keratin fibers, preferably the hair. Further advantageously, this refers to the hydrolyzed extract as prepared according to Example 1a).

[0069] In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or increasing the strength and / or plasticity and / or resistance and / or flexibility of skin appendages when the reduction in the percentage of breakage after repeated styling of damaged skin appendages, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 13, is greater than 50%, preferably greater than 70%, and more preferably greater than 75%, compared to the percentage of breakage of the skin appendages measured in the absence of treatment with the hydrolyzed extract according to the invention. Advantageously, the skin appendages are the keratin fibers, preferably the hair. Further advantageously, they are the hydrolyzed extract as prepared according to Example 1).

[0070] The biomechanical properties of color and / or brightness can be evaluated ex-vivo by measuring the color variations of the skin appendages, for example by a colorimetric test (Delta a* and Delta E*) on damaged skin appendages, preferably using hydrogen peroxide, and having undergone sun exposure, then treated with the hydrolyzed extract according to the invention.In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or enhancing the color and / or radiance of skin appendages when the Delta a* value in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 12, increases by 4%, preferably by 6%, compared to the Delta a* value measured in the absence of the hydrolyzed extract according to the invention and / or the % effectiveness calculated under the conditions of Example 12 for Delta a* is at least 20%, advantageously at least 30%, in particular at least 80% and / or the Delta E* value decreases by 2%, preferably by 3%, compared to the Delta E* value measured in the absence of treatment with the hydrolyzed extract according to the invention and / or the % effectiveness calculated under the conditions of Example 12 for Delta E* is of at least 15%, advantageously of at least 20%, in particular of at least 25%.Advantageously, skin appendages are keratin fibers, preferentially hair.

[0071] The biomechanical properties can also be evaluated in vitro by measuring the amount of ATP in the fibroblasts of the papilla of hair follicles. In an alternative embodiment, the hydrolyzed extract according to the invention is effective in maintaining and / or increasing the biomechanical properties of skin appendages, preferably keratin fibers, advantageously hair, when the percentage of papilla fibroblast aggregates in the presence of the hydrolyzed extract according to the invention, preferably in the presence of the hydrolyzed extract as prepared according to Example 1a, under the conditions described in Example 14, is greater than 10%, preferably greater than 20%, and more preferably is greater than 25%, advantageously greater than 30% compared to the percentage of fibroblast aggregates of the papilla measured in the absence of treatment with the hydrolyzed extract according to the invention.

[0072] The hydrolyzed extract according to the invention is therefore effective and can be used to maintain and / or increase the resistance and / or flexibility and / or strength and / or volume and / or color and / or shine and / or plasticity of skin appendages, and preferably those of the hair, and is thus able to reduce their fragility, breakage, roughness, and thus improve detangling and / or styling.

[0073] The term "maintaining and / or enhancing the biomechanical properties" of skin appendages, advantageously hair, also means making said skin appendages, advantageously hair, visually smoother and / or shinier and / or brighter and / or more manageable and / or less split. These properties can be evaluated using imaging techniques, for example, video microscopy or electron microscopy, or through sensory tests, for example, on hair strands, carried out by external volunteers trained to describe their visual or tactile sensations, or through evaluation tests by experts (hairdressers, for example) or trained volunteers, or through self-assessment via consumer questionnaires. The effectiveness of a product in its formula is evaluated according to perceptible visual or tactile quality criteria, particularly those related to hair.

[0074] The repair and / or enhancement of the biomechanical properties of the hydrolyzed extract according to the invention make it an active cosmetic ingredient for improving the resistance and / or strength and / or volume and / or color and / or shine and / or luster of skin appendages, in particular keratin fibers, preferably hair.

[0075] The enzymatic hydrolysis to obtain the hydrolyzed extract according to the invention can be carried out in the presence of any protease known to those skilled in the art, and advantageously in the presence of an enzyme of animal origin chosen from pepsin, pancreatic enzymes such as trypsin or chymotrypsin, preferably trypsin, an enzyme of plant origin chosen from papain, bromelain, ficin, actinidin, preferably papain, or an enzyme of bacterial origin chosen from the enzyme from the Bacillus licheniformis strain, in particular marketed under the name Alcalase®, or from the B. subtilis strain, preferably the enzyme from the Bacillus licheniformis strain marketed under the name Alcalase®.In a particularly advantageous embodiment of the invention, the enzyme used is derived from the Bacillus licheniformis strain and is marketed in particular under the name Alcalase®.

[0076] The quantity by weight of Silybum marianum seed cake according to the invention used for hydrolysis is advantageously between 1% and 20%, more advantageously between 2% and 15%, even more advantageously between 10% and 15%, by weight relative to the total weight of the solvent and the cake.

[0077] The solvent that can be used for hydrolysis can be an aqueous solvent.

[0078] According to the invention, the term "aqueous solvent" means an aqueous extraction solution containing more than 60% by weight, advantageously at least 70% by weight, in particular at least 80% by weight, more particularly at least 90% by weight, and especially at least 95% by weight, of water relative to the total weight of the aqueous solution, and even more advantageously not containing glycol and especially not containing alcohol. The aqueous solvent does not contain hydrotropic aqueous solution, subcritical water, ethanol, isopropanol, or propylene glycol.

[0079] Advantageously, the solvent used is water as the sole solvent.

[0080] Hydrolysis can be carried out at a pH between 3 and 10 depending on the optimum pH of the enzyme, and advantageously at a pH between 7 and 9, very advantageously at a pH between 7.5 and 8.5, even more advantageously at a pH of 7.5.

[0081] Alternatively, enzymatic hydrolysis is carried out in the presence of papain, at a pH between 3 and 5.5, preferably between 4.5 and 5.

[0082] Hydrolysis can be carried out at a temperature between 30°C and 75°C, advantageously between 50°C and 70°C, in particular between 55°C and 65°C, preferably at 65°C. It can be carried out for a period of 30 minutes to 24 hours, preferably from 45 minutes to 12 hours, more preferably from 50 minutes to 6 hours, most preferably from 1 to 3 hours.

[0083] The quantity by weight of enzyme used for hydrolysis is between 0.1% and 25% by weight, preferably between 0.5% and 20% by weight, more preferably between 0.75% and 10% by weight, advantageously between 1% and 5% by weight, relative to the weight of the cake.

[0084] The enzyme used can then be inactivated by heating, advantageously at a temperature of 80°C to 100°C and very advantageously at a temperature of 90°C, in particular for a period of 10 minutes to 3 hours, preferably from 10 minutes to 2 hours, more preferably from 20 minutes to 1.5 hours. The inactivation of the enzyme can take place at pH between 3.5 and 8, preferably between 4 and 7, more preferably between 4 and 6.5, advantageously at pH 6.0.

[0085] The hydrolyzed extract obtained can then be centrifuged and clarified by successive filtrations up to a porosity of 0.22pm (filtration threshold).

[0086] The hydrolyzed extract according to the invention can therefore be in liquid form advantageously comprising a dry matter content of 1% to 20% by weight, advantageously of 2% to 10% by weight, more advantageously between 3% and 6% by weight, relative to the total weight of the hydrolyzed extract in liquid form.

[0087] In an alternative embodiment, an aqueous extraction step is carried out before the hydrolysis step, by any method known to those skilled in the art, in particular by maceration, and advantageously by extraction with water as the sole solvent. The quantity by weight of press cake used for the aqueous extraction is between 1 and 20%, advantageously between 2% and 15%, and further advantageously between 10% and 15%, by weight relative to the total weight of solvent and press cake.

[0088] In this case, the aqueous extraction can be carried out at a pH between 7.5 and 12, advantageously at a pH between 8 and 9, very advantageously at a pH of 9, advantageously for a period of 30 minutes to 2 hours, preferably for a period of 1 hour.

[0089] Aqueous extraction can be carried out at a temperature between 4°C and 300°C, advantageously between 15°C and 100°C, even more advantageously between 20°C and 90°C, and most advantageously between 20°C and 70°C. In a particularly advantageous embodiment of the invention, aqueous extraction is carried out at ambient temperature, i.e., between 20 and 25°C.

[0090] In a preferred embodiment, the aqueous extraction is carried out at 20°C, at pH 12, from a quantity of 15% by weight of cake relative to the total weight of cake and water.

[0091] In another embodiment, the aqueous extraction is carried out at 20°C, at pH 12, from a quantity of 12.5% ​​by weight of cake relative to the total weight of cake and water.

[0092] In an alternative embodiment, the aqueous extraction is carried out under subcritical conditions.

[0093] Extraction under "subcritical conditions" refers to extraction in the presence of water, at temperatures above 100°C and pressures below 22.1 MPa (221 bar), such that the water remains in a liquid state but has a viscosity and surface tension lower than that of water at ambient temperature, thus increasing its dielectric constant. The extraction pressure will therefore be between 0.2 MPa (2 bar) and 22.1 MPa (221 bar), preferably between 1 and 5 MPa (10 and 50 bar), and more preferably between 1 and 2 MPa (10 and 20 bar).

[0094] Thus, under subcritical conditions, the extraction is carried out in water, in particular at a temperature ranging from 100°C to 300°C, advantageously from 120°C to 250°C, more advantageously between 120°C and 180°C. The extraction can be carried out at a single given temperature or at successive increasing temperatures. In an advantageous embodiment of the invention, the extraction will be carried out at a single temperature of 160°C. In an alternative mode, it will be conducted according to a gradient of three increasing temperatures between 100°C and 200°C, such as 120°C, 140°C then 160°C or 110°C, 130°C then 150°C, or even 120°C, 145°C then 170°C.

[0095] In a preferred embodiment, the subcritical extraction is carried out at a pressure of 5 MPa (50 bars), at a temperature of 120°C for 20 minutes. The extract obtained is then hot-filtered.

[0096] In an alternative embodiment, the subcritical extraction is carried out at a pressure of 5 MPa (50 bar), at a temperature of 140°C for 20 minutes. The extract obtained is then hot-filtered.

[0097] In an alternative embodiment, the hydrolyzed extract of Silybum marianum seed cake in liquid form thus obtained can be used as a solvent to hydrolyze a new quantity of Silybum marianum seed cake.

[0098] In an advantageous embodiment, the hydrolyzed extract is obtained by enzymatic digestion as follows: Silybum marianum seed cake is ground and suspended in water (as the sole solvent) at a concentration of 12.5% ​​by weight of seed cake relative to the total weight of cake and solvent. It is then hydrolyzed for 1 hour at a temperature of 55°C and pH 8.5, with the pH maintained during hydrolysis, using a 1% by weight concentration of a bacterial protease from the Bacillus licheniformis strain (Alcalase®) in liquid form relative to the weight of the seed cake. The enzyme is then inactivated by heating for 1 hour at a temperature of 90°C and pH 4.5. The mixture is then cooled, centrifuged, and the supernatant filtered (filtration threshold: 0.22 µm), under the conditions described in example 1b).

[0099] In a particularly advantageous embodiment of the invention, the hydrolyzed extract is obtained by enzymatic digestion as follows: the Silybum marianum seed cake is ground and suspended in water (as the sole solvent) at a concentration of 15% by weight of seed cake relative to the total weight of cake and solvent, and is subjected to hydrolysis for a period of 2 hours at a temperature of 65°C and pH 7.5, with the pH maintained during hydrolysis, with a concentration by weight of 1.7% of bacterial protease from the Bacillus licheniformis strain (Alcalase®) in liquid form relative to the weight of the seed cake. The enzyme is then inactivated by heating for 1 hour at a temperature of 90°C and pH 6.0. The mixture is cooled to room temperature (between 20°C and 30°C), centrifuged and filtered (filtration threshold: 0.22 pm) under the conditions as described in example 1a).

[0100] The terms "small peptides" or "low molecular weight peptides" mean peptides having a weight-average molecular weight of less than 7000 Daltons, analyzed by gel permeation on a column (of the Superdex® peptide 10 / 30 HR GE type or the Superose® 12 10 / 300 GL type) followed by calibration of the column with molecules proteins of known molecular weights (gel permeation chromatography (GPC)).

[0101] Advantageously the hydrolyzed extract according to the invention has a total protein dry mass content of between 50% and 95% by weight, preferably between 60% and 90% by weight, more preferably between 60% and 70% by weight, relative to the total dry mass weight of the hydrolyzed extract according to the invention.

[0102] Advantageously, the hydrolyzed extract according to the invention has a dry mass content of peptides with an average molecular weight by weight of between 100 Da and 20kDa, analyzed by column gel permeation (of the Superose® 12 10 / 300 GL type), of between 50% and 90%, preferably between 65% and 90%, more preferably between 75% and 90%, by weight relative to the total dry mass weight of the hydrolyzed peptides of the hydrolyzed extract according to the invention.

[0103] Advantageously, the hydrolyzed extract according to the invention has a dry mass content of peptides with average molecular weights by weight between 1OODa and 700 Da, analyzed by gel permeation on a Superdex® peptide 10 / 30 HR GE column, of between 60% and 90%, advantageously between 70% and 80%, more advantageously between 75% and 80%, by weight relative to the total dry mass weight of peptides with molecular weights between 1OODa and 7000 Da of the hydrolyzed extract according to the invention (column sensitivity).

[0104] Advantageously, the hydrolyzed extract according to the invention has a dry mass content of peptides with average molecular weights by weight between 1OODa and 2500 Da, analyzed by gel permeation on a Superdex® peptide 10 / 30 HR GE column, of between 70% and 99%, advantageously between 80% and 99%, more advantageously between 90% and 99%, by weight relative to the total dry mass weight of peptides with molecular weights between 1OODa and 7000 Da of the hydrolyzed extract according to the invention (column sensitivity).

[0105] Even more advantageously, the hydrolyzed extract according to the invention is not used in combination with a retinoid.

[0106] The hydrolyzed extract according to the invention can be used alone or incorporated into a cosmetic composition.

[0107] In one embodiment, the hydrolyzed extract can then be dried, for example, by freeze-drying or spray-drying, with or without maltodextrin. The hydrolyzed extract is then in powder form.

[0108] According to one embodiment, the hydrolyzed extract according to the invention, in particular obtained under the conditions described in examples 1a) and 1b), can be atomized in the presence of a maltodextrin concentration by weight of between 1% and 99%, advantageously between 5% and 90%, preferably between 40% and 85%, even more preferably between 65% and 85%, and more preferably between 70% and 80% relative to to the total weight of the powder obtained and of a concentration by weight of hydrolyzed extract according to the invention of between 1% and 99%, advantageously between 10% and 95%, preferably between 15% and 60%, even more preferably between 15% and 25%, more preferably between 20% and 30% relative to the total weight of the powder obtained, advantageously under the conditions of examples i) or id).

[0109] Alternatively, the hydrolyzed extract can be used alone in solid form, as a cosmetic or dermatological ingredient.

[0110] Alternatively, when used alone in liquid form, as a cosmetic or dermatological ingredient, it is advantageously solubilized in an aqueous solution containing glycerin, advantageously present at a concentration of 60% to 90%, further advantageously of 70% to 85%, very advantageously at a concentration of 80%, by weight relative to the total weight of the aqueous solution comprising the hydrolyzed extract.

[0111] In an alternative embodiment of the invention, the hydrolyzed extract shall be solubilized and / or diluted in a solvent, in particular a polar solvent, such as water, an alcohol, a polyol, a glycol, such as pentylene glycol and / or butylene glycol and / or hexylene glycol and / or caprylyl glycol, or a mixture thereof, preferably a hydroglycolic mixture, more preferably containing a glycol selected from hexylene glycol, propylene glycol, caprylyl glycol and any mixture thereof. Advantageously, the hydrolyzed extract according to the invention is diluted and / or soluble in an aqueous solution containing hexylene glycol, in particular containing between 0.1% and 10% by weight of hexylene glycol, preferably between 0.5% and 5% by weight of hexylene glycol, relative to the total weight of the cosmetic ingredient.Advantageously, the hydrolyzed extract according to the invention is diluted and / or soluble in an aqueous solution containing caprylyl glycol, in particular containing between 0.01% and 5% by weight of caprylyl glycol, preferably between 0.1% and 1% by weight of caprylyl glycol, relative to the total weight of the aqueous solution comprising the hydrolyzed extract. Alternatively, the solution in which the hydrolyzed extract according to the invention is solubilized comprises pentylene glycol and caprylyl glycol.

[0112] In particular, the aqueous solution in which the hydrolyzed extract according to the invention is solubilized comprises xanthan gum, in particular between 0.01% and 5% by weight of xanthan gum relative to the total weight of the aqueous solution, more particularly between 0.1% and 1% by weight of xanthan gum relative to the total weight of the aqueous solution comprising the hydrolyzed extract.

[0113] The hydrolyzed extract can be used alone as a cosmetic ingredient or incorporated into a cosmetic composition comprising at least one cosmetically acceptable excipient.

[0114] For the purposes of the present invention, the term "cosmetically acceptable" excipient means a cosmetically acceptable compound and / or solvent, in particular topically acceptable, i.e., not inducing an undue inflammatory and allergic response on contact with the skin, in particular the scalp, non-toxic, non-unstable, or their equivalents.

[0115] For the purposes of this invention, "cosmetic composition" means a non-therapeutic composition, that is, a composition intended for the prevention and / or care of the skin, particularly the scalp, and / or skin appendages, particularly hair, which a dermatologist would describe as "normal," that is, non-pathological. "Normal" skin, scalp, skin appendage, or hair means healthy skin, scalp, skin appendage, or hair as defined above.

[0116] In a preferred embodiment of the invention, the hydrolyzed extract according to the invention is present in the cosmetic composition in a content of between 1x10 4% to 10% by weight, preferably from 1x10 3% to 5% by weight, more preferably from 1x10 3% to 3% by weight, even more preferably from 1x10 3% to 1% by weight, advantageously from 0.01 to 1% by weight, relative to the total weight of the composition.

[0117] The composition can therefore be used to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

[0118] The cosmetic composition according to the invention can be presented in the galenic forms classically used for topical application on the skin or scalp and / or skin appendages, preferably the scalp and / or skin appendages, such as liquid or solid forms or even in the form of liquid under pressure.They may be formulated as a solution, aqueous or oily, an aqueous cream or gel or an oily gel, in particular in a jar or tube, in particular a shower gel, a shampoo, a conditioner, a milk, an oil, an emulsion, a hydrogel, a microemulsion or a nanoemulsion, in particular oil-in-water or water-in-oil or multiple or silicone, a serum, a lotion, in particular in a glass bottle, a plastic bottle or a dosing bottle or in an aerosol or spray, an ampoule, a liquid soap, a paste, a dermatological bar, an ointment, a mousse, a mask, a lacquer, a patch, a varnish, an anhydrous product, preferably liquid, pasty or solid, for example in the form of a stick or in powder form.This could be a makeup product, specifically for eyelashes or eyebrows such as mascara or pencil, or a makeup remover for eye care and beauty, and / or a product for nail care and beauty, and / or for beard care and / or beauty. In particular, the cosmetic composition is chosen from [specific criteria / details]. the group consisting of a serum, a lotion, a cream, a shampoo, a conditioner, an oil, a milk, an ointment, a paste, a mousse, an emulsion, a hydrogel, a shower gel, a mask, a lacquer, a spray, a wax, a mascara, a makeup pencil, a varnish, advantageously a shampoo, a conditioner or a lotion.

[0119] Preferably, the hydrolyzed extract according to the invention is suitable for formulations with a neutral and gentle composition that respects skin appendages, advantageously keratin fibers, particularly hair fibers. The hydrolyzed extract according to the invention is also suitable for use in cationic formulations with surfactants.

[0120] The cosmetic compositions according to the invention may contain any suitable solvent and / or any suitable vehicle and / or any suitable excipient, optionally in combination with other compounds of interest. In particular, they may contain a cosmetically acceptable excipient selected from surfactants, preservatives, buffering agents, swelling agents, chelating agents, biocidal agents, denaturants, opacifying agents, pH adjusters, reducing agents, stabilizing agents, emulsifiers, thickeners, gelling agents, film-forming polymers, solvents, fillers, bactericides, odor absorbers, mattifying agents, conditioning agents, texturizing agents, gloss-enhancing agents, pigments, colorants, perfumes, and chemical or mineral sunscreens, trace elements, and essential oils. These combinations are also covered by the present invention.

[0121] The cosmetic composition may contain other cosmetic agents having properties identical to the hydrolyzed extract according to the invention and inducing a synergistic or non-synergistic effect with said hydrolyzed extract, or may contain cosmetic agents with complementary effects such as anti-hair loss agents, hair protectants, soothing agents, anti-aging agents, or anti-pollution agents. Examples of anti-hair loss agents include the combination of sulfopeptides, amino acids, aminosaccharides, B vitamins, zinc, and extracts of Panax ginseng and Artium majus marketed under the name Trichogen™ LS 8960 by the applicant, or a hair protectant such as an extract of the pericarp of Litchi chinensis marketed under the name Litchiderm™ by the applicant, or a soothing and anti-itch agent such as rapeseed phytosterols marketed under the name Phytosoothe™ LS9766 by the applicant.

[0122] Other active ingredients may be present in the composition, such as an extract of Cassia alata leaves marketed under the name DN-Age™ as an antioxidant active ingredient for hair care in particular, a combination of an extract of Salvia miltiorhizza and niacinamide marketed under the name CollRepair™ as a deglycating agent, or active ingredients promoting skin and therefore scalp firmness hair-related products, such as a synthetic tetrapeptide marketed under the name Dermican™, an extract of Hibiscus abelmoschus marketed under the name Li-nefactor™, a purified pea extract marketed under the name Proteasyl™, an extract of Manilkara multinervis marketed under the name Elestan™, an extract of Khaya senegalensis marketed under the name Collalift™18, an Argan pulp extract marketed under the name Argassential™ by the applicant, an extract of Schizandra chinensis marketed under the name Sqisandryl™, an extract of Eperuafalcata marketed under the name Eperuline™, or an extract of Orthosiphon staminus marketed under the name MAT-XS™ Bright, all marketed by the applicant. These combinations of active ingredients help to strengthen the hair follicle and reduce hair loss.The hydrolyzed extract of the invention can also be combined with a seed extract of the plant Nephelium lappaceum, marketed under the name Rambuvital™ by the applicant for its hair-protective properties, particularly against pollution. The hydrolyzed extract can also be used in combination with a hydrolyzed extract of Hippophae rhamnoides seed cake.

[0123] Advantageously the cosmetic composition according to the invention does not comprise a retinoid.

[0124] A third object of the invention relates to a cosmetic, non-therapeutic care method, comprising the topical application of the hydrolyzed extract according to the invention or of a cosmetic composition comprising it, to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

[0125] In an advantageous embodiment of the invention, the cosmetic care process consists of the topical application of the hydrolyzed extract according to the invention or of the cosmetic composition comprising it to all or part of the skin of the body and / or face including skin appendages, in particular chosen from the scalp, legs, thighs, arms, belly, décolleté, neck, all or part of the face, forehead, chin, lip contour, eye contour, the so-called "T-shaped" area of ​​the face, and advantageously the scalp, and / or to all or part of the skin appendages, advantageously on the nails, hair, body hair, in particular beard hair, eyelashes and / or eyebrows, even more advantageously on the hair, preferably on all or part of the keratin fibers, more preferably on all or part of the hair.

[0126] The cosmetic care process therefore makes it possible to maintain and / or increase the resistance and / or strength and / or volume and / or color and / or shine and / or plasticity, i.e., the non-brittle appearance, and / or the suppleness of the skin appendages, advantageously of keratin fibers, more advantageously of hair.

[0127] Another object relates to a cosmetic treatment method for maintaining and / or increasing the biomechanical properties of skin appendages, preferably keratin fibers, advantageously hair, and comprising the steps of: - Identification on a human being not suffering from a pathology, in particular cutaneous, requiring therapeutic treatment, of an area of ​​skin of the body and / or face comprising cutaneous annexes and / or of an area of ​​cutaneous annexes for which it is desired to maintain and / or increase the biomechanical properties of said cutaneous annexes, preferably keratin fibers, advantageously hair, and / or which need it; - Topical application to said area of ​​the skin of the body and / or face comprising skin appendages and / or to said area of ​​skin appendages of the hydrolyzed extract according to the invention or of the composition comprising it, in particular at a content of between 1x10 4% to 10% by weight, preferably of 1x10 3% to 5% by weight, more preferably of 1x10 3% to 3% by weight, even more preferably of 1x10 3% to 1% by weight, advantageously of 0.01% to 1% by weight, relative to the total weight of the composition.

[0128] The examples form an integral part of the invention, and any feature that appears new compared to a prior art, taken from the description as a whole, including the examples, forms an integral part of the invention. Thus, each example has general application.

[0129] Unless otherwise stated, temperature is expressed in degrees Celsius and pressure is atmospheric pressure. Description of the implementation methods EXAMPLES

[0130] Example 1: Different methods of preparing the hydrolyzed extract of Silybum marianum seed cake according to the invention

[0131] The oilcake used in the following examples contains less than 8% by weight of lipids relative to the total weight of the oilcake.

[0132] Example a: Enzymatic hydrolysis - Hydrolyzed extract 1

[0133] A seed cake of Silybum marianum is ground and suspended in water (as the sole solvent) at a concentration of 15% by weight of seed cake relative to the total weight of seed cake and solvent is hydrolyzed for a period of 2 hours at a temperature of 65°C and pH 7.5, with the pH maintained during hydrolysis, with a weight concentration of 1.7% of bacterial protease from the Bacillus licheniformis strain (Alcalase®) in liquid form relative to the weight of the cake. The enzyme is then inactivated by heating for 1 hour at a temperature of 90°C and pH 6.0. The mixture is cooled to room temperature, centrifuged, and filtered (filtration threshold: 0.22 µm). A liquid product is obtained with a dry matter content of 4.14 ±0.3% by weight of the liquid product.

[0134] Example 1b: Enzymatic hydrolysis - Hydrolyzed extract 2

[0135] Silybum marianum seed cake is ground and suspended in water (as the sole solvent) at a concentration of 12.5% ​​by weight of seed cake relative to the total weight of cake and solvent. It is then hydrolyzed for 1 hour at 55°C and pH 8.5, with the pH maintained during hydrolysis, using a 1% liquid bacterial protease derived from the Bacillus licheniformis strain (Alcalase®) relative to the weight of the seed cake. The enzyme is then inactivated by heating for 1 hour at 90°C and pH 4.5. The mixture is then cooled, centrifuged, and the supernatant filtered (filtration limit: 0.22 µm). We obtain a liquid product having a dry matter content of 3.26% by weight relative to the weight of the liquid product.

[0136] Example: Hydrolyzed extract 1 as a cosmetic ingredient (with maltodextrin)

[0137] The extract obtained in Example 1a) is atomized in the presence of maltodextrin, to a final quantity of maltodextrin of 75% by weight relative to the total weight of the final atomized extract. The extract is thus obtained in powder form.

[0138] Example Id: Hydrolyzed extract 2 as a cosmetic ingredient (with maltodextrin)

[0139] The extract obtained in Example 1b) is atomized in the presence of maltodextrin, to a final quantity of maltodextrin of 75% (w / w) by weight relative to the total weight of the final atomized extract. The extract is thus obtained in powder form.

[0140] Example: Enzymatic hydrolysis using hydrolyzed extract 1 according to Example

[0141] Silybum marianum seed cake is ground and suspended in the hydrolyzed extract (sole solvent) obtained in example a) at a content of 15% by weight of cake relative to the total weight of cake and solvent. The resulting mixture is then hydrolyzed for 2 hours at a temperature of 65°C and pH 7.5, with the pH maintained during hydrolysis, with a weight concentration of 1.7% bacterial protein from the Bacillus licheniformis strain (Alcalase®) in liquid form relative to the weight of cake. The enzyme is then inactivated by heating for 1 hour at a temperature of 90°C and pH 6.0. The mixture is cooled to room temperature, centrifuged, and filtered (filtration threshold: 0.22pm). We obtain a liquid product having a dry matter content of 6.1% by weight relative to the weight of the liquid product.

[0142] Example 2, Peptide equivalent content of the hydrochloric acid extract according to Example 1 and as a cosmetic ingredient according to Example 2 and analysis of the peptide molecular weight profile

[0143] a) Total protein assay

[0144] The aim of this study is to quantify the total peptide equivalent content of the hydrolyzed extract according to Example 1 according to the invention.

[0145] Materials and methods: The peptide equivalent content of the hydrolyzed extract is estimated by determining the total nitrogen (Kjeldahl method - AOAC, Association of Official Analytical Chemists Official Methods of Analysis, AOAC, Washington, DC, USA, 2000) and multiplying the value obtained by a factor of 6.25 (N x 6.25). The results obtained are summarized in Table 1.

[0146] [Tables 1] Peptide equivalent content % by weight (Total nitrogen calculated for 100% dry mass of hydrolyzed extract according to Example la) Mean 63.1% Standard deviation 3.1%

[0147] Results: The amount of dry matter obtained after extraction and hydrolysis is between 3.84% and 4.44%. Total peptides represent between approximately 60% and 66.2% of the dry matter of the extract hydrolyzed according to example (Table 1).

[0148] Conclusion: The hydrolyzed extracts of Silybum marianum seed cakes according to the invention are rich in plant peptides (between 60% and 66.2% by weight of the total dry weight).

[0149] b) Analysis of the average molecular weight profile of hydrolyzed peptides

[0150] Molecular profile of the hydrolyzed extract

[0151] Objective: To characterize the average molecular weights by weight of the peptide compounds of the hydrolyzed extract according to the invention, 2 hydrolyzed extracts according to example la were studied and then classified into 3 categories: less than 100 Da, from 100 Da to 20 kDa (inclusive limits) and greater than 20 kDa.

[0152] Materials and methods: The weight-average molecular weight distribution of peptides from the hydrolyzed extract as a cosmetic ingredient, as described in example, is analyzed by gel permeation on a Superose® 12 10 / 300 GL column (GE Healthcare Life Sciences). The weight-average molecular weights are determined after calibrating the column with protein molecules of specified molecular weight. known average weight. The results obtained are summarized in Table 2.

[0153] [Tables2] Distribution by average molecular weight category of peptides contained in the extract according to Example (% by weight relative to the total weight of peptides) MW > 20,000 Da 20,000 Da < MW < 100 Da MW < 100 Da Mean 7.9 73.7 18.4 Standard deviation 1.5 2.5 1.1

[0154] Results: The peptides of the hydrolyzed extracts of Silybum marianum seed cake according to the invention are predominantly peptide compounds having an average molecular weight by weight between 100 Da and 20 kDa (Table 2).

[0155] Molecular profile between 100 Da and 7000 Da

[0156] Objective: The objective of this study is to investigate the distribution profile of the average molecular weights of peptides contained in hydrolyzed extracts of Silybum marianum seed cakes obtained according to Example 1, in the range between 100 Da and 7000 Da. The average molecular weights were then classified into 3 categories: the two extreme categories grouping the smallest molecular weights 100 Da < MW < 700 Da, and the largest molecular weights 2500 Da < MW < 7000 Da.

[0157] Materials and Methods: The weight-average molecular weight distribution of peptides from the hydrolyzed extract of Silybum marianum seed cake obtained according to Example 1 was analyzed by gel permeation on a Superdex® peptide 10 / 30 HR GE (GE Healthcare Life Sciences) column. Fractionation was performed from 100 Da to 7000 Da, and UV detection was carried out at 214 nm. The weight-average molecular weights were determined after calibrating the column with protein molecules of known weight-average molecular weight. The results obtained are summarized in Table 3.

[0158] [Tables3] Distribution of peptides in the lOODa range at 7000 Da according to average molecular weight (% by weight relative to the total weight of peptides in this range) Molecular Weight Ranges: 100 Da < Molecular Weight < 700 Da; 700 Da < Molecular Weight < 2500 Da; 2500 Da < Molecular Weight < 7000 Da. Hydrolyzed extract as a cosmetic ingredient, according to example: Average 77.15; 20.88; 1.97. Standard deviation 2.27; 2.40; 0.99.

[0159] Results: The hydrolyzed extracts of Silybum marianum seed cake according to the invention predominantly (98.03%) contain protein compounds of small average molecular weight by weight, i.e. in the range of 100 Da - 2500 Da (Table 3).

[0160] Example 3: In tubo effects of the hydrochloric acid extract according to Example 1b on the biomechanical properties of skin appendages - Anti-radical activity (DPPH)

[0161] The aim of this study is to measure the anti-radical activity of hydrolyzed extracts according to Example 1b by the DPPH method.

[0162] Materials and methods:

[0163] Diphenyl-1-picrylhydrazyl (DPPH°) is an oxidant that exists as a free radical lacking one electron. In this form, the dark purple compound exhibits absorption in the wavelengths between 510 nm and 530 nm. In the presence of an electron-donating reducing compound, diphenyl-2-picrylhydrazyl will form a stable compound accompanied by a decrease in its absorption properties (decolorization of the compound) (Blois M. Antioxidant Determinations by the Use of a Stable Free Radical. Nature 181, 1199-1200 (1958)). This method is used to select plant extracts with anti-radical properties (Kim BJ, Kim JH, Kim HP, Heo MY. Biological screening of 100 plant extracts for cosmetic use (II): anti-oxidative activity and free radical scavenging activity. Int J Cosmet Sci. 1997 Dec; 19(6):299-307).

[0164] The hydrolyzed extract of Silybum marianum seed cake according to Example 1b is mixed at 5% (w / v) in distilled water. 100 µl of this solution was tested in the presence of a similar volume of a 90 pM diphenyl-1-picrylhydrazyl solution (DPPH°, Sigma - 3.55 mg of DPPH powder in 100 mL of ethanol). After 30 minutes After incubation at room temperature and protected from light, absorbance was recorded at 530 nm using a spectrometer (Victor V, Perkin Elmer). The results are expressed as a percentage of DPPH° free radical inhibition relative to the negative control.

[0165] Results: They are summarized in Table 4. The hydrolyzed extract of Silybum marianum seed cakes according to the invention preserved the oxidation of DPPH up to 95% (Table 4).

[0166] [Tables4] DPPH Oxidation Mean (%) AND (%) % Oxidation Inhibition Hydrolyzed Extract according to Example 1b at 0.03% by weight 25.14 0.06 74.9 Hydrolyzed Extract according to Example 1b at 0.05% by weight 5.3 0.013 94.7

[0167] Conclusion: The hydrolyzed extract of Silybum marianum seed cakes according to the invention exhibits strong anti-radical activity and therefore an effect of maintaining and / or increasing biomechanical properties.

[0168] Example 4: In tubo effects of the hydrolyzed seed cake extract according to the invention on the biomechanical properties of skin appendages - Antioxidant / transition metal (Pc) chelating activity

[0169] Glycation reactions can occur chemically between reducing sugars and proteins. The presence of oxidizing agents and metals catalyzes this reaction and the formation of advanced glycation end products (AGEs).

[0170] The chelating and antioxidant activity of the hydrolyzed Silybum marianum press cake extract according to the invention was evaluated in an in tubo model in the presence of albumin and iron and an oxidizing agent, hydrogen peroxide. The oxidation reaction is assessed by measuring tryptophan degradation, tyrosine cross-linking, and the formation of glycation products, including pentosidine.

[0171] Materials and methods:

[0172] A reagent comprising 1.5% (w / v) albumin, 2 mM iron, 5 mM EDTA, and 25 mM hydrogen peroxide (H2O2) is mixed or not with the hydrolyzed extract of Silybum marianum press cake, as described in example 1a, at a concentration of 0.003% by weight or 0.01% by weight. Albumin oxidizes in the presence of H₂O₂. The oxidation reaction is catalyzed by iron. The catalysis of the oxidation reaction is checked in the absence of iron. The mixture is incubated at 37°C for 1 day.

[0173] The respective amounts of tryptophan, dityrosine, pentosidine and fluorescent glycation products are measured by fluorescence with the Thermo Scientific Varioskan® Flash device (excitation / emission wavelengths: 280 nm / 340 nm for tryptophan; 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosidine; 370 nm / 440 nm for terminal glycation products).

[0174] The threshold for statistical significance was set at 5% (p<0.05).

[0175] The protection index is calculated according to the following formula: (Control with Iron - Treatment with hydrolyzed extract according to the invention and with Iron) / (control with iron -control without iron) xlOO.

[0176] The test is carried out on the extract according to the example, dried so as to obtain a dry extract content of 100%.

[0177] [Tables5] Iron-catalyzed tryptophan degradation - level of undegraded tryptophan. Control: albumin without iron. Control: albumin with iron. Albumin with iron + hydrolyzed extract according to example 1a at 0.003% by weight. Albumin with iron + hydrolyzed extract according to example 1a at 0.01% by weight. Mean (%) 175 100 118 128. SD (%) 6 4 2 3. Difference (%) from control: albumin + iron 75 - 18 28. Protection index (%) - - 24 38. Statistics vs. control: albumin with iron p-value <0.001 (***•) - <0.001 (***) <0.001 (***)

[0178] The presence of iron catalyzes the oxidation reaction, observable by the increase in the level of undegraded tryptophan in the iron-free albumin control (Table 5). In the presence of the hydrolyzed extract of Silybum marianum seed cake according to example 1a at 0.003% and 0.01% (by weight), the degradation of tryptophan is significantly reduced with a recovery of 18% and 28% of residues, respectively. tryptophan (undegraded). The protection index of the hydrolyzed extract of Silybum marianum seed cake tested at 0.01% by weight is 38%.

[0179] [Tableauxô] Iron-catalyzed dityrosin formation Control Albumin e without Iron Control Albumin e with iron Albumin with iron + hydrolyzed extract of Silybum marianum cake according to example la at 0.003% by weight Albumin with iron + hydrolyzed extract of Silybum marianum cake according to example la at 0.01% by weight Mean (%) 58 100 84 72 SD (%) 2 12 6 2 Difference (%) - - -16 -28 Protection index (%) - - 38 67 Statistics vs control albumin with iron p-value <0.01 (**) - <0.05 (*) <0.01 (**)

[0180] The presence of iron catalyzes the formation of dityrosine in albumin in the presence of H2O2 (Table 6). The hydrolyzed extract of Silybum marianum seed cake, according to example 1a, at 0.003% by weight and 0.01% by weight, reduces dityrosine formation by 16% and 28%, respectively. The protection index of the hydrolyzed extract of Silybum marianum seed cake tested at 0.01% by weight is 67%.

[0181] [Tables?] Iron-catalyzed Pentosidine Formation Control albumin without iron Control albumin with iron Albumin with iron + hydrolyzed extract of Silybum marianum cake according to example la at 0.01% by weight Mean (%) 56 100 73 SD (%) 3 8 4 Difference (%) -44 - -27 Protection index (%) - - 61 Statistics vs control albumin with iron p-value <0.001 (***) - <0.001 (***)

[0182] The presence of iron also catalyzes the formation of pentosidine in the presence of H2O2 (Table 7). The hydrolyzed extract of Silybum marianum seed cake according to Example 1a at 0.01 wt% reduces pentosidine formation by 27%. The protection index of the hydrolyzed extract of Silybum marianum seed cake tested at 0.01 wt% is 61%.

[0183] [Tables8] Formation of fluorescent glycation products catalyzed by iron. Control: albumin without iron. Control: albumin with iron. Albumin with iron + hydrolyzed extract of Silybum marianum cake according to example la at 0.003% by weight. Albumin with iron + hydrolyzed extract of Silybum marianum cake according to example la at 0.01% by weight. Mean (%) 49 100 91 86 SD (%) 2 6 6 5 Difference (%) -51 - -9 -14 Protection index (%) - - 18 27 Statistics vs control: albumin with iron p-value <0.05 (*) <0.01 (**)

[0184] The presence of iron catalyzes the oxidation of albumin in the presence of H₂O₂ and is accompanied by the formation of fluorescent glycation products (Table 8). The hydrolyzed extract of Silybum marianum press cake, according to example 1a, at 0.003 wt% and 0.01 wt%, reduces the formation of these glycation products by 9% and 14%, respectively. The protection index of the hydrolyzed extract of Silybum marianum press cake tested at 0.003 wt% and 0.01 wt% is 18% and 27%, respectively.

[0185] Conclusion: The hydrolyzed extract of Silybum marianum seed cake reduces iron-catalyzed protein oxidation and the formation of glycation products and therefore has an effect on maintaining biomechanical properties.

[0186] Example 5: In tubo effects of the hydrolyzed seed cake extract according to the invention on the biomechanical properties of skin appendages - Antioxidant / transition metal chelating activity (Cul

[0187] The aim of this study is to evaluate the antioxidant / metal-chelating activity of the hydrolyzed extract of Silybum marianum cake according to the invention in an in tubo test, in the presence of lipoproteins and copper.

[0188] Materials and methods

[0189] Copper at 200 pM is contacted with low-density lipoproteins (LDL) at 100 pg / ml in a phosphate buffer saline solution. The extract Hydrolyzed Silybum marianum seed cake, as described in the example, at 0.048% by weight, is added to the solution or not. The reaction mixture is incubated at 37°C for 48 hours.

[0190] The level of lipid oxidation is assessed by quantifying malondialdehyde (MDA) after the addition of 2% (v) thiobarbituric acid (TBA) relative to the total volume of the reaction mixture. The fluorescence level is quantified spectrophotometrically using the Thermo Scientific Varioskan® Flash instrument, at an emission wavelength of 560 nm after excitation at a wavelength of 532 nm.

[0191] The Schiff base content is evaluated by spectrometry (excitation wavelength 370 nm / emission 440 nm) using the Thermo Scientific Varioskan® Flash instrument.

[0192] The threshold for statistical significance was set at 5% (p<0.05).

[0193] The protection index is calculated according to the following formula: (Treatment with hydrolyzed extract according to the invention and with LDL+Copper - LDL+Copper control) / (control without copper - LDL+copper control) xlOO.

[0194] Results: The results are summarized in Table 9 below.

[0195] [Tables9] Lipid content oxidized by malondialdehyde assay Control without Cu Control LDL + Cu LDL + Cu + hydrolyzed extract of Silybum marianum cake according to example la at 0.048% by weight Mean (%) 20 100 22 SD (%) 2 2 3 Difference (%) 80 - -78 Protection index (%) - - 97.5 Statistic vs control (t-test) p-value <0.001 (***•) - <0.001 (***)

[0196] The presence of copper potentiated the oxidation of lipids as measured by malondialdehyde. The hydrolyzed extract of Silybum marianum seed cake according to example la, used at 0.048 wt%, significantly reduced malondialdehyde formation by 78% (Table 9). The protection index of the hydrolyzed extract of Silybum marianum seed cake according to example la, tested at 0.048 wt%, is 97.5%.

[0197] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the invention reduced copper-catalyzed lipid oxidation and thus enabled maintain and / or increase biomechanical properties.

[0198] Example 6: In vitro effects of the hydrolyzed seed cake extract according to the invention on the biomechanical properties of skin appendages - Anti-freezing activity on hair fiber proteins

[0199] Objective: The aim of this study is to measure the anti-glycation effect of the hydrolyzed extract of Silybum marianum seed cake according to Example 1a on hair fiber proteins incubated in a glucose solution.

[0200] Materials and methods:

[0201] Standardized and cleaned hair strands were incubated in a 1 mol.L⁻¹ (M) glucose phosphate buffer solution, with or without the hydrolyzed extract as per example, and dried to obtain a 100% dry extract with a content of 0.01% by weight, at pH 7.4 for 2 days at 50°C. The strands were then rinsed, wrung out, combed, separated, and cut into 1 cm lengths. Proteins were extracted in a sodium hydroxide (NaOH) solution of ¹⁰N for 24 hours at room temperature, with gentle stirring.

[0202] The amount of dityrosine and pentosidine (glycation products) is measured by fluorescence with the Thermo Scientific Varioskan® Flash device (excitation / emission wavelengths: 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosidine).

[0203] The protection index is calculated according to the following formula: (Glycated control -Glycated control +Treatment with the extract according to the invention) / (non-glycated control -glycated control) xlOO.

[0204] The threshold for statistical significance was set at 5% (p<0.05).

[0205] Results: They are presented in Tables 10 and 11 below.

[0206] [TableauxlO] Dityrosine content on glycated hair (in the presence of glucose) Control unglycated hair Control glycated hair Glycated hair + Hydrolyzed extract of Silybum marianum seed cake according to example la at 0.01% by weight Mean (%) 72 100 84 SD (%) 9 11 6 Difference (%) 28 - -16 Protection index (%) - - 57 Statistic vs control (t-test or Mann-Whitney) p-value <0.001 (***) - <0.05 (*)

[0207] Incubating the fibers in a glucose solution increased the dityrosine content in the hair proteins. The hydrolyzed extract of Silybum marianum seed cake, according to example la, used at 0.01 wt%, significantly decreased dityrosine formation by 16% (Table 10). The protection index of the hydrolyzed extract, according to example la, used at 0.01 wt% against dityrosine production is 57%.

[0208] [Tables] Pentosidine content on glycated hair (in the presence of glucose) Control: unglycated hair Control: glycated hair Glycated hair + hydrolyzed extract of Silybum marianum seed cake according to example (la) at 0.04% by weight Mean (%) 71 100 83 SD (%) 6 9 10 Difference (%) 29 - -17 Protection index (%) - - 59 Statistic vs control (t-test) p-value <0.001 (***•) - <0.01 (**)

[0209] Incubating the fibers in a glucose solution increased the pentosidine content in the hair proteins. The hydrolyzed extract of Silybum marianum seed cake, according to the example used, at 0.04% by weight, decreased the amount of pentosidine in the hair proteins. The formation of pentosidine was significant at 17% (Table 11). The protection index of the hydrolyzed extract used at 0.04% by weight against the production of dityrosine was 59%.

[0210] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the invention reduced the formation of glycation products in the presence of glucose in the hair fiber and therefore makes it possible to maintain and / or increase the biomechanical properties of the hair fiber.

[0211] Example 7: Ex vivo effects of the hydrolyzed extract according to the invention on the biomechanical properties of skin appendages - Antioxidant activity / carbonylation of keratins

[0212] Objective: The aim of this study was to measure the protective effect of the hydrolyzed extract of Silybum marianum seed cake according to the invention against oxidation and in particular carbonylation of keratins and keratin-associated proteins (KAPs: Keratin Associated Proteins).

[0213] Materials and methods:

[0214] Hair strand preparation: Hair braids from Caucasian donors were used for the measurements. The hair was treated with hydrolyzed Silybum marianum seed cake extract according to example la at 0.05% (w / v) in distilled water, by contact for 10 minutes, followed by a washing and drying process. Urban particles (fine particles) were applied to the hair fibers (PM; ERM-CZ100 Fine Dust, Particulate Matter 10-like (PMIO-like), Hydrocarbon Aromatics Polycyclic at 170 qg / cm2), and the hair was immediately exposed to UVA irradiation (84 J / cm2, 6 hours exposure, LED source, emission peak at 365 nm) (stressed hair). Hair treated with water or with 0.2% (w / v) N-acetyl cysteines (antioxidants - positive control) and exposed or not to UVA and particles were used as controls.

[0215] The capillary stems from each experimental condition were sampled, cryopreserved, frozen in liquid nitrogen and maintained at -80°C until analysis.

[0216] a) Quantification of carbonyl proteins by Western blot

[0217] Materials and methods

[0218] Carbonylated proteins were quantified in capillary stems by Western Blot analysis for the different conditions.

[0219] Total proteins (keratins and keratin-associated proteins) were extracted from each hair shaft by a conventional method for those skilled in the art. Total protein quantification was performed using the Bradford method. Carbonylated proteins were labeled using a specific fluorescent probe (Ex=647 nm / Em=650 nm; Baraibar MA, Ladouce R, Friguet B. Proteomic quantification and identification of carbonylated proteins upon oxidative stress and during Cellular aging. J Proteomics. 2013 Oct 30;92:63-70). Proteins were resolved by SDS-PAGE and their fluorescence signal was quantified by acquiring images of the gel using Thermo Fisher Scientific's iBright software system. Total proteins were also stained in the gel for normalization using the iBright system (Thermo Fisher Scientific). A carbonyl score (carbonylated proteins / total proteins), representing the quantification of carbonylated proteins, was obtained for each sample.

[0220] Carbonyl score (sample X) = Fluorescence value of oxidized proteins (sample X) / Fluorescence value of total proteins (sample X)

[0221] A protection index (%) was calculated according to the following formula:

[0222] Protection index % = (Carbonyl score level (stress) - Carbonyl score level (treated with extract according to the invention)) / (Carbonyl score level (stress) - Carbonyl score level (control)) x 100

[0223] For reference, the control group is considered to have maximum effectiveness (100%) and the stress group has minimum effectiveness (0%).

[0224] The threshold for statistical significance was set at 5% (p<0.05).

[0225] Results: The results are summarized in Table 12 below.

[0226] [Tables 12] Carbonyl protein content in hair exposed to oxidative stress (UVA and urban particles) measured by Western blot. Control: Healthy hair. Control: Hair after UVA exposure (84 J / cm² and urban particles). Stressed hair + hydrolyzed extract of Silybum marianum seed cake according to example la at 0.05% (w / v). Stressed hair + N-acetylcysteine ​​at 0.2% (w / v). Mean (%) 76 100 75 89 SD (%) 4 5 1 3 Difference (%) 24 -25 -11 Protection factor > 100% 45% Statistic vs. control (ANOVA) p-value P<0.01 (**) P<0.01 (**) P<0.05 (*)

[0227] A significant increase in protein carbonylation was observed during exposure to stress due to UVA and pollution (UVA 84 J / cm2 and urban particles of type PM 10) (Table 12).

[0228] The hydrolyzed extract of Silybum marianum seed cake according to example 1a at 0.05% (w / v) significantly reduced the level of carbonylated proteins by 25% (p<0.01) in the hair fiber. This corresponds to a protection index >100%.

[0229] b) Quantification of carbonylated proteins by image analysis

[0230] Materials and methods:

[0231] In situ densitometric analysis of the carbonylation signal was performed by analyzing images obtained using Thermo Fisher's "iBright" software and processed with Rasband's ImageJ software. Oxidation levels for each experimental group in the cuticle and cortex regions are shown as mean (UFR) ± standard deviation from the mean (Tables 13 and 14).

[0232] The threshold for statistical significance was set at 5% (p<0.05).

[0233] [Tables 13] Carbonyl protein content in the cuticle of hair exposed to oxidative stress (UVA and urban particles) measured by image analysis. Control: Healthy hair Control: Hair after UVA exposure (84 J / cm² and urban particles) Stressed hair + hydrolyzed extract of Silybum marianum seed cake according to example la at 0.05% (w / v) Stressed hair + N-acetylcysteine ​​s at 0.2% (w / v) Mean (%) 44 100 72 66 SD (%) 7 X 5 1 Difference (%) 56 - -28 -34 Protection index - - 49% 61% Statistic vs. control (t-test) p-value P<0.001 (***•) - P<0.01 (**) P<0.001 (***)

[0234] Results: UVA at 84 J / cm2 and urban particles increased the amount of carbonylated proteins in the cortex and cuticle.

[0235] [Tables 14] Carbonyl protein content in the cortex of hair exposed to oxidative stress (UVA and urban particles) measured by image analysis. Control: Healthy hair. Control: Hair after UVA exposure (84 J / cm² and urban particles). Stressed hair + hydrolyzed extract of Silybum marianum seed cake according to example la at 0.05% (w / v). Stressed hair + N-acetylcysteine ​​s at 0.2% (w / v). Mean (%) 47 100 69 69 SD (%) 7 4 4 4 Difference (%) 53 X -31 -31 Protection index - - 59% 58% Statistic vs control (t-test) p-value P<0.001 (***•) - P<0.001 (***) P<0.001 (***•)

[0236] The hydrolyzed extract of Silybum marianum seed cake according to example 1a at 0.05% (w / v) significantly decreased the carbonyl protein content in the cuticle and cortex by 28% (p<0.01) and 31% (p<0.001), respectively, thus maintaining and / or increasing the biomechanical properties of the hair. This corresponds to a protection index of 49% for the cuticle and 59% for the cortex.

[0237] c) Protection of the structural integrity of the hair

[0238] Objective: The structural state of the keratin proteins in the hair shafts obtained according to the protocol described above is evaluated by optical measurement using an XPolar® microscope (KAMAX). The birefringence properties of a hair fiber depend on the structural integrity of the keratins.

[0239] Hair is composed of fibrous keratin proteins. The crystalline structure of keratin fibers gives hair the ability to change the polarization of light passing through it, a property called birefringence. Birefringence properties depend on hair thickness and the crystalline state of the keratin. A decrease in the birefringence parameter (Kindex) indicates structurally damaged keratins. A Kindex value, which is associated with the birefringence properties of the hair fiber, can be determined and compared between damaged and undamaged hair, and between treated and untreated hair conditions.

[0240] Materials and methods:

[0241] Hair segments obtained according to the protocol described above, 1 cm in length (n = 30 per condition), were transferred to compatible slides for birefringence analysis using XPolar technology. This is an imaging device integrated into a microscope, allowing observation and measurement of the polarimetric properties of the samples studied. Reflected intensity images were first taken to measure the hair thickness. Then, the samples were illuminated with polarized light to measure the Kmax. Assuming that the hair has a cylindrical geometry, it is possible to estimate the birefringence value (Kindex) from the Kmax value and the hair diameters using an abacus.

[0242] A value for improving hair structure was obtained using the damaged group and the untreated groups as references, according to the formula: (stressed hair treated with the hydrolyzed extract according to the invention - stressed hair) / (healthy hair - stressed hair) *100.

[0243] [Tables 15] Birefringence index (Kindex) related to hair structural quality. Control: Healthy hair. Control: Hair after UVA exposure (84 J / cm² and urban particles). Stressed hair + hydrolyzed extract of Silybum marianum seed cake according to example la at 0.05% (w / v). Mean (%) 124 100 114 SD (%) 31 23 37 Difference (%) -24 -14 Improvement index XX 59% Statistic vs. control (t-test with Welch's correction) p-value P<0.001(*) - P<0.05(*)

[0244] Results: Oxidative stress caused a structural change in the hair fiber. A decrease in Kindex values ​​was observed in damaged hair compared to the control of undamaged hair (Table 15). This corresponds to a degradation of the keratin structure by oxidation of keratins under exposure to UVA and urban particles.

[0245] Hair fibers treated with the hydrolyzed extract of Silybum marianum seed cake according to example 1a at 0.05% (w / v) in distilled water, showed a significant increase in the average birefringence value compared to Damaged hair fibers (Table 15). The hydrolyzed extract of Silybum marianum seed cake, as described in example 1a, increased the birefringence index by 14% (p<0.05). This corresponds to a 59% improvement in hair structure compared to damaged hair (improvement index) and therefore indicates an improvement and / or maintenance of the biomechanical properties of the hair fiber.

[0246] d) Integrative hair protection score

[0247] The Hair Protection Score (SPC) is the integrative index for classifying the effectiveness of hair care products. Score values ​​are categorized as: SPC 15; SPC 30; SPC 50; and SPC 50+. The SPC value is obtained using a specific algorithm that integrates early molecular events to assess the protection of hair fibers against everyday damage. Molecular damage, assessed by evaluating protein carbonylation, and structural damage, assessed by evaluating hair birefringence according to Example 7, were integrated to obtain the product's SPC, supporting effective protection of the hair fiber against pollution-induced damage (particles and UVA irradiation). An internal reference was used as a control (N-acetylcysteine: SPC = 50).

[0248] The hydrolyzed extract of Silybum marianum seed cake according to Example la showed a hair protection score of 50+. This score is higher than that of N-acetylcysteine.

[0249] Statistics: All data were analyzed using GraphPad Prism 9 (Insightful Science USA) or SigmaPlot (Systat Software Inv. USA) software. Statistical analyses were performed using binary comparisons of Student's t-tests or ANOVA between groups and damaged conditions. The threshold for statistical significance was set at 5% (p<0.05).

[0250] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the invention protects keratins and keratin-associated proteins from carbonylation induced by exposure of hair fibers to UV and pollutants, allowing the maintenance of their structural integrity and the maintenance of their biomechanical properties.

[0251] Example 8: Penetration of the hydrolyzed extract according to the invention into the fiber (cortex) (FT-IR) on hair fibers (ex vivo)

[0252] Objective: Vibrational spectroscopic techniques such as Raman infrared spectroscopy and Fourier transform infrared spectroscopy (FT-IR) are increasingly used in cosmetic science to study the chemical composition of hair and changes in keratin structure under different environments and treatments. These techniques are non-destructive and require no sample labeling. FT-IR is particularly advantageous for analyzing pigmented hair because it is less prone to intense melanin absorption under visible excitation. The aim of this study is to evaluate the distribution of the constituents of the hydrolyzed extract according to the invention within the hair fiber and to investigate its effect on the infrared profile of keratin.

[0253] Materials and methods:

[0254] Hair preparation and processing: The test was performed on 3 strands of persulfate-bleached (ultra-bleached) Caucasian human hair. The hair strands were standardized (2 g / 2 g / 15 cm), cleaned, rinsed, and dried at room temperature prior to testing. Braids of healthy Caucasian hair (natural dark brown hair) were used as a control of natural, undamaged hair. The hair strands were completely coated with the hydrolyzed extract according to example, mixed at 5% w / v in distilled water, or with water as a placebo. Excess product was removed from the strands by wiping each strand between the gloved thumb and forefinger. The hair was cut into 10 µm sections for infrared imaging.

[0255] Infrared Spectroscopy: The product was characterized for its vibrational spectral profile by infrared spectroscopy coupled with a microscope. Images were collected between 800 and 4000 cm¹, with a pixel size of 6.25 x 6.25 pm². Analytical preprocessing was performed to clean the spectra. The product spectrum was tracked in the hair fiber using a fitting method.

[0256] Results: The hydrolyzed extract according to example la exhibits specific spectral bands at 993, 1053, 1110, 1403, and 1597 cm⁻¹ that are distinct from the keratin spectrum. These spectral markers were used to identify the hydrolyzed extract in the hair fiber. We observe that the hydrolyzed extract is concentrated mainly at the hair cuticle and diffuses into the fiber cortex.

[0257] Example 9: Effects of the hydrolyzed extract according to the invention on the bio-mechanical properties of skin appendages - Molecular restructuring and stabilization of cysteine ​​groups (FT-IR) on hair fibers (ex vivo)

[0258] Objective: The objective is to study more precisely the vibrational spectra of damaged and undamaged keratins, treated and untreated with the hydrolyzed extract of Silybum marianum seed cake according to example 1a. These analyses have demonstrated the interaction of the hydrolyzed extract according to the invention with damaged keratins of the hair cortex, and in particular its general stabilizing effect, and in particular that of cysteic acid-type groups and their derivatives.

[0259] Materials and methods

[0260] Strands of Caucasian brown hair have been oxidized by The wicks were bleached by being calibrated (12 cm / 1 g), cleaned and dried for the study. The wicks were bleached with a 5% v / v hydrogen peroxide solution for 20 minutes, then rinsed and dried.

[0261] The comparative analysis of the vibrational spectra of keratins from undamaged hair and bleached hair treated with or without the extract according to Example (mixed at 5% w / v in distilled water) was performed by "fitting" analysis: a method using the Matlab environment to program the comparison of given infrared spectra of the control with the infrared spectra of the product. Changes in the keratin signal were analyzed using hierarchical cluster analysis to determine the degree of similarity between the spectra. The area under the curve was determined for the absorption bands at 1174 cm¹ and 1116 cm¹.

[0262] The threshold for statistical significance was set at 5% (p<0.05) when comparing the values ​​between them.

[0263] Results:

[0264] Oxidative damage caused by chemical bleaching of hair has shown impacts on the chemical and structural composition of hair with observable changes in vibrational spectrum profiles.

[0265] Hierarchical cluster analysis showed that damaged hair treated with the hydrolyzed extract of Silybum marianum seed cake according to example 1a and virgin hair exhibit spectral profiles that are closer than similar to those of damaged hair. These results suggest that the hydrolyzed extract of Silybum marianum seed cake according to the invention interacts with the damaged keratins of the fiber cortex and helps to stabilize them.

[0266] The spectral bands relating to the integrity of cysteine ​​bonds were then studied in more detail. These were modified under conditions damaged by chemical oxidation compared to healthy, undamaged hair.

[0267] In particular, the spectral bands at 840, 1116, 1150 and 1174 cm 1 (referring respectively to NH out of plane bending, SO2 and CO stretching and SO3 asymmetric stretching) increased in intensity in hair bleached oxidized compared to healthy hair.

[0268] It has been demonstrated that the hydrolyzed extract according to the invention was capable of reducing these band intensities to those of virgin hair.

[0269] Finally, it has been demonstrated that the hydrolyzed Silybum marianum seed cake extract according to the invention was able to reduce these band intensities to approach the spectra of healthy hair (Table 16).

[0270] In particular, oxidative damage has affected the SO, SO2, NH and CO (significant increase in the intensity of these bands). The formation of these groups occurred through the oxidation of the groups and the cleavage of the cysteine ​​disulfide bridges. The hydrolyzed extract of Silybum marianum seed cake according to the invention showed a significant decrease in the intensity of these SO, SO2, NH, and CO bonds.

[0271] The protection index is calculated according to the following formula: (Glycated control -Glycated +Treatment with the extract according to the invention) / (non-glycated control -glycated control) xlOO.

[0272] [Tables 16] Absorption Bands Area under the curve for absorption bands at 1174 cm¹, 1116 cm¹ Undamaged hair controls Bleached hair (untreated) Bleached hair treated with hydrolyzed Silybum marianum seed cake extract as per example at a concentration of 5% w / v 1174 cm¹ (asymmetric stretch of cysteic acid (S=O / SO3)) Mean 0.46 0.74 0.70 Standard deviation 0.05 0.04 0.05 Statistics vs Bleached hair <0.001 (***) X <0.05 (*) Difference % X -5.4% vs untreated % efficacy 14% 1116 cm¹ (SO2 stretch) Mean 1.06 1.46 1.07 Standard deviation 0.15 0.28 0.32 Statistics vs. Bleached Oxidized Hair <0.01 (**) X <0.05 (*) % Difference -26.7% vs. untreated % effectiveness 97.5% 1150 cm1 (related to CO2 vibrators) Mean 0.55 0.76 0.62 Standard deviation 0.09 0.10 0.10 Statistics vs. Bleached Oxidized Hair <0.01 (**) X <0.01 (**) % Difference -18.4% vs. untreated % effectiveness 66.7% 840 cm³ (related to NH₃) Mean 0.07 0.15 0.09 Standard deviation 0.02 0.06 0.05 Statistics vs. bleached hair <0.05 (*) X <0.05 (*) Difference % -40% vs. untreated % effectiveness 75%

[0273] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the invention is capable of interacting with and stabilizing the molecular entities of keratins and keratin-associated proteins in hair fibers damaged by chemical oxidation. The hydrolyzed extract of Silybum marianum seed cake according to the invention provides structural restoration efficacy and therefore repair and / or maintenance and / or enhancement of biomechanical properties, including strength, plasticity, resistance, and / or flexibility, ranging from 14% to 98% depending on the chemical groups for strips between 840 and 1174 cm¹.

[0274] Example 10: Ex vivo effects of the hydrolyzed extract according to the invention on the repair of the hair fiber

[0275] The aim of this study is to measure the repairing effect of the hydrolyzed extract of Silybum marianum seed cake according to the invention on hair fibers damaged by hydrogen peroxide by calorimetry (DSC = Differential Scanning Calorimetry)

[0276] Materials and methods:

[0277] Dark brown Caucasian hair strands were calibrated (1 g; 12 cm) and prepared for the study. The hair strands were washed and then bleached three times with a hydrogen peroxide solution (5.6% H₂O₂ by volume + 13.9% (NH₄)₂S₂O₈ by volume, pH 9.4) for 30 minutes. Healthy (unbleached) hair strands were kept as a control. After washing and drying for 45 minutes at 55°C under airflow, the hair strands were soaked for 24 hours in distilled water (as a control) or in an aqueous solution containing 1% (w / v) hydrolyzed extract of Silybum marianum seed cake according to Example 1b. Both solutions were pre-buffered to pH 5.5. The wicks were rinsed and then dried at approximately 60°C under an airflow.

[0278] The denaturation temperature of human proteins was determined by differential scanning calorimetry according to the method described by FJ. Wortmann, H. Deutz, Characterizing keratins using high-pressure differential scanning calorimetry (HPDSC), Vol.48-Issue 1, p.137-150, 5 April 1993 with a heating rate of 2 K / min is a differential enthalpy analyzer (DSC Q100, TA Instruments).

[0279] The repair index versus placebo is calculated as follows: (bleached hair treated with hydrolyzed extract according to the invention - Control + water (placebo)) / (control healthy hair - Control + water (placebo)) xlOO.

[0280] The repair index versus placebo is calculated as follows: (bleached hair treated with hydrolyzed extract according to the invention - Control bleached hair) / (control bleached hair - Control + water (placebo)) xlOO.

[0281] [Tables 17] Hair fiber temperature differential according to treatments Healthy hair (unbleached, untreated) Bleached control hair (damaged untreated control) Bleached control hair + water (placebo) Bleached hair + hydrolyzed Silybum marianum seed cake extract according to example 1b at 1% (w / v) Mean (°C) 140.4 130.8 132.0 137.4 SD (°C) 0.07 0.16 0.10 0.09 Difference (°C) - - 9.6 vs unbleached - 5.4 vs placebo Repair index (%) - - - 65% vs placebo 69% vs untreated Statistics vs bleached control hair (ANOVA) N=6 p-value <0.001 (***) vs unbleached <0.001 (***) vs placebo

[0282] Results:

[0283] Hair bleached with hydrogen peroxide is more unstable compared to healthy, unbleached hair (Table 17). The difference in the denaturation temperature of hair fiber proteins was -9.6°C between damaged and healthy hair.

[0284] Treating damaged hair with Silybum marianum seed cake extract according to Example 1b at 1% (w / v) improved the stability of hair proteins against heat denaturation by +5.4°C compared to damaged hair treated with water. The hydrolyzed extract of Silybum marianum seed cake according to example 1b showed a repairing effect on hair fiber damaged by chemical oxidative stress compared to healthy hair of 64%.

[0285] Conclusion: Treatment of peroxide-damaged hair with the hydrolyzed extract of Silybum marianum seed cake according to the invention improved the heat resistance of hair fiber proteins. The hydrolyzed extract of Silybum marianum seed cake according to the invention demonstrated a repairing effect on hair fibers damaged by hydrogen peroxide (oxidative stress), thus showing efficacy in repairing and / or maintaining and / or enhancing biomechanical properties, particularly the strength, plasticity, resistance, and / or elasticity of the hair fiber.

[0286] The cortex repair efficacy of the hydrolyzed extract of Silybum marianum seed cake according to the invention at 1% (w / v) is 65% compared to the damaged control treated with water (placebo) and 69% compared to the untreated damaged control.

[0287] Example 11: Ex vivo effects of the extract according to the invention on the strengthening of the hair fiber (single fiber fatigue test)

[0288] Objective: The effect of the hydrolyzed extract of Silybum marianum seed cake, according to Example 1a, on strengthening weakened hair fibers was evaluated by fatigue tests on isolated hair fibers. Fatigue experiments assess the tendency of hair to break under the repeated application of force (Evans TA., Fatigue testing of hair—a statistical approach to hair breakage. J Cosmet Sci. 2009 Nov-Dec;60(6):599-616). The mechanical tensile properties of hair reflect the internal state of its structure. This approach can be considered a more realistic simulation of consumer practices where grooming represents such a stimulus. This method also allows for the evaluation of the repairing effect of a cosmetic ingredient, particularly on the cuticle.

[0289] Materials and methods:

[0290] Strands of dark brown Caucasian hair, bleached three times with hydrogen peroxide, were calibrated (lg / 12 cm) and cleaned for the study. The strands were treated or untreated with 1% (w / v) of Silybum marianum seed cake extract, as described in example la, mixed with distilled water, for 1 hour at pH 5.0–5.5, then rinsed and dried. Between 50 and 60 fibers were studied under standardized conditions of 40% ± 5% relative humidity at 21°C ± 2°C for each treated or untreated condition.

[0291] In this test, individual hair fibers were subjected to repeated deformations. The instrument counts the number of repetitive cycles of a predefined stimulus which are necessary to induce rupture. A maximum number of cycles has been defined. Under these test conditions, the fiber may or may not break during the experiment. A survival probability at a given time was calculated using the Kaplan-Meier survival curve principle to estimate the survival (or keratin fiber that does not break).

[0292] Statistical analyses comparing the groups were performed using a non-parametric model. Differences were considered significant for a p-value < 0.05.

[0293] A hair strengthening index was calculated according to the following equation: (number of cycles before breakage of treated bleached hair - number of cycles before breakage of untreated bleached hair) / (number of cycles before breakage of healthy hair - number of cycles before breakage of untreated bleached hair) x 100

[0294] [Tables 18] Number of cycles before hair breakage Healthy hair (unbleached) Bleached hair control (damaged control) Bleached hair + hydrolyzed extract of Silybum marianum seed cake as per example at 1% (w / v) Average 3380 495 1437 ET 281 74 211 Hair fiber strengthening index - - 33% Statistic vs bleached hair control N=59-73 p-value <0.001 (***•) - <0.001 (***)

[0295] Results:

[0296] Chemical oxidation by hair bleaching significantly reduced the number of cycles inducing fiber breakage (Table 18). This suggests that bleached hair fiber is more fragile and more prone to breakage.

[0297] Treating bleached hair 3 times with hydrolyzed Silybum marianum seed cake extract according to example la at 1% (w / v) significantly improved the number of cycles before breakage, indicating strengthening and / or repair of hair damaged by chemical hair oxidation.

[0298] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the invention exhibits fiber strengthening and therefore repair properties capillary allowing to reduce its probability of breaking and therefore to maintain and / or increase the strength and / or plasticity and / or resistance and / or flexibility of the hair fiber, in particular to repair skin appendages preferentially hair.

[0299] Example 12: Ex vivo effects of the hydrochloric acid extract according to the invention on the biomechanical properties of the hair fiber - Preservation of color (colorimetry)

[0300] Objective: The antioxidant and color-protecting effect of the hydrolyzed extract of Silybum marianum seed cake according to the invention was evaluated in a hair color fading test after sun exposure.

[0301] Materials and methods:

[0302] Strands of Caucasian hair, bleached twice with hydrogen peroxide and colored with a red pigment, were used. The hair strands were calibrated (1 g / 15 cm), cleaned, and dried for the study. The strands were treated or not with 1% (w / v) of Silybum marianum seed cake extract, as described in example, mixed with distilled water, for 1 hour and then dried overnight at room temperature. The strands were exposed to sunlight (Suntest CPS+) at wavelengths between 300 and 800 nm with a lamp power of 592 W / m² for 19 hours. The total exposure energy was 41,469 kJ / m² per cycle. After exposure, the strands were washed for 3 minutes and dried overnight in a chamber conditioned at 40% relative humidity and a temperature of 21°C, followed by colorimetric measurements according to the L*a*b* method.The parameters measured were delta E* (overall color change) and delta a* associated with red / green hues (change in red color). The color of the strands was measured after 1, 2, or 3 cycles of treatment and sun exposure. Eight strands were used per condition.

[0303] Statistical analyses of the data were performed by comparing the groups using Student's t-test. Differences were considered significant for a p-value < 0.05.

[0304] A percentage of hair color preservation efficacy against discoloration by sunlight was obtained using the colored stressed group and untreated groups as references, according to the formula: 100 - [(colored hair treated with the hydrolyzed extract according to the invention / colored control hair)* 100]

[0305] [Tables 19] Delta a* values ​​(red color change) Control colored hair exposed to sunlight Untreated Colored hair exposed to sunlight + hydrolyzed Silybum marianum seed cake extract as per example la at 1% (w / v) 1 treatment and sun exposure cycle Average -6.6% -0.8% AND 1.7% 1.6% Statistical vs Untreated colored hair N=8 p-value <0.05 (*) Difference -5.8% % efficacy -87.9% 2 treatment and sun exposure cycles Average -15.5% -11.4% AND 1.3% 1.4% Statistical vs Untreated colored hair N=8 p-value <0.05 (*) Difference -4.1% % efficacy -26.5% 3 treatment and sun exposure cycles Average -21.1% -14.6% AND 1.3% 1.7% Statistical vs Untreated colored hair N=8 p-value <0.05 (*) Difference -6.5% % efficiency - 30.8%

[0306] Results:

[0307] Colored hair exposed to solar irradiation showed increasing variations in overall color and red color during exposure cycles (Tables 19 and 20). Solar exposure bleached the hair.

[0308] The treatment of coloured hair with the hydrolyzed extract of Silybum marianum seed cake according to example la made it possible to significantly slow down these colour changes (Delta a* and Delta E*) depending on the exposure cycles.

[0309] The hydrolyzed extract of Silybum marianum seed cake according to the invention has shown a protective effect on hair color against discoloration by sunlight and therefore makes it possible to maintain and / or increase the color and / or shine of the hair fiber.

[0310] Example 13: Ex-vivo effects of a hydrolyzed extract according to the invention on the biomechanical properties of hair - anti-breakage

[0311] Objective: The effectiveness of a hair treatment in preventing breakage and split ends was tested on a device allowing repeated combing of hair strands. The influence of chemical hair treatments such as bleaching on hair breakage was examined. Then, the protective effects of reference products on the market and of the hydrolyzed extract according to the invention were studied.

[0312] Materials and methods:

[0313] Strands of Caucasian brown hair were calibrated (12 cm / 1 g), cleaned, and dried for the study. The strands were bleached with a 5% v / v hydrogen peroxide solution for 20 minutes, then rinsed and dried. The strands were treated with the shampoo having the composition below, containing the hydrolyzed extract as a cosmetic ingredient according to example (representing 0.25 g of shampoo / g of hair), and then treated with the conditioner having the composition below, containing the hydrolyzed extract as a cosmetic ingredient according to example (representing 0.125 g of conditioner / g of hair). The products were rinsed out, and the strands were left to rest for 5 hours at 40% relative humidity before analysis. The strands were styled 50,000 times under the following conditions: 40% relative humidity at 30°C. Broken fibers less than 9 cm in length were collected and weighed..

[0314] The hair breakage value is calculated according to the formula: broken hairs (g) / strands (g) x 100

[0315] Outliers were detected by the Grubbs test with a significance level of 0.05 and removed from the data records.

[0316] Anti-breakage shampoo formula:

[0317] [Tables20] Ingredients % by weight Water 74.15 Sodium Benzoate 0.50 Xanthan Gum 0.45 Decyl Glucoside 12.00 Dicaprylyl Ether, Decyl Glucoside, Glyceryl Oleate 1.50 Sodium Cocoyl Glutamate 8.00 Guar Hydroxypropyltrimonium Chloride 0.20 Citric acid, water 0.10 Hydrolyzed extract as a cosmetic ingredient, for example 0.20 Citric acid and water 2.90

[0318] Anti-breakage conditioner formula:

[0319] [Tables21] INCI Name % by weight Water 86.00 Cetearyl Alcohol, Lecithin, Sodium Cetearyl Sulfate, Glus Oil 9.00 Coco-Caprylate 2.00 Olus Oil 2.00 Sodium Benzoate 0.50 Hydrolyzed extract as a cosmetic ingredient, for example 0.20 Citric acid and water 0.30

[0320] [Tables22] Percentage of broken hairs after repeated styling (50,000 times) Weakened (bleached) hair control (% by weight) Weakened hair + treated with shampoo and conditioner containing the hydrolyzed extract as a cosmetic ingredient according to example at 0.2% (w / w) (% by weight) Average 20.1% 4.5% AND 2.5% 1.7% Reduction in hair breakage vs untreated -78% Statistic vs Untreated bleached hair N=10 p-value <0.05 (*)

[0321] Results: The breakage rate of untreated, weakened hair was estimated at 20.1%, compared to 4.5% for weakened hair treated with shampoos and conditioners containing hydrolyzed Silybum marianum seed cake extract, as shown in example (Table 22). The breakage protection provided by the finished products containing hydrolyzed Silybum marianum seed cake extract, as shown in example, was 78%.

[0322] Conclusion: Good conditioning and breakage-protecting properties were found with the shampoo and conditioner containing the hydrolyzed extract of Silybum marianum seed cake according to the invention.

[0323] Example 14: In vitro effects of a hydrolyzed extract according to the invention on the biomechanical properties - protection and stimulation of metabolism on the papilla of the hair follicle

[0324] Papillary fibroblasts play an important role in orchestrating regulatory signals involved in hair follicle cell metabolism (Reynolds AJ, Jahoda CA, Cultured dermal papilla cells induce follicle formation and hair growth by transdifferentiation of an adult epidermis. Development 115: 587-593; (1992); Matsuzaki T, Yoshizato K, Role of hair papilla cells on induction and regeneration processes of hair follicles. Wound Repair Regen. Nov-Dec;6(6):524-30. (1998)). With age, or in response to environmental stresses, the physiology and metabolism of papillary fibroblasts can be affected, and consequently impact the quality of the resulting keratin fiber.

[0325] Objective: The objective of this study was to evaluate the anti-aging and metabolism-stimulating effect of the hydrolyzed extract of Silybum marianum seed cake, according to example 1a, on papillary fibroblast aggregate culture models derived from hair follicles by measuring ATP. This 3D cell model allows for the preservation of the characteristics of the papilla in vivo, in particular its ability to induce the formation and growth of quality hair (Yang CC and Cotsarelis G, Review of hair follicle dermal cells. J Dermatol Sci. 57(1): 2 (2010)).

[0326] Materials and methods:

[0327] A suspension composed of human hair follicle papilla fibroblasts (40 x 10³ cells) in DMEM with 0.2% v / v growth factors (Mesenchymal Stem Cell Growth Supplement) and the presence or absence of hydrolyzed Silybum marianum extract as described in example 1a at a concentration of 0.01% w / v in distilled water, was placed in a 96-well microplate and centrifuged for 5 minutes at 200 g to form aggregates. The aggregates were incubated at 37°C in a controlled atmosphere (5% CO₂ and 95% relative humidity).

[0328] For the analysis of the parameters of interest, the aggregates are rinsed in a buffered saline solution (PB S) and the cells are separated by incubation in a mixture comprising collagenase A 100mg, and a mixture of 20mL of Trypsin (2.5% v / v) and EDTA (0.02% v / v) with a ratio of 1:1.

[0329] Suspended cells were used to measure ATP by bioluminescence according to the supplier's instructions (Bioluminescence Assay Kit CLS II Roche 11699695001, Sigma Aldrich).

[0330] Results: Treatment of aggregates with hydrolyzed extract of Silybum marianum seed cake according to example 1a at 0.01% by weight / volume increased the total amount of ATP produced (+32% compared to the control; Table 23).

[0331] [Tables23] Effect of hydrolyzed Silybum marianum seed cake extract on ATP production by papillary fibroblasts cultured in aggregates. Papillary fibroblast aggregates in a control medium. Papillary fibroblast aggregates treated with hydrolyzed Silybum marianum seed cake extract according to example 1a at 0.01% w / v. Mean (%) 100 132 SD (%) 19 23 Differences (%) - 32 Statistic vs control t-test N=7-10 p-value - <0.01 (**) vs control

[0332] Conclusion: Treatment of papilla fibroblast aggregates with the hydrolyzed extract of Silybum marianum seed cake according to the invention increases ATP within the pseudo-papillum. These results suggest a vitalizing potential for the hair follicle, leading to structural and functional strengthening of the keratin fiber.

[0333] Example 15: Consumer test with shampoo and conditioner comprising respectively the hydrolyzed extract as a cosmetic ingredient according to example the

[0334] Objective: The aim of the study is to evaluate consumer perception of the benefits provided by the hydrolyzed extract as a cosmetic ingredient, incorporated into a shampoo and conditioner for damaged hair.

[0335] Materials and methods:

[0336] The study was randomized double-blind with a total of 31 volunteers. The shampoo and conditioner used were identical to those used in Example 13 (Tables 20 and 21).

[0337] The same volunteers applied either the shampoo and conditioner containing the hydrolyzed extract as a cosmetic ingredient or the placebo products at least 3 times a week for a period of 4 weeks, sequentially, with a 3-week treatment break between the two treatment phases.

[0338] The volunteers completed a self-perception questionnaire on consumer benefits measuring their opinions on the products, after 4 weeks of use. The number of positive or negative questions were grouped and analyzed using a non-parametric binomial test (Chambers E. and Wolf M. “Sensory Testing Methods: Second Edition.” Micro & Nano Letters (1996)). The significance level was set at 5% (p<0.05).

[0339] Results:

[0340] A significant majority of volunteers perceived beneficial effects on their hair from regularly using a shampoo and conditioner containing the hydrolyzed extract as a cosmetic ingredient. Among these positive opinions, using this hygiene and care routine was perceived as providing protection against chemical stress, breakage, and split ends. Hair appeared less damaged and healthier-looking. Hair seemed to show more vitality, volume, shine, and less dryness. The scalp felt more comfortable.

[0341] On the other hand, the placebo shampoo and conditioner, not containing the hydrolyzed extract as a cosmetic ingredient, did not show a significant majority of favorable opinions regarding the perception of a benefit on the protection of hair against chemical stress, against split ends, the increase in vitality, volume and the reduction of hair dryness.

[0342] [Tables24] Percentage of favorable opinions on the consumer benefits of using rinse-off hygiene and hair care products. Placebo shampoo and conditioner Shampoo and conditioner containing the hydrolyzed extract as a cosmetic ingredient, according to Example, at 0.2% (W / W) Using the shampoo and conditioner protects my hair against... chemical stress 52% 71%* ...breakage 69%* 74%* ...split ends 62% 77%* My hair seems... less damaged 69%* 77%* ...healthier looking 72%* 81%* Using the shampoo and conditioner improves... the vitality of my hair 62% 74%* ...volume 59% 71%* ...dryness 62% 71%* ...shine 72%* 81%*

[0343] * Significant majority of favorable opinion (p<0.05) according to the binomial test.

[0344] Conclusion: Additional benefits of the presence of the hydrolyzed extract as a cosmetic ingredient on the hair were perceived by volunteers who used a shampoo and conditioner regularly for 4 weeks. Example 16: Cosmetic ingredients

[0345] Cosmetic ingredient 1 f % by weight): Hydrolyzed extract (as in example a) 20% Maltodextrin 80%

[0346] Cosmetic ingredient 2 (% by weight): Hydrolyzed extract according to example 1b) 30% Maltodextrin 70%

[0347] Example 17: Cosmetic compositions comprising the cosmetic ingredient

[0348] Anti-breakage shampoo formula

[0349] [Tables25] Ingredients % Water 74.15 Sodium Benzoate 0.50 Xanthan Gum 0.45 Decyl Glucoside 12.00 Dicaprylyl Ether, Decyl Glucoside, Glyceryl Oleate 1.50 Sodium Cocoyl Glutamate 8.00 Guar Hydroxypropyltrimonium Chloride 0.20 Citric acid, water 0.10 Hydrolyzed extract as a cosmetic ingredient, for example 0.20 Citric acid and water 2.90

[0350] Anti-breakage conditioner formula

[0351] [Tables26] Ingredients % Water 86.00 Cetearyl Alcohol, Lecithin, Sodium Cetearyl Sulfate, Glus Oil 9.00 Coco-Caprylate 2.00 Olus Oil 2.00 Sodium Benzoate 0.50 Hydrolyzed extract as a cosmetic ingredient, e.g. 0.20 Citric acid and water 0.30

Claims

Demands

1. Non-therapeutic cosmetic use of a hydrolyzed extract of Silybum marianum seed cake to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

2. Use according to claim 1, characterized in that the hydrolyzed extract is obtained in water as the sole solvent.

3. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract is obtained by enzymatic hydrolysis.

4. Use according to claim 3, characterized in that the hydrolyzed extract is obtained by enzymatic hydrolysis at a pH between 3 and 10, advantageously between 7 and 9.

5. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract has a total protein dry mass content of between 50% and 95% by weight, preferably between 60% and 90% by weight, more preferably between 60% and 70% by weight, relative to the total dry mass weight of the hydrolyzed extract.

6. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract has a dry mass content of peptides with an average molecular weight by weight of 10kDa to 20kDa of between 50% and 90%, advantageously between 65% and 90%, more advantageously between 75% and 90%, by weight relative to the total dry mass weight of peptides in the hydrolyzed extract.

7. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract has a dry mass content of peptides with a weight average molecular weight between 1OODa and 700 Da of between 60% and 90%, advantageously between 70% and 80%, more advantageously between 75% and 80%, by weight relative to the total dry mass weight of peptides with a weight average molecular weight between 1OODa and 7000 Da of the hydrolyzed extract.

8. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract maintains and / or increases the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e., the non-brittle appearance, and / or suppleness of skin appendages, preferably keratin fibers, advantageously hair pain.

9. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract is intended for topical application to all or part of the skin of the body and / or face including skin appendages, in particular the scalp, and / or to all or part of the skin appendages, advantageously on the nails, hair, body hair, in particular beard hair, eyelashes and / or eyebrows, more advantageously on all or part of the keratin fibers, preferably on all or part of the hair.

10. Use according to claim 9 wherein the application is made on damaged, colored or highlighted, dull, dry, brittle, fragile, friable, fine, split and / or stressed hair.

11. Use according to any one of the preceding claims, characterized in that the hydrolyzed extract is atomized in the presence of a weight concentration of maltodextrin of between 1% and 99%, advantageously between 5% and 90%, preferably between 40% and 80%, again preferably between 70% and 80%, relative to the total weight of the powder obtained.

12. Use according to any one of the preceding claims characterized in that the hydrolyzed extract is present in a cosmetic composition, at a content of between 1x104% and 10% by weight, preferably from 1x103% to 5% by weight, more preferably from 1x103% to 3% by weight, even more preferably from 1x103% to 1% by weight, advantageously from 0.01% to 1% by weight, relative to the total weight of the cosmetic composition.

13. Use according to claim 12, characterized in that the cosmetic composition comprises at least one cosmetically acceptable excipient and in that this composition is selected from a serum, lotion, cream, shampoo, conditioner, oil, milk, ointment, paste, mousse, emulsion, hydrogel, shower gel, mask, lacquer, spray, wax, mascara, makeup pencil, varnish, advantageously a shampoo, conditioner or lotion.

14. Use according to claim 1, for repairing skin appendages, preferably hair.

15. A non-therapeutic cosmetic treatment method comprising the topical application of a hydrolyzed extract of Silybum marianum seed cake or a cosmetic composition comprising it to maintain and / or increase the biomechanical properties of skin appendages, advantageously keratin fibers, preferably hair.

16. A cosmetic treatment method according to claim 14, for repairing skin appendages, advantageously keratin fibers, preferably hair.

17. A cosmetic treatment method according to claim 15 or 16, characterized in that the hydrolyzed extract or the cosmetic composition comprising it is applied topically to all or part of the skin of the body and / or face including skin appendages, in particular the scalp, and / or to all or part of the skin appendages, preferably on all or part of the keratin fibers, more preferably on all or part of the hair.

18. A cosmetic care process according to any one of claims 15 to 17, characterized in that the extract is as defined in any one of claims 2 to 8 and 11 to 14.