Cosmetic application of hydrolyzed extract of silybum marianum seed cake

Abstract

The present invention relates to the cosmetic use of a hydrolyzed extract of a cake of silybum marianum seeds for maintaining and / or increasing the biomechanical properties of the skin covering. Another subject of the invention consists of a cosmetic care method comprising the topical application of a hydrolyzed cake extract or a composition comprising the same.

Description

Technical Field

[0001] This invention relates to the cosmetic use of hydrolyzed extracts of milk thistle seed cake. Background Technology

[0002] The care of skin coverings (such as nails, body hair, head hair, eyelashes, and eyebrows) involves comprehensive treatment, which includes direct treatment of keratin fibers as well as direct treatment of the corresponding skin areas (including the scalp and hair follicles (or hair bulbs and hair fibers)).

[0003] Furthermore, direct care of hair fibers involves the treatment of their constituent proteins. Hair proteins (such as keratin) are particularly sensitive to damage caused by lifestyle, exposure to chemical hygiene products (such as their nature and frequency of use), styling habits, and environmental conditions (such as sunlight, pollution, salt, and wind), changes in climate, general lifestyle, and age. These proteins denature and make the hair covering less resistant, less soft, more brittle, and more dull, especially causing split ends. These coverings then lose their biomechanical properties.

[0004] One solution is to provide exogenous hydrolyzed proteins to compensate for damage to keratin proteins and improve the moisture retention and elasticity of keratin fibers, especially hair fibers. As a result, these proteins impart greater softness, shine, elasticity, and density to the fibers.

[0005] The hydrolysis of these proteins (chemical, thermal, or enzymatic) is a necessary preliminary method for fragmenting them into low molecular weight fragments, thereby promoting their solubility in water and their penetration into keratin fibers, and facilitating their formulation. Specifically, only molecules with low weight-average molecular weight (less than 7 kDa) can penetrate into keratin fibers, especially damaged hair fibers. They strengthen keratin fibers from the inside, making them stronger, more resilient, less brittle, and especially reducing split ends in the hair fiber.

[0006] High-performance hydrolyzed proteins possess the property of adhering to hair fibers (with weak binding to hair keratin) due to their ionic charge and polar sites (van der Waals interactions). They can also form a protective film on the surface of hair fibers.

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

[0008] Completely unexpectedly, the applicant has discovered that a hydrolyzed extract of milk thistle seed cake possesses the ability to penetrate keratin fibers and thereby increase their biomechanical properties. This hydrolyzed extract has been shown to improve the quality of skin coverings, increasing their resistance and reducing breakage, particularly in hair, making them shinier, shinier, and with greater volume. Furthermore, the hydrolyzed extract of milk thistle seed cake exhibits skin covering repair properties, as will be demonstrated in the following description.

[0009] One advantage of the hydrolyzed extract according to the invention is that it is derived from a byproduct of industrial extraction residues, which are generally not upgraded and are considered industrial waste, thus representing an ecologically responsible approach. Another advantage is that it is an ingredient that provides comprehensive care because it affects both keratin fibers (particularly hair) and hair follicles. The hydrolyzed extract according to the invention can also be readily produced on an industrial scale. Finally, enzymatic hydrolysis enables the fractionation of proteins in the cake to recover low molecular weight peptides, which are the very peptides that can penetrate keratin fibers to provide the cosmetic effects described herein.

[0010] The hydrolyzed extract according to the present invention is derived from the cake of the plant milk thistle. This plant, also known as milk thistle, has been known for thousands of years for its effects on the liver. In Asia and Africa, it is still used to treat kidney problems, gastrointestinal disorders, heart disease, rheumatism, and fever. The seeds are primarily used for these therapeutic purposes.

[0011] In several countries in the Middle East, North Africa, Sardinia, and Spain, the tender stems, seeds, and buds are still routinely consumed. The fruit of the milk thistle can be used for oil extraction or as a coarse flour, mixed with wheat flour, for the preparation of various baked goods or as a food product.

[0012] Milk thistle is also commonly used in cosmetics, especially the flowers (without seeds) to improve skin elasticity, firmness, barrier function, and hydration, and also to reduce hair loss.

[0013] Patent application DE 4323614 discloses an extract of milk thistle fruit used as a medicament for hair growth and regeneration.

[0014] Patent application CN 103446021 discloses an extract of milk thistle seeds obtained by heating and propylene glycol extraction of the whole seeds, which protects hair from damage caused by UV rays, especially through antioxidant effects that protect hair color, while also improving firmness and tensile strength.

[0015] Patent FR3053253 describes a method for preparing an extract of milk thistle achenes containing less than 0.2% silymarin. The method involves extracting the oil using a solvent selected from aqueous solutions, subcritical water, ethanol, isopropanol, or mixtures thereof.

[0016] Therefore, to the best of the applicant's knowledge, no prior art discloses cosmetic uses of the hydrolyzed extract according to the present invention. Nor are such uses suggested, alone or in combination, in any document. Summary of the Invention

[0017] Therefore, the first subject concerns the non-therapeutic cosmetic use of hydrolyzed extracts of milk thistle seed cake for maintaining and / or increasing the biomechanical properties of skin coverage, keratin fibers, and preferably hair.

[0018] The second topic relates to the non-therapeutic cosmetic use of the hydrolyzed extract according to the invention in cosmetic compositions.

[0019] The third topic relates to a non-therapeutic cosmetic care method comprising the topical application of a hydrolyzed extract according to the invention or a cosmetic composition containing the same to maintain and / or increase skin coverage, beneficial keratin fibers, and preferably the biomechanical properties of hair.

[0020] Therefore, the first subject concerns the non-therapeutic cosmetic use of hydrolyzed extracts of milk thistle seed cake for maintaining and / or increasing the biomechanical properties of skin coverage, keratin fibers, and preferably hair.

[0021] The term "hydrolyzed extract" refers to the product obtained by enzymatic or chemical hydrolysis of milk thistle seed cake in an aqueous solvent. In a preferred embodiment, the hydrolyzed extract is obtained by enzymatic hydrolysis.

[0022] The term "cake" refers to a byproduct of seed extraction, i.e., the residue obtained after oil extraction. Preferably, the milk thistle seed cake used is defatted milk thistle seed cake.

[0023] The term "defatted" refers to a cake having an oil content that is strictly less than 10% by weight, and preferably strictly less than 8% by weight, relative to the total weight of the cake.

[0024] Generally, the term "oil" refers to all the lipophilic compounds contained in milk thistle seeds, such as fat-soluble vitamins, fatty acids, monoglycerides, diglycerides, triglycerides, and phospholipids.

[0025] The term "seed" refers to a fruit or achene that does not contain its pericarp. Therefore, the pericarp (also called the pericarp) is not included in the seed according to the invention.

[0026] The term "cosmetic use" means non-therapeutic, non-pharmaceutical, and non-dermatological use, that is, use that does not require or involve therapeutic treatment and is intended for use on healthy skin coverings and / or healthy skin (including skin coverings).

[0027] The term “healthy” refers to skin coverings and / or skin that are classified as nonpathological by experts in the field, dermatologists, meaning the absence of candidiasis, disease or condition, infection, inflammation, scarring, skin diseases or conditions such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, especially seborrheic dermatitis, dandruff, or wounds or injuries and / or other skin diseases and / or hair loss and / or baldness and / or alopecia areata.

[0028] Therefore, for the purposes of this invention, "damaged" skin coverings are those classified as "non-pathological" coverings by experts in the art. Thus, they are healthy while damaged. Specifically, damaged skin coverings are those that have lost their flexibility and / or plasticity and / or resistance and / or strength and / or luster and / or color and / or volume, and are therefore particularly dehydrated. Thus, "damaged" skin coverings lose their biomechanical properties. Therefore, they become dull, dry, less resistant, brittle, fragile, breakable, split, and / or bendable and / or forked. "Damaged" skin coverings may be caused by exposure to stresses such as environmental conditions, such as wind, cold, salt, chlorine, pollution, sunlight, lifestyle, or even age, and also by exposure to chemical or mechanical stresses caused by routine handling of the coverings (such as friction, styling, curling, dyeing, bleaching, straightening, hair dryers, cosmetics or hygiene chemicals such as nail polish, cosmetic and / or makeup removers, shampoos, conditioners, or creams).

[0029] Keratin fibers, especially in hair, become more difficult to style and shape. This loss of hair surface quality is visible and unsightly. Hair also reflects less light and is therefore noticeably less shiny, less pigmented, and less lustrous. Hair is also finer and less dense. This is especially true for dyed or highlighted hair.

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

[0031] The term "keratin fibers" refers to all the fibers that make up the human hair system, especially hair, eyelashes, eyebrows, body hair, and especially beard (including mustache). Preferably, it refers to hair.

[0032] The hydrolyzed extract according to the present invention is a locally acceptable component.

[0033] The term "locally acceptable" means that it is suitable for topical application, is non-toxic and non-irritating to the skin, especially the scalp or skin coverings, does not cause allergic or inflammatory reactions, and is not a chemically unstable ingredient.

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

[0035] The term "topical" means the direct, localized application and / or vaporization of an ingredient on the surface of the skin, particularly the scalp or skin coverings, especially the hair.

[0036] Hydrolyzed extracts can be applied topically to all or some of the surface of the skin (including skin coverings, especially the scalp), or to all or some of the skin coverings, beneficial to nails, hair, body hair, especially beard, eyelashes and / or eyebrows, more beneficial to keratin fibers, and even more beneficial to hair.

[0037] Therefore, one subject of the present invention relates to the hydrolyzed extract of milk thistle seed cake for the maintenance and / or increase of skin coverage, beneficial keratin fibers, more favorable biomechanical properties of hair, and / or for the maintenance and / or increase of their biomechanical properties in a non-therapeutic cosmetic purpose.

[0038] Within the meaning of this invention, the term "repairing biomechanical properties" refers to at least partially improving and / or increasing the biomechanical properties of skin coverings (preferably keratin fibers, advantageously hair) damaged due to exposure to environmental, chemical, biological, and / or mechanical stresses. In particular, repaired skin coverings can restore the biomechanical properties they possessed prior to the damage, i.e., the biomechanical properties of undamaged skin coverings. Therefore, the effect of the hydrolyzed extract according to the invention on these biomechanical properties can be compared, for example, by comparing the biomechanical properties of skin coverings before and after treatment with the hydrolyzed extract according to the invention, or by comparing the biomechanical properties of damaged skin coverings after treatment with the hydrolyzed extract according to the invention with those of undamaged skin coverings.

[0039] For the purposes of this invention, the term "maintaining biomechanical properties" means preventing and / or avoiding the deterioration of the biomechanical properties of skin coverings, preferably keratin fibers, and advantageously hair, particularly when exposed to environmental, chemical, biological, and / or mechanical stresses, especially by preventing skin coverings from becoming damaged and / or by preventing such skin coverings from becoming even more damaged. Therefore, the effect of the hydrolyzed extract according to the invention on these biomechanical properties can be compared, for example, by comparing the biomechanical properties of skin coverings treated with, and exposed to, environmental, chemical, biological, and / or mechanical stresses using the hydrolyzed extract according to the invention, with those of skin coverings not treated with and exposed to environmental, chemical, biological, and / or mechanical stresses using the hydrolyzed extract according to the invention.

[0040] For the purposes of this invention, the term "increased biomechanical properties" means improved biomechanical properties of skin coverings, preferably keratin fibers, advantageously hair, especially skin coverings damaged due to exposure to environmental, chemical, biological, and / or mechanical stresses, as well as undamaged skin coverings. Therefore, the effect of the hydrolyzed extract according to the invention on these biomechanical properties can be compared, for example, by comparing the biomechanical properties of skin coverings before and after treatment with the hydrolyzed extract according to the invention.

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

[0042] Advantageously, the hydrolyzed extract of milk thistle seed cake is used for repairing skin coverings, preferably keratin fibers, and advantageously hair.

[0043] Preferably, the hydrolyzed extract of milk thistle seed cake is used to maintain and / or increase skin coverage, preferably keratin fibers, hair resistance and / or strength and / or volume and / or color and / or shine and / or plasticity (i.e., non-brittle aspects) and / or smoothness.

[0044] These properties can be evaluated in vitro via mechanical testing, in which the resistance of skin coverings, preferably keratin fibers, and more preferably hair, to tensile, stretching, bending, friction, and torsional forces is measured. These biomechanical parameters, especially those of hair, can be evaluated in response to tension and can be measured, for example, via tensile testing (Dia Stron). The measured parameters can be elastic modulus (Pa), elongation at break (%), breaking force (gmf), gradient of the plastic strain region (gmf / % elongation), and normalized relative to the diameter of the skin covering. This last parameter (gradient) allows for the measurement of the plasticity of the material under study just before fracture. This technique provides a stress (or force) curve as a function of the characteristic elongation of the material being tested. It is known that after disulfide bonds in the cortical structural domains break into free radicals, an increase in elongation at break and a decrease in the plastic strain gradient can be observed in hair fibers (outer skin layer) damaged by oxidation, reduction, or ultraviolet radiation.

[0045] In one embodiment of the invention, "maintaining and / or increasing the biomechanical properties of skin coverings" means reducing advanced glycation end products (AGEs), which occur particularly during time-induced or light-induced aging and also daily in response to environmental conditions and contact with chemical or physical oxidants. These glycation products reduce the biomechanical properties of skin coverings.

[0046] Environmental stresses include, in particular, smog (especially cigarette smoke), pollution (especially metals, PM2.5 and PM10 fine particles), temperature (especially heat and cold and sudden temperature changes), water content (especially humidity or aridity), 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), and inherent and / or time-induced aging. The latter also contributes to the loss of structural and / or functional quality of skin coverings, preferably keratin fibers, especially eyelashes, eyebrows and hair, and more particularly hair.

[0047] Chemical stress includes chemical agents such as corrosive convenience agents, hygiene and beauty products, especially corrosive shampoos and hair care and / or hair treatment products, particularly those used for styling, setting (such as straightening and / or permanent wavy hair) and / or for dyeing and / or bleaching, chlorine and transition metals present in swimming pool water, nail polish and / or solubilizers, solvents or cosmetic products.

[0048] Mechanical stresses include friction such as brushing and / or rubbing fabrics (pillows or clothes) and / or particulate matter such as dust and / or sand, heating by hair dryers and straighteners, and / or styling, especially exposure to tension, tensile force and / or torsional force.

[0049] Therefore, in one embodiment of the invention, the ability of the extract according to the invention to "repair" damaged skin coverings can be evaluated according to the prior art. Conventional methods for measuring repair effects allow the evaluated product to restore the visual, structural, and / or functional state of the damaged skin covering, preferably damaged hair, to be measured, a state equivalent to that of undamaged skin coverings, preferably undamaged hair. These are performed on damaged (or damaged) skin coverings (preferably hair), and the repair effect is measured by comparison with undamaged (or undamaged) skin coverings (preferably hair). Advantageously, the repair effect is the repair effect on skin coverings damaged (or damaged) in vitro by an oxidant (which induces denaturation of the proteins in the covering). Advantageously, this is the repair effect of the hydrolyzed extract according to the invention on hair, and more preferably, the oxidant is hydrogen peroxide. More advantageously, the protein denaturation was evaluated by measuring the protein denaturation temperature (°C) using differential scanning calorimetry (DSC Q100, TA Instruments) under the conditions described in Example 10, by differential calorimetry (Wortmann and Deutz, Appl. Polym. Sci., 48, 137-150 (1993)).

[0050] To evaluate the repair effect on skin appendages, particularly on hair, in vivo, several methods selected from video microscopy, confocal microscopy, FTIR or Raman microscopy, X-ray microscopy, and electron microscopy can be used. These methods are particularly useful for observing the condition and / or quality of the stratum corneum and the protective capsule of keratin fibers in skin appendages. Microscopic study of the entire hair shaft allows the visualization and quantification of isolated scales as evidence of surface damage. These methods can be used on hair cross-sections to further observe the condition and / or quality of the keratin fiber cortex. Physical surface quantification methods, such as atomic force microscopy or white light interferometry, can be implemented to evaluate the morphology of skin appendages, especially hair, for evaluation of chemical (XPS), charge (flow potential), or energy (reversed-phase gas chromatography).

[0051] To evaluate the restorative effect of the hydrolyzed extract according to the present invention on the internal properties of skin coverings, especially hair, a calorimetric method (DSC) can be used.

[0052] Within the meaning of this invention, the term "maintain and / or increase volume" means maintaining and / or increasing the thickness of the skin covering over its 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, the term "maintaining and / or increasing the biomechanical properties of skin coverings" means maintaining and / or increasing their resistance and / or their gloss and / or their color and / or their volume and / or their strength and / or their plasticity, i.e., their non-brittleness and / or their flexibility. These properties can be measured by the methods described in the following paragraphs and corresponding examples:

[0054] - By measuring free radical activity, especially in vitro free radical activity as described in Example 3;

[0055] - By measuring the chelating activity against transition metals such as iron (Fe), especially the in vitro chelating activity as described in Example 4;

[0056] - By measuring the chelating activity of transition metals such as copper (Cu), especially the in vitro chelating activity as described in Example 5;

[0057] - By measuring the anti-glycation activity against proteins in skin coverings, especially the in vitro anti-glycation activity as described in Example 6;

[0058] - By measuring the carbonylation of keratin and related proteins in skin coverings, especially in vitro carbonylation as described in Examples 7a and b);

[0059] - By measuring the hair protection score (HPS) in vitro, as described in Example 7d;

[0060] - As in Example 9, the stabilization of cysteine ​​groups was measured in vitro;

[0061] - Their fracture can be evaluated, as in Examples 11 and 13, by means of in vitro tensile strength testing (Diastron), fatigue testing, or repeated hair styling testing;

[0062] - For example, in Example 12, color changes of skin coverings were measured ex vivo;

[0063] - As in Example 14, the amount of ATP in fibroblasts of the pilosebaceous follicle was measured in vitro.

[0064] These properties can be evaluated in vitro by measuring free radical activity, for example by a DPPH test, in the presence of the hydrolyzed extract according to the invention. Therefore, in a particularly preferred embodiment, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin covering” when the percentage inhibition of free radical DPPH° 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%, and preferably greater than 90% relative to the percentage inhibition of free radical DPPH° oxidation measured in the absence of the hydrolyzed extract according to the invention. Advantageously, the skin covering is keratin fibers, preferably hair.

[0065] These properties can also be evaluated in vitro by measuring chelating activity against transition metals such as iron (Fe), for example with tryptophan degradation tests and / or dityrosine and / or pentosine and / or fluorescent glycosylation product formation tests, which are measured in the presence of hydrogen peroxide and iron as oxidation catalysts after oxidation of albumin, which is used as a protein model, in the presence of the hydrolyzed extract according to the invention.

[0066] In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when the protection index of the tryptophan degradation test calculated under the conditions of Example 4 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 cover is keratin fibers, preferably hair.

[0067] In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 4, the percentage of dityrosine formation is reduced by at least 10%, advantageously at least 15%, more advantageously at least 20%, and preferably at least 25% relative to the percentage of dityrosine formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the dityrosine formation test calculated under the conditions of Example 4 is greater than 30%, advantageously greater than 35%, more advantageously greater than 60%, and particularly greater than 65%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0068] In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 4, the percentage of pentosaccharide formation is reduced by at least 10%, advantageously at least 25%, relative to the percentage of pentosaccharide formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the pentosaccharide formation test calculated under the conditions of Example 4 is greater than 50%, advantageously greater than 60%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0069] In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 4, the percentage of fluorescent glycosylation product formation is reduced by at least 5%, advantageously at least 10%, more advantageously at least 13% relative to the percentage of fluorescent glycosylation product formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the test for fluorescent glycosylation product formation calculated under the conditions of Example 4 is greater than 10%, advantageously greater than 15%, more advantageously greater than 20%, particularly greater than 25%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0070] The biomechanical properties according to the invention can also be evaluated in vitro by measuring chelating activity against transition metals (such as copper (Cu)), for example by a lipid oxidation test, by evaluating the formation of malondialdehyde (MDA) in the presence of copper (Cu) (an oxidation catalyst) and thiobarbituric acid (TBA) in the presence of the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 5, the percentage of oxidized lipids obtained by measuring MDA is reduced by at least 20%, advantageously at least 40%, more advantageously at least 60%, preferably at least 75%, compared to the percentage of oxidized lipids obtained by measuring MDA in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the fluorescent glycosylation product formation test calculated under the conditions of Example 5 is greater than 60%, advantageously greater than 75%, more advantageously greater than 90%, particularly greater than 95%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0071] These properties can also be evaluated in vitro by measuring the anti-glycation activity of proteins in the skin covering, for example, by a dityrosine and / or pentasaccharide formation test measured after incubation of the covering in a glucose solution in the presence of the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin covering” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 6, the percentage of dityrosine formation is reduced by at least 5%, advantageously at least 10%, more advantageously at least 15% relative to the percentage of dityrosine formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the dityrosine formation test calculated under the conditions of Example 6 is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and particularly greater than 55%. Advantageously, the skin covering is keratin fibers, preferably hair.

[0072] In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 6, the percentage of pentosaccharide formation is reduced by at least 5%, advantageously at least 10%, more advantageously at least 15% relative to the percentage of pentosaccharide formation measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index of the pentosaccharide formation test calculated under the conditions of Example 6 is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, particularly greater than 55%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0073] Biomechanical properties can also be evaluated in vitro by measuring the carbonylation of keratin and its associated proteins, for example by particle (contamination) and UV testing, in the presence of the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin covering” when, in the presence of the hydrolyzed extract according to the invention, particularly in the stratum corneum, and even more particularly in the cortex, preferably under the conditions described in Example 7, the percentage of carbonylated proteins (oxidation products) in the cortex is reduced by at least 5%, advantageously at least 15%, more advantageously at least 20%, preferably at least 25%, more advantageously at least 30%, relative to the percentage of carbonylated proteins measured in the absence of the hydrolyzed extract according to the invention, and / or when the protection index calculated under the conditions of Example 7 is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, particularly greater than 55%. Advantageously, the skin covering is keratin fibers, preferably hair.

[0074] Biomechanical properties can also be evaluated in vitro by measuring the oxidative stress of the skin covering, for example by optically measuring the birefringence properties, particularly the birefringence index (K-index), of the skin covering treated with the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin covering” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 7c, the K-index value increases by at least 5%, advantageously at least 10%, more advantageously at least 13% relative to the K-index value measured in the absence of the hydrolyzed extract according to the invention, and / or when the index calculated under the conditions of Example 7c increases by more than 40%, advantageously more than 45%, more advantageously more than 50%, particularly more than 55%. Advantageously, the skin covering is keratin fiber, preferably hair. More advantageously, it is the hydrolyzed extract prepared according to Example 1a).

[0075] Biomechanical properties can also be evaluated in vitro by measuring the Hair Protection Score (HPS) to classify hair care products applied to damaged skin coverings (UVA irradiation and particulate matter) treated with the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is preferably effective for “maintaining and / or increasing the biomechanical properties of the skin covering” when the HPS is greater than 30, preferably greater than or equal to 50. Advantageously, the skin covering is keratin fibers, preferably hair.

[0076] These properties can be further evaluated in vitro by measuring the stabilization of cysteine ​​groups, for example by measuring the vibrational spectrum of keratin in damaged skin coverings treated with the hydrolyzed extract according to the invention. In alternative embodiments, when in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 9, the 1174, 1116, 1150 and / or 840 cm⁻¹ of the damaged skin coverings are... -1 The absorption bands were 1174, 1116, 1150, and / or 840 cm⁻¹ relative to those measured for bleach-damaged skin coverings in the absence of the hydrolyzed extract according to the invention. -1 The amount of hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of the skin cover” when the absorption band is reduced by at least 5%, advantageously at least 15%, more advantageously at least 20%, or even more advantageously at least 35% and / or when the efficacy index calculated under the conditions of Example 9 is greater than 5%, advantageously greater than 10%, and more advantageously greater than 13%. Advantageously, the skin cover is keratin fibers, preferably hair.

[0077] Biomechanical properties, particularly strength and / or plasticity and / or resistance and / or flexibility, can also be evaluated in vitro by measuring the stability of the damaged skin covering, for example by measuring the denaturation temperature of the proteins in the damaged skin covering using hydrogen peroxide, and then treating it with the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the strength and / or plasticity and / or resistance and / or flexibility of the skin covering” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 10, the denaturation temperature increases by at least 2°C, advantageously at least 5°C, relative to the denaturation temperature measured in the absence of the hydrolyzed extract according to the invention, and / or when, under the conditions of Example 10, the repair index relative to placebo-treated or untreated skin coverings is greater than 40%, advantageously greater than 45%, more advantageously greater than 50%, and particularly greater than 60%. Advantageously, the skin covering is keratin fibers, preferably hair.

[0078] These properties can also be evaluated by tensile strength testing (Diastron), fatigue testing, or repeated hair styling tests that allow for evaluation of breakage. Hair fibers with split ends can also be quantified. Furthermore, these biomechanical properties can be assessed by measuring the styling force required for dry or wet hair. Notably, these properties can be evaluated in vitro by measuring the breakage of the skin covering, for example by fatigue testing of damaged skin coverings, preferably treated with hydrogen peroxide and then with the hydrolyzed extract according to the invention. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the strength and / or plasticity and / or resistance and / or smoothness of the skin covering” when, in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 11, the number of cycles before breakage of the skin covering or the hair fiber strengthening index is greater than 10%, preferably greater than 20%, more preferably greater than 30% compared to the number of cycles before breakage or the strengthening index of the skin covering measured without treatment with the hydrolyzed extract according to the invention. Advantageously, the skin covering is keratin fiber, preferably hair. More advantageously, it is a hydrolyzed extract prepared according to Example 1a).

[0079] In alternative embodiments, when, after repeated shaping of the damaged skin covering and in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 13, the reduction in the percentage of breakage is greater than 50%, preferably greater than 70%, and more preferably greater than 75% relative to the percentage of breakage of the skin covering measured without treatment with the hydrolyzed extract according to the invention, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the strength and / or plasticity and / or resistance and / or flexibility of the skin covering.” Advantageously, the skin covering is keratin fiber, preferably hair. More advantageously, the hydrolyzed extract is prepared as according to Example 1c).

[0080] The biomechanical properties of color and / or gloss can be evaluated in vitro by measuring color changes in skin coverings, for example by colorimetric testing (Δa) on damaged skin coverings. and ΔE The damaged skin covering is preferably treated with hydrogen peroxide and has been exposed to sunlight, and then treated with the hydrolyzed extract according to the invention. In an alternative embodiment, when in the presence of the hydrolyzed extract according to the invention, preferably under the conditions described in Example 12, Δa The value is relative to Δa measured in the absence of the hydrolyzed extract according to the invention. The value increases by 4%, preferably 6%, and / or, under the conditions of Example 12, for Δa. The calculated percentage efficacy is at least 20%, favorablely at least 30%, particularly at least 80%, and / or ΔE. The value is relative to ΔE measured without treatment with the hydrolyzed extract according to the invention. The value is reduced by 2%, preferably 3%, and / or under the conditions of Example 12 for ΔE. When the calculated percentage efficacy is at least 15%, advantageously at least 20%, and particularly at least 25%, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the color and / or shine of the skin covering.” Advantageously, the skin covering is keratin fibers, preferably hair.

[0081] Biomechanical properties can also be evaluated in vitro by measuring the amount of ATP in the fibroblasts of the hair follicle papilla. In alternative embodiments, the amount of the hydrolyzed extract according to the invention is effective for “maintaining and / or increasing the biomechanical properties of skin covering, preferably keratin fibers, and advantageously hair” when, in the presence of the hydrolyzed extract according to the invention, preferably in the presence of the hydrolyzed extract prepared according to Example 1a, the percentage of papillary fibroblast aggregates under the conditions described in Example 14 is greater than 10%, preferably greater than 20%, more preferably greater than 25%, and advantageously greater than 30% relative to the percentage of papillary fibroblast aggregates measured without treatment with the hydrolyzed extract according to the invention.

[0082] Therefore, the hydrolyzed extract according to the invention is 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 covering and, preferably, hair, and thus can reduce its brittleness, breakage, roughness, and thus improve detangling and / or styling.

[0083] The term "maintaining and / or increasing the biomechanical properties of the skin covering and beneficial hair" means making the skin covering and beneficial hair visually smoother and / or shinier and / or brighter and / or easier to manage and / or less prone to split ends. These properties can be evaluated using imaging techniques, such as video microscopy or electron microscopy, or by sensory testing of hair strands (performed by trained external volunteers to determine their visual or tactile feel), or additionally by evaluation tests conducted by experts (e.g., hairdressers) or trained volunteers, or by self-evaluation using consumer questionnaires. The efficacy of a product in its formulation is evaluated based on, in particular, perceptible visual or tactile quality standards of hair.

[0084] The improved repair and / or biomechanical properties of the hydrolyzed extract according to the invention make it a cosmetic active ingredient for improving the resistance and / or strength and / or volume and / or color and / or shine and / or brightness of skin coverings, particularly keratin fibers, and preferably hair.

[0085] 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 animal-derived enzymes (selected from pepsin, pancreatic-derived enzymes such as trypsin or chymotrypsin, preferably trypsin), plant-derived enzymes (selected from papain, bromelain, fig protease, kiwi protease, preferably papain), or bacterial-derived enzymes (selected from Bacillus licheniformis strains, particularly those sold under the name Alcalase®, or from Bacillus subtilis strains, preferably from Bacillus licheniformis strains sold under the name Alcalase®). In a particularly advantageous embodiment of the invention, the enzymes used are derived from Bacillus licheniformis strains and are particularly sold under the name Alcalase®.

[0086] The amount of milk thistle seed cake according to the invention used for hydrolysis is advantageously between 1% and 20% by weight, more advantageously between 2% and 15%, and even more advantageously between 10% and 15%, relative to the total weight of the solvent and the cake.

[0087] The solvent that can be used for hydrolysis can be an aqueous solvent.

[0088] According to the present invention, the term "aqueous solvent" means a total aqueous solution containing, by weight, more than 60%, advantageously at least 70%, particularly at least 80%, more particularly at least 90%, and particularly at least 95% water relative to the aqueous solution, or even more advantageously, at least free of glycols and especially alcohols. The aqueous solvent does not contain any aqueous solutions, subcritical water, ethanol, isopropanol, or propylene glycol.

[0089] Advantageously, the solvent used is water, which is the only solvent.

[0090] Depending on the optimal pH of the enzyme, hydrolysis can be carried out at pH between 3 and 10, and advantageously at pH between 7 and 9, very advantageously at pH between 7.5 and 8.5, and even more advantageously at pH 7.5.

[0091] 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.

[0092] Hydrolysis can be carried out at temperatures between 30°C and 75°C, advantageously between 50°C and 70°C, particularly between 55°C and 65°C, and preferably at 65°C. It can be carried out over a period of 30 minutes to 24 hours, preferably 45 minutes to 12 hours, more preferably 50 minutes to 6 hours, and very preferably 1 hour to 3 hours.

[0093] The amount of enzyme used for hydrolysis is between 0.1% and 25% by weight relative to the weight of the cake, preferably between 0.5% and 20% by weight, more preferably between 0.75% and 10% by weight, and advantageously between 1% and 5% by weight.

[0094] The enzyme can then be inactivated by heating, advantageously at a temperature of 80°C to 100°C and very advantageously at 90°C, particularly for a period of 10 minutes to 3 hours, preferably 10 minutes to 2 hours, more preferably 20 minutes to 1 hour and 30 minutes. Enzyme inactivation can occur at a pH between 3.5 and 8, preferably between 4 and 7, more preferably between 4 and 6.5, and advantageously at pH 6.0.

[0095] The obtained hydrolyzed extract can then be centrifuged and clarified by continuous filtration down to a porosity of 0.22 µm (filtration threshold).

[0096] Therefore, the hydrolyzed extract according to the invention can be in liquid form, which advantageously contains 1% to 20% by weight, 2% to 10% by weight, and more advantageously between 3% and 6% by weight of the total weight of the hydrolyzed extract in liquid form.

[0097] In alternative embodiments, the aqueous extraction step is performed prior to the hydrolysis step by any method known to those skilled in the art, particularly by maceration and advantageously by extraction using water as the sole solvent. The amount of cake used for aqueous extraction is between 1% and 20% by weight relative to the total weight of the solvent and cake, advantageously between 2% and 15%, and more advantageously between 10% and 15%.

[0098] In this case, 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.

[0099] Aqueous extraction can be carried out at temperatures between 4°C and 300°C, advantageously between 15°C and 100°C, more advantageously between 20°C and 90°C, and very advantageously between 20°C and 70°C. In a particularly advantageous embodiment of the invention, aqueous extraction is carried out at room temperature (i.e., between 20°C and 25°C).

[0100] In a preferred embodiment, the aqueous extraction is carried out at 20°C and pH 12 using a cake amount of 15% by weight relative to the total weight of the cake and water.

[0101] In another embodiment, the aqueous extraction was carried out at 20°C and pH 12 using a cake amount of 12.5% ​​by weight relative to the total weight of the cake and water.

[0102] In an alternative embodiment, the aqueous extraction is performed under subcritical conditions.

[0103] The term "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 liquid but has lower viscosity and surface tension than water at room temperature, increasing the dielectric constant of water. Therefore, the extraction pressure will 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).

[0104] Therefore, under subcritical conditions, extraction is carried out in water, particularly at temperatures ranging from 100°C to 300°C, advantageously from 120°C to 250°C, and more advantageously between 120°C and 180°C. Extraction can be carried out at a single given temperature or at continuously increasing temperatures. In an advantageous embodiment of the invention, extraction will be carried out at a single temperature of 160°C. In alternative embodiments, it will be carried out according to a gradient of three increasing temperatures between 100°C and 200°C (e.g., 120°C, 140°C then 160°C, or 110°C, 130°C then 150°C, or alternatively 120°C, 145°C then 170°C).

[0105] In a preferred embodiment, subcritical extraction was performed at 120°C for 20 minutes at a pressure of 5 MPa (50 bar). The resulting extract was then filtered under hot conditions.

[0106] In an alternative embodiment, subcritical extraction was performed at a pressure of 5 MPa (50 bar) and a temperature of 140°C for 20 minutes. The resulting extract was then filtered under hot conditions.

[0107] In an alternative embodiment, the resulting hydrolyzed extract of milk thistle seed cake in liquid form can be used as a solvent for hydrolyzing new quantities of milk thistle seed cake.

[0108] In an advantageous embodiment, the hydrolyzed extract is obtained by enzymatic digestion as follows: milk thistle seed cake is milled and suspended in water (as the sole solvent) at a concentration of 12.5% ​​of the seed cake relative to the total weight of the cake and solvent, and subjected to hydrolysis at 55°C and pH 8.5 for a period of 1 hour, maintaining the pH during hydrolysis, wherein the weight concentration of a liquid bacterial protease (Alcalase®) from the Bacillus licheniformis strain is 1% relative to the weight of the cake. The enzyme is then inactivated by heating at 90°C and pH 4.5 for 1 hour. The mixture is then cooled under the conditions described in Example 1b), centrifuged, and the supernatant is filtered (filtration threshold: 0.22 µm).

[0109] In a particularly advantageous embodiment of the invention, the hydrolyzed extract is obtained by enzymatic digestion as follows: milk thistle seed cake is milled and suspended in water (as the sole solvent) at a concentration of 15% by weight relative to the total weight of the cake and solvent, and subjected to hydrolysis at 65°C for a period of 2 hours at pH 7.5, maintaining the pH during hydrolysis, wherein the weight concentration of a bacterial protease (Alcalase®) in liquid form from the Bacillus licheniformis strain is 1.7% relative to the weight of the cake. The enzyme is then inactivated by heating at 90°C for 1 hour at pH 6.0. The mixture is cooled to room temperature (between 20°C and 30°C) under the conditions described in Example 1a), centrifuged, and filtered (filtration threshold: 0.22 µm).

[0110] The terms “small peptide” or “low molecular weight peptide” refer to peptides with a weight-average molecular weight of less than 7,000 Daltons, which are analyzed by gel permeation chromatography (such as Superdex® Peptide 10 / 30 HR GE or Superose® 12 10 / 300 GL) and subsequently by a protein molecular calibration column of known molecular weight (gel permeation chromatography (GPC)).

[0111] Advantageously, relative to the total dry mass of the hydrolyzed extract according to the invention, 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, and more preferably between 60% and 70% by weight.

[0112] Advantageously, relative to the total dry mass of the hydrolyzed peptides in the hydrolyzed extract according to the invention, the hydrolyzed extract according to the invention has a dry mass content of peptides with a weight-average molecular weight between 100 Da and 20 kDa, as analyzed by gel permeation chromatography (such as Superose® 12 10 / 300 GL), between 50% and 90% by weight.

[0113] Advantageously, relative to the total dry mass of peptides having a molecular weight between 100 Da and 7000 Da in the hydrolyzed extract according to the invention, the hydrolyzed extract according to the invention has a dry mass content (column sensitivity) of peptides having a weight-average molecular weight between 100 Da and 700 Da, which can be determined by gel permeation analysis on a Superdex® Peptide 10 / 30 HR GE column, between 60% and 90% by weight, advantageously between 70% and 80%, and more advantageously between 75% and 80% by weight.

[0114] Advantageously, relative to the total dry mass of peptides having a molecular weight between 100 Da and 7000 Da in the hydrolyzed extract according to the invention, the hydrolyzed extract according to the invention has, by weight, a dry mass content (column sensitivity) of peptides having a weight-average molecular weight between 100 Da and 2500 Da, which can be analyzed by gel permeation on a Superdex® Peptide 10 / 30 HR GE column, between 70% and 99%, advantageously between 80% and 99%, and more advantageously between 90% and 99%, of peptides having a weight-average molecular weight between 100 Da and 2500 Da.

[0115] Even more advantageously, the hydrolyzed extract according to the invention is not used in combination with retinoids.

[0116] The hydrolyzed extract according to the present invention can be used alone or incorporated into cosmetic compositions.

[0117] In one embodiment, the hydrolyzed extract can then be dried, for example by freeze-drying or atomization, with or without maltodextrin. The hydrolyzed extract is then in powder form.

[0118] According to one embodiment, the hydrolyzed extract according to the invention, particularly the hydrolyzed extract obtained under the conditions described in Examples 1a) and 1b), can advantageously be atomized under the conditions of Example 1c) or 1d) in the presence of maltodextrin at a weight concentration between 1% and 99%, advantageously between 5% and 90%, preferably between 40% and 85%, more preferably between 65% and 85%, and even more preferably between 70% and 80% relative to the total weight of the obtained powder, and the hydrolyzed extract according to the invention at a weight concentration between 1% and 99%, advantageously between 10% and 95%, preferably between 15% and 60%, more preferably between 15% and 25%, and even more preferably between 20% and 30% relative to the total weight of the obtained powder.

[0119] Alternatively, hydrolyzed extracts can be used alone in solid form, as a cosmetic or dermatological ingredient.

[0120] Alternatively, when used alone in liquid form as a cosmetic or dermatological ingredient, it is advantageously dissolved in an aqueous solution containing glycerin, advantageously at a concentration of 60% to 90%, more advantageously 70% to 85%, and very advantageously 80% by weight relative to the total weight of the aqueous solution containing the hydrolyzed extract.

[0121] In alternative embodiments of the invention, the hydrolyzed extract is dissolved and / or diluted in a solvent, particularly a polar solvent (such as water, alcohol, polyol, glycol, such as pentanediol and / or butylene glycol and / or hexanediol and / or octanediol), or a mixture thereof (preferably a water-glycol mixture, more preferably containing glycols selected from hexanediol, propylene glycol, octanediol, and any mixture thereof). Advantageously, the hydrolyzed extract according to the invention is diluted and / or dissolved in an aqueous solution containing hexanediol, particularly containing between 0.1% and 10% by weight of hexanediol relative to the total weight of the cosmetic ingredients, preferably between 0.5% and 5% by weight. Advantageously, the hydrolyzed extract according to the invention is diluted and / or dissolved in an aqueous solution containing octanediol, particularly containing between 0.01% and 5% by weight of octanediol relative to the total weight of the aqueous solution containing the hydrolyzed extract, preferably between 0.1% and 1% by weight. Alternatively, the solution in which the hydrolyzed extract according to the invention is dissolved contains pentanediol and octanediol.

[0122] In particular, the aqueous solution in which the hydrolyzed extract of the present invention is dissolved contains xanthan gum, especially xanthan gum at a weight of 0.01% to 5% relative to the total weight of the aqueous solution, and more particularly xanthan gum at a weight of 0.1% to 1% relative to the total weight of the aqueous solution containing the hydrolyzed extract.

[0123] Hydrolyzed extracts can be used alone as cosmetic ingredients or incorporated into cosmetic compositions containing at least one cosmetically acceptable excipient.

[0124] Within the meaning of this invention, the term "cosmetically acceptable" excipient refers to a cosmetically acceptable, particularly locally acceptable, compound and / or solvent that does not cause unreasonable inflammation or allergic reactions upon contact with the skin, especially the scalp, is non-toxic and not unstable, or its equivalent.

[0125] Within the meaning of this invention, the term "cosmetic composition" refers to a non-therapeutic composition, i.e., a composition intended for the prevention and / or care of skin, especially the scalp and / or skin coverings, especially hair, which dermatologists refer to as "normal," i.e., non-pathological. The term "normal" in relation to skin, scalp, skin coverings, or hair is used herein to mean healthy skin, scalp, skin coverings, or hair as previously defined.

[0126] In a preferred embodiment of the invention, the hydrolyzed extract according to the invention, by weight, is 1 × 10⁻⁶ relative to the total weight of the composition. -4 Between % and 10%, preferably 1 × 10 by weight -3 % to 5%, more preferably 1 × 10 by weight -3 % to 3%, or even more preferably 1 × 10 by weight -3 It is present in the cosmetic composition at a content of % to 1%, or advantageously 0.01% to 1% by weight.

[0127] Therefore, the composition can be used to maintain and / or increase the biomechanical properties of skin coverings, keratin fibers, and preferably hair.

[0128] Cosmetic compositions according to the invention can be in forms conventionally used for topical application to the skin or scalp and / or skin coverings, preferably scalp and / or skin coverings, such as liquid or solid forms or even pressurized liquid forms. They can be formulated as aqueous or oily solutions, aqueous creams or gels or oily gels, especially in cans or tubes, particularly as shower gels, shampoos, conditioners, emulsions, oils, lotions, hydrogels, microemulsions or nanoemulsions (especially oil-in-water or water-in-oil or multi- or silicone emulsions), serums, lotions (especially in glass or plastic bottles or metering bottles, aerosols or sprayers, vials), liquid soaps, pastes, dermatological sticks, ointments, foams, masks, hairsprays, patches, nail polishes, and anhydrous products (which are preferably liquid, paste or solid, for example in sticks, especially rods, or in powder form). It can be a cosmetic product, particularly a cosmetic product for eyelashes or eyebrows, such as mascara or a pencil, or a makeup remover for eye care and beauty and / or a product for nail care and beauty and / or a product for beard care and / or beauty. Specifically, the cosmetic composition is selected from the group consisting of: serums, lotions, creams, shampoos, conditioners, oils, emulsions, ointments, pastes, foams, lotions, hydrogels, shower gels, face masks, hairsprays, sprays, waxes, mascaras, makeup pencils, or nail polish, preferably shampoos, conditioners, or lotions.

[0129] Preferably, the hydrolyzed extract according to the invention is suitable for formulating "neutral" compositions that are mild on skin coverings, and advantageously on keratin fibers, especially hair fibers. The hydrolyzed extract according to the invention is also suitable for use in cationic formulations containing surfactants.

[0130] Cosmetic compositions according to the invention may contain any suitable solvent and / or any suitable medium and / or any suitable excipient, optionally in combination with other compounds of interest. They may, in particular, contain cosmetically acceptable excipients selected from surfactants, preservatives, buffers, swelling agents, chelating agents, biocides, denaturants, sunscreens, pH adjusters, reducing agents, stabilizers, emulsifiers, thickeners, gelling agents, film-forming polymers, solvents, fillers, bactericides, odor absorbers, shine removers, conditioning agents, texturing agents, gloss agents, pigments, dyes, fragrances, chemical or mineral sunscreens, trace elements, and essential oils. These combinations are also covered by the invention.

[0131] Cosmetic compositions may contain other cosmetic agents having the same properties as the hydrolyzed extract according to the invention and potentially inducing a synergistic effect with said hydrolyzed extract, or cosmetic agents with complementary effects, such as agents for combating hair loss, hair protectants, soothing agents, anti-aging agents, or anti-pollution agents. As an active agent for combating hair loss, combinations of the following may be mentioned: sulfonyl peptides, amino acids, amino sugars, B vitamins, zinc, and extracts of ginseng and burdock root sold by the applicant under the name Trichigen™ LS 8960; or hair protectants, such as extracts of litchi peel sold by the applicant under the name Litchiderm™; and soothing and antipruritic agents, such as rapeseed phytosterols sold by the applicant under the name Phytosoothe™ LS9766.

[0132] Other active agents may be present in the composition, such as an antioxidant active agent, specifically for hair care, an extract of *Cassia tora* leaf marketed under the name DN-Age™; a combination of an anti-glycation agent, an extract of *Salvia miltiorrhiza* marketed under the name CollRepair™, and niacinamide; or active agents that improve skin firmness and thus scalp firmness, such as a synthetic tetrapeptide marketed under the name Dermican™; an extract of *Abelmoschus manihot* marketed under the name Linefactor™; a purified pea extract marketed under the name Proteasyl™; an extract of *Manilkara multinervis* marketed under the name Elestan™; an extract of *Melia azedarach* marketed under the name Collalift™ 18; an extract of *Argassential* pulp marketed by the applicant under the name Argassential™; an extract of *Schisandra chinensis* marketed under the name Sqisandryl™; an extract of *Caesalpinia falcatum* marketed under the name Eperuline™; and an extract of *Orthosiphon staminus* marketed by the applicant under the name MAT-XS™ Bright. The combination of these active agents is particularly effective in strengthening hair follicles and reducing hair loss. The hydrolyzed extract of this invention can also be combined with an extract of the seeds of the plant Rambuvital, sold by the applicant under the name Rambuvital™, for its hair-protecting properties, especially against pollutants. The hydrolyzed extract can also be used in combination with a hydrolyzed extract of sea buckthorn seed cake.

[0133] Advantageously, the cosmetic compositions according to the invention do not contain any retinoids.

[0134] A third aspect of the invention relates to a non-therapeutic cosmetic care method comprising the topical application of a hydrolyzed extract according to the invention or a cosmetic composition containing the same to maintain and / or increase skin coverage, beneficial keratin fibers, and preferably the biomechanical properties of hair.

[0135] In an advantageous embodiment of the invention, the cosmetic care method comprises the following: topically applying the hydrolyzed extract of the invention or a cosmetic composition containing the present invention to all or part of the skin of the body and / or face, including skin coverings, particularly selected from the scalp, legs, thighs, arms, stomach, neckline, neck, all or part of the face, forehead, chin, perilips, perieyes, and the area of ​​the face known as the "T-zone," and advantageously all or part of the scalp and / or skin coverings, advantageously nails, hair, body hair (especially beard, eyelashes and / or eyebrows), even more advantageously hair, preferably all or part of keratin fibers, even more preferably all or part of hair.

[0136] Therefore, beauty care methods allow for the maintenance and / or increase of skin coverage, beneficial keratin fibers, more favorable hair resistance and / or strength and / or volume and / or color and / or shine and / or styling (i.e., non-brittle appearance), and / or smoothness.

[0137] Another subject also relates to a cosmetic treatment method for maintaining and / or increasing the biomechanical properties of skin covering, preferably keratin fibers, and hair, and the method includes the following steps:

[0138] - Identify areas of body and / or facial skin (including skin coverings) and / or areas of skin coverings in individuals who do not have a pathology requiring therapeutic treatment, particularly dermatopathology, for which it is desirable to maintain and / or increase the skin coverings, preferably keratin fibers, advantageously hair, and / or the biomechanical properties required thereto.

[0139] - Apply topically, according to the invention, the hydrolyzed extract or composition comprising it, to areas of the body and / or facial skin (including skin coverings) and / or areas of the skin coverings, particularly at a weight of 1 × 10⁻⁶ relative to the total weight of the composition. -4 Between % and 10%, preferably 1 × 10 by weight -3 % to 5%, more preferably 1 × 10 by weight -3 % to 3%, or even more preferably 1 × 10 by weight -3 % to 1%, advantageously 0.01% to 1% by weight.

[0140] Examples form part of the invention, and any feature that appears novel relative to the prior art when considered in its entirety (including the examples) forms part of the invention. Therefore, each example has a general scope.

[0141] Unless otherwise stated, temperature is in degrees Celsius and pressure is in atmospheres. Detailed Implementation

[0142] Example

[0143] Example 1: Different methods for preparing hydrolyzed extracts of milk thistle seed cake according to the present invention

[0144] The biscuits used in the following examples contain less than 8% lipids by weight relative to the total weight of the biscuits.

[0145] Example 1a: Enzymatic hydrolysis – hydrolyzed extract 1

[0146] Milk thistle seed cake was ground and suspended in water (as the sole solvent) at a concentration of 15% by weight relative to the total weight of the cake and solvent. Hydrolysis was then carried out at 65°C and pH 7.5 for 2 hours, maintaining the pH during hydrolysis. The concentration of a liquid bacterial protease (Alcalase®) from the *Bacillus licheniformis* strain was 1.7% by weight relative to the cake weight. The enzyme was then inactivated by heating at 90°C and pH 6.0 for 1 hour. The mixture was cooled to room temperature, centrifuged, and filtered (filtration threshold: 0.22 µm). A liquid product containing 4.14% ± 0.3% by weight relative to the liquid product was obtained.

[0147] Example 1b: Enzymatic hydrolysis – hydrolyzed extract 2

[0148] Milk thistle seed cake was ground and suspended in water (as the sole solvent) at a concentration of 12.5% ​​of the seed cake relative to the total weight of the cake and solvent. Hydrolysis was then carried out at 55°C and pH 8.5 for 1 hour, maintaining the pH during hydrolysis. The concentration of a liquid bacterial protease (Alcalase®) from a Bacillus licheniformis strain was 1% by weight relative to the weight of the cake. The enzyme was then inactivated by heating at 90°C and pH 4.5 for 1 hour. The mixture was then cooled, centrifuged, and the supernatant was filtered (filtration threshold: 0.22 µm). A liquid product containing 3.26% dry matter relative to the weight of the liquid product was obtained.

[0149] Example 1c: Hydrolyzed extract 1 in cosmetic form (containing maltodextrin)

[0150] The extract obtained in Example 1a) was atomized in the presence of maltodextrin (at a final maltodextrin amount of 75% by weight relative to the total weight of the final atomized extract). Thus, an extract in powder form was obtained.

[0151] Example 1d: Hydrolyzed extract 2 in cosmetic form (containing maltodextrin)

[0152] The extract obtained in Example 1b) was atomized in the presence of maltodextrin (at a final amount of 75% (w / w) by weight relative to the total weight of the final atomized extract). Thus, an extract in powder form was obtained.

[0153] Example 1e: Enzymatic hydrolysis using hydrolyzed extract 1 according to Example 1a

[0154] Milk thistle seed cake was ground and suspended in the hydrolyzed extract (sole solvent) obtained in Example 1a) at a concentration of 15% by weight of the cake relative to the total weight of the cake and solvent. The resulting mixture was subjected to hydrolysis at 65°C and pH 7.5 for 2 hours, maintaining the pH during hydrolysis. The concentration of the liquid bacterial protein (Alcalase®) derived from the Bacillus licheniformis strain was 1.7% by weight of the cake. The enzyme was then inactivated by heating at 90°C and pH 6.0 for 1 hour. The mixture was cooled to room temperature, centrifuged, and filtered (filtration threshold: 0.22 µm). A liquid product containing 6.1% by weight of dry matter relative to the weight of the liquid product was obtained.

[0155] Example 2. Peptide equivalent content of hydrolyzed extracts in cosmetic ingredient form according to Example 1a and Example 1c. Analysis of quantity and peptide molecular weight distribution

[0156] a) Determination of total protein

[0157] The purpose of this study is to quantify the total peptide equivalent content of the hydrolyzed extract according to Example 1a of the present invention.

[0158] Materials and methods: The peptide equivalent content of the hydrolyzed extract was estimated by total nitrogen determination (Kjeldahl method - AOAC, Association of Official Analytical Chemists Official Methods of Analysis, AOAC, Washington, D.C., 2000) and the obtained value was multiplied by a factor of 6.25 (N × 6.25). The results are summarized in Table 1.

[0159] [Table 1]

[0160]

[0161] result:The dry matter content obtained after extraction and hydrolysis was between 3.84% and 4.44%. Total peptides accounted for approximately 60% to 66.2% of the dry matter of the hydrolyzed extract according to Example 1a (Table 1).

[0162] in conclusion: The hydrolyzed extract of milk thistle seed cake according to the present invention is rich in plant peptides (between 60% and 66.2% by weight relative to the total weight of dry matter).

[0163] b) Analysis of the average molecular weight distribution of hydrolyzed peptides

[0164] Molecular weight distribution of hydrolyzed extracts

[0165] Purpose: To characterize the weight-average molecular weight of the peptide compounds in the hydrolyzed extracts according to the present invention, two hydrolyzed extracts according to Example 1a were studied and then classified into three categories: less than 100 Da, 100 Da to 20 kDa (inclusive), and greater than 20 kDa.

[0166] Materials and methods: The weight-average molecular weight distribution of peptides from the hydrolyzed extract in cosmetic form according to Example 1a was analyzed by gel permeation chromatography on a Superose® 12 10 / 300 GL column (GE Healthcare Life Sciences). The weight-average molecular weight was determined after column calibration with protein molecules of known weight-average molecular weight. The results are summarized in Table 2.

[0167] [Table 2]

[0168]

[0169] result: The peptides of the hydrolyzed extract of milk thistle seed cake according to the present invention are mainly peptide compounds with a weight-average molecular weight between 100 Da and 20 kDa (Table 2).

[0170] Molecular weight distribution between 100 Da and 7000 Da

[0171] Purpose: The aim of this study was to investigate the weight-average molecular weight distribution of peptides contained in the hydrolyzed extract of milk thistle seed cake obtained according to Example 1c, within the range of 100 Da to 7000 Da. The weight-average molecular weights were then classified into three categories: two extreme categories combining the minimum molecular weight of 100 Da < MW < 700 Da and the maximum molecular weight of 2500 Da < MW < 7000 Da.

[0172] Materials and methods:The weight-average molecular weight distribution of peptides from the hydrolyzed extract of milk thistle seed cake obtained according to Example 1c was analyzed by gel permeation chromatography on a Superdex® Peptide 10 / 30 HR GE column (GE Healthcare Life Sciences). Fractionation was performed from 100 Da to 7000 Da, and UV detection was performed at 214 nm. The weight-average molecular weight was determined after column calibration with protein molecules of known weight-average molecular weight. The results are summarized in Table 3.

[0173] [Table 3]

[0174]

[0175] result: The hydrolyzed extract of milk thistle seed cake according to the present invention mainly comprises (98.03%) protein compounds with low weight-average molecular weight in the range of 100 Da to 2500 Da (Table 3).

[0176] Example 3: In vitro effects of hydrolyzed extracts from Example 1b on the biomechanical properties of skin coverings - from Radical scavenging activity (DPPH)

[0177] The aim of this study was to measure the free radical activity of the hydrolyzed extract according to Example 1b using the DPPH method.

[0178] Materials and methods:

[0179] Diphenyl-1-picrylhydrazyl (DPPH°) is an oxidant in its electron-deficient free radical form. In this form, the deep purple compound absorbs wavelengths between 510 nm and 530 nm. In the presence of an electron-donating reducing compound, diphenyl-2-picrylhydrazyl forms 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 was used to select plant extracts with free radical scavenging 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. [International Journal of Cosmetic Science] Dec. 1997; 19(6):299-307).

[0180] The hydrolyzed extract of milk thistle seed cake according to Example 1b was mixed in distilled water at 5% (w / v). 100 µl of this solution was tested in the presence of an equal volume of 90 µM diphenyl-1-picrylhydrazine solution (DPPH°, Sigma – 3.55 mg DPPH powder in 100 mL ethanol). After incubation at room temperature and in the dark for 30 min, absorbance was recorded at 530 nm using a spectrometer (Victor V, Perkin Elmer). The results are expressed as a percentage of DPPH° radical inhibition relative to the negative control.

[0181] result: These are summarized in Table 4. The hydrolyzed extract of milk thistle seed cake according to the present invention retains up to 95% of the DPPH oxidation (Table 4).

[0182] [Table 4]

[0183]

[0184] in conclusion:The hydrolyzed extract of milk thistle seed cake according to the present invention has strong free radical activity and therefore has an effect on maintaining and / or increasing biomechanical properties.

[0185] Example 4: In vitro biomechanical properties of the hydrolyzed extract of the seed cake according to the present invention on skin coverings Effects - Antioxidant / Transition Metal (Fe) Chelating Activity

[0186] Glycosylation can occur chemically between reducing sugars and proteins. The presence of oxidants and metals catalyzes this reaction and the formation of AGEs (advanced glycation end products).

[0187] The chelating and antioxidant activities of the hydrolyzed extract of milk thistle cake according to the present invention were evaluated in an in vitro model in the presence of albumin, iron, and the oxidant hydrogen peroxide. Oxidation reactions were evaluated by measuring tryptophan degradation, tyrosine bridging, and the formation of glycosylation products (including pentasaccharides).

[0188] Materials and methods:

[0189] A reagent containing 1.5% (w / v) albumin, 2 mM iron, 5 mM EDTA, and 25 mM hydrogen peroxide (H₂O₂) was brought into contact with, or not into contact with, a hydrolyzed extract of milk thistle cake according to Example 1a at a concentration of 0.003% or 0.01% by weight. Albumin was oxidized in the presence of H₂O₂. The oxidation reaction was catalyzed by iron. A control of the catalytic oxidation reaction was carried out in the absence of iron. The mixture was incubated at 37°C for 1 day.

[0190] The amounts of tryptophan, dityrosine, pentosaccharide, and fluorescent glycosylation products were measured by fluorescence using a 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 pentosaccharide; 370 nm / 440 nm for glycosylation end products).

[0191] The statistical significance threshold was set at 5% (p < 0.05).

[0192] The protection index is calculated according to the following formula: (iron-containing control - treated with the hydrolyzed extract according to the invention and iron) / (iron-containing control - iron-free control) × 100.

[0193] The dried extract according to Example 1a was tested to obtain 100% dry extract content.

[0194] [Table 5]

[0195]

[0196] The presence of iron catalyzes the oxidation reaction, which can be observed by the increase in the content of undegraded tryptophan in the iron-free albumin control (Table 5). In the presence of 0.003% and 0.01% (by weight) of the hydrolyzed extract of milk thistle seed cake according to Example 1a, the degradation of tryptophan was significantly reduced, with 18% and 28% of the undegraded tryptophan residues recovered, respectively. The protection index of the hydrolyzed extract of milk thistle seed cake tested at 0.01% (by weight) was 38%.

[0197] [Table 6]

[0198]

[0199] The presence of iron catalyzes the formation of dityrosine in albumin in the presence of H2O2 (Table 6). Hydrolyzed extracts of milk thistle seed cake according to Example 1a at 0.003% and 0.01% by weight reduced dityrosine formation by 16% and 28%, respectively. The protection index of the hydrolyzed extract of milk thistle seed cake tested at 0.01% by weight was 67%.

[0200] [Table 7]

[0201]

[0202] The presence of iron also catalyzes the formation of pentosaccharides in the presence of H2O2 (Table 7). A 0.01% by weight hydrolyzed extract of milk thistle seed cake according to Example 1a reduced pentosaccharide formation by 27%. The protection index of the hydrolyzed extract of milk thistle seed cake tested at 0.01% by weight was 61%.

[0203] [Table 8]

[0204]

[0205] The presence of iron catalyzes the oxidation of albumin in the presence of H2O2 and is accompanied by the formation of fluorescent glycosylation products (Table 8). Hydrolyzed extracts of milk thistle cake according to Example 1a at 0.003% and 0.01% by weight reduced the formation of these glycosylation products by 9% and 14%, respectively. The protection indices of the hydrolyzed extracts of milk thistle cake tested at 0.003% and 0.01% by weight were 18% and 27%, respectively.

[0206] in conclusion: Hydrolyzed extracts of milk thistle seed cake reduce iron-catalyzed protein oxidation and the formation of glycosylation products, and thus maintain biomechanical properties.

[0207] Example 5: In vitro biomechanical properties of the hydrolyzed extract of the seed cake according to the present invention on skin coverings. Effects - Antioxidant / transition metal (Cu) chelating activity

[0208] The aim of this study was to evaluate the antioxidant / metal chelating activity of the hydrolyzed extract of milk thistle cake according to the present invention in an in vitro assay in the presence of lipoproteins and copper.

[0209] Materials and methods

[0210] 200 µM copper was placed in contact with 100 µg / mL low-density lipoprotein (LDL) in a phosphate-buffered saline solution. 0.048% by weight of a hydrolyzed extract of milk thistle seed cake according to Example 1a was added to the solution, or not. The reaction mixture was incubated at 37°C for 48 hours.

[0211] After adding 2% by volume of thiobarbituric acid (TBA) relative to the total volume of the reaction mixture, the level of lipid oxidation was evaluated by quantitative malondialdehyde (MDA). Following excitation at 532 nm, the fluorescence level was quantified spectrophotometrically at emission wavelength of 560 nm using a ThermoScientific Varioskan® Flash device.

[0212] Schiff base content was evaluated using a Thermo Scientific Varioskan® Flash device via spectroscopic methods (excitation wavelength 370 nm / emission wavelength 440 nm).

[0213] The statistical significance threshold was set at 5% (p < 0.05).

[0214] The protection index is calculated according to the following formula: (using the hydrolyzed extract according to the invention and the LDL + copper treatment - LDL + copper control) / (control without copper - LDL + copper control) × 100.

[0215] result: The results are summarized in Table 9 below.

[0216] [Table 9]

[0217]

[0218] The presence of copper enhances lipid oxidation as determined by malondialdehyde (MDA). A hydrolyzed extract of milk thistle seed cake according to Example 1a, used at 0.048% by weight, significantly reduced MDA formation by 78% (Table 9). The protection index of the hydrolyzed extract of milk thistle seed cake according to Example 1a, tested at 0.048% by weight, was 97.5%.

[0219] in conclusion:The hydrolyzed extract of milk thistle seed cake according to the present invention reduces copper-catalyzed lipid oxidation and thus allows for the maintenance and / or increase of biomechanical properties.

[0220] Example 6: In vitro biomechanical properties of the hydrolyzed extract of the seed cake according to the present invention on skin coverings Effects – Anti-glycation activity of hair fiber proteins

[0221] Purpose: The aim of this study was to measure the anti-glycation effect of hydrolyzed extracts of milk thistle seed cake according to Example 1a on hair fiber proteins incubated in glucose solution.

[0222] Materials and methods:

[0223] Standardized, clean hair strands were applied to a solution containing or without the hydrolyzed extract according to Example 1a at a concentration of 1 mol / L. -1 The protein was incubated in (M) phosphate-buffered glucose solution at pH 7.4 and 50°C for 2 days, and dried to obtain a 100% dry extract with a content of 0.01% by weight. The hair strands were then rinsed, wrung out, combed, separated, and cut into 1 cm long pieces. The protein was extracted in 1N sodium hydroxide (NaOH) solution with gentle stirring at room temperature for 24 hours.

[0224] The amounts of dityrosine and pentosan (glycosylated products) were measured by fluorescence using a Thermo Scientific Varioskan® Flash device (excitation / emission wavelengths: 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosan).

[0225] The protection index is calculated according to the following formula: (glycosylated control – glycosylated + treated with the extract according to the invention) / (non-glycosylated control – glycosylated control) × 100.

[0226] The statistical significance threshold was set at 5% (p < 0.05).

[0227] result: They are shown in Tables 10 and 11 below.

[0228] [Table 10]

[0229]

[0230] Incubation of fibers in glucose solution increases the dityrosine content in hair proteins. A hydrolyzed extract of milk thistle seed cake according to Example 1a, used at 0.01% by weight, significantly reduced dityrosine formation by 16% (Table 10). The protective index against dityrosine production of the hydrolyzed extract according to Example 1a, used at 0.01% by weight, was 57%.

[0231] [Table 11]

[0232]

[0233] Incubation of fibers in glucose solution increases the pentosin content in hair proteins. A hydrolyzed extract of milk thistle seed cake according to Example 1a, used at 0.04% by weight, significantly reduced pentosin formation by 17% (Table 11). The protective index against dityrosine production of the hydrolyzed extract according to Example 1a, used at 0.04% by weight, was 59%.

[0234] in conclusion: The hydrolyzed extract of milk thistle seed cake according to the present invention reduces the formation of glycosylation products in hair fibers in the presence of glucose and thus allows for the maintenance and / or enhancement of the biomechanical properties of hair fibers.

[0235] Example 7: In vitro effects of the hydrolyzed extract of the present invention on the biomechanical properties of skin coverings – on Antioxidant / carbonylation activity of keratin

[0236] Purpose: The aim of this study is to measure the protective effect of the hydrolyzed extract of milk thistle seed cake according to the present invention against the oxidation and, in particular, carbonylation of keratin and keratin-associated proteins (KAP).

[0237] Materials and methods:

[0238] Preparation of hair strands: Hair strands from Caucasian donors were used for measurement. Hair was treated with 0.05% (w / v) of hydrolyzed extract of milk thistle seed cake according to Example 1a in distilled water by contact for 10 minutes, followed by washing and drying. Urban particles (fine particles) were applied to the hair fibers (PM; ERM-CZ100 fine dust, particulate matter class 10 (PM10), 170 μg / cm² of polycyclic aromatic hydrocarbons) and the hair was immediately exposed to UVA irradiation (84 J / cm², exposure for 6 hours, LED light source, peak emission at 365 nm) (stressed hair). Hair treated with water or with 0.2% (w / v) N-acetylcysteine ​​(antioxidant – positive control) and exposed or not exposed to UVA and particles served as controls.

[0239] Hair samples from each experimental condition were frozen in liquid nitrogen and kept at -80°C until analysis.

[0240] a) Quantitative carbonylation of proteins using Western blotting

[0241] Materials and methods

[0242] Under different conditions, the carbonylated proteins in hair follicles were analyzed and quantified using Western blotting.

[0243] Total protein (keratin and keratin-associated proteins) was extracted from each hair shaft using methods conventionally known to those skilled in the art. Total protein quantification was performed using the Bradford method. Carbonylated proteins were labeled using specific fluorescent probes (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 signals were quantified by acquiring images of the gels using the Thermo Fisher Scientific iBright software system. Total protein was also stained in the gels using the iBright system (Thermo Fisher Scientific) for normalization. For each sample, a carbonyl score (carbonylated protein / total protein) representing a quantitative measure of carbonylated protein was obtained.

[0244] Carbonyl score (sample X) = Fluorescence value of oxidized protein (sample X) / Fluorescence value of total protein (sample X)

[0245] Calculate the protection index (%) using the following formula:

[0246] Protection index (%) = (Carbonyl score level (stress) − Carbonyl score level (treated with the extract according to the invention)) / (Carbonyl score level (stress) − Carbonyl score level (control)) × 100

[0247] For reference, the control group was considered to have the greatest efficacy (100%) and the stress group had the least efficacy (0%).

[0248] The statistical significance threshold was set at 5% (p < 0.05).

[0249] result: The results are summarized in Table 12 below.

[0250] [Table 12]

[0251]

[0252] A significant increase in protein carbonylation was observed during exposure to stress caused by UVA and pollution (UVA 84 J / cm² and urban PM10 particles) (Table 12).

[0253] A 0.05% (w / v) hydrolyzed extract of milk thistle seed cake according to Example 1a significantly reduced the level of carbonylated proteins in hair fibers by 25% (p < 0.01). This corresponds to a protection index of >100%.

[0254] b) Quantitative analysis of carbonylated proteins using image analysis

[0255] Materials and methods:

[0256] In-situ densitometry analysis of carbonylation signals was performed by analyzing images obtained using Thermo Fisher Scientific's iBright software and processed using Rasband's ImageJ software. Oxidation levels for each experimental group in the stratum corneum and cortex regions were expressed as mean (UFR) ± standard deviation relative to the mean (Tables 13 and 14).

[0257] The statistical significance threshold was set at 5% (p < 0.05).

[0258] [Table 13]

[0259]

[0260] result: UVA and urban particles at 84 J / cm² increased the amount of carbonylated proteins in the cortex and stratum corneum.

[0261] [Table 14]

[0262]

[0263] A 0.05% (w / v) hydrolyzed extract of milk thistle seed cake according to Example 1a significantly reduced the content of carbonylated proteins in the stratum corneum by 28% (p < 0.01) and cortex by 31% (p < 0.001), respectively, thereby maintaining and / or increasing the biomechanical properties of hair. This corresponds to a protection index of 49% for the stratum corneum and 59% for the cortex.

[0264] c) Protection of the structural integrity of hair

[0265] Purpose:The structural state of keratin proteins in the hair shaft obtained according to the above method was evaluated using optical measurements with an XPolar® microscope (KAMAX). The birefringence properties of hair fibers depend on the structural integrity of keratin.

[0266] Hair is composed of fibrous keratin proteins. The crystalline structure of keratin fibers gives hair the ability to alter the polarization of light passing through it; this property is known as birefringence. Birefringence depends on hair thickness and the crystalline state of the keratin. A decrease in the birefringence parameter (K-index) indicates structural damage to keratin. K-index values ​​associated with the birefringence of hair fibers can be determined and compared between damaged and undamaged hair, and between treated and untreated hair.

[0267] Materials and methods:

[0268] Hair segments of 1 cm length (n = 30 for each condition) obtained according to the above scheme were transferred to compatible slides for birefringence analysis using the XPolar technique. This is an imaging device integrated into the microscope, allowing observation and measurement of the polarization properties of the sample under study. First, an image of the reflectance intensity was captured to measure the hair thickness. Then, the sample was illuminated with polarized light to measure Kmax. Assuming the hair has a cylindrical geometry, the birefringence value (K index) can be estimated using an abacus from the Kmax value and the hair diameter.

[0269] The value of improvement in hair structure is obtained by using the damaged group and the untreated group as references according to the following formula: (stressed hair treated with the hydrolyzed extract according to the invention – stressed hair) / (healthy hair – stressed hair) × 100.

[0270] [Table 15]

[0271]

[0272] result: Oxidative stress causes structural changes in hair fibers. A decrease in the K-index was observed in damaged hair compared to undamaged control hair (Table 15). This corresponds to the degradation of keratin structure through oxidation of keratin under exposure to UVA and urban particles.

[0273] Hair fibers treated with 0.05% (w / v) of the hydrolyzed extract of milk thistle seed cake according to Example 1a in distilled water showed a significant increase in the average birefringence value relative to damaged hair fibers (Table 15). The hydrolyzed extract of milk thistle seed cake according to Example 1a allowed for a 14% increase in birefringence (p < 0.05). This corresponds to a 59% improvement in hair structure relative to damaged hair (improvement index), and therefore corresponds to an improvement and / or maintenance of the biomechanical properties of the hair fibers.

[0274] d) Overall hair protection score

[0275] The Hair Protection Score (HPS) is a comprehensive index used to classify the efficacy of hair care products. Score values ​​range from: HPS 15; HPS 30; HPS 50; and HPS 50+. HPS values ​​are obtained using a specific algorithm that integrates early molecular events to evaluate the protection of hair fibers from daily damage. The HPS of a product is obtained by integrating molecular damage assessed by evaluating protein carbonylation and structural damage assessed by evaluating hair birefringence, as described in Example 7, thus supporting effective protection of hair fibers from damage caused by pollution (particulate matter and UVA radiation). An internal reference (N-acetylcysteine: HPS = 50) is used as a control.

[0276] The hydrolyzed extract of milk thistle seed cake from Example 1a showed a hair protection score of 50+. This score is higher than that of N-acetylcysteine.

[0277] statistics: All data were analyzed using GraphPad Prism 9 (Insightful Science USA) or SigmaPlot (Systat Software Inv. USA). Statistical analyses were performed using bivariate Student's t-tests or ANOVA comparisons between the affected group and the condition. The statistical significance threshold was set at 5% (p < 0.05).

[0278] in conclusion: The hydrolyzed extract of milk thistle seed cake according to the present invention protects keratin and keratin-related proteins from carbonylation induced by hair fiber exposure to UV and pollutants, thereby maintaining their structural integrity and biomechanical properties.

[0279] Example 8: The hydrolyzed extract according to the present invention penetrates into the hair fiber (cortex) (FT-IR) (in vitro).

[0280] Purpose: Vibrational spectroscopy techniques such as Raman infrared spectroscopy and Fourier transform infrared spectroscopy (FT-IR) are increasingly used in cosmetic science to study changes in the chemical composition and keratin structure of hair under different environments and treatments. These techniques are non-destructive and do not require sample labeling. FT-IR is particularly advantageous for analyzing colored hair because it experiences less strong absorption by melanin under visible excitation. The aim of this study is to evaluate the distribution of the components of the hydrolyzed extract according to the present invention in hair fibers and to investigate its effect on the infrared profile of keratin.

[0281] Materials and methods:

[0282] Hair preparation and treatment: Testing was performed on three strands of Caucasian hair bleached with persulfate (ultra-bleached). Prior to testing, the strands were normalized (2 g / 2 g / 15 cm), cleaned, rinsed, and dried at room temperature. A braid of healthy Caucasian hair (natural dark brown hair) was used as a control for natural, undamaged hair. The strands were completely covered with the hydrolyzed extract according to Example 1a, mixed in distilled water at 5% w / v or with water as a placebo. Excess product was removed from the strands by rubbing each strand between a gloved thumb and a gloved forefinger. The hair was cut into 10 μm sections for infrared imaging.

[0283] Infrared spectroscopy: Characterizing the vibrational spectrum of the product by coupling infrared spectroscopy with a microscope. Collect samples from 800-4000 cm⁻¹. -1 The image between them has a pixel size of 6.25 × 6.25 μm. 2 Preprocessing was performed to clean the spectrum. A fitting method was then used to trace the product's spectrum within the hair fibers.

[0284] result: The hydrolyzed extracts from Example 1a were measured at 993, 1053, 1110, 1403, and 1597 cm⁻¹. -1 These features exhibit specific spectral bands that differ from the spectrum of keratin. These spectral markers are used to identify hydrolyzed extracts within hair fibers. The hydrolyzed extracts were observed to be primarily concentrated in the hair cuticle and diffused within the cortex of the fiber.

[0285] Example 9: Effect of the hydrolyzed extract of the present invention on the biomechanical properties of skin coverings - cysteine Molecular recombination and stabilization of hair fibers by acid groups (FT-IR) (in vitro)

[0286] Purpose: The aim was to study the vibrational spectra of keratin more precisely, whether or not it was damaged, and whether or not it was treated with the hydrolyzed extract of milk thistle seed cake according to Example 1a. These analyses demonstrated the interaction of the hydrolyzed extract according to the invention with damaged keratin in the hair cortex, and in particular its overall stabilizing effect, especially its stabilizing effect on sulfonyl alanine type groups and their derivatives.

[0287] Materials and methods

[0288] Strands of brown Caucasian hair were bleached and oxidized, 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.

[0289] Comparative analysis of vibrational spectra of keratin from undamaged hair and from bleached oxidized hair treated or untreated with extracts according to Example 1a mixed in distilled water at 5% w / v using a "fit" analysis: The method of comparing a given infrared spectrum of the control with the infrared spectrum of the product was programmed using the Matlab environment. Variations in the keratin signal were processed using hierarchical clustering analysis to determine the similarity between the spectra. 1174 cm⁻¹ was determined. -1 and 1116 cm -1 The area under the absorption band at that point.

[0290] When these values ​​are compared with each other, the statistical significance threshold is set at 5% (p < 0.05).

[0291] result:

[0292] Oxidative damage caused by chemical bleaching of hair shows the effects on the chemical and structural composition of hair, with observable changes in vibrational spectral curves.

[0293] Hierarchical cluster analysis showed that damaged hair and native hair treated with the hydrolyzed extract of milk thistle seed cake according to Example 1a had spectral profiles more closely resembling those of damaged hair. These results indicate that the hydrolyzed extract of milk thistle seed cake according to the invention interacts with and allows the damaged keratin in the fibrous cortex to stabilize.

[0294] The spectral bands involving the integrity of cysteine ​​bonds were then studied in more detail. These were modified under conditions of chemical oxidation damage, relative to healthy, undamaged hair.

[0295] Specifically, compared to healthy hair, in hair oxidized by bleaching, the values ​​at 840, 1116, 1150, and 1174 cm were [not specified]. -1 The intensity of the spectral bands at the point (referring to the stretching of NH, SO2 and CO outside the bending plane, and the asymmetric stretching of SO3, respectively) increases.

[0296] It has been demonstrated that the hydrolyzed extract according to the present invention can reduce these band strengths to the band strength of the original hair.

[0297] Finally, it was demonstrated that the hydrolyzed extract of milk thistle seed cake according to the present invention can reduce these band strengths to a level closer to that of healthy hair (Table 16).

[0298] In particular, oxidative damage acts on SO, SO2, NH, and CO bonds (the strength of these bonds increases significantly). These groups are generated through oxidation of the groups and cleavage of cysteine ​​disulfide bonds. The hydrolyzed extract of milk thistle seed cake according to the invention shows a significant reduction in the strength of these SO, SO2, NH, and CO bonds.

[0299] The protection index is calculated according to the following formula: (glycosylated control – glycosylated + treated with the extract according to the invention) / (non-glycosylated control – glycosylated control) × 100.

[0300] [Table 16]

[0301]

[0302] in conclusion: The hydrolyzed extract of milk thistle seed cake according to the invention is capable of interacting with and stabilizing the molecular components of keratin and keratin-associated proteins in hair fibers damaged by chemical oxidation. The hydrolyzed extract of milk thistle seed cake according to the invention allows for structural restoration and thus allows for the repair and / or maintenance and / or increase of biomechanical properties (especially strength, plasticity, resistance, and / or flexibility between 14% and 98%), depending on 840 and 1174 cm. -1 The chemical groups between them.

[0303] Example 10: In vitro effects of the hydrolyzed extract of the present invention on the repair of hair fibers

[0304] The purpose of this study is to measure the repair effect of the hydrolyzed extract of milk thistle seed cake according to the present invention on hair fibers damaged by hydrogen peroxide by calorimetry (DSC = differential scanning calorimetry).

[0305] Materials and methods:

[0306] Strands of dark brown Caucasian hair (1 g; 12 cm) were calibrated and prepared for the study. The strands were washed and then bleached three times for 30 minutes each time using a solution of hydrogen peroxide (5.6% H₂O₂ + 13.9% (NH₄)₂S₂O₈, pH = 9.4). Healthy (unbleached) strands were kept as controls. After washing and air-drying at 55°C for 45 minutes, the strands were immersed in distilled water (as a control) or an aqueous solution containing 1% (w / v) of the hydrolyzed extract of milk thistle seed cake according to Example 1b for 24 h. Both solutions were pre-buried at pH 5.5. The strands were rinsed and then air-dried at approximately 60°C for 1 h.

[0307] According to the method described by FJ Wortmann and H. Deutz, Characterizing keratins using high-pressure differential scanning calorimetry (HPDSC), Vol. 48, No. 1, pp. 137-150, April 5, 1993, the denaturation temperature of human proteins was determined by differential calorimetry using a differential enthalpy analyzer (DSC Q100, TA Instruments) at a heating rate of 2 K / min.

[0308] The repair index relative to the placebo is calculated as follows:

[0309] (Bleached hair treated with the hydrolyzed extract according to the invention - control + water (placebo)) / (Control healthy hair - control + water (placebo)) × 100.

[0310] The repair index relative to the placebo is calculated as follows:

[0311] (Bleached hair treated with the hydrolyzed extract according to the invention - Control bleached hair) / (Control bleached hair - Control + Water (placebo)) × 100.

[0312] [Table 17]

[0313]

[0314] result:

[0315] Hair bleached with hydrogen peroxide is less stable than healthy, unbleached hair (Table 17). The difference in denaturation temperature of hair fiber proteins between damaged and healthy hair is -9.6°C.

[0316] Compared to water-treated damaged hair, treatment with 1% (w / v) of the extract of milk thistle seed cake according to Example 1b allowed for an improvement in hair protein stability against thermal denaturation by +5.4°C. The hydrolyzed extract of milk thistle seed cake according to Example 1b showed a repairing effect on hair fibers damaged by chemical oxidative stress compared to 64% of healthy hair.

[0317] in conclusion:Treatment of oxidized hair with the hydrolyzed extract of milk thistle seed cake according to the invention allows for improved thermal stability of hair fiber proteins. The hydrolyzed extract of milk thistle seed cake according to the invention exhibits a repairing effect on hair fibers damaged by hydrogen peroxide (oxidative stress), and thus is effective in repairing and / or maintaining and / or increasing the biomechanical properties of hair fibers, particularly in terms of strength, plasticity, resistance and / or flexibility.

[0318] The efficacy of 1% (w / v) of the hydrolyzed extract of milk thistle seed cake according to the invention in repairing cortex was 65% relative to the damaged control treated with water (placebo) and 69% relative to the damaged untreated control.

[0319] Example 11: In vitro effect of the extract according to the present invention on the strengthening of hair fibers (single fiber fatigue test)

[0320] Purpose: The effect of hydrolyzed extract of milk thistle seed cake according to Example 1a on the strengthening of brittle hair fibers was evaluated by fatigue testing of isolated hair fibers. Fatigue testing evaluates the tendency of hair to break under repeated applied force (Evans TA, Fatigue testing of hair—a statistical approach to hair breakage. J. Cosmet. Sci. 2009 Nov-December; 60(6):599-616). The mechanical tensile properties of hair reflect the internal state of its structure. This method can be considered a more realistic simulation of consumer practice, where combing represents this stimulus. This method also allows for the evaluation of the repairing effects of cosmetic ingredients, particularly on the repairing effects on the stratum corneum.

[0321] Materials and methods:

[0322] Strands of dark brown Caucasian hair bleached three times with hydrogen peroxide (1 g / 12 cm) were calibrated and cleaned for the study. Strands were treated with 1% (w / v) of milk thistle seed cake extract according to Example 1a mixed with distilled water for 1 hour at pH 5.0–5.5 or untreated, and then rinsed and dried. For each treated or untreated condition, fibers between 50 and 60 strands were studied under normalized conditions of 21°C ± 2°C and 40% ± 5% relative humidity.

[0323] In this test, individual hair fibers are subjected to repeated strain. The instrument counts the number of cycles of a predetermined stimulus necessary to induce breakage. A maximum number of cycles is defined. Under these test conditions, the fibers may or may not break during the experiment. The probability of survival at a given time is calculated using the Kaplan-Meier survival curve principle to estimate the number of surviving (or unbroken) keratin fibers.

[0324] Statistical analysis was performed to compare the groups using a nonparametric model. A p-value < 0.05 was considered significant.

[0325] The hair enhancement index is calculated using the following equation: (Number of cycles before breakage in treated bleached hair - Number of cycles before breakage in untreated bleached hair) / (Number of cycles before breakage in healthy hair - Number of cycles before breakage in untreated bleached hair) × 100

[0326] [Table 18]

[0327]

[0328] result:

[0329] The chemical oxidation process performed by hair bleaching significantly reduces the number of cycles that induce fiber breakage (Table 18). This indicates that bleached hair fibers are more fragile and prone to breakage.

[0330] Treatment of bleached hair three times with 1% (w / v) of the hydrolyzed extract of milk thistle seed cake according to Example 1a significantly increased the number of cycles prior to breakage, indicating enhancement and / or repair of hair damaged by chemical capillary oxidation.

[0331] in conclusion: The hydrolyzed extract of milk thistle seed cake according to the invention has properties that strengthen and thus repair hair fibers, making it possible to reduce the probability of breakage and thus maintain and / or increase the strength and / or plasticity and / or resistance and / or flexibility of hair fibers, especially repairing skin coverings, preferably hair.

[0332] Example 12: In vitro effects of the hydrolyzed extract of the present invention on the biomechanical properties of hair fibers – Yan Color retention (colorimetric method)

[0333] Purpose: The antioxidant and color-protecting effects of the hydrolyzed extract of milk thistle seed cake according to the present invention were evaluated in a hair color bleaching test after exposure to sunlight.

[0334] Materials and methods:

[0335] Hair strands were used from Caucasian hair that had been bleached twice with hydrogen peroxide and dyed with red pigment. The strands were calibrated (1 g / 15 cm), cleaned, and dried for the study. The strands were treated with a 1% (w / v) extract of milk thistle seed cake according to Example 1a mixed with distilled water for 1 hour or left untreated, 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. 2 The exposure lasted for 19 hours. The total exposure energy was 41,469 kJ / m² / cycle. After exposure, the hair strands were washed with water for 3 minutes and dried overnight in a chamber conditioned at 40% relative humidity and 21°C, followed by treatment according to L... a b The method involves colorimetric measurement. The parameter measured is ΔE. (Overall color change) and Δa related to red / green hues (Changes in red color). Measure the color of the hair strands after one, two, or three cycles of treatment and sun exposure. Use eight hair strands for each condition.

[0336] Statistical analysis of the data was performed by comparing each group with a Student's t-test. A p-value < 0.05 was considered significant.

[0337] Using stressed dyed and untreated groups as references, the percentage of efficacy in maintaining hair color against sun bleaching was obtained according to the following formula: 100 – [(dyed hair treated with the hydrolyzed extract according to the invention / control dyed hair) × 100]

[0338] [Table 19]

[0339]

[0340] result:

[0341] The overall color and redness of dyed hair exposed to solar radiation changed more and more during the exposure cycle (Tables 19 and 20). Sun exposure bleached the hair.

[0342] Treatment of dyed hair with a hydrolyzed extract of milk thistle seed cake according to Example 1a significantly slowed these color changes as a function of exposure cycles (Δa). and ΔE ).

[0343] The hydrolyzed extract of milk thistle seed cake according to the present invention has a protective effect on hair color against sun-induced bleaching and thus allows for the maintenance and / or enhancement of hair fiber color and / or shine.

[0344] Example 13: In vitro effects of the hydrolyzed extract of the present invention on the biomechanical properties of hair - anti-breakage

[0345] Purpose: The effectiveness of hair treatments in preventing breakage and split ends was tested on a device that allows for repeated combing of hair strands. The effects of chemical hair treatments such as bleaching on hair breakage can be examined. The protective effects of commercial reference products and the hydrolyzed extract according to the invention were then investigated.

[0346] Materials and methods:

[0347] A section of Caucasian brown hair (12 cm / 1 g) was calibrated, cleaned, and dried for the study. The section was bleached with 5% v / v hydrogen peroxide solution for 20 minutes, then rinsed and dried. The section was treated with a shampoo containing a hydrolyzed extract as a cosmetic ingredient according to Example 1c (0.25 g shampoo / g hair) and a conditioner containing the hydrolyzed extract as a cosmetic ingredient according to Example 1c (0.125 g conditioner / g hair). Prior to analysis, the products were rinsed and the section was allowed to stand at 40% relative humidity for 5 hours. The section was styled 50,000 times under the following conditions: 40% relative humidity, 30°C. Broken fibers less than 9 cm in length were collected and weighed.

[0348] Calculate the hair breakage value using the following formula: Broken hair (g) / Hair strand (g) × 100

[0349] Outliers were detected and removed from the data records using the Grubbs test at a significance level of 0.05.

[0350] Anti-breakage shampoo formula:

[0351] [Table 20]

[0352]

[0353] The formula for anti-breakage conditioner:

[0354] [Table 21]

[0355]

[0356] [Table 22]

[0357]

[0358] result:Compared to the 4.5% breakage percentage of brittle hair treated with shampoo and conditioner containing a hydrolyzed extract of milk thistle seed cake according to Example 1c, the breakage percentage of untreated brittle hair was estimated at 20.1% (Table 22). The finished product containing the hydrolyzed extract of milk thistle seed cake according to Example 1c had 78% protection against breakage.

[0359] in conclusion: Shampoos and conditioners containing a hydrolyzed extract of milk thistle seed cake according to the invention exhibit good conditioning properties and protect hair from breakage.

[0360] Example 14: In vitro effects of the hydrolyzed extract of the present invention on biomechanical properties – on the papillae of hair follicles Metabolic protection and stimulation

[0361] Papillary fibroblasts play a crucial role in coordinating 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-December;6(6):524-30. (1998)). With age, or in response to environmental stress, the physiology and metabolism of papillary fibroblasts may be affected, thus influencing the quality of the resulting keratin fibers.

[0362] Purpose:The aim of this study was to evaluate the anti-aging and metabolic-stimulating effects of a hydrolyzed extract of milk thistle seed cake according to Example 1a on a cultured model of papillary fibroblast aggregates derived from hair follicles by measuring ATP. This three-dimensional cell model allows the characteristics of the papillae to be preserved in vivo, particularly its ability to induce the formation and growth of high-quality hair (Yang CC and Cotsarelis G., Review of hair follicle dermal cells. J. Dermatol. Sci. 57(1): 2 (2010)).

[0363] Materials and methods:

[0364] A suspension consisting of fibroblasts (40 × 10³ cells) from human hair follicle papillae in DMEM containing 0.2% v / v growth factor (mesenchymal stem cell growth supplement) and a hydrolyzed milk thistle extract according to Example 1a, with or without 0.01% by weight / volume in distilled water, was placed in a 96-well microplate and centrifuged at 200 × g for 5 min to form aggregates. The aggregates were incubated at 37°C under a controlled atmosphere (5% CO2 and 95% relative humidity).

[0365] To analyze the parameters of interest, the aggregates were washed in buffered saline (PBS) and the cells were isolated by incubation in a mixture containing 100 mg of collagenase A and 20 mL of trypsin (2.5% v / v) and EDTA (0.02% v / v) in a 1:1 ratio.

[0366] According to the supplier's instructions (Bioluminescence Assay Kit CLS II Roche 11699695001, Sigma-Aldrich), use cells in suspension to measure ATP via bioluminescence.

[0367] result: Treatment of aggregates with a hydrolyzed extract of milk thistle seed cake according to Example 1a at 0.01% by weight / volume increased the total amount of ATP produced (+32% relative to the control; Table 23).

[0368] [Table 23]

[0369]

[0370] in conclusion:Treatment of papillary fibroblast aggregates with a hydrolyzed extract of milk thistle seed cake according to the invention allows for an increase in ATP within the pseudopapillary. These results indicate a potential for revitalizing keratin fibers in hair follicles, enhancing their structure and function.

[0371] Example 15: Shampoo and conditioner containing hydrolyzed extracts in cosmetic ingredient form according to Example 1c. Consumer testing of ingredients

[0372] Purpose: The aim of this study was to assess consumers’ perceptions of the benefits that hydrolyzed extracts, which are incorporated into shampoos and conditioners as cosmetic ingredients, provide to damaged hair.

[0373] Materials and methods:

[0374] The study was randomized under double-blind conditions and involved a total of 31 volunteers. The shampoos and conditioners used were the same as those used in Example 13 (Tables 20 and 21).

[0375] The same volunteers applied shampoo and conditioner or placebo products containing hydrolyzed extracts in the form of cosmetic ingredients at least three times a week for a period of four weeks, with a three-week treatment interruption between the two treatment phases.

[0376] Volunteers answered questionnaires about their perceptions of consumer benefits, measuring their opinions on the product after 4 weeks of use. A nonparametric binomial test was used to group and analyze the number of positive or negative questions (Chambers E. and Wolf M. "Sensory Testing Methods: Second Edition." Micro & Nano Letters (1996)). The significance threshold was set at 5% (p < 0.05).

[0377] result:

[0378] When volunteers regularly used shampoos and conditioners containing hydrolyzed extracts as cosmetic ingredients, the vast majority perceived beneficial effects on their hair. Among these positive reviews, the use of the hygiene and care program was considered to provide protection against chemical stress, breakage, and split ends. Hair appeared less damaged and healthier. Hair showed more vitality, volume, and shine, and was less dry. The scalp felt more comfortable.

[0379] In contrast, placebo shampoos and conditioners that do not contain hydrolyzed extracts as cosmetic ingredients do not show the vast majority of positive reviews associated with perceived benefits in protecting hair from chemical stress, split ends, increased vitality, increased volume, and reduced dryness.

[0380] [Table 24]

[0381]

[0382] According to the binomial test, the vast majority of positive reviews were received (p < 0.05).

[0383] in conclusion: Volunteers who regularly used shampoo and conditioner for 4 weeks reported additional benefits from the presence of hydrolyzed extracts, which are cosmetic ingredients, on their hair.

[0384] Example 16: Cosmetic Ingredients

[0385] Cosmetic ingredient 1 (by weight):

[0386] According to Example 1a), the hydrolyzed extract is 20%.

[0387] Maltodextrin 80%

[0388] Cosmetic ingredient 2 (by weight):

[0389] According to Example 1b), the hydrolyzed extract is 30%.

[0390] Maltodextrin 70%

[0391] Example 17: Cosmetic compositions containing cosmetic ingredients

[0392] Formula for anti-breakage shampoo

[0393] [Table 25]

[0394]

[0395] Formula for anti-breakage conditioner

[0396] [Table 26]

[0397]

Claims

1. Hydrolyzed extract of milk thistle seed cake is used for non-therapeutic cosmetic purposes to maintain and / or increase skin coverage, beneficial keratin fibers, and preferably the biomechanical properties of hair.

2. The use as described in claim 1, wherein, The hydrolyzed extract was obtained in water, which was the sole solvent.

3. The use as described in any of the preceding claims, wherein, The hydrolyzed extract was obtained through enzymatic hydrolysis.

4. The use as described in claim 3, wherein, The hydrolyzed extract was obtained by enzymatic hydrolysis at a pH between 3 and 10, preferably between 7 and 9.

5. The use as described in any of the preceding claims, wherein, The hydrolyzed extract has a total protein content of between 50% and 95% by weight, preferably between 60% and 90% by weight, and more preferably between 60% and 70% by weight, relative to the total dry weight of the hydrolyzed extract.

6. The use as described in any of the preceding claims, wherein, The hydrolyzed extract has a dry mass content of peptides with a weight average molecular weight of 100 Da to 20 kDa, which is between 50% and 90% by weight, advantageously between 65% and 90%, and more advantageously between 75% and 90% of the total dry mass of peptides in the hydrolyzed extract.

7. The use as described in any of the preceding claims, wherein, The hydrolyzed extract has a total dry mass content of peptides with a weight average molecular weight between 100 Da and 7000 Da, by weight, between 60% and 90%, advantageously between 70% and 80%, and more advantageously between 75% and 80%, of peptides with a weight average molecular weight between 100 Da and 700 Da relative to the total dry mass of the hydrolyzed extract.

8. The use as described in any of the preceding claims, wherein, The hydrolyzed extract maintains and / or increases the skin covering, preferably keratin fibers, advantageously the hair's resistance and / or strength and / or volume and / or color and / or shine and / or plasticity, i.e., non-brittle appearance, and / or smoothness.

9. The use as described in any of the preceding claims, wherein, This hydrolyzed extract is intended for topical application to the skin of the body and / or face, including skin coverings, particularly the scalp, and / or the nails, hair, body hair, especially beard, eyelashes and / or eyebrows, and more preferably all or part of keratin fibers, preferably all or part of hair.

10. The use as described in claim 9, wherein, Apply this product to damaged, dyed, or highlighted hair; dull, dry, brittle, fragile, brittle, fine, split, and / or stressed hair.

11. The use as described in any of the preceding claims, wherein, The hydrolyzed extract is atomized in the presence of maltodextrin at a weight concentration between 1% and 99%, advantageously between 5% and 90%, preferably between 40% and 80%, and more preferably between 70% and 80% relative to the total weight of the obtained powder.

12. The use as described in any of the preceding claims, wherein, The hydrolyzed extract, by weight, is 1 × 10⁻⁶ relative to the total weight of the cosmetic composition. -4 Between % and 10%, preferably 1 × 10 by weight -3 % to 5%, more preferably 1 × 10 by weight -3 % to 3%, or even more preferably 1 × 10 by weight -3 It is present in the cosmetic composition at a content of % to 1%, or advantageously 0.01% to 1% by weight.

13. The use as described in claim 12, wherein, The cosmetic composition comprises at least one cosmetically acceptable excipient, and wherein the composition is selected from serums, lotions, creams, shampoos, conditioners, oils, emulsions, ointments, pastes, foams, lotions, hydrogels, shower gels, face masks, hairsprays, sprays, waxes, mascaras, cosmetic brushes or nail polishes, advantageously shampoos, conditioners or lotions.

14. The use as claimed in claim 1, wherein the use is for repairing skin coverings, preferably hair.

15. A non-therapeutic cosmetic care method comprising topically applying a hydrolyzed extract of milk thistle seed cake or a cosmetic composition containing the same to maintain and / or increase skin coverage, beneficial keratin fibers, and preferably the biomechanical properties of hair.

16. The cosmetic care method of claim 14, wherein the method is used to repair the skin covering, advantageously the keratin fibers, and preferably the hair.

17. The beauty care method as described in claim 15 or 16, wherein, The hydrolyzed extract or the cosmetic composition comprising it is applied topically to the skin of the body and / or face, including all or part of the skin covering, particularly the scalp, and / or all or part of the skin covering, preferably all or part of the keratin fibers, more preferably all or part of the hair.

18. The beauty care method according to any one of claims 15 to 17, wherein, The extract is as defined in any one of claims 2 to 8 and 11 to 14.

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