Cosmetic use of hydrolyzed extract from milk thistle (Silybum marianum) seed cake

Abstract

The present invention relates to the cosmetic use of a hydrolyzed extract of milk thistle (Silybum marianum) seed cake to maintain and / or enhance the biomechanical properties of the skin's outer layer. Another subject of the present invention is a cosmetic care process comprising topical application of the hydrolyzed cake extract or a composition containing the same.

Description

[Technical Field]

[0001] This invention relates to the cosmetic use of a hydrolyzed extract of milk thistle (Silybum marianum) seed cake. [Background technology]

[0002] The care of the skin's outer layer, including nails, body hair, scalp hair, eyelashes, and eyebrows, involves comprehensive treatment, namely direct treatment of the corresponding skin areas, including not only keratin fibers but also the scalp and hair follicles (or hair bulbs and hair fibers).

[0003] Furthermore, direct care of hair fibers involves treating their constituent proteins. Hair proteins such as keratin are particularly sensitive to damage caused by lifestyle, exposure to chemical hygiene products such as their properties and frequency of use, styling habits, and environmental conditions such as sun, pollution, salt, wind, fluctuations in climate, general lifestyle, and age. These proteins denature, reducing the resistance of the outer layer of the skin, decreasing suppleness, making the hair brittle, and even dulling it, especially causing split ends. These layers then lose their biomechanical properties.

[0004] One solution is to provide exogenous hydrolyzable proteins to compensate for damage to keratin proteins and improve the moisture retention and elasticity of keratin fibers, especially hair fibers. Thus, the above proteins give the fibers greater flexibility, luster, suppleness, and body.

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

[0006] High-performance hydrolyzable proteins possess the property of adhering to hair fibers (weak bonding with 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 in the cosmetics market. These proteins are of plant or animal origin. The most common are hydrolyzed proteins derived from wheat, silk, keratin, collagen, elastin, milk, or almonds. However, there is a constant need for alternative high-performance ingredients in this field.

[0008] Quite unexpectedly, the applicant discovered that a hydrolyzed extract of milk thistle (Silybum marianum) seed cake has the ability to penetrate keratin fibers, thereby increasing their biomechanical properties. This hydrolyzed extract has been shown to act on the quality of the skin's outer layer, increasing its resistance, particularly reducing hair breakage, and making them more lustrous, more radiant, and larger in volume. Furthermore, the hydrolyzed extract of milk thistle (Silybum marianum) seed cake possesses skin-outer layer repair properties, as demonstrated in the following description.

[0009] An advantage of the hydrolyzed extract according to the present invention is that it is a co-product derived from industrial extraction residues, which are typically not upgraded, are considered industrial waste, and therefore fall within the scope of an environmentally responsible approach. Another advantage is that it is an ingredient that provides complete care, as it is effective on keratin fibers, particularly both hair and hair follicles. The hydrolyzed extract according to the present invention can also be readily manufactured on an industrial scale. Finally, enzymatic hydrolysis makes it possible to fractionate the proteins of the cake and recover low molecular weight peptides that can penetrate keratin fibers and provide the cosmetic effects described herein.

[0010] The hydrolyzed extract according to this invention is derived from the cake of the plant milk thistle (Silybum marianum). This plant, also known as milk thistle, has been known for thousands of years for its effects on the liver. It is still used in Asia and Africa for the treatment of kidney problems, gastrointestinal disorders, heart disease, rheumatism, and fever. The seeds are the primary part used for these therapeutic purposes.

[0011] The young stems, seeds, and fleshy buds are still conventionally consumed in some countries in the Middle East, North Africa, Sardinia, and Spain. The fruit of the milk thistle (Silybum marianum) can be used for oil extraction, as a dietary mixture with flour, in the preparation of various bakery products, or as a food.

[0012] Milk thistle (Silybum marianum) is also commonly used in cosmetics, particularly the flowers (without seeds) which are used to improve skin elasticity, firmness, barrier function, and moisture, as well as to reduce hair loss.

[0013] (Patent Document 1) discloses an extract of the fruit of the milk thistle used as a drug for hair growth and regeneration.

[0014] (Patent Document 2) discloses an extract of milk thistle (Silybum marianum) seeds obtained by heating the whole seeds and extracting with propylene glycol, which protects hair from damage caused by ultraviolet rays, particularly through its antioxidant properties that protect hair color and simultaneously improve resistance to hardness and tension.

[0015] Patent Document 3 describes a preparation process for obtaining an extract of Silybum marianum akene containing less than 0.2% silymarin. The method comprises extracting the oil with a solvent selected from an aqueous suspension, subcritical water, ethanol, isopropanol, or a mixture thereof.

[0016] Therefore, as far as the applicant knows, the prior art does not disclose the cosmetic use of the hydrolyzed extract according to the present invention. It is not suggested in any document to be used alone or in combination.

Prior Art Documents

Patent Documents

[0017]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0018]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

[0019] Therefore, the first subject relates to the non-therapeutic cosmetic use of hydrolyzed extract of milk thistle (Silybum marianum) seed cake to maintain and / or enhance the biomechanical properties of the skin, advantageously keratin fibers, and preferably hair.

[0020] The second subject concerns the non-therapeutic cosmetic use of the hydrolyzed extract according to the present invention in cosmetic compositions.

[0021] The third subject relates to a non-therapeutic cosmetic care process, which involves topical application of a hydrolyzed extract or cosmetic composition containing the present invention, for maintaining and / or increasing the biomechanical properties of the skin, preferably keratin fibers, and more preferably hair. [Means for solving the problem]

[0022] Therefore, the first subject relates to the non-therapeutic cosmetic use of hydrolyzed extract of milk thistle (Silybum marianum) seed cake to maintain and / or enhance the biomechanical properties of the skin, advantageously keratin fibers, and preferably hair.

[0023] The term "hydrolyzed extract" refers to the product obtained after enzymatic or chemical hydrolysis of milk thistle (Silybum marianum) seed cake in an aqueous solvent. In preferred embodiments, the hydrolyzed extract is obtained by enzymatic hydrolysis.

[0024] The term "cake" refers to the by-product of seed extraction, i.e., the residue obtained after oil extraction. Preferably, the milk thistle (Silybum marianum) seed cake used is a defatted milk thistle (Silybum marianum) seed cake.

[0025] The term "defatted" refers to a cake having an oil content of strictly less than 10% by weight, preferably strictly less than 8% by weight, relative to the total weight of the cake.

[0026] Generally, the term "oil" refers to all lipophilic compounds contained in the seeds of the milk thistle (Silybum marianum), including fat-soluble vitamins, fatty acids, mono-, di-, and triglycerides, as well as phospholipids.

[0027] The term "seed" refers to the fruit or akene that does not include its envelope. Therefore, this envelope, also known as the pericarp, is not included in the seeds according to this invention.

[0028] The term "cosmetic use" means non-therapeutic, non-medicinal, and non-dermatological use, i.e., use that does not require or include therapeutic treatment, and is intended for healthy skin, including the skin surface and / or skin surface.

[0029] The term “health” refers to the skin and / or skin that is classified as non-pathological by experts in this field, dermatologists, i.e., candidiasis, disease or condition, infection, inflammation, scarring, skin disease or condition, such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, especially seborrheic dermatitis, dandruff, or wounds or injuries and / or other skin diseases and / or alopecia and / or baldness and / or alopecia areata.

[0030] Therefore, for the purposes of the present invention, “damaged” skin is skin that is classified as “non-pathological” by those skilled in the art. Thus, they are both healthy and damaged. In particular, damaged skin is skin that has lost its flexibility and / or plasticity and / or resistance and / or strength and / or luster and / or color and / or volume, and as a result is particularly dehydrated. Thus, “damaged” skin has lost its biomechanical properties. Thus, they become dull, dry, less resistant, brittle, fragile, crackable and / or curved and / or branched. "Damage" to the skin surface may result from exposure to environmental conditions, such as wind, cold, salt, chlorine, pollution, sunlight, lifestyle, or even age, but it may also result from exposure to chemical or mechanical stress from daily treatments of the skin, such as rubbing, styling, curling, dyeing, bleaching, straightening, hair dryers, cosmetics, or sanitary chemicals, such as varnish, makeup and / or makeup removers, shampoos, conditioners, or creams.

[0031] Keratin fibers, especially hair, are also more difficult to style and shape. This loss of hair surface quality is visible and looks unattractive. Hair also reflects less light and is therefore noticeably less shiny, less colored, and less luminous. Hair is also finer and thinner. This is especially true for dyed or highlighted hair.

[0032] The term “skin outer layer” as used herein means head hair, eyelashes, eyebrows, body hair, especially beards, and / or nails. According to the present invention, beards include mustaches. Preferably, it refers to head hair.

[0033] The term "keratin fiber" refers to all fibers that make up the human hair system, particularly scalp hair, eyelashes, eyebrows, body hair, and especially facial hair including mustaches. Preferably, it refers to scalp hair.

[0034] The hydrolyzed extract according to the present invention is a locally acceptable component.

[0035] The term "topically acceptable" refers to an ingredient that is suitable for topical application, is non-toxic, does not irritate the skin, especially the scalp or skin surface, does not induce allergic or inflammatory reactions, and is not chemically unstable.

[0036] The hydrolyzed extract according to the present invention can be used topically.

[0037] The term "topical" means the direct topical application and / or vaporization of an ingredient to the skin, particularly the scalp, or the surface of the skin, particularly the hair.

[0038] Hydrolyzed extracts can be applied topically to all or part of the surface of the skin, particularly the scalp, or to all or part of the skin, more advantageously to the nails, hair, body hair, particularly the beard, eyelashes and / or eyebrows, more advantageously to the keratin fibers, and even more advantageously to the hair.

[0039] Accordingly, one subject of the present invention relates to the non-therapeutic cosmetic use of a hydrolyzed extract of milk thistle (Silybum marianum) seed cake for maintaining and / or increasing the biomechanical properties of the skin, preferably keratin fibers, and more preferably hair.

[0040] Within the scope of the present invention, the term "restoration of biomechanical properties" means at least partially improving and / or increasing the biomechanical properties of the skin exocutaneous tissue damaged by environmental, chemical, biological and / or mechanical stress, preferably by exposure to keratin fibers, and favorably to hair. In particular, the restored skin exocutaneous tissue can recover the biomechanical properties of the skin exocutaneous tissue before the damage occurred, i.e., the biomechanical properties of undamaged skin exocutaneous tissue. Therefore, it is possible to compare the effects of the hydrolyzed extract according to the present invention on these biomechanical properties, for example, by comparing the biomechanical properties of the skin exocutaneous tissue before and after treatment with the hydrolyzed extract according to the present invention, or by comparing the biomechanical properties of damaged skin exocutaneous tissue after treatment with the hydrolyzed extract according to the present invention with the biomechanical properties of undamaged skin exocutaneous tissue.

[0041] For the purposes of the present invention, the term "maintenance of biomechanical properties" means preventing and / or avoiding the deterioration of the biomechanical properties of the skin, preferably keratin fibers, and favorably hair, particularly when exposed to environmental, chemical, biological, and / or mechanical stress, by preventing the skin from becoming damaged skin and / or preventing these skins from being further damaged. Accordingly, for example, it is possible to compare the effect of the hydrolyzed extract according to the present invention on these biomechanical properties by comparing the biomechanical properties of skin treated with the hydrolyzed extract according to the present invention and exposed to environmental, chemical, biological, and / or mechanical stress with the biomechanical properties of skin not treated with the hydrolyzed extract according to the present invention and exposed to environmental, chemical, biological, and / or mechanical stress.

[0042] For the purposes of the present invention, the term "increase in biomechanical properties" means improving the biomechanical properties of the skin, preferably keratin fibers, favorably hair, in particular skin damaged by exposure to environmental, chemical, biological and / or mechanical stress, as well as undamaged skin. Therefore, it is possible to compare the effects of the hydrolyzed extract according to the present invention on these biomechanical properties by, for example, comparing the biomechanical properties of the skin before and after treatment with the hydrolyzed extract according to the present invention.

[0043] For the purposes of the present invention, the term “biomechanical properties” means the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e., their non-brittle appearance and / or flexibility, of the skin, preferably keratin fibers, and advantageously, hair.

[0044] Advantageously, the use of hydrolyzed extract of milk thistle (Silybum marianum) seed cake is beneficial for repairing the skin's outer layer, preferably keratin fibers, and advantageously, hair.

[0045] Preferably, the use of hydrolyzed extracts of milk thistle (Silybum marianum) seed cake is for maintaining and / or increasing the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e., non-brittleness and / or flexibility of the skin exoskeleton, preferably keratin fibers, and advantageously, hair.

[0046] These properties can be evaluated ex vivo by mechanical tests that measure the resistance of the skin, preferably keratin fibers, and more preferably hair, to tensile, stretching, bending, friction, and torsional forces. These biomechanical parameters, particularly those of hair, can be evaluated in response to stretching and can be measured, for example, by tensile tests (diastron). The parameters measured may be elastic modulus (Pa), elongation at break (%), breaking force (gmf), and gradient of the plastic strain region (gmf / % elongation), normalized with respect to the diameter of the skin. This last parameter (gradient) allows for the measurement of the plasticity of the study material immediately before failure. This technique provides a stress (or force) curve as a function of the characteristic elongation of the material under test. It is known that an increase in elongation at break and a decrease in the plastic strain gradient can be observed on hair fibers (skin) damaged by oxidation, reduction, or UV irradiation after cleaving disulfide crosslinks to free radicals in the cortical domain.

[0047] In one embodiment of the present invention, "maintaining and / or increasing the biomechanical properties" of the skin's outer layer means reducing advanced glycation end products (AGEs), which are prominently present during time-induced or photo-induced aging upon contact with chemical or physical oxidizing agents, but also appear daily depending on environmental conditions. These glycation products degrade the biomechanical properties of the skin's outer layer.

[0048] Environmental stresses include smoke, especially cigarette smoke; pollution, especially metals, PM2.5 and PM10 particulate matter; temperature, especially heat and cold and rapid temperature changes; moisture, especially humidity or dryness; solar radiation, especially the visible spectrum, UV and / or gamma rays and / or blue light; rain, wind, dust, and sea salt; swimming pool water, especially chlorine and transition metals present in the water; but also include intrinsic and / or time-induced aging. The latter is also involved in the loss of structural and / or functional quality of the skin crust, preferably keratin fibers, especially eyelashes, eyebrows and scalp hair, more specifically scalp hair.

[0049] Chemical stress includes active convenience, hygiene and beauty products, especially active shampoos and hair care and / or hair treatment products, particularly those for styling and shaping, such as straightening and / or permanent waves and / or dyeing and / or bleaching, chlorine and transition metals present in swimming pool water, varnishes and / or solubilizers, solvents or other chemical agents such as makeup products.

[0050] Mechanical stresses include, in particular, friction such as brushing and / or rubbing against fabrics (pillows or clothing) and / or particles such as dust and / or sand, heat from hair dryers and straightening irons, and / or styling, and especially exposure to tensile, stretching and / or torsional forces.

[0051] Accordingly, in one embodiment of the present invention, the ability of the extract according to the present invention to “repair” damaged skin crust can be evaluated according to existing art. Conventional methods for measuring restorative effects allow for the measurement of the ability of a product to be evaluated to restore the visual, structural, and / or functional state of damaged skin crust, preferably damaged hair, which is comparable to the state of undamaged skin crust, preferably undamaged hair. These are performed on damaged (or impaired) skin crust, preferably hair, and the restorative effect is measured by comparison with undamaged (or impaired) skin crust, preferably hair. Advantageously, the restorative effect is the restorative effect on skin crust damaged (or impaired) in vitro with an oxidizing agent that induces denaturation of crustal proteins. Advantageously, this is the restorative effect of the hydrolyzed extract according to the present invention on hair, more preferably hydrogen peroxide as the oxidizing agent. More advantageously, this protein denaturation is evaluated by measuring the protein denaturation temperature (°C) by differential calorimetry using a differential scanning calorimeter (DSC Q100, TA Instruments) under the conditions described in Example 10 (Non-Patent Literature 1).

[0052] In vivo, advantageously in hair, several methods can be used to evaluate this restorative effect on the skin epithelium, selected from video microscopy, confocal microscopy, FTIR or Raman microscopy, X-ray, and electron microscopy, which are used in particular to observe the condition and / or quality of the cuticle and the protective envelope of keratin fibers in the skin epithelium. Microscopic studies of the whole hair allow for the visualization and quantification of detached scale as evidence of surface damage. These methods can also be used on hair cross-sections to observe the condition and / or quality of the keratin fiber cortex. Physical surface quantification methods can be performed to evaluate the morphology of skin appendages, especially hair, such as atomic force microscopy or white light interference geometry, to assess chemistry (XPS), charge (flow potential), or energy (reverse gas chromatography).

[0053] Calorimetry (DSC) can be used to evaluate the restorative effect of the hydrolyzed extract according to the present invention on the internal properties of the skin crust, particularly hair.

[0054] Within the scope of the present invention, the term "maintain and / or increase volume" means maintaining and / or increasing the thickness of the skin epidermis over its entire length, preferentially maintaining and / or increasing the diameter of keratin fibers, and preferably the diameter of hair. [Modes for carrying out the invention]

[0055] The term “maintaining and / or increasing the biomechanical properties” of the skin exoskeleton means, for the purposes of the present invention, 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 nonbrittleness and / or their flexibility. These properties are maintained and / or increased by the following methods described in the following paragraphs and corresponding examples. - Particularly by measuring free radical activity in vitro, as described in Example 3; - Particularly by measuring the chelating activity to transition metals such as iron (Fe) in vitro, as described in Example 4; - Particularly by measuring the chelating activity to transition metals such as copper (Cu) in vitro, as described in Example 5; - Particularly by measuring anti-glycation activity against cutaneous proteins in vitro, as described in Example 6; - Particularly by measuring the carbonylation of keratin and related skin epidermal proteins ex vivo, as described in Examples 7a and b); -By measuring the hair protection score (HPS) with ExVivo, as described in Example 7d; -By measuring the stabilization of the cysteine ​​group ex vivo, as in Example 9; -By exvivo, as in Examples 11 and 13, by repeated hair styling tests that allow evaluation of tensile strength tests (Diastron), fatigue tests, or their failure; -By measuring the color change of the skin's outer layer ex vivo, as in Example 12; -By measuring the amount of ATP in fibroblasts in the papilla of the hair follicle in vitro, as in Example 14; It can be measured.

[0056] 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 present invention. Therefore, in a particularly preferred embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if, under the conditions described in Example 3, the oxidation inhibition rate of free radical DPPH° in the presence of the hydrolyzed extract according to the present invention is greater than 50%, preferably greater than 65%, more preferably greater than 80%, and more preferably greater than 90%, compared to the oxidation inhibition rate of free radical DPPH° measured in the absence of the hydrolyzed extract according to the present invention. Preferably, the skin crust is keratin fiber, and preferably hair.

[0057] These properties can also be evaluated in vitro by measuring chelating activity to transition metals such as iron (Fe) by, for example, a tryptophan degradation test and / or dityrosine and / or pentosidine and / or fluorescent glycation product formation test, measured in the presence of hydrogen peroxide and iron as oxidation catalysts, after oxidation of albumin used as a protein model, in the presence of the hydrolyzed extract according to the present invention.

[0058] In alternative embodiments, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if the protective index calculated for the tryptophan degradation test under the conditions of Example 4 is greater than 10%, preferably greater than 15%, more preferably greater than 20%, particularly greater than 25%, more particularly greater than 30%, and even more particularly greater than 35%. Preferably, the skin crust consists of keratin fibers, preferably hair.

[0059] In another embodiment, if, in the presence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 4, the rate of dityrosine formation is reduced by at least 10%, preferably at least 15%, more preferably at least 20%, preferably at least 25%, compared to the rate of dityrosine formation measured in the absence of the hydrolyzed extract according to the present invention, and / or if the protection index calculated for the dityrosine formation test under the conditions of Example 4 is greater than 30%, preferably greater than 35%, more preferably greater than 60%, particularly greater than 65%, then the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust. Preferably, the skin crust is keratin fibers, preferably hair.

[0060] In another embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if the rate of pentosidine formation in the presence of the hydrolyzed extract according to the present invention is reduced by at least 10%, preferably at least 25%, compared to the rate of pentosidine formation measured in the absence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 4, and / or the protective index calculated for the pentosidine formation test under the conditions of Example 4 is greater than 50%, preferably greater than 60%. Preferably, the skin crust is keratin fiber, preferably hair.

[0061] In another embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if, in the presence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 4, the amount is at least 5%, preferably at least 10%, more preferably at least 13%, compared to the rate of formation of fluorescent glycation products measured in the absence of the hydrolyzed extract according to the present invention, and / or the protection index calculated under the conditions of Example 4 for the fluorescent glycation product formation test is greater than 10%, preferably greater than 15%, more preferably greater than 20%, and particularly greater than 25%. Preferably, the skin crust is keratin fibers, preferably hair.

[0062] The biomechanical properties of the present invention can also be evaluated in vitro by measuring the chelating activity to transition metals such as copper (Cu) in a lipid oxidation test, for example, by measuring the formation of malondialdehyde (MDA) in the presence of copper (Cu) (oxidation catalyst) and thiobarbituric acid (TBA) in the presence of the hydrolyzed extract of the present invention. In another embodiment, the hydrolyzed extract of the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if the proportion of oxidized lipids by assaying MDA in the presence of the hydrolyzed extract of the present invention, preferably under the conditions described in Example 5, is reduced by at least 20%, favorably at least 40%, more favorably at least 60%, preferably at least 75%, compared to the proportion of lipids oxidized by an MDA assay measured in the absence of the hydrolyzed extract of the present invention, and / or if the protection index calculated under the conditions of Example 5 for a fluorescent glycation product formation test is greater than 60%, favorably greater than 75%, more favorably greater than 90%, particularly greater than 95%. Favouritely, the skin crust is keratin fiber, preferably hair.

[0063] These properties can also be evaluated in vitro, for example, by measuring the anti-glycation activity on skin crust proteins using a dityrosine and / or pentosidine formation test measured after incubation of the crust in a glucose solution in the presence of the hydrolyzed extract according to the present invention. In another embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if, in the presence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 6, the rate of dityrosine formation is reduced by at least 5%, preferably at least 10%, and more preferably at least 15%, compared to the rate of dityrosine formation measured in the absence of the hydrolyzed extract according to the present invention, and / or if the protection index calculated for the dityrosine formation test under the conditions of Example 6 is greater than 40%, preferably greater than 45%, more preferably greater than 50%, and especially greater than 55%. Preferably, the skin crust is keratin fiber, preferably hair.

[0064] In another embodiment, if, in the presence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 6, the pentosidine formation rate decreases by at least 5%, preferably at least 10%, and more preferably at least 15%, compared to the pentosidine formation rate measured in the absence of the hydrolyzed extract according to the present invention, and / or if the protection index calculated for the pentosidine formation test under the conditions of Example 6 is greater than 40%, preferably greater than 45%, more preferably greater than 50%, and particularly greater than 55%, then the hydrolyzed extract according to the present invention is in an effective amount for "maintaining and / or increasing the biomechanical properties" of the skin crust. Preferably, the skin crust is keratin fibers, preferably hair.

[0065] Biomechanical properties can also be evaluated ex vivo by measuring the carbonylation of keratin and its related proteins, for example by particle (contamination) and UV tests measured in the presence of the hydrolyzed extract according to the present invention. In another embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the biomechanical properties" of the skin crust if, in the presence of the hydrolyzed extract according to the present invention, particularly in the cuticle, and more specifically in the cortex, and preferably under the conditions described in Example 7, the percentage of carbonylated proteins measured in the absence of the hydrolyzed extract according to the present invention is reduced by at least 5%, preferably at least 15%, more preferably at least 20%, more preferably at least 25%, and more preferably at least 30%, and / or if the protection index calculated under the conditions of Example 7 is greater than 40%, preferably greater than 45%, more preferably greater than 50%, and especially greater than 55%. Preferably, the skin crust is keratin fibers, preferably hair.

[0066] Biomechanical properties can also be evaluated in vitro by measuring oxidative stress in the skin, for example, by optical measurement of birefringence properties, particularly the birefringence (Kindex) of the skin skin treated with the hydrolyzed extract according to the present invention. In another embodiment, the hydrolyzed extract according to the present invention is an effective amount for "maintaining and / or increasing the biomechanical properties" of the skin skin if, in the presence of the hydrolyzed extract according to the present invention, the Kindex value under the conditions preferably described in Example 7c increases by at least 5%, preferably at least 10%, more preferably at least 13%, compared to the Kindex value measured in the absence of the hydrolyzed extract according to the present invention, and / or if the improvement index calculated under the conditions of Example 7c is greater than 40%, preferably greater than 45%, more preferably greater than 50%, particularly greater than 55%. Preferably, the skin skin is keratin fiber, preferably hair. More preferably, it is the hydrolyzed extract prepared according to Example 1a).

[0067] Biomechanical properties can also be evaluated exvivo by measuring the Hair Protection Score (HPS) and classifying hair care products on damaged skin exodermis (UVA irradiation and particles) treated with the hydrolyzed extract according to the present invention. In another embodiment, the hydrolyzed extract according to the present invention is an effective amount for "maintaining and / or increasing the biomechanical properties" of the skin exodermis when the HPS is greater than 30, preferably 50 or more, preferably under the conditions described in Example 7d. Advantageously, the skin exodermis consists of keratin fibers, preferably hair.

[0068] These properties can be further evaluated ex vivo by measuring the stabilization of cysteine ​​groups, for example, by measuring the vibrational spectrum of damaged skin epidermal keratin treated with the hydrolytic extract according to the present invention. In another embodiment, the hydrolytic extract according to the present invention is used to measure the vibrational spectrum of damaged skin epidermis at 1174, 1116, 1150 and / or 840 cm⁻¹. -1The absorption bands were 1174, 1116, 1150 and / or 840 cm² measured in the presence of the hydrolyzed extract according to the present invention, preferably under the conditions described in Example 9, on the decolorized skin surface in the absence of the hydrolyzed extract according to the present invention. -1 An amount effective for "maintaining and / or increasing the biomechanical properties" of the skin crust is when the absorption band is reduced by at least 5%, preferably at least 15%, more preferably at least 20%, and even more preferably at least 35%, and / or when the effectiveness index calculated under the conditions of Example 9 is greater than 5%, preferably at least 10%, and more preferably at least 13%. Preferably, the skin crust consists of keratin fibers, preferably hair.

[0069] Biomechanical properties, particularly strength and / or plasticity and / or resistance and / or flexibility, can also be evaluated ex vivo by measuring the stability of the damaged skin exocutaneous, for example by measuring the denaturation temperature of proteins in the damaged skin exocutaneous, using hydrogen peroxide, and then treating with the hydrolytic extract according to the present invention. In another embodiment, the hydrolytic extract according to the present invention is an effective amount for "maintaining and / or increasing the strength and / or flexibility and / or resistance and / or plasticity" of the skin exocutaneous if, in the presence of the hydrolytic extract according to the present invention, preferably under the conditions described in Example 10, the denaturation temperature is increased by at least 2°C, preferably at least 5°C, compared to the denaturation temperature measured in the absence of the hydrolytic extract according to the present invention, and / or the repair index for placebo-treated or untreated skin exocutaneous calculated under the conditions of Example 10 is greater than 40%, preferably greater than 45%, more preferably greater than 50%, particularly greater than 60%. Favouritely, the skin exocutaneous is keratin fiber, preferably hair.

[0070] These properties can also be evaluated by tensile strength tests (Diastron), fatigue tests, or repeated hair styling tests that allow for the assessment of their breakage. It is also possible to quantify hair fibers with split ends. Furthermore, these biomechanical properties can be evaluated by measuring the styling force required for dry or wet hair. In particular, these properties can be evaluated ex vivo by measuring the rupture of the skin exoskeleton by fatigue tests on damaged skin exoskeleton, for example, preferably treated with hydrogen peroxide and then with the hydrolyzed extract according to the present invention. In another embodiment, the hydrolyzed extract according to the present invention is an effective amount for "maintaining and / or increasing the strength and / or plasticity and / or resistance and / or flexibility" of the skin exoskeleton if, under the conditions described in Example 11, the number of cycles before skin exoskeleton breakage or the hair fiber strengthening index in the preferential presence of the hydrolyzed extract according to the present invention is greater than 10%, preferably greater than 20%, and more preferably greater than 30% compared to the number of cycles before skin exoskeleton breakage or the strengthening index measured in the absence of treatment with the hydrolyzed extract according to the present invention. Advantageously, the skin exocutaneous tissue consists of keratin fibers, preferably hair. More advantageously, it is a hydrolyzed extract prepared according to Example 1a).

[0071] In another embodiment, the hydrolyzed extract according to the present invention is effective in "maintaining and / or increasing the strength and / or flexibility and / or resistance and / or flexibility" of the skin if, after repeated styling of damaged skin exodermis and in the presence of the hydrolyzed extract according to the present invention, the reduction in the rate of breakage, preferably under the conditions described in Example 13, is greater than 50%, preferably greater than 70%, and more preferably greater than 75%, compared to the rate of breakage of the skin exodermis measured in the absence of treatment with the hydrolyzed extract according to the present invention. Advantageously, the skin exodermis is keratin fiber, preferably hair. More advantageously, the hydrolyzed extract is prepared according to Example 1c).

[0072] The biomechanical properties of color and / or gloss can be evaluated ex vivo, for example, by measuring the change in color of damaged skin exoskeleton by a colorimetric test (delta a* and delta E*), preferably using hydrogen peroxide, after exposure to sunlight, and then treating with the hydrolytic extract according to the present invention. In another embodiment, the hydrolytic extract according to the present invention is effective in "maintaining and / or increasing the color and / or gloss" of skin exoskeleton if, in the presence of the hydrolytic extract according to the present invention, the delta a* value is increased by 4%, preferably 6%, compared to the delta a* value measured in the absence of the hydrolytic extract according to the present invention, and / or the effectiveness % calculated for delta a* under the conditions of Example 12 is at least 20%, favorably at least 30%, particularly at least 80%, and / or the delta E* value is decreased by 2%, preferably 3%, compared to the delta E* value measured in the absence of treatment with the hydrolytic extract according to the present invention, and / or the effectiveness % calculated for delta E* under the conditions of Example 12 is at least 15%, favorably at least 20%, particularly at least 25%. Advantageously, the outer layer of skin is made up of keratin fibers, with hair being the preferred material.

[0073] Biomechanical properties can also be evaluated in vitro by measuring the amount of ATP in the fibroblasts of the hair follicle papillae. In another embodiment, the hydrolyzed extract according to the present invention is an effective amount for "maintaining and / or increasing the biomechanical properties" of the skin crust, preferably keratin fibers, and preferably hair, if, in the presence of the hydrolyzed extract according to the present invention, preferably in the presence of the hydrolyzed extract prepared according to Example 1a, under the conditions described in Example 14, the proportion of fibroblast aggregates in the papillae is greater than 10%, preferably greater than 20%, more preferably greater than 25%, and favorably greater than 30%, compared to the proportion of fibroblast aggregates in the papillae measured in the absence of treatment with the hydrolyzed extract according to the present invention.

[0074] Therefore, the hydrolyzed extract according to the present 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 gloss and / or plasticity of the skin, preferably hair, thereby reducing their fragility, breakage and roughness, and thus improving tangling and / or styling.

[0075] The term “maintenance and / or enhancement of biomechanical properties” of the skin, and more favorably, of hair, means making the skin, and more favorably, of hair visually smoother and / or shinier and / or brighter and / or easier to manage and / or less prone to split ends. These properties may be evaluated by external volunteers trained to define visual or tactile sensations, by imaging techniques such as video or electron microscopy, or by sensory testing of hair strands, or by evaluation tests conducted by professionals (e.g., beauticians) or trained volunteers, or by self-assessment using consumer questionnaires. The efficacy of the product in the formulation is evaluated according to perceptible visual or tactile quality criteria, particularly the head of the hair.

[0076] The increased restorative and / or biomechanical properties of the hydrolyzed extract according to the present invention make it a cosmetic active ingredient for improving the resistance and / or strength and / or volume and / or color and / or gloss and / or luster of the skin crust, particularly keratin fibers, and preferably hair.

[0077] The enzymatic hydrolysis for obtaining the hydrolyzed extract according to the present invention may be carried out in the presence of any protease known to those skilled in the art, preferably an enzyme of pancreatic origin such as pepsin, trypsin, or chymotrypsin, preferably an enzyme of animal origin selected from plant-derived enzymes preferably selected from trypsin, papain, bromelain, ficin, actinidine, preferably papain, or an enzyme from the Bacillus licheniformis strain sold under the name Alcalase®, or a bacterial enzyme selected from the B. subtilis strain, preferably an enzyme from the Bacillus licheniformis strain sold under the name Alcalase®. In a particularly advantageous embodiment of the present invention, the enzyme used is derived from the Bacillus licheniformis strain and is particularly sold under the name Alcalase®.

[0078] The amount of Silybum marianum seed cake used in hydrolysis according to the present invention is preferably 1% to 20%, more preferably 2% to 15%, and even more preferably 10% to 15%, relative to the total weight of the solvent and cake.

[0079] The solvent that can be used for hydrolysis may be an aqueous solvent.

[0080] According to the present invention, the term "aqueous solvent" means an aqueous extract solution containing more than 60% by weight, preferably at least 70% by weight, particularly at least 80% by weight, more specifically at least 90% by weight, particularly at least 95% by weight of water, based on the total weight of the aqueous solution, and more preferably free of glycol and particularly free of alcohol. The aqueous solvent does not contain a suspension aqueous solution, subcritical water, ethanol, isopropanol, or propylene glycol.

[0081] Advantageously, the only solvent used is water.

[0082] Hydrolysis can be carried out at a pH of 3 to 10, preferably 7 to 9, very preferably 7.5 to 8.5, and even more preferably 7.5, depending on the optimal pH of the enzyme.

[0083] Alternatively, enzymatic hydrolysis can be carried out in the presence of papain at a pH of 3-5.5, preferably 4.5-5.

[0084] Hydrolysis can be carried out at temperatures of 30°C to 75°C, preferably 50°C to 70°C, especially 55°C to 65°C, and most preferably 65°C. This can be carried out over a period of 30 to 24 hours, preferably 45 minutes to 12 hours, more preferably 50 minutes to 6 hours, and very preferably 1 hour to 3 hours.

[0085] The amount of enzyme used for hydrolysis is 0.1% to 25% by weight relative to the weight of the cake, preferably 0.5% to 20% by weight, more preferably 0.75% to 10% by weight, and favorably 1% to 5% by weight.

[0086] Next, the enzyme to be used can be inactivated by heating, preferably at a temperature of 80°C to 100°C, and very preferably at 90°C, for a period of 10 minutes to 3 hours, more preferably 10 minutes to 2 hours, and more preferably 20 minutes to 1 hour 30 minutes. Enzyme inactivation can occur at a pH of 3.5 to 8, preferably 4 to 7, more preferably 4 to 6.5, and preferably 6.0.

[0087] Next, the obtained hydrolyzed extract can be centrifuged and then clarified by continuous filtration to a porosity (filtration threshold) of 0.22 μm.

[0088] Therefore, the hydrolyzed extract according to the present invention may be in liquid form containing a dry matter content of advantageously 1% to 20% by weight, advantageously 2% to 10% by weight, and more advantageously 3% to 6% by weight, relative to the total weight of the hydrolyzed extract in liquid form.

[0089] In another embodiment, the aqueous extraction step is carried out prior to the hydrolysis step by any method known to those skilled in the art, particularly by maceration, and preferably by extraction using water as the sole solvent. The amount of cake used for the aqueous extraction is 1% to 20%, preferably 2% to 15%, and more preferably 10% to 15%, relative to the total weight of the solvent and cake.

[0090] In this case, aqueous extraction may be performed at a pH of 7.5–12, preferably at a pH of 8–9, very preferably at a pH of 9, for a period of 30 minutes to 2 hours, and preferably for 1 hour.

[0091] Aqueous extraction can be carried out at temperatures of 4°C to 300°C, preferably 15°C to 100°C, more preferably 20°C to 90°C, and very preferably 20°C to 70°C. In a particularly advantageous embodiment of the present invention, aqueous extraction is carried out at room temperature, i.e., 20 to 25°C.

[0092] In a preferred embodiment, aqueous extraction is carried out at 20°C and pH 12 using an amount of water equal to 15% of the weight of the cake relative to the total weight of the cake and water.

[0093] In another embodiment, aqueous extraction is performed at 20°C and pH 12 using an amount of 12.5% ​​of the cake's weight relative to the total weight of the cake and water.

[0094] In another embodiment, aqueous extraction is performed under subcritical conditions.

[0095] The term "extraction under subcritical conditions" refers to extraction in the presence of water under temperature conditions above 100°C and pressure conditions below 22.1 MPa (221 bar). The water remains in a liquid state but has lower viscosity and surface tension than water at room temperature, and its dielectric constant is increased. Therefore, the extraction pressure is 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).

[0096] Therefore, under subcritical conditions, extraction is carried out in water, particularly at temperatures in the range of 100°C to 300°C, preferably 120°C to 250°C, and more preferably 120°C to 180°C. Extraction can be carried out at a single given temperature or at continuously increasing temperatures. In an advantageous embodiment of the present invention, extraction is carried out at a single temperature of 160°C. In another embodiment, this is carried out according to a gradient of three increasing temperatures, for example, 100°C to 200°C, 120°C, 140°C, then 160°C, or 110°C, 130°C, then 150°C, or otherwise 120°C, 145°C, then 170°C.

[0097] In a preferred embodiment, subcritical extraction is performed at a pressure of 5 MPa (50 bar) and a temperature of 120°C for 20 minutes. The resulting extract is then filtered while still hot.

[0098] In another embodiment, subcritical extraction is performed at a pressure of 5 MPa (50 bar) and a temperature of 140°C for 20 minutes. The resulting extract is then filtered while still hot.

[0099] In another embodiment, the hydrolyzed extract of the milk thistle (Silybum marianum) seed cake obtained in this way can be used as a solvent for hydrolyzing a new amount of milk thistle (Silybum marianum) seed cake.

[0100] In a favorable embodiment, the hydrolyzed extract is obtained by enzymatic digestion as follows: Milk thistle (Silybum marianum) seed cake is crushed and suspended in water (as the sole solvent) at a content of 12.5% ​​by weight of the seed cake relative to the total weight of the cake and solvent, and subjected to hydrolysis at a temperature of 55°C and pH 8.5 for 1 hour, maintaining the pH during hydrolysis, and the weight concentration of bacterial protease (Alcalase®) derived from the liquid form of Bacillus licheniformis strain relative to the weight of the cake is 1%. The enzyme is then inactivated by heating at a temperature of 90°C and pH 4.5 for 1 hour. The mixture is then cooled, centrifuged, and the supernatant is filtered under the conditions described in Example 1b) (filtration threshold: 0.22 μm).

[0101] In a particularly advantageous embodiment of the present invention, the hydrolyzed extract is obtained by enzymatic digestion as follows: Milk thistle (Silybum marianum) seed cake is crushed and suspended in water (as the sole solvent) at a content of 15% by weight of the seed cake relative to the total weight of the cake and solvent, and subjected to hydrolysis at a temperature of 65°C and pH 7.5 for 2 hours, maintaining the pH during hydrolysis, and the weight concentration of bacterial protease (Alcalase®) derived from the liquid form of Bacillus licheniformis strain relative to the weight of the cake is 1.7%. The enzyme is then inactivated by heating at a temperature of 90°C and pH 6.0 for 1 hour. The mixture is cooled to room temperature (20°C-30°C), centrifuged, and filtered under the conditions described in Example 1a) (filtration threshold: 0.22 μm).

[0102] The term "small peptide" or "low molecular weight peptide" refers to peptides with a weight-average molecular weight of less than 7000 daltons, analyzed by gel permeation chromatography (e.g., Superdex® peptide 10 / 30HR GE or Superose® 12 10 / 300GL), followed by column calibration using protein molecules of known molecular weight (gel permeation chromatography (GPC)).

[0103] Advantageously, the hydrolyzed extract according to the present invention has a total protein content of 50% to 95% by weight, preferably 60% to 90% by weight, and more preferably 60% to 70% by weight, based on the total dry mass of the hydrolyzed extract according to the present invention.

[0104] Advantageously, the hydrolyzed extract according to the present invention has a dry weight content of peptides having a weight-average molecular weight of 100 Da to 20 kDa, as analyzed by gel permeation chromatography (Superose® 12 10 / 300 GL, etc.), which is 50% to 90% by weight, preferably 65% ​​to 90% by weight, and more preferably 75% to 90% by weight, relative to the total dry weight of the hydrolyzed peptides in the hydrolyzed extract according to the present invention.

[0105] Advantageously, the hydrolyzed extract according to the present invention has a dry weight content of peptides having a weight-average molecular weight of 100 Da to 700 Da, as analyzed by gel permeation with a Superdex® peptide 10 / 30 HR GE column, which is 60% to 90% by weight, advantageously 70% to 80% by weight, and more advantageously 75% to 80% by weight, relative to the total dry weight of peptides having a molecular weight of 100 Da to 7000 Da in the hydrolyzed extract according to the present invention (column sensitivity).

[0106] Advantageously, the hydrolyzed extract according to the present invention has a dry weight content of peptides having a weight-average molecular weight of 100 Da to 2500 Da, as analyzed by gel permeation with a Superdex® peptide 10 / 30HR GE column, which is 70% to 99% by weight, advantageously 80% to 99% by weight, and more advantageously 90% to 99% by weight, relative to the total dry weight of peptides having a molecular weight of 100 Da to 7000 Da in the hydrolyzed extract according to the present invention (column sensitivity).

[0107] Even more advantageously, the hydrolyzed extract according to the present invention is not used in combination with retinoids.

[0108] The hydrolyzed extract according to the present invention may be used alone or incorporated into a cosmetic composition.

[0109] In one embodiment, the hydrolyzed extract can then be dried, for example, by freeze-drying or spraying, in or out of the presence of maltodextrin. The hydrolyzed extract is then in powder form.

[0110] According to one embodiment, the hydrolyzed extract according to the present invention obtained under the conditions described in Examples 1a) and 1b) in particular can be atomized at a weight concentration of maltodextrin of 1% to 99%, preferably 5% to 90%, preferably 40% to 85%, more preferably 65% ​​to 85%, and more preferably 70% to 80%, relative to the total weight of the obtained powder, and preferably under the conditions of Example 1c) or 1d).

[0111] Alternatively, the hydrolyzed extract may be used alone in solid form in the form of a cosmetic or dermatological ingredient.

[0112] Alternatively, when used alone in liquid form as a cosmetic or dermatological ingredient, it is dissolved in an aqueous solution containing glycerol, which is favorably present at a concentration of 60% to 90% by weight, more favorably at a concentration of 70% to 85% by weight, and very favorably at a concentration of 80% by weight, relative to the total weight of the aqueous solution containing the hydrolyzed extract.

[0113] In alternative embodiments of the present invention, the hydrolyzed extract is dissolved and / or diluted in a solvent, particularly a polar solvent, such as water, alcohol, polyol, glycol, such as pentylene glycol and / or butylene glycol and / or hexylene glycol and / or caprylyl glycol, or a mixture thereof, preferably in a water-glycol mixture, and more preferably containing a glycol selected from hexylene glycol, propylene glycol, caprylyl glycol, and any mixture thereof. Advantageously, the hydrolyzed extract according to the present invention is dilutable and / or soluble in an aqueous solution containing hexylene glycol, particularly 0.1% to 10% by weight of hexylene glycol, preferably 0.5% to 5% by weight of hexylene glycol, relative to the total weight of the cosmetic ingredients. Advantageously, the hydrolyzed extract according to the present invention is dilutable and / or soluble in an aqueous solution containing caprylyl glycol, particularly 0.01% to 5% by weight, preferably 0.1% to 1% by weight of caprylyl glycol, relative to the total weight of the aqueous solution containing the hydrolyzed extract. Alternatively, the solution in which the hydrolyzed extract according to the present invention is dissolved contains pentylene glycol and caprylyl glycol.

[0114] In particular, the aqueous solution in which the hydrolyzed extract according to the present invention is dissolved contains xanthan gum, especially 0.01% to 5% by weight of xanthan gum relative to the total weight of the aqueous solution, and more specifically 0.1% to 1% by weight of xanthan gum relative to the total weight of the aqueous solution containing the hydrolyzed extract.

[0115] Hydrolyzed extracts can be used alone in the form of cosmetic ingredients, or incorporated into cosmetic compositions containing at least one cosmetic-acceptable excipient.

[0116] Within the scope of the present invention, the term “cosmetically acceptable” excipient means a compound and / or solvent, or equivalent thereof, that is cosmetically acceptable, particularly topically acceptable, i.e., does not induce unreasonable inflammation or allergic reactions upon contact with the skin, especially the scalp, is non-toxic, and is not unstable.

[0117] Within the scope of the present invention, the term “cosmetic composition” means a non-therapeutic composition, i.e., a composition intended for the prevention and / or care of the skin, in particular the scalp, and / or the skin's outer layer, in particular the hair, which a dermatologist would say is “normal,” i.e., non-pathological. With respect to the skin, scalp, skin's outer layer, or hair, the term “normal” is used herein to mean healthy skin, scalp, skin's outer layer, or hair as previously defined.

[0118] In a preferred embodiment of the present invention, the hydrolyzed extract according to the present invention is obtained in a quantity of 1 × 10¹⁶ based on the total weight of the composition. -4 Weight % to 10% of weight, 1x10 preferred -3 Weight % to 5% of weight, with 1x10 being preferred. -3 Weight % to 3% weight, and even more preferentially 1 x 10 -3 It is present in the cosmetic composition in a content of % to 1% by weight, preferably 0.01% to 1% by weight.

[0119] Therefore, the composition can be used to maintain and / or enhance the biomechanical properties of the skin, preferably keratin fibers, and more preferably hair.

[0120] The cosmetic compositions according to the present invention may be in presentation forms conventionally used for topical application to the skin or scalp and / or skin surface, preferably the scalp and / or skin surface, such as liquid or solid forms, or in pressurized liquid forms. They may be formulated in the form of aqueous or oily solutions, aqueous creams or gels or oily gels, especially jars or tubes, especially shower gels, shampoos, conditioners, milks, oils, emulsions, hydrogels, microemulsions or nanoemulsions, especially oil-in-water or water-in-oil or multiple or silicone emulsions, serums, especially lotions in glass or plastic bottles, measuring bottles, aerosols or sprays, vials, liquid soaps, pastes, skin bars, ointments, foams, masks, lacquers, patches, varnishes, or anhydrous products, preferably liquids, pastes or solids, and may be formulated, for example, in the form of wands, especially sticks, or in powder form. It may be a cosmetic product, particularly for eyelashes or eyebrows, such as mascara or pencil, or a makeup remover for eye care and beauty, and / or a product for nail care and beauty, and / or beard care and / or beauty. In particular, the cosmetic composition is selected from the group consisting of serums, lotions, creams, shampoos, hair conditioners, oils, milks, ointments, pastes, foams, emulsions, hydrogels, shower gels, masks, lacquers, sprays, waxes, mascaras, makeup pencils or varnishes, and more favorably, shampoos, hair conditioners or lotions.

[0121] Preferably, the hydrolyzed extracts according to the present invention are suitable for formulations of "neutral" compositions that are gentle on the skin, and more favorably on keratin fibers, especially hair fibers. The hydrolyzed extracts according to the present invention are also suitable for use in cationic formulations containing surfactants.

[0122] The cosmetic compositions according to the present invention may optionally contain any suitable solvent and / or any suitable vehicle and / or any suitable excipient in combination with other compounds of interest. These may contain, in particular, excipients that are acceptable as cosmetics, selected from surfactants, preservatives, buffers, swelling agents, chelating agents, biocides, denaturants, opacifiers, pH adjusters, reducing agents, stabilizers, emulsifiers, thickeners, gelling agents, film-forming polymers, solvents, fillers, bactericides, odor absorbers, gloss adjusters, conditioning agents, texturers, glossing agents, pigments, dyes, fragrances, chemical or mineral sunscreens, trace elements and essential oils. These combinations are also included in the present invention.

[0123] The cosmetic composition may contain other cosmetic agents having the same properties as the hydrolyzed extract according to the present invention, and in some cases may induce a synergistic effect with said hydrolyzed extract, or it may contain cosmetic agents having complementary effects such as hair loss inhibitors, hair protectants, sedatives, anti-aging agents, or anti-fouling agents. Examples of activators to combat hair loss include a combination of hair protectants such as sulfopeptides, amino acids, amino sugars, vitamin B, zinc, and extracts of Panax ginseng and Artium majus sold by the applicant under the name Trichogen® LS8960, or an extract of Litchi chinensis pericarp sold by the applicant under the name Litchiderm®, and sedatives and antipruritic agents such as rapeseed phytosterols sold by the applicant under the name Phytosoothe® LS9766.

[0124] Other activators, such as Cassia alata leaf extract sold under the name DN-Age (trademark) as an antioxidant activator, especially for hair care; a combination of Salvia miltiorhizza and niacinamide extracts sold under the name CollRepair (trademark) as a deglyceride; or activators that promote skin stiffness, and therefore scalp stiffness, such as synthetic tetrapeptide sold under the name Dermican (trademark); Hibiscus abelmoschus extract sold under the name Linefactor (trademark); refined pea extract sold under the name Proteasyl (trademark); Manilkara multinervis extract sold under the name Elestan (trademark); and Khaya senegalensis sold under the name Collalift (trademark) 18. The active ingredients may be present in compositions of extracts of Schizandra chinensis, argan pulp sold by the applicant under the name Argassential®, Schizandra chinensis sold under the name Sqisandryl®, Eperua falcata sold under the name Eperuline®, and Orthosiphon staminus sold by the applicant under the name MAT-XS® Bright. These combinations of active ingredients can, in particular, strengthen hair follicles and reduce hair loss. The hydrolyzed extracts of the present invention can also be combined with extracts of Nephelium lappaceum seeds, sold by the applicant under the name Rambuvital®, particularly for their hair-protective properties against contamination. The hydrolyzed extracts can also be used in combination with hydrolyzed extracts of Hippophae rhamnoides seed cake.

[0125] Advantageously, the cosmetic composition according to the present invention does not contain retinoids.

[0126] The third subject of the present invention relates to a non-therapeutic cosmetic care process comprising the topical application of a hydrolyzed extract according to the invention or a cosmetic composition comprising the same, for maintaining and / or increasing the biomechanical properties of the cutaneous cuticle, preferably keratin fibers, and preferentially hair.

[0127] In an advantageous embodiment of the present invention, the cosmetic care process consists of the topical application of a hydrolyzed extract according to the invention or a cosmetic composition comprising the same, to all or part of the skin of the body and / or face, including the cutaneous cuticle, preferably selected from the scalp, legs, thighs, arms, stomach, collar, neck, all or part of the face, forehead, chin, lip contour, eye contour, the area known as the "T-zone" of the face, and preferably the cutaneous cuticle selected from the scalp, and / or to all or part of the cutaneous cuticle, preferably to nails, hair, body hair, particularly beard, eyelashes and / or eyebrows, even more preferably to hair, preferentially to all or part of the keratin fibers, even more preferentially to all or part of the hair.

[0128] Thus, the cosmetic care process makes it possible to maintain and / or increase the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e. a non-brittle appearance, and / or the softness of the cutaneous cuticle, preferably keratin fibers, more preferably hair.

[0129] Another subject also relates to a cosmetic treatment method for maintaining and / or increasing the biomechanical properties of the cutaneous cuticle, preferably keratin fibers, and preferably hair, - identifying, in a human not suffering from lesions, particularly skin lesions, requiring a therapeutic treatment, the areas of the skin of the body and / or face, and / or zones of the cutaneous cuticle, including the cutaneous cuticle, preferably keratin fibers, and preferably hair, for which it is desired and / or necessary to maintain and / or increase the biomechanical properties thereof; - in particular, from 1×10 -4 % by weight to 10% by weight, preferably from 1×10 -3 % by weight to 5% by weight, even more preferably from 1×10 -3Weight % to 3% weight, and even more preferentially 1 x 10 -3 A step of topically applying a hydrolyzed extract or composition containing the same according to the present invention to the relevant area of ​​the skin of the body and / or face, including the skin surface, and / or the skin surface, in a content of % by weight to 1% by weight, preferably 0.01% by weight to 1% by weight, Includes.

[0130] The examples form an integral part of the present invention, and any features that appear novel to the prior art from the overall description, including the examples, also form an integral part of the present invention. Therefore, each example has a general scope.

[0131] Unless otherwise specified, temperature is expressed in Celsius and pressure is atmospheric pressure. [Examples]

[0132] Example 1: Different methods for preparing a hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention The cakes used in the following examples contain less than 8% by weight of fat relative to the total weight of the cake.

[0133] Example 1a: Enzyme hydrolysis - hydrolyzed extract 1 The milk thistle (Silybum marianum) seed cake was crushed and suspended in water (as the sole solvent) at a concentration of 15% by weight of the seed cake relative to the total weight of the cake and solvent. The mixture was subjected to hydrolysis at 65°C and pH 7.5 for 2 hours, maintaining the pH during hydrolysis, resulting in a weight concentration of 1.7% of the bacterial protease (Alcalase®) derived from the liquid form of Bacillus licheniformis strain relative to the 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 4.14% ± 0.3% by weight of dry material relative to the weight of the liquid product was obtained.

[0134] Example 1b: Enzyme hydrolysis - hydrolyzed extract 2 The milk thistle (Silybum marianum) seed cake was crushed and suspended in water (as the sole solvent) at a concentration of 12.5% ​​by weight of the seed cake relative to the total weight of the cake and solvent. The mixture was subjected to hydrolysis at 55°C and pH 8.5 for 1 hour, with the pH maintained during hydrolysis, and the weight concentration of the bacterial protease (Alcalase®) derived from the liquid form of Bacillus licheniformis strain relative to the weight of the cake was 1%. 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% by weight of dry material relative to the weight of the liquid product was obtained.

[0135] Example 1c: Hydrolyzed extract of cosmetic ingredient (containing maltodextrin) 1 The extract obtained in Example 1a) is atomized in the presence of maltodextrin, with a final amount of maltodextrin of 75% by weight relative to the total weight of the final atomized extract. Thus, the extract is obtained in powder form.

[0136] Example 1d: Hydrolyzed extract of cosmetic ingredient (containing maltodextrin) 2 The extract obtained in Example 1b) is atomized in the presence of maltodextrin, with a final amount of maltodextrin of 75% (by weight / by weight) relative to the total weight of the final atomized extract. Thus, the extract is obtained in powder form.

[0137] Example 1e: Enzymatic hydrolysis using hydrolyzed extract 1 from Example 1a The Silybum marianum seed cake was crushed and suspended in the hydrolyzed extract (solvent only) obtained in Example 1a) at a content 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 a temperature of 65°C and pH 7.5 for 2 hours, with the pH maintained during hydrolysis, and the weight concentration of the bacterial protein (Alcalase®) from the liquid form of Bacillus licheniformis strain relative to the weight of the cake was 1.7%. The enzyme was then inactivated by heating at a temperature of 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 material relative to the weight of the liquid product was obtained.

[0138] Example 2. Analysis of the peptide equivalent content and peptide molecular weight profile of the hydrolyzed extract from Example 1a in the form of a cosmetic ingredient as described in Example 1c. a) Total protein assay 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.

[0139] Materials and Methods: The peptide equivalent content of the hydrolyzed extract was estimated by performing a total nitrogen assay (Kjeldahl method - AOAC, Association of Official Analytical Chemists Official Methods of Analysis, AOAC, Washington, DC, USA, 2000) and multiplying the obtained value by 6.25 (N × 6.25). The results are summarized in Table 1.

[0140] [Table 1]

[0141] Results: The amount of dry product obtained after extraction and hydrolysis was 3.84% to 4.44%. Total peptides accounted for approximately 60% to 66.2% of the dry product of the hydrolyzed extract from Example 1a (Table 1).

[0142] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention is rich in plant peptides (60% to 66.2% by weight relative to the total weight of the dry material).

[0143] b) Analysis of the mean molecular weight profile of hydrolyzed peptides Molecular profile of hydrolyzed extract Objective: To characterize the weight-average molecular weight of the peptide compounds in the hydrolyzed extracts according to the present invention, two hydrolyzed extracts from Example 1a were studied and then classified into three categories: less than 100 kDa, 100 kDa to 20 kDa (including both ends), and greater than 20 kDa.

[0144] Materials and Methods: The weight-average molecular weight distribution of peptides in the hydrolyzed extract of cosmetic ingredients according to Example 1a was analyzed by gel permeation chromatography using a Superose® 12 10 / 300 GL column (GE Healthcare Life Sciences). The weight-average molecular weight was determined after calibrating the column with protein molecules of known weight-average molecular weight. The results are summarized in Table 2.

[0145] [Table 2]

[0146] Results: The peptides in the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention are mainly peptide compounds having a weight-average molecular weight of 100 Da to 20 kDa (Table 2).

[0147] Molecular profiles from 100 Da to 7000 Da Objective: The objective of this study was to investigate the weight-average molecular weight distribution profile of peptides contained in the hydrolyzed extract of Silybum marianum seed cake obtained according to Example 1c in the range of 100 Da to 7000 Da. Subsequently, the weight-average molecular weights were classified into three categories: two extreme categories combining the minimum molecular weight 100 Da < MW < 700 Da and the maximum molecular weight 2500 Da < MW < 7000 Da.

[0148] Materials and methods: The weight-average molecular weight distribution of peptides from the hydrolyzed extract of Silybum marianum 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). Fractions were taken in the range of 100 Da to 7000 Da, and UV detection was performed at 214 nm. The weight-average molecular weight was determined after calibrating the column with protein molecules of known weight-average molecular weight. The obtained results are summarized in Table 3.

[0149]

Table 3

[0150] Results: The hydrolyzed extract of Silybum marianum seed cake according to the present invention mainly (98.03%) has protein compounds with a low weight-average molecular weight in the range of 100 Da to 2500 Da (Table 3).

[0151] Example 3: In vitro effect of the hydrolyzed extract according to Example lb on the biomechanical properties of the skin epidermis - Free radical scavenging activity (DPPH) The objective of this study was to measure the free radical activity of the hydrolyzed extract according to Example lb using the DPPH method.

[0152] Materials and methods Diphenyl-1-picrylhydrazyl (DPPH°) is an oxidizing agent that is an electron-deficient free radical. In this form, the compound, which is dark purple in color, absorbs wavelengths from 510 nm to 530 nm. In the presence of an electron-donating reducing compound, diphenyl-2-picrylhydrazyl forms a stable compound with a decrease in its absorption properties (decolorization of the compound) (Non-Patent Literature 2). This method is used to select plant extracts that have free radical scavenging properties (Non-Patent Literature 3).

[0153] The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1b was mixed with distilled water at a 5% (w / v) concentration. 100 μl of this solution was tested in the presence of a similar volume of 90 μM diphenyl-1-picrylhydrazyl solution (DPPH°, 3.55 mg of DPPH powder in 100 mL of Sigma-ethanol). After incubation at room temperature for 30 minutes and protection from light, absorbance was recorded at 530 nm using a spectrometer (Victor V, Perkin Elmer). The results are expressed as a percentage of DPPH° free radical inhibition compared to a negative control.

[0154] Results: Summarized in Table 4. The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention preserved up to 95% of the oxidation of DPPH (Table 4).

[0155] [Table 4]

[0156] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention has strong free radical activity and is therefore effective in maintaining and / or increasing biomechanical properties.

[0157] Example 4: In vitro effect of hydrolyzed seed cake extract according to the present invention on the biomechanical properties of the skin surface - Antioxidant / Transition metal (Fe) chelating activity Glycation can occur chemically between reducing sugars and proteins. The presence of oxidizing agents and metals catalyzes this reaction and the formation of AGEs (Advanced Glycation End Products).

[0158] The chelating and antioxidant activity of the hydrolyzed extract of milk thistle (Silybum marianum) cake according to the present invention was evaluated in an in vitro model in the presence of albumin, iron, and an oxidizing agent, hydrogen peroxide. The oxidation reaction was evaluated by measuring the decomposition of tryptophan, tyrosine crosslinking, and the formation of glycation products including pentosidine.

[0159] Materials and methods A reagent containing 1.5% (w / v) albumin, 2 mM iron, 5 mM EDTA, and 25 mM hydrogen peroxide (H2O2) is brought into contact with, or not brought into contact with, the hydrolyzed extract of milk thistle (Silybum marianum) cake according to Example 1a, in a content of 0.003% or 0.01% by weight. Albumin is oxidized in the presence of H2O2. The oxidation reaction is catalyzed by iron. The catalytic activity of the oxidation reaction is controlled in the absence of iron. The mixture is incubated at 37°C for 1 day.

[0160] The amounts of tryptophan, dityrosine, pentosidine, and the fluorescent glycation products are measured by fluorescence using a Thermo Scientific Varioskan® Flash instrument (excitation / emission wavelengths: 280 nm / 340 nm for tryptophan; 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosidine; and 370 nm / 440 nm for the final glycation product).

[0161] The statistical significance threshold was set at 5% (p<0.05).

[0162] The protection index is calculated according to the following formula: (iron-containing control - treatment with hydrolyzed extract and iron according to the present invention) / (iron-containing control - iron-free control) × 100.

[0163] The test is performed on the extract from Example 1a, which has been dried to obtain a 100% dry extract content.

[0164] [Table 5]

[0165] The presence of iron catalyzes the oxidation reaction, which can be observed by the increased content of undegraded tryptophan in the iron-free albumin control (Table 5). In the presence of hydrolyzed extracts of milk thistle (Silybum marianum) seed cake according to Example 1a at 0.003% and 0.01% by weight, tryptophan degradation was significantly reduced, with 18% and 28% of tryptophan residues (undegraded) recovered, respectively. The protection index of the hydrolyzed extract of milk thistle (Silybum marianum) seed cake tested at 0.01% by weight was 38%.

[0166] [Table 6]

[0167] The presence of iron catalyzes the formation of dityrosine in albumin in the presence of H2O2 (Table 6). Hydrolyzed extracts of milk thistle (Silybum marianum) 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 (Silybum marianum) seed cake tested at 0.01% by weight was 67%.

[0168] [Table 7]

[0169] The presence of iron also catalyzes pentosidine formation in the presence of H2O2 (Table 7). A hydrolyzed extract of milk thistle (Silybum marianum) seed cake with 0.01% by weight of Example 1a reduces pentosidine formation by 27%. The protection index of the hydrolyzed extract of milk thistle (Silybum marianum) seed cake tested at 0.01% by weight is 61%.

[0170] [Table 8]

[0171] The presence of iron catalyzes the oxidation process of albumin in the presence of H2O2, leading to the formation of fluorescent glycation products (Table 8). Hydrolyzed extracts of milk thistle (Silybum marianum) cake according to Example 1a at 0.003% and 0.01% by weight reduced the formation of these glycation products by 9% and 14%, respectively. The protection indices of the hydrolyzed extracts of milk thistle (Silybum marianum) cake tested at 0.003% and 0.01% by weight were 18% and 27%, respectively.

[0172] Conclusion: Hydrolyzed extract of milk thistle (Silybum marianum) seed cake reduces the oxidation of iron-catalyzed proteins and the formation of glycation products, thus having an effect on maintaining biomechanical properties.

[0173] Example 5: In vitro effect of hydrolyzed seed cake extract according to the present invention on the biomechanical properties of the skin epidermis - Antioxidant / transition metal (Cu) chelating activity The objective of this study is to evaluate the antioxidant / metal chelating activity of the hydrolyzed extract of milk thistle (Silybum marianum) cake according to the present invention in in vitro tests in the presence of lipoproteins and copper.

[0174] Materials and methods Low-density lipoprotein (LDL) at 100 μg / mL is brought into contact with 200 μM copper in phosphate-buffered saline. A 0.048 wt% hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a is added to the solution, or not. The reaction mixture is incubated at 37°C for 48 hours.

[0175] The level of lipid oxidation is assessed by adding 2% by volume of thiobarbituric acid (TBA) to the total volume of the reaction mixture, followed by quantification of malondialdehyde (MDA). The fluorescence level is quantified by spectrophotometric analysis at an emission wavelength of 560 nm after excitation at a wavelength of 532 nm using a Thermo Scientific Varioskan® Flash instrument.

[0176] The Schiff base content is evaluated by spectroscopy using a Thermo Scientific Varioskan® Flash instrument (excitation wavelength 370 nm / emission wavelength 440 nm).

[0177] The statistical significance threshold was set at 5% (p<0.05).

[0178] The protection index is calculated according to the following formula: (treatment with hydrolytic extract according to the present invention and LDL+copper-LDL+copper control) / (control without copper - LDL+copper control) × 100.

[0179] Results: The results are summarized in Table 9 below.

[0180] [Table 9]

[0181] The presence of copper enhanced the oxidation of lipids assayed with malondialdehyde. Hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a, used at 0.048 wt%, significantly reduced malondialdehyde formation by 78% (Table 9). The protection index of hydrolyzed extract of milk thistle (Silybum marianum) cake according to Example 1a, tested at 0.048 wt%, was 97.5%.

[0182] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention reduced copper-catalyzed lipid oxidation, thus maintaining and / or increasing biomechanical properties.

[0183] Example 6: In vitro effect of hydrolyzed seed cake extract according to the present invention on the biomechanical properties of the skin epidermis - Anti-glycation activity on hair fiber proteins Objective: The objective of this study is to measure the anti-glycation effect of hydrolyzed extracts of milk thistle (Silybum marianum) seed cake according to Example 1a on hair fiber proteins incubated in glucose solution.

[0184] Materials and methods Standardized and washed hair bundles are subjected to 1 mol.L of a solution containing or not containing the hydrolyzed extract of Example 1a. -1 The rocks were incubated in (M) phosphate-buffered glucose solution and dried at 50°C for 2 days at pH 7.4 to obtain a 100% dry extract with a content of 0.01% by weight. The rocks were then rinsed, squeezed, dried, combed, separated, and cut into 1 cm long pieces. The proteins were extracted in 1N sodium hydroxide (NaOH) solution at room temperature for 24 hours with gentle stirring.

[0185] The amounts of dityrosine and pentosidine (glycation products) are measured by fluorescence using a Thermo Scientific Varioskan® Flash instrument (excitation / emission wavelengths: 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosidine).

[0186] The protection index is calculated according to the following formula: (Saccharification control - Saccharification + treatment with the extract according to the present invention) / (Non-saccharification control - Saccharification control) × 100.

[0187] The statistical significance threshold was set at 5% (p<0.05).

[0188] Results: These are shown in Tables 10 and 11 below.

[0189] [Table 10]

[0190] Incubation of fibers in glucose solution increased the dityrosine content in hair proteins. A hydrolyzed extract of milk thistle (Silybum marianum) seed cake, used at 0.01% by weight in Example 1a, significantly reduced dityrosine formation by 16% (Table 10). The protection index of the hydrolyzed extract from Example 1a, used at 0.01% by weight, against dityrosine formation was 57%.

[0191] [Table 11]

[0192] Incubation of fibers in glucose solution increased the pentosidine content in hair proteins. A hydrolyzed extract of milk thistle (Silybum marianum) seed cake, used at 0.04% by weight in Example 1a, significantly reduced pentosidine formation by 17% (Table 11). The protection index of the hydrolyzed extract from Example 1a, used at 0.04% by weight, against dityrosine formation was 59%.

[0193] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention reduces the formation of glycation products in the presence of glucose in hair fibers, and thus makes it possible to maintain and / or increase the biomechanical properties of hair fibers.

[0194] Example 7: Ex vivo effect of hydrolyzed extract according to the present invention on the biomechanical properties of the skin epidermis - Antioxidant / carbonylation activity on keratin Objective: The objective of this study was to measure the protective effect of the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention against oxidation, particularly the carbonylation of keratin and keratin-related proteins (KAPs).

[0195] Materials and methods Preparation of hair locks: Hair braids from Caucasian donors were used for measurement. The hair was treated by contacting it with a 0.05% (w / v) hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a in distilled water for 10 minutes, followed by a washing and drying process. Hair fibers (PM; ERM-CZ100 fine powder, particulate matter 10-like (PM10-like), 170 μg / cm³ 2 Urban particles (fine particles) are applied to polycyclic hydrocarbon aromatics, and immediately irradiated with UVA (84 J / cm²). 2 Stressed hair was subjected to exposure for 6 hours using an LED light source, with peak emission at 365 nm. Hair treated with water or 0.2% (w / v) N-acetylcysteine ​​(antioxidant - positive control) and exposed or unexposed to UVA and particles was used as a control.

[0196] Hair shafts were sampled from each experimental condition, cryopreserved, frozen in liquid nitrogen, and kept at -80°C until analysis.

[0197] a) Quantification of carbonylated proteins by Western blotting Materials and methods Carbonylated proteins in hair shafts were quantified using Western blot analysis under various conditions.

[0198] Total proteins (keratin and keratin-related proteins) were extracted from each hair shaft using a process conventionally known to those skilled in the art. Total protein quantification was performed using the Bradford method. Carbonylated proteins were labeled with a specific fluorescent probe (Ex=647nm / Em=650nm; Non-Patent Literature 4). Proteins were separated by SDS-PAGE, and their fluorescence signals were quantified by acquiring gel images using the ThermoFisher "iBright" software system. Total proteins were stained in the gel for normalization using the "iBright" system (ThermoFisher). A carbonyl score value (carbonylated protein / total protein), representing the quantification of carbonylated proteins, was obtained for each sample. Carbonyl score (Sample X) = Fluorescence value of oxidized protein (Sample X) / Fluorescence value of total protein (Sample X)

[0199] The protection index (%) was calculated using the following formula. Protection Index (%) = (Carbonyl Score Level (Stress) - Carbonyl Score Level (Treatment with Extract According to the Present Invention)) / (Carbonyl Score Level (Stress) - Carbonyl Score Level (Control)) × 100

[0200] For reference, the control group is thought to have maximum efficacy (100%), while the stress group has minimum efficacy (0%).

[0201] The statistical significance threshold was set at 5% (p<0.05).

[0202] Results: The results are summarized in Table 12 below.

[0203] [Table 12]

[0204] UVA and pollution (UVA 84J / cm²) 2 A significant increase in protein carbonylation was observed during exposure to stress caused by (and urban PM10 particles) (Table 12).

[0205] A hydrolyzed extract of milk thistle (Silybum marianum) seed cake at 0.05% (w / v) in Example 1a significantly reduced the level of carbonyl proteins in hair fibers by 25% (p<0.01). This corresponds to a protection index >100%.

[0206] b) Quantification of carbonylated proteins by image analysis Materials and methods In-situ concentration analysis of carbonylation signals was performed by analyzing images obtained using ThermoFisher's "iBright" software and processed with Rasband's ImageJ software. The oxidation levels of each experimental group in the cuticle and cortical regions are shown as mean (UFR) ± standard deviation of the mean (Tables 13 and 14).

[0207] The statistical significance threshold was set at 5% (p<0.05).

[0208] [Table 13]

[0209] Result: 84J / cm 2 UVA and urban particles increased the amount of carbonyl proteins in the cortex and cuticle.

[0210] [Table 14]

[0211] A hydrolyzed extract of milk thistle (Silybum marianum) seed cake in Example 1a at a concentration of 0.05% (w / v) significantly reduced the carbonyl protein content in the cuticle and cortex by 28% (p<0.01) and 31% (p<0.001), respectively, thus maintaining and / or increasing the biomechanical properties of the hair. This corresponds to a protective index of 49% for the cuticle and 59% for the cortex.

[0212] c) Protection of the structural integrity of hair Objective: To evaluate the structural state of keratin protein in hair shafts obtained according to the above protocol by optical measurement using an XPolar® microscope (KAMAX). The birefringence properties of hair fibers depend on the structural integrity of the keratin.

[0213] Hair is composed of fibrous keratin protein. The crystalline structure of keratin fibers gives hair a property known as birefringence, which is the ability to correct the polarization of light passing through the hair. Birefringence depends on the hair thickness and the crystalline state of the keratin. A decrease in the birefringence parameter (Kindex) indicates structurally damaged keratin. The Kindex value related to the birefringence of hair fibers can be determined and compared between damaged hair and undamaged hair, treated hair and untreated hair.

[0214] Materials and methods Hair segments 1 cm in length (n=30 per condition) obtained according to the protocol described above were transferred to slides adapted for birefringence analysis using XPolar technology. This is an imaging device integrated into a microscope, allowing observation and measurement of the polarization properties of the investigated sample. Reflectance images were first taken to measure the hair thickness. Then, the sample was irradiated with polarized light and the Kmax was measured. Assuming a cylindrical hair shape, the birefringence value (Kindex) can be estimated from the Kmax value and hair diameter using an abacus.

[0215] The value for improvement in hair structure was obtained using the damaged group and the untreated group as references, according to the formula: (stressed hair treated with the hydrolyzed extract according to the present invention - stressed hair) / (healthy hair - stressed hair) × 100.

[0216] [Table 15]

[0217] Results: Oxidative stress caused structural changes in hair fibers. A decrease in Kindex values ​​was observed in damaged hair compared to undamaged control hair (Table 15). This corresponds to the degradation of keratin structure due to oxidation of keratin under exposure to UVA and urban particles.

[0218] Hair fibers treated with a hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a at 0.05% (w / v) in distilled water showed a significant increase in mean birefringence compared to damaged hair fibers (Table 15). The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a enabled a 14% (p<0.05) increase in birefringence. This corresponds to a 59% improvement in hair structure (improvement index) compared to damaged hair, and therefore corresponds to the improvement and / or maintenance of the biomechanical properties of the hair fibers.

[0219] d) Integrated Hair Protection Score The Hair Protection Score (HPS) is an integrated index for classifying the effectiveness of hair care products. Score values ​​are distributed between: HPS15; HPS30; HPS50; and HPS50+. HPS values ​​are obtained using a specific algorithm that integrates initial molecular events to assess the protection of hair fibers against daily damage. The HPS of the product was obtained by integrating molecular damage, assessed by evaluating protein carbonylation, and structural damage, assessed by evaluating hair birefringence according to Example 7, supporting effective protection of hair fibers against damage induced by contamination (particulate and UVA irradiation). An internal standard was used as a control (N-acetylcysteine: HPS=50).

[0220] The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a showed a hair protection score of 50+. This score is higher than that of N-acetylcysteine.

[0221] Statistics: All data were analyzed using GraphPad Prism 9 (Insightful Science USA) or SigmaPlot (Systat Software Inv. USA) software. Statistical analysis was performed using binomial Student t-tests or ANOVA comparisons between the injury group and the condition. The statistical significance threshold was set at 5% (p<0.05).

[0222] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention protects keratin and keratin-related proteins from carbonylation induced by UV and pollutant exposure of hair fibers, enabling the maintenance of their structural integrity and biomechanical properties.

[0223] Example 8: Penetration (FT-IR) (Exvivo) of hydrolyzed extract into fibers (cortex) on hair fibers. Objective: 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 various environmental and treatment conditions. These techniques are non-destructive and do not require sample labeling. FT-IR is particularly advantageous for the analysis of colored hair because it is less affected by the strong absorption of melanin under visible excitation. The objective of this study is to evaluate the distribution of components of the hydrolyzed extract according to the present invention in hair fibers and to study its effect on the infrared profile of keratin.

[0224] Materials and methods Hair Preparation and Treatment: The test was performed on three Caucasian human hair strands decolorized with persulfate (super decolorization). Before the test, hair locks were normalized (2g / 2g / 15cm), washed, rinsed, and dried at room temperature. Braids of healthy Caucasian hair (natural dark brown hair) were used as a control for natural, undamaged hair. The hydrolyzed extract according to Example 1a was mixed with 5% w / v distilled water or water as a placebo and completely covered the hair locks. Excess product was removed from the locks by wiping each strand between a gloved thumb and a gloved index finger. The hair was cut into 10 μm sections for infrared imaging.

[0225] Infrared spectroscopy: The vibrational spectral profile of the product was characterized by infrared spectroscopy combined with microscopy. The image is 6.25 × 6.25 μm. 2 With a pixel size of 800-4000cm -1 The samples were collected between [specific time periods]. The spectra were washed after pretreatment for analysis. The product spectra were tracked on hair fibers using a fitting method.

[0226] Results: The hydrolyzed extract from Example 1a had different keratin spectra at 993, 1053, 1110, 1403, and 1597 cm⁻¹. -1It possesses specific spectral bands. These spectral markers were used to identify hydrolyzed extracts in hair fibers. The hydrolyzed extracts were observed to be concentrated primarily in the hair cuticle and diffuse into the cortex of the fiber.

[0227] Example 9: Effects of hydrolyzed extract according to the present invention on the biomechanical properties of the skin epidermis - Molecular reconstruction and stabilization of cysteine ​​groups (FT-IR) in hair fibers (exvivo) Objective: The objective is to more accurately study the vibrational spectrum of keratin, the presence or absence of damage, and whether or not it was treated with the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a. These analyses demonstrated the interaction between the hydrolyzed extract according to the present invention and damaged keratin in the hair cortex, particularly its general stabilizing effect, and especially its effect on cysteic acid groups and their derivatives.

[0228] Materials and methods Brown Caucasian hair locks were oxidized by bleaching, calibrated (12 cm / 1 g), washed, and dried for study. The locks were bleached with a 5% v / v hydrogen peroxide solution for 20 minutes, then rinsed and dried.

[0229] Comparative analysis of keratin vibration spectra from undamaged hair and bleached and oxidized hair treated with or without the extract of Example 1a mixed in distilled water at 5% w / v was performed using a "fitted" analysis: a programmed method comparing the infrared spectra of a given control with those of the product using a Matlab environment. Changes in the keratin signal were processed using hierarchical cluster analysis to determine the similarity between spectra. The area under the curve was 1174 cm². -1 and 1116cm -1 The absorption band was determined.

[0230] When comparing values, the statistical significance threshold was set to 5% (p<0.05).

[0231] result: Oxidative damage caused by chemical decolorization of hair showed effects on the chemical and structural composition of the hair, accompanied by observable changes in the vibrational spectral profile.

[0232] Hierarchical cluster analysis showed that damaged hair and virgin hair treated with the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a had spectral profiles closer to those of damaged hair than to those similar. These results suggest that the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention interacts with damaged keratin in the fibrous cortex, stabilizing it.

[0233] Next, the spectral bands relating to the integrity of cysteine ​​bonds were studied in more detail. These were modified under conditions of chemical oxidation compared to healthy, undamaged hair.

[0234] In particular, 840, 1116, 1150 and 1174 cm -1 The spectral bands (referring to the stretching of NH, SO2, and CO outside the bending plane, and the asymmetric stretching of SO3, respectively) showed increased intensity in hair oxidized by hair bleaching compared to healthy hair.

[0235] It has been demonstrated that the hydrolyzed extract according to the present invention can reduce these band intensities toward the band intensities of virgin hair.

[0236] Finally, it was demonstrated that the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention can reduce these band strengths to levels closer to those of healthy hair (Table 16).

[0237] Specifically, oxidative damage acted on the SO, SO2, NH, and CO bonds (a significant increase in the intensity of these bands). These groups were created by oxidation of the groups and cleavage of the cysteine disulfide bridges. The hydrolyzed extract of Silybum marianum seed cake according to the present invention showed a significant decrease in the intensity of these SO, SO2, NH, and CO bonds.

[0238] The protection index is calculated according to the following formula: (glycation control - glycation by the extract according to the present invention + treatment) / (non - glycated control - glycation control) × 100.

[0239] [[ID=...]]

Table 16

[0240] Conclusion: The hydrolyzed extract of Silybum marianum seed cake according to the present invention can interact with the molecular species of keratin and keratin - related proteins in hair fibers damaged by chemical oxidation and stabilize them. The hydrolyzed extract of Silybum marianum seed cake according to the present invention has a structure - restoring effect and thus a biomechanical property, particularly an effect in the restoration and / or maintenance and / or increase of strength, plasticity, resistance, and / or flexibility of 14% - 98% depending on the chemical groups of the bands between 840 - 1174 cm -1 This enables the restoration and / or maintenance and / or increase of strength, plasticity, resistance, and / or flexibility of 14% - 98% depending on the chemical groups of the bands between 840 - 1174 cm.

[0241] Example 10: Ex vivo effect of the hydrolyzed extract according to the present invention on the repair of hair fibers The purpose of this study is to measure the repair effect of the hydrolyzed extract of Silybum marianum seed cake according to the present invention on hair fibers damaged by hydrogen peroxide by calorimetry (DSC = differential scanning calorimetry).

[0242] Materials and methods Dark brown Caucasian hair locks were calibrated (1 g; 12 cm) and prepared for study. The hair locks were washed and then decolorized three times for 30 minutes each with a hydrogen peroxide solution (5.6% H2O2 by volume + 13.9% (NH4)2S2O8 by volume, pH=9.4). Healthy (undecolorized) hair locks were maintained as a control. After washing and drying under running air at 55°C for 45 minutes, the hair locks were immersed for 24 hours in distilled water (as a control) or aqueous solution containing a 1% (w / v) hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1b. Both solutions were pre-buffered at pH 5.5. The locks were rinsed and then dried under running air at approximately 60°C for 1 hour.

[0243] Following the method described in (Non-Patent Literature 5), the denaturation temperature of human protein was determined by differential calorimetry using a differential enthalpy analyzer (DSC Q100, TA Instruments) at a heating rate of 2 K / min.

[0244] The repair index relative to placebo is calculated as follows: (Decolorized hair treated with hydrolyzed extract according to the present invention - control + water (placebo)) / (Control healthy hair - control + water (placebo)) × 100.

[0245] The repair index relative to placebo is calculated as follows: (Decolorized hair treated with the hydrolyzed extract according to the present invention - control decolorized hair) / (control decolorized hair - control + water (placebo)) × 100.

[0246] [Table 17]

[0247] result: Hair bleached with hydrogen peroxide is more unstable than healthy, unbleached hair (Table 17). The difference in denaturation temperature of hair fiber proteins between damaged and healthy hair was -9.6°C.

[0248] Treatment of damaged hair with a 1% (w / v) extract of milk thistle (Silybum marianum) seed cake in Example 1b allowed for a +5.4°C improvement in hair protein stability against heat denaturation compared to damaged hair treated with water. The hydrolyzed extract of milk thistle (Silybum marianum) seed cake in Example 1b showed a restorative effect on hair fibers damaged by chemical oxidative stress compared to 64% of healthy hair.

[0249] Conclusion: Treatment of peroxide-treated hair with the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention enabled improvement of the stability of hair fiber proteins against heat. The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention demonstrated a restorative effect on hair fibers damaged by hydrogen peroxide (oxidative stress), and therefore showed effectiveness in restoring and / or maintaining and / or increasing biomechanical properties, particularly the strength, plasticity, resistance and / or flexibility of the hair fibers.

[0250] The efficacy of a 1% (w / v) hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention in repairing the cortex is 65% compared to a damaged control treated with water (placebo) and 69% compared to a damaged untreated control.

[0251] Example 11: Ex vivo effect of the extract according to the present invention on strengthening hair fibers (single fiber fatigue test) Objective: The effect of Example 1a on the strengthening of brittle hair fibers was evaluated by fatigue testing of isolated hair fibers. Fatigue testing assesses the tendency of hair to break under repeated force application (Non-Patent Literature 6). The mechanical tensile properties of hair reflect the internal state of its structure. This method can be considered a more realistic simulation of consumer practices where grooming represents such stress. This method also allows for the evaluation of the restorative effects of cosmetic ingredients, particularly on the cuticle.

[0252] Materials and Methods Dark brown Caucasian hair locks bleached three times with hydrogen peroxide were calibrated (1 g / 12 cm) and washed for the study. The locks were treated or not treated for 1 hour at pH 5.0 - 5.5 with an extract of Silybum marianum seed cake according to Example 1a mixed with distilled water, then rinsed and dried. Fifty to sixty fibers were tested for each treatment condition or non - treated condition under standardized conditions of 21°C ± 2°C and 40% ± 5% relative humidity.

[0253] In this test, individual hair fibers were repeatedly stressed. The instrument counts the number of cycles of a given stimulus necessary to induce breakage. The maximum number of cycles was defined. Under these test conditions, the fibers may or may not break during the experiment. The Kaplan - Meyer survival curve principle was used to calculate the probability of survival at a given time point to estimate survival (or non - broken keratin fibers).

[0254] Statistical analysis to compare groups was performed using a non - parametric model. Differences were considered significant for values of p < 0.05.

[0255] The hair strengthening index was calculated using the following formula: (the number of this cycle before breakage of the treated bleached hair - the number of this cycle before breakage of the untreated bleached hair) / (the number of this cycle before breakage of healthy hair - the number of this cycle before breakage of the untreated bleached hair)×100.

[0256]

Table 18

[0259] Conclusion: The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention has properties that strengthen and therefore repair hair fibers, enabling a reduction in the probability of hair fiber breakage, and thus maintaining and / or increasing the strength and / or plasticity and / or resistance and / or flexibility of hair fibers, and repairing the skin and, preferentially, hair.

[0260] Example 12: Ex vivo effect of hydrolyzed extract according to the present invention on the biomechanical properties of hair fibers - color preservation (colorimetric) Objective: The antioxidant and color-protective effects of the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention were evaluated in a hair dyeing test after exposure to sunlight.

[0261] Materials and methods Caucasian hair locks, decolorized twice with hydrogen peroxide and dyed with red pigment, were used. For the study, the hair locks were calibrated (1 g / 15 cm), washed, and dried. The locks were treated for 1 hour with a 1% (w / v) extract of milk thistle (Silybum marianum) seed cake according to Example 1a, mixed with distilled water, or left untreated, and then dried overnight at room temperature. The locks were then subjected to a 592 W / m² test. 2 The subjects were exposed to sunlight (Suntest CPS+) with wavelengths of 300-800 nm for 19 hours at a lamp output of [specified value]. The total exposure energy was 41469 kJ / m³ per cycle. 2The rocks were then washed with water for 3 minutes after exposure, dried overnight in a chamber adjusted to 40% relative humidity and 21°C, and subsequently colorimetrically measured using the L*a*b* method. The parameters measured were Delta E* (overall color change) and Delta a* (red change), which relates to the red / green hue. The color of the rocks was measured after 1, 2, or 3 cycles of treatment and exposure to sunlight. Eight rocks were used for each condition.

[0262] Statistical analysis of the data was performed by comparing the groups using Student's t-test. Differences with a p-value of <0.05 were considered statistically significant.

[0263] Using the stressed dyed group and the untreated group as reference, the percentage of effectiveness in preserving hair color against sun-induced decolorization was obtained according to the formula: 100 - [(dyed hair treated with hydrolyzed extract according to the present invention / control dyed hair) × 100].

[0264] [Table 19]

[0265] result: Dyed hair exposed to sunlight showed increased overall color and redness changes during the exposure cycle (Tables 19 and 20). Exposure to sunlight bleached the hair.

[0266] Treatment of dyed hair with hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a significantly slowed these color changes (delta a* and delta E*) as a function of the exposure cycle.

[0267] The hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention has a protective effect on hair color against sun-induced decolorization, and therefore makes it possible to maintain and / or increase the color and / or shine of hair fibers.

[0268] Example 13: Ex vivo effect of hydrolyzed extract according to the present invention on the biomechanical properties of hair - prevention of damage Objective: The effectiveness of hair treatments in preventing breakage and split ends was tested using a device that allows for repeated combing of hair strands. This allows for the investigation of the effects of chemical hair treatments such as bleaching on hair breakage. Subsequently, the protective effects of commercially available reference products and the hydrolyzed extract according to the present invention were studied.

[0269] Materials and methods For the study, Caucasian brown hair locks were calibrated (12 cm / g), washed, and dried. The locks were decolorized with a 5% v / v hydrogen peroxide solution for 20 minutes, then rinsed and dried. After treating with a shampoo of the following composition (equivalent to 0.25 g shampoo / 1 g hair) containing the hydrolyzed extract according to Example 1c in the form of a cosmetic material, the locks were treated with a hair conditioner of the following composition (equivalent to 0.125 g hair conditioner / 1 g hair) containing the hydrolyzed extract according to Example 1c in the form of a cosmetic. The product was rinsed, and the locks were left to stand at 40% relative humidity for 5 hours before analysis. The locks were styled 50,000 times under the following conditions: 30°C and 40% relative humidity. Broken fibers less than 9 cm in length were collected and weighed.

[0270] The breakage value was calculated using the formula: Breakage (g) / Lock (g) × 100.

[0271] Outliers were detected using the Grubbs test at a significance level of 0.05 and excluded from the data record.

[0272] Contains damage-preventing shampoo:

[0273] [Table 20]

[0274] Hair conditioner formulation to prevent damage:

[0275] [Table 21]

[0276] [Table 22]

[0277] Results: The percentage of breakage in untreated brittle hair was estimated to be 20.1%, compared to 4.5% in brittle hair treated with shampoo and hair conditioner containing hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1c (Table 22). The protection against breakage of the final product containing hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1c was 78%.

[0278] Conclusion: Using the shampoo and hair conditioner containing the hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention, good conditioning properties and protection of hair against damage were found.

[0279] Example 14: In vitro effects of hydrolyzed extract according to the present invention on biomechanical properties - protection and stimulation of metabolism on the papilla of hair follicles Papillary fibroblasts play a crucial role in regulating the regulatory signals involved in hair follicle cell metabolism (Non-Patent Literature 7; Non-Patent Literature 8). With aging or in response to environmental stress, the physiological function and metabolism of papillary fibroblasts may be affected, potentially impacting the quality of the resulting keratin fibers.

[0280] Objective: The objective of this study was to evaluate the anti-aging and metabolic stimulating effects of hydrolyzed extract of milk thistle (Silybum marianum) seed cake, as described in Example 1a, on a culture model of hair follicle-derived papillary fibroblast aggregates by measuring ATP. This three-dimensional cell model allows for the in vivo preservation of papillary characteristics, particularly its ability to induce the formation and growth of high-quality hair (Non-Patent Literature 9).

[0281] Materials and methods A suspension of fibroblasts (40 × 10³ cells) from human hair follicle papillae in DMEM, containing 0.2% v / v growth factor (mesenchymal stem cell proliferation supplement) and with or without 0.01% by weight / volume of hydrolyzed extract of milk thistle (Silybum marianum) according to Example 1a in distilled water, was placed in a 96-well microplate and centrifuged at 200 × g for 5 minutes to form aggregates. The aggregates were incubated at 37°C in a controlled atmosphere (5% CO2 and 95% relative humidity).

[0282] For the analysis of the target parameters, the aggregates were rinsed with 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.

[0283] Using cells in suspension, ATP was measured by bioluminescence according to the supplier's instructions (Bioluminescence Assay Kit CLS II Roche 11699695001, Sigma-Aldrich).

[0284] Results: Treatment of aggregates with a hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to Example 1a at 0.01 wt / vol% increased the total amount of ATP produced (+32% compared to control; Table 23).

[0285] [Table 23]

[0286] Conclusion: Treatment of papillary fibroblast aggregates with hydrolyzed extract of milk thistle (Silybum marianum) seed cake according to the present invention enables an increase in ATP within the pseudopapilla. These results suggest activation ability on hair follicles for structural and functional enhancement of keratin fibers.

[0287] Example 15: Consumer testing using shampoo and hair conditioner containing hydrolyzed extracts of cosmetic ingredients in the form prescribed in Example 1c. Objective: The purpose of this study is to evaluate consumer perception of the benefits provided to damaged hair by hydrolyzed extracts of cosmetic ingredients incorporated into shampoos and hair conditioners.

[0288] Materials and methods This study was conducted under double-blind, randomized conditions with a total of 31 volunteers. The shampoo and conditioner used were the same as those used in Example 13 (Tables 20 and 21).

[0289] The same volunteers applied either a shampoo or hair conditioner containing a hydrolyzed extract of a cosmetic ingredient or a placebo product at least three times a week over a four-week period, with a three-week treatment break between the two treatment stages.

[0290] Volunteers completed a questionnaire regarding their perception of consumer benefits, and their opinions on the product after four weeks of use were measured. The number of positive or negative responses was grouped and analyzed using a nonparametric binomial test (Non-Patent Literature 10). The significance threshold was set at 5% (p<0.05).

[0291] result: Regular use of shampoos and hair conditioners containing hydrolyzed extracts as cosmetic ingredients resulted in a majority of volunteers experiencing beneficial effects on their hair. Among these positive comments, the use of this hygiene and care routine was recognized as providing protection against chemical stress, breakage, and split ends. Hair appeared less damaged and healthier. Hair showed more vitality, volume, and shine, and appeared less dry. The scalp appeared more comfortable.

[0292] In contrast, placebo shampoos and hair conditioners that did not contain hydrolyzed extracts as cosmetic ingredients did not show a majority of favorable opinions regarding the perceived benefits of protecting hair from chemical stress, split ends, increased vitality, increased hair volume, and reduced dryness.

[0293] [Table 24]

[0294] Conclusion: Further benefits of the presence of hydrolyzed extracts as cosmetic ingredients on hair were recognized by volunteers who regularly used shampoo and hair conditioner for four weeks.

[0295] Example 16: Cosmetic ingredients Cosmetic ingredients 1 (weight %): Hydrolyzed extract 20% according to Example 1a) Maltodextrin 80% Cosmetic ingredients 2 (by weight): Hydrolyzed extract 30% according to Example 1b) Maltodextrin 70%

[0296] Example 17: Cosmetic composition containing cosmetic ingredients Damage prevention shampoo formulation

[0297] [Table 25]

[0298] Hair conditioner formulation to prevent damage

[0299] [Table 26]

Claims

1. Non-therapeutic cosmetic use of hydrolyzed extract of milk thistle (Silybum marianum) seed cake to maintain and / or increase the biomechanical properties of the skin's outer layer, advantageously keratin fibers, and preferably hair.

2. The use according to claim 1, wherein the hydrolyzed extract is obtained in water as the sole solvent.

3. The use according to claim 1 or 2, wherein the hydrolyzed extract is obtained by enzymatic hydrolysis.

4. The use according to claim 3, wherein the hydrolyzed extract is obtained by enzymatic hydrolysis at a pH of 3 to 10, preferably 7 to 9.

5. The use according to any one of claims 1 to 4, wherein the hydrolyzed extract has a total protein content of 50% to 95% by weight, preferably 60% to 90% by weight, and more preferably 60% to 70% by weight, based on the total dry mass of the hydrolyzed extract.

6. The use according to any one of claims 1 to 5, 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 50% to 90% by weight, preferably 65% ​​to 90% by weight, and more preferably 75% to 90% by weight, relative to the total dry mass of peptides in the hydrolyzed extract.

7. The use according to any one of claims 1 to 6, wherein the hydrolyzed extract has a dry mass content of peptides having a weight-average molecular weight of 100 Da to 700 Da of 60% to 90%, preferably 70% to 80%, and more preferably 75% to 80%, relative to the total dry mass of peptides having a weight-average molecular weight of 100 Da to 7000 Da of the hydrolyzed extract.

8. The use according to any one of claims 1 to 7, wherein the hydrolyzed extract maintains and / or increases the resistance and / or strength and / or volume and / or color and / or gloss and / or plasticity, i.e., non-brittle appearance and / or flexibility, of the skin exoskeleton, preferably keratin fibers, and advantageously the hair.

9. The use of the hydrolyzed extract according to any one of claims 1 to 8, wherein the hydrolyzed extract is intended for topical application to all or part of the skin of the body and / or face, including the skin, particularly the scalp, and / or all or part of the skin, preferably to the nails, hair, body hair, particularly the beard, eyelashes and / or eyebrows, more preferably to all or part of the keratin fibers, and more preferably to all or part of the hair.

10. The use according to claim 9, wherein the application is made to damaged, dyed, or highlighted hair, dull, dry, brittle, fragile, easily broken, thin, split ends, and / or stressed hair.

11. The use according to any one of claims 1 to 10, wherein the hydrolyzed extract is atomized in the presence of maltodextrin at a weight concentration of 1% to 99%, preferably 5% to 90%, more preferably 40% to 80%, and more preferably 70% to 80%, relative to the total weight of the resulting powder.

12. The hydrolyzed extract is present in an amount of 1 × 10⁻¹⁶ relative to the total weight of the cosmetic composition. －４ Weight % to 10% weight, preferably 1 x 10 －３ % to 5% by weight, with a higher preference of 1 x 10 －３ Weight % to 3% by weight, and even more preferably 1 x 10 －３ The use according to any one of claims 1 to 11, wherein the substance is present in the cosmetic composition in an amount of weight % to 1% by weight, preferably 0.01% to 1% by weight.

13. The use according to claim 12, wherein the cosmetic composition comprises at least one excipient that is acceptable as a cosmetic, and the composition is a serum, lotion, cream, shampoo, hair conditioner, oil, milk, ointment, paste, foam, emulsion, hydrogel, shower gel, mask, lacquer, spray, wax, mascara, makeup pencil, or varnish, preferably selected from shampoo, hair conditioner, or lotion.

14. The use according to claim 1 for repairing the skin crust, and more preferably the hair.

15. A non-therapeutic cosmetic care process comprising topical application of a hydrolyzed extract of milk thistle (Silybum marianum) seed cake or a cosmetic composition containing the same for maintaining and / or increasing the biomechanical properties of the skin exocutaneous, preferably keratin fibers, and preferably hair.

16. The cosmetic care process according to claim 15 for repairing the skin crust, preferably keratin fibers, and more preferably the hair.

17. The cosmetic care process according to claim 15 or 16, wherein the hydrolyzed extract or the cosmetic composition containing the same is applied topically to all or part of the skin of the body and / or face, particularly including the scalp, and / or to all or part of the skin, preferably to all or part of the keratin fibers, and more preferably to all or part of the hair.

18. The cosmetic care process 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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