Cosmetic use of the hydrolysate of Hippophae rhamnoides cake

Abstract

The present invention relates to the cosmetic use of a hydrolysate of the cake of Hippophae rhamnoides for increasing the growth and / or reducing its loss of skin appendages; for increasing their biomechanical properties and / or their superficial and / or histological properties; and / or for repairing damaged skin appendages. The present invention also relates to a cosmetic care method comprising the topical application of said hydrolysate of the cake or a cosmetic composition comprising it. The present invention also relates to a hydrolysate of the cake of H. rhamnoides or a dermatological or pharmaceutical composition comprising it for dermatological or pharmaceutical use in the treatment of alopecia and / or alopecia.

Description

[Technical field]

[0001] The present invention relates to a novel cosmetic use of a hydrolysate of the cake of Hippophae rhamnoides. [Background technology]

[0002] The care of skin appendages, such as hair, eyelashes and eyebrows, involves not only a global treatment, i.e. a direct treatment of the hair or keratin fibres, but also the treatment of the corresponding skin area, including the hair follicles. In the case of hair, the follicles are distributed over the entire scalp and are responsible for the growth of the hair. The follicles contain cells of proliferative epithelium origin at the base, or root, of the follicle.

[0003] In addition, the direct care of hair fibers also includes the treatment of their component proteins. For example, keratin is particularly susceptible to damage caused by urban lifestyles, frequent use of chemical hygiene products, hair-styling habits, and also environmental conditions, i.e., sun, pollution, salt, wind, fluctuations in climatic conditions, general lifestyle, and age. These proteins are denatured, resulting in skin appendages, especially hair, which are less durable, less flexible, stiff, dull, and prone to split ends. These appendages lose their biomechanical and cosmetic properties.

[0004] One solution is to supply exogenous proteins to compensate for the damage to the hair proteins.

[0005] However, hydrolysis (chemical, thermal, or enzymatic) is a necessary process to break down proteins to lower molecular weight, increase their water solubility, and in this case, to penetrate them into the hair fiber, making them usable in cosmetic formulations. Only low molecular weight molecules are able to penetrate into the fiber, especially damaged hair fibers. The hydrolyzed proteins help to improve moisture retention, elasticity of the hair fiber, and enhance softness, sheen, bounce, and body. They help to strengthen the hair fiber from the inside.

[0006] High-performance hydrolyzed proteins have substantial ability on hair fibers (weak binding with hair keratin) due to ionic charges and polar sites (interactions by van der Waals forces). They can also form a protective film on the surface of hair fibers.

[0007] Products based on hydrolyzed proteins exist in the cosmetics market. These proteins are of vegetable or animal origin. The most common are hydrolyzed proteins from wheat, silk, keratin, collagen, elastin, milk and almond. However, there is a constant demand for effective alternative ingredients in this field.

[0008] Quite unexpectedly, the inventors have found that the hydrolysate of H. rhamnoides cake has the capacity to penetrate keratin fibres and improve their mechanical properties, as well as their textural and superficial properties. It has been shown that this hydrolysate is able to increase the growth of skin appendages, combat their loss, increase their resistance and, in particular in the case of hair, reduce their breakage, making them shinier, brighter and more voluminous. In addition, the hydrolysate of H. rhamnoides cake also has the capacity to repair skin appendages, as will be shown below.

[0009] One advantage of the hydrolysate according to the invention is that it is a product derived from industrial extraction residues that are not usually upgraded, and therefore the development of such a product is included in the eco-responsible approach. Another advantage is that it is a complete care ingredient, since it is effective for the keratin fibres, in particular the hair, but also for the scalp and hair follicles. In addition, the hydrolysate according to the invention can be easily produced on an industrial scale. Finally, the enzymatic hydrolysis makes it possible to fractionate the proteins of the cake and recover the low molecular weight peptides, which are capable of penetrating the keratin fibres and providing the cosmetic effects described herein.

[0010] The hydrolysate according to the invention is obtained from the cake of the Hippophae rhamnoides plant. This plant, also called sea buckthorn, is known to have been used historically. Thus, in ancient Greece, its leaves were used as fodder for animals, especially horses, to increase the shine of their fur. The Greeks also used their fruits to therapeutically suppress pain and stomach pains, and to treat scurvy. Extracts of the leaves were also used to treat asthma, stomach ulcers, skin diseases (scars, burns), and lung diseases. In China, the fruits of H. rhamnoides are used to calm coughs, clear phlegm, strengthen the stomach, and to relieve indigestion, improve blood circulation, and dissipate congestion. In Central Asia, locals used sea buckthorn to treat high blood pressure, and digestive and skin diseases. The oil extracted from the fruits is used for its anti-inflammatory properties and to treat stomach ulcers, and a decoction of the dried fruits is used for skin disorders.

[0011] Moreover, extracts of H. rhamnoides are already present in the cosmetic market. Therefore, the supercritical CO 2 A water extract of the by-product from the extraction is sold by the applicant under the name RNAge™ for its anti-ageing properties on the skin and mucous membranes and is described in patent application WO 2019 / 069007. However, the extract in the present invention is a product of the supercritical CO2 extraction of the seeds. 2 It differs from the latter in that it is a hydrolysate of a by-product from the extraction. The cosmetic benefits described in this invention are further different.

[0012] Patent applications China Patent No. 105063139 and China Patent No. 108065411 describe the use of supercritical CO2 extracts of H. rhamnoides seeds. 2 Peptide hydrolysates from the residues of the extraction are disclosed. They are hydrolysates obtained by enzymatic hydrolysis. However, in neither of these two patent applications is there any disclosure of cosmetic or dermatological applications, which form the subject of the present invention.

[0013] Patent application CN108354862 discloses a composition containing, inter alia, oil of H. rhamnoides for anti-dandruff and soothing effects. However, firstly, it is an oil and not a hydrolysate, and secondly, its cosmetic effect described in the present invention is different from the anti-dandruff effect of said patent application.

[0014] The patent application FR 3031455 describes several molecules that can be extracted from H. rhamnoides and that are intended to be incorporated in cosmetic compositions, inter alia, as restorative agents for hair. However, there is no disclosure or suggestion of the exact effect as described in the present invention. Furthermore, an extract of the plant H. rhamnoides is disclosed, but without further explanation of the type of extract.

[0015] Patent application FR 2943255 describes the use of supercritical CO to promote the activity of α-5-reductase in order to improve hair shine loss. 2 The use of an extract of H. rhamnoides oil obtained by extraction with supercritical CO has been described. 2 It is an extract product, not a by-product thereof, much less a hydrolysate thereof.Furthermore, there is no disclosure or suggestion of any cosmetic benefits as described in this patent application.

[0016] The patent application WO 2009 / 125071 brochure states that supercritical CO 2 Several plant extracts are described, which are obtained from the by-products of the extraction, which are then subjected to an aqueous-alcoholic extraction, or in particular to an extraction with a mixture of water and butylene glycol. An extract of the fruits of H. rhamnoides is disclosed. However, the extract of the present invention is different in that it is a hydrolysate of a by-product of H. rhamnoides. The said patent application furthermore makes no mention of the cosmetic effect described in the present invention, and in particular of the effect on the skin appendages. Summary of the Invention [Problem to be solved by the invention]

[0017] Thus, to the best of the applicant's knowledge, the prior art does not describe the cosmetic and pharmaceutical use of the hydrolysates according to the invention, nor is said use suggested in any document, either alone or in combination. [Means for solving the problem]

[0018] Thus, a first subject matter relates to the pharmaceutical and cosmetic use of a hydrolysate of a cake of H. rhamnoides for increasing the growth of skin appendages and / or reducing their loss, advantageously of hair; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histostructural properties of skin appendages, advantageously of hair; and / or for repairing damaged skin appendages, advantageously of hair.

[0019] A second subject concerns the non-pharmaceutical cosmetic use of the hydrolysates thereof in cosmetic compositions.

[0020] A third subject relates to a non-medicinal cosmetic care process comprising the topical application of a hydrolysate according to the invention or a cosmetic composition containing it.

[0021] A fourth subject relates to a cosmetic treatment method and a last subject relates to the hydrolysate according to the invention for its dermatological or pharmaceutical use.

[0022] A first subject matter therefore relates to the non-pharmaceutical cosmetic use of a hydrolysate of H. rhamnoides cake for increasing the growth of skin appendages and / or reducing their loss, preferably of hair; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histostructural properties of skin appendages, preferably of hair; and / or for repairing damaged skin appendages, preferably of hair. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] The term "cosmetic use" refers to a non-therapeutic, non-pharmaceutical, non-dermatological use, i.e. a use not requiring a therapeutic treatment, and intended for healthy skin and / or skin appendages and / or mucous membranes. The term "healthy" refers to skin or skin appendages or mucous membranes that are determined by a specialist in the field, a dermatologist, to be non-pathological, i.e. to be free of any infection, inflammation, scarring, skin disease or skin condition, such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, in particular seborrheic dermatitis, dandruff, or wounds or injuries, and / or other skin diseases, and / or alopecia and / or baldness. Thus, for the purposes of the present invention, a "damaged" skin appendage is an appendage that is determined by a specialist in the field to be non-pathological. Specifically, damaged skin appendages are those that have lost their suppleness and / or elasticity and / or deformability, and in particular and as a consequence, their moisture. "Damaged" skin appendages have lost their structure and / or their visual and / or biomechanical properties. They therefore become dull and rough, with reduced durability, friable, brittle, split and / or curl, resulting in split ends. Furthermore, they become stiff, therefore losing softness and being difficult to style and shape. This loss of surface quality of the hair is visible and unsightly. The hair also has reduced light reflection, therefore visibly becoming dull and less shiny. The hair also becomes thinner and loses thickness.

[0024] The term "skin appendages" means herein the hair, eyelashes, eyebrows, body hair, in particular the beard, and / or nails. In the present invention, the beard also includes the moustache. It is preferred if the skin appendage concerned is the hair. The hydrolysate according to the invention is a topically acceptable ingredient. The term "topically acceptable" means an ingredient which is suitable for topical application, which is non-toxic, non-irritating, does not induce an allergic or inflammatory reaction on the skin, in particular on the scalp, or on the mucous membranes or skin appendages, and is not chemically unstable.

[0025] The hydrolysate can be used orally or topically. Advantageously, it is used topically. The term "topically" refers to local direct application and / or spraying the ingredient onto the skin and / or mucous membranes and / or skin appendages, especially onto the surface of the skin appendages. In a highly advantageous mode, the topical route excludes application onto the skin, including the scalp and / or mucous membranes.

[0026] The hydrolysate may be applied topically to all or part of the skin of the body and / or face selected from the scalp, legs, thighs, arms, upper abdomen, nape, neck, all or part of the face, forehead, chin, lips, lip outline, eye outline, "T" area of ​​the face, and advantageously to the scalp, and / or to all or part of the skin appendages, advantageously to the nails, head hair, body hair, in particular the beard, eyelashes and / or eyebrows, even more advantageously to the head hair, more preferably to the head hair.

[0027] The term "skin" also refers to the scalp.

[0028] The subject of the present invention therefore relates to the non-pharmaceutical cosmetic use of a hydrolysate of H. rhamnoides cake for increasing the growth of skin appendages and / or reducing their loss, advantageously of hair; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histostructural properties of skin appendages, advantageously of hair; and / or for repairing damaged skin appendages, advantageously of hair.

[0029] For the purposes of the present invention, the term "maintaining and / or increasing the biomechanical properties" of skin appendages means maintaining and / or increasing their resistance and / or elasticity and / or deformability and / or strength and / or plastic properties, i.e. their fragility, in particular their response to elongation and / or their flexibility. These properties can be evaluated ex-vivo by measuring their resistance to tensile forces. These biomechanical parameters, in particular those of hair, can be evaluated in response to elongation and can be measured, for example, by tensile tests (Dia-Stron). The parameters measured can be the modulus of elasticity (Pa), elongation at break (%), strength at break (gmf), gradient of the post-yield plastic region (gmf / % elongation), normalized in relation to the diameter of the skin appendage. This last parameter (slope) allows the measurement of the plasticity of the material studied just before breakage. The method makes it possible to obtain stress (i.e. force) curves as a function of the characteristic elongation of the material studied. It is known that in hair fibers (skin appendages) damaged by oxidation, reduction or UV irradiation, followed by cleavage in the cortical domain, resulting in the disulfide bridges becoming free radicals, an increase in the elongation at break and a decrease in the slope after the yield point are observed.

[0030] In one embodiment of the invention, "maintaining and / or increasing the biomechanical properties" of the skin appendages means reducing the production of Advanced Glycation End Products (AGEs), which are manifested in particular during chrono-induced or photoinduced aging, but also in response to environmental conditions, in contact with oxidative chemical or physical agents, and on a daily basis. These glycation products reduce the biomechanical properties of the appendage. Thus, in one embodiment, the hydrolysate according to the invention is in an amount effective to "maintain and / or increase the biomechanical properties of the skin appendage", such as when the percentage of glycation products measured after oxidation of albumin as marker protein in the presence of iron as catalyst and hydrogen peroxide as oxidizing agent and in the presence of the hydrolysate according to the invention is reduced by at least 20%, advantageously at least 25%, more advantageously at least 50%, based on the percentage of glycation products detected under the same conditions but without the extract. Advantageously, the skin appendage is hair. More advantageously, this is a reduction in the glycation products detected in the presence of the hydrolysate prepared according to Example 1a) and under the conditions described in Example 3a) (Table 3).

[0031] In an alternative embodiment of the invention, "maintaining and / or increasing the biomechanical properties" of the skin appendage means that, as a result of the oxidation of albumin in the presence of hydrogen peroxide and iron as oxidation catalyst, the decomposition of tryptophan measured in the presence of the hydrolysate according to the invention is reduced by at least 30%, advantageously at least 40%, more advantageously at least 95%, based on the percentage of decomposition of tryptophan measured in the absence of the hydrolysate according to the invention. Advantageously, the skin appendage is hair. More advantageously, it is a hydrolysate prepared according to example 1a) under the conditions described in example 3b) (table 4).

[0032] In another alternative embodiment of the invention, "maintaining and / or increasing the biomechanical properties of the skin appendage" means that the formation of dityrosine detected in the presence of iron and hydrogen peroxide after oxidation of albumin is reduced by at least 35%, preferably at least 55%, in the presence of a hydrolysate according to the invention. In an advantageous embodiment, the skin appendage is hair. More advantageously, the hydrolysate is the hydrolysate prepared according to example 1a) under the conditions described in example 3c) (table 5).

[0033] In yet another alternative embodiment of the invention, "maintaining and / or increasing the biomechanical properties of the skin appendage" means that the formation of pentosidine detected in the presence of iron after oxidation of albumin in the presence of hydrogen peroxide is reduced by at least 35%, preferably at least 60%, in the presence of a hydrolysate according to the invention. In an advantageous embodiment, the skin appendage is hair. More advantageously, the hydrolysate is the hydrolysate prepared according to Example 1a) under the conditions described in Example 3d) (Table 6).

[0034] In yet another embodiment of the invention, "maintaining and / or increasing the biomechanical properties of the skin appendage" means that in the presence of copper and lipoproteins, in the presence of a hydrolysate according to the invention, the production of Schiff bases, glycation intermediates, is reduced by at least 11%, advantageously by at least 18%, and very advantageously by at least 93%, relative to the amount of said Schiff bases measured under the same conditions but without the hydrolysate. In an advantageous embodiment, the skin appendage is hair. More advantageously, the hydrolysate is the hydrolysate prepared according to Example 1a) under the conditions described in Example 3d) (Table 7).

[0035] The hydrolysates according to the invention are therefore effective and can be used to increase the resistance, elasticity, flexibility, strength and / or plasticity of skin appendages, preferably those of hair, and reduce their breakage.

[0036] The term "maintaining and / or increasing the superficial and / or textural properties" of the skin appendage, advantageously the hair, further means making said skin appendage smoother and / or softer and / or less stiff and visually shinier and / or shinier, and therefore easier to disentangle and / or to style and / or to shape. These properties can be evaluated by imaging techniques, such as video or electron microscopy, or by sensory tests on hair tresses carried out by external volunteers trained to define visual or tactile sensations, or by expert (hairdresser) evaluation tests or consumer self-evaluation by questionnaire. The effectiveness of the product in its formulation is evaluated according to perceptible visual or tactile quality evaluation criteria of the hair.

[0037] In addition, the term "maintaining and / or increasing the growth of skin appendages" means maintaining and / or increasing the amount of DNA in the fibroblasts of the hair papilla. In one embodiment, this refers to an increase in the amount of DNA measured in the fibroblasts of the hair papilla in the presence of the hydrolysate according to the invention by at least 20%, preferably at least 30%, more preferably at least 40%, compared to the amount of DNA measured on non-sensitized fibroblasts without hydrolysate. Even more preferably, the hydrolysate according to the invention is as described in Example 1a) under the measurement conditions as shown in Example 4 (Table 8).

[0038] In an alternative embodiment of the invention, the term "maintaining and / or increasing the growth of skin appendages" means increasing the amount of ATP in the fibroblasts of the hair papilla. In one embodiment, this means that the amount of ATP measured in the fibroblasts of the hair papilla in the presence of a hydrolysate according to the invention is increased by at least 10%, preferably at least 15%, even more preferably at least 40% compared to the amount of ATP measured without hydrolysate. Even more advantageously, the hydrolysate according to the invention is that described in Example 1a) under the measurement conditions as shown in Example 4 (Table 9).

[0039] In yet another alternative embodiment of the invention, "maintaining and / or increasing the growth of the skin appendages" means maintaining and / or increasing the synthesis of fibroblasts in the papilla of the hair follicle of said appendages. In one embodiment of the invention, this means that the number of fibroblasts in the papilla in the presence of the hydrolysate according to the invention is increased by at least 10%, preferably at least 20%, more preferably at least 25% compared to the number of fibroblasts measured without said hydrolysate. Advantageously, it is an increase in the number of fibroblasts measured in the papilla of the hair follicle. More advantageously, this increase is measured in the presence of a hydrolysate prepared according to Example 1a) under the conditions described in Example 4 (Table 10).

[0040] It is also possible to evaluate ex vivo the effect of the hydrolysate on increasing the growth of skin appendages, in particular hair. Thus, in yet another alternative embodiment of the invention, the term "increasing the growth of skin appendages" means an elongation of keratin fibres in the presence of a hydrolysate according to the invention, advantageously in the presence of an agent that sensitises said fibres and reduces their elongation, of at least 5%, advantageously at least 10%, compared to the elongation of control keratin fibres sensitised with said agent without hydrolysate. Advantageously, this relates to hair. It is highly advantageous if the sensitiser is capsaicin. It is even more advantageous if the hydrolysate of H. rhamnoides cake is prepared according to the conditions described in Example 5 and as described in Example 1a).

[0041] In addition, the term "reducing the loss of skin appendages" means that the amount of DNA in the fibroblasts of the hair follicle papilla is increased when sensitized with the dedicated agent. Advantageously, therefore, the amount of DNA measured in the fibroblasts sensitized with capsaicin in the presence of the hydrolysate according to the invention is increased by at least 25%, advantageously at least 50%, and very advantageously at least 60% compared to the amount of DNA measured in the same fibroblasts sensitized with capsaicin without the hydrolysate. In a particularly advantageous embodiment of the invention, this includes an increase in the amount of DNA measured in the hair follicle papilla. More advantageously, this increase is measured in the presence of the hydrolysate prepared according to Example 1a) under the conditions described in Example 4 (Table 8).

[0042] In another mode of the invention, "reducing the loss of skin appendages" means that in the presence of capsaicin and the hydrolysate according to the invention, the number of papilla fibroblasts sensitized with capsaicin is increased by at least 20%, preferably at least 30%, more preferably at least 50%, compared to the number of fibroblasts sensitized with capsaicin without the hydrolysate. In a particularly advantageous embodiment of the invention, this is an increase in the number of fibroblasts measured in the papilla of the hair follicle. More advantageously, this increase is measured in the presence of the hydrolysate prepared according to Example 1a) under the conditions described in Example 4 (Table 10).

[0043] In yet another alternative embodiment of the invention, "reducing the loss of skin appendages" means that the amount of ATP in the fibroblasts of the papilla of the hair follicle, preferably when sensitized with capsaicin, is increased. In an advantageous embodiment, this means that the amount of ATP measured in the fibroblasts of the papilla, advantageously subjected to sensitization with capsaicin in the presence of a hydrolysate according to the invention, is increased by at least 100%, preferably at least 130%, when measured in the fibroblasts sensitized with capsaicin in the presence of a hydrolysate according to the invention, compared to the amount of ATP measured in the same fibroblasts sensitized with capsaicin without hydrolysate. In a particularly advantageous embodiment of the invention, this includes an increase in the amount of ATP measured in the papilla of the hair follicle. More advantageously, this increase is measured in the presence of a hydrolysate prepared according to Example 1a) under the conditions described in Example 4 (Table 9).

[0044] The properties of increasing the growth of the skin appendages, in particular the hair, and of increasing the biomechanical and / or superficial and / or textural properties of the hydrolysate according to the invention make it an active ingredient for improving the volume, color, sheen and radiance of the skin appendages, in particular the hair.

[0045] The effectiveness of the hydrolysates according to the invention on the biomechanical properties of the skin appendages, in particular of the hair, can also be demonstrated by various tests, such as tensile strength tests (Diastron), fatigue tests or repeated styling tests on hair, which allow the evaluation of breakage. It is also possible to quantify hair fibres with split ends. Furthermore, their biomechanical properties can also be evaluated by measuring the styling force required on dry or damp hair.

[0046] In addition, it is also possible to evaluate the penetration of the hydrolysate by directly measuring its distribution in the skin appendages. The distribution of the hydrolysate is advantageously studied on the hair by various imaging and microscopic methods. It is more advantageous if Fourier transform infrared spectroscopy (FTIR) can be used. Thus, in an advantageous embodiment of the invention, the hair is cut transversely after application of the hydrolysate according to the invention to hair damaged with an oxidizing agent, and then studied by FTIR to compare with the same hair damaged with the same oxidizing agent without application of the hydrolysate, or with undamaged hair. The vibration profile of the hydrolysate is then compared with the vibration profile of the section to evaluate the distribution of the product inside the hair. It is more advantageous if the oxidizing agent is hydrogen peroxide.

[0047] In addition, the term "repairing damaged skin appendages" refers to the repairing effect of the hydrolysate according to the invention on skin appendages damaged by: nutritional deficiencies; pollutants selected from fine particles known as PM2.5 and PM10, metals such as copper or iron, heavy metals; climatic and / or environmental conditions such as wind, rain, sudden changes in temperature and humidity, cold, heat, UV; sea salt, chlorine from swimming pools, and other physical or chemical reactive agents such as hydrogen peroxide, present in particular in certain shampoos, cosmetic make-up products, hair coloring, bleaching and perm products, varnishes, solvents and household products; mechanical attacks such as brushing; heat, for example from hair dryers and hair straighteners; and also natural and / or chrono-induced aging. The latter in fact also leads to a loss of structural and / or functional properties (including loss of moisture) of the skin appendages, in particular the eyelashes, eyebrows and hair, causing them to deteriorate.

[0048] For the purposes of the present invention, the term "repairing the skin appendage" also means at least partially improving the structural and / or visual and / or biomechanical properties of the skin appendage, preferably the hair, which is preferably assessed upon exposure to environmental, chemical, biological and / or mechanical stresses.

[0049] Environmental stresses include, inter alia: smoke, pollution, temperature, especially sudden fluctuations in temperature, hot or cold, moisture, especially humid and dry, sunlight, especially in the visible spectrum, UV, and / or gamma radiation, rain, wind, dust, and sea salt.

[0050] Chemical stresses include: harsh household products, chlorine from swimming pools, harsh cosmetic products such as coloring and bleaching products, nail polish and / or remover, solvents, and make-up.

[0051] In particular, chemical stresses on the hair include: aggressive shampoos and hair care and / or treatment products, in particular for styling and shaping, for example straightening and / or perming, and / or for dyeing and / or bleaching.

[0052] Mechanical stresses include in particular: friction, e.g. brushing and / or rubbing against fibres (ears, fabrics) and / or particulate matter (e.g. dust and / or sand), heat from hairdryers and hair straighteners, and / or hair styling, especially when exposed to pulling, stretching and / or twisting forces.

[0053] The hydrolysates according to the invention are furthermore effective as anti-aging ingredients for the skin appendages, preferably the hair, and / or the skin, preferably the scalp, and / or mucous membranes, by reducing the negative effects of aging.

[0054] Thus, in one embodiment of the present invention, the restorative effect can be evaluated according to existing techniques in the field.Using conventional methods for measuring the restorative effect, it is possible to measure the ability of the products to be evaluated to restore the visual, structural and / or functional state of damaged skin appendages, preferably damaged hair, in comparison with the state of undamaged skin appendages, preferably undamaged hair.They are applied to damaged skin appendages, preferably damaged hair, and the restorative effect is measured by comparing it with undamaged skin appendages, preferably undamaged hair.It is advantageous if the restorative effect is a restorative effect on damaged skin appendages in vitro using an oxidizing agent, which induces the denaturation of the proteins of the appendages.It is advantageous if it is a restorative effect of the hydrolysate on hair, and the oxidizing agent is more preferably hydrogen peroxide. More advantageously, the protein denaturation is assessed by measuring the protein denaturation temperature (°C) by differential scanning calorimetry (Wortmann and Deutz, Appl. Polym. Sci., 48, 137-150 (1993)) using a differential enthalpy analyzer (DSC Q100, TA Instruments) under the conditions described in Example 6. The hydrolysate according to the invention is thus present in an amount effective to repair skin appendages, preferably hair, such that the repair index of the damaged appendage is at least 20%, preferably at least 30%, compared to a "healthy" appendage, i.e. an appendage not subjected to hydrogen peroxide-induced oxidative stress. Advantageously, this includes measuring the repair index in the presence of a hydrolysate prepared according to Example 1a) under the conditions described in Example 6.

[0055] To assess the repair effect on skin appendages, preferably hair, in vivo, several methods can be used, selected from optical microscopy (video microscopy), confocal microscopy, FTIR or Raman microscopy, X-ray methods and electron microscopy, the purpose of which is to observe the state and / or quality of the cuticle, the protective covering of the keratin fibers of the skin appendage, by visualizing and quantifying detached scales as a sign of surface damage. To assess the morphology of the skin appendages, especially the hair, physical surface quantification methods can be carried out, such as atomic force microscopy or white light interference profilometry, for example, to assess the chemical (XPS), charge (streaming potential) or energy (inverse gas chromatography).

[0056] To evaluate the restorative effect of the hydrolysates according to the invention on the internal properties of the skin appendages, and in particular of the hair, calorimetry (DSC) can be used.

[0057] The hydrolysate according to the invention is made by subjecting the seeds of H. rhamnoides to supercritical CO 2It is a hydrolysate obtained by enzymatic or chemical hydrolysis of a by-product from the extraction, i.e. from the oil or from the by-product obtained after squeezing the seeds. For the purposes of the present invention, the seeds correspond to the berries (i.e. the fruit) of H. rhamnoides, excluding the pulpy part (also called the pericarp). Supercritical extraction with carbon dioxide is a technique familiar to the skilled person, which allows the isolation of an oily fraction of the compounds extracted, which can then be used in various applications. For the purposes of the present invention, the residue of this extraction is called "coproduct", which includes all the compounds that were not extracted via said technique. The term "coproduct" therefore means the residue (also called cake) obtained after the extraction of the oil. In the following description, the term "cake" will be used. It is therefore advantageously a defatted fraction of the seeds of H. rhamnoides. That is, supercritical CO2 of H. rhamnoides seeds 2 Preferably it is an enzymatic or chemical hydrolysate of a by-product from the extraction, more preferably an enzymatic hydrolysate, more preferably the by-product does not contain any oil.

[0058] The enzymatic hydrolysis can be carried out in the presence of various proteases known to those skilled in the art, preferably in the presence of: enzymes of animal origin selected from pepsin; enzymes of pancreatic origin, such as trypsin or chymotrypsin, preferably trypsin; enzymes of plant origin selected from papain, bromelain, ficin or actinidin, preferably papain; or enzymes of bacterial origin selected from the Bacillus licheniformis system (sold under the name Alcalase®) or from the system B. subtilis, preferably Alcalase®. In a particularly advantageous embodiment of the invention, the enzyme used is Alcalase®.

[0059] The hydrolysis is preferably carried out at a pH between 3 and 9, advantageously between 4 and 8.5, and very advantageously at a pH of 8.5, depending on the optimum pH of the enzyme. The hydrolysis is preferably carried out at a temperature between 40° C. and 65° C., advantageously between 50 and 60° C., and very advantageously at 55° C. It is carried out for a period of 1 to 3 hours, preferably for 2 hours.

[0060] The enzyme used is then inactivated by heating for a period of 5 to 30 minutes, preferably 10 minutes, advantageously at a temperature between 80° C. and 100° C., very advantageously at a temperature of 90° C. Enzyme inactivation occurs at a pH between 5 and 8, preferably at pH 6.5.

[0061] The hydrolysate so obtained is then centrifuged and purified by successive filtration to a porosity of 0.22 μm.

[0062] Prior to the enzymatic hydrolysis step, the proteins are advantageously first extracted from the cake, in which case they are extracted at a pH between 7.5 and 10, advantageously between 8 and 9, and very advantageously at a pH of 9.0, for a period of 30 minutes to 2 hours, preferably 1 hour.

[0063] The weight of the cake used for protein extraction and hydrolysis is between 5% and 20%, preferably between 5% and 15%, more preferably it is 10% by weight, based on the total weight of the solvent and the cake.

[0064] The solvent used for protein extraction is chosen from the following: water, coconut water as solvent as described in patent application FR 3 061 416, or at least one C 2 O 4 as described in patent application FR 3 069 450, based on the total weight of the solvent. 6 ~C 16 Dialkyl carbonates, e.g. C 7 ~C 10 , advantageously C8 A solvent comprising at least 50% by weight of a dialkyl carbonate, such as dioctyl carbonate or diethylhexyl carbonate. Advantageously, the solvent used is water as the sole solvent.

[0065] The protein extraction is preferably carried out at a temperature between 4° C. and 300° C., advantageously between 4° C. and 100° C., more advantageously between 15° C. and 80° C., and most advantageously between 15° C. and 30° C. (including room temperature, i.e. a temperature of 20° C.). In a particularly advantageous embodiment of the invention, the protein extraction is carried out at room temperature.

[0066] In an alternative embodiment, the extraction is carried out under subcritical conditions.

[0067] The term "extraction under subcritical conditions" refers to extraction in the presence of water, at temperatures above 100°C, and at pressures below 221 bar, where the water still remains in a liquid state but has a lower viscosity and surface tension than water at room temperature and a higher dielectric constant. The extraction pressure will therefore be between 100 bar and 250 bar, preferably between 150 and 221 bar.

[0068] Thus, under subcritical conditions, the extraction is carried out in water at a temperature ranging between 100°C and 300°C, advantageously between 120°C and 250°C, more advantageously between 140°C and 200°C. The extraction may be carried out at a single pre-determined temperature or at a successively increasing temperature. In one advantageous embodiment of the invention, the extraction will be carried out at a single temperature of 160°C. In an alternative embodiment, it will be carried out following a three-stage temperature gradient between 100°C and 200°C, such as 120°C, 140°C, then 160°C; or 110°C, 130°C, then 150°C; or else 120°C, 145°C, then 170°C.

[0069] In a particularly advantageous embodiment of the invention, the hydrolysate is obtained by enzymatic digestion as follows: the protein is extracted from the cake in an amount of 10% by weight, based on the total weight of the cake and water as solvent, at room temperature and pH 12 for 1 hour. The protein thus extracted is hydrolyzed in the presence of Alcalase® at a concentration of 5% by volume, based on the total volume of Alcalase® and protein, at a temperature of 55° C. and pH 8.5 for 2 hours. The enzyme is inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture is then cooled, centrifuged and filtered (0.22 μm) under the conditions described in Example 1a).

[0070] Alternatively, the proteins of the cake are extracted at room temperature, i.e. at a temperature of 20° C. and pH 8.5 for 1 hour, using 10% by weight of the cake, based on the total weight of the cake and water as solvent. The proteins thus extracted are hydrolyzed in the presence of Alcalase® at a concentration of 5% by volume, based on the total volume of Alcalase® and protein, at a temperature of 55° C. and pH 8.5 for 2 hours. The enzyme is inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture is then cooled, centrifuged and filtered (0.22 μm) under the conditions described in Example 1b).

[0071] Separately, the protein of the cake is extracted at room temperature, i.e., at a temperature of 20° C. and pH 12 for 1 hour, using 10% by weight of the cake, based on the total weight of the cake and water as solvent. The protein thus extracted is hydrolyzed in the presence of papain, at a concentration of 5% by volume, based on the total volume of papain and protein, at a temperature of 55° C. and pH 8.5 for 2 hours or more. The papain is inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture is then cooled, centrifuged and filtered (0.22 μm) under the conditions described in Example 1c).

[0072] The hydrolysate according to the invention therefore comprises: a dry matter content of 1% to 20% by weight, advantageously 2% to 10% by weight, for example 5%; a total protein content of the hydrolysate of 15 g / L to 35 g / L, advantageously 18.8 g / L; furthermore a percentage of peptides with a molecular weight of 5 kDalton (Da) to 30 kDa between 15% and 40%, advantageously 18.25%; a percentage of peptides with a molecular weight of less than 5 kDa between 40% and 85%, advantageously 72.6% (Example 2). Advantageously, the hydrolysate comprises a dry matter content of at least 3% by weight, in particular at least 3.5% by weight, more particularly 3.44% by weight, very advantageously 3.84% by weight. In an even more advantageous mode, the hydrolysate comprises a dry matter content of 5% by weight.

[0073] Very advantageously, the hydrolysate contains a percentage of peptides with a molecular weight between 5 kDa and 30 kDa of 18.25% and a percentage of peptides with a molecular weight below 5 kDa of 72.6%.

[0074] Very advantageously, the hydrolysate according to the invention further does not contain gallic acid or any polymer that can be hydrolyzed to gallic acid, or hexahydroxydiphenic acid or any polymer that can be hydrolyzed to hexahydroxydiphenic acid, or ellagic acid or any polymer that can be hydrolyzed to ellagic acid, or gallotannins or ellagitannins.More advantageously, the hydrolysate further does not contain any derivatives of the compounds mentioned, or any of their salts, in particular as described in patent application FR 031455.

[0075] In another advantageous embodiment, the hydrolysate according to the invention does not contain L-quebrachitol or other methyl inositols and / or inositols as described in WO 2009 / 125071.In particular, the hydrolysate according to the invention does not contain myricetin, quercetin, kaempferol or isorhamnetin.

[0076] The hydrolysates, especially those prepared in Examples 1a) to 1c), are therefore in liquid form. Optionally, the hydrolysates may then be dried, for example by freeze-drying or by spraying in the presence of maltodextrin. The hydrolysates are then in powder form.

[0077] In this case, the hydrolysates according to the invention, in particular obtained under the conditions described in Examples 1a) to 1c), are finely ground in the presence of maltodextrin in a concentration by weight between 20% and 90%, preferably between 40% and 80%, more preferably between 70% and 80%, based on the total weight of the powder obtained.

[0078] The hydrolysate according to the invention may be used alone or may be included in a cosmetic composition.

[0079] When it is used alone, in the form of a cosmetic or dermatological ingredient, it is preferably dissolved in an aqueous solution containing glycerol and is advantageously present in a concentration of 60% to 90%, more advantageously 70% to 85%, very advantageously 80% by weight, based on the total weight of the aqueous solution containing its hydrolysate.

[0080] In an alternative embodiment of the invention, the hydrolysate will be dissolved and / or diluted in a solvent, in particular a polar solvent, such as water, alcohol, polyol, glycol, such as pentylene glycol and / or butylene glycol and / or propylene glycol and / or hexylene glycol and / or caprylyl glycol, or mixtures thereof, preferably water-glycol mixtures, more preferably those containing glycols selected from hexylene glycol, propylene glycol, caprylyl glycol, and various mixtures thereof. Advantageously, the hydrolysate thus obtained will be diluted and / or dissolved in an aqueous solution containing hexylene glycol, in particular between 0.1% and 10% by weight of hexylene glycol, preferably between 0.5% and 5% by weight of hexylene glycol, based on the total weight of the cosmetic ingredient. Advantageously, the hydrolysate thus obtained is diluted and / or dissolved in an aqueous solution containing caprylyl glycol, in particular between 0.01% and 5% by weight of caprylyl glycol, preferably between 0.1% and 1% by weight of caprylyl glycol, based on the total weight of the aqueous solution containing the hydrolysate.Alternatively, the solution in which the hydrolysate according to the invention is dissolved contains propylene glycol and caprylyl glycol.

[0081] Specifically, the aqueous solution in which the hydrolysate according to the invention is dissolved contains xanthan gum, particularly between 0.01% and 5% by weight xanthan gum, based on the total weight of the aqueous solution, and more particularly between 0.1% and 1% by weight xanthan gum, based on the total weight of the aqueous solution containing the hydrolysate.

[0082] Advantageously, the solution in which the hydrolysate according to the invention is dissolved comprises hexylene glycol, caprylyl glycol and xanthan gum.

[0083] In an alternative embodiment of the invention, the extract is dissolved in a solution containing a mixture of sodium benzoate and gluconolactone (sold under the name Geogard™).

[0084] The hydrolysate can be incorporated into a cosmetic composition comprising at least one cosmetically acceptable excipient. For the purposes of the present invention, the term "cosmetically acceptable" excipient means topically acceptable compounds and / or solvents, i.e., those that do not provoke adverse inflammatory and allergic reactions, are non-toxic, and are not unstable when in contact with the skin, particularly the scalp, or equivalents thereof.

[0085] For the purposes of the present invention, the term "cosmetic composition" means a non-pharmaceutical composition, i.e. a composition intended for protecting and / or caring for the skin, in particular the scalp, and / or skin appendages, which are considered "normal" by dermatologists, i.e. non-pathological. The term "normal" skin or scalp or skin appendages is intended herein to mean healthy skin or scalp or skin appendages, as defined above.

[0086] In a preferred embodiment of the invention, the hydrolysate according to the invention is incorporated into the cosmetic composition in an amount of 1×10 -4 % by weight to 10% by weight, preferably 1×10 -4 % by weight to 5% by weight, more preferably 1×10 -3 It is present in a content between 0.001% and 0.1% by weight.

[0087] The composition can therefore be used to increase the growth of skin appendages and / or reduce loss thereof, preferably of hair; and / or to maintain and / or increase the biomechanical and / or superficial and / or histostructural properties of skin appendages, preferably of hair; and / or to repair damaged skin appendages, preferably of hair.

[0088] The cosmetic compositions according to the invention may be in the form of a presentation customarily used for topical application to the skin or scalp and / or skin appendages, preferably to the scalp and / or skin appendages, such as in the form of a liquid or solid, or even in the form of a pressurized liquid. They may be formulated in the form of an aqueous or oily solution, a cream or an aqueous or oily gel (especially in a jar or tube), in particular a shower gel, a shampoo, a conditioner, a milky liquid, an oil, an emulsion, a hydrogel, a microemulsion or a nanoemulsion, in particular an oil-in-water or water-in-oil or hybrid or silicone-based emulsion, a serum, a lotion (especially in a glass or plastic bottle or a measuring bottle), or an aerosol or spray, a vial, a liquid soap, a paste, a dermatological bar, an ointment, a foam, a mask, a lacquer, a patch, a varnish, an anhydrous product (which is preferably in the form of a liquid, a paste or a solid, for example in the form of a wand, in particular in the form of a stick, or in the form of a powder). It may be a make-up product, in particular for the eyelashes or eyebrows, for example a mascara or pencil, or a make-up remover product. In particular, the cosmetic composition is selected from the group consisting of: a serum, a lotion, a cream, a shampoo, a hair conditioner, an oil, a milk, an ointment, a paste, a foam, an emulsion, a hydrogel, a shower gel, a mask, a lacquer, a spray, a wax, a mascara, a make-up pencil or a varnish, advantageously in the form of a shampoo, a conditioner or a lotion.

[0089] Preferably, the hydrolysate is suitable for the formulation of "neutral and mild" compositions intended for keratin fibers, especially hair fibers, and for the skin, especially the scalp. The hydrolysate is also suitable for use in cationic formulations with surfactants.

[0090] The compositions according to the invention may contain any suitable solvent and / or any suitable vehicle and / or any suitable additive, possibly in combination with other compounds of interest. These include in particular cosmetically acceptable additives selected from the following: surfactants, preservatives, buffers, swelling agents, chelating agents, biocides, denaturants, opacifiers, pH regulators, reducing agents, stabilizers, emulsifiers, thickeners, gelling agents, film-forming polymers, solvents, extenders, germicides, odor absorbers, matting agents, conditioning agents, texturizing agents, gloss agents, pigments, colorants, fragrances, chemical or mineral sunscreens, trace elements, and essential oils. Combinations thereof are also within the scope of the present invention. The CTFA Cosmetic Ingredient Handbook, Second Edition (1992) describes various cosmetic ingredients commonly used in the cosmetics industry, which are particularly suitable for topical application on the scalp.

[0091] The cosmetic composition may contain other cosmetic agents having similar properties to the hydrolysate according to the invention and exerting a synergistic or non-synergistic effect with said hydrolysate or may contain cosmetic agents having a complementary effect. Anti-hair loss active agents include combinations of sulfopeptides, amino acids, aminosaccharides, B vitamins, zinc and extracts of Panax ginseng and Artium majus (sold by the Applicant under the name Trichogen® LS8960), or protective agents for hair, such as extracts of the peel of Litchi chinensis (sold by the Applicant under the name Litchiderm®), or soothing and anti-itching active agents, such as rapeseed phytosterols (sold by the Applicant under the name Phytosoothe® LS9766).

[0092] Other active agents may also be present in the composition, such as, for example, an extract of Cassia alata leaves (sold under the name DN-Age™ as an antioxidant active agent, in particular for hair care), an extract of Salvia miltiorhizza in combination with niacinamide (sold under the name CollRepair™ as a deglycating agent), or active agents promoting firmness of the skin and thus of the scalp, such as synthetic tetrapeptides (sold under the name Dermican™), extract of Hibiscus abelmoschus (sold under the name Linefactor™), purified bean extracts (sold under the name Proteasyl™), extract of Manilkara multinervis (sold under the name Elestan™), extract of Khaya senegalensis ... Salvia miltiorhizza in combination with niacinamide (sold under the name CollRepair™ as a deglycating agent), or active agents promoting firmness of the skin and thus of the scalp, such as, for example, synthetic tetrapeptides (sold under the name Dermican™), extract of Hibiscus abelmoschus (sold under the name Linefactor™), extract of purified bean (sold under the name Proteasyl™), extract of Manilkara multinervis (sold under the name Elestan™), extract of Khaya senegalensis (sold under the name Proteasyl™), extract of Salvia miltiorhizza in combination with niacinamide (sold under the name CollRepair™), senegalensis extract (sold under the name Collalift 18), Argan pulp extract (sold by the Applicant under the name Argassential), Schizandra chinensis extract (sold under the name Sqisandryl), Eperua falcata extract (sold under the name Eperuline) or Orthosiphon staminus extract (sold by the Applicant under the name MAT-XS Bright). The combination of these active agents is particularly capable of strengthening hair follicles and reducing hair loss. The hydrolysate of the invention can also be combined with an extract of the seeds of the plant Nephelium lappaceum (sold by the Applicant under the name Rambuvital, in particular for its hair protection properties against pollution).

[0093] A third subject of the present invention relates to a non-pharmaceutical cosmetic care process for increasing the growth of skin appendages and / or reducing their loss, advantageously of hair; for maintaining and / or increasing the biomechanical and / or superficial and / or histostructural properties of skin appendages, advantageously of hair; and / or for repairing damaged skin appendages, advantageously of hair, comprising the topical application of a hydrolysate according to the invention or a cosmetic composition comprising it.

[0094] In a preferred embodiment of the invention, the cosmetic care process consists of the topical application of the hydrolysate according to the invention or of a cosmetic composition comprising it to: part or all of the skin of the body and / or face (selected from the scalp, legs, thighs, arms, midriff, nape, neck); part or all of the face, the forehead, the chin, the lips, the lip outline, the eye outline, the "T" area of ​​the face and preferably the scalp; and / or part or all of the skin appendages, preferably the nails, the hair, the body hair, in particular the beard, the eyelashes and / or the eyebrows, even more preferably the hair of the head, more preferably the hair of the head.

[0095] The cosmetic care process thus makes it possible to improve the colour and / or shine and / or radiance and / or volume of the skin appendages, advantageously of the hair.

[0096] Yet another subject matter relates to a cosmetic treatment method for increasing the growth of skin appendages and / or reducing the loss thereof, preferably of hair; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histoarchitectural properties of skin appendages, preferably of hair; and / or for repairing damaged skin appendages, preferably of hair, and comprising the following steps: - identifying a particular population not suffering from a pathology requiring therapeutic treatment, in particular a dermatological pathology, and / or a person in need thereof, to whom the application of a hydrolysate of H. rhamnoides or a cosmetic composition comprising it is desirable; - identifying healthy body areas and / or healthy skin (including healthy scalp and / or healthy skin appendages) in need of treatment; of the hydrolysate according to the invention or a composition containing it, in this case based on the total weight of the composition, -4 % by weight to 10% by weight, preferably 1×10 -4 % by weight to 5% by weight, more preferably 1×10 -3 topically applying at a content between 0.001% and 0.1% by weight and even more preferably between 0.001% and 0.1% by weight.

[0097] A final subject matter relates to a hydrolysate of the cake of H. rhamnoides for use alone or in a dermatological or pharmaceutical composition comprising at least one dermatologically or pharma- ceutical acceptable excipient, in the treatment of alopecia and / or alopecia. In an advantageous embodiment of the invention, the hydrolysate is that described in the present invention and is present in a dermatological or pharmaceutical composition in an amount of 1 × 10 6, based on the total weight of the composition. -4 % by weight to 10% by weight, preferably 1×10 -4 % by weight to 5% by weight, more preferably 1×10 -3 It is present in a content between 0.001% and 0.1% by weight and 3% by weight, more preferably between 0.001% and 0.1% by weight.

[0098] The examples form an integral part of the present invention, and any features believed to be novel over the prior art from the description taken in their entirety, including the examples, form an integral part of the present invention. Each example therefore has a general scope.

[0099] Unless otherwise specified, temperatures are expressed in °C and pressures are atmospheric. EXAMPLES

[0100] Example 1: Various methods for preparing cake hydrolysates Example 1a): The cake protein was extracted at room temperature, i.e. at a temperature of 20° C. and pH 12, for 1 hour using an amount of cake of 10% by weight based on the total weight of cake and water as solvent. The protein thus extracted was hydrolyzed in liquid form at a temperature of 55° C. and pH 8.5 for 2 hours using a volume concentration of 5% of Alcalase based on the total volume of Alcalase and protein. The enzyme was inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture was then cooled, centrifuged and filtered (0.22 μm).

[0101] Example 1b): The cake protein was extracted at room temperature, i.e. at a temperature of 20° C. and pH 8.5, for 1 hour using 10% by weight of the cake based on the total weight of the cake and water as solvent. The protein thus extracted was hydrolyzed in liquid form at a temperature of 55° C. and pH 8.5 for 2 hours using a volume concentration of 5% of Alcalase based on the total volume of Alcalase and protein. The enzyme was inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture was then cooled, centrifuged and filtered (0.22 μm).

[0102] Example 1c): The cake proteins were extracted at room temperature, i.e., at a temperature of 20° C. and pH 12, for 1 hour using 10% by weight of the cake based on the total weight of the cake and water as solvent. The proteins thus extracted were hydrolyzed at a temperature of 55° C. and pH 8.5 for 2 hours using a volume concentration of 5% of papain in liquid form based on the total volume of papain and protein. The enzyme was inactivated by heating at a temperature of 90° C. and pH 6.5 for 20 minutes. The mixture was then cooled, centrifuged and filtered (0.22 μm).

[0103] Example 2: Analysis of the total protein content and molecular weight profile of peptides of hydrolysates according to the invention Example 2a) Total Protein Assay: Materials and Methods: The protein content of the raw material and extracts was estimated by measuring total nitrogen (Kjeldahl method) and multiplying the value so obtained by 6.5 (N x 6.25).

[0104] result: The results are listed in Table 1 (Protein content of H. rhamnoides hydrolysates).

[0105] [Table 1]

[0106] Conclusion: The total protein content in the hydrolysates according to example 1a) is between 18 and 32 g protein / L (Table 1). The total protein content is between 37% and 65% on dry matter.

[0107] Example 2b) Analysis of the molecular weight profile of peptides: Materials and Methods: The molecular weight distribution of peptides of the hydrolysate prepared according to Example 1a) was analyzed by gel permeation on a column (Superose® 12 10 / 300 GL, GE Healthcare Life Sciences). Their molecular weights were determined after the column was calibrated with protein molecules of known molecular weight.

[0108] result: The results are listed in Table 2.

[0109] [Table 2]

[0110] Conclusion: The hydrolysate prepared according to example 1a) contains more than 70% of peptides with a MW below 5 kDa.

[0111] Example 3: In vitro effect of the hydrolysate according to the invention on the biomechanical properties of skin appendages Materials and Methods: 1.5% albumin, 2 mM iron, 5 mM EDTA, and 25 mM H 2 O 2 Reactants containing hydrogen peroxide were placed in the presence or absence of H. rhamnoides hydrolysate. 2 O 2 Albumin is oxidized with . The catalyst of the oxidation reaction is iron. The catalysis of the oxidation reaction is monitored by disappearance of iron. The mixture is incubated at 37 ° C for 1 day. The amount of tryptophan, dityrosine, pentosidine and glycation reaction products is measured by fluorescence (excitation / emission wavelength: 280 nm / 340 nm for tryptophan; 315 nm / 410 nm for dityrosine; 335 nm / 385 nm for pentosidine; 370 nm / 440 nm for general glycation reaction products).

[0112] result: The results are listed in Tables 3-6.

[0113] Example 3a): Reduction of glycation reaction products (AGEs)

[0114] [Table 3]

[0115] Conclusion: The presence of iron catalyzes the oxidation process of albumin in the presence of hydrogen peroxide, accompanied by the formation of fluorescent glycation products. The presence of H. rhamnoides hydrolysate at 0.01% and 0.1% (w / v) reduced the formation of these glycation products by at least 28%.

[0116] Example 3b): Reduction of tryptophan degradation

[0117] [Table 4]

[0118] Conclusion: The addition of the hydrolysate according to the invention reduces the degradation of tryptophan following the oxidation of albumin by at least 30%, indicating its ability to inhibit protein oxidation, so that the mechanical properties of the skin appendages, in particular the hair, are maintained and / or enhanced.

[0119] Example 3c): Reduction of the production of dityrosine

[0120] [Table 5]

[0121] Conclusion: The addition of the hydrolysate according to the invention reduces the formation of dityrosine after the oxidation of albumin by at least 35%, indicating that it has the ability to inhibit protein oxidation, so that the mechanical properties of the skin appendages, in particular the hair, are maintained and / or enhanced.

[0122] Example 3d): Reduction of Pentosidine Production

[0123] [Table 6]

[0124] Conclusion: The addition of the hydrolysate according to the invention reduces the formation of dityrosine after the oxidation of albumin by at least 35%, indicating that it has the ability to inhibit protein oxidation, so that the mechanical properties of the skin appendages, in particular the hair, are maintained and / or enhanced.

[0125] Example 3e): Reduction of Glycation Intermediates: Schiff Bases Materials and Methods: The hydrolysates prepared according to Example 1a) (final 0.01% or 0.1% (w / v)) were added (no addition in the control) to a buffer solution (phosphate buffered saline (PBS)) containing copper (200 μM) and lipoproteins (100 μg / mL). The mixture was incubated for 48 hours at a temperature of 37° C. The amount of Schiff base (average %) was measured spectrophotometrically (excitation wavelength 370 nm / emission wavelength 440 nm).

[0126] result:

[0127] [Table 7]

[0128] Conclusion: The hydrolysate according to the invention reduced the appearance of Schiff bases by at least 11%.

[0129] Example 4: Increased growth of skin appendages Materials and Methods: A suspension of fibroblasts from human hair follicle papillae and basal medium containing 0.05% v / v growth factor cocktail was centrifuged at 200 × g for 5 min with or without the hydrolysate of H. rhamnoides to be tested to form aggregates. Control conditions were constituted by adding 0.05% v / v growth factor cocktail (ethanol EtOH control) and capsaicin (10 μM) as a cell sensitizer (capsaicin control) to the basal medium. The aggregates were incubated in a regulated atmosphere (5% CO 2 The plates were incubated at 37° C. under a constant temperature of 100° C. and 95% relative humidity.

[0130] The aggregates were rinsed in buffered saline solution (PBS) and the cells were dissociated by incubation in a mixture of proteases (collagenase A, trypsin) and EDTA.

[0131] A portion of the suspended cells was taken up in PBS for flow cytometry analysis. Cell count and autofluorescence were the two parameters measured. Autofluorescence is a direct parameter that is easily measured and is used as a marker of cell senescence (Rattan et al., 1982). Cell count is an indirect parameter of hair follicle viability. Another portion of cells was used to analyze the cellular DNA content and ATP production rate. These two parameters were measured according to the supplier's instructions (CyQuant NF Cell proliferation Assay Kit C35006, Invitrogen and Bioluminescence Assay Kit CLS II Roceh 11699695001, Sigma-Aldrich).

[0132] result: The results are listed in the following tables: Table 8 (Effect of H. rhamnoides hydrolysate on the amount of DNA in papillary fibroblasts grown as aggregates), Table 9 (Effect of H. rhamnoides hydrolysate on ATP production by papillary fibroblasts grown as aggregates), and Table 10 (Effect of H. rhamnoides hydrolysate on the number of papillary fibroblasts grown as aggregates, their granularity, and autofluorescence measured by flow cytometry).

[0133] [Table 8]

[0134] Conclusion: Treatment of sensitized papillary fibroblast aggregates with capsaicin at a concentration of 10 μM reduces the amount of DNA in the cells. Co-addition of H. rhamnoides hydrolysate at 0.03% (w / v) reduces the negative effect of capsaicin, demonstrating the ability of the hydrolysate to reduce the loss of skin appendages, especially hair. Treatment of the aggregates with H. rhamnoides hydrolysate at 0.1% (w / v) increases the total amount of DNA (+41% compared to the control), demonstrating the efficacy of the hydrolysate according to the invention in increasing the growth of skin appendages.

[0135] [Table 9]

[0136] Conclusion: Treatment of sensitized papillary fibroblast aggregates with capsaicin at a concentration of 10 μM reduced the production of ATP in the cells by 76% compared to the control (ethanol). The combined addition of capsaicin and a hydrolysate of H. rhamnoides at 0.03% (w / v) made it possible: 1) to counteract the negative effects of capsaicin, demonstrating the ability of the hydrolysate according to the invention to reduce the loss of skin appendages, advantageously hair, and 2) to increase the production of ATP by at least 16% compared to the non-sensitized control, demonstrating the effect of said hydrolysate in increasing the growth of skin appendages, preferably hair.

[0137] [Table 10]

[0138] Conclusion: Treatment of sensitized papilla fibroblast aggregates with capsaicin (10 μM) reduced the number of papilla fibroblasts, increased the overall granularity of the cells, and increased the fluorescence, including hair loss, compared to the control (EtOH). The addition of H. rhamnoides hydrolysate at 0.03% (w / v) in combination with capsaicin reduced the negative effect of capsaicin on the three measured parameters and increased the cell count by 54% compared to the control, indicating that the hydrolysate inhibits hair loss. In addition, the number of cells measured in the presence of the hydrolysate was also higher than that of the control not sensitized with capsaicin, indicating its effect of increasing the number of fibroblasts and therefore on hair growth. These results indicated the protective potential of H. rhamnoides hydrolysate against premature or induced hair loss and its potential as a hair growth stimulant.

[0139] Example 5: Ex vivo effects on growth Materials and Methods: Human follicles were cultured ex vivo in complete growth medium containing 0.5 μg / mL insulin and treated with 30 μM capsaicin. The latter molecule is used as the agent responsible for retarding follicular growth. A control condition was also created without capsaicin treatment. Capsaicin-sensitized follicles were treated with or without 0.03% (w / v) H. rhamnoides hydrolysate. At least 10 isolated cultured follicles were evaluated for each culture condition (n=10). The follicles were incubated at 37°C, 5% CO 2 The cultured follicles were cultured in the presence of 0.1% ethanol. The medium was refreshed three times a week for approximately 2 weeks. Hair shaft elongation of the cultured follicles was obtained from photographs taken on different days. The hair elongation was assessed by image analysis (Image J) and expressed in μm as the mean ± SEM.

[0140] result: The results are listed in Table 11 (hair growth values, μm).

[0141] [Table 11]

[0142] Conclusion: Hair follicles cultured ex vivo in medium containing 0.5 μg / mL insulin continued to grow as evidenced by hair shaft elongation (control). Treatment with capsaicin (30 μM) caused a significant loss in elongation compared to controls by day 6 of culture. Treatment of hair follicles with 0.03% (w / v) H. rhamnoides hydrolysate sensitized with capsaicin significantly improved hair elongation as early as day 6 of culture compared to capsaicin-treated follicles.

[0143] Example 6: Repair effects on hair fibre Materials and Methods: Dark brown Caucasian hair locks were prepared for the study by trimming (1 g; 12 cm). The locks were washed and then soaked in a hydrogen peroxide solution (5.6% H 2 O 2 +13.9%(NH 4 ) 2 S 2 O 8 The hair tresses were bleached three times for 30 min using 100% ethyl acetate (pH = 9.4) at 20 °C. Healthy (unbleached) hair tresses served as controls. After washing and drying for 45 min under air flow at 55 °C, the tresses were immersed for 24 h in distilled water (control) or in an aqueous solution containing 1% H. rhamnoides hydrolysate. Both solutions were pre-buffered to pH 5.5. The tresses were rinsed and then dried for 1 h at approximately 60 °C under air flow. The denaturation temperatures of human proteins were determined by differential calorimetry using a heating rate of 2 K / min and a differential enthalpy analyzer (DSC Q100, TA Instruments) (Wortmann et al., 1993).

[0144] result:

[0145] [Table 12]

[0146] Conclusion: Hair bleached with hydrogen peroxide was less stable than healthy unbleached hair. The difference in denaturation temperature of hair fiber proteins was −8°C between damaged and healthy hair. The difference in denaturation temperature between peroxidized hair treated with H. rhamnoides hydrolysate and control hair was only −4.9°C. Treatment of damaged hair improved the stability of hair proteins against heat denaturation by +3.5°C. H. rhamnoides hydrolysate showed a restorative effect on hair fibers damaged by hydrogen peroxide.

[0147] Example 7: Examples of compositions containing hydrolysates according to the invention Example 7a): Shampoo

[0148] [Table 13]

[0149] The shampoo is prepared by mixing the four phases and adjusting the composition to a pH of 5.2 and a viscosity of 2200 mPa·s (measured using a Brookfield instrument (RVT; 23° C., spindle 5; 50 rpm)) by methods conventional in the art and well known to those skilled in the art.

[0150] Example 7b: Water-alcohol solution for the scalp

[0151] [Table 14]

[0152] The aqueous-alcoholic solution is prepared by mixing the three phases and adjusting the composition to a pH of 6.2 by methods commonly used in the art and known to those skilled in the art.

[0153] Example 7c: Mask for the scalp

[0154] [Table 15]

[0155] The mask is prepared by conventional methods known to those skilled in the art by mixing the four phases and adjusting the composition to a pH of 4.1 and a viscosity of 26,000 mPa·s (measured using a Brookfield instrument (RVT; 23° C., spindle 5; 50 rpm)).

[0156] Example 7d: Hair serum

[0157] [Table 16]

Claims

1. A cosmetic composition for increasing the growth and / or reducing loss of skin appendages; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histoarchitectural properties of skin appendages; and / or for repairing damaged skin appendages, the composition comprising a hydrolysate of a cake of Hippophae rhamnoides.

2. The composition described in claim 1, characterized in that the skin appendage is hair.

3. The hydrolysate is a supercritical CO hydrolysate of Hippophae rhamnoides seeds. ２ 3. The composition according to claim 1 or 2, characterized in that it is a by-product of the extraction with or a hydrolysate of a by-product obtained by pressing said seeds.

4. The composition according to any one of claims 1 to 3, characterized in that the hydrolysate is an enzymatic hydrolysate.

5. Composition according to any one of claims 1 to 4, characterized in that the hydrolysate is obtained by enzymatic digestion at alkaline pH.

6. The hydrolyzate is - a dry matter content of between 2% and 10% by weight, a total protein content of the hydrolysate between 15 g / L and 35 g / L; - a percentage of peptides with a molecular weight between 5 kDa and 30 kDa between 15% and 40%; - the percentage of peptides with a molecular weight less than 5 kDa between 40% and 85%; The composition according to any one of claims 1 to 5, characterized in that it comprises 7. A composition according to any one of claims 1 to 6, characterized in that it is applied via a topical route.

8. A composition described in any one of claims 1 to 7, characterized in that it reduces the negative effects of ageing of the skin appendages and / or of the skin and / or of the mucous membranes.

9. A composition described in any one of claims 1 to 8, characterized in that it improves the color and / or radiance and / or radiance and / or volume of skin appendages.

10. The hydrolyzate of the cake is 1×10 based on the total weight of the composition. －４ % by weight to 10% by weight, preferably 1×10 －４ Composition according to any one of claims 1 to 9, characterized in that it is contained in a content ranging from 1% to 5% by weight.

11. 11. The composition according to claim 10, characterized in that the cosmetic composition comprises at least one cosmetically acceptable additive and is selected from a serum, a lotion, a cream, a shampoo, a hair conditioner, an oil, a milk, an ointment, a paste, a foam, an emulsion, a hydrogel, a shower gel, a mask, a lacquer, a spray, a wax, a mascara, a makeup pencil or a varnish.

12. A cosmetic care composition for increasing the growth and / or reducing loss of skin appendages; and / or for maintaining and / or increasing the biomechanical and / or superficial and / or histoarchitectural properties of skin appendages; and / or for repairing damaged skin appendages, the composition comprising a hydrolysate of a cake of Hippophae rhamnoides, the composition being applied topically.

13. The composition of claim 12, wherein the skin appendage is hair.

14. A composition as described in claim 12 or 13, characterized in that it is applied topically to all or part of the skin of the body and / or face selected from the scalp, legs, thighs, arms, upper abdomen, nape, neck, part or all of the face, forehead, chin, lips, lip outline, eye outline, "T" area of ​​the face, and / or to all or part of the skin appendages.

15. A dermatological or pharmaceutical composition for its dermatological or pharmaceutical use in the treatment of alopecia and / or alopecia, the composition comprising a hydrolysate of Hippophae rhamnoides cake.

16. The composition according to claim 15, characterized in that the hydrolysate is defined in any one of claims 3 to 6.